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WO2025133391A1 - Multispecific antibodies having specificity for cd47 and pdl1 - Google Patents

Multispecific antibodies having specificity for cd47 and pdl1 Download PDF

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Publication number
WO2025133391A1
WO2025133391A1 PCT/EP2024/088374 EP2024088374W WO2025133391A1 WO 2025133391 A1 WO2025133391 A1 WO 2025133391A1 EP 2024088374 W EP2024088374 W EP 2024088374W WO 2025133391 A1 WO2025133391 A1 WO 2025133391A1
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seq
sequence
amino acid
antibody
sequences
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French (fr)
Inventor
Tea Gunde
Daniel Snell
Stefan Warmuth
Matthias Brock
Christian Hess
Philipp SCHINEIS
Marta TRÜB
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Numab Therapeutics AG
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Numab Therapeutics AG
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Priority claimed from EP23220027.9A external-priority patent/EP4574843A1/en
Priority claimed from EP23220069.1A external-priority patent/EP4574844A1/en
Application filed by Numab Therapeutics AG filed Critical Numab Therapeutics AG
Publication of WO2025133391A1 publication Critical patent/WO2025133391A1/en
Pending legal-status Critical Current
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2827—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against B7 molecules, e.g. CD80, CD86
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/55—Fab or Fab'
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance

Definitions

  • the present invention relates to a multispecific antibody comprising one binding domain, which specifically binds to CD47 (CD47-BD), one binding domain, which specifically binds to PDL1 (PDL1-BD), and optionally one or two further binding domains having specificity for an antigen different from CD47 and PDL1.
  • the present invention further relates to nucleic acid sequences encoding said multispecific antibody, vector(s) comprising said nucleic acid sequences, host cell(s) comprising said nucleic acid sequences or said vector(s), and a method of producing said multispecific antibody. Additionally, the present invention relates to pharmaceutical compositions comprising said multispecific antibody and methods of use thereof.
  • CD47 is a 50 kDa membrane receptor belonging to the immunoglobulin superfamily. CD47 consists of an extracellular N-terminal IgV domain, five transmembrane domains, and a short C-terminal intracellular tail.
  • CD47 interacts with membrane integrins, the ligand thrombospondin-1 (TSP-1) and signal-regulatory protein alpha (SIRP ⁇ ), an innate immune checkpoint protein, which is expressed on myeloid cells such as macrophages and dendritic cells (DCs) to suppress their activity.
  • TSP-1 ligand thrombospondin-1
  • SIRP ⁇ signal-regulatory protein alpha
  • CD47 is found to be overexpressed in many different tumor types (Sick E, et al., Br. J. Pharmacol., 167 (7): 1415–1430 (2012); Chao MP, et al., Curr. Opin. Immunol, 24 (2): 225–232 (2012)) and has been correlated with poor survival in patients.
  • CD47 provides a so called “don't eat me signal” to macrophages and the dendritic cells (DCs) of the immune system, enabling CD47 expressing tumor cells to escape the innate immune surveillance. [0003] Therefore, CD47 represents an interesting target in cancer therapy. However, CD47 is also ubiquitously expressed on healthy cells such as red blood cells (RBCs) and platelets in peripheral blood. Blockade of the CD47-SIRP ⁇ interaction by monoclonal antibodies has been shown to deplete those cells, often resulting in dose limiting toxicities. [0004] PDL1 (CD274, B7-H1) is a 40 kDa type I transmembrane protein.
  • PDL1 is a surface glycoprotein ligand for PD-1, a key immune checkpoint receptor expressed by activated T and B cells, and mediates immunosuppression.
  • PDL1 is found on both antigen-presenting cells and human cancer cells, such as squamous cell carcinoma of the head and neck, melanoma, and brain tumor, thyroid, thymus, esophagus, lung, breast, gastrointestinal tract, colorectum, liver, pancreas, kidney, adrenal cortex, bladder, urothelium, ovary, and skin 119915P877PC 23.12.2024 Numab Therapeutics AG (Katsuya Y, et al., Lung Cancer.88(2):154–159 (2015); Nakanishi J, et al., Cancer Immunol Immunother.56(8):1173–1182 (2007); Nomi T, et al., Clin Cancer Res.13(7):2151–2157 (2007); Fay AP, et al., J Immunother Cancer.
  • PDL1 downregulates T cell activation and cytokine secretion by binding to PD-1 (Freeman et al., 2000; Latchman et al, 2001).
  • PD-1 activated by PDL1
  • PDL1 potentially provides an immune-tolerant environment for tumor development and growth.
  • PDL1 also negatively regulates T-cell function through interaction with another receptor, B7.1 (B7-1, CD80).
  • B7.1 B7-1, CD80
  • Various antibodies against PDL1 have been developed (see, for example, WO 2013/079174 and WO 2017/118321) to be used in the treatment of PDL1 sensitive tumor types, such as those indicated above.
  • WO 2013/079174 and WO 2017/118321 to be used in the treatment of PDL1 sensitive tumor types, such as those indicated above.
  • many tumor types with low anti-PDL1 sensitivity are also enriched with CD47.
  • WO 2021/170082 discloses anti-CD47xPDL1 bispecific antibodies, wherein either the CD47 targeting moiety or the PDL1 targeting moiety is a full- length antibody, and the other targeting moiety is an antigen-binding fragment (e. g., scFv) linked directly or via a peptide linker to the N- or/and the C-terminus of the full-length antibody.
  • an antigen-binding fragment e. g., scFv
  • WO 2022/235127 discloses anti-CD47xPDL1 bispecific antibodies comprising an Fc domain, wherein the antigen-binding domain that specifically binds to CD47 is a Fab fragment located in the Fab arms, and the antigen-binding domain that specifically binds to PDL1 is an scFv fragment that is fused via a peptide linker to the C-terminus of the Fc domain.
  • WO 2019/068302 (A1) discloses anti-CD47xPDL1 bispecific antibodies that are monovalent for each specificity, wherein both binding sites are located in the Fab arms and one Fab arm is replaced by an scFv or VHH fragment.
  • WO 2021/124073 discloses IgG-based bispecific heterodimeric antibodies having one CD47-binding domain and one PDL1-binding domain that are located in the Fab arms.
  • PF- 07257876 One of these bispecific antibodies, i. e. PF- 07257876, is currently tested in phase I clinical trials.
  • WO 2019/129054 discloses IgG- like, Fc-silenced triple-chain antibodies having one CD47-binding domain located in one Fab 119915P877PC 23.12.2024 Numab Therapeutics AG arm and two VHH PDL1-binding domains that are replacing the second Fab arm.
  • One of these bispecific antibodies i. e. IBI-322, is currently tested in phase II clinical trials.
  • IBI-322 is currently tested in phase II clinical trials.
  • these anti-CD47xPDL1 bispecific antibodies have greater selectivity towards PDL1- and CD47-expressing tumor cells and/or PDL1- and CD47-expressing immune cells in a tumor microenvironment than anti-CD47 monospecific antibodies.
  • Said antibody therapeutics should have a high potency against CD47- and PDL1- expressing cancer cells and at the same time a tolerable safety profile to keep dose limiting side effects, such as in particular red blood cell agglutination, at a low level. Furthermore, it is desirable that said therapeutic antibodies have superior biophysical properties, such as a high stability, in order to facilitate developability and producibility in high yields.
  • SUMMARY OF THE INVENTION It is an object of the present invention to provide novel and improved therapeutic antibodies that can efficiently eliminate CD47- and PDL1-expressing cancer cells while exhibiting a low level of dose limiting side effects, in particular a low level of red blood cell agglutination.
  • novel and improved therapeutic antibodies exhibit high stability to facilitate developability and producibility in high yields.
  • the inventors have now surprisingly found that multispecific antibodies having one highly affine PDL1-BD and one moderately affine CD47-BD, in combination with a well- defined IgG-like antibody architecture, wherein the CD47-BD is present in the form of an scFv, exhibit efficient killing of CD47- and PDL1-expressing cancer cells, while causing virtually no agglutination or phagocytosis of red blood cells, even at higher nanomolar concentrations. This was particularly unexpected, since alternative antibody architectures, e. g.
  • CD47-BD is not in scFv format and/or the PDL1-BD and the CD47-BD are not located next to each other in the Fab arms, do not exhibit the desired combined properties, i. e. potent cancer cell lysis and at the same time low or no agglutination/lysis of red blood cells.
  • the loss of monomeric content for PRO4672, PRO5052, PRO4687 and PRO5055 which are representative multispecific antibodies of the present invention, is less than 1 % when stored for two months at 4°C at a concentration of 10 mg/ml in a 20 mM histidine buffer (pH 6.0) with 125 mM NaCl and 0.02 % Poloxamer 188.
  • This high stability is particularly surprising in view of the fact that the multispecific antibodies of the invention comprise up to four scFv antibody fragments.
  • the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i.
  • the present invention relates to a nucleic acid or two nucleic acids or three nucleic acids encoding the multispecific antibody of the present invention.
  • the present invention relates to a vector or two vectors or three vectors comprising the nucleic acid or two nucleic acids or three nucleic acids.
  • the present invention relates to a host cell or host cells comprising the vector or the two vectors or the three vectors of the present invention.
  • the present invention relates to a method for producing the multispecific antibody of the present invention, comprising (i) providing the nucleic acid or two nucleic acids or three nucleic acids of the present invention, or the vector or the two vectors or the three vectors of the present invention, expressing said nucleic acid or nucleic acids, or said vector or vectors, and collecting said multispecific antibody from the expression system, or (ii) providing a host cell or host cells of the present invention, culturing said host cell or said host cells; and collecting said multispecific antibody from the cell culture.
  • the present invention relates to a pharmaceutical composition comprising the multispecific antibody of the present invention and a pharmaceutically acceptable carrier.
  • the present invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising: a) a first multispecific antibody that specifically binds to CD47 and PDL1; b) a second antibody that specifically binds to LILRB2; and c) a pharmaceutically acceptable carrier.
  • the present invention relates to a multispecific antibody of the present invention for use as a medicament.
  • the present invention relates to a multispecific antibody of the present invention for use in the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer.
  • the present invention relates to a method for the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer, comprising the step of administering the multispecific antibody of the present invention or the pharmaceutical composition of the present invention to a patient in need thereof.
  • the present invention relates an antibody binding domain, having specificity for CD47 (CD47-BD), as defined above, wherein said CD47-BD is in a format selected from an Fv, a dsFv, an scFv and a dsscFv, particularly from an Fv and an scFv, particularly wherein said CD47-BD is in the form of an scFv.
  • a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD), wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequence of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequence of SEQ ID NO: 7; or ii.
  • the multispecific antibody of item 1 wherein said CD47-BD comprises: the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequence of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequence of SEQ ID NO: 7.
  • said PDL1-BD comprises: i.
  • the HCDR1 sequence of SEQ ID NO: 20 the HCDR2 sequence of SEQ ID NO: 21, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequence of SEQ ID NO: 27; or v. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequence of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequence of SEQ ID NO: 27; or vi.
  • KD monovalent dissociation constant
  • CD47-BD when in scFv format, has a melting temperature (Tm), determined by differential scanning fluorimetry, in the range of 50°C to 70°C, particularly in the range of 55°C to 65°C, in particular wherein said antibody is formulated in 50 mM phosphate- citrate buffer (NaCiP) at pH 6.4, 150 mM NaCl; b.
  • Tm melting temperature
  • the antibody of the invention when in scFv format, has a loss in monomer content, after storage for four weeks at ⁇ 80°C, of less than 3 %, less than 2 %, less than 1 %, particularly less than 0.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer (NaCiP) with 150 mM NaCl at pH 6.4; c.
  • NaCiP phosphate citrate buffer
  • the antibody of the invention when in scFv format, has a loss in monomer content, after storage for four weeks at 4°C, of less than 3 %, less than 2 %, less than 1 %, particularly less than 0.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer (NaCiP) with 150 mM NaCl at pH 6.4; and/or d.
  • NaCiP phosphate citrate buffer
  • Tm melting temperature
  • the antibody of the invention when in scFv format, has a loss in monomer content, after five consecutive freeze- thaw cycles, of less than 5 %, preferably less than 3 %, more preferably less than 1 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein the antibody of the invention is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4; and/or c. when in scFv format, has a loss in monomer content, after storage for four weeks at 4°C, of less than 15 %, e. g.
  • the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4. 9.
  • the multispecific antibody of any of the preceding items wherein said multispecific antibody does not trigger any visible red blood cell agglutination at a concentration of 5 nM, particularly of 20 nM, particularly of 50 nM, particularly of 100 nM, particularly of 200 nM, particularly of 500 nM, as determined in a standard hemagglutination assay as 119915P877PC 23.12.2024 Numab Therapeutics AG described in Example 4.2; and/or does not cause the lysis of red blood cells at a concentration of 5 nM, particularly of 20 nM, particularly of 50 nM, as determined in a standard phagocytosis assay, as described in Example 5.4. 11.
  • the multispecific antibody of item 1 wherein the immunoglobulin Fc region is selected from an immunoglobulin Fc region of an IgG subclass, particularly an immunoglobulin Fc region of human lgG1, lgG2 (lgG2A, IgG2B), lgG3 or lgG4, particularly of human IgG1 or IgG2, particularly of human IgG1. 12.
  • the multispecific antibody of item 1, wherein the immunoglobulin Fc region is an active Fc region. 13.
  • the multispecific antibody of any one of the preceding items wherein the immunoglobulin Fc region has one or more amino acid modifications selected from the group consisting of: a) modifications to enhance heterodimerization of heavy chains, such as the knob- into-hole (KiH) technology; b) modifications that increase or decrease Fc receptor-binding, when compared to the respective unmodified immunoglobulin Fc region thereof; c) modifications that increase serum half-life, when compared to the respective unmodified immunoglobulin Fc region thereof. 14.
  • said CD47-BD comprises VH3 or VH4 domain framework sequences FR1 to FR4; particularly VH3 domain framework sequences FR1 to FR4. 15.
  • CD47-BD comprises a) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 8 and 9; and 119915P877PC 23.12.2024 Numab Therapeutics AG a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 11 and 12, particularly to SEQ ID NO: 12; or b) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 10; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 10; and a VL sequence
  • CD47-BD comprises a) a VH sequence selected from any one of SEQ ID NOs: 8 and 9; and a VL sequence selected from any one of SEQ ID NOs: 11 and 12, particularly the VL sequence of SEQ ID NO: 12; or b) the VH sequence of SEQ ID NO: 10; and the VL sequence of SEQ ID NO: 13.
  • PDL1-BD comprises VH3 or VH4 domain framework sequences FR1 to FR4; particularly VH3 domain framework sequences FR1 to FR4. 19.
  • said PDL1-BD comprises a) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 28; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 34, 36 and 37, particularly selected from SEQ ID NOs: 34 and 37, particularly to SEQ ID NO:37; or b) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 29 and 31; and 119915P877PC 23.12.2024 Numab Therapeutics AG a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to
  • the multispecific antibody of any one of the preceding items, wherein said PDL1-BD comprises a) the VH sequence of SEQ ID NO: 28; and a VL sequence selected from any one of SEQ ID NOs: 34, 36 and 37, particularly selected from any one of SEQ ID NOs: 34 and 37, particularly the VL sequence of SEQ ID NO: 37; or b) a VH sequence selected from any one of SEQ ID NOs: 29 and 31; and a VL sequence selected from any one of SEQ ID NOs: 33 and 35, particularly the VL sequence of SEQ ID NO: 35; or c) the VH sequence of SEQ ID NO: 30; and a VL sequence selected from any one of SEQ ID NOs: 32 and 34, particularly the VL sequence of SEQ ID NO: 34; or d) a VH sequence selected from any one of SEQ ID NOs: 38 and 42; and a VL sequence selected from any one of SEQ ID NOs: 48, 50 and 51, particularly from SEQ ID NOs: 48
  • the multispecific antibody of any one of the preceding items, wherein said PDL1-BDs comprises CDR regions derived from a parental rabbit antibody.
  • said CD47-BD comprises: a) a VH sequence selected from any one of SEQ ID NOs: 8 and 9; and the VL sequence of SEQ ID NO: 12; or b) the VH sequence of SEQ ID NO: 10; and the VL sequence of SEQ ID NO: 13; and wherein said PDL1-BD comprises: a) the VH sequence of SEQ ID NO: 52; and a VL sequence selected from any one of SEQ ID NOs: 58 and 61, particularly the VL sequence of SEQ ID NO:61; or b) the VH sequence of SEQ ID NO: 54; and the VL sequence of SEQ ID NO: 58; or d) the VH sequence of SEQ ID NO: 81; and a VL sequence selected from any one of SEQ ID NOs: 86 and
  • the multispecific antibody of any one of the preceding items which has amino acid sequences selected from: - SEQ ID NO: 174 and SEQ ID NO: 175; - SEQ ID NO: 176 SEQ ID NO: 177 and SEQ ID NO: 178; - SEQ ID NO: 179, SEQ ID NO: 180 and SEQ ID NO: 181; - SEQ ID NO: 182 and SEQ ID NO: 183; - SEQ ID NO: 184, SEQ ID NO: 185 and SEQ ID NO: 186; 119915P877PC 23.12.2024 Numab Therapeutics AG - SEQ ID NO: 197 and SEQ ID NO: 198; - SEQ ID NO: 199, SEQ ID NO: 200 and SEQ ID NO: 201; - SEQ ID NO: 202 and SEQ ID NO: 203; and - SEQ ID NO: 204, SEQ ID NO: 205 and SEQ ID NO: 206.
  • the multispecific antibody of any one of the preceding items wherein said multispecific antibody further comprises: d) one further binding domain, which has specificity for an antigen different from CD47 and PDL1, wherein said one further binding domain is attached to one of the C-termini of the heavy chains of the immunoglobulin Fc region.
  • 26. The multispecific antibody of any one of the items 1 to 24, wherein the multispecific antibody further comprises: d) two further binding domains, which have specificity for an antigen different from CD47 and PDL1, wherein said two further binding domains are attached to each of the C-termini of the heavy chains of the immunoglobulin Fc region.
  • TAA tumor associated antigen
  • the TAA is selected from ADRB3, AFP, ALK, BCMA, beta human chorionic gonadotropin, CA-125 (MUC16), CAIX, CD123, CD133, CD135, CD135 (FLT3), CD138, CD171, CD19, CD20, CD22, CD24, CD276, CD33, CD33, CD38, CD44v6, CD79b, CD97, CDH3 (cadherin 3), CEA, CEACAM6, CLDN6, CLEC12A (CLL1), CSPG4, CYP1B1, EGFR, EGFRvlll, EPCAM, EPHA2, Ephrin B2, ERBBs (e.
  • ERBB2 g. ERBB2
  • FAP FAP
  • FGFR1 folate receptor alpha
  • folate receptor beta Fos-related antigen
  • GA733, GD2, GD3, GFRalpha4 globoH
  • GPC3, GPR20 GPRC5D
  • HAVCR1 Her2/neu (HER2)
  • HLA-A2, HMWMAA HPV E6 or E7
  • human telomerase reverse transcriptase IL-11Ra, IL-13Ra2, intestinal carboxyl esterase, KIT, Legumain, LewisY, LMP2, Ly6k
  • MAD-CT-1 MAD-CT-2
  • ML-IAP MN-CA IX
  • MSLN MUC1, mut hsp 70-2, NA-17, NCAM, neutrophil elastase, NY-BR-1, NY-ESO- 1, o-acetyl-GD2, OR51E2, PANX3, PDGFR-beta, PLAC1, Polysialic acid,
  • the multispecific antibody of item 29 or 30, wherein the immunomodulatory antigen different from PDL1 is selected from B7-1 (CD80), B7-2 (CD86), B7-DC (CD273), B7-H1 (CD274), B7-H2 (CD275), B7-H3 (CD276), B7-H4 (VTCN1), B7-H5 (VISTA), BTLA (CD272), 4-1BB (CD137), CD137L, CD24, CD27, CD28, CD38, CD40, CD40L (CD154), CD59, CD70, CTLA-1, CTLA-4 (CD152), CXCL9, GITR (CD357), GITR, HVEM (CD270), ICAM-1 (CD54), ICOS (CD278), LAG-3 (CD223), OX40 (CD134), OX40L (CD252), PD-1 (CD279), TIGIT, CD314, CD334, CD335, CD337, VISTA and TIM-3 (CD36
  • NK cell receptor is selected from KIR2DL1, KIR2DL2/3, KIR2DL4, KIR2DL5A, KIR2DL5B, KIR3DL1, KIR3DL2, KIR3DL3, KIR2DS4, KIR3DS, NKG2A, NKG2C, NKG2D, NKG2E, NKp30, NKp44, NKp46, CD2, CD16, CD69, DNAX accessory molecule-1 (DNAM-1), 2B4, NK1.1; a killer immunoglobulin (Ig)-like activating receptors (KAR); ILTs/LIRs (CD85) such as LILRA1, LILRA2, LILRA3, LILRA4, LILRA5, LILRA6, LILRB1, LILRB2, LILRB3, LILRB4, LILRB5, ILT8, ILT10,
  • NK cell receptor is selected from NKp46, LILRB1, LILRB2, LILRB3, LILRB4 and LILRB5, particularly LILRB1, LILRB2, and LILRB4, particularly LILRB1 and LILRB2, particularly LILRB2.
  • NK cell receptor is selected from NKp46, LILRB1, LILRB2, LILRB3, LILRB4 and LILRB5, particularly LILRB1, LILRB2, and LILRB4, particularly LILRB1 and LILRB2, particularly LILRB2.
  • said further binding domain specifically binds to LILRB2 (LILRB2-BD).
  • said two further binding domains specifically bind to LILRB2 (LILRB2-BDs). 119915P877PC 23.12.2024 Numab Therapeutics AG 37.
  • the multispecific antibody of item 36, wherein the LILRB2-BDs bind to different epitopes on the extracellular domain of LILRB2. 38.
  • the multispecific antibody of item 36, wherein the LILRB2-BDs bind to the same epitope on the extracellular domain of LILRB2. 39.
  • the multispecific antibody of item 36, wherein the LILRB2-BDs are identical.
  • 40. The multispecific antibody of any one of the items 35 to 39, wherein said LILRB2-BD or said LILRB2-BDs comprise: i.
  • the HCDR1 sequence of SEQ ID NO: 112 the HCDR2 sequence of SEQ ID NO: 113, the HCDR3 sequence of SEQ ID NO: 114, the LCDR1 sequence of SEQ ID NO: 115, the LCDR2 sequence of SEQ ID NO: 117, and the LCDR3 sequence of SEQ ID NO: 118; or iv. the HCDR1 sequence of SEQ ID NO: 112, the HCDR2 sequence of SEQ ID NO: 113, the HCDR3 sequence of SEQ ID NO: 114, the LCDR1 sequence of SEQ ID NO: 115, the LCDR2 sequence of SEQ ID NO: 116, and the LCDR3 sequence of SEQ ID NO: 118 or v.
  • KD monovalent dissociation constant
  • SPR surface plasmon resonance
  • LILRB2 block the interaction of LILRB2 to its ligand HLA-A3 with an IC50 of 100 pM to 10 nM, particularly of 100 pM to 5 nM, particularly of 100 pM to 2 nM, as measured by flow cytometry; d. are not cross-reactive with members of leukocyte immunoglobulin-like receptor subfamily A (LILRA); and/or e. are not cross reactive with members of leukocyte immunoglobulin-like receptor subfamily LILRB3, LILRB4 and LILRB5.
  • LILRA leukocyte immunoglobulin-like receptor subfamily A
  • LILRB3 leukocyte immunoglobulin-like receptor subfamily LILRB3, LILRB4 and LILRB5.
  • Tm melting temperature
  • LILRB2-BD or said LILRB2-BDs comprise VH3 or VH4 domain framework sequences FR1 to FR4; particularly VH3 domain framework sequences FR1 to FR4. 48.
  • LILRB2-BD or said LILRB2-BDs independently comprise a) a VL domain comprising a VL framework comprising framework regions FR1, FR2, FR3 and FR4, which are selected from V ⁇ subtypes, particularly from the V ⁇ 1 and V ⁇ 3 subtypes, particularly are of the V ⁇ 1 subtype; or b) a VL domain comprising a VL framework comprising framework regions FR1, FR2 and FR3, which are selected from V ⁇ subtypes, particularly from the V ⁇ 1 and V ⁇ 3 subtypes, particularly are of the V ⁇ 1 subtype, and a framework FR4, which is selected from a V ⁇ FR4, particularly is a V ⁇ FR4 comprising an amino acid sequence having at least 80, 90 percent identity to any of SEQ ID NOs: 161 to 169, more particularly a V ⁇ FR4 selected from any of SEQ ID NOs: 161 to 169, particularly a V ⁇ FR4 according to SEQ
  • LILRB2-BD or said LILRB2-BDs independently comprise a) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 100; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 104, 106 and 107, particularly selected from SEQ ID NOs: 104 and 107, particularly to SEQ ID NO: 107; or b) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 101; and a VL sequence at least 90, 91,
  • LILRB2-BD or said LILRB2-BDs independently comprise a) a VH sequence of SEQ ID NO: 100; and 119915P877PC 23.12.2024 Numab Therapeutics AG a VL sequence selected from SEQ ID NOs: 104, 106 and 107, particularly selected from SEQ ID NOs: 104 and 107, particularly from SEQ ID NO: 107; or b) a VH sequence of SEQ ID NO: 101; and a VL sequence selected from SEQ ID NOs: 103 and 104, particularly from SEQ ID NO: 104; or c) a VH sequence of SEQ ID NO: 102; and a VL sequence of SEQ ID NO: 105; or d) a VH sequence of SEQ ID NO: 119; and a VL sequence selected from SEQ ID NOs: 122, 123 and 124, particularly selected from SEQ ID NOs: 122 and 124, particularly from SEQ ID NO: 122 and 124, particularly from SEQ ID NO:
  • the antibody of the invention when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 20 mM Histidine buffer at pH 6.0, 125 mM NaCl and 0.02 % Poloxamer 188; and/or k) has a loss in monomer content, after storage for 4 weeks at 40°C, of less than 7 %, e. g.
  • the multispecific antibody of any one of the items 35 to 52 which has amino acid sequences selected from: - SEQ ID NO: 187 and SEQ ID NO: 188 (PRO4672); - SEQ ID NO: 189, SEQ ID NO: 190 and SEQ ID NO: 191 (PRO4687); - SEQ ID NO: 192 and SEQ ID NO: 193 (PRO5052); and - SEQ ID NO: 194, SEQ ID NO: 195 and SEQ ID NO: 196 (PRO5055).
  • a vector or two vectors or three vectors comprising the nucleic acid or two nucleic acids or three nucleic acids of item 54.
  • 56. A host cell or host cells comprising the vector or the two vectors or the three vectors of item 55.
  • a method for producing the multispecific antibody of any one of items 1 to 53 comprising (i) providing the nucleic acid or two nucleic acids or three nucleic acids of item 54, or the 119915P877PC 23.12.2024 Numab Therapeutics AG vector or the two vectors or the three vectors of item 55, expressing said nucleic acid or nucleic acids, or said vector or vectors, and collecting said multispecific antibody from the expression system, or (ii) providing a host cell or host cells according to item 56, culturing said host cell or said host cells; and collecting said multispecific antibody from the cell culture.
  • a pharmaceutical composition comprising the multispecific antibody of any one of items 1 to 53 and a pharmaceutically acceptable carrier.
  • a pharmaceutical composition comprising: a) a first multispecific antibody that specifically binds to CD47 and PDL1; b) a second antibody that specifically binds to LILRB2; and c) a pharmaceutically acceptable carrier. 60.
  • the pharmaceutical composition of item 59 wherein the first antibody that specifically binds to CD47 and PDL1 is selected from anti-CD47xPDL1 antibodies that are currently tested or have been tested in clinical trials, preferably wherein the first antibody that specifically binds to CD47 and PDL1 is selected from the group consisting of: IBI-322 by Innovent Biologics Inc.; PF-07257876 by Pfizer Inc.; 6MW-3211 by Mabwell (Shanghai) Bioscience Co.
  • the pharmaceutical composition of any one of the items 59 to 61, wherein the second antibody that specifically binds to LILRB2 comprises at least one LILRB2-BD as defined in any one of the items 40 to 50.
  • the pharmaceutical composition of any one of the items 59 to 61 and 63, wherein the second antibody comprises two LILRB2-BDs.
  • the pharmaceutical composition of item 64, wherein the two LILRB2-BDs bind to different epitopes on the extracellular domain of LILRB2.
  • the pharmaceutical composition of item 64, wherein the two LILRB2-BDs bind to the same epitope on the extracellular domain of LILRB2.
  • the pharmaceutical composition of item 66, wherein the LILRB2-BDs are identical. 68.
  • the format of the second antibody is selected from a tandem scDb (Tandab), a linear dimeric scDb (LD-scDb), a circular dimeric scDb (CD-scDb), a tandem tri-scFv, a tribody (Fab-(scFv)2), a Fab-Fv2, a triabody, an scDb-scFv, a tetrabody, a didiabody, a tandem-di-scFv and a MATCH.
  • the multispecific antibody of item 74, wherein the cancer is selected from a benign or especially malignant tumor, solid tumors, mesothelioma, brain cancer, kidney cancer, liver cancer, adrenal gland cancer, bladder cancer, breast cancer, stomach cancer (e.
  • gastric tumors e.g., gastric tumors
  • esophageal cancer ovarian cancer, cervical cancer, thymic cancer, choriocarcinoma, oral cancer, salivary cancer, colon cancer, rectum cancer, prostate cancer, pancreatic cancer, lung cancer (e. g., non-small cell lung cancer and small cell lung cancer), vaginal cancer, thyroid cancer, melanoma (e.
  • melanoma g., unresectable or metastatic melanoma
  • renal cell carcinoma sarcoma
  • glioma glioblastoma
  • multiple myeloma or gastrointestinal cancer especially colon carcinoma or colorectal adenoma
  • a tumor of the neck and head endometrial cancer
  • Cowden syndrome Lhermitte-Duclos disease
  • Bannayan-Zonana syndrome prostate hyperplasia
  • a neoplasia especially of epithelial character, preferably mammary carcinoma or squamous cell carcinoma, chronic lymphocytic leukemia, chronic myelogenous leukemia (e.
  • the method of item 76, wherein the cancer is selected from 119915P877PC 23.12.2024 Numab Therapeutics AG a benign or especially malignant tumor, solid tumors, mesothelioma, brain cancer, kidney cancer, liver cancer, adrenal gland cancer, bladder cancer, breast cancer, stomach cancer (e. g., gastric tumors), esophageal cancer, ovarian cancer, cervical cancer, thymic cancer, choriocarcinoma, oral cancer, salivary cancer, colon cancer, rectum cancer, prostate cancer, pancreatic cancer, lung cancer (e. g., non-small cell lung cancer and small cell lung cancer), vaginal cancer, thyroid cancer, melanoma (e.
  • a benign or especially malignant tumor solid tumors, mesothelioma, brain cancer, kidney cancer, liver cancer, adrenal gland cancer, bladder cancer, breast cancer, stomach cancer (e. g., gastric tumors), esophageal cancer, ovarian cancer, cervical cancer, thymic
  • melanoma g., unresectable or metastatic melanoma
  • renal cell carcinoma sarcoma
  • glioma glioblastoma
  • multiple myeloma or gastrointestinal cancer especially colon carcinoma or colorectal adenoma
  • a tumor of the neck and head endometrial cancer
  • Cowden syndrome Lhermitte-Duclos disease
  • Bannayan-Zonana syndrome prostate hyperplasia
  • a neoplasia especially of epithelial character, preferably mammary carcinoma or squamous cell carcinoma, chronic lymphocytic leukemia, chronic myelogenous leukemia (e.
  • An antibody comprising one or two, preferably one, antibody binding domains according to item 78 or 79.
  • Figure 2 shows the binding properties of PRO3635 (IgG of scFv PRO3064, 68-18- E11-sc02) and the references PRO3483 (NGM), PRO2518 (J19) and PRO3848 (Biond) to members of LILRA family expressed by CHO cells.
  • An anti-V5 tag antibody served as positive control as all constructs contain V5 tag at N-terminus.
  • Figure 3 shows the binding properties of PRO3635 (IgG of scFv PRO3064, 68-18- E11-sc02) and the references PRO3483 (NGM), PRO2518 (J19) and PRO3848 (Biond) to members of LILRB family expressed by CHO cells.
  • FIG. 4 shows the binding of the scFv PRO3153 (69-01-E02-sc03) applied at concentrations of 150 nM (A) and 50 nM (B) to members of LILRBA and LILRB family expressed by CHO cells.
  • Figure 5 shows the binding properties of the scFv PRO3697 (69-06-G04-sc05), cross-linked with an anti-framework antibody (x-linked), to members of LILRBA and LILRB family expressed by CHO cells.
  • Figure 6 shows the dose-dependent increase of the M1 marker CD86 and the dose-dependent decrease of the M2 marker CD163 in the presence of increasing concentrations of PRO3744, PRO3696 and PRO3697.
  • Figure 10 shows the phagocytosis potency of trispecific antibodies PRO4672, PRO4687, PRO5052 and PRO5055 and of the reference compounds PRO2777, PRO3599 and PRO3851 for H292 cancer cells (A) and (C), expressing PDL1 and CD47, and for NALM6 cancer cells (B) and A375 cancer cells (D), expressing only CD47.
  • Figure 11 shows the phagocytosis potency of trispecific antibodies PRO5052 and PRO5055 and of the reference compounds PRO2777, avelumab, PRO2777 + avelumab combination, PRO5134 (silenced Fc), PRO5054 (no CD47-BD) and PRO5057 (no CD47- BD) for H292 cancer cells (A) and (C), expressing PDL1 and CD47, and for A375 cancer cells (B) and (D), expressing only CD47.
  • Figure 12 shows the phagocytosis potency of trispecific antibodies PRO5052, PRO5053, PRO5055 and PRO5056 and of the reference compounds PRO2777, PRO2777 + PRO2734 combination, PRO2777 + PRO2734 + avelumab combination, PRO5134 (silenced Fc), PRO5054 (no CD47-BD) and PRO5057 (no CD47-BD) for H292 cancer cells (A) and (C), expressing PDL1 and CD47, and for A375 cancer cells (B) and (D), expressing only CD47.
  • Figure 13 shows the phagocytosis potency of the trispecific antibodies PRO4672, PRO4687, PRO5055, of the bispecific antibody PRO3790, of the comparative antibodies PRO4682 (CD47-BD in Fab), PRO4711 (CD47-BD from PRO3599), PRO4677 (inverse Morrison) and of the reference compounds PRO2777, PRO3599, for H292 cancer cells (A and C) and red blood cells (RBCs) (B C and D).
  • Figure 14 shows the phagocytosis potency of the trispecific antibodies PRO4672, PRO4673, PRO4687, PRO4688, of the comparative antibodies PRO4682 and PRO4683 (CD47-BD in Fab), and of the reference antibodies PRO2777, PRO3609 and PRO3613, for H292 cancer cells (A and C) and red blood cells (RBCs) (B and D).
  • Figure 15 shows the phagocytosis potency of the multispecific antibodies PRO5052, PRO5053, PRO5055 and PRO5056 towards H292 tumor cell and red blood cells (RBCs) in parallel to illustrates the potential therapeutic window.
  • Figure 16 shows the T-cell proliferation potency of the anti-LILRB2 antibody PRO4710, the trispecific antibodies PRO5052, PRO5053 (no-LILRB2-BD), PRO5055, PRO5056 (no-LILRB2-BD), PRO5054 (no CD47-BD), and the reference antibodies avelumab and magrolimab.
  • Figure 17 shows the release of TNF ⁇ and IL-1 ⁇ mediated by the anti-LILRB2 antibody PRO4710, the trispecific antibodies PRO4672, PRO4673 (no LILRB2-BD), PRO4687, PRO4688 (no LILRB-BD), PRO5052, PRO5055, PRO5056 (no LILRB2-BD), by the comparative antibodies PRO5054 (no CD47-BD), PRO3613 (medium affinity CD47-BD), and by the reference antibodies magrolimab, atezolizumab (Atezo), PRO3599, PRO3851 and PRO2777.
  • Figure 18 shows the hemagglutination assay result for trispecific antibodies PRO5052, PRO5055, and reference antibodies PRO2777, PRO3599.
  • Figure 19 shows the hemagglutination assay result for the trispecific antibodies PRO5052, PRO5055, PRO4672, PRO4687, and reference antibodies PRO2777, PRO3851 and PRO3599.
  • Figure 20 shows the binding of PRO5052, PRO5055 to cancer cell lines HT-29 (A), the binding of PRO5052 and PRO4672 to cancer cell line HT-1080 (B), the binding of PRO4687, PRO5055 and reference antibodies PRO3851 and PRO2777 to cancer cell lines HT-29 (C) and HT-1080 (D).
  • PRO2777 and PRO3851 were applied as reference antibodies.
  • Figure 21 shows the inhibition of CD47/SIRP ⁇ interaction on CHO-CD47 cells (A), NCI-H292 cells (B) and NCI-H292 cells treated with 10 ng/ml INFy (C) for PRO4672, PRO5052 and the reference antibodies PRO2777 and PRO3599.
  • Figure 22 shows the inhibition of CD47/SIRP ⁇ interaction on CHO-CD47 cells (A), NCI-H292 cells (B), NCI-H292 cells treated with 10 ng/ml INFy (C) for PRO4687, PRO5055 and the reference antibodies PRO2777 and PRO3851.
  • Figure 23 shows the binding properties of PRO5052 (A) and PRO5055 (B) to members of LILRBA and LILRB family expressed by CHO cells.
  • Figure 24 shows the formats of the multispecific antibodies PRO3790, PRO5052, PRO5053, PRO5055, PRO5056 and the comparative antibodies PRO4677, PRO4682 and PRO4683.
  • the present invention provides multispecific antibodies comprising one CD47-BD and one PDL1-BD, and an IgG-like region comprising an immunoglobulin Fc region, a first Fab arm and a second Fab arm, wherein the CD47 BD is an scFv fragment replacing the first Fab arm, and the PDL1 BD is located in the second Fab arm or is an scFv fragment replacing said second Fab arm.
  • These multispecific antibodies do not cause phagocytosis of red blood cells or trigger any visible red blood cell agglutination even at high nanomolar 119915P877PC 23.12.2024 Numab Therapeutics AG concentrations.
  • antibody formats wherein the CD47-BD is in Fab format exhibit significant red blood cell agglutination.
  • Fc silenced IgG-like formats or Fc less antibody formats do not provide the desired cancer cell killing potency.
  • PRO3790, PRO4672, PRO4673, PRO4687, PRO4688, PRO5052, PRO5053, PRO5055, PRO5056 exhibit lower red blood cell agglutination and/or red blood cell lysis than the prior art anti-CD47xPDL1 antibodies PF-07257876 (Pfizer Inc.) and IBI-322 (Innovent Biologics Inc), while exhibiting similar or even better in-vitro cancer cell killing potency. It is thus expected that the multispecific antibodies of the present invention will provide a large therapeutic window and thus provide distinct therapeutic advantages over conventional therapies. [0056] In addition, the multispecific antibodies of the present invention exhibit very advantageous biophysical properties, in particular high storage stability.
  • the loss of monomeric content for PRO4672, PRO5052, PRO4687 and PRO5055 which are representative multispecific antibodies of the present invention, is less than 1 % when stored for two months at 4°C at a concentration of 10 mg/ml in a 20 mM Histidine buffer (pH 6.0) with 125 mM NaCl and 0.02 % Poloxamer 188.
  • This high stability is particularly surprising in view of the fact that the multispecific antibodies of the invention comprise up to four scFv antibody fragments.
  • antibody and the like, as used herein, includes whole antibodies or single chains thereof; and any antigen-binding fragment (i. e., “antigen-binding portion”) or single chains thereof; and molecules comprising antibody CDRs, VH regions or VL regions (including without limitation multispecific antibodies).
  • a naturally occurring “whole antibody” is a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter- connected by disulfide bonds. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region.
  • the heavy chain constant region is comprised of three domains, CH1, CH2 and CH3.
  • Each light chain is comprised of a light chain variable region (abbreviated herein as VL) and a light chain constant region.
  • the light chain constant region is comprised of one domain, CL.
  • the VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), flanked by regions that are more conserved, termed framework regions (FRs).
  • CDRs complementarity determining regions
  • FRs framework regions
  • Each VH and VL is composed of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
  • variable regions of the heavy and light chains contain a binding domain that interacts with an antigen.
  • the constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e. g., effector cells) and the first component (C1q) of the classical complement system.
  • the term “immunoglobulin Fc region” or “Fc region”, as used herein, is used to define a C-terminal region of an immunoglobulin heavy chain, i. e. the CH2 and CH3 domains of the heavy chain constant regions.
  • the Fc region provides linkage to humoral and cellular components of the immune system, mostly through interaction with C1q and Fc receptors (FcR).
  • Fc region includes Fc regions of an IgG subclass, particularly an immunoglobulin Fc region of human lgG1, lgG2 (lgG2A, IgG2B), lgG3 or lgG4. Further included are Fc regions that are engineered, i. e. Fc regions that have one or more amino acid modifications, to exhibit certain desired properties.
  • amino acid modifications are for example modifications to enhance heterodimerization of heavy chains, such as knob-into- 119915P877PC 23.12.2024 Numab Therapeutics AG hole (KiH) technology (see for example Ridgway et al., Protein Eng.9:617-21 (1996) and Spiess et al., J Biol Chem.288(37):26583-93 (2013)).
  • Other amino acid modifications are modifications that alter, i. e.
  • Fc receptor or “FcR” describes a receptor that binds to the Fc region of an antibody.
  • the Fc receptors are native sequence human FcRs, which binds an IgG antibody. These include for example the Fc gamma receptors (Fc ⁇ Rs).
  • activating Fc ⁇ Rs Five activating Fc ⁇ Rs are known, i. e. the high affinity receptors Fc ⁇ RI, and the lower affinity receptors Fc ⁇ RIIA, Fc ⁇ RIIC, Fc ⁇ RIIIA and Fc ⁇ RIIIB.
  • one inhibiting receptor is known, i. e. Fc ⁇ RIIB, which have similar amino acid sequences to Fc ⁇ RIIA that differ primarily in the cytoplasmic domains thereof.
  • Activating receptors contain an immunoreceptor tyrosine- based activation motif (ITAM) in its cytoplasmic domain.
  • the inhibiting receptor Fc ⁇ RIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain, (see M.
  • FcRs are reviewed for example in Ravetch and Kinet, Annu. Rev. Immunol.9: 457-92 (1991); Capet et al, Immunomethods 4: 25-34 (1994); and de Haas et al, J. Lab. Clin. Med.126: 330-41 (1995).
  • Other FcRs including those to be identified in the future, are encompassed by the term “FcR” herein.
  • the term “Fc receptor” or “FcR” also includes the neonatal receptor FcRn, which plays a central role in the cellular trafficking and serum half-life of IgGs.
  • siRNAs have been engineered to exhibit insignificant or negligible Fc receptor- binding functionality, in particular insignificant or negligible binding affinity to Fc ⁇ Rs, when compared to its unmodified version.
  • Antibodies having such silenced Fc regions do not or do not significantly trigger cellular immune responses through Fc receptor-binding.
  • active Fc or “active Fc region”, as used herein, refers to an Fc region, which has significant Fc receptor-binding functionality, in particular significant binding affinity to Fc ⁇ Rs. This Fc receptor-binding functionality may be present naturally and/or may be introduced or enhanced by engineering. Antibodies having such active Fc regions trigger cellular immune responses through Fc receptor-binding.
  • the immunoglobulin Fc region comprised in the multispecific antibodies of the invention is selected from an Fc region of an IgG subclass, 119915P877PC 23.12.2024 Numab Therapeutics AG particularly an immunoglobulin Fc region of human lgG1, lgG2 (lgG2A, IgG2B), lgG3 or lgG4, particularly of human IgG1 or IgG2, particularly of human IgG1.
  • the immunoglobulin Fc region comprised in the multispecific antibodies of the invention is an active Fc region.
  • the immunoglobulin Fc region comprised in the multispecific antibodies of the invention has one or more amino acid modifications selected from the group consisting of: a) modifications to enhance heterodimerization of heavy chains, such as the knob-into-hole (KiH) technology; b) modifications that increase or decrease, in particular increase, Fc receptor-binding, when compared to the respective unmodified immunoglobulin Fc region thereof; c) modifications that increase serum half-life, when compared to the respective unmodified immunoglobulin Fc region thereof.
  • a) modifications to enhance heterodimerization of heavy chains such as the knob-into-hole (KiH) technology
  • modifications that increase or decrease, in particular increase, Fc receptor-binding when compared to the respective unmodified immunoglobulin Fc region thereof
  • modifications that increase serum half-life when compared to the respective unmodified immunoglobulin Fc region thereof.
  • the immunoglobulin Fc region comprised in the multispecific antibodies of the invention has one or more amino acid modifications to enhance heterodimerization of the heavy chains, such as the knob-into-hole (KiH) technology.
  • binding domain refers to one or more fragments of an intact antibody that retain the ability to specifically bind to a given antigen (e. g., CD47, PDL1, LILRB2).
  • Antigen-binding functions of an antibody can be performed by fragments of an intact antibody.
  • binding domain refers to a Fab fragment, i. e. a monovalent fragment consisting of the VL, VH, CL and CH1 domains; a single chain Fab, i. e. a Fab fragment, where the heavy chain and the light chain are connected via a linker; an Fv fragment consisting of the VL and VH domains of a single arm of an antibody; a disulfide stabilized Fv fragment (dsFv); and a single chain Fv fragment (scFv).
  • Fab fragment i. e. a monovalent fragment consisting of the VL, VH, CL and CH1 domains
  • single chain Fab i. e. a Fab fragment, where the heavy chain and the light chain are connected via a linker
  • an Fv fragment consisting of the VL and VH domains of a single arm of an antibody
  • dsFv disulfide stabilized Fv fragment
  • scFv single chain Fv fragment
  • the binding domains of the antibodies of the present invention are independently of each other selected from a Fab fragment, an Fv fragment, a dsFv fragment and a single-chain Fv fragment (scFv).
  • the binding domains of the antibodies of the present invention are independently of each other selected from a Fab fragment and a single-chain Fv fragment (scFv).
  • the VL and VH domains of the scFv fragment are stabilized by an interdomain disulfide bond (dsscFv), in particular said VH domain comprises a single cysteine residue in position 51 (AHo numbering) and said VL domain comprises a single cysteine residue in position 141 (AHo numbering).
  • fragment-based refers to an antibody that comprises at least one antigen-binding fragment, as defined above, but no immunoglobulin Fc region. Due to the lack of the Fc region, these antibodies typically comprise and hSA-binding domain for half-life extension.
  • CDRs Complementarity Determining Regions
  • the CDR amino acid residues in the heavy chain variable region (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable region (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3).
  • the CDR amino acids in the VH are numbered 26-32 (HCDR1), 52-56 (HCDR2), and 95-102 (HCDR3); and the amino acid residues in VL are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3).
  • the CDRs consist of amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in human VH and amino acid residues 24-34 (LCDR1), 50- 56 (LCDR2), and 89-97 (LCDR3) in human VL.
  • HCDR1 amino acid residues 26-35
  • LCDR2 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • LCDR3 amino acid residues 24-34
  • the CDRs of an antibody can be determined using the program IMGT/DomainGap Align.
  • IMGT/DomainGap Align the program IMGT/DomainGap Align.
  • AHo Honegger & Plückthun
  • CDRs according to AHo numbering scheme: LCDR1 (also referred to as CDR- L1): L24-L42; LCDR2 (also referred to as CDR-L2): L58-L72; LCDR3 (also referred to as CDR-L3): L107-L138; HCDR1 (also referred to as CDR-H1): H27-H42; HCDR2 (also referred to as CDR-H2): H57-H76; HCDR3 (also referred to as CDR-H3): H108-H138.
  • LCDR1 also referred to as CDR- L1
  • LCDR2 also referred to as CDR-L2
  • LCDR3 also referred to as CDR-L3
  • the numbering system according to Honegger & Plückthun takes the length diversity into account that is found in naturally occurring antibodies, both in the different VH and VL subfamilies and, in particular, in the CDRs, and provides for gaps in the 119915P877PC 23.12.2024 Numab Therapeutics AG sequences. Thus, in a given antibody variable region usually not all positions 1 to 149 will be occupied by an amino acid residue.
  • binding specificity refers to the ability of an individual antibody to react with one antigenic determinant and not with a different antigenic determinant.
  • the term “specifically binds to” or is “specific for” refers to measurable and reproducible interactions such as binding between a target and an antibody, which is determinative of the presence of the target in the presence of a heterogeneous population of molecules including biological molecules.
  • an antibody that specifically binds to a target is an antibody that binds this target with greater affinity, avidity, more readily, and/or with greater duration than it binds to other targets.
  • specific binding is referring to the ability of the antibody to discriminate between the target of interest and an unrelated molecule, as determined, for example, in accordance with specificity assay methods known in the art.
  • an isolated antibody may be substantially free of other cellular material and/or chemicals.
  • the antibodies of the invention are monoclonal antibodies.
  • the term “monoclonal antibody” as used herein refers to antibodies that have substantially identical amino acid sequences or are derived from the same genetic source.
  • a monoclonal antibody displays a binding specificity and affinity for a particular epitope, or binding specificities and affinities for specific epitopes. 119915P877PC 23.12.2024 Numab Therapeutics AG
  • Antibodies of the invention include, but are not limited to, chimeric, human and humanized antibodies.
  • chimeric antibody refers to an antibody molecule (or antigen-binding fragment thereof) in which (a) the constant region, or a portion thereof, is altered, replaced or exchanged so that the antigen- binding site (variable region) is linked to a constant region of a different or altered class, effector function and/or species; or (b) the variable region, or a portion thereof, is altered, replaced or exchanged with a variable region having a different or altered antigen specificity.
  • a mouse antibody can be modified by replacing its constant region with the constant region from a human immunoglobulin.
  • human antibody (or antigen-binding fragment thereof), as used herein, is intended to include antibodies (and antigen-binding fragments thereof) having variable regions in which both the framework and CDR regions are derived from sequences of human origin. Furthermore, if the antibody contains a constant region, the constant region also is derived from such human sequences, e. g., human germline sequences, or mutated versions of human germline sequences.
  • the human antibodies and antigen-binding fragments thereof of the invention may include amino acid residues not encoded by human sequences (e.
  • human antibodies can be produced using various techniques known in the art, including phage-display libraries (Hoogenboom and Winter, J. Mol. Biol, 227:381 (1992); Marks et al, J. Mol. Biol, 222:581 (1991)). Also available for the preparation of human monoclonal antibodies are methods described in Cole et al, Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p.77 (1985); Boemer et al, J.
  • Human antibodies can be prepared by administering the antigen to a transgenic animal that has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled, e. g., immunized xenomice (see, e. g., U.S. Pat. Nos.6,075,181 and 6,150,584 regarding XENOMOUSETM technology). See also, for example, Li et al, Proc. Natl. Acad. Sci.
  • humanized antibody refers to an antibody (or antigen-binding fragment thereof) that retains the reactivity of a non-human antibody while being less immunogenic in humans. This can be achieved, 119915P877PC 23.12.2024 Numab Therapeutics AG for instance, by retaining the non-human CDR regions and replacing the remaining parts of the antibody with their human counterparts (i. e., the constant region as well as the framework portions of the variable region).
  • the antibody or antigen-binding fragment thereof comprises in CDR1 of VH and/or in CDR2 of VL one or two amino acid residues that differ from the amino acid residues of the parental antibody.
  • the humanized antibodies of the invention may include amino acid residues not encoded by human sequences (e.
  • human engineering technology examples include but are not limited to the Xoma technology disclosed in U.S. Pat. No.5,766,886.
  • the term “recombinant humanized antibody” as used herein includes all human antibodies that are prepared, expressed, created or isolated by recombinant means, such as antibodies isolated from a host cell transformed to express the humanized antibody, e. g., from a transfectoma, and antibodies prepared, expressed, created or isolated by any other means that involve splicing of all or a portion of a human immunoglobulin gene, sequences to other DNA sequences.
  • the multispecific antibodies of the invention are humanized.
  • the multispecific antibodies of the invention are humanized and comprises rabbit derived CDRs.
  • the term “multispecific antibody” as used herein refers to an antibody that binds to two or more different epitopes on at least two or more different targets (e. g., CD47, PDL1, hSA and LILRB2).
  • the multispecific antibodies of the invention are bispecific or trispecific.
  • the term “bispecific antibody” as used herein refers to an antibody that binds to at least two different epitopes on two different targets (e. g., CD47 and PDL1).
  • the term “trispecific antibody” as used herein refers to an antibody that binds to at least three different epitopes on three different targets (e.
  • epitope means a protein determinant capable of specific binding to an antibody. Epitopes usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three-dimensional structural 119915P877PC 23.12.2024 Numab Therapeutics AG characteristics, as well as specific charge characteristics. “Conformational” and “linear” epitopes are distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents. [0084] The term “conformational epitope” as used herein refers to amino acid residues of an antigen that come together on the surface when the polypeptide chain folds to form the native protein.
  • linear epitope refers to an epitope, wherein all points of interaction between the protein and the interacting molecule (such as an antibody) occurring linearly along the primary amino acid sequence of the protein (continuous).
  • the term “recognize” as used herein refers to an antibody antigen-binding fragment thereof that finds and interacts (e. g., binds) with its conformational epitope.
  • the term “avidity” as used herein refers to an informative measure of the overall stability or strength of the antibody-antigen complex. It is controlled by three major factors: antibody epitope affinity; the valency of both the antigen and antibody; and the structural arrangement of the interacting parts.
  • the multispecific antibodies of the invention comprise one CD47 biding domain.
  • CD47 refers in particular to human CD47 with UniProt ID number Q08722.
  • the CD47-BD of the present invention targets CD47, in particular human CD47.
  • the multispecific antibodies of the invention comprises one CD47-BD that target human and cynomolgus (Macaca fascicularis) CD47.
  • the CD47-BD when in scFv format, binds to human CD47 with a monovalent dissociation constant (KD) of 1 to 100 nM, particularly with a KD of 2 to 50 nM, particularly of 5 to 25 nM, as measured by surface plasmon resonance (SPR).
  • KD monovalent dissociation constant
  • SPR surface plasmon resonance
  • Binding affinity generally refers to the strength of the total sum of non-covalent interactions between a single binding site of a molecule (e. g., of an antibody) and its binding partner (e. g., an antigen or, more specifically, an epitope on an antigen).
  • binding affinity refers to intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e. g., an antibody fragment and an antigen).
  • the affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (KD).
  • Affinity can be measured by 119915P877PC 23.12.2024 Numab Therapeutics AG common methods known in the art, including those described herein.
  • Low-affinity antibodies generally bind antigens slowly and tend to dissociate readily, whereas high-affinity antibodies generally bind antigens faster and tend to remain bound longer.
  • a variety of methods of measuring binding affinity are known in the art, any of which can be used for purposes of the present invention. Specific illustrative and exemplary embodiments for measuring binding affinity, i. e. binding strength are described in the following.
  • Kassoc Ka or “Kon”, as used herein, are intended to refer to the association rate of a particular antibody-antigen interaction
  • Kdis Kd
  • Koff is intended to refer to the dissociation rate of a particular antibody- antigen interaction.
  • KD is intended to refer to the dissociation constant, which is obtained from the ratio of Kd to Ka (i. e. Kd/Ka) and is expressed as a molar concentration (M).
  • M molar concentration
  • the “KD” or “KD value” or “KD” or “KD value” according to this invention is in one embodiment measured by using surface plasmon resonance assays.
  • Affinity to recombinant human CD47 and recombinant cynomolgus CD47 was determined by surface plasmon resonance (SPR) measurements, as described in Example 2.6 (scFvs) and Examples 4.2 and 5.3 (multispecific molecules).
  • Affinity to recombinant human PDL1 and recombinant cynomolgus PDL1 was determined by surface plasmon resonance (SPR) measurements, as described in Example 1.4 (scFvs) and Examples 4.2 and 5.3 (multispecific molecules).
  • the multispecific antibody of the invention acts as a blocker of the CD47 interaction with SIRP ⁇ .
  • the CD47-BD of the multispecific antibody of the invention is an inhibitor of CD47 signaling.
  • blocker or “inhibitor” or “antagonist”, as used herein, refers to an antibody or binding domain thereof that inhibits or reduces a biological activity of the antigen it binds to.
  • the CD47-BD of the multispecific antibody of the invention bind to CD47, thereby blocking the binding of CD47 to SIRP ⁇ , which leads to reduced CD47 function.
  • the CD47-BD of the multispecific antibody of the present invention is further characterized by one or more of the following parameters: a. when in scFv format, has a melting temperature (Tm), determined by differential scanning fluorimetry, in the range of 50°C to 70°C, particularly in the range of 55°C to 65°C, in particular wherein said antibody is formulated in 50 mM phosphate-citrate buffer (NaCiP) at pH 6.4, 150 mM NaCl; 119915P877PC 23.12.2024 Numab Therapeutics AG b.
  • Tm melting temperature
  • the antibody of the invention when in scFv format, has a loss in monomer content, after storage for four weeks at ⁇ 80°C, of less than 3 %, less than 2 %, less than 1 %, particularly less than 0.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer (NaCiP) with 150 mM NaCl at pH 6.4; c.
  • NaCiP phosphate citrate buffer
  • the antibody of the invention when in scFv format, has a loss in monomer content, after storage for four weeks at 4°C, of less than 3 %, less than 2 %, less than 1 %, particularly less than 0.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer (NaCiP) with 150 mM NaCl at pH 6.4; and/or d.
  • NaCiP phosphate citrate buffer
  • the midpoint of transition for the thermal unfolding of the scFv constructs is determined by using the fluorescence dye SYPRO Orange (see Wong & Raleigh, Protein Science 25 (2016) 1834-1840).
  • samples in phosphate-citrate buffer at pH 6.4 are prepared at a final protein concentration of 50 ⁇ g/ml and containing a final concentration of 5x SYPRO Orange in a total volume of 100 ⁇ l.
  • Twenty-five microliters of prepared samples are added in triplicate to white-walled AB gene PCR plates.
  • the assay is performed in a qPCR machine used as a thermal cycler, and the fluorescence emission is detected using the software’s custom dye calibration routine.
  • the PCR plate containing the test samples is subjected to a temperature ramp from 25°C to 96°C in increments of 1°C with 30 s pauses after each temperature increment.
  • the total assay time is about 2 h.
  • the Tm is calculated by the software GraphPad Prism using a mathematical second derivative method to calculate the inflection point of the curve.
  • the reported Tm is typically an average of three measurements.
  • the loss in monomer content is as determined by area under the curve calculation of SE-HPLC chromatograms.
  • SE-HPLC is a separation technique based on a solid stationary phase and a liquid mobile phase as outlined by the US Pharmacopeia (USP), chapter 621.
  • This method separates molecules based on their size and shape utilizing a hydrophobic stationary phase and aqueous mobile phase.
  • the separation of molecules is occurring between the void volume (V 0 ) and the total permeation volume (V T ) of a specific 119915P877PC 23.12.2024 Numab Therapeutics AG column.
  • Measurements by SE-HPLC are performed on a Chromaster HPLC system (Hitachi High-Technologies Corporation) equipped with automated sample injection and a UV detector set to the detection wavelength of 280 nm.
  • the equipment is controlled by the software EZChrom Elite (Agilent Technologies, Version 3.3.2 SP2) which also supports analysis of resulting chromatograms.
  • the PDL1-BD of the multispecific antibodies of the invention targets, in particular human PDL1.
  • the multispecific antibodies of the invention comprise one PDL1-BD that targets human and cynomolgus (Macaca fascicularis) PDL1.
  • the PDL1-BD used in the present invention is characterized by one or more of the following parameters: a. when in scFv format, binds to human PDL1 with a monovalent dissociation constant (KD) of 1 to 100 pM, particularly with a KD of 1 to 50 pM, particularly of 1 to 40 pM, as measured by surface plasmon resonance (SPR); b.
  • KD monovalent dissociation constant
  • the PDL1-BD of the multispecific antibodies of the present invention is further characterized by one or more of the following parameters: a.
  • Tm melting temperature
  • the antibody of the invention when in scFv format, has a loss in monomer content, after five consecutive freeze-thaw cycles, of less than 5 %, preferably less than 3 %, more preferably less than 1 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein the antibody of the invention is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4; and/or c. when in scFv format, has a loss in monomer content, after storage for four weeks at 4°C, of less than 15 %, e. g.
  • the PDL1-BDs of the multispecific antibodies of the invention are binding domains provided in the present disclosure.
  • the PDL1-BDs of the multispecific antibodies of the invention include, but are not limited to, the humanized PDL1-BDs whose sequences are listed in Table 1.
  • multivalent antibody refers to a single binding molecule with more than one valency, where “valency” is described as the number of antigen-binding moieties that binds to epitopes on target molecules.
  • the single binding molecule can bind to more than one binding site on a target molecule and/or to more than one target molecule due to the presence of more than one copy of the corresponding antigen-binding moieties.
  • multivalent antibodies include, but are not limited to bivalent antibodies, trivalent antibodies, tetravalent antibodies, pentavalent antibodies, hexavalent antibodies, and the like.
  • the term “monovalent antibody”, as used herein, refers to an antibody that binds to a single target molecule, and more specifically to a single epitope on a target molecule.
  • binding domain or “monovalent binding domain”, as used herein, refers to a binding domain that binds to a single epitope on a target molecule.
  • the multispecific antibodies of the invention comprise one CD47-BD and one PDL1-BD, i. e. the multispecific antibodies of the invention are monovalent for both CD47 and PDL1 specificity.
  • the KD of the PDL1-BD for binding to human PDL1 is at least 500 fold lower, in particular at least 1000 fold lower, in particular at least 1500 fold lower, in particular at least 2000 fold lower, than the KD of the CD47-BD for binding to human CD47.
  • the multispecific antibodies of the invention do not trigger any visible red blood cell agglutination at a concentration of 5 nM, particularly of 20 nM, particularly of 50 nM, particularly of 100 nM, particularly of 200 nM, particularly of 500 nM, as determined in a standard hemagglutination assay as described in Example 4.2
  • hemagglutination refers to agglutination of RBCs that is caused by antibodies that bind to surface antigens on the RBCs. Hemagglutination is thus a sensitive method to detect RBC- binding antibodies. The assay procedure to detect such hemagglutination is described in Example 4.2. [0111] Likewise, the multispecific antibodies of the invention do not cause the lysis of red blood cells at a concentration of 5 nM, particularly of 20 nM, particularly of 50 nM, as determined in a standard phagocytosis assay, as described in Example 5.4.
  • Antibody-dependent cell-mediated cytotoxicity (ADCC) of red blood cells is typically determine analogous to the ADCC of other cells where antibodies can bind to. More specifically, the red blood cell lysis was determined analogous to the lysis of the CD47 expressing or CD47/PDL1-coexpressing cell, as described in Example 5.4.
  • the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i.
  • the HCDR1 sequence of SEQ ID NO: 1 the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequences of SEQ ID NO: 7; or ii.
  • the HCDR1 sequence of SEQ ID NO: 20 the HCDR2 sequences of SEQ ID NO: 21, 119915P877PC 23.12.2024 Numab Therapeutics AG the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequences of SEQ ID NO: 27; or ii. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 21, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequences of SEQ ID NO: 27; or iii.
  • the HCDR1 sequence of SEQ ID NO: 20 the HCDR2 sequences of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequences of SEQ ID NO: 27; or iv. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequences of SEQ ID NO: 27; or v.
  • the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequences of SEQ ID NO: 7; or ii.
  • a binding domain used in the invention comprises a VH domain belonging to the VH3 family.
  • the term “belonging to VHx family” or “selected from VHx family” means that the respective heavy chain framework sequence or sequences referred to, e. g. FR1, FR2, FR3, FR4, FR1 to FR3, FR1 to FR4, show the highest degree of homology to said VHx family.
  • Vx family or “selected from Vx family” means that the respective light chain framework sequence or sequences referred to, e. g. FR1, FR2, FR3, FR4, FR1 to FR3, FR1 to FR4, show the highest degree of homology to said Vx family.
  • VH and VL families are given in Knappik et al., J. Mol. Biol.296 (2000) 57-86, or in WO 2019/057787.
  • a specific example of a VH domain belonging to VH3 family is represented by SEQ ID NOs: 151 and 152
  • a specific example of a VH domain belonging to VH4 family is represented by SEQ ID NO: 155.
  • heavy chain framework regions FR1 to FR4 are selected from SEQ ID NOs: 151 and 152 (Table 2, regions marked in non-bold).
  • the binding domains of the multispecific antibodies of the invention comprise a VL domain listed in Tables 1 and 2.
  • the binding domains of the multispecific antibodies of the invention comprise a VL amino acid sequence listed in Tables 1 and 2, wherein no more than about 20 amino acids in the framework sequences (i. e., the 119915P877PC 23.12.2024 Numab Therapeutics AG sequences which are not CDR sequences) have been mutated (wherein a mutation is, as various non-limiting examples, an addition, substitution or deletion).
  • the binding domains of the multispecific antibodies of the invention comprise a VL amino acid sequence listed in Tables 1 and 2, wherein no more than about 15 amino acids, particularly no more than 10 amino acids, particularly no more than 5 amino acids in the framework sequences (i. e., the sequences which are not CDR sequences) have been mutated (wherein a mutation is, as various non-limiting examples, an addition, substitution or deletion).
  • binding domains used in the invention include amino acids that have been mutated, yet have at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identity in the VL regions with a VL region depicted in the sequences described in Tables 1 and 2, including VL domains comprising at least positions 5 to 140 (AHo numbering), particularly at least positions 3 to 145 of one of the sequences shown in Tables 1 and 2.
  • the term “binding domain used in the present invention” relates to a binding domain as such, i. e. independent of a multispecific context, and, in particular, to a binding domain comprised in a multispecific construct, e. g.
  • anti-CD47xPDL1 multispecific antibodies of the invention are PRO4673, PRO4673_without_dummy_LILRB2-BD, PRO3790, PRO4688, PRO4688_without_dummy_LILRB2-BD, PRO5053, PRO5053_without_dummy_LILRB2-BD, PRO5056 and PRO5056_without_dummy_LILRB2- BD, whose sequences are listed in Tables 3 and 4.
  • the multispecific antibody additionally comprises one further binding domain, which has specificity for an antigen different from CD47 and PDL1, wherein said one further binding domain is attached to one of the C-termini of the heavy chains of the immunoglobulin Fc region.
  • the multispecific antibody additionally comprises two further binding domains, which have specificity for an antigen different from CD47 and PDL1, wherein said two further binding domains are attached to each of the C-termini of the heavy chains of the immunoglobulin Fc region.
  • the one or two further binding domains specifically bind to a tumor associated antigen (TAA) different from CD47 and PDL1. Suitable TAAs are those known in the art.
  • TAAs examples include ADRB3, AFP, ALK, BCMA, beta human chorionic gonadotropin, CA-125 (MUC16), CAIX, CD123, CD133, CD135, CD135 (FLT3), CD138, CD171, CD19, CD20, CD22, CD24, CD276, CD33, CD33, CD38, CD44v6, CD79b, CD97, CDH3 (cadherin 3), CEA, CEACAM6, CLDN6, CLEC12A (CLL1), CSPG4, CYP1B1, EGFR, EGFRvlll, EPCAM, EPHA2, Ephrin B2, ERBBs (e. g.
  • ERBB2 ERBB2
  • FAP FAP
  • FGFR1 folate receptor alpha
  • folate receptor beta Fos-related antigen
  • GA733, GD2, GD3, GFRalpha4 119915P877PC 23.12.2024 Numab Therapeutics AG globoH, GPC3, GPR20, GPRC5D, HAVCR1, Her2/neu (HER2), HLA-A2, HMWMAA, HPV E6 or E7, human telomerase reverse transcriptase, IL-11Ra, IL-13Ra2, intestinal carboxyl esterase, KIT, Legumain, LewisY, LMP2, Ly6k, MAD-CT-1, MAD-CT-2, ML-IAP, MN-CA IX, MSLN, MUC1, mut hsp 70-2, NA-17, NCAM, neutrophil elastase, NY-BR-1, NY-ESO-1, o- acetyl-GD2, OR51E2, PANX3, PDGFR
  • the one or two further binding domains specifically bind to an immunomodulatory antigen different from PDL1.
  • Suitable immunomodulatory antigen different from PDL1 are those known in the art. Suitable immunomodulatory antigen different from PDL1 are for example selected from B7-1 (CD80), B7-2 (CD86), B7-DC (CD273), B7-H1 (CD274), B7-H2 (CD275), B7-H3 (CD276), B7-H4 (VTCN1), B7-H5 (VISTA), BTLA (CD272), 4-1BB (CD137), CD137L, CD24, CD27, CD28, CD38, CD40, CD40L (CD154), CD54, CD59, CD70, CTLA-1, CTLA-4 (CD152), CXCL9, GITR (CD357), GITR, HVEM (CD270), ICAM-1 (CD54), ICOS (CD278), LAG-3 (CD223), OX40 (CD134),
  • the one or two further binding domains specifically bind to an NK cell receptor.
  • Suitable NK cell receptors are those known in the art. Suitable NK cell receptors are for example selected from KIR2DL1, KIR2DL2/3, KIR2DL4, KIR2DL5A, KIR2DL5B, KIR3DL1, KIR3DL2, KIR3DL3, KIR2DS4, KIR3DS, NKG2A, NKG2C, NKG2D, NKG2E, NKp30, NKp44, NKp46, CD2, CD16, CD69, DNAX accessory molecule-1 (DNAM-1), 2B4, NK1.1; a killer immunoglobulin (Ig)-like activating receptors (KAR); ILTs/LIRs (CD85) such as LILRA1, LILRA2, LILRA3, LILRA4, LILRA5, LILRA6, LILRB1, LILRB2, LILRB3, LILRB4, LILRB5, ILT8, ILT10,
  • the one or two further binding domains are independently of each other selected from NKp46, LILRB1, LILRB2, LILRB3, LILRB4 and LILRB5, particularly LILRB1, LILRB2, and LILRB4, particularly LILRB1 and LILRB2, particularly LILRB2.
  • the multispecific antibody comprises one further binding domain, which specifically binds to LILRB2 (LILRB2-BDs), wherein said LILRB2-BD is attached to one of the C-termini of the heavy chains of the immunoglobulin Fc region.
  • the multispecific antibody comprises two further binding domains, which specifically bind to LILRB2 (LILRB2-BDs), wherein said two LILRB2- BDs are attached to each of the C-termini of the heavy chains of the immunoglobulin Fc 119915P877PC 23.12.2024 Numab Therapeutics AG region.
  • LILRB2-BD either bind the same or different epitopes on the extracellular domain of LILRB2.
  • the two LILRB2-BDs are identical.
  • leukocyte immunoglobulin-like receptor subfamily B member 2 also known as immunoglobulin like transcript 4 (ILT4)
  • LILRB2 belongs to the subfamily B class of LIR receptors and is expressed on immune cells, such as myeloid cells, including monocytes, macrophages, dendritic cells and granulocytes. It binds classical and non- classical major histocompatibility complex Class I (MHC-I) proteins on antigen-presenting cells, such as human leukocyte antigen A (HLA-A) and human leukocyte antigen G (HLA-G), and transduces a negative signal that has immunosuppressive effects.
  • MHC-I major histocompatibility complex Class I
  • angiopoietin-like proteins such as ANGPTL2 that promotes adaptive inflammation in tissue.
  • HLA-G a nonclassical MHC-I molecule, which has been shown to mediate immunotolerance in cancer through the interaction with LILRB2, and thus allows such tumor cells to escape innate and adaptive immune response. Blocking of LILRB2 would hinder this escape mechanism. Besides, the blocking of LILRB2 can shift suppressed macrophages (M2) to activated state (M1-like).
  • M2 suppressed macrophages
  • M1-like activated state
  • LILRB2-BDs in the form of antibody fragments such as scFvs
  • scFvs anti-LILRB2 antibody building blocks
  • said building blocks should have superior biophysical properties, such as in particular a high stability, in order to facilitate their efficient incorporation into multispecific antibodies.
  • the LIRB2-BDs of the multispecific antibodies of the invention target human and cynomolgus (Macaca fascicularis) LILRB2.
  • the LILRB2-BDs used in the present invention are characterized by one or more of the following parameters: a. when in scFv format, bind to human LILRB2 with a monovalent dissociation constant (KD) of 10 pM to 5 nM, particularly with a KD of 20 pM to 3 nM, particularly of 50 pM to 1 nM, as measured by surface plasmon resonance (SPR); b.
  • KD monovalent dissociation constant
  • LILRB2 block the interaction of LILRB2 to its ligand HLA-G with an IC50 of 100 pM to 10 nM, particularly of 100 pM to 5 nM, particularly of 100 pM to 2 nM, as measured by flow cytometry; c. block the interaction of LILRB2 to its ligand HLA-A3 with an IC50 of 100 pM to 10 nM, particularly of 100 pM to 5 nM, particularly of 100 pM to 2 nM, as measured by flow cytometry; d. are not cross-reactive with members of leukocyte immunoglobulin-like receptor subfamily A (LILRA); and/or e.
  • LILRA leukocyte immunoglobulin-like receptor subfamily A
  • leukocyte immunoglobulin-like receptor subfamily A refers to leukocyte immunoglobulin-like receptor subfamily A, which, like the LILRBs, are a family of immunoreceptors that belong to leukocyte Ig-like receptors (LIRs) and that are expressed predominantly on monocytes and B cells and at lower levels on dendritic cells and natural killer (NK) cells.
  • LIRs leukocyte Ig-like receptors
  • NK dendritic cells and natural killer cells.
  • LILRA1, LILRA2, LILRA3, LILRA4, LILRA5 and LILRA6 are examples of leukocyte immunoglobulin-like receptor subfamily A, which, like the LILRBs, are a family of immunoreceptors that belong to leukocyte Ig-like receptors (LIRs) and that are expressed predominantly on monocytes and B cells and at lower levels on dendritic cells and natural killer (NK) cells. Examples are LILRA1, LILRA2, LILRA3, LILRA4, LIL
  • LILRB members other than LILRB2 refer to members of leukocyte immunoglobulin-like receptor subfamily B different from LILRB2.
  • the LILRB2-BDs of the multispecific antibodies of the present invention are further characterized by one or more of the following parameters: a. when in scFv format, have a melting temperature (Tm), determined by differential scanning fluorimetry, in the range of 55°C to 85°C, particularly in the range of 60°C to 85°C, in particular wherein said antibody is formulated in 50 mM phosphate-citrate buffer at pH 6.4, 150 mM NaCl; b.
  • Tm melting temperature
  • the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4; d. when in scFv format, have a loss in monomer content, after storage for four weeks, at 40°C, of less than 15 %, e. g.
  • the LILRB2-BDs of the multispecific antibodies of the invention are binding domains provided in the present disclosure.
  • the LILRB2-BDs of the multispecific antibodies of the invention include, but are not limited to, the humanized LILRB2-BDs whose sequences are listed in Table 1.
  • the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequences of SEQ ID NO: 7; or ii.
  • the HCDR1 sequence of SEQ ID NO: 20 the HCDR2 sequences of SEQ ID NO: 21, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, 119915P877PC 23.12.2024 Numab Therapeutics AG the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequences of SEQ ID NO: 27; or ii. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 21, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequences of SEQ ID NO: 27; or iii.
  • the HCDR1 sequence of SEQ ID NO: 20 the HCDR2 sequences of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequences of SEQ ID NO: 27; or iv. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequences of SEQ ID NO: 27; or v.
  • the HCDR1 sequence of SEQ ID NO: 112 the HCDR2 sequences of SEQ ID NO: 113, the HCDR3 sequence of SEQ ID NO: 114, the LCDR1 sequence of SEQ ID NO: 115, the LCDR2 sequence of SEQ ID NO: 116, and the LCDR3 sequences of SEQ ID NO: 118; or iv. the HCDR1 sequence of SEQ ID NO: 112, the HCDR2 sequences of SEQ ID NO: 113, the HCDR3 sequence of SEQ ID NO: 114, the LCDR1 sequence of SEQ ID NO: 115, the LCDR2 sequence of SEQ ID NO: 117, and the LCDR3 sequences of SEQ ID NO: 118 or v.
  • the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequences of SEQ ID NO: 7; or ii.
  • the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequences of SEQ ID NO: 7; or ii.
  • the HCDR1 sequence of SEQ ID NO: 93 the HCDR2 sequences of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 97, and the LCDR3 sequences of SEQ ID NO: 99; or ii.
  • the binding domains of the multispecific antibodies of the invention are selected from the group consisting of: a Fab, an scFab, an Fv, a dsFv, an scFv and a dsscFv, particularly from a Fab, Fv and scFv.
  • the binding domains of the multispecific antibodies of the invention are operably linked.
  • the binding domains of the multispecific antibodies of the invention are capable of binding to their respective antigens or receptors simultaneously.
  • the term “simultaneously”, as used herein refers for example to the simultaneous binding of the CD47-BD and the PDL1-BD or the simultaneous binding of the PDL1-BD and the LILRB2- BD.
  • the term “operably linked”, as used herein, indicates that two molecules (e. g., polypeptides, domains, binding domains) are attached in a way that each molecule retains functional activity. Two molecules can be “operably linked” whether they are attached directly or indirectly (e. g., via a linker, via a moiety, via a linker to a moiety).
  • linker refers to a peptide or other moiety that is optionally located between binding domains or antibody fragments used in the invention.
  • a number of strategies may be used to covalently link molecules together. These include, but are not limited to, polypeptide linkages between N- and C-termini of proteins or protein domains, linkage via disulfide bonds, and linkage via chemical cross-linking reagents.
  • the linker is a peptide bond, generated by recombinant techniques or peptide synthesis.
  • linker refers to a linker consisting of a chain of amino acid residues linked by peptide bonds that is connecting two domains, each being attached to one end of the linker.
  • Specific but non-limiting examples of the anti-CD47xPDL1xLILRB2 multispecific antibodies of the invention are PRO4672, PRO4687, PRO5052 and PRO5055, whose sequences are listed in table 4.
  • the multispecific antibodies of the invention can be produced using any convenient antibody-manufacturing method known in the art (see, e. g., Fischer, N.
  • bispecific antibodies comprising a functional antibody Fc part
  • methods for production of bispecific antibodies are also known in the art (see, e. g., Zhu et al., Cancer Lett.86 (1994) 127-134); and Suresh et al., Methods Enzymol.121 (1986) 210-228).
  • These methods typically involve the generation of monoclonal antibodies, for example by means of fusing myeloma cells with the spleen cells from a mouse that has been immunized with the desired antigen using the hybridoma technology (see, e. g., Yokoyama et al., Curr. Protoc. Immunol.
  • the multispecific antibodies of the invention can be prepared by conjugating the constituent binding specificities, using methods known in the art. For example, each binding specificity of the bispecific molecule can be generated separately and then conjugated to one another.
  • cross-linking agents include protein A, carbodiimide, N-succinimidyl-5-acetyl-thioacetate (SATA), 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-succinimidyl-3-(2- 119915P877PC 23.12.2024 Numab Therapeutics AG pyridyldithio)propionate (SPDP), and sulfosuccinimidyl 4-(N-maleimidomethyl)-cyclohexane- l-carboxylate (sulfo-SMCC) (see e.
  • SATA N-succinimidyl-5-acetyl-thioacetate
  • DTNB 5,5'-dithiobis (2-nitrobenzoic acid)
  • oPDM o-phenylenedimaleimide
  • SPDP pyridyldithio
  • the binding specificities are antibodies, they can be conjugated by sulfhydryl bonding of the C-terminus hinge regions of the two heavy chains.
  • the hinge region is modified to contain an odd number of sulfhydryl residues, for example one, prior to conjugation.
  • two or more binding specificities can be encoded in the same vector and expressed and assembled in the same host cell. This method is particularly useful where the bispecific molecule is a mAb x Fab, a mAb x scFv, a mAb x dsFv or a mAb x Fv fusion protein. Methods for preparing multispecific antibodies and molecules are described for example in U.S. Pat.
  • nucleic acids containing known nucleotide analogs or modified backbone residues or linkages which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides.
  • analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphorates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs).
  • PNAs peptide-nucleic acids
  • degenerate codon 119915P877PC 23.12.2024 Numab Therapeutics AG substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues (Batzer et al., Nucleic Acid Res.19:5081, 1991; Ohtsuka et al., J. Biol. Chem.260:2605- 2608, 1985; and Rossolini et al., Mol. Cell. Probes 8:91-98, 1994).
  • vectors and host cells for producing the multispecific antibody of the invention or fragments thereof or binding domains thereof.
  • vector is intended to refer to a polynucleotide molecule capable of transporting another polynucleotide to which it has been linked.
  • vector refers to a circular double stranded DNA loop into which additional DNA segments may be ligated.
  • viral vector Another type of vector is a viral vector, wherein additional DNA segments may be ligated into the viral genome.
  • Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e. g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e. g., non- episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. [0167] Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked.
  • vectors are referred to herein as “recombinant expression vectors” (or simply, “expression vectors”).
  • expression vectors of utility in recombinant DNA techniques are often in the form of plasmids.
  • 119915P877PC 23.12.2024 Numab Therapeutics AG “plasmid” and “vector” may be used interchangeably as the plasmid is the most commonly used form of vector.
  • the invention is intended to include such other forms of expression vectors, such as viral vectors (e. g., replication defective retroviruses, adenoviruses and adeno- associated viruses), which serve equivalent functions.
  • operably linked refers to a functional relationship between two or more polynucleotide (e. g., DNA) segments. Typically, it refers to the functional relationship of a transcriptional regulatory sequence to a transcribed sequence.
  • a promoter or enhancer sequence is operably linked to a coding sequence if it stimulates or modulates the transcription of the coding sequence in an appropriate host cell or other expression system.
  • promoter transcriptional regulatory sequences that are operably linked to a transcribed sequence are physically contiguous to the transcribed sequence, i. e., they are cis-acting.
  • Non-viral vectors and systems include plasmids, episomal vectors, typically with an expression cassette for expressing a protein or RNA, and human artificial chromosomes (see, e. g., Harrington et al., Nat Genet.15:345, 1997).
  • Cultures of transformed organisms can be expanded under non- inducing conditions without biasing the population for coding sequences whose expression products are better tolerated by the host cells.
  • other regulatory elements may also be required or desired for efficient expression of a multispecific antibody 119915P877PC 23.12.2024 Numab Therapeutics AG chain or a fragment. These elements typically include an ATG initiation codon and adjacent ribosome-binding site or other sequences.
  • the efficiency of expression may be enhanced by the inclusion of enhancers appropriate to the cell system in use (see, e. g., Scharf et al., Results Probl. Cell Differ.20: 125, 1994; and Bittner et al., Meth.
  • the SV40 enhancer or CMV enhancer may be used to increase expression in mammalian host cells.
  • Vectors to be used typically encode the multispecific antibody light and heavy chain or chains including constant regions or parts thereof. Such vectors allow expression of the variable regions as fusion proteins with the constant regions thereby leading to production of intact antibodies and antigen-binding fragments thereof. Typically, such constant regions are human.
  • the term “recombinant host cell” (or simply “host cell”) refers to a cell into which a recombinant expression vector has been introduced. It should be understood that such terms are intended to refer not only to the particular subject cell but to the progeny of such a cell.
  • mammalian host cells are used to express and produce the multispecific antibody of the invention.
  • they can be either a hybridoma cell line expressing endogenous immunoglobulin genes or a mammalian cell line harboring an exogenous expression vector.
  • suitable host cell lines capable of secreting intact immunoglobulins have been developed including the CHO cell lines, various 119915P877PC 23.12.2024 Numab Therapeutics AG COS cell lines, HeLa cells, myeloma cell lines, transformed B-cells and hybridomas.
  • Expression vectors for mammalian host cells can include expression control sequences, such as an origin of replication, a promoter, and an enhancer (see, e. g., Queen, et al., Immunol. Rev. 89:49-68, 1986), and necessary processing information sites, such as ribosome-binding sites, RNA splice sites, polyadenylation sites, and transcriptional terminator sequences.
  • expression control sequences such as an origin of replication, a promoter, and an enhancer (see, e. g., Queen, et al., Immunol. Rev. 89:49-68, 1986)
  • necessary processing information sites such as ribosome-binding sites, RNA splice sites, polyadenylation sites, and transcriptional terminator sequences.
  • These expression vectors usually contain promoters derived from mammalian genes or from mammalian viruses.
  • Suitable promoters may be constitutive, cell type-specific, stage- specific, and/or modulatable or regulatable.
  • Useful promoters include, but are not limited to, the metallothionein promoter, the constitutive adenovirus major late promoter, the dexamethasone-inducible MMTV promoter, the SV40 promoter, the MRP pol III promoter, the constitutive MPS V promoter, the tetracycline-inducible CMV promoter (such as the human immediate-early CMV promoter), the constitutive CMV promoter, and promoter- enhancer combinations known in the art. [0174] Methods for introducing expression vectors containing the polynucleotide sequences of interest vary depending on the type of cellular host.
  • cell lines which stably express the multispecific antibody of the invention can be prepared using expression vectors of the invention which contain viral origins of replication or endogenous expression elements and a selectable marker gene.
  • the present invention thus provides a method of producing the antibody of the invention or antigen- binding fragment thereof, wherein said method comprises the step of culturing a host cell comprising a nucleic acid or a vector encoding the antibody of the invention or antigen- 119915P877PC 23.12.2024 Numab Therapeutics AG binding fragment thereof, whereby said antibody of the disclosure or a fragment thereof is expressed.
  • the present invention relates to a method of producing the multispecific antibody of the invention, the method comprising the step of culturing a host cell expressing a nucleic acid or two nucleic acids or three nucleic acids encoding the multispecific antibody of the invention.
  • the present invention relates to a method of producing the multispecific antibody of the invention, the method comprising (i) providing a nucleic acid or two nucleic acids or three nucleic acids encoding the multispecific antibody of the invention or a vector or two vectors or three vectors encoding the multispecific antibody of the invention, expressing said nucleic acid or nucleic acids, or said vector or vectors, and collecting said multispecific antibody from the expression system, or (ii) providing a host cell or host cells expressing a nucleic acid or nucleic acids encoding the multispecific antibody of the invention, culturing said host cell or said host cells; and collecting said multispecific antibody from the cell culture.
  • the present invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising the multispecific antibody of the invention, and a pharmaceutically acceptable carrier.
  • “Pharmaceutically acceptable carrier” means a medium or diluent that does not interfere with the structure of the antibodies.
  • Pharmaceutically acceptable carriers enhance or stabilize the composition, or facilitate preparation of the composition.
  • Pharmaceutically acceptable carriers include solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible.
  • compositions for example, tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspension and lozenges for the oral ingestion by a subject.
  • Certain of such carriers enable pharmaceutical compositions to be formulated for injection, infusion or topical administration.
  • a pharmaceutically acceptable carrier can be a sterile aqueous solution.
  • the pharmaceutical composition of the invention can be administered by a variety of methods known in the art. The route and/or mode of administration vary depending upon the desired results. Administration can be intravenous, intramuscular, intraperitoneal, or subcutaneous, or administered proximal to the site of the target.
  • the administration is intramuscular, or subcutaneous, particularly subcutaneous.
  • the pharmaceutically acceptable carrier should be suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e. g., by injection or infusion), particularly for intramuscular or subcutaneous administration.
  • the active compound i. e., the multispecific antibody of the invention
  • the pharmaceutical compositions of the invention can be prepared in accordance with methods well known and routinely practiced in the art. See, e.
  • compositions are preferably manufactured under GMP conditions.
  • a therapeutically effective dose or efficacious dose of the multispecific antibody of the invention is employed in the pharmaceutical compositions of the invention.
  • the multispecific antibodies of the invention are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those of skill in the art. Dosage regimens are adjusted to provide the optimum desired response (e. g., a therapeutic response).
  • Dosage unit form refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
  • Actual dosage levels of the active ingredients in the pharmaceutical compositions of the invention can be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.
  • the selected dosage level depends upon a variety of pharmacokinetic factors including the activity of the particular compositions of the present invention employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors.
  • the multispecific antibody of the invention is usually administered on multiple occasions. Intervals between single dosages can be weekly, monthly or yearly. Intervals can also be irregular as indicated by measuring blood levels of the multispecific antibody of the invention in the patient.
  • the multispecific antibody of the invention can be administered as a sustained release formulation, in which case less frequent administration is required. Dosage and frequency vary depending on the half-life of the antibody in the 119915P877PC 23.12.2024 Numab Therapeutics AG patient. In general, humanized antibodies show longer half-life than that of chimeric antibodies and nonhuman antibodies.
  • the dosage and frequency of administration can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, a relatively low dosage is administered at relatively infrequent intervals over a long period of time. Some patients continue to receive treatment for the rest of their lives.
  • the pharmaceutical composition comprises: a) a first multispecific antibody that specifically binds to CD47 and PDL1; b) a second antibody that specifically binds to LILRB2; and c) a pharmaceutically acceptable carrier.
  • the first multispecific antibody that specifically binds to CD47 and PDL1 is selected from anti-CD47xPDL1 antibodies that are currently tested or have been tested in clinical trials.
  • antibodies that have been tested in clinical trials refers to antibodies that have successfully passed clinical testing and also to antibodies that failed in clinical testing.
  • the expression “antibodies that are currently tested in clinical trials” refers to antibodies that entered clinical trials but have not yet passed clinical trials or have not been withdrawn from clinical trials.
  • Specific non limiting examples of anti-CD47xPDL1 antibodies that are currently tested or have been tested in clinical trials are: IBI-322 by Innovent Biologics Inc.; PF- 07257876 by Pfizer Inc.; 6MW-3211 by Mabwell (Shanghai) Bioscience Co.
  • the first multispecific antibody that specifically binds to CD47 and PDL1 is selected from an anti-CD47xPDL1 multispecific antibody according to the invention, as defined herein.
  • the second antibody is a fragment-based antibody further comprising one hSA-binding domain.
  • the second antibody that specifically binds to LILRB2 is selected from anti-LILRB2 antibodies that are currently tested or have been tested in clinical trials.
  • anti-LILRB2 antibodies that are currently tested or have been tested in clinical trials are: JTX-8064 by Jounce Therapeutics Inc.; MK-4830 by Merck & Co.
  • the second antibody that specifically binds to LILRB2 is selected from antibodies that comprise at least one LILRB2-BD as defined herein.
  • the second antibody that specifically binds to LILRB2 is selected from antibodies that comprise two LILRB2-BD as defined herein.
  • Specific non-limiting examples of such anti-LILRB2 antibodies are PRO3603, PRO3635 and PRO4710, whose sequences are listed in Table 5.
  • the present invention relates to the multispecific antibody of the invention or the pharmaceutical composition of the invention for use as a medicament.
  • the present invention provides the multispecific antibody or the pharmaceutical composition for use in the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer.
  • the present invention provides the pharmaceutical composition for use in the manufacture of a medicament for the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer.
  • the present invention relates to the use of the multispecific antibody or the pharmaceutical composition for treating a disease, particularly a human disease, more particularly a human disease selected from cancer, in a subject in need thereof.
  • the present invention relates to a method of treating a subject comprising administering to the subject a therapeutically effective amount of the multispecific antibody of the present invention.
  • the present invention relates to a method for the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer, in a subject comprising administering to the subject a therapeutically effective amount of the multispecific antibody of the present invention.
  • a disease particularly a human disease, more particularly a human disease selected from cancer
  • subject includes human and non-human animals.
  • non-human mammals and non- mammals such as non-human primates, sheep, dog, cow, chickens, amphibians, and 119915P877PC 23.12.2024 Numab Therapeutics AG reptiles. Except when noted, the terms “patient” or “subject” are used herein interchangeably.
  • treatment refers to obtaining a desired pharmacologic and/or physiologic effect.
  • the effect may be therapeutic in terms of a partial or complete cure for a disease and/or adverse effect attributable to the disease or delaying the disease progression.
  • Treatment covers any treatment of a disease in a mammal, e. g., in a human, and includes: (a) inhibiting the disease, i. e., arresting its development; and (b) relieving the disease, i. e., causing regression of the disease.
  • the term “therapeutically effective amount” or “efficacious amount” refers to the amount of an agent that, when administered to a mammal or other subject for treating a disease, is sufficient to affect such treatment for the disease.
  • the “therapeutically effective amount” will vary depending on the agent, the disease and its severity and the age, weight, etc., of the subject to be treated.
  • the disease is a cancer selected from: a benign or especially malignant tumor, solid tumors, mesothelioma, brain cancer, kidney cancer, liver cancer, adrenal gland cancer, bladder cancer, breast cancer, stomach cancer (e.
  • gastric tumors e.g., gastric tumors
  • esophageal cancer ovarian cancer, cervical cancer, thymic cancer, choriocarcinoma, oral cancer, salivary cancer, colon cancer, rectum cancer, prostate cancer, pancreatic cancer, lung cancer (e. g., non-small cell lung cancer and small cell lung cancer), vaginal cancer, thyroid cancer, melanoma (e.
  • CD47 antibodies There are several anti-CD47 antibodies known in the art. However, the number of available CD47-BDs in the form of antibody fragments, such as scFvs, that can be used as universal building blocks for the construction of multispecific antibodies is limited.
  • the present invention relates to an antibody binding domain, having specificity for CD47 (CD47-BD), as defined herein.
  • said CD47-BD is in a format selected from an Fv, a dsFv, an scFv and a dsscFv, particularly from an Fv and an scFv.
  • anti-CD47 antibody binding domains are the scFvs with SEQ ID NOs: 16, 17, 18 and 19.
  • the present invention relates to an antibody comprising one or two, preferably one, antibody binding domains having specificity for CD47 (CD47-BD), as defined herein.
  • the present invention relates to an antibody binding domain having specificity for LILRB2 (LILRB2-BD), as defined herein.
  • said LIRB2-BD is in a format selected from an Fv, a dsFv, an scFv and a dsscFv, particularly from an Fv and an scFv.
  • the present invention relates to an antibody comprising one, two, three or four, preferably one or two, antibody binding domains having specificity for LILRB2 (LILRB2-BD), as defined herein.
  • LILRB2-BD antibody binding domains having specificity for LILRB2
  • Specific non-limiting examples of such anti-LILRB2 antibodies are PRO3603, PRO3635 and PRO4710, whose sequences are listed in table 5.
  • VSYSSFF SAVS M KNGGDGGV V L VQEH , s S SGSTTENVSSGY GS N ELSSKDKRCGSFSTA ASV S LL TGS D N M YGA L EKS GAG I S G I TG GSDK e ASE c Y n SSANPQEGAS VVTSKWWEPDS I G YQLPPD e u Q L GG GG I G SH P YKPV L QS TWSGA QACD C V S E L Y L PDV S A I A TGKPQ I A LI Y G S L L G E L GT WG I F A FK q e YV RGS EEYRCCY L WGSPHD L KGCKLYGKV S L sl .
  • G G 2 N SSA . 1 SE GS K L YV GSNHKSTFQ K .
  • SK I C S AS V I ED L MDTSWSSSGSDASPQSGAC GSVGSGAYSN QAT S 2 SY P TDAVQGGAKGGQSH P L WQS L 3 WKFQ E G L REH DK e WPQV P PVSGY V E .
  • Example 1 Generation and testing of antibodies directed against human PDL1: [0211]
  • the PDL1-BDs disclosed herein have been generated, produced and tested following the methods described in detail in the patent applications WO 2019/072868 and WO 2019/072869.
  • 1.1 Generation, identification, screening and testing [0212] Briefly, rabbits were immunized with recombinant human PDL1 extracellular domain. B cells from these immunized rabbits were then subjected to flow-cytometry-based sorting 119915P877PC 23.12.2024 Numab Therapeutics AG procedure.
  • PD1/PDL1 neutralizing high-affinity anti-PDL1 antibodies were identified and isolated through detecting their binding to PD-1 extracellular domain, immobilized on protein G beads.
  • the isolated B cells expressing the neutralizing anti-PDL1 antibodies were cultured, and the cultured supernatants collected.
  • the obtained rabbit monoclonal antibodies in each cell culture supernatant were characterized in a high- throughput ELISA for binding to recombinant human PDL1 and cynomolgus PDL1.
  • neutralization potential of the PDL1/PD-1 interaction as well as of PDL1/B7-1 interaction was determined by competition ELISA as well as by a cell-based reporter gene assay.
  • Fc-tagged human PDL2 was captured using the Human Antibody Capture kit from GE HealthCare. After each analyte injection cycle the CM5 sensor chip was regenerated, and new antigen was captured. The scFvs were injected as analyte at a concentration of 180 nM diluted in running buffer for 5 min and dissociation of the protein was allowed to proceed for 12 min. No binding to PDL2 was observed for PRO1392 (33-03-G02-sc18) and PRO1347 (37-20-B03-sc09).
  • the second group of three rabbits received 5 injections of 200 ⁇ g each through a period of 112 days.
  • the strength of the humoral immune response against the antigen was qualitatively assessed by determining the maximal dilution (titer) for the serum of each rabbit that still results in detectable binding of the polyclonal serum antibodies to the antigen.
  • Serum antibody titers against the immobilized antigen were assessed using an enzyme-linked immunosorbent assay (ELISA).
  • Binding to cynomolgus monkey CD47 was evaluated by direct ELISA as well as CELISA using transiently transfected cells expressing cynomolgus CD47. 119915P877PC 23.12.2024 Numab Therapeutics AG Binding to human CD47 by ELISA [0224]
  • hit identification i. e. the identification of B cell clones that produce antibodies binding to CD47
  • cell culture supernatants of the above 7,040 B cell clones were screened for the presence of antibodies that bind to human CD47 by ELISA. This primary hit identification procedure was performed with non-purified antibodies from the cell culture supernatants, as the scale of the high-throughput culture does not allow for purification of the individual rabbit antibodies.
  • the use of cell supernatants allows for ranking of large numbers of antibodies.
  • the ELISA method used assesses the “quantity” of rabbit IgGs bound to recombinant human CD47, however it does not provide any information regarding the affinity or the concentration of the antibodies.
  • supernatants from 1807 B cell clones produced a signal that was above background.
  • Inhibition of SIPR ⁇ binding to CD47 in a cellular assay and in a competition ELISA [0225] All 1807 human CD47-binding supernatants were further analyzed for their potential to inhibit the binding of SIPR ⁇ to CD47 expressing cells by flow-cytometry and for blockage of the interaction of human CD47 with human SIRP ⁇ in a competition ELISA.
  • a flow-cytometry based assay and a receptor ligand competition- ELISA were developed and adapted for use with B cell supernatant.
  • For the flow-cytometry based assay inhibition of binding of biotinylated SIPR ⁇ -Fc fusion protein to Raji cells expressing CD47 by B-cell supernatants was assessed by flow-cytometry. Since the assays could be performed with B cell supernatant as matrix and were sensitive and precise enough, both assays were used for screening. The inhibitory activity of each B cell supernatant was tested using single well analysis (no dose-responses were performed).
  • the extent of inhibition observed is not only dependent on the IgG properties, but also on the concentration of rabbit IgGs in the B cell supernatant.
  • 905 out of 1807 clones blocked the interaction of human CD47 and human SIRP ⁇ by more than 50 % and 811 clones inhibited above 70 % in the flow-cytometry based assay.
  • 1335 clones blocked the interaction of human CD47 and human SIPR ⁇ by more than 50 % and 1056 clones inhibited above 70% in the competition ELISA.850 hits show at least 50 % inhibition in both assays.
  • the assay was run at 37°C with a buffer containing 400 mM NaCl to allow better resolution of the off-rates and therefore improve the ranking of the antibodies.
  • the apparent dissociation (kd) and association (ka) rate constants and the apparent dissociation equilibrium constant (KD) were calculated with the analysis software (Analyzer, Bruker) using the 1:1 Langmuir binding model. 119915P877PC 23.12.2024 Numab Therapeutics AG [0234] Binding affinities to human CD47 could be measured for 656 anti-CD47 antibodies.
  • Proteins were purified from clarified harvest by affinity chromatography (Protein L and/or Protein A). If necessary, variant scFvs were polished by SE-chromatography to a final monomeric content > 95 %.
  • standard analytical methods such as ESI-MS, SE-HPLC, UV280 and SDS-PAGE were applied. 119915P877PC 23.12.2024 Numab Therapeutics AG [0240] The mass of the scFvs has for example been verified by the following ESI-MS standard method. Manufactured scFvs were 5 fold diluted with 1 % TFA.
  • the data were recorded with the MassLynx 4.2 Software (both Waters, UK). Where possible, the recorded m/z data of single peaks were deconvoluted into mass spectra by applying the maximum entropy algorithm MaxEnt1 (MaxLynx).
  • MaxEnt1 MaxLynx
  • the monomeric content of the manufactured scFvs has been determined for example by using the following standard SE-HPLC method. Five ⁇ g of the respective scFv, typically present at a concentration of from 0.1 to 10 mg/ml , were injected onto a Shodex KW402.5- 4F column using a Hitachi Chromaster HPLC system at 25°C and a flow rate of 0.35 ml/min.
  • the mobile phase was 50 mM sodium acetate, 250 mM sodium chloride in water at pH 6.0. Elution profile was monitored at 280 nm.
  • Table 12 Manufacture of scFvs Protein ID Based on clone ID Final titer [mg/L] Final purity by SE-HPLC [% monomer] PRO3703 70-26-B04-sc03 66.6 99.5 2.6 Pharmacodynamic characterization of suitable humanized anti-CD47-binding domains (scFv format) Binding potency to human CD47 [0242] Affinity of PRO3703 to human CD47 was determined by SPR analysis using an SPR 24 system (Sierra Sensor - Bruker).
  • Human CD47 was immobilized on a CMD200M SPR sensor prism chip (Xantec). Then PRO3703 was injected at concentrations of 10 nM, 2 nM and 0.4 nM on the sensor chip over spots with immobilized human CD47. An empty spot was used as reference. After each analyte cycle, the surface was regenerated with 3 M MgCl2 solution, thereby allowing the next analyte cycle to have only free ligand available.
  • Binding affinity KD was globally calculated by the Bruker SPR software based on the binding association (kon) and dissociation (koff) rate, by fitting a 1:1 Langmuir model to the curves 119915P877PC 23.12.2024 Numab Therapeutics AG obtained by the cycles with the three different concentrations of the analyte. The results are shown in Table 13.
  • Table 13 Affinity of PRO3703 to human CD47.
  • Protein ID scFv Affinity to human CD47 Based on clone ID k a (M -1 s -1 ) k d (s -1 ) K D (M) PRO3703 70-26-B04-sc03 2.44 ⁇ 0.21E+06 2.90 ⁇ 0.17E-02 1.19 ⁇ 0.034E-08 Binding to human and cynomolgus PBMCs [0243] The ability of the anti-CD47 scFvs to bind to specific cells within the subset of PBMCs was investigated. It was found that PRO3703 strongly binds to HLA-DR+ CD14+ monocytes and HLA-DR+ CD14- APCs.
  • PRO3703 exhibited no protein content loss at any tested storage temperature and time point.
  • Thermal unfolding [0246] Thermal stability of PRO3703 was analyzed by nano dynamic scanning fluorimetry (nDSF) using a NanoTemper, allowing the determination of the onset of unfolding (Tonset), midpoint of unfolding (Tm). Tm and Tonset results of duplicate/triplicate measurements are summarized in Table 17. As can be deduced from Table 17, all molecules exhibit high melting temperatures of at least 59°C, most of the molecules exhibit melting temperatures of >60°C or >70°C.
  • the quality of recombinant human LILRB2 was analyzed by the manufacturer for purity by SDS-page.
  • the first group of three rabbits (protocol 1) received 4 injections of 200 ⁇ g each throughout 33 days.
  • the second group of six rabbits (protocol 2) received 4 injections of 200 ⁇ g each throughout 70 days.
  • the third group of three rabbits (protocol 3) received 5 injections of 200 ⁇ g each through a period of 108 days.
  • the strength of the humoral immune response against the antigen was qualitatively assessed by determining the maximal dilution (titer) for the serum of each rabbit that still results in detectable binding of the polyclonal serum antibodies to the antigen.
  • n o c y a C D P ( 0 8 - a . n Tm 8 a 2 ) t f a y a C C ° . 3 e 8 2 ) o n D M ( 0 4 9 9 g a r y a C C ° 0 . 5 0 i ot 4 o a t D P ( 4 1 i n 1 ) s 4 y a C C ° . 5 l e D M ( 0 4 9 9 p 1 ) m y a C C P ° r e ( 0 4 5 .
  • the ability of the antibodies to cross-react with cyno and rhesus LILRB2 was assessed by CELISA using transiently transfected HEK293T cells expressing rhesus or cynomolgus LILRB2 with an N-terminal V5-tag. [0257] 141 (9.9 %) and 51 clones (3.6 %) showed binding to cynomolgus monkey and rhesus monkey LILRB2, respectively. Overall, there were only a few antibodies identified showing cross-reactivity to either of the two monkey species.
  • Protein ID scFv Affinity to human LILRB2 Based on clone ID ka (M -1 s -1 ) kd (s -1 ) KD (M) PRO3744 68-18-E11-sc07 7.71E+05 1.14E-04 1.48E-10
  • PRO3697 69-06-G04-sc05 9.74E+05 1.28E-04 1.31E-10 Binding of anti-LILRB2 scFvs to CHO K1 cells expressing human LILRB2 [0268] To test binding of Numab’s anti-LILRB2 scFvs PRO3696 and PRO3744 to CHO K1 cells constitutively expressing human LILRB2, a flow cytometry-based assay was developed.
  • CHO K1 cells expressing human LILRB2 were seeded in a 96-well plate (50,000 cells per well) and 3-fold serial dilutions of the molecules to test were added at concentrations starting at 100 nM to the plates and incubated for 60 min at 4°C on a shaker. Supernatants were removed and detection reagent (Numab’s anti- framework antibody conjugated to Alexa Fluor 647, in-house) was added for flow cytometry analysis. The plates were measured with the flow cytometer Novocyte with a flow rate of 120 119915P877PC 23.12.2024 Numab Therapeutics AG ⁇ l/min and the geometric mean of the fluorescence intensity of single cells was calculated.
  • Binding of anti-LILRB2 scFvs to different human and cynomolgus PBMC cell types [0271] Binding of the anti-LILRB2 scFv PRO3744 to different PBMC cell types from human as well as from cynomolgus monkeys was examined within the specified subset of T cells, B cells, and myeloid cells. In particular, the ability of anti-LILRB2 scFv PRO3744 to target HLA- DR+ CD14+ monocytes and HLA-DR+ CD14- APCs from human and cynomolgus monkey was investigated.
  • the IC 50 as well as maximum inhibition values for the anti-LILRB2 scFvs PRO3744, PRO3696 and PRO3697 were normalized to the control IgG antibody J-19 (PRO2518), which exhibits high affinity binding to human LILRB2.
  • all tested scFvs namely PRO3744, PRO3696, and PRO3697, demonstrated potent and comprehensive inhibition of the LILRB2/HLA-G interaction with potencies similar to the bivalent IgG antibody J-19 (PRO2518).
  • PRO3744 also achieved complete neutralization of the HLA-A3/LILRB2 interaction, while PRO3696 exhibited partial inhibition, and PRO3697 did not impede HLA-A3 binding.
  • LILRB2 selectivity assessment [0277] To assess whether the anti-LILRB2 antibodies selectively bind to LILRB2 and not to other members of protein family LILRA and LILRB, a cellular ELISA (CELISA) using transiently transfected CHO cells was developed. [0278] The selectivity assessment was performed as follows.
  • LILRA1 (identifier: O75019-1), LILRA2 (Q8N149-1), LILRA3 (Q8N6C8-1), LILRA4 (P59901-1), LILRA5 (A6NI73-1), LILRA6 (Q6PI73-1), LILRB1 (Q8NHL6-1), LILRB2 (Q8N423-2), LILRB3 (O75022-1), LILRB4 (Q8NHJ6-1) or LILRB5 (O75023-1).
  • PRO3696 a precursor of PRO3696 (69-01-E02-sc06), exhibited the same selectivity for LILRB2 as PRO4710, when tested at a concentration of 150 nM (see Figure 4).
  • cross-linked PRO3697 (69-06-G04-sc05) showed, besides strong binding to LILRB2, weak binding to LILRB1, but no binding to any of the tested LILRBA family members or to LILRB3, LILRB4 or LILRB5, as seen for the other clones (see Figure 5).
  • HMDM human monocyte-derived macrophages
  • PBMCs Human peripheral blood mononuclear cells
  • HMDM+ cells were isolated from PBMCs by positive selection with magnetic beads (Miltenyi Biotech, 130- 050-201). HMDMs were generated by culturing CD14+ cells for 6-8 days in complete medium (RPMI 1640, 10 % heat-inactivated bovine serum, Sigma Aldrich) in the presence of 50 ng/ml human macrophage colony stimulating factor (M-CSF) (Peprotech) at 37°C and 5 % CO2. Medium was exchanged and M-CSF replenished on day 3 of the culture. [0286] HMDM cells were exposed to 25 ng/mL IL-4 in the presence of a serial dilution of PRO3744, PRO3696, and PRO3697 at 37°C and 5 % CO2.
  • M-CSF human macrophage colony stimulating factor
  • HMDMs were harvested from culturing flasks on day 6-8 using Accutase (Biowest), seeded at 50’000 cells per well in 96-well flat bottom plates (TPP) and treated with 25 ng/mL IL-4 (Peprotech, 200-04 – 50 ⁇ g) together with a serial dilution of test compounds or isotype antibodies (dose range 100 nM – 0.0006 nM) for 24 h at 37°C and 5 % CO2. After 24 h, cells were stained with Zombie Aqua Fixable Viability Kit (BioLegend) for 30 min at 4°C.
  • Fc receptor block Human TruStain FcX, BioLegend, 422302
  • Staining antibodies All from BioLegend were added to the cells for 30 min at 4°C: CD163- APC (cat. no.333610), CD206-BV711 (cat. no.321136), CD86-AF488 (cat. no.305414), CD80-PE (cat. no.305208) and CD14-PerCp-Cy5.5 (cat. no.325622).
  • Example 4 Manufacture and characterization of anti-CD47 x PDL1 bispecific antibodies: 4.1 Manufacture of anti-CD47 x PDL1 bispecific antibodies [0292] Manufacture anti-CD47 x PDL1 bispecific antibodies was performed as follows. Expression of multispecific constructs was performed in ExpiCHO-S cells using the transient ExpiCHO expression system (Gibco). Expression cultures were cultivated in batch using shaking flasks for 8 days at 37°C. The culture supernatants were separated from cells by centrifugation followed by a 0.22 ⁇ m sterile filtration.
  • the reference PRO3609 has an (scFv)2- Fc antibody format, which is a format according to the invention, but comprises an CD47-BD that has an intermediate (higher) binding affinity. 8 2 y % ) a C C D M ° ( 0 4 4 . 0 5 f 8 . o 2 69 . 3 s y a % ) C C ° e r D P ( 0 4 . 53 - . 54 - . 72 -d 4 ) u t n 1 a y % aC C M ° a 4 ) ( 0 4 . 9 r 0 - 9 . 5 2 .
  • CRL-1848, double positive for human CD47 and PDL1) or CHO CD47 (in-house cell line, positive for human CD47) were seeded in a 96-well plate. Threefold serial dilutions of the molecules to test were added at concentrations ranging from 100 nM to 2 pM (PRO2777 on NCI-H292 and CHO CD47), from 300 nM to 5 pM (for PRO3790 and PRO3599 on NCI-H292) or from 500 nM to 9 pM (for PRO3790 and PRO3599 on CHO CD47) to the plates and incubated for 60 min at 4°C on a shaker.
  • PRO3790 and PRO3599 showed potent inhibition of CD47/SIRP ⁇ interaction to PDL1 co-expressing NCI-H292 cells, and weak inhibition of SIRP ⁇ binding to CHO CD47 cells which do not express PDL1.
  • PRO3790 showed a better potency to inhibit SIRP ⁇ binding to NCI-H292 cells than the Pfizer molecule (PRO3599), while the potency was 119915P877PC 23.12.2024 Numab Therapeutics AG comparable for the two molecules when CHO CD47 cells were used.
  • High affinity anti-CD47 molecule PRO2777 demonstrated potent inhibition of SIRP ⁇ binding to both cell lines tested.
  • Cell binding to RBCs [0304] To test the binding of PRO3790 and PRO2777 to human red blood cells (RBCs), a flow cytometry-based assay was used. [0305] Human RBCs were isolated from peripheral blood of healthy donors. Human RBCs were seeded in a 96-well plate and 3-fold serial dilutions of the molecules to test were added at concentrations ranging from 100 nM to 1.7 pM (for PRO2777) or from 500 nM to 8.5 pM (PRO3790) to the plates and incubated for 60 min at 4°C with shaking.
  • Hemagglutination To test whether PRO3790 triggers agglutination of human erythrocytes, a hemagglutination assay based on optical density measurement was performed. [0309] The hemagglutination assay was performed as follows. Human erythrocytes were isolated from peripheral blood of healthy volunteer and 1 mio. human erythrocytes were seeded in a 96-well plate.
  • PRO3790 showed no sign of hemagglutination at all tested concentrations (highest concentration was 500 nM), while the magrolimab analog (PRO2777) was able to trigger agglutination of human erythrocytes with EC50 value of 0.182 nM.
  • Table 32 Hemagglutination Hemagglutination assay Rel. maximum Rel.
  • CHO cells stably expressing PDL1 and a T cell receptor (TCR) activator were incubated with Jurkat T cells stably expressing PD-1 and as a reporter gene to monitor T cell activation, firefly luciferase under the control of the NFAT response elements (target and effector cells both from BPS Bioscience).
  • TCR activator Upon binding of the TCR activator on CHO cells to the Jurkat T cells, TCR signaling triggers NFAT-induced expression of firefly luciferase.
  • the interaction between PDL1 and PD-1 negatively regulates TCR signaling and decreases firefly luciferase expression. Consequently, blockade of the PDL1/PD-1 interaction in this system restores luciferase activity.
  • 35’000 CHO/PDL1/TCR activator cells (BPS Bioscience) in 100 ⁇ l of cell culture medium (DMEM/F12, 10 % FCS) were added to the inner wells of a white cell culture plate and incubated for 16-20 h at 37°C and 5 % CO2. On the next day, 95 ⁇ l of cell culture medium was removed from each well and 50 ⁇ l of 2-fold concentrated 3-fold serial dilutions of the multispecific antibody test compounds comprising a PDL1-BD and of reference antibody avelumab ranging from 50 nM to 8 pM (final concentration in the well) were added.
  • Concentration-response curves were fitted by using a four-parameter logistic fit (GraphPad prism software) and the half-maximal effective concentration (EC50) was calculated.
  • EC50 half-maximal effective concentration
  • relative EC50 values were used. These values were determined by calibrating the EC50 value of the molecules against the reference molecule avelumab included on each assay plate (relative EC50: EC50, avelumab / EC50, test).
  • the maximum activation of each multispecific antibody test compound was calculated and normalized to the maximum activation seen for avelumab (relative maximum activation: maximum activation, test / maximum activation, avelumab).
  • PRO3790 showed potent blockade of PD-1 / PDL1 interaction in transgenic Jurkat NFAT reporter gene assay, comparable to benchmark antibody avelumab and reference molecule PRO3599 (Table 33).
  • Table 33 PDL1 / PD-1 transgenic NFAT reporter gene assay Rel. EC50 Rel. maximum (EC 50 activation (max. PRO ID PDL1 binder CD47 binder EC50 [nM] avelumab / activation sample EC 50 / max.
  • PBMCs Human peripheral blood mononuclear cells
  • HMDMs were generated by culturing CD14+ cells for 6-8 days in complete medium (RPMI 1640, 10 % heat-inactivated bovine serum, Sigma Aldrich) in the presence of 50 ng/ml human macrophage colony stimulating factor (M-CSF) (Peprotech) at 37°C and 5 % CO2. Medium was exchanged and M-CSF replenished on day 3 of the culture.
  • complete medium RPMI 1640, 10 % heat-inactivated bovine serum, Sigma Aldrich
  • M-CSF human macrophage colony stimulating factor
  • HMDMs were harvested from culturing flasks on day 6-8 using Accutase (Biowest), seeded at 50’000 per well in 96-well flat bottom plates (TPP) and treated with 10 ng/ml IL-10 (Peprotech) overnight. The next day HMDMs were co-incubated with CFSE-labelled target cells at an effector:target ratio of 1:1 for 2.5 h at 37°C in the presence of a serial dilution of test compounds (dose range 200 nM – 0.02 pM). In the case of RBCs, the effector:target ratio was 1:50.
  • the phagocytosis curve exhibits a bi-phasic pattern, in contrast to magrolimab, which demonstrated a straightforward dose-dependent phagocytosis.
  • the initial part of the curve depends on PDL1 inhibition, whereas the second part is driven by CD47 blockade. Given that the PDL1 inhibitor is the same across all three tested molecules, the EC 50 for this phase remains comparable.
  • Phagocytosis of tumor cell by PDL1 x CD47 bi-specific antibodies is more pronounced with high affinity CD47 binder (PRO3791) when compared to those with low affinity CD47 moieties (PRO3790 and PRO3802).
  • Example 5 Anti-CD47 x PDL1x LILRB2 trispecific antibodies 5.1 Manufacture of anti-CD47 x PDL1x LILRB2 trispecific antibodies [0318]
  • the trispecific antibodies according to the present invention have been produced analogous to the methods described in Example 4.1. Briefly, expression of multispecific constructs was performed in ExpiCHO-S cells using the transient ExpiCHO expression system (Gibco). Expression cultures were cultivated in batch using shaking flasks for 8 days at 37°C. The culture supernatants were separated from cells by centrifugation followed by a 0.22 ⁇ m sterile filtration.
  • target proteins were captured from the clarified culture supernatants by protein A affinity chromatography and were polished by a size-exclusion chromatography (SEC) step, except for PRO3851, which was polished by cation-exchange chromatography (CIEX). 119915P877PC 23.12.2024 Numab Therapeutics AG [0319] Some target proteins with a high post-capture monomeric purity (i. e. PRO2518, PRO3483, PRO3603) were not SEC-polished and, instead, dialyzed against the respective final buffer in a dialysis membrane. For the quality control of the manufactured material, standard analytical methods such as SE-HPLC, SDS-PAGE, and UV280 were used. Table 34 shows a production overview of these molecules.
  • Freeze-thaw stability As in the early development stages the drug substance is usually stored frozen, formulation and fill/finish processing will require some freezing and thawing operations. The compatibility of the top performing trispecific molecules with respect to freeze-thawing (F/T) cycles (colloidal stability) was assessed. [0323] For the F/T stability assessment the same analytical methods (SE-HPLC, cGE) and parameters (% monomer content and % monomer loss) as for the storage stability study were applied to monitor the quality of the molecules over five F/T cycles. Table 35 illustrates that after five F/T cycles (F/T) the loss of monomer content was less than 1 %.
  • binding affinity without avidity was determined by using a proprietary anti- framework rabbit IgG (rIgG21-57-20-B11) immobilized to a carboxymethylated dextran surface (CM5 sensorchip; Biacore, Cytiva) to capture the multispecific antibodies according to the present invention on the surface.
  • CM5 sensorchip carboxymethylated dextran surface
  • Biacore Cytiva
  • Binding of trispecific antibodies to different human and cyno PBMCs cell types [0327] Binding of the multispecific antibodies of the invention to different PBMC cell types from human as well as from cynomolgus monkeys was examined within the specified subset of T cells, B cells, and myeloid cells. In particular, the ability of the anti-CD47 x PDL1 x LILRB2 antibodies to target HLA-DR+ CD14+ monocytes and HLA-DR+ CD14- APCs, both from human and cynomolgus monkey, was investigated. The method used to assess the binding to these cell-types was analogous to the method described in the preceding section. The results are summarized in Table 37.
  • Binding is detected by an increase in fluorescence signal indicated in [GeoMFI x 1000].
  • Re-polarization of M2 macrophages and LPS-driven cytokine secretion [0328] The ability of the anti-CD47 x PDL1 x LILRB2 antibodies to re-polarize M2 macrophages and to foster LPS-driven cytokine secretion was assessed using the methods disclosed in Example 3.6. 119915P877PC 23.12.2024 Numab Therapeutics AG Table 36: Affinity of different multispecific test constructs to human and cynomolgus CD47, human and cynomolgus PDL1, and human and cynomolgus LILRB2.
  • the reference anti-PDL1 x CD47 bi-specific antibodies from Pfizer (PRO3599), the LILRB2-BD-devoid trispecific variants PRO5053 and PRO5056 (HSA-BD as LILRB2 mock binding domain), as well as the anti-CD47 antibody magrolimab and the anti-PDL1 antibody atezolizumab (tested 119915P877PC 23.12.2024 Numab Therapeutics AG at a 50 nM dose), showed no re-polarization activity. Results of the cytokine secretion are summarized in Table 39.
  • Table 38 Re-polarization potency of M2 macrophages for the anti-LILRB2 IgG PRO4710, the trispecific antibodies PRO4672, PRO4687, PRO5052, PRO5053, PRO5055 and PRO5056, and reference compounds magrolimab, atezolizumab, and PRO3599 PRO ID CD86 EC 50 [nM] CD163 EC 50 [nM] CD206 EC 50 [nM] PRO4710 0.119 ⁇ 0.070 0.072 ⁇ 0.008 0.203 ⁇ 0.129 magrolimab n.a. n.a. n.a. atezolizumab n.a. n.a. PRO3599 n.a. n.a. n.a.
  • PRO4672 0.079 ⁇ 0.009 0.067 ⁇ 0.004 0.120 ⁇ 0.043
  • PRO5052 0.082 ⁇ 0.006 0.073 ⁇ 0.001 0.134 ⁇ 0.082
  • PRO5053 n.a. n.a. n.a.
  • PRO5055 0.085 ⁇ 0.012 0.058 ⁇ 0.003 0.128 ⁇ 0.071
  • PRO5056 n.a. n.a. n.a.
  • the trispecific antibodies according to the invention further exhibit the ability to foster LPS-driven cytokine secretion (see Figures 8 and 9, and Table 39), due to the presence of the LILRB2 arm, which is confirmed through comparison with the respective mock compounds.
  • the tested multispecific antibodies demonstrate similar effects obtained with the reference anti-LILRB2 antibody J19.H1 from Jounce (PRO2518, see Figure 9), while there is no observed activity for the LILRB2-BD-devoid trispecific variants PRO5053 and PRO5056 (HSA-BD as LILRB2 mock binding domain) and the reference antibodies PRO3599, magrolimab and atezolizumab (at the top dose).
  • Both assays demonstrate the efficacy of LILRB2 inhibition through the absence of activity in PDL1 x CD47 bi-specific compounds, and Pfizer compound. Furthermore, comparable activity is observed between multispecific constructs comprising a high-affinity CD47-BD and a low-affinity CD47-BD. Notably, the trispecific antibodies of the invention having different PDL1-BDs exhibit an activity comparable with established benchmarks from Jounce, Merck, and NGM.
  • Phagocytosis The in vitro phagocytic activity induced by bi-/multispecific antibodies of the invention was assessed using the H292, A375, Nalm6, tumor cell lines and primary red blood cells (RBCs). The assay relies on the uptake of labelled target cells by macrophages and analysis by flow cytometry.
  • PBMCs Human peripheral blood mononuclear cells
  • CD14+ cells were isolated from PBMCs by positive selection with magnetic beads (Miltenyi Biotech, 130- 050-201).
  • HMDMs were generated by culturing CD14+ cells for 6-8 days in complete medium (RPMI 1640, 10 % heat-inactivated bovine serum, Sigma Aldrich) in the presence of 50 ng/ml human macrophage colony stimulating factor (M-CSF) (Peprotech) at 37°C and 5 % CO 2 . Medium was exchanged and M-CSF replenished on day 3 of the culture.
  • M-CSF human macrophage colony stimulating factor
  • HMDMs were harvested from culturing flasks on day 6-8 using Accutase (Biowest), seeded at 50’000 cells per well in 96-well flat bottom plates (TPP) and treated with 10 ng/ml IL-10 (Peprotech) overnight.
  • HMDMs were co-incubated with CFSE-labelled target cells at an effector:target ratio of 1:1 for 2.5 h at 37°C in the presence of a serial dilution of test compounds (dose range 200 nM – 0.02 pM). In the case of RBCs, the effector:target ratio was 1:50.
  • cells were stained with a viability marker (Zombie NIR, Biolegend) and an anti-CD14-APC staining antibody (Biolegend). Samples were fixed and acquired by flow cytometry (Attune NXT, Thermo 119915P877PC 23.12.2024 Numab Therapeutics AG Fisher Scientific).
  • Phagocytic activity was determined as double positive events (CD14+ HMDMs that have taken up CFSE positive target cells) and phagocytosis is presented as % CFSE+ macrophages.
  • Tables 40, 41, 42 and 43 The results of the phagocytosis potency analysis are summarized in Tables 40, 41, 42 and 43 as well as in Figures 10, 11, 12 and 13.
  • the multispecific antibodies induced a bi- phasic response curve, with an initial part driven by the anti-PDL1 arm, confirmed through comparison with avelumab (anti-PDL1 IgG1). This initial increase in phagocytosis was absent in PDL1-negative cell lines, confirming PDL1 dependency.
  • the second part of the dose-response curve is driven by anti-CD47 engagement, which was confirmed by PRO5054 and PRO5057, two compounds in which anti-CD47 was replaced by an unspecific human serum albumin (hSA) binder (hSA-BD), herein also referred to as mock binding domain.
  • hSA human serum albumin
  • hSA-BD unspecific human serum albumin binder
  • the herein described antibodies showed similar potency but higher efficacy as the benchmark compounds magrolimab, PRO3599 (anti-CD47xPDL1 bispecific antibody of Pfizer), PRO3851 (anti-CD47xPDL1 bispecific antibody of Innovent) and a combination of magrolimab & avelumab.
  • phagocytosis potency may also be increased by the presence of the LILRB2-BDs, as indicated by the somewhat reduced activity observed in the trispecific molecules, where the LILRB2-BDs have been replaced by mock binding domains (PRO5053 and PRO5056).
  • LILRB2-engaging domains in proximity to the Fc-region improve orientation of the 119915P877PC 23.12.2024 Numab Therapeutics AG compounds enhancing trans-bridging of macrophages and tumor cells (in absence and presence of PDL1 expression on tumor cells).
  • Many different multispecific antibody formats have been tested for H292 cancer cell phagocytosis and red blood cells (RBC) phagocytosis (see Table 43 and also Figures 10, 11, 12, 13, 14 and 15).
  • the bispecific antibody PRO3790 and the trispecific antibodies PRO4672, PRO4673, PRO4687, PRO4688, RO5052, PRO5053, PRO5055 and PRO5056 exhibit high cancer cell phagocytosis and, at the same time, only minimal or no measurable RBC phagocytosis.
  • the bispecific and trispecific antibodies of the invention thus have a significantly larger therapeutic window than the reference antibodies PRO3599, PRO3851 and magrolimab.
  • the phagocytosis data in Table 43 further illustrates that placing the CD47- BD at the C-terminus of the Fc-region (PRO4677, PRO4678) or replacing the CD47-BD by a mock domain (PRO5054), also lead to minimal or no measurable RBC phagocytosis, but also dramatically decrease cancer cell phagocytosis.
  • all tested multispecific antibodies of the invention containing a CD47-BD in scFv format in one of the Fab arms did not induce noteworthy RBC phagocytosis at all concentrations tested and, thus, exhibit a large potential therapeutic window, as illustrated in Figure 15.
  • Phagocytosis of PDL1 positive tumor cells was unaffected by the different format and positioning of the CD47-BD, except for PRO4677, where tumor and RBC phagocytosis were abolished.
  • the reference anti-CD47 x PDL1 antibodies PRO3599 (Pfizer) and PRO3851 (Innovent), and magrolimab exhibit the highest maximal levels of RBC phagocytosis.
  • Multispecific antibodies activate NK cells to tumor cell killing via ADCC
  • the in vitro cytotoxic activity of primary NK cells induced by the multispecific antibodies according to the invention was assessed using the H226 tumor cell line, transfected with the IncuCyte NucLight Orange Lentivirus Reagent (Sartorious, 4625).
  • the assay makes use of the IncuCyte live imaging platform (Sartorius) and quantifies the killing of H226 NucRed tumor cells as measured by the loss of the NucRed signal over time.
  • PBMCs Human peripheral blood mononuclear cells
  • NK cells were subsequently isolated by negative selection with magnetic beads (Miltenyi Biotech, 130-092-657). NK cells were pre-activated overnight in (RPMI 1640, 10 % heat-inactivated bovine serum, Sigma Aldrich) in the presence of 0.2 – 1 nM human IL-2 (R&D Systems) at 37°C and 5 % CO2.
  • RPMI 1640 10 % heat-inactivated bovine serum, Sigma Aldrich
  • H226 NucRed tumor cells were seeded at 7’000 cells per well in 96-well flat bottom plates (TPP) on day -1 and cultured in RPMI+10 %FCS.
  • NK cells were harvested, washed with medium and co-incubated with target cells at an effector:target ratio of 5:1 for 24 h at 37°C in the presence of a serial dilution of test compounds (dose range 20 nM – 0.03 pM). Images of the tumor cell - NK cell co-culture were taken every two hours and the count of H226 NucRed tumor cells was assessed over time. Data is presented as fold change to T0 for each compound and is an average of four images per well. [0343] The trispecific antibodies PRO5052 and PRO5055 exhibited a potent ADCC with EC50 values that are similar to that of avelumab and higher than that of the reference PRO3599.
  • Magrolimab (anti-CD47 IgG4) and atezolizumab (anti-PDL1 modified IgG1) failed to induce ADCC.
  • the Fc-silent variant PRO5134 failed to induce ADCC.
  • the trispecific antibody PRO5054 where the CD47-BD is replaced by a mock binding domain, shows a significantly reduced ADCC, highlighting the importance of simultaneous targeting of PDL1 and CD47.
  • the presence or absence of LILRB2-BDs in the trispecific molecules had no effect on ADCC potency.
  • 5.6 Induction of T cell proliferation [0344] To evaluate whether the trispecific antibodies of the invention are capable to activate T cell proliferation, these antibodies were tested in an immunosuppressive assay.
  • the immunosuppressive assay used aims to track recovery of T cell proliferation in the co-culture 119915P877PC 23.12.2024 Numab Therapeutics AG with suppressive macrophages in the presence of T cell stimulation and test compounds.
  • the assay was performed as follows. [0345] T cells were isolated from PBMC fraction with a magnetic Pan T Cell negative isolation kit (Miltenyi Biotec, cat no.130-096-535). Bulk T cell population was used containing CD4+ and CD8+ T cells. T cells were stained with a cell proliferation tracking dye (CTV CellTrace dye, Thermo Fisher Scientific, cat no. C34557) by incubation of T cells with the CTV dye (5 ⁇ M working solution) for 20 min at 37°C.
  • CTV CellTrace dye Thermo Fisher Scientific, cat no. C34557
  • T cells were stimulated to proliferate by a co-culture in 96 flat-bottom well plates (TPP) with coated anti-CD3 (at 2 ug/mL, BioLegend, cat no.317326) and soluble anti-CD28 mAbs (4 ⁇ g/mL, BioLegend, cat no.302934).
  • Suppressive human monocyte-derived macrophages (HMDM) were generated by treatment with IL-10 (10 ng/mL, BioLegend cat no.
  • TGF- ⁇ (20 ng/mL, Peprotech, cat no.100-21) for 24 h at 5 % CO2 and 37°C prior to co-culture with T cells.
  • HMDM were added to the culture in different ratios, e. g.1:1, 2.5:1 and 5:1 (T cell : HMDM). Total number of cells varied from 120 k to 150 k per well.
  • Co- culture of T cells and macrophages lasted for 5 days at 37°C and 5 % CO2.
  • As a positive control stimulated T cells cultured without HMDM were used (max. proliferation observed).
  • T cell proliferation was measured by quantification of CTV dilution, which increases with every cell division. Data was collected with flow cytometry method, where T cells and HMDM were identified by staining with mAb (all from BioLegend: CD14-APC cat. no.325608, CD4-FITC 317408, CD8 PercP cat no.344708). Live/dead stain (Zombie NIR Fixable Viability Kit, BioLegend, cat. No.423106 ) was employed to exclude dead cells from subsequent analysis.
  • Trispecific molecules of the invention demonstrate a dose-dependent induction of T-cell proliferation, as indicated by the significant CTV dye dilution and proliferation index (see Figure 16). This heightened activity is attributed to the inhibition of LILRB2, as evidenced by the singular effectiveness of the IgG compound as a standalone agent.
  • PBMC Peripheral blood mononuclear cells
  • PBMCs were isolated from fresh human whole blood employing density gradient centrifugation.
  • PBMCs were depleted of NK cells using anti-CD56 microbeads (Miltenyi Biotec, 130-097-042) and the MACS cell separation kit (Miltenyi Biotec).200,000 NK-cell depleted PBMCs per well were added to the 96-well plate (flat bottom, TPP), followed by the addition of serial dilutions of experimental compounds in assay buffer containing Staphylococcal enterotoxin A (SEA, Toxin Technology, AT 101) at a concentration of 10 ng/mL.
  • SEA Staphylococcal enterotoxin A
  • IL-1 ⁇ induction is primarily attributed to PDL1-binding, as demonstrated by the response to atezolizumab, with no discernible impact from CD47 and LILRB2 blockade.
  • the multispecific compound exhibited activity on par with the combination of three IgGs, considering data from five donors. However, it is important to note a significant variability among donors in the observed outcomes.
  • Biphasic binding curves were obtained on HT-1080 cells, due to sequential binding of the PDL1- and CD47-BDs to the cells. For HT-29 cells, only monophasic binding curves were obtained, as these cells do not express PDL1.
  • PRO4672, PRO4687, PRO5052 and PRO5055 demonstrated potent binding to HT-1080 and weaker binding to HT-29 cells.
  • the reference antibody PRO3599 exhibited a similar binding profile to both cancer cell lines, albeit with a somewhat weaker potency for HT-29 cancer cells.
  • the reference antibodies PRO2777 and PRO3851 showed a monophasic binding profile to both cell lines with PRO3851 exhibiting the highest Emax of all tested antibodies.
  • the other reference antibodies PRO2777 (magrolimab) and PRO3851 showed similar SIRP ⁇ neutralization potencies on NCI-H292 cells. 5.11 LILRB2 selectivity assessment [0360] To assess whether the trispecific antibodies according to the invention selectively bind to LILRB2 and not to other members of protein family LILRA and LILRB, a cellular ELISA (CELISA) using transiently transfected CHO cells was developed. The method applied for this assessment was the same as described in Example 3.6. [0361] As shown in Figure 23, trispecific antibodies PRO5052 and PRO5055 demonstrated potent and selective binding to LILRB2.
  • CELISA cellular ELISA
  • This assay aimed to assess the safety profile of the multispecific antibodies according to the present invention in human whole blood by measuring immune cell count and activation, as well as cytokine release after treatment with test compounds. Both PRO5052 and PRO5055 multispecific antibodies were tested in parallel. Mock-compounds (hSA- binding domain) for each individual arm were included to assess possible mode of action in case of unexpected activity in human whole blood (see Table 44).
  • - ⁇ CD3 mAb OKT3 clone, soluble & coated
  • T-cell activator - ⁇ CD52 mAb
  • IgG1 based T and B cell depletion IgG1 based T and B cell depletion
  • - PHA Phytohemagglutinin
  • the assay setup was as follows: - Donors tested: tested in 3 independent experiments with 2 human donors per replicate - Incubation conditions: 20h, 37°C, 5% CO2 119915P877PC 23.12.2024 Numab Therapeutics AG - Test compound concentration: 1, 10 and 100 ⁇ g/ml - Positive controls: PHA, ⁇ CD3, ⁇ CD52; applied in concentrations of 10 and 100 ⁇ g/ml - Read outs: 1) Immune cell count (CD45+, TCRa/b+, CD4 T cells, CD8 T cells, B cells, NK cells, CD14+ monocytes); 2) Immune cell activation (expression of CD69 & CD25 on T cells); 3) Human cytokine release: CD8/NK cell Panel (Legendplex) - IL-17A, IL-2, IL-4, IL-10, IL-6, TNF- ⁇ , Fas, FasL, IFN- ⁇ , Granzyme A, Granzyme B, Perforin, Granulys
  • PRO5052 and PRO5055 show good safety profile in human whole blood. Their overall cytokine release is minimal and magnitudes lower than positive controls PHA, ⁇ CD52, coated ⁇ CD3 and soluble ⁇ CD3.
  • PRO5052 and PRO5055 show similar or even lower cytokine release than the clinically tested PRO3599 (Pfizer’s anti-PDL1 x CD47 mAB), which showed good safety profile in clinical studies.
  • PRO5053, PRO5054, PRO5605, PRO5423, PRO5056, PRO5057, PRO5606 and PRO5134 were virtually identical to PRO5052 and PRO5055 and are thus not shown.
  • the results for immune cell counts and immune cell activation are summarized in Table 46.
  • PRO5052 and PRO5055 did not induce immune cell activation or changes in immune cell count, while the positive controls showed expected results: PHA induced depletion of human immune cells, ⁇ CD52 induced depletion of immune cells such as T cells and B cells, but not monocytes.
  • Activation (expression of CD69) of remaining T cells in PHA and ⁇ CD52 treated samples was increased.
  • CD25 was increased by PHA but decreased by ⁇ CD52.

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Abstract

The present invention relates to a multispecific antibody comprising one binding domain, which specifically binds to CD47 (CD47-BD), one binding domain, which specifically binds to PDL1 (PDL1-BD), and optionally one or two further binding domains having specificity for an antigen different from CD47 and PDL1. The present invention further relates to nucleic acid sequences encoding said multispecific antibody, vector(s) comprising said nucleic acid sequences, host cell(s) comprising said nucleic acid sequences or said vector(s), and a method of producing said multispecific antibody. Additionally, the present invention relates to pharmaceutical compositions comprising said multispecific antibody and methods of use thereof.

Description

119915P877PC 23.12.2024 Numab Therapeutics AG MULTISPECIFIC ANTIBODIES HAVING SPECIFICITY FOR CD47 AND PDL1 FIELD OF THE INVENTION [0001] The present invention relates to a multispecific antibody comprising one binding domain, which specifically binds to CD47 (CD47-BD), one binding domain, which specifically binds to PDL1 (PDL1-BD), and optionally one or two further binding domains having specificity for an antigen different from CD47 and PDL1. The present invention further relates to nucleic acid sequences encoding said multispecific antibody, vector(s) comprising said nucleic acid sequences, host cell(s) comprising said nucleic acid sequences or said vector(s), and a method of producing said multispecific antibody. Additionally, the present invention relates to pharmaceutical compositions comprising said multispecific antibody and methods of use thereof. BACKGROUND OF THE INVENTION [0002] CD47 is a 50 kDa membrane receptor belonging to the immunoglobulin superfamily. CD47 consists of an extracellular N-terminal IgV domain, five transmembrane domains, and a short C-terminal intracellular tail. CD47 interacts with membrane integrins, the ligand thrombospondin-1 (TSP-1) and signal-regulatory protein alpha (SIRPα), an innate immune checkpoint protein, which is expressed on myeloid cells such as macrophages and dendritic cells (DCs) to suppress their activity. CD47 is found to be overexpressed in many different tumor types (Sick E, et al., Br. J. Pharmacol., 167 (7): 1415–1430 (2012); Chao MP, et al., Curr. Opin. Immunol, 24 (2): 225–232 (2012)) and has been correlated with poor survival in patients. Through the interaction with SIRPα, CD47 provides a so called “don't eat me signal” to macrophages and the dendritic cells (DCs) of the immune system, enabling CD47 expressing tumor cells to escape the innate immune surveillance. [0003] Therefore, CD47 represents an interesting target in cancer therapy. However, CD47 is also ubiquitously expressed on healthy cells such as red blood cells (RBCs) and platelets in peripheral blood. Blockade of the CD47-SIRPα interaction by monoclonal antibodies has been shown to deplete those cells, often resulting in dose limiting toxicities. [0004] PDL1 (CD274, B7-H1) is a 40 kDa type I transmembrane protein. PDL1 is a surface glycoprotein ligand for PD-1, a key immune checkpoint receptor expressed by activated T and B cells, and mediates immunosuppression. PDL1 is found on both antigen-presenting cells and human cancer cells, such as squamous cell carcinoma of the head and neck, melanoma, and brain tumor, thyroid, thymus, esophagus, lung, breast, gastrointestinal tract, colorectum, liver, pancreas, kidney, adrenal cortex, bladder, urothelium, ovary, and skin 119915P877PC 23.12.2024 Numab Therapeutics AG (Katsuya Y, et al., Lung Cancer.88(2):154–159 (2015); Nakanishi J, et al., Cancer Immunol Immunother.56(8):1173–1182 (2007); Nomi T, et al., Clin Cancer Res.13(7):2151–2157 (2007); Fay AP, et al., J Immunother Cancer.3:3 (2015); Strome SE, et al., Cancer Res. 63(19):6501–6505 (2003); Jacobs JF, et al. Neuro Oncol.11(4):394–402 (2009); Wilmotte R, et al. Neuroreport.16(10):1081–1085 (2005)). PDL1 is rarely expressed on normal tissues but inducibly expressed on tumor site (Dong H, et al., Nat Med.8(8):793–800 (2002); Wang et al., Onco Targets Ther.9: 5023–5039 (2016)). PDL1 downregulates T cell activation and cytokine secretion by binding to PD-1 (Freeman et al., 2000; Latchman et al, 2001). PD-1, activated by PDL1, potentially provides an immune-tolerant environment for tumor development and growth. PDL1 also negatively regulates T-cell function through interaction with another receptor, B7.1 (B7-1, CD80). [0005] Various antibodies against PDL1 have been developed (see, for example, WO 2013/079174 and WO 2017/118321) to be used in the treatment of PDL1 sensitive tumor types, such as those indicated above. However, while encouraging results have been obtained for some patients with these PDL1 sensitive tumor types, there is still a large fraction of patients that do not respond to anti-PDL1 antibody treatment. Besides, many tumor types with low anti-PDL1 sensitivity are also enriched with CD47. [0006] To address at least some of the abovementioned issues, several bispecific antibody therapeutics targeting CD47 and PDL1 have been developed that are currently in pre-clinical state or are tested in early clinical trials. [0007] For example, WO 2021/170082 (A1) discloses anti-CD47xPDL1 bispecific antibodies, wherein either the CD47 targeting moiety or the PDL1 targeting moiety is a full- length antibody, and the other targeting moiety is an antigen-binding fragment (e. g., scFv) linked directly or via a peptide linker to the N- or/and the C-terminus of the full-length antibody. WO 2022/235127 (A1) discloses anti-CD47xPDL1 bispecific antibodies comprising an Fc domain, wherein the antigen-binding domain that specifically binds to CD47 is a Fab fragment located in the Fab arms, and the antigen-binding domain that specifically binds to PDL1 is an scFv fragment that is fused via a peptide linker to the C-terminus of the Fc domain. WO 2019/068302 (A1) discloses anti-CD47xPDL1 bispecific antibodies that are monovalent for each specificity, wherein both binding sites are located in the Fab arms and one Fab arm is replaced by an scFv or VHH fragment. It was found that this triple-chain asymmetric antibody format facilitates the separation of the desired bispecific molecule from unwanted immunoglobulin chain combinations. WO 2021/124073 (A1) discloses IgG-based bispecific heterodimeric antibodies having one CD47-binding domain and one PDL1-binding domain that are located in the Fab arms. One of these bispecific antibodies, i. e. PF- 07257876, is currently tested in phase I clinical trials. WO 2019/129054 (A1) discloses IgG- like, Fc-silenced triple-chain antibodies having one CD47-binding domain located in one Fab 119915P877PC 23.12.2024 Numab Therapeutics AG arm and two VHH PDL1-binding domains that are replacing the second Fab arm. One of these bispecific antibodies, i. e. IBI-322, is currently tested in phase II clinical trials. [0008] It is generally expected that these anti-CD47xPDL1 bispecific antibodies have greater selectivity towards PDL1- and CD47-expressing tumor cells and/or PDL1- and CD47-expressing immune cells in a tumor microenvironment than anti-CD47 monospecific antibodies. However, while most of these bispecific antibodies are reported to exhibit reduced or no binding to CD47 on red blood cells, adverse effects caused by red blood cell agglutination are still observed in clinical trials. Some of these bispecific antibodies also suffer from suboptimal efficacy. In this regard, the effect of the Fc region on the efficacy of these bispecific antibodies appears to be unclear, as the presence of active or silenced Fc does not correlate well with the observed clinical efficacy. [0009] There is thus a need in the field to generate improved antibody therapeutics targeting CD47 and PDL1. [0010] Said antibody therapeutics should have a high potency against CD47- and PDL1- expressing cancer cells and at the same time a tolerable safety profile to keep dose limiting side effects, such as in particular red blood cell agglutination, at a low level. Furthermore, it is desirable that said therapeutic antibodies have superior biophysical properties, such as a high stability, in order to facilitate developability and producibility in high yields. SUMMARY OF THE INVENTION [0011] It is an object of the present invention to provide novel and improved therapeutic antibodies that can efficiently eliminate CD47- and PDL1-expressing cancer cells while exhibiting a low level of dose limiting side effects, in particular a low level of red blood cell agglutination. It is a further object of the present invention that said novel and improved therapeutic antibodies exhibit high stability to facilitate developability and producibility in high yields. [0012] The inventors have now surprisingly found that multispecific antibodies having one highly affine PDL1-BD and one moderately affine CD47-BD, in combination with a well- defined IgG-like antibody architecture, wherein the CD47-BD is present in the form of an scFv, exhibit efficient killing of CD47- and PDL1-expressing cancer cells, while causing virtually no agglutination or phagocytosis of red blood cells, even at higher nanomolar concentrations. This was particularly unexpected, since alternative antibody architectures, e. g. antibody architectures wherein said CD47-BD is not in scFv format and/or the PDL1-BD and the CD47-BD are not located next to each other in the Fab arms, do not exhibit the desired combined properties, i. e. potent cancer cell lysis and at the same time low or no agglutination/lysis of red blood cells. 119915P877PC 23.12.2024 Numab Therapeutics AG [0013] It has further been found that the distinct architecture of these multispecific antibodies allows for the addition of further binding domains at the C-terminus of the Fc region, which enables the incorporation of additional functionalities, such as the blockage of other escape mechanisms or activation of additional immune response pathways, for example by adding LILRB2-BDs to foster further activation of immune cells, such as the activation of macrophages and proliferative activation of T-cells. [0014] Said multispecific antibodies also exhibit very advantageous biophysical properties, in particular an outstanding stability, which allows high yield production and the preparation of storage stable formulations. For example, the loss of monomeric content for PRO4672, PRO5052, PRO4687 and PRO5055, which are representative multispecific antibodies of the present invention, is less than 1 % when stored for two months at 4°C at a concentration of 10 mg/ml in a 20 mM histidine buffer (pH 6.0) with 125 mM NaCl and 0.02 % Poloxamer 188. This high stability is particularly surprising in view of the fact that the multispecific antibodies of the invention comprise up to four scFv antibody fragments. [0015] Accordingly, in a first aspect, the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequence of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequence of SEQ ID NO: 7; or ii. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequence of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequence of SEQ ID NO: 7. b) one binding domain, which specifically binds to PDL1 (PDL1-BD); and c) an IgG-like region comprising an immunoglobulin Fc region, a first Fab arm and a second Fab arm, wherein: - the CD47 BD is an scFv fragment replacing the first Fab arm, and - the PDL1 BD is the binding domain of the second Fab arm or is an scFv fragment replacing said second Fab arm. [0016] In a second aspect, the present invention relates to a nucleic acid or two nucleic acids or three nucleic acids encoding the multispecific antibody of the present invention. 119915P877PC 23.12.2024 Numab Therapeutics AG [0017] In a third aspect, the present invention relates to a vector or two vectors or three vectors comprising the nucleic acid or two nucleic acids or three nucleic acids. [0018] In a fourth aspect, the present invention relates to a host cell or host cells comprising the vector or the two vectors or the three vectors of the present invention. [0019] In a fifth aspect, the present invention relates to a method for producing the multispecific antibody of the present invention, comprising (i) providing the nucleic acid or two nucleic acids or three nucleic acids of the present invention, or the vector or the two vectors or the three vectors of the present invention, expressing said nucleic acid or nucleic acids, or said vector or vectors, and collecting said multispecific antibody from the expression system, or (ii) providing a host cell or host cells of the present invention, culturing said host cell or said host cells; and collecting said multispecific antibody from the cell culture. [0020] In a sixth aspect, the present invention relates to a pharmaceutical composition comprising the multispecific antibody of the present invention and a pharmaceutically acceptable carrier. [0021] In a seventh aspect, the present invention relates to a pharmaceutical composition comprising: a) a first multispecific antibody that specifically binds to CD47 and PDL1; b) a second antibody that specifically binds to LILRB2; and c) a pharmaceutically acceptable carrier. [0022] In an eighth aspect, the present invention relates to a multispecific antibody of the present invention for use as a medicament. [0023] In a ninth aspect, the present invention relates to a multispecific antibody of the present invention for use in the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer. [0024] In a tenth aspect, the present invention relates to a method for the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer, comprising the step of administering the multispecific antibody of the present invention or the pharmaceutical composition of the present invention to a patient in need thereof. [0025] In an eleventh aspect, the present invention relates an antibody binding domain, having specificity for CD47 (CD47-BD), as defined above, wherein said CD47-BD is in a format selected from an Fv, a dsFv, an scFv and a dsscFv, particularly from an Fv and an scFv, particularly wherein said CD47-BD is in the form of an scFv. [0026] The aspects, advantageous features and preferred embodiments of the present invention summarized in the following items, respectively alone or in combination, further contribute to solving the object of the invention: 119915P877PC 23.12.2024 Numab Therapeutics AG 1. A multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD), wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequence of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequence of SEQ ID NO: 7; or ii. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequence of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequence of SEQ ID NO: 7; b) one binding domain, which specifically binds to PDL1 (PDL1-BD); and c) an IgG-like region comprising an immunoglobulin Fc region, a first Fab arm and a second Fab arm, wherein: - the CD47 BD is an scFv fragment replacing the first Fab arm, and - the PDL1 BD is in the binding domain of the second Fab arm or is an scFv fragment replacing said second Fab arm. 2. The multispecific antibody of item 1, wherein said CD47-BD comprises: the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequence of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequence of SEQ ID NO: 7. 3. The multispecific antibody of item 1 or 2, wherein said PDL1-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequence of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 79, and the LCDR3 sequence of SEQ ID NO: 80; or ii. the HCDR1 sequence of SEQ ID NO: 74, 119915P877PC 23.12.2024 Numab Therapeutics AG the HCDR2 sequence of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 78, and the LCDR3 sequence of SEQ ID NO: 80; or iii. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequence of SEQ ID NO: 21, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequence of SEQ ID NO: 27; or iv. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequence of SEQ ID NO: 21, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequence of SEQ ID NO: 27; or v. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequence of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequence of SEQ ID NO: 27; or vi. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequence of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequence of SEQ ID NO: 27. 4. The multispecific antibody of items 1 or 2, wherein said PDL1-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequence of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 79, and the LCDR3 sequence of SEQ ID NO: 80; or 119915P877PC 23.12.2024 Numab Therapeutics AG ii. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequence of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 78, and the LCDR3 sequence of SEQ ID NO: 80. 5. The multispecific antibody of any of the preceding items, wherein said CD47-BD, when in scFv format, binds to human CD47 with a monovalent dissociation constant (KD) of 1 to 100 nM, particularly with a KD of 2 to 50 nM, particularly of 5 to 25 nM, as measured by surface plasmon resonance (SPR). 6. The multispecific antibody of any of the preceding items, wherein said CD47-BD: a. when in scFv format, has a melting temperature (Tm), determined by differential scanning fluorimetry, in the range of 50°C to 70°C, particularly in the range of 55°C to 65°C, in particular wherein said antibody is formulated in 50 mM phosphate- citrate buffer (NaCiP) at pH 6.4, 150 mM NaCl; b. when in scFv format, has a loss in monomer content, after storage for four weeks at ‑80°C, of less than 3 %, less than 2 %, less than 1 %, particularly less than 0.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer (NaCiP) with 150 mM NaCl at pH 6.4; c. when in scFv format, has a loss in monomer content, after storage for four weeks at 4°C, of less than 3 %, less than 2 %, less than 1 %, particularly less than 0.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer (NaCiP) with 150 mM NaCl at pH 6.4; and/or d. when in scFv format, has a loss in monomer content, after storage for four weeks at 40°C, of less than 3 %, less than 2 %, less than 1 %, particularly less than 0.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer (NaCiP) with 150 mM NaCl at pH 6.4. 7. The multispecific antibody of any of the preceding items, wherein said PDL1-BD: a. when in scFv format, binds to human PDL1 with a monovalent dissociation constant (KD) of 1 to 100 pM, particularly with a KD of 1 to 50 pM, particularly of 1 to 40 pM, as measured by surface plasmon resonance (SPR); 119915P877PC 23.12.2024 Numab Therapeutics AG b. when in scFv format, is cross-reactive with Macaca fascicularis (cynomolgus) PDL1, in particular binds to cynomolgus PDL1 with a monovalent KD of 1 to 100 pM, particularly with a KD of 1 to 50 pM, particularly of 1 to 40 pM, as measured by SPR; c. when in scFv format, has the ability to neutralize PDL1/PD-1 interaction, as measured in an NFAT reporter gene assay, with an EC50 value of 0.1 to 10 nM, particularly of 0.1 to 5 pM, particularly of 0.1 to 3 nM; d. is non-cross-reactive to Mus musculus PDL1, as measured by SPR; and/or e. does not bind to human PDL2, as measured by SPR. 8. The multispecific antibody of any of the preceding items, wherein said PDL1-BD: a. when in scFv format, has a melting temperature (Tm), determined by differential scanning fluorimetry, in the range of 60°C to 85°C, particularly in the range of 65°C to 85°C, particularly in the range of 70°C to 85°C, in particular wherein said antibody is formulated in 50 mM phosphate-citrate buffer at pH 6.4, 150 mM NaCl; b. when in scFv format, has a loss in monomer content, after five consecutive freeze- thaw cycles, of less than 5 %, preferably less than 3 %, more preferably less than 1 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein the antibody of the invention is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4; and/or c. when in scFv format, has a loss in monomer content, after storage for four weeks at 4°C, of less than 15 %, e. g. less than 12 %, less than 10 %, less than 7 %, less than 5 %, less than 4 %, less than 3 %, less than 2 %, preferably less than 1 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4. 9. The multispecific antibody of any of the preceding items, wherein the KD of the PDL1- BD of said multispecific antibody for binding to human PDL1 is at least 500 fold lower, in particular at least 1000 fold lower, in particular at least 1500 fold lower, in particular at least 2000 fold lower, than the KD of the CD47 BD of said multispecific antibody for binding to human CD47. 10. The multispecific antibody of any of the preceding items, wherein said multispecific antibody does not trigger any visible red blood cell agglutination at a concentration of 5 nM, particularly of 20 nM, particularly of 50 nM, particularly of 100 nM, particularly of 200 nM, particularly of 500 nM, as determined in a standard hemagglutination assay as 119915P877PC 23.12.2024 Numab Therapeutics AG described in Example 4.2; and/or does not cause the lysis of red blood cells at a concentration of 5 nM, particularly of 20 nM, particularly of 50 nM, as determined in a standard phagocytosis assay, as described in Example 5.4. 11. The multispecific antibody of item 1, wherein the immunoglobulin Fc region is selected from an immunoglobulin Fc region of an IgG subclass, particularly an immunoglobulin Fc region of human lgG1, lgG2 (lgG2A, IgG2B), lgG3 or lgG4, particularly of human IgG1 or IgG2, particularly of human IgG1. 12. The multispecific antibody of item 1, wherein the immunoglobulin Fc region is an active Fc region. 13. The multispecific antibody of any one of the preceding items, wherein the immunoglobulin Fc region has one or more amino acid modifications selected from the group consisting of: a) modifications to enhance heterodimerization of heavy chains, such as the knob- into-hole (KiH) technology; b) modifications that increase or decrease Fc receptor-binding, when compared to the respective unmodified immunoglobulin Fc region thereof; c) modifications that increase serum half-life, when compared to the respective unmodified immunoglobulin Fc region thereof. 14. The multispecific antibody of any one of the preceding items, wherein said CD47-BD comprises VH3 or VH4 domain framework sequences FR1 to FR4; particularly VH3 domain framework sequences FR1 to FR4. 15. The multispecific antibody of item 14, wherein said CD47-BD comprises a VL domain comprising a VL framework comprising framework regions FR1, FR2 and FR3, which are selected from Vκ subtypes, particularly from the Vκ1 and Vκ3 subtypes, particularly are of the Vκ1 subtype, and a framework FR4, which is a Vλ FR4, particularly is a Vλ FR4 comprising an amino acid sequence having at least 80, 90 percent identity to any of SEQ ID NOs: 161 to 169, more particularly a Vλ FR4 selected from any of SEQ ID NOs: 161 to 169, particularly a Vλ FR4 according to SEQ ID NO: 161 or 168. 16. The multispecific antibody of any one of the preceding items, wherein said CD47-BD comprises a) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 8 and 9; and 119915P877PC 23.12.2024 Numab Therapeutics AG a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 11 and 12, particularly to SEQ ID NO: 12; or b) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 10; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 13. 17. The multispecific antibody of any one of the preceding items, wherein said CD47-BD comprises a) a VH sequence selected from any one of SEQ ID NOs: 8 and 9; and a VL sequence selected from any one of SEQ ID NOs: 11 and 12, particularly the VL sequence of SEQ ID NO: 12; or b) the VH sequence of SEQ ID NO: 10; and the VL sequence of SEQ ID NO: 13. 18. The multispecific antibody of any one of the preceding items, wherein said PDL1-BD comprises VH3 or VH4 domain framework sequences FR1 to FR4; particularly VH3 domain framework sequences FR1 to FR4. 19. The multispecific antibody of item 18, wherein said PDL1-BD comprises a) a VL domain comprising a VL framework comprising framework regions FR1, FR2, FR3 and FR4, which are selected from Vκ subtypes, particularly from the Vκ1 and Vκ3 subtypes, particularly are of the Vκ1 subtype; or b) a VL domain comprising a VL framework comprising framework regions FR1, FR2 and FR3, which are selected from Vκ subtypes, particularly from the Vκ1 and Vκ3 subtypes, particularly are of the Vκ1 subtype, and a framework FR4, which is selected from a Vλ FR4, particularly is a Vλ FR4 comprising an amino acid sequence having at least 80, 90 percent identity to any of SEQ ID NOs: 161 to 169, more particularly a Vλ FR4 selected from any of SEQ ID NOs: 161 to 169, particularly a Vλ FR4 according to SEQ ID NO: 161 or 168. 20. The multispecific antibody of any one of the preceding items, wherein said PDL1-BD comprises a) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 28; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 34, 36 and 37, particularly selected from SEQ ID NOs: 34 and 37, particularly to SEQ ID NO:37; or b) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 29 and 31; and 119915P877PC 23.12.2024 Numab Therapeutics AG a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 33 and 35, particularly to SEQ ID NO: 35; or c) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 30; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 32 and 34, particularly to SEQ ID NO: 34; or d) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 38 and 42; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 48, 50 and 51, particularly selected from SEQ ID NOs: 48 and 51, particularly to SEQ ID NO: 51; or e) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 39, 41, 43 and 45; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 47 and 49, particularly to SEQ ID NO: 49; or f) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 40 and 44; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 46 and 48, particularly to SEQ ID NO: 48; or g) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 52; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 58, 60 and 61, particularly selected from SEQ ID NOs: 58 and 61, particularly to SEQ ID NO: 61; or h) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 53 and 55; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 57 and 59, particularly to SEQ ID NO: 59; or i) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 54; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 56 and 58, particularly to SEQ ID NO: 58; or j) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 81; and 119915P877PC 23.12.2024 Numab Therapeutics AG a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 86, 88 and 89, particularly selected from SEQ ID NOs: 86 and 89, particularly to SEQ ID NO: 89; or k) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 83; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 85 and 86, particularly to SEQ ID NO: 86; or l) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 82 and 84, particularly to SEQ ID NO: 84; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequences of SEQ ID NO: 86. 21. The multispecific antibody of any one of the preceding items, wherein said PDL1-BD comprises a) the VH sequence of SEQ ID NO: 28; and a VL sequence selected from any one of SEQ ID NOs: 34, 36 and 37, particularly selected from any one of SEQ ID NOs: 34 and 37, particularly the VL sequence of SEQ ID NO: 37; or b) a VH sequence selected from any one of SEQ ID NOs: 29 and 31; and a VL sequence selected from any one of SEQ ID NOs: 33 and 35, particularly the VL sequence of SEQ ID NO: 35; or c) the VH sequence of SEQ ID NO: 30; and a VL sequence selected from any one of SEQ ID NOs: 32 and 34, particularly the VL sequence of SEQ ID NO: 34; or d) a VH sequence selected from any one of SEQ ID NOs: 38 and 42; and a VL sequence selected from any one of SEQ ID NOs: 48, 50 and 51, particularly from SEQ ID NOs: 48 and 51, particularly the VL sequence of SEQ ID NO: 51; or e) a VH sequence selected from any one of SEQ ID NOs: 39, 41, 43 and 45; and a VL sequence selected from any one of SEQ ID NOs: 47 and 49, particularly the VL sequence of SEQ ID NO: 49; or f) a VH sequence selected from any one of SEQ ID NOs: 40 and 44; and a VL sequence selected from any one of SEQ ID NOs: 46 and 48, particularly the VL sequence of SEQ ID NO: 48; or g) the VH sequence of SEQ ID NO: 52; and a VL sequence selected from any one of SEQ ID NOs: 58, 60 and 61, particularly from SEQ ID NOs: 58 and 61, particularly the VL sequence of SEQ ID NO:61; or h) a VH sequence selected from any one of SEQ ID NO: 53 and 55; and a VL sequence selected from any one of SEQ ID NOs: 57 and 59, particularly the VL sequence of SEQ ID NO: 59; or i) the VH sequence of SEQ ID NO: 54; and 119915P877PC 23.12.2024 Numab Therapeutics AG a VL sequence selected from any one of SEQ ID NOs: 56 and 58, particularly the VL sequence of SEQ ID NO: 58; or j) the VH sequence of SEQ ID NO: 81; and a VL sequence selected from any one of SEQ ID NOs: 86, 88 and 89, particularly from SEQ ID NOs: 86 and 89, particularly the VL sequence of SEQ ID NO: 89; or k) the VH sequence of SEQ ID NO: 83; and a VL sequence selected from any one of SEQ ID NOs: 85 and 86, particularly the VL sequence of SEQ ID NO: 86; or l) a VH sequence selected from any one of SEQ ID NOs: 82 and 84, particularly the VH sequence of SEQ ID NO: 84; and the VL sequence of SEQ ID NO: 86. 22. The multispecific antibody of any one of the preceding items, wherein said PDL1-BDs comprises CDR regions derived from a parental rabbit antibody. 23. The multispecific antibody of any one of the preceding items, wherein said CD47-BD comprises: a) a VH sequence selected from any one of SEQ ID NOs: 8 and 9; and the VL sequence of SEQ ID NO: 12; or b) the VH sequence of SEQ ID NO: 10; and the VL sequence of SEQ ID NO: 13; and wherein said PDL1-BD comprises: a) the VH sequence of SEQ ID NO: 52; and a VL sequence selected from any one of SEQ ID NOs: 58 and 61, particularly the VL sequence of SEQ ID NO:61; or b) the VH sequence of SEQ ID NO: 54; and the VL sequence of SEQ ID NO: 58; or d) the VH sequence of SEQ ID NO: 81; and a VL sequence selected from any one of SEQ ID NOs: 86 and 89, particularly the VL sequence of SEQ ID NO: 89; or e) the VH sequence of SEQ ID NO: 83; and the VL sequence of SEQ ID NO: 86. 24. The multispecific antibody of any one of the preceding items, which has amino acid sequences selected from: - SEQ ID NO: 174 and SEQ ID NO: 175; - SEQ ID NO: 176 SEQ ID NO: 177 and SEQ ID NO: 178; - SEQ ID NO: 179, SEQ ID NO: 180 and SEQ ID NO: 181; - SEQ ID NO: 182 and SEQ ID NO: 183; - SEQ ID NO: 184, SEQ ID NO: 185 and SEQ ID NO: 186; 119915P877PC 23.12.2024 Numab Therapeutics AG - SEQ ID NO: 197 and SEQ ID NO: 198; - SEQ ID NO: 199, SEQ ID NO: 200 and SEQ ID NO: 201; - SEQ ID NO: 202 and SEQ ID NO: 203; and - SEQ ID NO: 204, SEQ ID NO: 205 and SEQ ID NO: 206. 25. The multispecific antibody of any one of the preceding items, wherein said multispecific antibody further comprises: d) one further binding domain, which has specificity for an antigen different from CD47 and PDL1, wherein said one further binding domain is attached to one of the C-termini of the heavy chains of the immunoglobulin Fc region. 26. The multispecific antibody of any one of the items 1 to 24, wherein the multispecific antibody further comprises: d) two further binding domains, which have specificity for an antigen different from CD47 and PDL1, wherein said two further binding domains are attached to each of the C-termini of the heavy chains of the immunoglobulin Fc region. 27. The multispecific antibody of item 25 or 26, wherein the one or two further binding domains specifically bind to a tumor associated antigen (TAA) different from CD47 and PDL1. 28. The multispecific antibody of item 27, wherein the TAA is selected from ADRB3, AFP, ALK, BCMA, beta human chorionic gonadotropin, CA-125 (MUC16), CAIX, CD123, CD133, CD135, CD135 (FLT3), CD138, CD171, CD19, CD20, CD22, CD24, CD276, CD33, CD33, CD38, CD44v6, CD79b, CD97, CDH3 (cadherin 3), CEA, CEACAM6, CLDN6, CLEC12A (CLL1), CSPG4, CYP1B1, EGFR, EGFRvlll, EPCAM, EPHA2, Ephrin B2, ERBBs (e. g. ERBB2), FAP, FGFR1, folate receptor alpha, folate receptor beta, Fos-related antigen, GA733, GD2, GD3, GFRalpha4, globoH, GPC3, GPR20, GPRC5D, HAVCR1, Her2/neu (HER2), HLA-A2, HMWMAA, HPV E6 or E7, human telomerase reverse transcriptase, IL-11Ra, IL-13Ra2, intestinal carboxyl esterase, KIT, Legumain, LewisY, LMP2, Ly6k, MAD-CT-1, MAD-CT-2, ML-IAP, MN-CA IX, MSLN, MUC1, mut hsp 70-2, NA-17, NCAM, neutrophil elastase, NY-BR-1, NY-ESO- 1, o-acetyl-GD2, OR51E2, PANX3, PDGFR-beta, PLAC1, Polysialic acid, PSCA, PSMA, RAGE1, ROR1, sLe, sperm protein 17, SSEA-4, SSTR2, sTn antigen, sTn-O- Glycopeptides, TAG72, TARP, TEM1/CD248, TEM7R, thyroglobulin, Tn antigen, Tn-O- Glycopeptides, TPBG (5T4), TRP-2, TSHR, UPK2 and VEGFR2. 119915P877PC 23.12.2024 Numab Therapeutics AG 29. The multispecific antibody of item 25 or 26, wherein the one or two further binding domains specifically bind to an NK cell receptor and/or an immunomodulatory antigen different from PDL1. 30. The multispecific antibody of any one of items 25, 26 and 29, wherein at least one of the further binding domains specifically binds to an NK cell receptor. 31. The multispecific antibody of item 29 or 30, wherein the immunomodulatory antigen different from PDL1 is selected from B7-1 (CD80), B7-2 (CD86), B7-DC (CD273), B7-H1 (CD274), B7-H2 (CD275), B7-H3 (CD276), B7-H4 (VTCN1), B7-H5 (VISTA), BTLA (CD272), 4-1BB (CD137), CD137L, CD24, CD27, CD28, CD38, CD40, CD40L (CD154), CD54, CD59, CD70, CTLA-1, CTLA-4 (CD152), CXCL9, GITR (CD357), GITR, HVEM (CD270), ICAM-1 (CD54), ICOS (CD278), LAG-3 (CD223), OX40 (CD134), OX40L (CD252), PD-1 (CD279), TIGIT, CD314, CD334, CD335, CD337, VISTA and TIM-3 (CD366). 32. The multispecific antibody of item 26, wherein both of the further binding domains specifically bind to an NK cell receptor. 33. The multispecific antibody of any one of the items 29 to 32, wherein the NK cell receptor is selected from KIR2DL1, KIR2DL2/3, KIR2DL4, KIR2DL5A, KIR2DL5B, KIR3DL1, KIR3DL2, KIR3DL3, KIR2DS4, KIR3DS, NKG2A, NKG2C, NKG2D, NKG2E, NKp30, NKp44, NKp46, CD2, CD16, CD69, DNAX accessory molecule-1 (DNAM-1), 2B4, NK1.1; a killer immunoglobulin (Ig)-like activating receptors (KAR); ILTs/LIRs (CD85) such as LILRA1, LILRA2, LILRA3, LILRA4, LILRA5, LILRA6, LILRB1, LILRB2, LILRB3, LILRB4, LILRB5, ILT8, ILT10, ILT11; NKRP-1; CD94/NKG2A, CD94/NKG2C and CD94/NKG2E heterodimers. 34. The multispecific antibody of any one of the items 29 to 32, wherein the NK cell receptor is selected from NKp46, LILRB1, LILRB2, LILRB3, LILRB4 and LILRB5, particularly LILRB1, LILRB2, and LILRB4, particularly LILRB1 and LILRB2, particularly LILRB2. 35. The multispecific antibody of item 25, wherein said further binding domain specifically binds to LILRB2 (LILRB2-BD). 36. The multispecific antibody of item 25, wherein said two further binding domains specifically bind to LILRB2 (LILRB2-BDs). 119915P877PC 23.12.2024 Numab Therapeutics AG 37. The multispecific antibody of item 36, wherein the LILRB2-BDs bind to different epitopes on the extracellular domain of LILRB2. 38. The multispecific antibody of item 36, wherein the LILRB2-BDs bind to the same epitope on the extracellular domain of LILRB2. 39. The multispecific antibody of item 36, wherein the LILRB2-BDs are identical. 40. The multispecific antibody of any one of the items 35 to 39, wherein said LILRB2-BD or said LILRB2-BDs comprise: i. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequence of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 98, and the LCDR3 sequence of SEQ ID NO: 99; or ii. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequence of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 97, and the LCDR3 sequence of SEQ ID NO: 99; or iii. the HCDR1 sequence of SEQ ID NO: 112, the HCDR2 sequence of SEQ ID NO: 113, the HCDR3 sequence of SEQ ID NO: 114, the LCDR1 sequence of SEQ ID NO: 115, the LCDR2 sequence of SEQ ID NO: 117, and the LCDR3 sequence of SEQ ID NO: 118; or iv. the HCDR1 sequence of SEQ ID NO: 112, the HCDR2 sequence of SEQ ID NO: 113, the HCDR3 sequence of SEQ ID NO: 114, the LCDR1 sequence of SEQ ID NO: 115, the LCDR2 sequence of SEQ ID NO: 116, and the LCDR3 sequence of SEQ ID NO: 118 or v. the HCDR1 sequence of SEQ ID NO: 128, the HCDR2 sequence of SEQ ID NO: 129, the HCDR3 sequence of SEQ ID NO: 130, 119915P877PC 23.12.2024 Numab Therapeutics AG the LCDR1 sequence of SEQ ID NO: 131, the LCDR2 sequence of SEQ ID NO: 133, and the LCDR3 sequence of SEQ ID NO: 134; and/or vi. the HCDR1 sequence of SEQ ID NO: 128, the HCDR2 sequence of SEQ ID NO: 129, the HCDR3 sequence of SEQ ID NO: 130, the LCDR1 sequence of SEQ ID NO: 131, the LCDR2 sequence of SEQ ID NO: 132, and the LCDR3 sequence of SEQ ID NO: 134. 41. The multispecific antibody of any one of the items 35 to 39, wherein said LILRB2-BD or said LILRB2-BDs comprise: i. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequence of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 98, and the LCDR3 sequence of SEQ ID NO: 99; or ii. the HCDR1 sequence of SEQ ID NO: 112, the HCDR2 sequence of SEQ ID NO: 113, the HCDR3 sequence of SEQ ID NO: 114, the LCDR1 sequence of SEQ ID NO: 115, the LCDR2 sequence of SEQ ID NO: 117, and the LCDR3 sequence of SEQ ID NO: 118 and/or iii. the HCDR1 sequence of SEQ ID NO: 128, the HCDR2 sequence of SEQ ID NO: 129, the HCDR3 sequence of SEQ ID NO: 130, the LCDR1 sequence of SEQ ID NO: 131, the LCDR2 sequence of SEQ ID NO: 133, and the LCDR3 sequence of SEQ ID NO: 134. 42. The multispecific antibody of any one of the items 35 to 39, wherein said LILRB2-BD or said LILRB2-BDs comprise: i. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequence of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, 119915P877PC 23.12.2024 Numab Therapeutics AG the LCDR2 sequence of SEQ ID NO: 98, and the LCDR3 sequence of SEQ ID NO: 99; and/or ii. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequence of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 97, and the LCDR3 sequence of SEQ ID NO: 99. 43. The multispecific antibody of any one of the items 35 to 39, wherein said LILRB2-BD or both said LILRB2-BDs comprise: i. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequence of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 98, and the LCDR3 sequence of SEQ ID NO: 99. 44. The multispecific antibody of any one of the items 35 to 43, wherein said LILRB2-BD or said LILRB2-BDs: a. when in scFv format, bind to human LILRB2 with a monovalent dissociation constant (KD) of 10 pM to 5 nM, particularly with a KD of 20 pM to 3 nM, particularly of 50 pM to 1 nM, as measured by surface plasmon resonance (SPR); b. block the interaction of LILRB2 to its ligand HLA-G with an IC50 of 100 pM to 10 nM, particularly of 100 pM to 5 nM, particularly of 100 pM to 2 nM, as measured by flow cytometry; c. block the interaction of LILRB2 to its ligand HLA-A3 with an IC50 of 100 pM to 10 nM, particularly of 100 pM to 5 nM, particularly of 100 pM to 2 nM, as measured by flow cytometry; d. are not cross-reactive with members of leukocyte immunoglobulin-like receptor subfamily A (LILRA); and/or e. are not cross reactive with members of leukocyte immunoglobulin-like receptor subfamily LILRB3, LILRB4 and LILRB5. 45. The multispecific antibody of any one of the items 35 to 44, wherein said LILRB2-BD or said LILRB2-BDs: a. when in scFv format, have a melting temperature (Tm), determined by differential scanning fluorimetry, in the range of 55°C to 85°C, particularly in the range of 60°C 119915P877PC 23.12.2024 Numab Therapeutics AG to 85°C, in particular wherein said antibodies are formulated in 50 mM phosphate- citrate buffer at pH 6.4, 150 mM NaCl; b. when in scFv format, have a loss in monomer content, after storage for four weeks, at ‑80°C, of less than 5 %, less than 4 %, less than 3 %, less than 2 %, particularly less than 1.5 %, when said antibodies are at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4; c. when in scFv format, has a loss in monomer content, after storage for four weeks, at 4°C, of less than 15 %, e. g. less than 12 %, less than 10 %, less than 9 %, less than 8 %, particularly less than 7 %, when said antibodies are at a starting concentration of 10 mg/ml, and in particular wherein said antibodies are formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4; d. when in scFv format, has a loss in monomer content, after storage for four weeks, at 40°C, of less than 15 %, e. g. less than 12 %, less than 10 %, less than 9 %, less than 8 %, particularly less than 7 %, when said antibodies are at a starting concentration of 10 mg/ml, and in particular wherein said antibodies are formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4. 46. The multispecific antibody of any one of the items 35 to 45, wherein said LILRB2-BD or said LILRB2-BDs are in a format independently selected from a Fab, an scFab, an Fv, a dsFv, an scFv and a dsscFv, particularly from a Fab, Fv and scFv. 47. The multispecific antibody of any one of the items 35 to 46, wherein said LILRB2-BD or said LILRB2-BDs comprise VH3 or VH4 domain framework sequences FR1 to FR4; particularly VH3 domain framework sequences FR1 to FR4. 48. The multispecific antibody of any one of the items 35 to 47, wherein said LILRB2-BD or said LILRB2-BDs independently comprise a) a VL domain comprising a VL framework comprising framework regions FR1, FR2, FR3 and FR4, which are selected from Vκ subtypes, particularly from the Vκ1 and Vκ3 subtypes, particularly are of the Vκ1 subtype; or b) a VL domain comprising a VL framework comprising framework regions FR1, FR2 and FR3, which are selected from Vκ subtypes, particularly from the Vκ1 and Vκ3 subtypes, particularly are of the Vκ1 subtype, and a framework FR4, which is selected from a Vλ FR4, particularly is a Vλ FR4 comprising an amino acid sequence having at least 80, 90 percent identity to any of SEQ ID NOs: 161 to 169, more particularly a Vλ FR4 selected from any of SEQ ID NOs: 161 to 169, particularly a Vλ FR4 according to SEQ ID NO: 161 or 168. 119915P877PC 23.12.2024 Numab Therapeutics AG 49. The multispecific antibody of any one of the items 35 to 48, wherein said LILRB2-BD or said LILRB2-BDs independently comprise a) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 100; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 104, 106 and 107, particularly selected from SEQ ID NOs: 104 and 107, particularly to SEQ ID NO: 107; or b) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 101; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 103 and 104, particularly to SEQ ID NO: 104; or c) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 102; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 105; or d) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 119; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 122, 123 and 124, particularly selected from SEQ ID NOs: 122 and 124, particularly to SEQ ID NO: 124; or e) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 120; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 121 and 122, particularly to SEQ ID NO: 122; or f) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 135; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 138, 139 and 140, particularly selected from SEQ ID NOs: 138 and 140, particularly to SEQ ID NO: 140; or g) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 136; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 138. 50. The multispecific antibody of any one of the items 35 to 48, wherein said LILRB2-BD or said LILRB2-BDs independently comprise a) a VH sequence of SEQ ID NO: 100; and 119915P877PC 23.12.2024 Numab Therapeutics AG a VL sequence selected from SEQ ID NOs: 104, 106 and 107, particularly selected from SEQ ID NOs: 104 and 107, particularly from SEQ ID NO: 107; or b) a VH sequence of SEQ ID NO: 101; and a VL sequence selected from SEQ ID NOs: 103 and 104, particularly from SEQ ID NO: 104; or c) a VH sequence of SEQ ID NO: 102; and a VL sequence of SEQ ID NO: 105; or d) a VH sequence of SEQ ID NO: 119; and a VL sequence selected from SEQ ID NOs: 122, 123 and 124, particularly selected from SEQ ID NOs: 122 and 124, particularly from SEQ ID NO: 124; or e) a VH sequence of SEQ ID NO: 120; and a VL sequence selected from SEQ ID NOs: 121 and 122, particularly from SEQ ID NO: 122; or f) a VH sequence of SEQ ID NO: 135; and a VL sequence selected from SEQ ID NOs: 138, 139 and 140, particularly from SEQ ID NOs: 138 and 140, particularly from SEQ ID NO: 140; or g) a VH sequence of SEQ ID NO: 136; and a VL sequence of SEQ ID NO: 138. 51. The multispecific antibody of any one of the items 35 to 50, wherein said multispecific antibody is further characterized by one or more of the following features: a) said LILRB2-BD comprises CDR regions derived from a parental rabbit antibody; b) binds to human CD47 with a monovalent dissociation constant (KD) of 1 to 100 nM, particularly with a KD of 5 to 70 nM, particularly of 10 to 50 nM, as measured by surface plasmon resonance (SPR); c) binds to human PDL1 with a monovalent dissociation constant (KD) of 1 to 200 pM, particularly with a KD of 1 to 150 pM, particularly of 1 to 100 pM, as measured by surface plasmon resonance (SPR); d) binds to human LILRB2 with a monovalent dissociation constant (KD) of 10 pM to 5 nM, particularly with a KD of 20 pM to 3 nM, particularly of 50 pM to 1 nM, as measured by surface plasmon resonance (SPR); e) does not cause the lysis of red blood cells at a concentration of 10 nM, particularly of 50 nM, particularly of 100 nM as determined in a standard phagocytosis assay, as described in Example 5.4; f) blocks the interaction of CD47 to SIRPα with an IC50 of 10 to 500 pM, particularly of 10 to 300 pM, particularly of 10 to 200 pM, as measured by flow cytometry using NCI-H292 cells; g) has a second melting temperature (Tm2), determined by differential scanning fluorimetry, in the range of 60°C to 80°C, particularly in the range of 63°C to 80°C, particularly in the range of 65°C to 80°C, in particular wherein said multispecific antibody is formulated in 20 mM Histidine buffer at pH 6.0, 125 mM NaCl and 0.02 % Poloxamer 188; 119915P877PC 23.12.2024 Numab Therapeutics AG h) has a loss in monomer content, after five consecutive freeze-thaw cycles, of less than 2 %, preferably less than 1 %, more preferably less than 0.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein the antibody of the invention is formulated in 20 mM Histidine buffer at pH 6.0, 125 mM NaCl and 0.02 % Poloxamer 188; i) has a loss in monomer content, after storage for 8 weeks at 4°C, of less than 5 %, e. g. less than 3 %, less than 2 %, less than 1 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 20 mM Histidine buffer at pH 6.0, 125 mM NaCl and 0.02 % Poloxamer 188; j) has a loss in monomer content, after storage for 4 weeks at 25°C, of less than 5 %, e. g. less than 3 %, less than 2 %, less than 1 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 20 mM Histidine buffer at pH 6.0, 125 mM NaCl and 0.02 % Poloxamer 188; and/or k) has a loss in monomer content, after storage for 4 weeks at 40°C, of less than 7 %, e. g. less than 6 %, less than 5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 20 mM Histidine buffer at pH 6.0, 125 mM NaCl and 0.02 % Poloxamer 188. 52. The multispecific antibody of any one of the items 35 to 51, wherein the LILRB2-BD and the CD47-BD are capable of binding simultaneously to their respective antigens. 53. The multispecific antibody of any one of the items 35 to 52, which has amino acid sequences selected from: - SEQ ID NO: 187 and SEQ ID NO: 188 (PRO4672); - SEQ ID NO: 189, SEQ ID NO: 190 and SEQ ID NO: 191 (PRO4687); - SEQ ID NO: 192 and SEQ ID NO: 193 (PRO5052); and - SEQ ID NO: 194, SEQ ID NO: 195 and SEQ ID NO: 196 (PRO5055). 54. A nucleic acid or two nucleic acids or three nucleic acids encoding the multispecific antibody of any one of items 1 to 53. 55. A vector or two vectors or three vectors comprising the nucleic acid or two nucleic acids or three nucleic acids of item 54. 56. A host cell or host cells comprising the vector or the two vectors or the three vectors of item 55. 57. A method for producing the multispecific antibody of any one of items 1 to 53, comprising (i) providing the nucleic acid or two nucleic acids or three nucleic acids of item 54, or the 119915P877PC 23.12.2024 Numab Therapeutics AG vector or the two vectors or the three vectors of item 55, expressing said nucleic acid or nucleic acids, or said vector or vectors, and collecting said multispecific antibody from the expression system, or (ii) providing a host cell or host cells according to item 56, culturing said host cell or said host cells; and collecting said multispecific antibody from the cell culture. 58. A pharmaceutical composition comprising the multispecific antibody of any one of items 1 to 53 and a pharmaceutically acceptable carrier. 59. A pharmaceutical composition comprising: a) a first multispecific antibody that specifically binds to CD47 and PDL1; b) a second antibody that specifically binds to LILRB2; and c) a pharmaceutically acceptable carrier. 60. The pharmaceutical composition of item 59, wherein the first antibody that specifically binds to CD47 and PDL1 is selected from anti-CD47xPDL1 antibodies that are currently tested or have been tested in clinical trials, preferably wherein the first antibody that specifically binds to CD47 and PDL1 is selected from the group consisting of: IBI-322 by Innovent Biologics Inc.; PF-07257876 by Pfizer Inc.; 6MW-3211 by Mabwell (Shanghai) Bioscience Co. Ltd.; BAT7104 by Bio-Thera Solutions, Ltd.; QL401 by QLSF Biotherapeutics, Inc.; LB-101 by Centessa Pharmaceuticals plc; Imm2520 by ImmuneOnco Biopharmaceuticals Shanghai Inc.; SHS-009 by Nanjing Sanhome Pharmaceutical Co Ltd.; BCD-106 by Biocad Biopharmaceutical Co.; TJL-1C4 by I-MAB Biopharma Co. Ltd.; SG12473 by Hangzhou Sumgen Biotech Co. Ltd.; XB014 by Exelixis Inc.; PMC-122 by PharmAbcine Inc.; ABP-160 by Abpro Corp.; papiliximab by Shaperon Inc.; JY-207 by Shenzhen Enduring Biotech Ltd.; NY2601 and NY2901 by Light Chain Bioscience AG.. 61. The pharmaceutical composition of item 59, wherein the first antibody that specifically binds to CD47 and PDL1 is selected from a multispecific antibody as defined in any one of the items 1 to 24. 62. The pharmaceutical composition of any one of the items 59 to 61, wherein the second antibody that specifically binds to LILRB2 is selected from anti-LILRB2 antibodies that are currently tested or have been tested in clinical trials, preferably wherein the second antibody that specifically binds to LILRB2 is selected from the group consisting of: JTX- 8064 by Jounce Therapeutics Inc.; MK-4830 by Merck & Co. Inc.; IO-108 by Immune- Onc Therapeutics Inc.; NGM-707 by NGM Biopharmaceuticals Inc.; BMS-986406 by 119915P877PC 23.12.2024 Numab Therapeutics AG Bristol Myers Squibb Company; IOS-1002 by ImmunOs Therapeutics AG; CDX-585 by Celldex Therapeutics Inc.; SPX-303 by SparX Biopharmaceutical Corp.; CHS-1000 by Coherus BioSciences Inc.; ES009 by Elpiscience (Suzhou) Biopharma Ltd.; XBH41 by GV20 Therapeutics and IVS-5001 by Invectys. 63. The pharmaceutical composition of any one of the items 59 to 61, wherein the second antibody that specifically binds to LILRB2 comprises at least one LILRB2-BD as defined in any one of the items 40 to 50. 64. The pharmaceutical composition of any one of the items 59 to 61 and 63, wherein the second antibody comprises two LILRB2-BDs. 65. The pharmaceutical composition of item 64, wherein the two LILRB2-BDs bind to different epitopes on the extracellular domain of LILRB2. 66. The pharmaceutical composition of item 64, wherein the two LILRB2-BDs bind to the same epitope on the extracellular domain of LILRB2. 67. The pharmaceutical composition of item 66, wherein the LILRB2-BDs are identical. 68. The pharmaceutical composition of any one of the items 63 to 67, wherein the second antibody is a fragment-based antibody further comprising one hSA-binding domain. 69. The pharmaceutical composition of item 68, wherein the format of the second antibody is selected from a tandem scDb (Tandab), a linear dimeric scDb (LD-scDb), a circular dimeric scDb (CD-scDb), a tandem tri-scFv, a tribody (Fab-(scFv)2), a Fab-Fv2, a triabody, an scDb-scFv, a tetrabody, a didiabody, a tandem-di-scFv and a MATCH. 70. The pharmaceutical composition of any one of the items 63 to 67, wherein the second antibody comprises an Fc region, as defined in items 11 and 12. 71. The pharmaceutical composition of item 70, wherein the immunoglobulin Fc region is selected from a) an immunoglobulin Fc region of an IgG subclass, particularly an immunoglobulin Fc region of human lgG1, lgG2 (lgG2A, IgG2B), lgG3 or lgG4, particularly of human IgG1 or IgG4; 119915P877PC 23.12.2024 Numab Therapeutics AG b) an immunoglobulin Fc region of a) having 1, 2 or 3 amino acid modifications, which exhibits increased or decreased Fc receptor-binding when compared to the respective unmodified immunoglobulin Fc region of a). 72. The pharmaceutical composition of item 70 or 71, wherein the format of the second antibody is selected from an IgG format. 73. The multispecific antibody of any one of items 1 to 53 for use as a medicament. 74. The multispecific antibody of any one of items 1 to 53 or the pharmaceutical composition of any one of items 58 to 72 for use in the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer. 75. The multispecific antibody of item 74, wherein the cancer is selected from a benign or especially malignant tumor, solid tumors, mesothelioma, brain cancer, kidney cancer, liver cancer, adrenal gland cancer, bladder cancer, breast cancer, stomach cancer (e. g., gastric tumors), esophageal cancer, ovarian cancer, cervical cancer, thymic cancer, choriocarcinoma, oral cancer, salivary cancer, colon cancer, rectum cancer, prostate cancer, pancreatic cancer, lung cancer (e. g., non-small cell lung cancer and small cell lung cancer), vaginal cancer, thyroid cancer, melanoma (e. g., unresectable or metastatic melanoma), renal cell carcinoma, sarcoma, glioma, glioblastoma, multiple myeloma or gastrointestinal cancer, especially colon carcinoma or colorectal adenoma, a tumor of the neck and head, endometrial cancer, Cowden syndrome, Lhermitte-Duclos disease, Bannayan-Zonana syndrome, prostate hyperplasia, a neoplasia, especially of epithelial character, preferably mammary carcinoma or squamous cell carcinoma, chronic lymphocytic leukemia, chronic myelogenous leukemia (e. g., Philadelphia chromosome-positive chronic myelogenous leukemia), acute lymphoblastic leukemia (e. g., Philadelphia chromosome-positive acute lymphoblastic leukemia), non-Hodgkin’s lymphoma, plasma cell myeloma, Hodgkin’s lymphoma, a leukemia, and any combination thereof. 76. A method for the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer, comprising the step of administering the multispecific antibody of any one of items 1 to 53 or the pharmaceutical composition of any one of items 58 to 72 to a patient in need thereof. 77. The method of item 76, wherein the cancer is selected from 119915P877PC 23.12.2024 Numab Therapeutics AG a benign or especially malignant tumor, solid tumors, mesothelioma, brain cancer, kidney cancer, liver cancer, adrenal gland cancer, bladder cancer, breast cancer, stomach cancer (e. g., gastric tumors), esophageal cancer, ovarian cancer, cervical cancer, thymic cancer, choriocarcinoma, oral cancer, salivary cancer, colon cancer, rectum cancer, prostate cancer, pancreatic cancer, lung cancer (e. g., non-small cell lung cancer and small cell lung cancer), vaginal cancer, thyroid cancer, melanoma (e. g., unresectable or metastatic melanoma), renal cell carcinoma, sarcoma, glioma, glioblastoma, multiple myeloma or gastrointestinal cancer, especially colon carcinoma or colorectal adenoma, a tumor of the neck and head, endometrial cancer, Cowden syndrome, Lhermitte-Duclos disease, Bannayan-Zonana syndrome, prostate hyperplasia, a neoplasia, especially of epithelial character, preferably mammary carcinoma or squamous cell carcinoma, chronic lymphocytic leukemia, chronic myelogenous leukemia (e. g., Philadelphia chromosome-positive chronic myelogenous leukemia), acute lymphoblastic leukemia (e. g., Philadelphia chromosome-positive acute lymphoblastic leukemia), non-Hodgkin’s lymphoma, plasma cell myeloma, Hodgkin’s lymphoma, a leukemia, and any combination thereof. 78. An antibody binding domain, having specificity for CD47 (CD47-BD), as defined in any one of items 1, 2, 5, 6, 13 – 16, 21 and 23. 79. The antibody binding domain of item 78, wherein said CD47-BD is in a format selected from an Fv, a dsFv, an scFv and a dsscFv, particularly from an Fv and an scFv, particularly wherein said CD47-BD is in the form of an scFv. 80. An antibody comprising one or two, preferably one, antibody binding domains according to item 78 or 79. 81. An antibody binding domain, having specificity for LILRB2 (LILRB2-BD), as defined in any one of items 40 to 51. 82. The antibody binding domain of item 81, wherein said LILRB2-BD is in a format selected from an Fv, a dsFv, an scFv and a dsscFv, particularly from an Fv and an scFv, particularly wherein said LILRB2-BD is in the form of an scFv. 83. An antibody comprising one, two, three or four, preferably one or two, antibody binding domains according to item 80. 119915P877PC 23.12.2024 Numab Therapeutics AG 84. An antibody according to item 81, which has a sequence selected from: - SEQ ID NO: 207 and SEQ ID NO: 208; - SEQ ID NO: 209 and SEQ ID NO: 210; and - SEQ ID NO: 211 and SEQ ID NO: 212. BRIEF DESCRIPTION OF THE DRAWINGS [0027] Figure 1 shows the binding properties of PRO4710 (IgG4 of scFv PRO3744, 68-18- E11-sc07) to members of LILRBA and LILRB family expressed by CHO cells. [0028] Figure 2 shows the binding properties of PRO3635 (IgG of scFv PRO3064, 68-18- E11-sc02) and the references PRO3483 (NGM), PRO2518 (J19) and PRO3848 (Biond) to members of LILRA family expressed by CHO cells. An anti-V5 tag antibody served as positive control as all constructs contain V5 tag at N-terminus. [0029] Figure 3 shows the binding properties of PRO3635 (IgG of scFv PRO3064, 68-18- E11-sc02) and the references PRO3483 (NGM), PRO2518 (J19) and PRO3848 (Biond) to members of LILRB family expressed by CHO cells. An anti-V5 tag antibody served as positive control as all constructs contain V5 tag at N-terminus. [0030] Figure 4 shows the binding of the scFv PRO3153 (69-01-E02-sc03) applied at concentrations of 150 nM (A) and 50 nM (B) to members of LILRBA and LILRB family expressed by CHO cells. [0031] Figure 5 shows the binding properties of the scFv PRO3697 (69-06-G04-sc05), cross-linked with an anti-framework antibody (x-linked), to members of LILRBA and LILRB family expressed by CHO cells. [0032] Figure 6 shows the dose-dependent increase of the M1 marker CD86 and the dose-dependent decrease of the M2 marker CD163 in the presence of increasing concentrations of PRO3744, PRO3696 and PRO3697. [0033] Figure 7 shows phagocytosis of CFSE-labelled H292 tumor cells by M2-like HMDMs in the presence of increasing concentrations of PRO2777, PRO3599 and PRO3790. Assessment of phagocytosis = % CFSE-positive HMDMs. [0034] Figure 8 shows the results of LPS-driven secretion of IL-1β, TNF-α, and IL-6 in presence of increasing concentrations of multispecific antibodies PRO4672, PRO5052, PRO5053, PRO4687, PRO5055, PRO5056 and anti-LILRB2 antibody PRO4710. [0035] Figure 9 shows the results of LPS-driven secretion of IL-1β, TNF-α, and IL-6 in presence of increasing concentrations of anti-LILRB2 antibody, multispecific antibodies PRO4710, PRO4672, PRO4673, PRO4687, PRO4688, PRO5052, and reference antibodies atezolizumab (Atezo), avelumab, J19.H1 (PRO2518), PRO3599 (Pfizer), and magrolimab (PRO2777). 119915P877PC 23.12.2024 Numab Therapeutics AG [0036] Figure 10 shows the phagocytosis potency of trispecific antibodies PRO4672, PRO4687, PRO5052 and PRO5055 and of the reference compounds PRO2777, PRO3599 and PRO3851 for H292 cancer cells (A) and (C), expressing PDL1 and CD47, and for NALM6 cancer cells (B) and A375 cancer cells (D), expressing only CD47. [0037] Figure 11 shows the phagocytosis potency of trispecific antibodies PRO5052 and PRO5055 and of the reference compounds PRO2777, avelumab, PRO2777 + avelumab combination, PRO5134 (silenced Fc), PRO5054 (no CD47-BD) and PRO5057 (no CD47- BD) for H292 cancer cells (A) and (C), expressing PDL1 and CD47, and for A375 cancer cells (B) and (D), expressing only CD47. [0038] Figure 12 shows the phagocytosis potency of trispecific antibodies PRO5052, PRO5053, PRO5055 and PRO5056 and of the reference compounds PRO2777, PRO2777 + PRO2734 combination, PRO2777 + PRO2734 + avelumab combination, PRO5134 (silenced Fc), PRO5054 (no CD47-BD) and PRO5057 (no CD47-BD) for H292 cancer cells (A) and (C), expressing PDL1 and CD47, and for A375 cancer cells (B) and (D), expressing only CD47. [0039] Figure 13 shows the phagocytosis potency of the trispecific antibodies PRO4672, PRO4687, PRO5055, of the bispecific antibody PRO3790, of the comparative antibodies PRO4682 (CD47-BD in Fab), PRO4711 (CD47-BD from PRO3599), PRO4677 (inverse Morrison) and of the reference compounds PRO2777, PRO3599, for H292 cancer cells (A and C) and red blood cells (RBCs) (B C and D). [0040] Figure 14 shows the phagocytosis potency of the trispecific antibodies PRO4672, PRO4673, PRO4687, PRO4688, of the comparative antibodies PRO4682 and PRO4683 (CD47-BD in Fab), and of the reference antibodies PRO2777, PRO3609 and PRO3613, for H292 cancer cells (A and C) and red blood cells (RBCs) (B and D). [0041] Figure 15 shows the phagocytosis potency of the multispecific antibodies PRO5052, PRO5053, PRO5055 and PRO5056 towards H292 tumor cell and red blood cells (RBCs) in parallel to illustrates the potential therapeutic window. [0042] Figure 16 shows the T-cell proliferation potency of the anti-LILRB2 antibody PRO4710, the trispecific antibodies PRO5052, PRO5053 (no-LILRB2-BD), PRO5055, PRO5056 (no-LILRB2-BD), PRO5054 (no CD47-BD), and the reference antibodies avelumab and magrolimab. [0043] Figure 17 shows the release of TNFα and IL-1β mediated by the anti-LILRB2 antibody PRO4710, the trispecific antibodies PRO4672, PRO4673 (no LILRB2-BD), PRO4687, PRO4688 (no LILRB-BD), PRO5052, PRO5055, PRO5056 (no LILRB2-BD), by the comparative antibodies PRO5054 (no CD47-BD), PRO3613 (medium affinity CD47-BD), and by the reference antibodies magrolimab, atezolizumab (Atezo), PRO3599, PRO3851 and PRO2777. 119915P877PC 23.12.2024 Numab Therapeutics AG [0044] Figure 18 shows the hemagglutination assay result for trispecific antibodies PRO5052, PRO5055, and reference antibodies PRO2777, PRO3599. [0045] Figure 19 shows the hemagglutination assay result for the trispecific antibodies PRO5052, PRO5055, PRO4672, PRO4687, and reference antibodies PRO2777, PRO3851 and PRO3599. [0046] Figure 20 shows the binding of PRO5052, PRO5055 to cancer cell lines HT-29 (A), the binding of PRO5052 and PRO4672 to cancer cell line HT-1080 (B), the binding of PRO4687, PRO5055 and reference antibodies PRO3851 and PRO2777 to cancer cell lines HT-29 (C) and HT-1080 (D). PRO2777 and PRO3851 were applied as reference antibodies. [0047] Figure 21 shows the inhibition of CD47/SIRPα interaction on CHO-CD47 cells (A), NCI-H292 cells (B) and NCI-H292 cells treated with 10 ng/ml INFy (C) for PRO4672, PRO5052 and the reference antibodies PRO2777 and PRO3599. [0048] Figure 22 shows the inhibition of CD47/SIRPα interaction on CHO-CD47 cells (A), NCI-H292 cells (B), NCI-H292 cells treated with 10 ng/ml INFy (C) for PRO4687, PRO5055 and the reference antibodies PRO2777 and PRO3851. [0049] Figure 23 shows the binding properties of PRO5052 (A) and PRO5055 (B) to members of LILRBA and LILRB family expressed by CHO cells. [0050] Figure 24 shows the formats of the multispecific antibodies PRO3790, PRO5052, PRO5053, PRO5055, PRO5056 and the comparative antibodies PRO4677, PRO4682 and PRO4683. DETAILED DESCRIPTION OF THE INVENTION [0051] Known antibody therapeutics targeting CD47 often suffer from dose limiting toxicities, in particular dose limiting toxicities caused by agglutination of red blood cells. Antibody therapeutics targeting PDL1 often lack killing potency even for PDL1-expressing tumor types. Consequently, the development of anti-CD47xPDL1 multispecific antibodies to target cancer cells that use CD47- and PDL1-based escape mechanisms is challenging. There is a need in the medical field to provide anti-CD47xPDL1 antibody-based therapeutics, which provide efficient killing of CD47- and PDL1-expressing cancer cells, while exhibiting low or no dose limiting toxicities. [0052] The present invention provides multispecific antibodies comprising one CD47-BD and one PDL1-BD, and an IgG-like region comprising an immunoglobulin Fc region, a first Fab arm and a second Fab arm, wherein the CD47 BD is an scFv fragment replacing the first Fab arm, and the PDL1 BD is located in the second Fab arm or is an scFv fragment replacing said second Fab arm. These multispecific antibodies do not cause phagocytosis of red blood cells or trigger any visible red blood cell agglutination even at high nanomolar 119915P877PC 23.12.2024 Numab Therapeutics AG concentrations. At the same time, they exhibit a high potency in killing cancer cells that express CD47 and PDL1. [0053] It is believed by the present inventors that the observed selectivity, and in particular the absence of red blood cell agglutination and red blood cell lysis at nanomolar concentrations, is due to the presence of moderately affine CD47-BD, in combination with a well-defined IgG-like antibody architecture, wherein the CD47-BD is present in the form of an scFv. [0054] Alternative antibody architectures, e. g. antibody architectures wherein said CD47- BD is not in scFv format and/or the PDL1-BD and the CD47-BD are not located next to each other in the Fab arms, do not exhibit the desired combined properties, i. e. potent cancer cell lysis and at the same time low or no agglutination/lysis of red blood cells. For example, antibody formats wherein the CD47-BD is in Fab format exhibit significant red blood cell agglutination. IgG like anti-PDL1 x CD47 antibody formats, where the PDL1-BD and CD47- BD is placed at the C-terminus of the Fc region, still exhibit low to no red blood cell agglutination, but lose the ability to kill cancer cells. Likewise, Fc silenced IgG-like formats or Fc less antibody formats, e. g. Numab’s MATCH formats, do not provide the desired cancer cell killing potency. The presence of a Fc region, which has at least some Fc receptor- binding functionality, is apparently important for efficient cancer cell killing. [0055] It has further been found that the exemplified multispecific antibodies of the present invention, e. g. PRO3790, PRO4672, PRO4673, PRO4687, PRO4688, PRO5052, PRO5053, PRO5055, PRO5056, exhibit lower red blood cell agglutination and/or red blood cell lysis than the prior art anti-CD47xPDL1 antibodies PF-07257876 (Pfizer Inc.) and IBI-322 (Innovent Biologics Inc), while exhibiting similar or even better in-vitro cancer cell killing potency. It is thus expected that the multispecific antibodies of the present invention will provide a large therapeutic window and thus provide distinct therapeutic advantages over conventional therapies. [0056] In addition, the multispecific antibodies of the present invention exhibit very advantageous biophysical properties, in particular high storage stability. For example, the loss of monomeric content for PRO4672, PRO5052, PRO4687 and PRO5055, which are representative multispecific antibodies of the present invention, is less than 1 % when stored for two months at 4°C at a concentration of 10 mg/ml in a 20 mM Histidine buffer (pH 6.0) with 125 mM NaCl and 0.02 % Poloxamer 188. This high stability is particularly surprising in view of the fact that the multispecific antibodies of the invention comprise up to four scFv antibody fragments. [0057] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which this invention pertains. 119915P877PC 23.12.2024 Numab Therapeutics AG [0058] The terms “comprising” and “including” are used herein in their open-ended and non-limiting sense unless otherwise noted. With respect to such latter embodiments, the term "comprising" thus includes the narrower term “consisting of”. [0059] The terms “a” and “an” and “the” and similar references in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. For example, the term “a cell” includes a plurality of cells, including mixtures thereof. Where the plural form is used for compounds, salts, and the like, this is taken to mean also a single compound, salt, or the like. [0060] The term “antibody” and the like, as used herein, includes whole antibodies or single chains thereof; and any antigen-binding fragment (i. e., “antigen-binding portion”) or single chains thereof; and molecules comprising antibody CDRs, VH regions or VL regions (including without limitation multispecific antibodies). A naturally occurring “whole antibody” is a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter- connected by disulfide bonds. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is comprised of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), flanked by regions that are more conserved, termed framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e. g., effector cells) and the first component (C1q) of the classical complement system. [0061] The term “immunoglobulin Fc region” or “Fc region”, as used herein, is used to define a C-terminal region of an immunoglobulin heavy chain, i. e. the CH2 and CH3 domains of the heavy chain constant regions. The Fc region provides linkage to humoral and cellular components of the immune system, mostly through interaction with C1q and Fc receptors (FcR). The term “Fc region” includes Fc regions of an IgG subclass, particularly an immunoglobulin Fc region of human lgG1, lgG2 (lgG2A, IgG2B), lgG3 or lgG4. Further included are Fc regions that are engineered, i. e. Fc regions that have one or more amino acid modifications, to exhibit certain desired properties. Such amino acid modifications are for example modifications to enhance heterodimerization of heavy chains, such as knob-into- 119915P877PC 23.12.2024 Numab Therapeutics AG hole (KiH) technology (see for example Ridgway et al., Protein Eng.9:617-21 (1996) and Spiess et al., J Biol Chem.288(37):26583-93 (2013)). Other amino acid modifications are modifications that alter, i. e. increase or decrease Fc receptor-binding, when compared to the respective unmodified immunoglobulin Fc region thereof (see for example the modifications disclosed in WO 2004/42072 (Presta) and in Wang et al., Protein Cell.9(1):63- 732018 (2018)) and/or increase serum half-life (see for example the modifications disclosed in Liu et al., Antibodies (Basel).9(4):64 (2020)). [0062] The term “Fc receptor” or “FcR” describes a receptor that binds to the Fc region of an antibody. Particularly, the Fc receptors are native sequence human FcRs, which binds an IgG antibody. These include for example the Fc gamma receptors (FcγRs). Five activating FcγRs are known, i. e. the high affinity receptors FcγRI, and the lower affinity receptors FcγRIIA, FcγRIIC, FcγRIIIA and FcγRIIIB. Besides, one inhibiting receptor is known, i. e. FcγRIIB, which have similar amino acid sequences to FcγRIIA that differ primarily in the cytoplasmic domains thereof. Activating receptors contain an immunoreceptor tyrosine- based activation motif (ITAM) in its cytoplasmic domain. The inhibiting receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain, (see M. Daeron, Annu. Rev. Immunol.5:203-234 (1997). FcRs are reviewed for example in Ravetch and Kinet, Annu. Rev. Immunol.9: 457-92 (1991); Capet et al, Immunomethods 4: 25-34 (1994); and de Haas et al, J. Lab. Clin. Med.126: 330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term “FcR” herein. The term “Fc receptor” or “FcR” also includes the neonatal receptor FcRn, which plays a central role in the cellular trafficking and serum half-life of IgGs. High-affinity binding to FcRn in vivo is reported to prolong serum half-life of human IgGs (see for example Dall’Acqua et al., J Biol Chem.281:23514–23524(2006)). [0063] The term “silenced Fc”, “Fc silenced” or “silenced Fc region", as used herein, refers to an Fc region, which has been engineered to exhibit insignificant or negligible Fc receptor- binding functionality, in particular insignificant or negligible binding affinity to FcγRs, when compared to its unmodified version. Antibodies having such silenced Fc regions do not or do not significantly trigger cellular immune responses through Fc receptor-binding. [0064] The term “active Fc” or “active Fc region", as used herein, refers to an Fc region, which has significant Fc receptor-binding functionality, in particular significant binding affinity to FcγRs. This Fc receptor-binding functionality may be present naturally and/or may be introduced or enhanced by engineering. Antibodies having such active Fc regions trigger cellular immune responses through Fc receptor-binding. [0065] In particular embodiments, the immunoglobulin Fc region comprised in the multispecific antibodies of the invention is selected from an Fc region of an IgG subclass, 119915P877PC 23.12.2024 Numab Therapeutics AG particularly an immunoglobulin Fc region of human lgG1, lgG2 (lgG2A, IgG2B), lgG3 or lgG4, particularly of human IgG1 or IgG2, particularly of human IgG1. [0066] In some embodiments, the immunoglobulin Fc region comprised in the multispecific antibodies of the invention is an active Fc region. [0067] In further embodiments, the immunoglobulin Fc region comprised in the multispecific antibodies of the invention has one or more amino acid modifications selected from the group consisting of: a) modifications to enhance heterodimerization of heavy chains, such as the knob-into-hole (KiH) technology; b) modifications that increase or decrease, in particular increase, Fc receptor-binding, when compared to the respective unmodified immunoglobulin Fc region thereof; c) modifications that increase serum half-life, when compared to the respective unmodified immunoglobulin Fc region thereof. [0068] In particular embodiments, the immunoglobulin Fc region comprised in the multispecific antibodies of the invention has one or more amino acid modifications to enhance heterodimerization of the heavy chains, such as the knob-into-hole (KiH) technology. [0069] The terms “binding domain”, “antigen-binding fragment thereof”, “antigen-binding portion” of an antibody, and the like, as used herein, refer to one or more fragments of an intact antibody that retain the ability to specifically bind to a given antigen (e. g., CD47, PDL1, LILRB2). Antigen-binding functions of an antibody can be performed by fragments of an intact antibody. Specifically, in case of the multispecific antibodies of the present invention, the terms “binding domain”, “antigen-binding fragment”, “antigen-binding fragment thereof”, “antigen-binding portion”, and the like, as used herein, refer to a Fab fragment, i. e. a monovalent fragment consisting of the VL, VH, CL and CH1 domains; a single chain Fab, i. e. a Fab fragment, where the heavy chain and the light chain are connected via a linker; an Fv fragment consisting of the VL and VH domains of a single arm of an antibody; a disulfide stabilized Fv fragment (dsFv); and a single chain Fv fragment (scFv). Preferably, the binding domains of the antibodies of the present invention are independently of each other selected from a Fab fragment, an Fv fragment, a dsFv fragment and a single-chain Fv fragment (scFv). In particular embodiments, the binding domains of the antibodies of the present invention are independently of each other selected from a Fab fragment and a single-chain Fv fragment (scFv). In other particular embodiments, the VL and VH domains of the scFv fragment are stabilized by an interdomain disulfide bond (dsscFv), in particular said VH domain comprises a single cysteine residue in position 51 (AHo numbering) and said VL domain comprises a single cysteine residue in position 141 (AHo numbering). 119915P877PC 23.12.2024 Numab Therapeutics AG [0070] The term “fragment-based” or “fragment-based antibody”, as used herein, refers to an antibody that comprises at least one antigen-binding fragment, as defined above, but no immunoglobulin Fc region. Due to the lack of the Fc region, these antibodies typically comprise and hSA-binding domain for half-life extension. [0071] The term “Complementarity Determining Regions” (“CDRs”) refers to amino acid sequences with boundaries determined using any of a number of well-known schemes, including those described by Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (“Kabat” numbering scheme); Al-Lazikani et al., (1997) JMB 273, 927-948 (“Chothia” numbering scheme); ImMunoGenTics (IMGT) numbering (Lefranc, M.-P., The Immunologist, 7, 132-136 (1999); Lefranc, M.-P. et al., Dev. Comp. Immunol., 27, 55-77 (2003)) (“IMGT” numbering scheme); and the numbering scheme described in Honegger & Plückthun, J. Mol. Biol.309 (2001) 657-670 (“AHo” numbering). For example, for classic formats, under Kabat, the CDR amino acid residues in the heavy chain variable region (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable region (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). Under Chothia the CDR amino acids in the VH are numbered 26-32 (HCDR1), 52-56 (HCDR2), and 95-102 (HCDR3); and the amino acid residues in VL are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). By combining the CDR definitions of both Kabat and Chothia, the CDRs consist of amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in human VH and amino acid residues 24-34 (LCDR1), 50- 56 (LCDR2), and 89-97 (LCDR3) in human VL. Under IMGT the CDR amino acid residues in the VH are numbered approximately 26-35 (HCDR1), 51-57 (HCDR2) and 93-102 (HCDR3), and the CDR amino acid residues in the VL are numbered approximately 27-32 (LCDR1), 50-52 (LCDR2), and 89-97 (LCDR3) (numbering according to “Kabat”). Under IMGT, the CDRs of an antibody can be determined using the program IMGT/DomainGap Align. [0072] In the context of the present invention, the numbering system suggested by Honegger & Plückthun (“AHo”) is used (Honegger & Plückthun, J. Mol. Biol.309 (2001) 657- 670), unless specifically mentioned otherwise. In particular, the following residues are defined as CDRs according to AHo numbering scheme: LCDR1 (also referred to as CDR- L1): L24-L42; LCDR2 (also referred to as CDR-L2): L58-L72; LCDR3 (also referred to as CDR-L3): L107-L138; HCDR1 (also referred to as CDR-H1): H27-H42; HCDR2 (also referred to as CDR-H2): H57-H76; HCDR3 (also referred to as CDR-H3): H108-H138. For the sake of clarity, the numbering system according to Honegger & Plückthun takes the length diversity into account that is found in naturally occurring antibodies, both in the different VH and VL subfamilies and, in particular, in the CDRs, and provides for gaps in the 119915P877PC 23.12.2024 Numab Therapeutics AG sequences. Thus, in a given antibody variable region usually not all positions 1 to 149 will be occupied by an amino acid residue. [0073] The term “binding specificity” as used herein refers to the ability of an individual antibody to react with one antigenic determinant and not with a different antigenic determinant. As used herein, the term “specifically binds to” or is “specific for” refers to measurable and reproducible interactions such as binding between a target and an antibody, which is determinative of the presence of the target in the presence of a heterogeneous population of molecules including biological molecules. For example, an antibody that specifically binds to a target (which can be an epitope) is an antibody that binds this target with greater affinity, avidity, more readily, and/or with greater duration than it binds to other targets. In its most general form (and when no defined reference is mentioned), “specific binding” is referring to the ability of the antibody to discriminate between the target of interest and an unrelated molecule, as determined, for example, in accordance with specificity assay methods known in the art. Such methods comprise, but are not limited to Western blots, ELISA, RIA, ECL, IRMA, SPR (Surface plasmon resonance) tests and peptide scans. For example, a standard ELISA assay can be carried out. The scoring may be carried out by standard color development (e. g. secondary antibody with horseradish peroxide and tetramethyl benzidine with hydrogen peroxide). The reaction in certain wells is scored by the optical density, for example, at 450 nm. Typical background (= negative reaction) may be about 0.1 OD; typical positive reaction may be about 1 OD. This means the ratio between a positive and a negative score can be 10-fold or higher. In a further example, an SPR assay can be carried out, wherein at least 10-fold, particularly at least 100-fold difference between a background and signal indicates on specific binding. Typically, determination of binding specificity is performed by using not a single reference molecule, but a set of about three to five unrelated molecules, such as milk powder, transferrin or the like. [0074] Suitably, the antibodies of the invention are isolated antibodies. The term “isolated antibody”, as used herein, refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e. g., an isolated antibody that specifically binds CD47 and PDL1 is substantially free of antibodies that specifically bind antigens other than CD47 and PDL1. Moreover, an isolated antibody may be substantially free of other cellular material and/or chemicals. [0075] Suitably, the antibodies of the invention are monoclonal antibodies. The term “monoclonal antibody” as used herein refers to antibodies that have substantially identical amino acid sequences or are derived from the same genetic source. A monoclonal antibody displays a binding specificity and affinity for a particular epitope, or binding specificities and affinities for specific epitopes. 119915P877PC 23.12.2024 Numab Therapeutics AG [0076] Antibodies of the invention include, but are not limited to, chimeric, human and humanized antibodies. [0077] The term “chimeric antibody” (or antigen-binding fragment thereof), as used herein, refers to an antibody molecule (or antigen-binding fragment thereof) in which (a) the constant region, or a portion thereof, is altered, replaced or exchanged so that the antigen- binding site (variable region) is linked to a constant region of a different or altered class, effector function and/or species; or (b) the variable region, or a portion thereof, is altered, replaced or exchanged with a variable region having a different or altered antigen specificity. For example, a mouse antibody can be modified by replacing its constant region with the constant region from a human immunoglobulin. Due to the replacement with a human constant region, the chimeric antibody can retain its specificity in recognizing the antigen while having reduced antigenicity in human as compared to the original mouse antibody. [0078] The term “human antibody” (or antigen-binding fragment thereof), as used herein, is intended to include antibodies (and antigen-binding fragments thereof) having variable regions in which both the framework and CDR regions are derived from sequences of human origin. Furthermore, if the antibody contains a constant region, the constant region also is derived from such human sequences, e. g., human germline sequences, or mutated versions of human germline sequences. The human antibodies and antigen-binding fragments thereof of the invention may include amino acid residues not encoded by human sequences (e. g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phage-display libraries (Hoogenboom and Winter, J. Mol. Biol, 227:381 (1992); Marks et al, J. Mol. Biol, 222:581 (1991)). Also available for the preparation of human monoclonal antibodies are methods described in Cole et al, Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p.77 (1985); Boemer et al, J. Immunol, 147(l):86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol, 5: 368-74 (2001). Human antibodies can be prepared by administering the antigen to a transgenic animal that has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled, e. g., immunized xenomice (see, e. g., U.S. Pat. Nos.6,075,181 and 6,150,584 regarding XENOMOUSE™ technology). See also, for example, Li et al, Proc. Natl. Acad. Sci. USA, 103:3557- 3562 (2006) regarding human antibodies generated via a human B-cell hybridoma technology. [0079] The term “humanized” antibody (or antigen-binding fragment thereof), as used herein, refers to an antibody (or antigen-binding fragment thereof) that retains the reactivity of a non-human antibody while being less immunogenic in humans. This can be achieved, 119915P877PC 23.12.2024 Numab Therapeutics AG for instance, by retaining the non-human CDR regions and replacing the remaining parts of the antibody with their human counterparts (i. e., the constant region as well as the framework portions of the variable region). Additional framework region modifications may be made within the human framework sequences as well as within the CDR sequences derived from the germline of another mammalian species. In particular, it is known that CDR1 of VH and CDR2 of VL are less important for binding specificity, as evidenced by the fact that in many antibodies obtained by immunization, those CDRs are identical to the CDR encoded by the corresponding germline gene segment. Thus, in one embodiment of the invention, the antibody or antigen-binding fragment thereof comprises in CDR1 of VH and/or in CDR2 of VL one or two amino acid residues that differ from the amino acid residues of the parental antibody. The humanized antibodies of the invention may include amino acid residues not encoded by human sequences (e. g., mutations introduced by random or site- specific mutagenesis in vitro or by somatic mutation in vivo, or a conservative substitution to promote stability or manufacturing). See, e. g., Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855, 1984; Morrison and Oi, Adv. Immunol., 44:65-92, 1988; Verhoeyen et al., Science, 239: 1534-1536, 1988; Padlan, Molec. Immun., 28:489-498, 1991; and Padlan, Molec. Immun., 31: 169-217, 1994. Other examples of human engineering technology include but are not limited to the Xoma technology disclosed in U.S. Pat. No.5,766,886. [0080] The term “recombinant humanized antibody” as used herein, includes all human antibodies that are prepared, expressed, created or isolated by recombinant means, such as antibodies isolated from a host cell transformed to express the humanized antibody, e. g., from a transfectoma, and antibodies prepared, expressed, created or isolated by any other means that involve splicing of all or a portion of a human immunoglobulin gene, sequences to other DNA sequences. [0081] Preferably, the multispecific antibodies of the invention are humanized. More preferably, the multispecific antibodies of the invention are humanized and comprises rabbit derived CDRs. [0082] The term “multispecific antibody” as used herein, refers to an antibody that binds to two or more different epitopes on at least two or more different targets (e. g., CD47, PDL1, hSA and LILRB2). Preferably, the multispecific antibodies of the invention are bispecific or trispecific. The term “bispecific antibody” as used herein, refers to an antibody that binds to at least two different epitopes on two different targets (e. g., CD47 and PDL1). The term “trispecific antibody” as used herein, refers to an antibody that binds to at least three different epitopes on three different targets (e. g., CD47, PDL1, hSA and LILRB2). [0083] The term “epitope” means a protein determinant capable of specific binding to an antibody. Epitopes usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three-dimensional structural 119915P877PC 23.12.2024 Numab Therapeutics AG characteristics, as well as specific charge characteristics. “Conformational” and “linear” epitopes are distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents. [0084] The term “conformational epitope” as used herein refers to amino acid residues of an antigen that come together on the surface when the polypeptide chain folds to form the native protein. [0085] The term “linear epitope” refers to an epitope, wherein all points of interaction between the protein and the interacting molecule (such as an antibody) occurring linearly along the primary amino acid sequence of the protein (continuous). [0086] The term “recognize” as used herein refers to an antibody antigen-binding fragment thereof that finds and interacts (e. g., binds) with its conformational epitope. [0087] The term “avidity” as used herein refers to an informative measure of the overall stability or strength of the antibody-antigen complex. It is controlled by three major factors: antibody epitope affinity; the valency of both the antigen and antibody; and the structural arrangement of the interacting parts. Ultimately these factors define the specificity of the antibody, that is, the likelihood that the particular antibody is binding to a precise antigen epitope. [0088] Suitably, the multispecific antibodies of the invention comprise one CD47 biding domain. [0089] The term “CD47” refers in particular to human CD47 with UniProt ID number Q08722. Suitably, the CD47-BD of the present invention targets CD47, in particular human CD47. Particularly, the multispecific antibodies of the invention comprises one CD47-BD that target human and cynomolgus (Macaca fascicularis) CD47. [0090] Suitably, the CD47-BD, when in scFv format, binds to human CD47 with a monovalent dissociation constant (KD) of 1 to 100 nM, particularly with a KD of 2 to 50 nM, particularly of 5 to 25 nM, as measured by surface plasmon resonance (SPR). [0091] As used herein, the term “affinity” refers to the strength of interaction between the antibody and the antigen at single antigenic sites. Within each antigenic site, the variable region of the antibody “arm” interacts through weak non-covalent forces with antigen at numerous sites; the more interactions, the stronger the affinity. [0092] “Binding affinity” generally refers to the strength of the total sum of non-covalent interactions between a single binding site of a molecule (e. g., of an antibody) and its binding partner (e. g., an antigen or, more specifically, an epitope on an antigen). Unless indicated otherwise, as used herein, “binding affinity”, “bind to”, “binds to” or “binding to” refers to intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e. g., an antibody fragment and an antigen). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (KD). Affinity can be measured by 119915P877PC 23.12.2024 Numab Therapeutics AG common methods known in the art, including those described herein. Low-affinity antibodies generally bind antigens slowly and tend to dissociate readily, whereas high-affinity antibodies generally bind antigens faster and tend to remain bound longer. A variety of methods of measuring binding affinity are known in the art, any of which can be used for purposes of the present invention. Specific illustrative and exemplary embodiments for measuring binding affinity, i. e. binding strength are described in the following. [0093] The term “Kassoc”, “Ka” or “Kon”, as used herein, are intended to refer to the association rate of a particular antibody-antigen interaction, whereas the term “Kdis”, “Kd” or “Koff”, as used herein, is intended to refer to the dissociation rate of a particular antibody- antigen interaction. In one embodiment, the term “KD”, as used herein, is intended to refer to the dissociation constant, which is obtained from the ratio of Kd to Ka (i. e. Kd/Ka) and is expressed as a molar concentration (M). The “KD” or “KD value” or "KD" or "KD value" according to this invention is in one embodiment measured by using surface plasmon resonance assays. [0094] Affinity to recombinant human CD47 and recombinant cynomolgus CD47 was determined by surface plasmon resonance (SPR) measurements, as described in Example 2.6 (scFvs) and Examples 4.2 and 5.3 (multispecific molecules). Affinity to recombinant human PDL1 and recombinant cynomolgus PDL1 was determined by surface plasmon resonance (SPR) measurements, as described in Example 1.4 (scFvs) and Examples 4.2 and 5.3 (multispecific molecules). Likewise, affinity to recombinant human LILRB2 and recombinant cynomolgus LILRB2 was determined by surface plasmon resonance (SPR) measurements, as described in Example 3.6 (scFvs) and Example 5.3 (multispecific molecules). [0095] Suitably, the multispecific antibody of the invention acts as a blocker of the CD47 interaction with SIRPα. In other words, the CD47-BD of the multispecific antibody of the invention is an inhibitor of CD47 signaling. The term “blocker” or “inhibitor” or “antagonist”, as used herein, refers to an antibody or binding domain thereof that inhibits or reduces a biological activity of the antigen it binds to. The CD47-BD of the multispecific antibody of the invention bind to CD47, thereby blocking the binding of CD47 to SIRPα, which leads to reduced CD47 function. [0096] Suitably, the CD47-BD of the multispecific antibody of the present invention is further characterized by one or more of the following parameters: a. when in scFv format, has a melting temperature (Tm), determined by differential scanning fluorimetry, in the range of 50°C to 70°C, particularly in the range of 55°C to 65°C, in particular wherein said antibody is formulated in 50 mM phosphate-citrate buffer (NaCiP) at pH 6.4, 150 mM NaCl; 119915P877PC 23.12.2024 Numab Therapeutics AG b. when in scFv format, has a loss in monomer content, after storage for four weeks at ‑80°C, of less than 3 %, less than 2 %, less than 1 %, particularly less than 0.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer (NaCiP) with 150 mM NaCl at pH 6.4; c. when in scFv format, has a loss in monomer content, after storage for four weeks at 4°C, of less than 3 %, less than 2 %, less than 1 %, particularly less than 0.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer (NaCiP) with 150 mM NaCl at pH 6.4; and/or d. when in scFv format, has a loss in monomer content, after storage for four weeks at 40°C, of less than 3 %, less than 2 %, less than 1 %, particularly less than 0.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer (NaCiP) with 150 mM NaCl at pH 6.4. [0097] Differential scanning fluorimetry (DSF) measurements are performed as described earlier (Egan, et al., MAbs, 9(1) (2017), 68-84; Niesen, et al., Nature Protocols, 2(9) (2007) 2212-2221). In this DSF measurement procedure, the midpoint of transition for the thermal unfolding of the scFv constructs is determined by using the fluorescence dye SYPRO Orange (see Wong & Raleigh, Protein Science 25 (2016) 1834-1840). Typically, samples in phosphate-citrate buffer at pH 6.4 are prepared at a final protein concentration of 50 µg/ml and containing a final concentration of 5x SYPRO Orange in a total volume of 100 µl. Twenty-five microliters of prepared samples are added in triplicate to white-walled AB gene PCR plates. The assay is performed in a qPCR machine used as a thermal cycler, and the fluorescence emission is detected using the software’s custom dye calibration routine. The PCR plate containing the test samples is subjected to a temperature ramp from 25°C to 96°C in increments of 1°C with 30 s pauses after each temperature increment. The total assay time is about 2 h. The Tm is calculated by the software GraphPad Prism using a mathematical second derivative method to calculate the inflection point of the curve. The reported Tm is typically an average of three measurements. [0098] The loss in monomer content is as determined by area under the curve calculation of SE-HPLC chromatograms. SE-HPLC is a separation technique based on a solid stationary phase and a liquid mobile phase as outlined by the US Pharmacopeia (USP), chapter 621. This method separates molecules based on their size and shape utilizing a hydrophobic stationary phase and aqueous mobile phase. The separation of molecules is occurring between the void volume (V0) and the total permeation volume (VT) of a specific 119915P877PC 23.12.2024 Numab Therapeutics AG column. Measurements by SE-HPLC are performed on a Chromaster HPLC system (Hitachi High-Technologies Corporation) equipped with automated sample injection and a UV detector set to the detection wavelength of 280 nm. The equipment is controlled by the software EZChrom Elite (Agilent Technologies, Version 3.3.2 SP2) which also supports analysis of resulting chromatograms. Protein samples are cleared by centrifugation and kept at a temperature of 4-6°C in the autosampler prior to injection. For the analysis of scFv samples the column Shodex KW403-4F (Showa Denko Inc., #F6989202) is employed with a standardized buffered saline mobile phase (50 mM sodium-phosphate pH 6.5, 300 mM sodium chloride) at the recommended flow rate of 0.35 ml/min. The target sample load per injection was 5 µg. Samples are detected by an UV detector at a wavelength of 280 nm and the data recorded by a suitable software suite. The resulting chromatograms are analyzed in the range of V0 to VT thereby excluding matrix associated peaks with >10 min elution time. [0099] Suitably, the PDL1-BDs of the multispecific antibodies of the invention are binding domains provided in the present disclosure. The PDL1-BDs of the multispecific antibodies of the invention include, but are not limited to, the humanized PDL1-BDs whose sequences are listed in Table 1. [0100] Suitably, the multispecific antibodies of the invention comprise one PDL1-BD. [0101] The term “PDL1” or “PD-L1” refers in particular to human PDL1 with UniProt ID number Q9NZQ7. Suitably, the PDL1-BD of the multispecific antibodies of the invention targets, in particular human PDL1. Particularly, the multispecific antibodies of the invention comprise one PDL1-BD that targets human and cynomolgus (Macaca fascicularis) PDL1. [0102] Suitably, the PDL1-BD used in the present invention is characterized by one or more of the following parameters: a. when in scFv format, binds to human PDL1 with a monovalent dissociation constant (KD) of 1 to 100 pM, particularly with a KD of 1 to 50 pM, particularly of 1 to 40 pM, as measured by surface plasmon resonance (SPR); b. when in scFv format, is cross-reactive with Macaca fascicularis (cynomolgus) PDL1, in particular binds to cynomolgus PDL1 with a monovalent KD of 1 to 100 pM, particularly with a KD of 1 to 50 pM, particularly of 1 to 40 pM, as measured by SPR; c. when in scFv format, has the ability to neutralize PDL1/PD-1 interaction, as measured in an NFAT reporter gene assay, with an EC50 value of 0.1 to 10 nM, particularly of 0.1 to 5 pM, particularly of 0.1 to 3 nM; d. is non-cross-reactive to Mus musculus PDL1, as measured by SPR; and/or e. does not bind to human PDL2, as measured by SPR. [0103] Suitably, the PDL1-BD of the multispecific antibodies of the present invention is further characterized by one or more of the following parameters: a. when in scFv format, has a melting temperature (Tm), determined by differential scanning fluorimetry, in the range of 60°C to 85°C, particularly in the range of 65°C to 119915P877PC 23.12.2024 Numab Therapeutics AG 85°C, particularly in the range of 70°C to 85°C, in particular wherein said antibody is formulated in 50 mM phosphate-citrate buffer at pH 6.4, 150 mM NaCl; b. when in scFv format, has a loss in monomer content, after five consecutive freeze-thaw cycles, of less than 5 %, preferably less than 3 %, more preferably less than 1 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein the antibody of the invention is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4; and/or c. when in scFv format, has a loss in monomer content, after storage for four weeks at 4°C, of less than 15 %, e. g. less than 12 %, less than 10 %, less than 7 %, less than 5 %, less than 4 %, less than 3 %, less than 2 %, preferably less than 1 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4. [0104] Suitably, the PDL1-BDs of the multispecific antibodies of the invention are binding domains provided in the present disclosure. The PDL1-BDs of the multispecific antibodies of the invention include, but are not limited to, the humanized PDL1-BDs whose sequences are listed in Table 1. [0105] The term “multivalent antibody” refers to a single binding molecule with more than one valency, where “valency” is described as the number of antigen-binding moieties that binds to epitopes on target molecules. As such, the single binding molecule can bind to more than one binding site on a target molecule and/or to more than one target molecule due to the presence of more than one copy of the corresponding antigen-binding moieties. Examples of multivalent antibodies include, but are not limited to bivalent antibodies, trivalent antibodies, tetravalent antibodies, pentavalent antibodies, hexavalent antibodies, and the like. [0106] The term “monovalent antibody”, as used herein, refers to an antibody that binds to a single target molecule, and more specifically to a single epitope on a target molecule. Also, the term “binding domain” or “monovalent binding domain”, as used herein, refers to a binding domain that binds to a single epitope on a target molecule. [0107] The multispecific antibodies of the invention comprise one CD47-BD and one PDL1-BD, i. e. the multispecific antibodies of the invention are monovalent for both CD47 and PDL1 specificity. [0108] In particular embodiments, the KD of the PDL1-BD for binding to human PDL1 is at least 500 fold lower, in particular at least 1000 fold lower, in particular at least 1500 fold lower, in particular at least 2000 fold lower, than the KD of the CD47-BD for binding to human CD47. 119915P877PC 23.12.2024 Numab Therapeutics AG [0109] The multispecific antibodies of the invention do not trigger any visible red blood cell agglutination at a concentration of 5 nM, particularly of 20 nM, particularly of 50 nM, particularly of 100 nM, particularly of 200 nM, particularly of 500 nM, as determined in a standard hemagglutination assay as described in Example 4.2 [0110] The expression “hemagglutination” as used herein, refers to a specific form of agglutination that involves red blood cells (RBCs). More specifically, the expression hemagglutination refers to agglutination of RBCs that is caused by antibodies that bind to surface antigens on the RBCs. Hemagglutination is thus a sensitive method to detect RBC- binding antibodies. The assay procedure to detect such hemagglutination is described in Example 4.2. [0111] Likewise, the multispecific antibodies of the invention do not cause the lysis of red blood cells at a concentration of 5 nM, particularly of 20 nM, particularly of 50 nM, as determined in a standard phagocytosis assay, as described in Example 5.4. [0112] Antibody-dependent cell-mediated cytotoxicity (ADCC) of red blood cells, herein also referred to as “lysis of red blood cell” or “red blood cell phagocytosis”, is typically determine analogous to the ADCC of other cells where antibodies can bind to. More specifically, the red blood cell lysis was determined analogous to the lysis of the CD47 expressing or CD47/PDL1-coexpressing cell, as described in Example 5.4. [0113] In a particular aspect, the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequences of SEQ ID NO: 7; or ii. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequences of SEQ ID NO: 7; b) one binding domain, which specifically binds to PDL1 (PDL1-BD); wherein said PDL1-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 21, 119915P877PC 23.12.2024 Numab Therapeutics AG the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequences of SEQ ID NO: 27; or ii. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 21, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequences of SEQ ID NO: 27; or iii. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequences of SEQ ID NO: 27; or iv. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequences of SEQ ID NO: 27; or v. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 78, and the LCDR3 sequences of SEQ ID NO: 80; or vi. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 79, and the LCDR3 sequences of SEQ ID NO: 80; c) an IgG-like region comprising an immunoglobulin Fc region, a first Fab arm and a second Fab arm, wherein: - the CD47 BD is an scFv fragment replacing the first Fab arm, and 119915P877PC 23.12.2024 Numab Therapeutics AG - the PDL1 BD is located in the second Fab arm or is an scFv fragment replacing said second Fab arm. [0114] In a more particular aspect, the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequences of SEQ ID NO: 7; or ii. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequences of SEQ ID NO: 7; b) one binding domain, which specifically binds to PDL1 (PDL1-BD); wherein said PDL1-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 78, and the LCDR3 sequences of SEQ ID NO: 80; or ii. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 79, and the LCDR3 sequences of SEQ ID NO: 80; c) an IgG-like region comprising an immunoglobulin Fc region, a first Fab arm and a second Fab arm, wherein: - the CD47 BD is an scFv fragment replacing the first Fab arm, and - the PDL1 BD is located in the second Fab arm or is an scFv fragment replacing said second Fab arm. 119915P877PC 23.12.2024 Numab Therapeutics AG [0115] Other variable regions used in the invention include amino acid sequences that have been mutated, yet have at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identity in the CDR regions with the CDR regions depicted in the sequences described in Table 1. Other variable regions used the invention include mutant amino acid sequences wherein no more than 1, 2, 3, 4 or 5, particularly 1 or 2, amino acids have been mutated in the CDR regions when compared with the CDR regions depicted in the sequence described in Table 1. [0116] Suitably, the VH domains of the binding domains of the multispecific antibodies of the invention, e. g. the CD47-BD, PDL1-BD, hSA-BD or LILRB2-BD, belong to a VH3 or VH4 family. In one embodiment, a binding domain used in the invention comprises a VH domain belonging to the VH3 family. [0117] In the context of the present invention, the term “belonging to VHx family” or “selected from VHx family” means that the respective heavy chain framework sequence or sequences referred to, e. g. FR1, FR2, FR3, FR4, FR1 to FR3, FR1 to FR4, show the highest degree of homology to said VHx family. Likewise, the term “belonging to Vx family” or “selected from Vx family” means that the respective light chain framework sequence or sequences referred to, e. g. FR1, FR2, FR3, FR4, FR1 to FR3, FR1 to FR4, show the highest degree of homology to said Vx family. Examples of VH and VL families are given in Knappik et al., J. Mol. Biol.296 (2000) 57-86, or in WO 2019/057787. [0118] A specific example of a VH domain belonging to VH3 family is represented by SEQ ID NOs: 151 and 152, and a specific example of a VH domain belonging to VH4 family is represented by SEQ ID NO: 155. Particularly, a VH belonging to VH3 family, as used herein, wherein each FR1 to FR4, independently of each other, have at least 85 %, particularly at least 90 %, more particularly at least 95 % sequence identity to the corresponding FR1 to FR4 of SEQ ID NO: 151 or 152. In particular, heavy chain framework regions FR1 to FR4 are selected from SEQ ID NOs: 151 and 152 (Table 2, regions marked in non-bold). Alternative examples of VH3 and VH4 sequences, and examples of other VHx sequences, may be found in Knappik et al., J. Mol. Biol.296 (2000) 57-86 or in WO 2019/057787. [0119] Suitably, the VL domains of the binding domains used in the invention comprise: Vκ frameworks FR1 to FR4, particularly Vκ1 or Vκ3 frameworks FR1 to FR4, particularly Vκ1 frameworks FR1 to FR4. In the case where said binding domains are not in a Fab format, for example where said binding domains are in an scFv-format, said binding domains comprise: Vκ frameworks FR1 to FR3, particularly Vκ1 or Vκ3 frameworks FR1 to FR3, particularly Vκ1 frameworks FR1 to FR3, and a framework FR4, which is selected from a Vλ FR4. [0120] A specific example of a VL having Vκ1 frameworks FR1 to FR4 is represented by SEQ ID NO: 158 (Table 2, FR regions are marked in non-bold). Suitable Vκ1 frameworks FR1 to FR3 with an exemplary Vλ FR4 are set forth in SEQ ID NOs: 159 and 160 (Table 2, 119915P877PC 23.12.2024 Numab Therapeutics AG FR regions are marked in non-bold). Alternative examples of Vκ1 sequences, and examples of Vκ2, Vκ3 or Vκ4 sequences, may be found in Knappik et al., J. Mol. Biol.296 (2000) 57- 86. Suitable Vκ1 frameworks FR1 to FR43 or FR1 to FR4 also comprise, independently of each other, the amino acid sequences having at least 70, 80, 90 percent identity to each of the amino acid sequences corresponding to FR1 to FR3 or FR1 to FR4 of SEQ ID NO: 158 (Table 2, FR regions are marked in non-bold). Suitable Vλ FR4 are as set forth in SEQ ID NO: 161 to SEQ ID NO: 168 and in SEQ ID NO: 169 comprising a single cysteine residue, particular in a case where a second single cysteine is present in the corresponding VH chain, particularly in position 51 (AHo numbering) of VH, for the formation of an inter-domain disulfide bond. In one embodiment, the VL domains of the binding domains of the multispecific antibodies of the invention, when being in Fv or scFv-format, comprises Vλ FR4 comprising the amino acid sequence having at least 80, particularly 90, percent identity to an amino acid sequence selected from any of SEQ ID NO: 161 to SEQ ID NO: 169, or in other words having 1 or 2substitutions when compared to the closest amino acid sequence selected from any of SEQ ID NO: 161 to SEQ ID NO: 169. Particularly, the multispecific antibodies of the invention, when being in Fv or scFv-format, comprises Vλ FR4 comprising an amino acid sequence selected from SEQ ID NOs: 161 or 169. [0121] The binding domains of the multispecific antibodies of the invention comprise a VH domain listed in Tables 1 and 2. Suitably, the binding domains used the invention comprise a VH amino acid sequence listed in Tables 1 and 2, wherein no more than 20 amino acids in the framework sequences (i. e., the sequences which are not CDR sequences) have been mutated (wherein a mutation is, as various non-limiting examples, an addition, substitution or deletion). Suitably, the binding domains of the multispecific antibodies of the invention comprise a VH amino acid sequence listed in one of Tables 1 and 2, wherein no more than 15 amino acids, particularly no more than 10 amino acids, particularly no more than 5 amino acids in the framework sequences (i. e., the sequences which are not CDR sequences) have been mutated (wherein a mutation is, as various non-limiting examples, an addition, substitution or deletion). Other binding domains used in the invention include amino acids that have been mutated, yet have at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identity in the VH regions with the VH regions depicted in the corresponding sequences described in Tables 1 and 2, including VH domains comprising at least positions 5 to 140 (AHo numbering), particularly at least positions 3 to 145 of one of the sequences shown in Tables 1 and 2. [0122] In particular, the binding domains of the multispecific antibodies of the invention comprise a VL domain listed in Tables 1 and 2. Suitably, the binding domains of the multispecific antibodies of the invention comprise a VL amino acid sequence listed in Tables 1 and 2, wherein no more than about 20 amino acids in the framework sequences (i. e., the 119915P877PC 23.12.2024 Numab Therapeutics AG sequences which are not CDR sequences) have been mutated (wherein a mutation is, as various non-limiting examples, an addition, substitution or deletion). Suitably, the binding domains of the multispecific antibodies of the invention comprise a VL amino acid sequence listed in Tables 1 and 2, wherein no more than about 15 amino acids, particularly no more than 10 amino acids, particularly no more than 5 amino acids in the framework sequences (i. e., the sequences which are not CDR sequences) have been mutated (wherein a mutation is, as various non-limiting examples, an addition, substitution or deletion). Other binding domains used in the invention include amino acids that have been mutated, yet have at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identity in the VL regions with a VL region depicted in the sequences described in Tables 1 and 2, including VL domains comprising at least positions 5 to 140 (AHo numbering), particularly at least positions 3 to 145 of one of the sequences shown in Tables 1 and 2. [0123] In the context of the present invention, the term “binding domain used in the present invention” relates to a binding domain as such, i. e. independent of a multispecific context, and, in particular, to a binding domain comprised in a multispecific construct, e. g. one of the binding domains comprised in a bispecific, trispecific or tetraspecific construct. [0124] Specific but non-limiting examples of the anti-CD47xPDL1 multispecific antibodies of the invention are PRO4673, PRO4673_without_dummy_LILRB2-BD, PRO3790, PRO4688, PRO4688_without_dummy_LILRB2-BD, PRO5053, PRO5053_without_dummy_LILRB2-BD, PRO5056 and PRO5056_without_dummy_LILRB2- BD, whose sequences are listed in Tables 3 and 4. [0125] In some embodiments, the multispecific antibody additionally comprises one further binding domain, which has specificity for an antigen different from CD47 and PDL1, wherein said one further binding domain is attached to one of the C-termini of the heavy chains of the immunoglobulin Fc region. [0126] In other embodiments, the multispecific antibody additionally comprises two further binding domains, which have specificity for an antigen different from CD47 and PDL1, wherein said two further binding domains are attached to each of the C-termini of the heavy chains of the immunoglobulin Fc region. [0127] In specific embodiments, the one or two further binding domains specifically bind to a tumor associated antigen (TAA) different from CD47 and PDL1. Suitable TAAs are those known in the art. Examples of suitable TAAs are: ADRB3, AFP, ALK, BCMA, beta human chorionic gonadotropin, CA-125 (MUC16), CAIX, CD123, CD133, CD135, CD135 (FLT3), CD138, CD171, CD19, CD20, CD22, CD24, CD276, CD33, CD33, CD38, CD44v6, CD79b, CD97, CDH3 (cadherin 3), CEA, CEACAM6, CLDN6, CLEC12A (CLL1), CSPG4, CYP1B1, EGFR, EGFRvlll, EPCAM, EPHA2, Ephrin B2, ERBBs (e. g. ERBB2), FAP, FGFR1, folate receptor alpha, folate receptor beta, Fos-related antigen, GA733, GD2, GD3, GFRalpha4, 119915P877PC 23.12.2024 Numab Therapeutics AG globoH, GPC3, GPR20, GPRC5D, HAVCR1, Her2/neu (HER2), HLA-A2, HMWMAA, HPV E6 or E7, human telomerase reverse transcriptase, IL-11Ra, IL-13Ra2, intestinal carboxyl esterase, KIT, Legumain, LewisY, LMP2, Ly6k, MAD-CT-1, MAD-CT-2, ML-IAP, MN-CA IX, MSLN, MUC1, mut hsp 70-2, NA-17, NCAM, neutrophil elastase, NY-BR-1, NY-ESO-1, o- acetyl-GD2, OR51E2, PANX3, PDGFR-beta, PLAC1, Polysialic acid, PSCA, PSMA, RAGE1, ROR1, sLe, sperm protein 17, SSEA-4, SSTR2, sTn antigen, sTn-O-Glycopeptides, TAG72, TARP, TEM1/CD248, TEM7R, thyroglobulin, Tn antigen, Tn-O-Glycopeptides, TPBG (5T4), TRP-2, TSHR, UPK2 and VEGFR2. [0128] In other specific embodiments, the one or two further binding domains specifically bind to an immunomodulatory antigen different from PDL1. Suitable immunomodulatory antigen different from PDL1 are those known in the art. Suitable immunomodulatory antigen different from PDL1 are for example selected from B7-1 (CD80), B7-2 (CD86), B7-DC (CD273), B7-H1 (CD274), B7-H2 (CD275), B7-H3 (CD276), B7-H4 (VTCN1), B7-H5 (VISTA), BTLA (CD272), 4-1BB (CD137), CD137L, CD24, CD27, CD28, CD38, CD40, CD40L (CD154), CD54, CD59, CD70, CTLA-1, CTLA-4 (CD152), CXCL9, GITR (CD357), GITR, HVEM (CD270), ICAM-1 (CD54), ICOS (CD278), LAG-3 (CD223), OX40 (CD134), OX40L (CD252), PD-1 (CD279), TIGIT, CD314, CD334, CD335, CD337, VISTA and TIM-3 (CD366). [0129] In other specific embodiments, the one or two further binding domains specifically bind to an NK cell receptor. Suitable NK cell receptors are those known in the art. Suitable NK cell receptors are for example selected from KIR2DL1, KIR2DL2/3, KIR2DL4, KIR2DL5A, KIR2DL5B, KIR3DL1, KIR3DL2, KIR3DL3, KIR2DS4, KIR3DS, NKG2A, NKG2C, NKG2D, NKG2E, NKp30, NKp44, NKp46, CD2, CD16, CD69, DNAX accessory molecule-1 (DNAM-1), 2B4, NK1.1; a killer immunoglobulin (Ig)-like activating receptors (KAR); ILTs/LIRs (CD85) such as LILRA1, LILRA2, LILRA3, LILRA4, LILRA5, LILRA6, LILRB1, LILRB2, LILRB3, LILRB4, LILRB5, ILT8, ILT10, ILT11 ; NKRP-1; CD94/NKG2A, CD94/NKG2C and CD94/NKG2E heterodimers. [0130] In more specific embodiments, the one or two further binding domains are independently of each other selected from NKp46, LILRB1, LILRB2, LILRB3, LILRB4 and LILRB5, particularly LILRB1, LILRB2, and LILRB4, particularly LILRB1 and LILRB2, particularly LILRB2. [0131] In particular embodiments, the multispecific antibody comprises one further binding domain, which specifically binds to LILRB2 (LILRB2-BDs), wherein said LILRB2-BD is attached to one of the C-termini of the heavy chains of the immunoglobulin Fc region. [0132] In other particular embodiments, the multispecific antibody comprises two further binding domains, which specifically bind to LILRB2 (LILRB2-BDs), wherein said two LILRB2- BDs are attached to each of the C-termini of the heavy chains of the immunoglobulin Fc 119915P877PC 23.12.2024 Numab Therapeutics AG region. The two LILRB2-BD either bind the same or different epitopes on the extracellular domain of LILRB2. Particularly, the two LILRB2-BDs are identical. [0133] The term “same epitope”, as used herein, refers to an individual protein determinant on the protein capable of specific binding to more than one antibody, where that individual protein determinant is identical, i. e. consist of identical chemically active surface groupings of molecules such as amino acids or sugar side chains having identical three-dimensional structural characteristics, as well as identical charge characteristics for each of said antibodies. The term “different epitope”, as used herein in connection with a specific protein target, refers to individual protein determinants on the protein, each capable of specific binding to a different antibody, where these individual protein determinants are not identical for the different antibodies, i. e. consist of non-identical chemically active surface groupings of molecules such as amino acids or sugar side chains having different three-dimensional structural characteristics, as well as different charge characteristics. These different epitopes can be overlapping or non-overlapping. [0134] The term “LILRB2”, i. e. leukocyte immunoglobulin-like receptor subfamily B member 2, also known as immunoglobulin like transcript 4 (ILT4), refers in particular to human LILRB2 with UniProt ID number A2IXV5. LILRB2 belongs to the subfamily B class of LIR receptors and is expressed on immune cells, such as myeloid cells, including monocytes, macrophages, dendritic cells and granulocytes. It binds classical and non- classical major histocompatibility complex Class I (MHC-I) proteins on antigen-presenting cells, such as human leukocyte antigen A (HLA-A) and human leukocyte antigen G (HLA-G), and transduces a negative signal that has immunosuppressive effects. It further binds to several angiopoietin-like proteins (ANGPTLs), such as ANGPTL2 that promotes adaptive inflammation in tissue. A wide range of tumors express HLA-G, a nonclassical MHC-I molecule, which has been shown to mediate immunotolerance in cancer through the interaction with LILRB2, and thus allows such tumor cells to escape innate and adaptive immune response. Blocking of LILRB2 would hinder this escape mechanism. Besides, the blocking of LILRB2 can shift suppressed macrophages (M2) to activated state (M1-like). [0135] There are several anti-LILRB2 antibodies known in the art. However, the number of LILRB2-BDs in the form of antibody fragments, such as scFvs, that can be used as universal building blocks for the construction of multispecific antibodies is limited. It is thus desirable to have suitable anti-LILRB2 antibody building blocks at hand that can readily be incorporated into any desirable multispecific antibody format, and that are capable of potently blocking the biological function of LILRB2. Furthermore, said building blocks should have superior biophysical properties, such as in particular a high stability, in order to facilitate their efficient incorporation into multispecific antibodies. 119915P877PC 23.12.2024 Numab Therapeutics AG [0136] Suitably, the LIRB2-BDs of the multispecific antibodies of the invention target in particular human LILRB2. Particularly, the LIRB2-BDs of the multispecific antibodies of the invention target human and cynomolgus (Macaca fascicularis) LILRB2. [0137] Suitably, the LILRB2-BDs used in the present invention are characterized by one or more of the following parameters: a. when in scFv format, bind to human LILRB2 with a monovalent dissociation constant (KD) of 10 pM to 5 nM, particularly with a KD of 20 pM to 3 nM, particularly of 50 pM to 1 nM, as measured by surface plasmon resonance (SPR); b. block the interaction of LILRB2 to its ligand HLA-G with an IC50 of 100 pM to 10 nM, particularly of 100 pM to 5 nM, particularly of 100 pM to 2 nM, as measured by flow cytometry; c. block the interaction of LILRB2 to its ligand HLA-A3 with an IC50 of 100 pM to 10 nM, particularly of 100 pM to 5 nM, particularly of 100 pM to 2 nM, as measured by flow cytometry; d. are not cross-reactive with members of leukocyte immunoglobulin-like receptor subfamily A (LILRA); and/or e. are not cross reactive with members of leukocyte immunoglobulin-like receptor subfamily LILRB3, LILRB4 and LILRB5. [0138] The expression “members of leukocyte immunoglobulin-like receptor subfamily A” or “LILRA” refers to leukocyte immunoglobulin-like receptor subfamily A, which, like the LILRBs, are a family of immunoreceptors that belong to leukocyte Ig-like receptors (LIRs) and that are expressed predominantly on monocytes and B cells and at lower levels on dendritic cells and natural killer (NK) cells. Examples are LILRA1, LILRA2, LILRA3, LILRA4, LILRA5 and LILRA6. [0139] The expression “LILRB members other than LILRB2” refer to members of leukocyte immunoglobulin-like receptor subfamily B different from LILRB2. Examples are LILRB1, LILRB3, LILRB4 and LILRB5. [0140] Suitably, the LILRB2-BDs of the multispecific antibodies of the present invention are further characterized by one or more of the following parameters: a. when in scFv format, have a melting temperature (Tm), determined by differential scanning fluorimetry, in the range of 55°C to 85°C, particularly in the range of 60°C to 85°C, in particular wherein said antibody is formulated in 50 mM phosphate-citrate buffer at pH 6.4, 150 mM NaCl; b. when in scFv format, have a loss in monomer content, after storage for four weeks, at ‑80°C, of less than 5 %, less than 4 %, less than 3 %, less than 2 %, particularly less than 1.5 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, 119915P877PC 23.12.2024 Numab Therapeutics AG and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4; c. when in scFv format, have a loss in monomer content, after storage for four weeks, at 4°C, of less than 15 %, e. g. less than 12 %, less than 10 %, less than 9 %, less than 8 %, particularly less than 7 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4; d. when in scFv format, have a loss in monomer content, after storage for four weeks, at 40°C, of less than 15 %, e. g. less than 12 %, less than 10 %, less than 9 %, less than 8 %, particularly less than 7 %, when the antibody of the invention is at a starting concentration of 10 mg/ml, and in particular wherein said antibody is formulated in 50 mM phosphate citrate buffer with 150 mM NaCl at pH 6.4. [0141] Suitably, the LILRB2-BDs of the multispecific antibodies of the invention are binding domains provided in the present disclosure. The LILRB2-BDs of the multispecific antibodies of the invention include, but are not limited to, the humanized LILRB2-BDs whose sequences are listed in Table 1. [0142] In a particular aspect, the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequences of SEQ ID NO: 7; or ii. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequences of SEQ ID NO: 7; b) one binding domain, which specifically binds to PDL1 (PDL1-BD); wherein said PDL1-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 21, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, 119915P877PC 23.12.2024 Numab Therapeutics AG the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequences of SEQ ID NO: 27; or ii. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 21, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequences of SEQ ID NO: 27; or iii. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequences of SEQ ID NO: 27; or iv. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequences of SEQ ID NO: 27; or v. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 78, and the LCDR3 sequences of SEQ ID NO: 80; or vi. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 79, and the LCDR3 sequences of SEQ ID NO: 80; c) an IgG-like region comprising an immunoglobulin Fc region, a first Fab arm and a second Fab arm, wherein: - the CD47 BD is an scFv fragment replacing the first Fab arm, and - the PDL1 BD is located in the second Fab arm or is an scFv fragment replacing said second Fab arm; 119915P877PC 23.12.2024 Numab Therapeutics AG d) two binding domains, which specifically bind to LILRB2 (LILRB2-BDs); wherein said LILRB2-BDs comprise: i. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequences of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 97, and the LCDR3 sequences of SEQ ID NO: 99; or ii. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequences of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 98, and the LCDR3 sequences of SEQ ID NO: 99; or iii. the HCDR1 sequence of SEQ ID NO: 112, the HCDR2 sequences of SEQ ID NO: 113, the HCDR3 sequence of SEQ ID NO: 114, the LCDR1 sequence of SEQ ID NO: 115, the LCDR2 sequence of SEQ ID NO: 116, and the LCDR3 sequences of SEQ ID NO: 118; or iv. the HCDR1 sequence of SEQ ID NO: 112, the HCDR2 sequences of SEQ ID NO: 113, the HCDR3 sequence of SEQ ID NO: 114, the LCDR1 sequence of SEQ ID NO: 115, the LCDR2 sequence of SEQ ID NO: 117, and the LCDR3 sequences of SEQ ID NO: 118 or v. the HCDR1 sequence of SEQ ID NO: 128, the HCDR2 sequences of SEQ ID NO: 129, the HCDR3 sequence of SEQ ID NO: 130, the LCDR1 sequence of SEQ ID NO: 131, the LCDR2 sequence of SEQ ID NO: 132, and the LCDR3 sequences of SEQ ID NO: 134; or vi. the HCDR1 sequence of SEQ ID NO: 128, the HCDR2 sequences of SEQ ID NO: 129, the HCDR3 sequence of SEQ ID NO: 130, the LCDR1 sequence of SEQ ID NO: 131, the LCDR2 sequence of SEQ ID NO: 133, and 119915P877PC 23.12.2024 Numab Therapeutics AG the LCDR3 sequences of SEQ ID NO: 134; and wherein said two LILRB2-BDs are attached to each of the C-termini of the heavy chains of the immunoglobulin Fc region. [0143] In a more particular aspect, the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequences of SEQ ID NO: 7; or ii. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequences of SEQ ID NO: 7; b) one binding domain, which specifically binds to PDL1 (PDL1-BD); wherein said PDL1-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 78, and the LCDR3 sequences of SEQ ID NO: 80; or ii. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 79, and the LCDR3 sequences of SEQ ID NO: 80; c) an IgG-like region comprising an immunoglobulin Fc region, a first Fab arm and a second Fab arm, wherein: - the CD47 BD is an scFv fragment replacing the first Fab arm, and - the PDL1 BD is located in the second Fab arm or is an scFv fragment replacing said second Fab arm; 119915P877PC 23.12.2024 Numab Therapeutics AG d) two binding domains, which specifically bind to LILRB2 (LILRB2-BDs), wherein said two LILRB2-BDs are attached to each of the C-termini of the heavy chains of the immunoglobulin Fc region. [0144] In another more particular aspect, the present invention relates to a multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD); wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequences of SEQ ID NO: 7; or ii. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequences of SEQ ID NO: 7; b) one binding domain, which specifically binds to PDL1 (PDL1-BD); wherein said PDL1-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 78, and the LCDR3 sequences of SEQ ID NO: 80; or ii. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 79, and the LCDR3 sequences of SEQ ID NO: 80; c) an IgG-like region comprising an immunoglobulin Fc region, a first Fab arm and a second Fab arm, wherein: - the CD47 BD is an scFv fragment replacing the first Fab arm, and - the PDL1 BD is located in the second Fab arm or is an scFv fragment replacing said second Fab arm; 119915P877PC 23.12.2024 Numab Therapeutics AG d) two binding domains, which specifically bind to LILRB2 (LILRB2-BDs); wherein said LILRB2-BDs comprise: i. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequences of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 97, and the LCDR3 sequences of SEQ ID NO: 99; or ii. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequences of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 98, and the LCDR3 sequences of SEQ ID NO: 99; and wherein said two LILRB2-BDs are attached to each of the C-termini of the heavy chains of the immunoglobulin Fc region. [0145] Suitably, the binding domains of the multispecific antibodies of the invention, except for the CD47-BD, are selected from the group consisting of: a Fab, an scFab, an Fv, a dsFv, an scFv and a dsscFv, particularly from a Fab, Fv and scFv. [0146] Suitably, the binding domains of the multispecific antibodies of the invention are operably linked. The binding domains of the multispecific antibodies of the invention are capable of binding to their respective antigens or receptors simultaneously. The term “simultaneously”, as used herein refers for example to the simultaneous binding of the CD47-BD and the PDL1-BD or the simultaneous binding of the PDL1-BD and the LILRB2- BD. [0147] The term “operably linked”, as used herein, indicates that two molecules (e. g., polypeptides, domains, binding domains) are attached in a way that each molecule retains functional activity. Two molecules can be “operably linked” whether they are attached directly or indirectly (e. g., via a linker, via a moiety, via a linker to a moiety). [0148] The term “linker” refers to a peptide or other moiety that is optionally located between binding domains or antibody fragments used in the invention. A number of strategies may be used to covalently link molecules together. These include, but are not limited to, polypeptide linkages between N- and C-termini of proteins or protein domains, linkage via disulfide bonds, and linkage via chemical cross-linking reagents. In one aspect of this embodiment, the linker is a peptide bond, generated by recombinant techniques or peptide synthesis. Choosing a suitable linker for a specific case where two polypeptide chains are to be connected depends on various parameters, including but not limited to the 119915P877PC 23.12.2024 Numab Therapeutics AG nature of the two polypeptide chains (e. g., whether they naturally oligomerize), the distance between the N- and the C-termini to be connected if known, and/or the stability of the linker towards proteolysis and oxidation. Furthermore, the linker may contain amino acid residues that provide flexibility. [0149] In the context of the present invention, the term “polypeptide linker” refers to a linker consisting of a chain of amino acid residues linked by peptide bonds that is connecting two domains, each being attached to one end of the linker. The polypeptide linker should have a length that is adequate to link two molecules in such a way that they assume the correct conformation relative to one another so that they retain the desired activity. In particular embodiments, the polypeptide linker has a continuous chain of between 2 and 30 amino acid residues (e. g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 amino acid residues). In addition, the amino acid residues selected for inclusion in the polypeptide linker should exhibit properties that do not interfere significantly with the activity of the polypeptide. Thus, the linker peptide on the whole should not exhibit a charge that would be inconsistent with the activity of the polypeptide, or interfere with internal folding, or form bonds or other interactions with amino acid residues in one or more of the monomers that would seriously impede the binding of receptor monomer domains. In particular embodiments, the polypeptide linker is non-structured polypeptide. Useful linkers include glycine-serine, or GS linkers. By “Gly-Ser” or “GS” linkers is meant a polymer of glycines and serines in series (including, for example, (Gly-Ser)n, (GSGGS)n (SEQ ID NO: 259), (GGGGS)n (SEQ ID NO: 260) and (GGGS)n (SEQ ID NO: 261), where n is an integer of at least one), glycine-alanine polymers, alanine-serine polymers, and other flexible linkers such as the tether for the shaker potassium channel, and a large variety of other flexible linkers, as will be appreciated by those in the art. Glycine-serine polymers are preferred since oligopeptides comprising these amino acids are relatively unstructured, and therefore may be able to serve as a neutral tether between components. Secondly, serine is hydrophilic and therefore able to solubilize what could be a globular glycine chain. Third, similar chains have been shown to be effective in joining subunits of recombinant proteins such as single-chain antibodies. [0150] In particular embodiments the one or two LILRB2-BD(s) is/are fused to the heavy chain of the immunoglobulin Fc region via a linker L1. [0151] The linker L1 is a peptide of 2-30 amino acids, more particularly 5-25 amino acids, and most particularly 10-20 amino acids. In particular embodiments, said linker L1 comprises one or more units of four (4) glycine amino acid residues and one (1) serine amino acid residue (GGGGS)n (SEQ ID NO:170), wherein n=1, 2, 3, 4 or 5, particularly n=2. 119915P877PC 23.12.2024 Numab Therapeutics AG [0152] The scFv domains of the multispecific antibodies of the invention comprise a variable heavy chain domain (VH) and a variable light chain domain (VL) connected by a linker L2. [0153] The linker L2 is a peptide of 10-40 amino acids, more particularly 15-30 amino acids, and most particularly 20-25 amino acids. In particular embodiments, said linker L2 comprises one or more units of four (4) glycine amino acid residues and one (1) serine amino acid residue (GGGGS)n (SEQ ID NO:172), wherein n=1, 2, 3, 4, 5, 6, 7 or 8, particularly n=4. [0154] Specific but non-limiting examples of the anti-CD47xPDL1xLILRB2 multispecific antibodies of the invention are PRO4672, PRO4687, PRO5052 and PRO5055, whose sequences are listed in table 4. [0155] The multispecific antibodies of the invention can be produced using any convenient antibody-manufacturing method known in the art (see, e. g., Fischer, N. & Leger, O., Pathobiology 74 (2007) 3-14 with regard to the production of bispecific constructs; Hornig, N. & Färber-Schwarz, A., Methods Mol. Biol.907 (2012)713-727, and WO 99/57150 with regard to bispecific diabodies and tandem scFvs). Specific examples of suitable methods for the preparation of the bispecific construct further include, inter alia, the Genmab (see Labrijn et al., Proc. Natl. Acad. Sci. USA 110 (2013) 5145-5150) and Merus (see de Kruif et al., Biotechnol. Bioeng.106 (2010) 741-750) technologies. Methods for production of bispecific antibodies comprising a functional antibody Fc part are also known in the art (see, e. g., Zhu et al., Cancer Lett.86 (1994) 127-134); and Suresh et al., Methods Enzymol.121 (1986) 210-228). [0156] These methods typically involve the generation of monoclonal antibodies, for example by means of fusing myeloma cells with the spleen cells from a mouse that has been immunized with the desired antigen using the hybridoma technology (see, e. g., Yokoyama et al., Curr. Protoc. Immunol. Chapter 2, Unit 2.5, 2006) or by means of recombinant antibody engineering (repertoire cloning or phage display/yeast display) (see, e. g., Chames & Baty, FEMS Microbiol. Letters 189 (2000) 1-8), and the combination of the antigen-binding domains or fragments or parts thereof of two or more different monoclonal antibodies to give a bispecific or multispecific construct using known molecular cloning techniques. [0157] The multispecific antibodies of the invention can be prepared by conjugating the constituent binding specificities, using methods known in the art. For example, each binding specificity of the bispecific molecule can be generated separately and then conjugated to one another. When the binding specificities are proteins or peptides, a variety of coupling or cross-linking agents can be used for covalent conjugation. Examples of cross-linking agents include protein A, carbodiimide, N-succinimidyl-5-acetyl-thioacetate (SATA), 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-succinimidyl-3-(2- 119915P877PC 23.12.2024 Numab Therapeutics AG pyridyldithio)propionate (SPDP), and sulfosuccinimidyl 4-(N-maleimidomethyl)-cyclohexane- l-carboxylate (sulfo-SMCC) (see e. g., Karpovsky et al., 1984 J. Exp. Med.160: 1686; Liu, M A et al., 1985 Proc. Natl. Acad. Sci. USA 82:8648). Other methods include those described in Paulus, 1985 Behring Ins. Mitt. No.78, 118-132; Brennan et al., 1985 Science 229:81-83, and Glennie et al., 1987 J. Immunol.139: 2367-2375. Conjugating agents are SATA and sulfo-SMCC, both available from Pierce Chemical Co. (Rockford, III). [0158] When the binding specificities are antibodies, they can be conjugated by sulfhydryl bonding of the C-terminus hinge regions of the two heavy chains. In a particular embodiment, the hinge region is modified to contain an odd number of sulfhydryl residues, for example one, prior to conjugation. [0159] Alternatively, two or more binding specificities can be encoded in the same vector and expressed and assembled in the same host cell. This method is particularly useful where the bispecific molecule is a mAb x Fab, a mAb x scFv, a mAb x dsFv or a mAb x Fv fusion protein. Methods for preparing multispecific antibodies and molecules are described for example in U.S. Pat. No.5,260,203; U.S. Pat. No.5,455,030; U.S. Pat. No.4,881,175; U.S. Pat. No.5,132,405; U.S. Pat. No.5,091,513; U.S. Pat. No.5,476,786; U.S. Pat. No. 5,013,653; U.S. Pat. No.5,258,498; and U.S. Pat. No.5,482,858. [0160] Binding of the multispecific antibodies to their specific targets can be confirmed by, for example, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (REA), FACS analysis, bioassay (e. g., growth inhibition), or Western Blot assay. Each of these assays generally detects the presence of protein-antibody complexes of particular interest by employing a labeled reagent (e. g., an antibody) specific for the complex of interest. [0161] In a further aspect, the invention provides a nucleic acid or two nucleic acids or three nucleic acids encoding the multispecific antibody of the invention. Such nucleic acids can be optimized for expression in mammalian cells. [0162] The term “nucleic acid” is used herein interchangeably with the term “polynucleotide(s)” and refers to one or more deoxyribonucleotides or ribonucleotides and polymers thereof in either single- or double-stranded form. The term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphorates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs). Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e. g., degenerate codon substitutions) and complementary sequences, as well as the sequence explicitly indicated. Specifically, as detailed below, degenerate codon 119915P877PC 23.12.2024 Numab Therapeutics AG substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues (Batzer et al., Nucleic Acid Res.19:5081, 1991; Ohtsuka et al., J. Biol. Chem.260:2605- 2608, 1985; and Rossolini et al., Mol. Cell. Probes 8:91-98, 1994). [0163] The invention provides substantially purified nucleic acid molecules which encode polypeptides comprising segments or domains of the multispecific antibody described above. When expressed from appropriate expression vectors, polypeptides encoded by these nucleic acid molecules are capable of exhibiting antigen-binding capacities of the multispecific antibody of the present invention. [0164] The polynucleotide sequences can be produced by de novo solid-phase DNA synthesis or by PCR mutagenesis of an existing sequence (e. g., sequences as described in the Examples below) encoding the multispecific antibody of the invention or fragments thereof or binding domains thereof. Direct chemical synthesis of nucleic acids can be accomplished by methods known in the art, such as the phosphotriester method of Narang et al., 1979, Meth. Enzymol.68:90; the phosphodiester method of Brown et al., Meth. Enzymol.68: 109, 1979; the diethylphosphoramidite method of Beaucage et al., Tetra. Lett., 22: 1859, 1981; and the solid support method of U.S. Pat. No.4,458,066. Introducing mutations to a polynucleotide sequence by PCR can be performed as described in, e. g., PCR Technology: Principles and Applications for DNA Amplification, H. A. Erlich (Ed.), Freeman Press, NY, N.Y., 1992; PCR Protocols: A Guide to Methods and Applications, Innis et al. (Ed.), Academic Press, San Diego, Calif, 1990; Mattila et al., Nucleic Acids Res. 19:967, 1991; and Eckert et al., PCR Methods and Applications 1:17, 1991. [0165] Also provided in the invention are expression vectors and host cells for producing the multispecific antibody of the invention or fragments thereof or binding domains thereof. [0166] The term “vector” is intended to refer to a polynucleotide molecule capable of transporting another polynucleotide to which it has been linked. One type of vector is a “plasmid”, which refers to a circular double stranded DNA loop into which additional DNA segments may be ligated. Another type of vector is a viral vector, wherein additional DNA segments may be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e. g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e. g., non- episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. [0167] Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as “recombinant expression vectors” (or simply, “expression vectors”). In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. In the present specification, 119915P877PC 23.12.2024 Numab Therapeutics AG “plasmid” and “vector” may be used interchangeably as the plasmid is the most commonly used form of vector. However, the invention is intended to include such other forms of expression vectors, such as viral vectors (e. g., replication defective retroviruses, adenoviruses and adeno- associated viruses), which serve equivalent functions. In this particular context, the term “operably linked” refers to a functional relationship between two or more polynucleotide (e. g., DNA) segments. Typically, it refers to the functional relationship of a transcriptional regulatory sequence to a transcribed sequence. For example, a promoter or enhancer sequence is operably linked to a coding sequence if it stimulates or modulates the transcription of the coding sequence in an appropriate host cell or other expression system. Generally, promoter transcriptional regulatory sequences that are operably linked to a transcribed sequence are physically contiguous to the transcribed sequence, i. e., they are cis-acting. However, some transcriptional regulatory sequences, such as enhancers, need not be physically contiguous or located in close proximity to the coding sequences whose transcription they enhance. [0168] Various expression vectors can be employed to express the polynucleotides encoding the multispecific antibody chains. Both viral-based and non-viral expression vectors can be used to produce the antibodies in a mammalian host cell. Non-viral vectors and systems include plasmids, episomal vectors, typically with an expression cassette for expressing a protein or RNA, and human artificial chromosomes (see, e. g., Harrington et al., Nat Genet.15:345, 1997). For example, non-viral vectors useful for expression of the polynucleotides encoding the multispecific antibody chains in mammalian (e. g., human) cells include pThioHis A, B and C, pcDNA3.1/His, pEBVHis A, B and C, (Invitrogen, San Diego, Calif.), MPS V vectors, and numerous other vectors known in the art for expressing other proteins. Useful viral vectors include vectors based on retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, vectors based on SV40, papilloma virus, HBP Epstein Barr virus, vaccinia virus vectors and Semliki Forest virus (SFV). See, Brent et al., supra; Smith, Annu. Rev. Microbiol.49:807, 1995; and Rosenfeld et al., Cell 68: 143, 1992. [0169] The choice of expression vector depends on the intended host cells in which the vector is to be expressed. Typically, the expression vectors contain a promoter and other regulatory sequences (e. g., enhancers) that are operably linked to the polynucleotides encoding a multispecific antibody chain or a fragment. In one embodiment, an inducible promoter is employed to prevent expression of inserted sequences except under inducing conditions. Inducible promoters include, e. g., arabinose, lacZ, metallothionein promoter or a heat shock promoter. Cultures of transformed organisms can be expanded under non- inducing conditions without biasing the population for coding sequences whose expression products are better tolerated by the host cells. In addition to promoters, other regulatory elements may also be required or desired for efficient expression of a multispecific antibody 119915P877PC 23.12.2024 Numab Therapeutics AG chain or a fragment. These elements typically include an ATG initiation codon and adjacent ribosome-binding site or other sequences. In addition, the efficiency of expression may be enhanced by the inclusion of enhancers appropriate to the cell system in use (see, e. g., Scharf et al., Results Probl. Cell Differ.20: 125, 1994; and Bittner et al., Meth. Enzymol., 153:516, 1987). For example, the SV40 enhancer or CMV enhancer may be used to increase expression in mammalian host cells. [0170] Vectors to be used typically encode the multispecific antibody light and heavy chain or chains including constant regions or parts thereof. Such vectors allow expression of the variable regions as fusion proteins with the constant regions thereby leading to production of intact antibodies and antigen-binding fragments thereof. Typically, such constant regions are human. [0171] The term “recombinant host cell” (or simply “host cell”) refers to a cell into which a recombinant expression vector has been introduced. It should be understood that such terms are intended to refer not only to the particular subject cell but to the progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term “host cell” as used herein. [0172] The host cells for harboring and expressing the multispecific antibody of the invention can be either prokaryotic or eukaryotic. E. coli is one prokaryotic host useful for cloning and expressing the polynucleotides of the present invention. Other microbial hosts suitable for use include bacilli, such as Bacillus subtilis, and other enterobacteriaceae, such as Salmonella, Serratia, and various Pseudomonas species. In these prokaryotic hosts, one can also make expression vectors, which typically contain expression control sequences compatible with the host cell (e. g., an origin of replication). In addition, any number of a variety of well-known promoters will be present, such as the lactose promoter system, a tryptophan (trp) promoter system, a beta-lactamase promoter system, or a promoter system from phage lambda. The promoters typically control expression, optionally with an operator sequence, and have ribosome-binding site sequences and the like, for initiating and completing transcription and translation. Other microbes, such as yeast, can also be employed to express the multispecific antibodies of the invention. Insect cells in combination with baculovirus vectors can also be used. [0173] In one embodiment, mammalian host cells are used to express and produce the multispecific antibody of the invention. For example, they can be either a hybridoma cell line expressing endogenous immunoglobulin genes or a mammalian cell line harboring an exogenous expression vector. These include any normal mortal or normal or abnormal immortal animal or human cell. For example, a number of suitable host cell lines capable of secreting intact immunoglobulins have been developed including the CHO cell lines, various 119915P877PC 23.12.2024 Numab Therapeutics AG COS cell lines, HeLa cells, myeloma cell lines, transformed B-cells and hybridomas. The use of mammalian tissue cell culture to express polypeptides is discussed generally in, e. g., Winnacker, FROM GENES TO CLONES, VCH Publishers, N.Y., N.Y., 1987. Expression vectors for mammalian host cells can include expression control sequences, such as an origin of replication, a promoter, and an enhancer (see, e. g., Queen, et al., Immunol. Rev. 89:49-68, 1986), and necessary processing information sites, such as ribosome-binding sites, RNA splice sites, polyadenylation sites, and transcriptional terminator sequences. These expression vectors usually contain promoters derived from mammalian genes or from mammalian viruses. Suitable promoters may be constitutive, cell type-specific, stage- specific, and/or modulatable or regulatable. Useful promoters include, but are not limited to, the metallothionein promoter, the constitutive adenovirus major late promoter, the dexamethasone-inducible MMTV promoter, the SV40 promoter, the MRP pol III promoter, the constitutive MPS V promoter, the tetracycline-inducible CMV promoter (such as the human immediate-early CMV promoter), the constitutive CMV promoter, and promoter- enhancer combinations known in the art. [0174] Methods for introducing expression vectors containing the polynucleotide sequences of interest vary depending on the type of cellular host. For example, calcium chloride transfection is commonly utilized for prokaryotic cells, whereas calcium phosphate treatment or electroporation may be used for other cellular hosts. (See generally Green, M. R., and Sambrook, J., Molecular Cloning: A Laboratory Manual (Fourth Edition), Cold Spring Harbor Laboratory Press (2012)). Other methods include, e. g., electroporation, calcium phosphate treatment, liposome-mediated transformation, injection and microinjection, ballistic methods, virosomes, immunoliposomes, polycation-nucleic acid conjugates, naked DNA, artificial virions, fusion to the herpes virus structural protein VP22 (Elliot and O'Hare, Cell 88:223, 1997), agent-enhanced uptake of DNA, and ex vivo transduction. For long-term, high-yield production of recombinant proteins, stable expression will often be desired. For example, cell lines which stably express the multispecific antibody of the invention can be prepared using expression vectors of the invention which contain viral origins of replication or endogenous expression elements and a selectable marker gene. Following the introduction of the vector, cells may be allowed to grow for 1 to 2 days in an enriched media before they are switched to selective media. The purpose of the selectable marker is to confer resistance to selection, and its presence allows growth of cells which successfully express the introduced sequences in selective media. Resistant, stably transfected cells can be proliferated using tissue culture techniques appropriate to the cell type. The present invention thus provides a method of producing the antibody of the invention or antigen- binding fragment thereof, wherein said method comprises the step of culturing a host cell comprising a nucleic acid or a vector encoding the antibody of the invention or antigen- 119915P877PC 23.12.2024 Numab Therapeutics AG binding fragment thereof, whereby said antibody of the disclosure or a fragment thereof is expressed. [0175] In one aspect, the present invention relates to a method of producing the multispecific antibody of the invention, the method comprising the step of culturing a host cell expressing a nucleic acid or two nucleic acids or three nucleic acids encoding the multispecific antibody of the invention. In particular, the present invention relates to a method of producing the multispecific antibody of the invention, the method comprising (i) providing a nucleic acid or two nucleic acids or three nucleic acids encoding the multispecific antibody of the invention or a vector or two vectors or three vectors encoding the multispecific antibody of the invention, expressing said nucleic acid or nucleic acids, or said vector or vectors, and collecting said multispecific antibody from the expression system, or (ii) providing a host cell or host cells expressing a nucleic acid or nucleic acids encoding the multispecific antibody of the invention, culturing said host cell or said host cells; and collecting said multispecific antibody from the cell culture. [0176] In a further aspect, the present invention relates to a pharmaceutical composition comprising the multispecific antibody of the invention, and a pharmaceutically acceptable carrier. "Pharmaceutically acceptable carrier" means a medium or diluent that does not interfere with the structure of the antibodies. Pharmaceutically acceptable carriers enhance or stabilize the composition, or facilitate preparation of the composition. Pharmaceutically acceptable carriers include solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. [0177] Certain, of such carriers enable pharmaceutical compositions to be formulated as, for example, tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspension and lozenges for the oral ingestion by a subject. Certain of such carriers enable pharmaceutical compositions to be formulated for injection, infusion or topical administration. For example, a pharmaceutically acceptable carrier can be a sterile aqueous solution. [0178] The pharmaceutical composition of the invention can be administered by a variety of methods known in the art. The route and/or mode of administration vary depending upon the desired results. Administration can be intravenous, intramuscular, intraperitoneal, or subcutaneous, or administered proximal to the site of the target. In particular embodiments, the administration is intramuscular, or subcutaneous, particularly subcutaneous. The pharmaceutically acceptable carrier should be suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e. g., by injection or infusion), particularly for intramuscular or subcutaneous administration. Depending on the route of administration, the active compound, i. e., the multispecific antibody of the invention, may be 119915P877PC 23.12.2024 Numab Therapeutics AG coated in a material to protect the compound from the action of acids and other natural conditions that may inactivate the compound. [0179] The pharmaceutical compositions of the invention can be prepared in accordance with methods well known and routinely practiced in the art. See, e. g., Remington: The Science and Practice of Pharmacy, Mack Publishing Co., 20th ed., 2000; and Sustained and Controlled Release Drug Delivery Systems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, 1978. Pharmaceutical compositions are preferably manufactured under GMP conditions. Typically, a therapeutically effective dose or efficacious dose of the multispecific antibody of the invention is employed in the pharmaceutical compositions of the invention. The multispecific antibodies of the invention are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those of skill in the art. Dosage regimens are adjusted to provide the optimum desired response (e. g., a therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. [0180] Actual dosage levels of the active ingredients in the pharmaceutical compositions of the invention can be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The selected dosage level depends upon a variety of pharmacokinetic factors including the activity of the particular compositions of the present invention employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors. [0181] The multispecific antibody of the invention is usually administered on multiple occasions. Intervals between single dosages can be weekly, monthly or yearly. Intervals can also be irregular as indicated by measuring blood levels of the multispecific antibody of the invention in the patient. Alternatively, the multispecific antibody of the invention can be administered as a sustained release formulation, in which case less frequent administration is required. Dosage and frequency vary depending on the half-life of the antibody in the 119915P877PC 23.12.2024 Numab Therapeutics AG patient. In general, humanized antibodies show longer half-life than that of chimeric antibodies and nonhuman antibodies. The dosage and frequency of administration can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, a relatively low dosage is administered at relatively infrequent intervals over a long period of time. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, a relatively high dosage at relatively short intervals is sometimes required until progression of the disease is reduced or terminated, and preferably until the patient shows partial or complete amelioration of symptoms of disease. Thereafter, the patient can be administered a prophylactic regime. [0182] In certain embodiments, the pharmaceutical composition comprises: a) a first multispecific antibody that specifically binds to CD47 and PDL1; b) a second antibody that specifically binds to LILRB2; and c) a pharmaceutically acceptable carrier. [0183] In particular embodiments, the first multispecific antibody that specifically binds to CD47 and PDL1 is selected from anti-CD47xPDL1 antibodies that are currently tested or have been tested in clinical trials. [0184] The expression “antibodies that have been tested in clinical trials” refers to antibodies that have successfully passed clinical testing and also to antibodies that failed in clinical testing. The expression “antibodies that are currently tested in clinical trials” refers to antibodies that entered clinical trials but have not yet passed clinical trials or have not been withdrawn from clinical trials. [0185] Specific non limiting examples of anti-CD47xPDL1 antibodies that are currently tested or have been tested in clinical trials are: IBI-322 by Innovent Biologics Inc.; PF- 07257876 by Pfizer Inc.; 6MW-3211 by Mabwell (Shanghai) Bioscience Co. Ltd.; BAT7104 by Bio-Thera Solutions, Ltd.; QL401 by QLSF Biotherapeutics, Inc.; LB-101 by Centessa Pharmaceuticals plc; Imm2520 by ImmuneOnco Biopharmaceuticals Shanghai Inc.; SHS- 009 by Nanjing Sanhome Pharmaceutical Co Ltd.; BCD-106 by Biocad Biopharmaceutical Co.; TJL-1C4 by I-MAB Biopharma Co. Ltd.; SG12473 by Hangzhou Sumgen Biotech Co. Ltd.; XB014 by Exelixis Inc.; PMC-122 by PharmAbcine Inc.; ABP-160 by Abpro Corp.; papiliximab by Shaperon Inc.; JY-207 by Shenzhen Enduring Biotech Ltd.; NY2601 and NY2901 by Light Chain Bioscience AG. [0186] In other particular embodiments, the first multispecific antibody that specifically binds to CD47 and PDL1 is selected from an anti-CD47xPDL1 multispecific antibody according to the invention, as defined herein. [0187] In yet other particular embodiments, the second antibody is a fragment-based antibody further comprising one hSA-binding domain. 119915P877PC 23.12.2024 Numab Therapeutics AG [0188] In further particular embodiments, the second antibody that specifically binds to LILRB2 is selected from anti-LILRB2 antibodies that are currently tested or have been tested in clinical trials. [0189] Specific non limiting examples of anti-LILRB2 antibodies that are currently tested or have been tested in clinical trials are: JTX-8064 by Jounce Therapeutics Inc.; MK-4830 by Merck & Co. Inc.; IO-108 by Immune-Onc Therapeutics Inc.; NGM-707 by NGM Biopharmaceuticals Inc.; BMS-986406 by Bristol Myers Squibb Company; IOS-1002 by ImmunOs Therapeutics AG; CDX-585 by Celldex Therapeutics Inc.; SPX-303 by SparX Biopharmaceutical Corp.; CHS-1000 by Coherus BioSciences Inc.; ES009 by Elpiscience (Suzhou) Biopharma Ltd.; XBH41 by GV20 Therapeutics and IVS-5001 by Invectys. [0190] In other particular embodiments, the second antibody that specifically binds to LILRB2 is selected from antibodies that comprise at least one LILRB2-BD as defined herein. [0191] In yet other particular embodiments, the second antibody that specifically binds to LILRB2 is selected from antibodies that comprise two LILRB2-BD as defined herein. Specific non-limiting examples of such anti-LILRB2 antibodies are PRO3603, PRO3635 and PRO4710, whose sequences are listed in Table 5. [0192] In one aspect, the present invention relates to the multispecific antibody of the invention or the pharmaceutical composition of the invention for use as a medicament. In a suitable embodiment, the present invention provides the multispecific antibody or the pharmaceutical composition for use in the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer. [0193] In another aspect, the present invention provides the pharmaceutical composition for use in the manufacture of a medicament for the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer. [0194] In another aspect, the present invention relates to the use of the multispecific antibody or the pharmaceutical composition for treating a disease, particularly a human disease, more particularly a human disease selected from cancer, in a subject in need thereof. [0195] In another aspect, the present invention relates to a method of treating a subject comprising administering to the subject a therapeutically effective amount of the multispecific antibody of the present invention. In a suitable embodiment, the present invention relates to a method for the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer, in a subject comprising administering to the subject a therapeutically effective amount of the multispecific antibody of the present invention. [0196] The term “subject” includes human and non-human animals. [0197] The term “animals” include all vertebrates, e. g., non-human mammals and non- mammals, such as non-human primates, sheep, dog, cow, chickens, amphibians, and 119915P877PC 23.12.2024 Numab Therapeutics AG reptiles. Except when noted, the terms “patient” or “subject” are used herein interchangeably. [0198] The terms “treatment”, “treating”, “treat”, “treated”, and the like, as used herein, refer to obtaining a desired pharmacologic and/or physiologic effect. The effect may be therapeutic in terms of a partial or complete cure for a disease and/or adverse effect attributable to the disease or delaying the disease progression. “Treatment”, as used herein, covers any treatment of a disease in a mammal, e. g., in a human, and includes: (a) inhibiting the disease, i. e., arresting its development; and (b) relieving the disease, i. e., causing regression of the disease. [0199] The term “therapeutically effective amount” or “efficacious amount” refers to the amount of an agent that, when administered to a mammal or other subject for treating a disease, is sufficient to affect such treatment for the disease. The “therapeutically effective amount” will vary depending on the agent, the disease and its severity and the age, weight, etc., of the subject to be treated. [0200] In one embodiment, the disease is a cancer selected from: a benign or especially malignant tumor, solid tumors, mesothelioma, brain cancer, kidney cancer, liver cancer, adrenal gland cancer, bladder cancer, breast cancer, stomach cancer (e. g., gastric tumors), esophageal cancer, ovarian cancer, cervical cancer, thymic cancer, choriocarcinoma, oral cancer, salivary cancer, colon cancer, rectum cancer, prostate cancer, pancreatic cancer, lung cancer (e. g., non-small cell lung cancer and small cell lung cancer), vaginal cancer, thyroid cancer, melanoma (e. g., unresectable or metastatic melanoma), renal cell carcinoma, sarcoma, glioma, glioblastoma, multiple myeloma or gastrointestinal cancer, especially colon carcinoma or colorectal adenoma, a tumor of the neck and head, endometrial cancer, Cowden syndrome, Lhermitte-Duclos disease, Bannayan-Zonana syndrome, prostate hyperplasia, a neoplasia, especially of epithelial character, preferably mammary carcinoma or squamous cell carcinoma, chronic lymphocytic leukemia, chronic myelogenous leukemia (e. g., Philadelphia chromosome-positive chronic myelogenous leukemia), acute lymphoblastic leukemia (e. g., Philadelphia chromosome-positive acute lymphoblastic leukemia), non-Hodgkin’s lymphoma, plasma cell myeloma, Hodgkin’s lymphoma, a leukemia, and any combination thereof. [0201] There are several anti-CD47 antibodies known in the art. However, the number of available CD47-BDs in the form of antibody fragments, such as scFvs, that can be used as universal building blocks for the construction of multispecific antibodies is limited. It is thus desirable to have suitable anti-CD47 antibody building blocks at hand that can readily be incorporated into any desirable multispecific antibody format, and that are capable to potently block the biological function of CD47, in particular to potently block the interaction between CD47 and SIRPα. Furthermore, said building blocks should have superior 119915P877PC 23.12.2024 Numab Therapeutics AG biophysical properties, such as in particular a high stability, in order to facilitate their efficient incorporation into multispecific antibodies for pharmaceutical development. [0202] In a further aspect, the present invention relates to an antibody binding domain, having specificity for CD47 (CD47-BD), as defined herein. Preferably, said CD47-BD is in a format selected from an Fv, a dsFv, an scFv and a dsscFv, particularly from an Fv and an scFv. Specific non-limiting examples of such anti-CD47 antibody binding domains are the scFvs with SEQ ID NOs: 16, 17, 18 and 19. [0203] In yet another aspect, the present invention relates to an antibody comprising one or two, preferably one, antibody binding domains having specificity for CD47 (CD47-BD), as defined herein. [0204] In another aspect, the present invention relates to an antibody binding domain having specificity for LILRB2 (LILRB2-BD), as defined herein. Preferably, said LIRB2-BD is in a format selected from an Fv, a dsFv, an scFv and a dsscFv, particularly from an Fv and an scFv. [0205] In yet another aspect, the present invention relates to an antibody comprising one, two, three or four, preferably one or two, antibody binding domains having specificity for LILRB2 (LILRB2-BD), as defined herein. Specific non-limiting examples of such anti-LILRB2 antibodies are PRO3603, PRO3635 and PRO4710, whose sequences are listed in table 5.
4 2 , 0 n 2 . i 2 o i t SST SSK SK SE in AV L AV L S T AV L L . 1 3 2 f e ST d . ) I GG SG STG YSG LV YQ I GYQ I GYQ AT R s r D e I GG I GG I GG R C t t b e WI W WI W WI W I YV LK LT a l E LI N E LI N G Y E LI NY KT m i c u l a GY KT GKT CT PPGF N t i S S KS K d GG GG GG GA o t n PAR P A ARA PARA PG g a KG in l d QRCF QCF QCF QYT dr o VY RVY R V VY Q V YSS o b c c n i WV TA W a n MT TA WTA WDT AD MS AD MS S AD VS d w e o YEA YEA YEA WQ SC in h S f NR L SNR L SNR L I e s SYY d e r LS L N S L N SN S s a I D QTA GM I D GM I D GM SF R s D D eu SQ SQ S A VL VL VQ L QE Ce i d AY CV AYV AY P V CQ h s t e ; r S LT C N ST C ST I TTLS e R RK LN RK LN RK VRI S m D L e C hc ( G SSN LSSN LSSN DT L n S GDK GD GT K GDK VF s oi t A G W PQI S G T PI S GPI S SADT g n F QT QT SG inr e e v S S A T A V Y V G L GR V b i n G LF V GRG LF LS GRG SSG 2 7 m t M Y W G GS GS GS PS u n A T I S S S YT GA GA GA SAS n e s YS S I N I S E E S SW SW S QF N S Y S L L S ES ES EWS TR o e r H p L AS TS QS Y Y S G DT VVA V YSVA V YSVA MS YSQP A eh I D G I G Y G R A Q A R G R S Q Q QY TS VQ QY TSQYSVV o t t V Q T V D G g o in tg dro i n ) cc d r K 6 41 a o c de c ) a T _ ta s n n s K 6 S i i g a n ) i g ) a ) g ) g n 4 i r ) ) ) g n 1 1 T 0 1 se mo mo ni r ni r e b g n i r g n i r g n i i r _ r e b S 1 T _ C ds dg d e b e b m l e e s e m m u n b e e m 0 1 m b m b m u n 1 5 n ) e i n b c u n u n o u o T ( G ( o i s d a n H n u n u n 3 _ 3 _ 3 3 G i t at w i n i r e b a u o H o H A; o H o H o H H A; 0 c 0 c 0 c 0 c As u r e f v q ic m tu ( h t o y e g o s d s A; A; 8 3 A ; A ; A ; 8 3 s- o d d l e b e s 1 2 4 RH - 2 6 7 RH - 31H 2 R-814 2 L - 27 2 L - 27 1 L - 3L 4 0 s- 4 s- 4 s- -7 B 0 - B 0 4 0 - B- B-Ce P p i n i e p i t a D7D7D0RD 7 a 4RD8RD8RD0 6 2 6 2 6 2 62 7 r t s 8 e i l i f c mn ax a b – C2 HH ( C5 HH ( C1 HH ( C L2 L ( C L5 L ( C L5 L ( C L1 L ( H- V0 7 H- V0 7 H- V0 7 L- V0 7 Ph e 7 5 1 T e c p E b . 4 D 4 0B 9 a n e s s 9 s 1 le C -t i - 6 1 1 m u u q N e l e S n u b a n 2- T A 0 7 1 2 3 4 5 6 7 8 9 0 1 1 1 4 2 0 2 . 2 ELGE S LLG LE E S L L EG LGTT EGT MD MT EGT MS EGT MS E LG LAD LG LAD LG LAD . 1 Y 3 2 GVYVY YI K YG LAA YVYEA YVYEA YVYEA GTG RVGTSAI KGGE SAVYR GTSR GTSR GTS Y RVREVRVK RTSL GRVNL GVNL GVNR L I K LTI KTI KI T I VKNS I KTLS RKTLS RKTLS LGY TL LGYTY CL LGL LG Y Q L LTL IDN M Y L LGI DN I Y TGM L LGI DN I Y TGM L LGI DN GM KPGKGKQKG KGTGQ L KGSQ L KGSQ L KCGSQ PFPFPGPF PGSVY PFVAY PFVAY PFVALY KAPKAPKFPKA PKFAV PKACV PKACV PACV GGGGGAGGG GAGCT GGS LT GGS LT KGGS LT PGYPGYP PGY P S LNK PGY NK PGY NK PGY N KQTKTKGYKT KGY QSQQSQQTQQS QQTR LS K N QTR LS KTR LS KTR LKS QSSN QSSN QSSN YSDYSDYSSYS YSSSD YSGD QYSGD QYSGD WTWT D WDWT WDGK D WTGPK D WTGPK DTGK SSSSSTSS STGPI ST SSQI S SS I S WSSPI S VQVQVSQVQ VSQF VQVTF VQQ VTF VQQ VTF WC SYWCYWCWCY WQCVR WCYLR WCYLR WCLR ISYS TI SYS TI SYS YI SY S T I SYL YGGS SYG A I STGGS SYGG SYYGG A I STGS I STGS S QAS S S S G S G S GA S GA F A SDQF T A DQAFQF T D QAFG A SWS QFS AS AS DEWS QFDEWS QFEW AESAESADSE SDEA SEVA SEVA SDEVSA QPQPQEAQP AQEVYSAPVYSAPVYSAPVYS CQCQCPCQ CPVYSQCQQYSQCQQYSQC QYS IT LI LI Q I L I QQTVI LQTVI LQTVI Q LQTV TST V TSTTL TS T LQSTTSSSTTSSSKTSSSK IS I S STI S TSSSVTI SGSVTI SGSVTI GSV RT LVRT LVRI SV V TRT L RI S TGAL GSTV GAL GRT LGSTVRT LGAL GSTVS RT LGAL GST DTDTDLDT DLGI G DTGI GGDTGI GGDT I G GFGFGTGF GTGYQGFSYQGFSYQGFGSGYQ VSD ATVSDTVSFDVD SG TST VSFDTS GVD GI G I WSTG GVD GI G I WSTGI GGVD IWSTGI GG IW LSA SG LSA SLGA SG LS A SLGGW AG EI NS LSGW AG EI NS LSGGW AGG EI NS LSGWI N SG SSSG SSSS SGSG SS SSGLY SGGGTSGGLYSGGLYSGGE LY 3 SSSSGTSSSGTSSSCT 7 PA SSPAPSPA PSSK PAGKSPAGKSPAGKS FSSFSASSF SAGGGSSGGGSSGGGSSGGG : QTRQ QS STRSTFQ QS RTRS TFGPRQFGPRQFGPRQFGPR e RGAATRGAATRGAATRGAA c MPMPMSMP MSGQCMSSQCMSSQCMSSQC n e QVQVQPQV QPSRFQPVGRF PVGRF PVGRF u V DG SV DG SV DV GV DG S V DV GGVY G WV AV DG SGVYQ G WV AV DG SGVYQ G WV AV DG SGVY G WV q A e S ) C1 41 ) ) T _ ) G ; 8 G8 ; G8 G 6 8 6 ) p p a 6 6 C a C S _ S _ 1 4 sn S_ S C a p G7 G7 1 T) i a G _ G a p p a 6 A 6 A _ K 6 m 76 7 p K ) ) _ ) _ ) G84 1 o A 6 a _ ( A ( K ( GH HL V ( HL V ( ) H 6 T _ d le s e _3 _ _ 8 G 0 3 c 0 3 0 6V- S r V e - r k3 _ V- 0 e r k3 _K 6 V- 0 e r S _ S 1 b a c n k3 04 1 e G70 1 i r e u As s - c s- c s- _ -G ic 4 7 i n c l- s - i n c l- s - i n c l- s- T _ k S i n 6 l-- A_T _) a v : y no q e s t 0 4 B 0 4 0 4 06 - B- B- BAL4 V 0 ( B L4 V 0B L4 V 0B1 4 0 L V0BLC V1 5 3 0 7 do t i ip d eCu ep 62- 62- 6 - 2 - 6 ( 2 _ - - 3 v6 ( - 2 - v6 ( - 2 - v61 ( - 2- T _ v6 ( 2 _ 3 b 3G _Oi t r c s a P 7 a 7 r L 8 e V0 7L V0 7L V0 7L V0 0F 7c s c s 0 F 7 c s 0 F 7 c s 0 7H F- 0 V c s 0 7 c sHR VP ( n a s e b –Ph 1 D 5 1 T LD 3 b P D 0B-9 a - 9 t i I Q: 0 2-1 1 m u 2 N 1 3 1 4 1 5 1 6 1 7 1 8 1 9 1 n AE SO N7 3 4 2 0 2 SL. SL SL SL G2 GD. 1 GT GDT GDT GDT SG LGG LGG L 2 I A GA GA GA AVSVSAV ILTA ILTVI LT 3 YSG I YG I Y I SG I YG FVKFKGVK GG S Y GY I GG S Y GY ATATGT WD I G E WD WD I G WD R LVY EVY EVY EVY I YGTRGCRGT GG L CG L GG L CG L LGI Y LGI YG KFLKFL LKF KR KR KR R A A A GA A A KA PNPNPN PC G PY PC GPC GPC AKPAPAP I K K QY PY PYY PYY GGTGI GTGI GT IRV QY A V QV QV PKTP SKTP SKTS WT I RAT I RAS I RAS QDQDQD IYD W A I YD W A I YD W A I YD Q A YSYQ FYSYQYSY WA WA A A W WFWF YT DV YT W S DV YT W S DV YT DV AN LSAN LSAN LS S S SS S YQYQYQ NFS L NS L NS L SNS L TCYTCYTCY SFK L FSK FSK FSK I GYI GYI GY S FLS FLS FLS S QTS AQTS AQT GF GF GF GF S A SQ SQ S S FSFSF VKN VN VQN VQN AQDEAQDEAQDE CK K S CK K S CK K S CKS CPCPCP TLSD T LSD T LSD T LS I T D TQ LI TQ STLI TTQ L G G S S S S V VSVS Q Q LTV LTV LTV LV RI SRI SRI S A A EI S EI S EI S TEI S DT LDTDT W W STV STV STV STV GTGLTGLT S S AN PKR PR PR PR VSFVSFVSF A A G KG KG K D D D Y L P V G ATATAT IY YQ Q DT A I G LQ GASV SLQASV SLQASSV LQASSS LGS LGS LG 4 W T Y N TN A L T G Y T PWVG TPWVG TPWVG KPWVSSSSSS 7 KSGSGSG D G G G T S S S S Y I G S S D GSSAV LGSSAV LGSSAV LGSSAV LPSSGPSSGPSSG N S S F G G D S A A SY EYQTEYQTEYQTEYQTQTSFQSFQSF S G G V F I Y I Y Y Q G S I L I L A FA G F Q G N LTGQQ LTGQQ LTGQQ LTG T T QMRMRMR S QVNGGQVNGGQVNGGQVNGGI QSPI QSPI QS G S C R Q R R Q Q S W Q S W Q S W Q S W D V D V DP V ) K6 ) 4 1 ) ) K 6 T _ ) 4 S 1 G )g ) g ) g g n r i r ) ) ) g n 1 T 0 1 8 6 i n in in e g e r n g n g n i r ) _ T _ ) F _ b e r m b e b b m i r i r i r e b C S 1 C C 1 G u e b e b e b m 1 5 0 1 1 5 4 1 7 6 u m n o u m n n o u n o m m m u n G ( T ( G ( T ( A o H u n u n u n o 1 _ 1 _ 1 _ 1 _ ( _ G H H H A A ; o o o H 8 H H H A s A ; A ; 0 1 1 c 0 s - c 0 s c 0 s c 0 0 10 s c s c s c s ic 2 ; A ; t 4 6 6 3 1 A ; A ; A ; 8 3 - - - - - - 1 3 0 3 0 3 0 3 0 3 0 3 0 3 0 u 1 RH - 27 RH - 27 RH - 3H R- 1 2 42 2 72 2 73 L- B- B- B- B- B- B- B-Ce D7D7D7D8 0RDL- 4RDL- 8RDL- 8RD70 0 2 0 2 0 0 0 0 0P p a C2C5C5C1C2C5C5C1H- 7 H- 7 H2- 7 H2- 7 L2- 7L2- 7L2-7r H ( H ( H ( H ( L ( L ( L ( L ( 3 3 3 7 7e h H H H H L L L L V V V V 3 V 3 V 3 V 3 8P5 1 T b 9 a 9 1 1 m u 0 N 2 1 2 2 2 3 2 4 2 5 2 6 2 7 2 8 2 9 2 0 3 1 3 2 3 3 3 4 3 4 2 0 2 G . G 2 S . 1 ALG G GT GT GT GT GT GT GT GT G VSI KSI K GA GA GA GA GA GA GA GA SG L 3 I 2 LTA V GVI LEA VI LEV I YG CG I YG CG I YG CG I YGG I YG SG I YGG I YG SG I YG A G I LTV GKTFKGK I AY Y I AY CY I AY SY I Y SY FK R IYGAT CRGGTG WD I AD D I AD D I AD AD I AD AT GI YQR GI YQ EV LY W EV WV LY E LY W EV LY W EV WV WV LY E LY E LY W EV LY R IYGT LLFL LFL LG GG CG GG CG GG CG GG CG L LG KPAKAKFA KR KR KRA KRA KR KR KRA KR KFA ANPNPN GAC GAC GC GC GAC GAC GC GAC PN KPAKPAKP PAF PAF PF PF PF PF PF PF PP GI G I G I G PTGPTGPT QY RV QY A RV QY A VA QY A VA QY A V QY A V QY A V Y KI G A QVA GPT KTKTKT VAT VAT RVS RS RAT RAT RS RS KT QSDQSDQSD WD WD WD V WD V WD VD VD VD QS QYSYQYSYQYSY I YEA I YEA I YEA I YEA I YE W A I YE W A I YE W A I YEA QDS W W W W W YY WFWFWF YR L YR L R L R L R L WR L WR L WR L WF ANANAN DS DS YDS YDS YDS YDS YDS YS AN LSQLSQLSQ SN SN SN SN SN SN SN DSN LS YT IGCY YT IGCY YT IGC NM NM NM NM NM NM NM NM Y Y FQ TQ L FQ FQ FQ FQ FQ FQ FQ I GCY SYSYSY SF SFL SFL SFL SFL SFL SL SL SY QTQTQT GYV GYV YV YV YV YV FYV FYV QT SASASA ST ST GST GST GST GST GST GT SA AF QD CEAF QDEAF QDE AANK AN AN AN AN AN AN SAN AFD S AKS AKS AKS AKS AK AK AK QE ITP TQC ITP QC ITP C Q S LN C D S LN C D S LN C D S LN C SLN CS SLN CS SLN CS SLN C ITP Q VLSTVLSTVLS R LR IS R LR IS R LR IS RD LR D IS R LR D IS R LR D I R LR D I R LR TL I VS RDI S GTRI LDS TRI LDS T L SGT S F GT S F GT S F GT S F GT SS SS SS RI S F GTF GT GT DT L VTGVTGVT GR GR GR GR GR GR GFR GFR GT SFSFSF PG PG PGSPGSPG PG PGSPGSVSF AD SLTAD GSTADT Q VQSQ ASVQSQ ASVQ Q ASVVQ Q ASVQSQ ASVQSQ ASVQ Q ASVQASADT SSLGSS LGS L V GWTL V GWTL GWKL V GWKL V GWTL V GWTL V GWKL V GWKS LGS 5 S 7 PGS SSS QGPGS SSSG GPSS GSAV LGSAV LGSAV LGSAV LGSAV LGSAV LGSAV LGSAV LSAG G GSYTG S QGSYTG QGSYTG QGSYTG QGSYTG QGSYTG QGSYTG QGSYTP QGSG TSFQTSFQTSF E VLTQE GV LTQE GVTQE GVTQE GVTQE GVTQE GVTQE GVTQQS GTF MRSMRSMRS QNGWQN LN LN LN LN LN LN MR IQ I Q I Q GWQGWQGWQGWQGWQGWQGWI QS DP V DP V DP V V ES SL DV ES SL DV ES SL DV ES SL DV ES SL DV ES SL DV ES SL DV ES SL D DP V ) ) ) K C 6 1 K 6 4 4 1 1 4 1 ) T T _ _ ) ) T _ K 6 S G8 ) p a K 6 S 1 4 1 1 6 0 p 1 F a C 4 _ C a 1 p T 0 1 ) T _ T _ p ) _ T _ C S 1 C G a 7 p a C S 1 C 1 6 p 5 0 1 1 5 A a K ( K ( _ 1 5 0 G G ( 1 1 T 5 ( G ( G ( T ( G ( _1 _ 1 8 6 9 _ 9 _ 9 _ 9 1 . 9 _ 1 . 9 _ 1 . 9 _ 1 . 1 . G 0 0 F 0 0 0 0 0 0 0 9 0 99 c c _ c c c c c c c c 0 c 0 c As s- i c 3 t 0 s- 3 -G B 0 3 0 7 6 s- 30 s- 30 s- 30 s- 30 s- 30 s- 30 s- 30 s- 30 s- 30 s- 30 - B- B-A( B- B- B- B- B- B- B- B BB-Cu e 02 02 0 2_ 1 0 2 0 2 0 2 0 2 0 2 0 2 0 -0 -00P p 7 a r L- 73L- 73L- 70 3 c sH- 73 H- 73 H- 73 H- 73 H- 73 H- 7 H2- 7 H2- 7 L2- 72- 77 8 e h V V V V V V V V V 3 V 3 V 3 V 33P5 1 T b 9 a 9 1 1 m u 5 N 3 6 3 7 3 8 3 9 3 0 4 1 4 2 4 3 4 4 4 5 4 6 4 4 2 0 2 . GG 2 SL G V SG L GG V SL G G S V SI K SI K GD S T GD S T GD S T GDT GSG LGG VSLV . 1 A 3 2 I LT A FV I LT A V I LT A V I LE A V I LE G V I YA G G I YA GA GA A G I YG I YG I LTA ILTV AKT GK GK FKT GKT SG SG SG SG FVKFK R IYG GT GT C RG G AG GG I AY I AY I AY I AY ATATG LLG I YT R G I YC R G I YQ R G I YQ WD G EV WD Y EV WD EV WD Y EV RG Y I YTR GI YCG KF L PA LKF L A LKF L A LKF L A LKF L LY L A GG G LG L LFL LF KR CG KR GKR CKR KAKA PN PPN PPN PPN PPN GA GA GA GA PPNPN KP P P P P I K K K K PCF PCF PCF PCF KPPKP GG GI G GI G GI G GI G AY AY AY AY GI GGI G PTT PTT PTT PTT PTT QV QV QV QV PTTPT KQS KS KS KS KS RVA R R T VA VA RVA KSKTS QD QQD QQD QD QD T S S WD WD WD WD QDQD YSY YSY YSY QYSY QYSY I YEA I YEA I YEA I YE Q A YSYQYSY WF WF WF WF WF WFWF AN AN AN AN AN WYR L WYR L WR L WR L ANAN LSQ LSQ LSQ LSQ LS DS DS YDS YDS LSLS YTC YTC YTC YT YTQ SN SN SN SN YTQYTQ IGY I GY I Y I CY I CY NM NM NM NM I CYI CY SY SY GSY GSY GSY FSQ FQ FQ FQ GSYGSY QT QT QT QT QT FL S Y FL S Y FL S Y FL QTQT SAF SAF SAF SAF SAF GV GV GV GYV SASA AQD AD AD AD AD SAT SAT SAT SAT AFDAFD CE QE QE QE QE N N N N ITP C I P C AK AK AK AK QEQE I P C I P C I P CS CS CS CS C I PC I P TQ VL TQ S TVL TQ S TVL TQ S TVL TQ S TVLS S LND S LND S LND S LN T D TQ VL TQ STLS RDI S T RDI S T RDI S T RDI S RDI S R LK R SI S LK R SI S LK R SI S LK SI S RDI SVRDI S GLT GL GL GT L GT L GT GT GT GT GT LGT L VSF VTF VTF VTF VTF GF GF GF GF VTFVTF AD SAD SD SD SD PR G PR G PR R G PG SDSD SLTG S LT A G S LT A G S LT A G S LTG Q VQSQ ASVQSQ ASVQS ASQ VQSSA SLTA GS LTG SS A SS SS SS SS L VTL VTL VKLAVSSSS 6 7 PG SS APGS APGS APGS APG G S GWS GW AV LGS GW AV LGS GW AV LGSK VLSPGSSPGS QG TS S F QGS SQGS SQGS SQGS GS R TFR TFR TFR TFR EYTG QGS TQEYTG QGS TQEYTGA QGS TQEYTSGSG QGQSQS TQTFTF MS MS MS M M V LNGV LNGV LNGV LNGMRMR IQP I QP I QP I QSP I QSP QG Q Q Q Q S LI QS D VI QS D V D V D V D V D V Q SW L QG SW L QG SW L GW P DP V ) C1) ) 41 C 1 K 6 ) T G _ 4 1 ) ) 4 1 T 8 ) GT _ 6G 8 6G ) p p a ) a) C p a K 6 _ 41 S 1 ) C1) F _ 8 6 F F _ 8 6 Cp a a a p C 0 p a p T _ 1 T ) 41 C 1 G7 _ GF G 7 _ pC pa p a p a ) C S 1 _ C C 1 T ( 4 _ 1 6 1 . 9 T ( A ( 7 _ 9 _ 6 6G7 1A A ( .9 ( _ _ 6 a p 1A K ( .9 ( _ _ a K _ 1 . 9 K ( G K 8 _ 1 5 0 1 1 5 4 1 _ 6 G 9 G F 8 6 ( 8 _ 8 T ( _ G ( _ T ( _ G 00 090 090 _ F 8 8 8 8 cc cc 00 G _ 1 1 1 1 1 1 As s- i c 3s- 3 s- 3s - c 3 s - c 3s - c 3 s - c 3s- 3 -37 6-3G c 7 s - c s- c s- c s- c s- c s- t 0B0 00 00 00 0A -B- B- B- B- B- BB B( _0B6A 3 0 3 0 3 0 3 0 3 0 3 0 ( B B B B B- B-Cu e P p 0 a L2 - 0 72 - 0 7L2-0 72 - 0 7L2-0 - 2 - 0- L2 - 0 - - 2 0 L2 . - 1- 9 0 0 2 _ - - 9 0 - 0H2 - 0 - H2 - 0 - H2 - 0 H2- 0 L2- 0 L2-7r 3 3 33 7 3 7 3 7 3 7 3 7 3 c s 7 3 c s 7 7 7 7 7 7 7 8 e h V V V V V V 3 V 3 V 3 V 3 V 3 V 3 P5 1 T b 9 a 9 1 1 m u 7 N 4 8 4 9 4 0 5 1 5 2 5 3 5 4 5 5 5 6 5 7 5 4 2 0 2 G. 2 SG LGSG LG G G VSI KSI K SG LYT G DV . 1 A I S SG LYT G DVS SG LYT DV G S SG LYDLS G N SG LYDLSN LVTA VI LTA VI LEA VI LEV AV ILTS AV NS L I LTS AV NS L I LTSNS L AV ILTSNM A ILVTSNM 3 2 GGK RTGGKTFK ATGK GT FV AKF TSK L GV GKF TSK L GV GKF TSK L FV AKF TSFQ L GVKFSQ Y GTFLY IYGTR L I LGYGCR L I LGYGQR LGI YGQ R LG I YGF TGS LGSF R VQ I YGF TGS N LGSF R VQ I YGF TGS N LGSF R VQ I YGTGSVT R IYGTGSVT N LGAN LGAN KFKFLKFLKF LKFKK LKFKK LKFKK LKFAK LFAK PANPANPANPAN PANCS PACS PACS PACS KPACS PPPPPPPP APTS ANPTS ANPTS PNPS LN PNPS LN KGI GK TGI GK TGI GK TGI G K T GI G LD K TS LV GI G LD K TS LV GI G LD K TS LV GI GTR LDR KGI GR LDR PKTP QSKTP DQSKTP DQSKT P DQS KTT QSEI S ST PTTI S PTTI S PTSI S PTTSI S V KQSESTV KQSESTV KQSGT GF KQSGT GF Q D DPR DPR DPR DPR DPR YSYQYSQYSQYS QYSKG QYSKG QYSKG QSQG QSQG WF Y ANWF Y NWF Y NWF Y N WFNV LQ Y AS FVQ YFVQ Y S YF LSALSALSALS ALSG SWANL SGASSWANL SGASW VANV LQASYY SWF SGWVANV LQASS SGWV YQYQYQYQ YQPWVTLQPWVLQPWKLQGSTLQGST T GCTCTCTC TCGS YTCGSTYT GS YT GAV LYT GAV L I Y Y Y Y YSAYV L YSAYV L CYSAYV L CYSYT CYSYT SYI G TSYI G TSYI G TSY I G E T SYTQQTI G E GSYTQQTI G E TI G GSYTQQGSYTEQTGI GSYTEQTG QAQAQAQA QALTQQALT QALTQQAV LNQQAV LNQ SFSFSFSF SFQNGSFQNQGSFQNGSFQGGSFQGG ADADADAD ADVGS ADVGS ADVGS ADVSWADVSW QEQEQEQE QEQSWQEQSWQEQSWLQEESLQEESL C ITP QC ITP QC ITP QC ITP Q C ITPS L QGGDC ITPS L QGGDC ITPS QGGDC ITPS QGGDTC ITPSGGDT TLTLTLTL TLGGTTLGGTTLGGTTLGGA Q LGGA VSVSVSVS VSGI YAVSGI YAVS I YA S I YGTS I YG RDI SRDI SRDI SRDI S RDI SGSSGGRDI SGSSGGRDI SGGSSGV GRDI SGGC IAGV YRI SGGC IAGY GT L T L T L T L T L I GY T L I GY T L I GY T LS DDT LS D VTGVTGTGT GTGWDGTGWDGTGWDGTGWVGTGWV SFSFVSFVSF VFG VVFG VVFG VVFGE LYVFGE LY ADADADAD SDGE LYSDGE LYSDGE LYSDG GSDG G SLTGS LTGS LTGS LT A G S LTGGSGKGA SLTGGSGKGA SLTGGSGKGAT RS LGGG SKRA SLTGGG SKR SS SGSS SGSS SGSS SG SS SGG R GGACSS SGG R GGASS SGGGGACSS AGGGA GPCFSS AGGGA 7 GPCF 7 PSSGPSSGPSSGPSSG PSSGGPPYPS PPCYPS PPYPS AYPS AY GQYSGGGQYSGGQYSGGQVSGGQV QTSFQTSFQTSFQTSF QTSFSGI RVAQTSFS VQSG GI RATFS VQSG GI RATFSRAQSG GVTTFSRVAT MRMRMRMR MR Q GWTMRGWTMRGWSMRGWDMRGWD I S DP VI QS DP VI QS DP VI QS DP V I QS I YD DP VG G WA AI QS I YD DP VG G WA AI QS I YD DP VG G WA AI QS I YE DP VG G WA RI QSGI YE DP VG G WA R ) C1 41 ) ; ) ) T G _ ) G p 8 G ) a 8 G 6 8 G 6 8 6 6 8 6 p a C F _ F G _ F _ F_ F G _ C a a p p 7 G G 6 7 6 7 6 7 G 6 7 p p a ) ) A_ ) A_ ) A_ A 6 a K _ _ GH HL HL) H ) HL ( 8 A ( _8 K ( _8 8 6V-r VV ( VV ( K V VV ( e1 -r e_ 1 -r e_ 16 41 -r e9 -r e_ 9 G 1 c 1 c 1 c F _k i s n0 c k i n0 c k i 0 c T _ k i 0 c k i 0 c A s- s- s- -G ic 3 7l- s- l- s- nl- s- nl- s - ) nl- s- t 0 3 B 0 3 0 3 06 - B- B- B-AL3 0 ) 7 L3 0 L3 0 S 1 L3 0 7 4 L3 0 ( V ( B- 9 9 V ( B- V ( B- 0 1 V ( B3 1 V ( BCu e 02 02 02 0 2_ 8v0 2O v0 2 v0 - 2T _ v0 - 2O v0 P p a L- 7L- 7L- 7L- 71Fc- 7R Fc- 7 F-H F-R F2-7 7 r 8 e V 3 V 3 V 3 V 3 c s s 3 P ( s 3 c s 7 3 V c s 7 3 P ( c s 7 3 Ph 5 1 T b 9 a 9 1 1 m u 8 N 5 9 5 0 6 1 6 2 6 3 6 4 6 5 6 6 6 4 2 0 2 G 2 G LYLS GYL GG LYL GGYL GGYL GGYL GGYL. S . 1 A 3 I 2 LVD TSN GSLDSS S DSSN SLDSSN SLDS SLDS SLDS GVNFM AV ILTVNN AV FM I LTVNFM A ILVTVN AV FM I LTSN AV VNM I LTSN AV VNM I LTSN VNM GKTSFQ L FKSFQ L GGKTSFQ L GGKTSFQ L FAKFSQ L GKFSQ L GKFSQ L R Y AT Y Y Y TFY GTFY GTFY IYGTG LLGSV RG FAT I YTGSV RG AN LGAT I YTGV RG AN L LGS FAT I YTGV RG AN L LGS FAT I YTGV RG AN L LGST I YTGSV RG T I YTGSVT FAN L LGAN L LGAN KACKS LKFACK KACKS KACKS KAAK KFAAK KFAAK PPNSN PNSSN PNSN PNSN PNC KPLD PPLD PKPLD PKPLD PKPSS LN PNC D PKPSS LN PNCS D P N KPS LD GI G PTR LR K S KTGI S GI G T PTR L TSR GI S GI GTR LR T P S KTGI S GI G T PTR LR S KTGI S GI G T PTR LK I KTSI S GGTR LK IS GI GTR LK IS GT PKTSGT PKTST QSDGF KQSDGF QSDGF QSDGF QSDGF QSGF QSGGF QSPRG QSPR QSPRG QSPRG QSPR D G QSPR QDSPR YYQ WFV LQSYYQG YYQ AS FV LQSWFV LQSYYQ SWFV LQSY S YQ WFVQSYYQG SWFVQSYYQGS SWFVQS AN LSG VWNGASVANGAVANGAVANLAVANLAVANLAV YTQGWSKA CGAV LS LYQGWSTLS WTLS WKLSGWTLSGWTLSGWK TTCGAV LYTQGS CGAV LYTQGS CGAV LYQGS TTCGAV LYTQCGS GAV LYTQCGS GAV L I GYSY YSYTI GYSYTI GYSY I YSYTI YSYTI YSYT SY QTE VQGI G SALTNQSYTE GQAV LQTG YEQG YEQGGY NQST GQAV LTNQST GQAV LTNQSTE GQAVQ LTGG NQSYTE GQAVQ LTGG NQSYTE GQAVQ LTG NQG AFQGWSFQG SFQGWSFQGWSFQGWSFQGWSFQGW QD CEVESSLADVS DQEESW LAQDEVESSLADVS DQEESLAD DQEQ-SSLAD DQEQ-SSLAD S DQEQ-SLD ITPS QGGGTC ITPS QGGDCPS GTI TQGGGTC ITPS QGGGTC ITPS QGGGTC ITPS QGGGTC ITPS QGGT TVLSGGI YAGTVLSG A GI YGTVLSG A GI YGTVLSG A GI YGTVLSG A GI YGTVLSG A GI YGTLSGG IYAG RI S CGRI S SGRI S SGRI S SG I S SG I S SGVI SGSG D GSI AYD GSI AY GI AY GI AYR GI AYR GI AYR GI AY GT LG DGT L DDT LS DDT LS DDT LS DDT LS DDT LS D VT WVVTGWVGVTGWVGTGWVGTGWVGTGWVGTGWV SFGE ADGLY FG GS E ADGLY FG GS E ADGLYVFG GSDGE LYVFG GSDGE LYVFG GSDGE LYVFG GSDGE LYG SLTGGG SKR T SS AS LGGG SKRS LTGGG SKRA SLTGGG SKRAT AS LGGG SKRA SLTGGG SKRA STGGG SKRA AGGG GPCFSS AGGGA GPCFSS AGGGA S GPCFSGGGPC S FSGGGA PC S FSGGGAL PC S FSGGGPC 8 F 7 PSS A GGQYVPSS A GGQYVPSS A GGQYA VPSSGA GGQYS VPSSGA GGQYS VPSSGGAQYS VPSSGGAQYV QSGRAQSGRAQSGRAQSGRAQSGRA GSGRA GSGRA TFSVSTFSVTTFSVTTFSVSTFSVTQFSVTQFSVS MRG IQSGWD IYEMRG SGWD IYEMRG SGWD IYEMRG SGWD IYEMRG SGWDT I EMRG SGWDT I EMRG SGWD I E DP VG G WA RI Q DP VG G WA RI Q DP VG G WA RI Q DP VG G WA RI Q DP VGY G WA RI Q DP VGY G WA RI Q DP VGY G WA R ; ) G ; G8 6 8 G ) ; 6 8 G G F_ F 6 _ F G _ 8 6 8 F 6 G _ F _ 76 7 G 6 A 7 6 G G A 7 6 7 6 ) A H_ L ) ) _ L ) _ L ) ) A_ ) A_ V-r V ( ) _9 K 6 HV 4 - r 1 HV ( .9 V-r _ 1 HV ( .9 V-r _ 1) .9 K 6 HV HL VV ( HL VV ( ) K 4 - r 8 -r _ 8 -r _ 86 4 G e k 0 c1 T e k 0 c e k 0 c e k 0 c1 T e k 1 c e k 1 c e k 1 c1 A i s ns - _ i ns - ) i ns - i ns - _ i ns - i ns - i ns- T _) i l- c L3 t u V 0 S 1 l- 3 0 0 3 l- 3 0 l- 3S 1 l- 3 l- 3 l- 3S 1 5 4 ( B v-0 0 1 L T V ( B v-0 2 2 L O V ( B L v-0 V 0 ( B v-0 0 1 L V0 ( B L v-0 V 0 ( B L v-0 V 0 ( B v-0 0 1 2 4Ce P p 7 a F2 - _ H F2-R F2- F2- T _ H F2- F2- F2- T _O HR 7 r c s 7 3 c s 7 3 8 e h V P ( c s 7 3 c s 7 3 V c s 7 3 c s 7 3 c s 7 3 VP (P5 1 T b 9 a 9 1 7 1 m u N 6 8 6 9 6 0 7 1 7 2 7 3 7 4 2 0 2 . 2 AL. 1 VN AL L L N AN AN S SA VFA VVFA VVFA VVFA AL L 3 2 C IADS C WY I ADS C Y I ADS C IAD TG S SL T AVT GG L GVT ELA W D A WYA WYA KVK KVK K E LD E LDK E LDK I YT I YT GKR CK KR GR CR L LGT L LGT GA PC GA K PC GA K PC GA K Y PC PAG K F PAGF PY PY P Y KV KV QY RV QYV QY P V QYV GN VA RVA RVA RVA PKI D GN P D KI DD WT CDA WT CDA WS CDA WS CD QT A QI I QQTI I MA A A A YY YY DT M V DT M V DT M V DTV WGQ WG YS YS YS Y A AQ G G G GS LQC LQ SSS L SS L SS L SS L YDY YDCY FK L SFK L SFK L SFK L I Y I Y SS SS SS NT NT F F F SFS SA SA GVQ GVQ GVQ GVQ QSFD QSF SN SN SN SN A D AK AK AK A E AE K K K KK QP QP CS TS LD CS S S C K TS LD C TS LD C TS LD I TQ L C TS I TQ L TS G S K LTI ST S LK IS S LK IS S LK IS VRI S T VRI S T A EV TTV TTV TTV DLT DLT W SR ESR ESR ESR GVF GVF S P A KG P C Y V KG PKG PKG SDT SDT LKA V LKA V LKA VKA AG AG M Y LN A D L V S S S N GWSGWSGWSL S GWSS LS S SG LS SG 9 7 Y I TD FA Y G V D D I D PSV GATPSV GATPSV GAKPSV GAKSSG SSG S S S I N S S D SY S A A TII EYV LSY F I TEYV LSY I TEYV LSY TEYV LP TSS P QF SS QF S G Y L L QTGQT Q I T Q I T TR TR S A A Q F V D S T T Y G G L QDVQL QDG VQL QDG VQL QDG VQMSP MSP G V C K R A S Q A K G K Q Q V QSG G WV QSG G WV QSG G WV QSG G WI Q DV G I Q DV G ) K6 ) 4 1 ) ) K 6 T _ S1 ) G )g ) g g 4 1 0 1 8 6 ni g n r n i i r ) r e b g ) ) n n i i g n i g n i r e ) T _ T C S _C S _ se e b e m b m r m u n e r b e r e b m 1 1 0 1 G7 m b m b 5 m u n G 1 T 5 6 cn u n u n o u u u o ( _ ( _ G ( _ A ( eu o o H n o n o n o H 8 1 8 8 8 8 _ 8 GA q e HA HA A; 8 H H H A; c si s ; s - ) 1 c s- 1 c s- 1 c s- 1 c s- 1 c s- c 2 ; 3 1 A ; A ; A ; 8 3 2 0 2 t 9 2 0 2 ) ) ) 0 1 9 2 0 2 0 2 9 2 0 1 9Cu d e 142 6 73H- 1 2 42 2 72 2 73 1 L G3 e s a RH- RH-R8RL-RL-RL-R-7 -31 G G8 O -3 -33 G G3 O -3 -31 G8 O -33 O P p 7 a 7 r b D7 – C2D7 8 e 2 HH 5D0 1D ( C 4 HH ( C HH ( C L2D ( 8 LC L5D ( 8 LC L5D ( 0 0 LC L1 L ( H- V3 3R ( 0 PH- V3 3 H0- V3 3R ( 0 PH- V3 3 L0- V3 3R ( 0 PL- V3 3R P (Ph 0 5 1 T b G-9 a 9 3 1 1 m 0 u -3 N 3 4 7 5 7 6 7 7 7 8 7 9 7 0 8 1 8 2 8 3 8 4 8 5 8 6 8 4 2 0 2 . S 2 A S S . 1 L A A SGMA L AGDT SAGGMDAT SAGGMDAT GL L 3 T 2 GL GYV LGYV L YV GVT TSAI KT EGI K TLGS TLGS TG LGS E SVTSS L GVTSS L GVTSS L KV IYKKVG L I LTYKKV A TI YK K T GL I VS LGLG YKFSK L GKVSFK GKVSFK LT S I YKSLS I YKSLS GFV LTGFV LTGFV KCKQLKQ LKTGSQ L G N KTSQ LKTGSQ PAGPGPG PG G N G N KFVAKFVAKFV AKFAK P VK FAK P AK CS AKVKS AKFVKS GN P GNGN GNT KI DPKI DPKI D PKI D LSD GNC T SK PI D LSD GNC K PI DT LSDK QDTQDQD QDLI S KQDS LI S KQDS LI S QI I QTI I QTI I TI I TT TI I TT TI I TT YYYYYY QYYESV Q E R YYSV Q E R YYSV WG A WGWG GP GP GPR Q Q Q WQKG WQKG W G LQALQALQ ALQVK ALQVK AQQK VK YDCYYCYC YCL GASSYCLASL SYCLASS INYDY TI NYDY TI NY DYPWVDYGPWVDYGPWV T I NYTGSTI NY STI NY SK S QA SFS QAFS QAF S QAFSA EYYV LSTAGSA TQFEYYV LSTAGSAV TQFEYYLT ADSDSD SDQ QEPAEPAEP AEPLI TDGSD QAEPQ LI TDGSD QAEPQ LI TG DQ CQQCQQCQ QC QVVGQC QVVGQC QVVG IT LI T LI L I Q LQSWI Q LQSWI Q LQSW TSTSTTS TTSSGLTTSSGLTTSSGL L VI SVI SVI S VI SGANVI S AN I S AN G N RTRTRT RTGVFRTGGVFVRTGGVF N F DL GTDLDL DLGVCADLGVCADLGVA A GS VFGT SDVFGT DVF GT D VFDG D SI ASGT VFDG D SI ASGT VFGCD W T SI AS S E ATS SLGATS SS LGAT S SSG AT S SGGWYST SGGE LA GWYSDTGWY DAG KSSGGE LADAG KSSGGE LA A P D Y D K C YT G SG SSSGL SSSG LG G SS SSSGSKRLG G ASSSGKRLG G ASSGKRA MY N G A L L 0 8 PSGSPGPG PG GCPGSGCSPGSGC Y G T NS QFSS TRQFSS TRQF SSGP TR QFGPY TRGQYSSGP VQFGPY TRGQYSSGPY VQFGP RGQY N W W V N A T I G S T G Y S EL MSMSMS MSGRAMSGRATSGRA F V D Q T IQP DV GI QP DV GI QP DV G I QP DVSV GG GWT CD AI QP DVSV GG GWTM CD AI QP DVSV GG GWS S CD A F L G M R C A S R A S Q A K ) C1 41 ) ; T_ ) G8 G G8 6 ) p p a 6 8 S 6 S sn ) S a C _ _ C a p G _ 7 G i a m ) g ) g g n i r ) G a 7 p p a 6 A 7 6 A o 6 p d in r in e g e r b n i r A a ( K _ 8 K ( _) ) _ _ GH L ) _ L V HVV ( HVV ( ) l e K b a s e b e b mu c mu mu n e b o m G 1 8 8 6-r 8 -r _ 8 -r _ 86 i r n n n c 1 c Se k 1 c e k 1 c e k 14 1 a v e u o o H u A n o As s- i c 2 0 s - _ 2 -Gi 0 2 0 7nl- s - ) i nl- s - ) i n c l- s- T _) yd q e H H ; s A ; A ; 8 H 3 A ; t G6AL2 0 2 G9 3 L2 0 4 G2 7 L2 0 S G1 1 0 9 o 8 b d e 1 2 42 6 731 H1 2 42Cu G- e 3 G-3 -3 ( _ V ( v-31 O V ( v-33 O V ( v-31 T3 i t n s a RH-RH-R-8RL-RP p 7 a L0-L0-L0- 8 1F0-R F0-R F0 - _O a HR 2 b DC7 2DC7 5DC0 1DC4 2DC7 r 8 e Ph V3 3 V3 3 V3 3 c s c s 3 3 P ( c s 3 3 P ( c s 3 3 VP ( BR – 1 HH ( HH ( HH ( LL ( L5 1 T b I L 1 L 9 a - E-8 9 m 7 8 9 t in 1-1 1u N 8 8 8 0 9 1 9 2 9 A 8 6 3 9 4 9 5 9 6 9 7 9 4 2 0 2 G. 2 VG G L VGL GVG SV L ET SV V ELT S V ET SIE SIE SV LV EV EV ETVNQ L . 1 L3 MT LT LT L L L L T TK TK TK TK TK KFY 2 CW M IAT CW MCW ST GT GT ST GT TSTSV Y I TY TY AG GG GG AG GG AGFT WD AD I AD KTG KTG KCG KQG KQ KTGSNK E W LRA E LR W A E I LRA Y LFV I Y LFV I Y LFV I Y LFV I YG LFV I YG LFVAASNS GR GR CR LKY LKY LKY LKY LY LYCDS KA GC KA KA PGY PG PG PG KPG KPGS LKVT PF GC AY PF GC V AY PF P V AY KN PY V GPG KN PY S GPG KN PY S GPG KN PY PY I S S GPG KN S GG KN S GGR LT SSFV L RT QA QA QA R R R P P G G RVT R K K K KR KR KR GQ D VS R D VSD QS QS QS QS QS QSGPNG WCEA WCEA WCE QS A YG QSG QSG QSG QSG QSQAW MYR L MR L MR L WA YA YA YA YA YGAV YS YS YS AT W LS AT WT WT WT WTLW LS ALS ALS ALS ALSGSL GAN YFG NN YNN YNN NQC NQC NQC NQ NQ NQGYS NM NM NM SY SY SY SCY SCY SCYSTNT TQ TQ TQ I GY I GY I GY I GY I GY I GYE E FSL FY FL GV S T FY FL V SFY ST ST ST ST ST STV V QAF QAF QA QA QA QALGPD QWG SAN GT SN GT S SN AD SAD SF AD SF AD SF AD SF ADQAGG QE QE QE QE E ESVL AKS AK C SNSAS AK NSASNSCP I Q L CP I Q L CP I Q L CP Q I Q L CP Q Q CP QGGLT MW LDKSC VS LDKSC VS LDKST VT T VS TS TTS TTS I TTLS I TTLSGGCT IAY RLI STR LI SKR LI SKRI S V TL RI S V TL RI S V TL RI S V T RI S V T RI S TS D GWR STFV LSTV LSTV LD D D DL DL DLGEA V G TGFTGFTGTF GTF GTF GT GT GT LR Y GRGGRGGRGVD VD VD VFD VF VFGGA G PGQPGQPGQST ST ST ST SDT SDGKC Y QNGQNGQNGAG AG AG AG AG ATSGF N A A A S LS S LS S LS S LS S LS S LGSGPY G V S LWW GSLV LWW GSLV LWW GSLSSG S S S S S GAV S SGS SGS SG SG SGGQA 1 8 R GANFGANFGANFP P P PS PS PSGRT SE S GYGGYGGY SG SG SG SG SG SGSVDE L S Y Y YGQS G SETSTSETSTSETSTTF QS TF QS TF QS TF QS TF QS TFGGWCA T N G AT V LGE N PV LGE N PV LGEPMRSP MRSP MRSP MRSP MRSP MRS R PGMLS G S QWDQ DQ DQ DV Q G DV Q G DVG GLQ V DV Q G K DV Q G K DVGYN K Q Q A G QW A G QW A GV G LV G LV V V V GG LY N M ) K6 ) 41 C 1 ) ) K 6 T _ 4 1 ) 4 1 S 1 ) T _ ) p 0 G8 G8 ) p a C g ) g n n i r g n i r T _ 1 T 6 6 _ S a _ S _ C a p p e b i r e e b S m 1 0 C1 5 G a 7 G7 p p a K) m b m u n 1 6 6 a _ GH un o T ( G ( A ( A ( K ( V o u n o H 7 _ _ _ _ _ 8 0 7 7 7 7 7 7 6- S r e7 G H H A; c 0 s s c 0 s c 0 c 0 c 0 c 0 c _k i n0 c A A ic ; A ; 8 3 - - s- s- s- s- s- -G7l- s- tu 2 7 2 2 73 1 L 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 16L1 1 P p L- - E- E- E- E- E- E E EA( V ( E7 a 8 L- 8D7 0 8 1- 8 - - - 1 _ - -Ce RD R 8 1- 8 1- 8 1- 8 1- 8 1- 8 1- 7 0vF8 1-7 r 5 8 e L ( C L5 L ( C L1 L ( H V8 6 H V8 6 H V8 6 L V8 6 L V8 6 L V8 6 L V8 6 L V8 6 c s c s 8 6 Ph 5 1 T b 9 a 9 m 0 1 2 3 4 5 6 7 8 1 1u N 8 9 9 9 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 4 2 0 2 S. 2 EV LTN TVKTFQ S L EV LTVN S TQ L ETN LVT DTF KFQ L I DY . 1 GTSYV TKTFSYV TTSFY I Y I T 3 2 GG KTFT GGGFT GGGGVT C IAA G IYGGSNK KTGGSNK KCGSANK WT LLFVAS Y SI Y LFASSI Y LFASS EV KYACNDSLVYACNSLVYCNS LG PGSKVK D PGSKVKPGS LDKV GKY PYLI STPYLI SK VPYR LI SK V GG KN G R LTV LKNR LTLTKNSTL R T PA PGSSFRTGGSSFRGGGSGFG AC KRG QSGGGQPKRGGQPKRGR QF PGQ RY QSPN YGQAGQSG WQSPNGQSSQNG VV YGQAWQ A LYGV L WL WAT WAV LWSLNWAV LWSNWAGWSN CD AT LSGAFAT LSGAFATSGAF MYEA NQGY SCGYGSNQGYYGSLNQGYGS D IGYSTTSCGSTTSCSY ETT ASR LS SYTENE QAV I LGPGYYENEI GYY DSTV LGPDSTV LNE LN QGPD DFM SFQWAGQAFQWAGQAFQWG GQ ADQGGSADQGGS L LADSAGGL SY QEPSGVLTQEPSVTQEPGVT AAV C L T C GLWC GLW CT ITQ LGGMW CTI TTQ LGGMCTI TTQ LGGM N CT S LK VSGI AY S DV GI AYDVSSI AYD RS RI S DT GLSGWRRI S DT LSGWRRI SG ADT LGWR LN SD VTFGEA TGE EA G RK SDGLRG GGKAVF SDGLRG GGKAVTFGLR K G SDG I S SCKA A L PT ATSGCFATSGCFATGGCF W TF QF SLGGPYS LGGPYS LGGPY T Y V LRS SSGAV SGGQASSGAV SGGQASSGAV A SGGQA C Y I T A G A L GKS VL 2 8 PSGRTDPSGRSPSSRS M Y G Y T GA SGSV QSGWESGSVD QSGWESG QSGVD Y T WE D N T T Y I G S S N GSWTV LT TFGCATFG ATFG A A D V S A A S EAG MR QSGMR L R CR SMSGML RGCR SMSGML S D L L S L I D G VP Y Q T T SD V LYYQG DVGY GG LYNQ N MVP DVGY GG LYNQ N MVP DVG GLY VYN D N M F G I Y C G S R A S Q A G R G Y R S Q Q ET QN DW L ) G ; 8 G 6 8 S 6 ; _ S G _ C 1 4 ) ) 7 G 1) g n g 6 7 T _ K ) g ) g i r ) g ) g ) g n i r ) A 6 _ A 6 in r in r e b n i r n i r n i r e b L ) _ L ) G84 1 e b e b m e b e b e b m HV-r V ( HV ( ) H 6 ST _ s e m m u n m m m u n e_ 7 V-r _ 7 K 6 V 41 -r _ GS 1 c 0 n u e n u n o u o o H u n o u n o u n o o H 5 G k A i s n0 c e s k - i n0 c e s k - T _ i n 7 -61 AT ) q e H A H A A ; 8 H H H A 0 ; c s- i l- c L1 t 1 l-L1 1 S 1 l-L1 1 _ L_ C4 4 s u V ( E d ; 2 ; - V ( E- 0 1 V ( E-V( 1 5 7 3 e s 14 6 H 27 3 1 A ; A ; A ; 8 31 4 0 H3H- 1 2 42 2 72 2 73 L- G-Ce v8 1 v8 1T _ v8_ 7G _Oa RD- 7RD- 7RD8 0RDL-RDL-RDL-RD7 6 P p 7 a r Fc- 8 Fc- 8H Fc1- 80 cHR b – C2C5C1C4 2C8 5C8 5C0 1H0- 9 7 s 6 s 6 s 6 s 8 e V VP ( HH ( HH ( HH ( LL ( LL ( LL ( LL ( V 6 Ph 4 0 5 1 T b G-9 a 6 9 m 9 0 0 1 1 1 0- 9 2 3 4 5 6 7 8 9 1 1u N 1 1 1 6 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 4 2 0 2 . DT 2 I . DFY S S1 3 I Y I T A A S S A A SGWT SGWAT SGWAS L L AL L ALGCD ALGCD ALGCD 2 C I A T T T T TG AG S GMYEA TGGMYE TGMYE A GG SA GG SA D WT R G A G DR GG A G DR EY RG LRG LR LVG I YV LTVI YV LTVI Y I K LER VI Y I K LE RG LAL V I YVSS RG LAL LTVLN I YVTSS RG LAL VLN I YVTSS VLN GY LKLKLKTLKT LKDFM L LKDFM L LKDFM KGG KPTKPTKP KP KPTGQ L KTGQ L KTGQ L PR PGTPGTPGQPG PGSY PPGSY PPGSY AA KGKGKGKQG KTGAAV T T KGAV KTGAV QC RF GPFGPFGFGF GFCN GFACTN GFACTN VYV KT QNKTPKTPKT PKT SQNSQNSQNS QNSS LK P S KT QNSS LK P S KT QNSS LKS WAS QSQSQSQS QSR LND QSR LND QSR LN C YDYDYDYD Y S S SD MD Y Y DRK YDRK YDRK YE W DA ASWSWYSWYS WYSGI S WYS I S WYS I S LQALQALQALQ AQPT AQGPT AQGPT ASR L LS YTCYYTCYYTCYYTC L Y YTCYQF L DN VRSYCQF LYCQFS GSTYVRS GSTYVRGS FM I GY STI GY STI GY STI GY ST I GYL STGKAV LI GYL STGKV LI GYL V STGKK GSQ L QA SFDQA SFDQA SFDQA SF A D QSFG DGWV L A TQSFGA DGWV LQA SFGA DGWV L AY AV AEAEAEAE AEST CT QPQPQPQP QPEAYGAESTT QQPEAYGAESTT QQPEAYGQ SLNK C ITQ LC ITQ LC ITQ LC ITQ L C ITQ L V LY QTGC ITQ L V LY QTGC ITQ L V LY QTG RLSN TS VI STS VI STS VI STS VI S TS VI SENW DLTS VI SENW DLTS VI SENW DL G SD R RK DT LRDT LRDT LRDT L RDT LQSDTR GI DFYDT LQSDT I FYRDT LQSDT I F K Y A GPI ST GT VFGT QF DVFGT DVFGT VF GT VFGI Y I TGTGD VFGI Y I TGTGD VFGI Y I T WS VLRGSS SATS SGATSD SSGATSD SSGAT SD ATG GCA SSGS SGSSI AGSD TATG GCA SGSSI AGSDGCA TATG SGSSI AGT AY GKVL GAKSGL L L L GWY SSGSGSG SGGE LVLSGGWY GE LVLSGGWY GE LV C D M TD 3 8 G VS SSSSSS SSGGGYSSGGGYSSGGGY Y D SW ETLTPSGSPGPGPG PGG VA FSSFSSFSSF SSFSKGPSGSGSKGPSGSGSKG SS SS LP LYGQ Q QY TRQ TRQ TRQ TR Q TRGG GPRQFGG AATRGPRQFGG AATRGPR S AA S GT AS ETGLS QPLS VQPLS VQPLS VQP LS V QPVGQCFLS QP CLS C F VGQFQPVGQF S V G QN DW LF A GF A GF A GF A G F A GG SR VY VF A GG SR VY VF A GG SR VY V F G M C A R ) G ; ) ) 8 G K6 ) 1 T_ 8 p 6 8 6 4 G 6 ) p a a C S _ S ) G _ G ) ) g n S1 S _ C a a p 0 G p p a 7 6 7 6 g in g r in r i r eb 1 7 T 6 p A a ( K _ 5 5 A ( _5 K ( _) ) _ A _ GH HL ) _ L) e s b e b mu 5 8 6V-r VV ( HVV ( K n e5 -r e_ 5 -r _ 56 e c mu m 4 n n u n o 1 e o o H G 0 c 0 c 0 c 0 c S _k 0 c k 0 c e k 0 c A L u q H H A; s s- i c 4 0 s- 40 s- 40 s- 4 -Gi 0 4 0 7nl- s- 4 i nl- s- 4 i nl- s- 4_ S) 7 e s A ; A ; 8 31 tu G- G- G- G- G6 -AL0 ( V ( G L0 - V ( G L0 - V ( G19 - 0 1 6 3 d e 1 2 4 H- 2 6 7 H- 3H-Ce P p 6 7 a 0 - 6 0- 60- 60 - 6 0_ - 5 0vF6 0- vF6 0- vF6 0- T _O s R a R b DC7R 2D7R 5D8 01 7 r H9 L9L9L9L9cc9 c9 c9H8 P – HCHC e V 6 V 6 V 6 V 6 V 6 s s 6 s 6 s 6 V ( 2 H ( H ( HH (Ph 5 1 T 0 b E 9 a -1 9 0 1 2 3 4 5 6 7 0 - 8 9 0 1 1 m u 2 N 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 9 6 2 1 2 1 3 1 4 2 0 2 TG TG S SA S SA SGYSL SAGYSL. 2 V. 1 MW V L MW A L LYG LLGAL L ALGSSN LGSSN VYLVYI KYI K YLVS YLS 3 C IAP C IAP STGTSEGE STSM GVTSM 2 A WS AS AVGVAVGV AVFSQ L GVFSQ L ELG W A E LG AK A I YTA IYKTA IYKTAK GI YT AK G I YTFY AK GVT I YTFY GV G GR L LGTL LGTL LQL LQ L LGS L LGST KR GA PC KGAC KPGKGKGKG KTGANK KTGANK AFY PF PFPPFPPFPPF PFACS PFACS QV AYV KA GFVKA GFVKA GFVKA P GFV KA GFVS LN P D KA RA Q K GFVS LND VT RA PFPFPFPF PF VS KS R L SKSSKSSKSS KSSI S PFR T KSLK SI ST WDE WDE QQGQGQGQG QSGGF QSGGF CA CA YDQYDQYDQYD QYDGR Q G YDGR MYR SL MR S YSLS WY ATWYWYWY WYP SATSATSATS ATSQKA WYPG ATSQKA SSN SN L SM SSM NQL LCNQL LCNQLQ LQV LW LQV LW LCNLC NLCGGS NLCGS FSQ L F FY SQ L I GYYTI GYYTI GYYTI GYYT I GYYTGAS SYSI GYG YTGAS SYS GV FY ST GV S T QASASASA SAEDT SFQSFQSFQSF QSF DV LS QAFEDTDV L AN SN ADADADAD DV LDV LSDV LDV L AK AAK QEQEQEQE AQEQSTAQEQST CS SN CSN CP LD S I TQ LCP ITQ LCP ITQ LCP I Q L CPE I Q LQSGGQCPES I Q LQGGQ RLK LD RK TVS SI ST LI S I STVS ISTVST I TVS T I T ST VS I GVGTTSSTG WVI GVW GF ST RDT LRDT LRS DT LRS DT L RSG DT LGMCLPRSGM DT LGCLP GR GFR GTGTGTGT GTGI AAGTGI AA PG GPG VFVFVFVF VFSGWSVFSGWS QK QK SADTSADTSADTSADT SADTGEGSDT EG A VA VA G G G G GGLAAGGGLA A F L L V GWS L S GWS LSSS GLSSS GLSSS GLSS S G LSS GRS GGKALSSGGKRA 4 N F 8 L S S S G S GA SYSG DSGA S SYS DSPSS VSGPSS SSGPSS SGPS S SG PSSGGPCFSGSGC SGGAYPSGPF SGGAY IG A TV T QFQSFQSFQS QSGQVQSGQV S A L G EDLEDKT T T TF TFGRATFGRA Q L Y D V L V L V L V LMRMR R R RSVT RSVT S Y G YT DT DT S SMSMS MSGWDMSGWD A S S QESSGQS QSQSQSQS QS CEQS CE Q A A G A Q Q G QE QS GG QV DV GV DV GV DV GV DV G V DV GG G MA RV DV GG G MA R ) ) G ) K 6 ) ) 8 G ) p a 6 ) ) g n 4 1 8 6 p a C S _ gn i r g ) n i e i r g n r T e _ S S _ C a a p G pa 7 6 b e b i r e b 1 b m 0 G7 p A 6 p a K ) ) _ m m m u n 1 n T A _ GH HL u o u n o u n o ( ( K ( o H 6 _ 6 6 _ 6 _ 6 8 6V-r VV ( e6 -r e_ 6 G H H H A; 0 c 0 c 0 c 0 c 0 c S _k i 0 c k i 0 c As A ic ; A ; A ; 8 3 s- 2 t 0 s- 20 s- 20 s- 20 s- 2 -Gn 0 2 0 7 6 l- s- n L2 0 l- s- 20 u 1 2 4 L- 2 2 7 L- 2 2 7 L- 31 L- E-1 E-1 E-1 E-1 E- E-A( _ V ( E L - V ( E-Ce P p RD 7 a r C4R 2D8R L ( C5D8R LC5D70 LC1 LH0- 9 H0- 9 L0 - 1 1 9L0- 9L0- 9L0- 6 9 0v cF1 c0 - v 9 F1 c0- 9 7e L L ( L ( L ( V 6 V 6 V 6 V 6 V 6 V 6 s s 6 s 6 8Ph 5 1 T b 9 a 9 m 1 3 2 3 3 4 5 6 7 8 9 0 1 2 1 1u N 1 1 3 1 3 1 3 1 3 1 3 1 3 1 3 1 4 1 4 1 4 1 4 2 0 2 SGYL 2 GL GL LGLGLDGLGAD LGAD . AL . 1 YG LSSS GYF GYF DLDL LDLNSEA DLNEA 3 2 GVSN GTSM VSA VA S VFQ L I I G S S I I G AVS S DT LAV KDTG LGV TS KDLAV T KDLS GVS KLR SL G S DT LS KLR SLS AKS I GS I GS I YTI YTI YTI YTFN I YTFN IYTFY LLGGV YD YD L LGL LGL LGL LGGM L LGGM ST E KTAN LGG E LGG KG PGKCKCKCSAQ KCSAQ PG FPGFPGFPGFALY PGFALY PFAK ACS GKH CKH P N GR GR QAPAPAPACV PACV D GTQGTQGTQGTST QGTST KGFS LD PDR PR PDSPDSPDPDLNK PDLNK PV KF K AA AA K SR L SKSKSSKSSR LS KSSR LS SI S QRC QRC QSQSQSQSSN QSSN QS TF VFY VFY QYSQYSQYSQYSGD QSGD QGGGR WT WT F F F FG WGWG G GPR YFGGPR YDPG GA GA SGSGWSGWSGQI S WSGQI S WY ATQK MTD MSD LALALALAV LSV LA AE AE QCQCQCQCLTF LAVTF GRSQCL GRS NQGW NSA NSA NYYNYYNYNYGGSNYGGS LC G GS SR L SR L NTNTNYTNYT KVNYT KV I Y SYTGAS L L SYASYASYASAGSAKSAGSAK QASYS EDTV SS FN SS FN VSFDVSFDVSFY DVSFDE SFV LDK GM GM EEEEEEEEVW LSV LY ATV GSF EDE EVW LSV L ATG AD DV L SQ L SQ L SPSPSPSPQYYQSPQYQ QEQST AY AY S CPES I G A T V AV QQ LSQ SQLSQ SQLSQV GSQVYG SQLSETFWQLSETF TQ LQSGTQ CT CT C G S VSGV LNK S LNK I TTI SC TLI TTI SC TLI TTI SC TLI TTI SS TGGLC S W LGGYFI TTI S TGGL GGYF RI SGMW R LS R LS VTVTVTVTGVAVLGVA DT GLGCL SN SN R VTFGSI AP FRFRFRFGSI AS GD GR GD D GR GDD TGDD TGDD TGDSI GRT TGI GSDFGSI SGDSI G TGI GSS SADTGW EG PI S PI S VGVGVGVGGYDVGGYD S GLA QT QT SSSSSSSSGE LGSSGE LG LGGGR V LF VFSAGAGAGAGGCGAGGCG SS SGGSKA RSLR C GGSGGSS LSS SGLSS SGLSS SGLSSK SGGGHRS LSGSGKGHR 5 8 PS SGGG GPAFY GKV GATGKV GAKSS PFSS PFSS PFSS PFGP GADS RSS PFGPADR QS TFGQV S RA EWSV LTSEWSV LTSR QSSR QSSR QSSR QSGQASRGQA SRCQSGRC MRG QSSVS GWD V LAYGQV LAYGTPTPTPTP VFTPSVF QMVMVMVMVGGWYMVGWY VS DV GGCE G MA R) QYGQYG n V ET F VT I QGSI QGSI QGSI QGSGGTAI QGG SGGTA ia W E F W D A D A D A D A G M S D A G M S mod ) y K 6 ) ; 4 C 1 G m 86 m 1 u T 4 1 S_ d _ T ) ) ( S - C C ) G s 1 76 n 0 G8 1 4 1 4G8 i s 1 T ) 6 S 1 T 1 T6 ) A a e H_ m L o c _ ne C1 C 1 _ G _ 7 C _ S _ 1 C1G V-r V ( ) d u _ q 5 4 G ( 1 T 6 5 5 7 ( A ( G ( G ( 6 6 K 6 l e e s 3 _ 3 3 _ 3 _ 3 _ 3) _A ( 3 _ ) G e k A i s n0 c 4 1 b a i l- s- L 2 _ S) r d 0 16 a e s c 0 s - c 0 s - c 0 s - c 0 s - c 0 s - c K s - 6 0 4 c K s - 6 4 t 9 v y a b 4 0 4 0 4 0 4 0 4 0 4 1 4 1 i c L V0 ( E0 1 6 d – H H H 0 - H- H- HT _ 0 HT _ Cu e P p v-7 a F1 0- T 3 o _O bi t 3 0c -1 - 0 - 1 0- 10- 10- 10 - v-1 S 0 - 1 0 v-1 S 0 - 1 0 7 r 8 e c s 9 6HR VP ( n a s- H 4 V9 1 H V9 1 L V9 1L V9 1L V9 F 1 c s 9 11 F T ( c s 9 11 T ( Ph A 5 1 T b S 0 9 a h- H 9 t i - 1 0- 4 5 6 7 8 9 0 1 1 m u 3 4 n N 1 A 9 1 4 1 4 1 4 1 4 1 4 1 4 1 5 1 4 2 0 2 . 2 SW GL SW SGQ SGQ VT SQ VHT V V V D GLD VG VG HG G W SQ G SG GS G G . 1 G 3 2 I YG G CG I YG I YW I YF G G CF G VWF GQ A SA SA K CK VW I YCK V I G L L L IGW C WG I GW GF GF G MY MY I YCFK GFY YCWF TS TS TS ES WL L I F ML GKF A A A LY W ES LY E LV WV S E LS GY WV L S WL ES WY R L I Y I K R IYG L RG V I Y LV GW CW GS GG E LV LS E LV L LE L LT LT KA GS KA QG QS GS GGS S KVK KVK LKVK PR GSR GST GT KSG KT GRS PT PT PT AA PAA PA PA G C GS PA GGS AG AG AG QC QC AQC AQY PT MCY PT KGQ KT KC RY RY RYY RY PA QY SA PGF GPGF GPG VYV VYV VV VV QCY RM QC KD KD KFD WA CT WA WA A T WT I RY VA I RYY QG QG QG PD CTD CD CD WVA WTD WV QY QY QY YE PYE ME ME C C MT MS CAT YN YN YN YA NAR YA YS YS L NR YR L YR L YD YA YA M YK YD WG YE ALY WG S ALY WG S ALYS SS AL NSS NSS NA NLS NP VS VS VS FN SSN NS NS L STV SS STV NQ NQ NQ SM FSM FL FE NS NW NS I NC I NC I NC FGQ F I DE I DM FS L F Q FN SYY SYY SYY SL AY GQ SLY GM SY GY I D I D L I D L QT QT QT A SA GSK L GSYA GQ AVT AVT A L SA SA SA CN ACN KT AT VS CS KCI S TF GKT I S IS AS AF AF AF F QD QD QD SLKS S LKS ST VS SS E VT CQ D T LN CS CQ CEP CEP CEP K I SK T LN I T I T I T R LN RN KD KR S SD LSD VA VT S LT K LDA S LK TVQ L TVQ L TVQ L TST RS RS RS GR GR SI T SM TD SI S D DI S DI S DI S GI S PT GI S S PT GT A PV GT EV ET Q PV SPI S GV SPP IN GVT L G T VT L GVT L QF VR QFR KR KR KT PKQ KI SF STF STF LG GK V LG KG KG K VK VVR KG V A VKA LG VK L TR AD AD AD 6 8 G S LT S LT S LT GTSGTSE GWSG NVGWSA E GK EA GPK SG SG SG G L NVGW A SNSGW P SNSGA AW S GNSGSA SS PG SS PG SS PG VT . e c SATSAT AS AS W SAS W SS SS SS V noi t n e EDV LEDV LQ u V LY I VYYVQ TVYYVE T NASQYYVQ T NASQTGS QTGS QTGS KT TTGV LY ITTGLTV LLTV L Q LYSVV LTV L Q LYSVMF MF MF G n q e QVDQQVDQQVNVTQVNVTQVYTTQVNVTQVYTTI QRS I QRS I QRS T ev S E S G E S G Q H G Q H G Q NV L E H V L G in G Q N D P D P D P F tne s t o e r e t o e t o e p r e r e ) r e ) 7 e h h d t e h d C1 h d 1 k o i n ei n 4ei S ( t f e f e 1 ( _nf e 4 de t d 4 i l s a ) W d 4 F is ) W d RF F is ) d er o s H) 1L- λo H) 1L- λo H) 1 e s a n G e A c ot i ) 1 A ( W s n e i C V _4 F_ 1 A ( W F_ 1 A ( W Fb- 2 L e o V _4 2L V _4 2Lin l i p 1 c u t Cu q e e r c 5 ( 1 a pp e u t 1 a i g n p p e o u t 1 a i g n p p e o u t i g n m r e s s e 3 _ H 3 a H 1 b H 1 H 4 H 5 H 6 d H a k s el o d ea k s el o d ea k s el o gλ P p D V V V V V V V Vr ( b Vr ( b Vr ( e b V7 a 7 r r e 8 e Ph h t 5 1 T O b . D 9 a 2 I 9 m l e b QE: O1 5 2 5 3 5 4 5 5 5 6 7 8 9 0 1 1 1u N a T S N 1 1 1 1 1 5 1 5 1 5 1 5 1 6 1 6 1 4 2 l 0 a . ) 2 i n s r e GSRETS LA SGS AS. 2 gi r t t SAGGSPASSE GSGAD YSNP . 1 3 2 oe l SG o GAPNYVH M GGSQSKHVP LT t i c F l RGWC SPQQNV SFGTNPEC eva it t i SPS lad VGAPEEKS TC RGGESVGVQ GYC YTRPMSF SPGGSSSVDVPE en r s a G nl d SG EGTH STKE TEV V N GSGKYR o LSSK YGADKR GG SACG SG PQ AGI YV LWP STNQ io b GS N Q I LG GFG ta i n c i GGGI GSKHVP L I A TW GS QKVKA if n RGSY dw oo I Y h L LGI GPEEYR GRGWG GEWEPK GWSV SGVS DQKD I Y LGL GGLDI ST KDEK m ( s ne K r PGE LVVPV L TH o PGCK E VYRT L KPG LGPASVI EP it a KG LKGLTWPQK PVAG A KTQGD ns e e G vu PV i TP d K AGNFGS V GVKRYV DVP LA QVQQGKVKALF PKTV GWVVCK 8r i n o t s ne r QKTR YGVI WEPKFS QTI YYGTNS 5 7 , e sR CWI D W NDI STG QYGFWRVEVK SGTYE THKDS WAYA DCPTC r 6 o , 5 e r C S p 4 , e; VF d WAMASSI ED GYGVPL AN LPSFYRKY DAV YI GNFVI SE , 4 3 , G he , 3G t n S ol i I SGSRAVP SYN TLCVLP TTSA APT I GT SM SFDLK TG 2 , G t r e QS ,1 2 = , 1G I DFV S G S gdn SSGYCKT DSEDN L VTNSKY QSSAAKPW =G i n u AQDTSAVEVN AYFAR LKD n nG d rod V n CSASDPKN QNC SSNPPQ in G L e r i nG erG c c a I VGG L G L G L G L G l TQCETC LG L e hSGe hSG a, s d TC KEP CSLM o VYS LARYQ I TTQCR L F L H LV 7 8 RYTR T L R LI S MEKG VYYSQ LF YVT LV LI LI LV TV LTV VTA LT LVV TLTw V,) nGw G,) nGG i ed bn D S o i GASNLTGNS RDTGGVS VFGMDN TT LE GAPTPS GV KQETKTQTQTKTKTSG GSSG GS bi t nw SDEGPQKPWWEGPVF SDQNKGVR GGGGGGGG G G no APQLKD AEV LSL LY GGESGGTCGGGG ah S L Q Y QVS L P NET GGGGG GGGG s L V LVPP I ASS L QGDPS F F F F FG FG FG FG ( GG ( G i c i f c e r SSS T e I SGNF L H LDSLSSSLG SGK ISAPNY p a i , s t PS n e c QT LGGKSFV TSNVT SLPKPI SSETCQ VYFQTSQT L VFPPEE )2 T l 1 C 1 tue s n e r e T u MFEVDPSGYTTLR QGCR QDTVKGVVKHMFQQTP 1kS 4 ( 1 4 G1 mp q G 7 4f i e S DV DG SQI S QTG FL LRV YAN CH LI QD DT GSAH G WTK KT K R4444444 , B 4 __ DCs e - FR dFRFRFRFRFRFRF xc 2 Bc c eddddddd 1 n e R I F- F- s e s eeeeee Lu L L l e b o a ssssss D baa aaaaa P- q e _ y o h n k - G eb-eb- b- b- b- b- b - i t s e m_ 3 _ 8 A in l in e in e in e in e in ee n a c m0 c 1 c si c m l l l l l in l in l 1122 r m r m r m r m r m r m r m r L L L L fon e r n u o ds- 4 s- 3 t e g e g e g e g e g e g e g e g r e r k e r k e r k e k s e l ef e t i ip _ tu 0 0 oB- B-Cu e P p λ Vλ Vλ Vλ Vλ Vλ Vλ Vλ Vn i Ln i Ln i Ln i L p r 7 a m. e r . c h 6 0 i s e i t 2- 2-7 r 8 e a , D w _ 0 7 7 3 Ph xEs : e c 3 7 5 1 T b 9 a 3 n e D I 9 m 2 6 : 6 4 3 64 65 66 67 68 69 60 71 72 73 7 l e buq QEOOR4 5 1 1u N 1 1 1 1 1 1 11 1 1 1 1 a Te s S N P 7 1 7 1 4 2 0 2 Q GSRETS LA WVSSVSN STE GSRETS LA VS. 2 Q S. 1 W AGG SGSPSSE SSSEVN GGC SAGGSPSSE WSSS 3 2 R FGAAPNVH APPPKCE SSAY SGAAPNVH SAPP S WCYQQM YQGKVTKP GK LV FGWCYQM YGKV K RSGSPENVS TTTVRYQ SF K RGSPQENVS QTTV D PSAPEKTC NSVI SE V VG KG RQEH SPSAPEKTC TSV L GYCTRMS GASM LGNS SDKE VGYCTRMS NAS T GGYTHPEF SSSS LT EKPSY GGGYTHPEF GSSS L K SGSTKTEVN SVSDN LWGVPD SGSTKTEV SSVS S E Y LSSKKRG GT L YGAD YKWEPGSP I A ELSSKKRN G GTY F GGSSACQ GV SNPQ I Y LLP TGKDVS PQI A L F GGI GKHP LW SGSSPHDSAF LI LFK FVS SL YGADACQ GV T GGSS GGI GSNP KHP LQ I Y LL TG SGS S RGYPVETR I AQ F LLSSKAPL RGYPVETWR I AQS GD I YS LGI GEVYS WGQ LFVPY RATS WSGVQK E LWVVT LFQTI YAVS I YS LGI GEVYS GQ L WSGVK WWV S LG L ESVDPD GLASVGYL LTL LGESVDQPD E LLA D KPGLKVEV KDPFPSVKHKRS V PGCVYRT L GT Y KPGLKVEV GDPF ATGVV LNPI KT PGCVYRT L KTT P KG LKLTWPK PGHGRCH LPES KG LKLTWPK GAGH P GVGPGN T FQS AGVL LY AKVD GVGPGNFQS PAGV T PTA K KVQQGKG VKV AL Q F RYGETSW LEGKTK PTAQKGV L QYG VDSPANSHPKGSD KVQGVKAF RDS Y Q N QKTR EKF YGVI WPI SSG WVT LPYVMQQQ QQKTR EKFS VVT L E I YACQQVSQGE YGVI WPI SG WYA N WCW GTI ND YETKD YGG WRSPPEN EKCYFATV WCWTI ND YETKD I YGRGS P S TH S AN TSW S SG H S AN Q VFMSSI ED YDC CTRPMFANPE VFMTSSI ED WYC G WAA N SGYGVPAL SFWSHKEVNL Q WAAYGVPL DFWS S GSRD VP NYVVTTEGYTI GTS SGSRDAVP SYV E I SSYNAV TLCVP LP F VAT SATKKR TVDANSPQ TN I SGYNAV QI G VP LP NVT SSG STLCVAT FA STV WEG VP QSSSI DF GYCKT FDP VTNKY GEEC PSKHVP LWQDS QSI DFCKT FDP SSQ L SSGYVTNKY GEEP IAS L AQDTSAVSVN SAAFPETRAYA ADSAVSVN SAF DS SVSEKN AR LYEVCSKQFNN QTVSEKN AR LY SL C PS I TQADPTCE CSDVGVDCSD CS CCERKP SNKKDQPV YK TYS I TQV I TQADPTCE A CCERKP CSD NK 8 FQ LA T VRYRR LI SYEQ L G R LMV LKV CD E VYRT LTVCK T YWC VYS YLA R Y LI SEQG S LMV L 8 C GKY TL VH D SM LN GASNLKN TGS SQ GLYG LKW TNPKRY QSDTQE RTR VG D LSM GASNLKN R TGS LSQ LYG L CH VFG L SD MDN AEG KLEKGVANFGY LGAKR VFGMDN LE GVA PPQ LPWWEGPPTASK QNTPVKFVF EAFSDAN EEF SDEG K WGGTA L S APPQ LPWEPPNT WA S QYKDVS L VKSGKPKSAPRK S QYKDVS LQKSG SE VH LSQ L M SSV LVPQ TP I GNF L H LI A L GH DS SLSGNSND GDTVEI SG TDS L P HKSSQ ALYT L Q SFV SL P SSV LVPQ GTP NF L H LI ADS SL V LNGS SGDT QV PSGKFVPKGRSNSI ED LP NS PI SGKFVPKGKS NS ST LGSVT LYQSI SKI CV I SN ST LGS T LY GI SK KC QTSNSVFT TSY PVST L L L S L QTSNVVFQTSTS TS TFEDPSGYEFSTDVAPQT G TFEDSPSGYEFS MF MDVKGVKHV LRPQVP LPTTFCVQ MDVK VKH RP EEV RN Q G VTV DG SQI S QTGVV FL LR YAN CHQG LV EQA L APTV FG LCA TKTM NK YI QD DTV GSH AT Q VDVTV GQI SG TGVRV V ANL QGQA LP B- D S Q FL L Y CH L Q A F 2 c B F- c -a Rc F c l e F- o h o n b o p p I L L -l e F- b _ 3 (- . 1n k a k _ _ ( - y o h o .1 m_ 3 n ( o -8 n k G 0 c 9 c) T As s- 40 s - 6 9 3 0 4 c 1 s m - u 0 c 1 c _ ) T ds- s - 6 4 K 3 0 _ t 4 0 3 01 i ct B- B- , B- k u K oB B,Cu e P p 6 7 a r 2 - 0 07 2- T 1 73 0 0 C 1 2 - _ ) 7 p hi - t 6 - 3a c w2 - 0 0 2- T 1 70 1 7 8 e Ph S _ 7 3 S5 1 T 0 9 8 b 7 8 6 9 a 3 9 O 4 R6 7 8 OR9 0 1 1 m u N P 7 1 7 1 7 1 P 7 1 8 1 4 2 0 2 . 2 VS EVN S N GTE GSRETS GC SAGGSPSLA SE GQAC SS RTS GSRETS WTHPMEF SGGPSLA SE . 1 PK 3 2 TCE SSA RKP G Y SGAAPNVH GGSTKEV ASGSAAPNVH ISYQ SK LV FGWCYQM SGAKTKRNG FGWCYQM EG F K RG M RQ SPQ LKN EH SSAPENV EKS FG TC RSGYDAS SYS Q RGSPQENVS S SDK PVGYC SNPQ SSAPEKTC TRMS PGI TKHP L PVGYCRMS TGE P E VSY GGGYHPEF VGDVPVTWR GGYTHPEF DN LWGGPD SGTSTKTEV GGSEECS SGTSTKTEV KPWEPSPA ESSKKRN G SGGSV KDVS SGVK EGSSKKRN G PQ P I A LAI FK L SI LFVS L YGADACQ ALSAVDQPD LGADACQ GGSSNPQ TGVKVEV YGSSNPQ FL H LDSLSGS FVPKGPT L GG RAT RGI GS YKHP PVLTW GGV R GGCV LYRT WPL G K GG RGI GS YKHP PVLTWR VT SLY VFQTI YAS V I YS LGI GEE SVYS KGI ATNQS GVK I YSWGFGY L I YS LGI GEE SVYS GVK PSGY GVKHL LTS L LGWS RS KGE LVDQ KVPD EV L LGE KGLQGKVKF LGWSDQ EA PD KF KGE LVKVEV GV LLRV LNK YCH LPPI KY ET P S PGC YRT K AKVD GG LKV LWPLK PGG AGKWP S PGC YRT GLNDI SG PKGKV LWPLK VGTNQS KG LPFETKD GLGTNQS ET PSW LEGKK PTPAGFGV L GVPAHI ES PVTPGFGV A SHPTS KVQQKVKF PTQDSPD L KVAQQKVKLF PNYVMKGD QKRGEAKF KVKRSVDAV QKRGEAKF CQQVQQQE QTVI WPI S QTVYAVP LP QTVI WPI S PENSQGFT YGWI NDSTG QYGWDV PT YGWI NDSG PEKCY S AV WCGTYEKD WTCKVAKT WCGTYETKD CRTFWANS SVFMTHI ES AGMRCNK SVFMTHI ES THPM TKTEVLPE AASS ENYI GQ VPD L LFD S WSGYGDAV YVNYAT CVSY AASS EVN WGYGVPD DALV KKRGTTN GSRAVP LP D GSYPKCNE SGSRAVP LP DASQI GTG I SYNLCVAP I N I DSYTKP I SYNLCV P CSN KHP PQSS S LWQDS QT FVKTT SD SSSI DYCNK QTI I FV SAR SI SYEQ ST FVA G Q KTT SSI DYCNK PVTRSSYQ L ADGVTVSY SYFD EEVCSAFA QTSVAEVN AGGSAM LKGNS AS QDGVT TSVAVSY EVN VGVKQNN CSASDPKN QQAATTDN LE CSASDPKN KDQDCSDV I TQCETCKEP CQKVK WG I TQCETCE KDVPVI TQ YET LTCK TC C VYS LAR RI SY I TCC EQ TYT LS SPWEP KDVS L TC VYS LARK RI SYP 9 8 EQ VWRKVYWE RYTR LL MKG VYT LPQI A RYTR LL KG KNPSRYTQG DASSLGN RAS L KPH LDS L D SM LGN TNFQ KGY LDAV FG R GF VFKAN VDGNT SE DFTL GMDN L GGDESF VL SS GA FVPK VFS DGNT S MDN LE SPVKF KEAKSSD AEEF SE RS APPQ LKPWW V DEPSES VT LYQ SEG APPQNSVFT APPQ LKPWWG DEP HPDI SG P DS L QPK YT S L Q LQ VLYK VPPQV IAS L SS L Q LK VKPSGY SGVKH S L Q LQ VLYK VPQV IAS L NVET NHKSSSLSFV SS T H NP SI I SGNF LLD VSLSSSSL CS I SGSGVRV LN S H SS TP ISGNF L H LDS VSLS YVI EPD LP VSI SNS P S ST LGGKSFVT LPYKQP PD ST LGKL LEYC T LA PST LGGKSFVT LPYKQ TD QVAPQT TLTL LL Q CG TTFSENDSPVSFGTYQTTFSEI STPASW NLSEH QTTFSENDSPVSFGTY TVP VLP AT G TMFVQ MDVKGVKH LC TK NK YI QDVH QTVI SGVVNMDTQ LVRPCYV QQMV MQDTVVK ISGVKH GVVN DT GS AT VDV B- DG SQ QT FL LR YA CH LI Q DG SQ VG KP PE EN KS CDV B DG SQ QT FL LR YA CH L 2 c -2 - 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RYR L R L SYEQG KGGVSVNTCR L GGLFVYS TS DYET LRTAK 1 s ) DTASSM LGNSGSGMGRYTG ESVCVWRPKDFWC Le c s GFGNT PQG NEGKEGCSTDAGP KNLK ATN NFQSG Y V LVD SEQE D VG P- n e r e VD ut t SEGMDL PQKPWWEGLGI A PTSWEG PVF SDQ LKSATSKG PVKFAPKR i t GSGKEAFSQAN n a q e e l AP ic S L QQL VYKDV VPQI S LGG GGE LDGAEV SPL G PDGHPKSLLEF GKSNDI SGSSRS ro sl a l a t SLLTP i SSGNFH L ADS SLKGCGL Q SI YGKLSLG SG SEI ST TSVE NHTDS KSPI SPK ST LYTV 2 Bi n PI SGKLV gdn ST LGSFT LPYKL LKG LGTS S VPWPI SE QLVK S SI CVI ED PLQTTF FNPS Ri r e o a c n QTTFSENV DSPVSFQ GTYPPTA VQTS YQT TLTV L QR VGSPYTDVAV PPPMDN QK AA LQ TV VL AT TI QTL L S L I L Lo t l doe DG SC VG Q x 7 bu MDVK q QTVI SGVKHKKRD FQ GVVNGTVRM I DQ 4 e v Di t n i e S ) V DG SQ QT FL LR YA CH LP KG CW CA RQ DT GS n D G B c F- - Cl a a p x e r wo -1 1 s L e - b nh s Dl o - 1 1 o n o b n ( o p n a Li k k ( Dot a e r P d e h, 2 l eo h E- k, 2 4 4 P - i c a s ) v _ 3 8 1 ) v G 2 c _ ) T 2 i t i f s e a F c -8 F c As s - 6 c s- n af doui dn b- 3 s ( 0 - c s- 6 _ s (- c 24 1 2 omsot 0 c 42 ) c i 0 t K 0k Cu G e - s ( e i B 3, - F- 0 S F- T G-3 C _ l eno rRi pr 0 2 2 ) v B-64 2 ) v P p 7 a 0 - 1 0 0- ) p p mi tnDc s - 73 F c 2- G ( _7 0 F c 7 r e 3 31 S 3 3 a c a x e vC; e d D ( s ( 0 7c Fc s s ( 8Ph 5 1 T E b : i n t en 2 9 a 4nl i D I 7 64 9 6 l e er eQ: O 1 1 m u 5 8 N 1 8 1 b a sdEOR7 Te r pn u S N P 8 8 1 8 1 4 2 0 2 . 2 DASQ. SNPQSG AA 1 S TCN G M SSRETS AGGSPSLA SEQGMA VSVSNGS QFVYC WSSSEVNGFGWG RGA 3 2 KH PVP LTWQSS L RCQR EECSI TCL Q L SGAAPNVHYYYF SPPPKCESSGGWL SY FG CYQM V RGWSPQNVWGN AYNY A TVA YGVTKPG QKTTRYQGPVSV GLNF SVVKTYGT SSAPEEKSCLNFS TSVVI SEGGGGGMG SGDQPDVYTGN PVGYCRTSNGSD NASM LKNGSGCS VKVEVRAPK GGYTHPMEFSSFE GSSTGSEKELGI ATE VY L RT LDFQS WPKG SGTTKEVI GR VDVN SGSA SS D EGSSKTRNSSAR L SVLDN TSKLWGTSWP EPGGED TNQSSELK LSGADKACGQGAS GVY LPWDVS LGGLG GFGY LAPGGI S YGSSNPQSACN GLTGKPQI A KGCKG QKKFS L Q L T GGI GSHP LQATS LM I Y SP DS LI YGGL GVEAKFSSGSF GGYKPVTWQS Q SGSFH LSSL L G LPT WP SSI SER RSI GEEYRCQR LL I AQGQ LLVPKKVAW LDDI STGPTVG I YG SVVSKI TCYSY W VFT LYQPTQTY TE AHKD I SSQLTLN L LGWSGQDTVYGVT EWAVSVFTPVRD FQA KGE LVDVPVRTGN LLDPPSGY KHKKVR GSVEPD LT GDAVMDQWS TSSS P V PG YV GCK KV LYER WPT LDAPK GKTFGV NGTWA KGFQS GATHGVV L HPGCCR DVP LPPQGGAK KLGT TV NQSVDEV LN PGVLRYCLKGMA VVAKTDSGY SGGYTV L GPVPGFG T KTVAQKKV LSAPGD AG GK QYGL SET PSW LAEQQFYC YVYF YC GTNKGSGSNG QKQGVAFFS L Q LGI S RDTANSH YNYV RVSY AEVNGG GSFGG GWQ QTR G YGVI WEPK ISSSSST GSI SEF VVLPYVM R WYACQQVWGNTA NSALYFS CPKN FTCEPGSRSGA WCWI NDETDPTV GGW SGTYTHKSSLTLGN I YGGSPEKCNNSD WRANPERTSSGFE YRK VAI SYQGPSVL EGGVGLN VFM F WAASSI E GVPD LQTFQ DQA YC CTPMF SWSDFWSHKEVI GSGA SRSSR L SM DLK TGNSGG GSGMG SGY GCS I SGSNRDAVMT AVP LP SS PQGGAVSY KNVVTTKTKEN RGQSSA GAS CN EDN LEKE WGLTGSI AT SY WE QTLCVATV SI DFV DSGY CKTSGGYV LFA STVDASQAA DPCNPQQTS LM AK RLPWDEVP SKQ SGG P SS GGE LD ADGYTNKGSG SVVSYGGSTTFEESHP LWCS GNGGAPFKVETRI TQR L Q L SY NPP I A L DS LK I GCG QTVAEVN GKG GS L CSASDPKNGFRGGQSA GWGAR LYP DEVVCSTC V GVKVYGT MF Q L H LVS LF PSY G KL L G LPT I TQ TCC SETC ARKE YPSS AWCSKKDQDRY QGPGSGLSNVKVPVDTG APNKS 1 9 YVT LYQKVAW VSVFTPTQT VYL Y RYTR L R LI S MEKGGV VNLMLDYET LGFQN GGGLFRQCVWRKVDV LD TPSGYPVRD DASSLGN NGVKHKKVR GFGNTNSEGSGMGLL KEGCS G TSYVLKNPSSEGK ATFQY LAPGI S KGV S RV LNGTWA VD CH NL L LPGCR SEGMDL PQKPWWEGLG PTSI A WEG PGTANKGFS PNTSPVKAFLSQ LGSTF DEYTW KP LAK EQCGMA AP FYC S L Q LQL VYKDV VPQI AS LGG GGE LDGQKSGGKE HPKSSSER ISGPI SVG ISASSHQYVYF S LTP PNYVM YNY SSGNFH LDS SLKGCGV LNSND SI YGKLGDTVETDST KSQLLN TQAS TFCQQNVW SAGNV PI S YT GKLV FA ST RPPEKCLN LGSFT LPYKL LKG LGPT WGKSNH GI SKI CSI ED LT MFQ DSWSSV CERTSNGSS Q FD TTFSENV DSPVSFQ GTYPV PTA VQTYSTS VPVQTG EFSYTDVAPVGGAYK GQT AHPKMEFS VI GSRGE SA MQDTVVK ISGVVKVHKKRD RP NGTVRV LGA LQTVP VLP ATDSG TSGGYV TLT WT KT KE RN GS QS SAR AL S ) VGQTG D D S Q FL LR YA CH LP KG CW CA RQ QQ AP FG LC TK NK YG GS GS GN GG Q B-1 -- L - - bo 1 n 1 D-1 kE 1 -1 1 - P _ d l eoE - o b oE-8 8 8 1 e G 1 c-8 s h a _ b 3 8 0 1 n ( n k 1- c-8 8 1c _ ) T8 6 A s- 6 _ - 3 s- 6 _ s - 6 4_ si c 3 t 0 2 ) 0 B4 0 2 ) 3 01K2 ) u B- S 4 -0 B- S 4 B- , S 4 Ce 0 G ( P p 7 7 2 - 6 G ( 7 0 T 1 G ( 7 a 7 r 2- eh 7 3 _ c0 Fc s 7 3 2 ( - 07 _ c0 Fc s 2- 73 0 1 S_ c0 Fc s 8P 7 8 5 1 T b 6 4 9 a 9 m OR9 0 1 1u N P 8 1 9 1 4 2 0 2 STE GSRETS LAQGFMA GQACRTSNGSFD GSRE. 2 GGC SAGGSPSSEQVYC SSWTHPMEFSSGE SAGGP. 1 SSA 3 2 G Y SGAAPNVHYYYF GGSTKEVI GRSA SGSAAP SFK LV FGWCYQMWGNNYV SGAKTKRNGSSAR L FGWC QK RGSPQENVSAYTA FG RGYDASQQSSGAS RGSP REH SPSGAPEKTCLNFS SSYI TSSNPQAACN SPSGAP SPDKE V VSY GGYCTRMSNGSSFD PG GYTHP DKHP LWQTS LM VGYCT KEFVSRGEA VGVPVTRCSR L Q L GGYTH GPPD SGSTTENI GSSSR GGSEE SVCVSI TQSY SGST SI FAK ELSSKDKRCGQSAL SAGGSGQKTCYGV ELSSK A ILF GVS SL YGA GGSSAPQSGA GPT L GGI GSNHP YK LQAATCS SN LSAVDPDVVRYGTN YGADS M TGGVVKVYETDTAPK GGSS RAT RG IYAS S PVTWQSL GGCV L RPL EEYRCQR L Q L G KGFQS GGI GYK KGI ATWNQSVDV LN RGS PE LLV I YGI GSVVSI TCSY TS L L S LKGWSGQK GE LVD DTVYYGVT I YS VPVRTGN L LGWG GE LQF GKG VKY LSAEGDK I YGI GS AF PG L FS L Q LGI S LGWS T KGE LV KR PPI KY PGCKYERT LDAPK KPGGKWEPKSSSSF PGCK KET VS P D KGG LKV GLTWP NQKGFQS SVD AG LDI SGS EV LN KG LGPNFET PI SER P KDSTV LG KGG LKV GLT GKK PVTPAGFGVSAPGDK GVPAHI ESQLTQN PVTPG PTS KVQQKVKLFSQ LGI S PTQDSVPD LTFQAS KVAQQ KGQD QKRGEAKFLSGT KQVKRSYDAVMDSWSS QKRG QQYGQ FE QTV T YG I WPI SSSSI SEF QTVAVP LPQTGAV QTVI W V WCWTI NDETGDPS T L V LRG YGW TCDVVAPTV TDGSGGYK YG V WCWI N WA SG Y KSS W K K SGGYL GTY ANSE VFMTHSI E QTFQNA AGMRCTNKGSSTT SVFMT LPQ WAASGVPD LTDQWSLFVDYAVSYNGGGNGG WAAS YTI G IGTS TN SGY ISGSNRDAVMTS AVP LP SS PQGGAVYNGC KDI DSYEV YPKCNGSG Q EGFGWG SGYG ISGSNRA SSG SY QDS QTLCVATV SI DFV DSGY I NDSVTKPSRSGAW SYLC CKTSGGYV LSTFARYQGPSGVL QTSI DF SSQ SSGYTNKGSGSTTQI I SFSI SEGGVGLNF SSGY AYLA ADSVAVSYGGGNGSYGDM LKNGGGMG ADSVA QFN QTVSEVKNNGSFGGQAQGSAATGSGSGCS QTVS CN ITSD CSADPTCEGRGWGCQATDN LEKGELGI AT CSAD TQV I TQ CK TCCE YS LARKYPSS QGPGAW SGVLNI TQKVK TCYCSPWW DEPTS E I TQCE V GGWP TCYS LA 2 9 VRY DYWC V TQE RY R DTR LLI SEGGVGLFVYT SM LKNGGGMGRTL S S LKPPQI AS L SLGGE KGLD V CG RYTR L R LS GAVG AS VFKR G NTGSGS VFGMDN LEKEG LGC I STDAL K GFTLSF L H LD VSPSKI YGKG LG LGL DAS T G N VFGM SDAN AEEF SD S AEG K WGTSAEVD PPQ LPW EESVFT S DEPGGWPSPPQVSLY VFQL TKVPW SDEG PTAT APPQ L LPRPK) YTDS L Q LQ VLYK VPQV IAS LGKGGE LDA KN CGS L Q L VKPSGY GVKHPVQ KKRYD) DS L Q LQ VLYV SQ LFVBSS TPH LDS LI YGKGSSLSS VVNGTVRBSS T SSNP -1 SI SGNF LVSS GLSI SG GRL HPGWA-1 SI SGN PI SNS L PS GKF PKL L G LPTP PDKL LYCLKCCRL P GK STLS L D T LGSVT LYQKVAWST LG ETWAQGMADST LGS QLL P QTTSENSVFTPTQTQTSI SPSLEQFYC QTTSN TTCG MFVDKPSGYPVRYTFETVANSVHYVYYF P FEVD MFVQd e DTV GVKHKKTVDMDTQ LRPY M NYd eMDTVK IQDTVHs QV QI SGVVANG RAI Q GCQQVSWGNVs Q QI S D GS AT Va b- DG S QT FL LR Y CH LP KG CW C R DG SQ V KP PE EN K CA LY NT FA S a V DG S T 2 b- Q F 0 2 - a G- - b 0 e - G- - pp 3 0 l - o - 3 h 1 , l e 1 o n -b1 3 E k, o1 0- - 3 l e1 1 ak 3 2 ) o h- 2 n k E- 3 o h E- (8 h v _ 3 8 1 ) v _ 4 8 1 h _ 3 8 1 G 1 c i t F A w c s( 0 c-8 F 6 c _ s ( 2 c- s s- 2 c 7 -c s- s 8 6 i t _ w0 c-8 6_ i 3 t 0k u B-0 C 6 4F 4 0 2 ) -c - F 2 0 2 ) 7 8 s- 6 4 0 2 ) _ O-2 ) v B-6 S 4 - G 2 ) v G-3 S 4 4 OB-6 S 4Ce P p 7 a 2- ) p RP F c 2 - ( _c 7 0 F c 0- G ( _c 7 0 RP 2- G ( _7 0 7 r 7 3 a c ( s ( 0 7 c s s ( 3 3 c s ( 0 7c c s 8 e h 2 F F 5 F P 1 T 5 5 5b 0 5 0 5 9 a 9 O2 3 O4 1 1 m u 1 9 R N 1 P 9 1 9 1 R P 9 1 4 2 0 2 . TS 2 SLA SEQGMA WPG . 1 N QFVYC SFLSCK TNK G YG GGQ GVH GSRETS GSGWG SST SGGPSLAQ SEYGC 3 2 YVH QQMYYYF AVSSVSVNGFRGA GAVE ASGSAANVHWG WGNNY ENVSAYTV YPSEKNSSGGWL STC FG P A YGPPTCEGPSVN FGA RGWCYQMSF SPQNVLA EKTCLNFS I TKVTRKPGVGLF RSY SSAPEEKSCQNT RPMSNGSD DTV KEFSSFE VSSAVI SYEQGGGMG SPK LV P KG VGYCRTSND NGS S EGCS VQKH GGYTPMEFSS TEVI GRGSA SSM LGSGLGI ATE GEK SGTHTKEVYS KRNSSSAR L GASS LT EKTSWP SDE EGSSKTKRNVS ACGQGAS VVSDN LWGGGE LD ASPY LSGADACGSEF NPQQSAACN VTY LKPWEPGGCG LTPD YGSSNPQQSG HP L TS LM CV L KDVS LKGKG GI FAK GGI GSKHP L SG VETWQCS Q I ATGSPQI AS I YGGLT GF GRGYPVTWQAC VYR GVSKI TQR LL WGSPHDL L TCYSYV DQDVYGT E LQGQ L F LLSSL G FVPKKLP VAW KVS L QPTQTY I YST LPL I YSGI GEEYR SVVSCY T L LGWSGQKDI TTYT VPVRTGN GWVVT LYFTPVRD LAAS KGE LVDVPVVA YER WPT LDAPK KLASVGYKK KGFQS GPNPFPSVKHGTVR KV GWA PATS L PPG GCK KV LYER WPT LRF KDDE NFQ KGSVD VSEV PLN GD PFATGV K QDHGRV LNP HLKCCR KRSY KLGTNQSGVP QGMA GI KT GPVTPGFGVSQ VKL EAFA FS L Q LG GI S RSVL LYCA T VYGETW LEQFY YVYC NF PES KVAQKKLAL Y KVKD QKQGVAFFS LS PKSSSSF WASPSNSHWYNV QTK QTRVI WEPKSSI S DI S ETGSI SER CDT LAPYVMAGTA QGSD YG I DI SGST L HKDP I SST QL V TLG MKA QN DR CQQVLYFS YQQ WCWTNETDPT AGPENSNNGSFD WGE SG Y K FMTHI SSF SVEPD DALTFQA VMD SYCSPEKCSSS EA AFT VAASSED V TSWSGYNCRTFI GRGS LYVV WGYGVPALQD VTTG VP LPPQVGGSVS VATDSGAKSYWTPMVSSAR L DNS FVSHKTEENQSAS I I DE S ISGSRD YNAVVP LPM PI QSG CKT TNKSGGY SGYTV LSAV TFTTT VKDKARGSGCN N SQAASM SDQ TI I S STLCVATDS N QSI DFCKTSG VS EVYG NGGS SGNGGGD QSAP AECSNHP PQQTL WCS Q QSY SSGYVTNKYGS PKNGFGWGATPKVLTRI TQR LL AGGS ADSAVSVNGF CSYV QQ QTSVSEKNGR TCEGRG RKPSSGAWKCVFYPECSTYGT CQQ L CQADPTCEGSP ISYQGPSGVL S NT L S LDEVVKV KVKGQDRYTGPNK I TTCA I T YN TCCE YS LARKPS ISYQGV 3 G 9 ME LKGGV NGGGLF TGSGSGMGS LTK L V LKD DVPVD YET LGAQS VYD V VFV LN RTVK RY DTR L R L EGG SM LKNGSA DKN LEKEG LGC IASTESCVWRKSDEGDK DAF C GAFSNTGSGL ESVG PWWGTS LKNPSAPG GDWE VDGMDN LEKD KDEPGGWPPQ P DKNKATFQ ANKGY LFS L Q LGI S VEQ STF SAPVG) SEG KRDAPPQ LKPWWGS DEPA PQV IAS LGKGGE L CGV LSTS SGPVKFS KEASSS PI SER S L Q LAENB F -2 S L Q LQ VLYK VPQV IAS LD IY F L H LDS VSLSI YGKGG GLPDKG PK SHDI SGDST L V LG QN SSS ISRS BSS TPH PKRSI SGNF LLDS VSLSL L F PKL L G LPTG TNVETSQTTQASP YTI L PS GKF PKK VT LYQKVAWSI SSNHK FSWSST LFVL T LGSVT LYQP SVFTPTQTETK SI ED LMDGSVQTNP r QTSNSVFTP PSGYPVRYQ LVSI CVPVQT AKTFNS oTFEVDPSGYQ GVKHKKTVD RSYDAPVGGYVMDLS L f MDVKGVKHG GVVNG RQVGPTVPPDSSGYLTI QTL ni QT I SGVVNP L LR YA CH LP KG CW CA R QK AA LQ TV VL AT T GG SG ST N G DG SC VG Qa VGQT m D S Q FL LR YA CH LK Q od - - - y e - ob1 1 a p n p ml mo h -l e4 0 ( oE n k-8 a k u , 2 o h H- 4 _ ) 1- ( 4 dh ) v F _ 3 1 0- G 2 c T A 6 8 6 9 _ 2 c i t c s( 0 c 1 s s- 4 1 i 2 c 2 0 t K) s- w u G-3, S 2 0k 2 7 -c s - _ F 2 - 4 0) S T 4 G-3 C 6 42 ) B- 4Ce P p 0 - 1 G ( 7 7 a 0 _ ) Ov 6 2G ( 2 7 r 3 3 0 1 8 e S_ c0 c - p R s 3 3 a c P F c s - ( 0 7 _ c0 c s Ph F ( 3 F 7 5 1 T b 9 a 6 4 9 m O u 5 9 6 R7 1 1 N 1 9 1 P 9 1 4 2 0 2 GRG. 2 GS S STE GSRETS PSA GGC SAGGSPSLAQ SEYGW CGG GQAC G SSWTRTSNSL HPMEFSSS . 1 S3 GVGY SSAY SGAAPNVHWGMH GGSTKEVYSSF 2 GGG GSGY GSK GTSSFLV FG K RGWCYQMS SPQENVSLFAR SGAKT QAND FGYDKRNV ASGSFGS QEGA GASFVRQEH SSAPEKTCNTSSR RSGSYSNPQSGA GLGGGKSDK PVGYCRMSNDSLA PGI TSKHP L SAC KD GSGV AGI SV LPSEY GGYTHPEFSSSCF VGD TV VPVTWQCS GPD SGTSTKTEVYSFY GGSEECRSCYL P SLD SI YGI SI GGSPA EG QAI FK LSSKKRNV GADACGSSGT SGGSVVKI TYTR L EFSAA ASASGQDTAS LLGYGI LFS L YGSSNPQSGAS LTGVVKDVPEVVRFG SLKGE KPGL GVS GG CWLGPT L GGI GS YKHP PVLQ TWSG QACD CSEA GGV Y GGCV L RT WPLKDDG GEP QPGKYRAT R TYQG LGFI YAS I YSGI GEE SVYR VSCYL KI TYR L R L KGI I AT YSWGNFQS GYVPQ SQ L V L HG NPVPA KT LAGL QSLVTS L L LGWSG KRS KGE LVDQDTTSS VPEVVAFGN L LG GE LQGKVKL AFAS FS LI SGG HLQKRSDPPI KY PGCKYRT ETS PK AQTV KVD GKV LWPLR KDDGM K EPQ L PGG AGKWE LPKSS DI SGST LGS EYG HWCWGGGKK GLGTNQSG PV VPQY KG TPGFGVSQV LV GLG VPN PFET AHKDSPT SFE DV L MSGGHPTS KVAQKKLFALGT PTQDSI EPD LQTTQ VLFMRKGD QKQRGVEA S KFS L GNK KVRSVAVMGV SQAA NTNDQQQ SRQGFE QTV T YG I WP SSI SGS QKTVYD WI NDI SGST LSN QYGWAVVP LP S PI QGES CSND FSSSS LAY C AV WCTYETKDPTED TCDKVAKTTD FWNS SGFMTHI ESSFV LR WGMRCNKSSG GGSG VYSSFYALPE VAASSPD LQDT I S ALFDATSYGFG NVS GS GTYI GQ WGYGVDAVTGQVT YVYCVEVNGRGS QEF QSGSAT SGAS ADI GTS TN SGSRV G I SYNLAVP LPM PI QSGEF SR DNG IN I DSYPKN SYTCEGS PSPVGG WQACESS RCC AQDS S QT CFVAKTTDSGG SDFVARKYQGGG SSI DYCNKSGGKA QTI I SSI SEGGSG SKI TYYS L DTTR L R LSS SAYQ FLA AS QDGVT TSVAVSYGSG SYFD EVNGF GRGWS AGGSAM LKGNSGGALSG VVAFSNQCNN SD CSAS V I TQCDPKN ETCKEPGSSA QQAA SPGY CQKTT VDKN LEKD WGSG AG TR LDDGMI TQK TC GEGQ LTVC C VYS LAR RI SYEQGVGY I TCCSPWEP GGGGTSTVYT L SKDVS LDG 5 S 9 KSVPP YLS QY AQ L V LVRY TDYW TQE RY AVG DTR R GALL FSSM LKGN GNT SGSGF ASGSVRYT LPQI A EGKLGGKDAS FLTK LPH SF LLDS LI Y L VSSLG KGG FFS LS ISGNG GKVFK SDAENF ) VSDEGMDN L PQKWWGDGVV PSGSI I LG TVDE SESVF PKPG QVT LYQP SS GST LGSAERS DAP SNSPPK B- S L QQLP YKDEV A VPQ S LDGSI GGQAPP SQKNSVFTQG KPSGYGLV DPT SSFEDL QYT 2 SL V LTP I AS I YGYGLL V LSGVKHPT D LQTD RSLFV B TV L QI SSS TPI SNP RSS NS I L PSI SG TGNKF L H LD VSLSL LGE SFTPKKGLWSSS PGCKLPI SGS TPDGVV KL LRLNKL YCH LQK AQT VMGVFSTLS L L QLGSNVLYQP YFSLGI SETW LEYG PI QSERQLLG r TTEDSVFTQG LGPAQTSTPSSH C PTDG TSSSGTT GKMFC oMFDVKPSGYGVA TFEVANYV WSG K YGG GS FGA G WI QDVQ f DTV GSH AT n V i a QVTV GV DG SQI SGVKVH QT FL LR YANP CH LKT LQGSMD QK TR VSI QTQ GLRP QGCPQEQM NVSLF QAT m D D S V K P E K C N D od -a y e b p p ml o -- a m 4 0 o n -b-4 0 k( u h, l e 8 d 2 ) v o h H- k, o nH- _1 2 0 ) v k _1 0 G 1 c hi t F c 3 As s- ws ( 0 c-9 F c 4 2-9 s1 _ s ( c s- 1 _ i c 3 t 0 2 u B- k 5 -c - 0 C _ 0 5 F-2 4 0 2 ) -c 2 ) v B-6 S 4 F-2 0 2 ) )v G-3 S 4Ce P p 7 a 2- ) O 2- G ( 2 0- G ( 2 7 r 7 p R 3a c P F c 8 e s ( 0 7 _ c0 F Fc s c s ( 3 3 _ c0 c s Ph ( 3 F 5 5 1 T b 9 a 0 5 9 m O u 1 0 R2 3 1 1 N 2 P 0 2 0 2 4 2 0 2 . C GSRETS 2 F SAGGSPSLAQ SEYGW CGG WPG G SFVLSCK TNKGF SYGRGSS GVH e cn A SSTV e r . 1 Y 3 T 2 A SG FGAAPN S R WCYVHWGMH ASSVVNGSPGY GA QQMVSLFAR Y TE e f YPSPEPKNESVGYSSGC e r D SGSSPENKSQATNSD E PGAPCETCNDSR I TGVTCPGGGTFSFRSA . DKTTRKYQGSGGVSKYV e A V RL GGY GYTRMS THPKEFNSLAC VSVI SEGGALSGKPLK . i VSYSSFF SAVSMKNGGDGGVV LVQEH , s S SGSTTENVSSGY GS N ELSSKDKRCGSFSTA ASVS LLTGS DN M YGA LEKS GAGI S GI TG GSDK e ASE c Y n SSANPQEGAS VVTSKWWEPDSI GYQLPPD eu QL GG GGI GSHP YKPVLQS TWSGA QACD CV SE L Y LPDVS A I ATGKPQI A LI YG SL LGE LGT WGI FA FK q e YV RGS EEYRCCYL WGSPHDLKGCKLYGKVS L sl . ) T I Y NK L LGI GSVVS GWSGQKDI TTYTR L R L SS E LQS GQ L F LLSSPG FVPKPGGF PAI YSPT L a s r T i n e t S K N PGE GLVDPVVAGN GWVVT LYQQLAGL LAS g CKVYETRFGM KLASVFT i GYGPV D P R KG LKV TQSKAVS r t o e l GL R TWPLDDEPQ NQKSGPQL GNP Y PPFFPS TGVKHKLRSP V I VNQK S GPVPGF TVDATL oi c KRSY t l a T KT G F QVAQKKVV LSAQ L V LVT QA KQGVAF RDHG SVL LRL YCH LQ AYGWGGI KT e CGG FS LSG HPES v i t t i d GN VYGETW LEWGMRKVD l an R QTRVI WEPKSSI SGKS WASPSSHSFADQKTK e r a G YG K WCW DI SGST TI N LSN CDT LA YETKDPTED MK PNYVMLANSRQGSD s l d RACQQVQNTD AYQQ no A SG W VF S AMTH ASSI ESSFV PD LQDTLR DAGPENSNSS LCWGE i o bn QI S YCSPEKCSSSSFYALFT t a i A WGYGVDAVTGVT G Y SGSRV Y I SYNA PPMSEF SYNCRT SYWTHPKMFYSF EVV GTYVVS i cn SSFSADNE i f wo T VLPI QGSR FVSVTTENE FS STLCFVAKTTDSGG GASI N I DQ d K SATTKKRGSGADESDTS oh s GS Q GV SSI DYCNKSGG AS A FDVDASQSACAQI I N m DGVTVSYG ( e GSFG GSAPCNPQQCS L VK RSYG nr a SII V L QTS CSVA ASEVN PKNGRGW SSSA AAE KTVPSH FKPVP ELWC TRI TYYR LLSAQGS o QQ L i tns e I T I TQCDETCEGSPGY CSYEVCSTTSNCQA eu GG TC Y EQ VYS LARK ISYP EQGVG GGYTST L S LDKVKGVKVA DQD FGMI TCN vi d 6 9 LG RY VRDGQ TR L R L MKGGSGFSS L K L V LKVP DEPLTY YVYDV i n t s e r CW DASSLGN KL GFGNT SG EGASGVT KLGGKESVCDVYET LGPQ WR VRTAK nR PKVSQV LTDFWC e sD GY V PF ) SDEGMDN L PQKPWWGDGVV PSGSI I LS TPQG NLK ATN SAL NFQY L SGNG KVD SEQE VG e r C p; AADAP QGB- S L QQLYKDE PQVSA LDGSI GGK QV LKSATSKG PVKFS LI SGSAPKR AFSTGSN QAN e d en RS 2 VS BSL V DRSSLV PI SGTPH GNF LLI ADS VSLI Y SLGY LGGE LGGSGKE SSL DS LLEF h t WPDKGSHPKGPT NDI SDSFE VLR ISSSS ISRS f l i PK or e WGG MGI LL ST KFTPKKPGCLG TVETSQDTQTPTYT sdn AHr QLG TTSS ENVLYQP KYS DSVFTQG LGPFEI STS VKNHSK IED LT MGSVFSLFV l e ERQTNP p ud NRo SD f MF n DTVVKPSGY GVKHGV PTAAGQ L QRSI CVPV GSYDAPI Q SGTFNS DGSGKMDTL L S L m an a SR Ai a QV m DG SQI SGVV QT FL LR YAN CH LKLQS QK TR VS DV QKPT AA V LQ TVP VLP AT TS GG SGA G WI Q DG SC VG x Q e el d ho ody t b - nn i m -- - a idn mu l e4 d o 0 - hH - o b n ( o 4 0 p p e swo n h k H - a k( ush G i t _ 3 1 0 0 4 c-9 _ ) 1 0 4 i e s e A w s- 1 _ 2 c s- T r 6-9 2 41 _ c s- dob a, t s 5 5 4 0 2 ) 2 01K2 ) 2 0 i t n ict Cu 0 5 BS 4 G, S 4 Gk n a e s e O -6 G - 3 T 1 -3 C _ 2 e r P p 7 a RP 2 - ( 2 0_ c0 0- G ( 2 30 1 _ c0 0- ) p BR pf i7 r 8 e h ( 6 7 Fc s 3 S Fc s 3 3 a c I L , sP5 1 T 5 L b 0 5 : e c 9 a 9 O 5 n e R4 5 6 l eu1 1 m u N P 0 2 0 2 0 2 b aq Te s 4 2 0 2 SVSKL G GSA SVLSLGS GSA SSS LI E. 2 ATYPWWEP S YV ATYTNEKS YV AVTSM LK. 1 YVLKD3 VS L GK LK YVLDLWGGK LK YVYTG 2 YTQGPPQ NTSS HI A SFQH YQGKWELSFQH YLLDN L Q FL S L REK TTSPDVS LREK TTGKW GG LVDSS SDE NGGSKPQI A W Q LFVT SSDE NGSSPKD AGVSLPK PS VYQ VPY QQ PD LPH A W GWAPSFTY GI FK AG WVAF LLDLPS VSSVPY GQ PD FTPYKGI FA WGQ LPPQ K AWLVF L H L VLNPFGVGH SFS GVLNPVLFQTSFS GNAFV L MFTGVK GHLRV LN E H LVL L TST L MFF GTSVGYEVL V HPSVKHLTST L LFPFVT L MGTSV CSVLEYTCL SPAT CSVGVV LNSPAT CSHPS IATGPST LA AAS I ATGG LRCHAAS I ATVGV WE EPS LDTAN LPYSE VH KVS M I YTL ES S WP FY DT LETT L LAKV EI YTS L E S WPGG DSLV FRY GGACQQV L KGGPPENS LRY E L APS KI KT GG NSHL GGAFVMLRY E KI KT L GGT GLAEPTS GLS PTNCE TRKC PES KLSP Q PT AWWHKLS PV EFV KKDK GPTNCQ PENVPES KLGA KSPVKDK GTSPN CFQ GTS AWWPETCSKTS PWNPE QTSVTTDN PGD QTSCRPMFGPGD AQTYWPE RYDTKDKRG KQQ RYDVTPKEVKQQ RDSVCR V WRVCANSPQ QGE VRVPTEQNQGE VRTPPPK CAP RESH MAPKPVP LQ QF W YVT YV WAPG S CREYKASGQFVTV WAVGT PKNPEQYYS CRPEYK YYCF FYEE VVTYR WGE MAFSHP GVS ALYQ YCFYEVL WGE MA TWA CPKA LYQ YFFSEN NYDK NTKKDQK TA VPDV NNS Y SGN NYDVREYRSNNS YYYVHV G NTKKVGVKSGNG NNVDKRE FTV LDYE SDCVKWR FEGTNPT LK I GSS TA SRS FTV LD DCVDQD VP S QSQ L SEGKYEVI GSS TA KV SRS FTDV LDG RT LQSQ L SECVD GALANFQG SGA FGALTNWPRSGA FAG LKTV SR LASKVKY L AAN SRAASNQSAAN GSRANY ASTPEAFF QTDV ALSTPFGYQTDV ALSASW ACNGK SLMGHPKS CS NDI SG I TQK ANSKQKLFCS RL Q CWC CMEHVAFI TQK ANTPNF WC CMSKQ 7 LSTVET NHKD T S VYYQE S VG L QS LTDE SVPKSTC DI SGVY E YQVG S L QEH LS V 9 TDE SYS S GVK I E TSI C SYVPD L RDTAK TDAV GFAR R EN L F SY GVRN TSCETDRTKR R CT KSDAAN LY SVSVP YQSI EDGFEF GTRND SCTE GN PKP AQV TVVP LP AP VDRS GNPTVSLVDRS T S GNCYQ AEPK PKAKDSVPSAEPK PKPTS QSNL GCKT PYTV QSNLTVP LP PYTV QSAKV VDPLTNK S L Q LFP V PGVGTS L Q LFP VNLTD RKFVSVSY SSNS RDKFVLVKTSSNS LDPGV G e GI SSSEVN S PSPKN PI SNL L S L GGI SSS PSC c GT STNKS SYPI SNL L S L GKF GI SVL SSVVC ne SFGPTCE ST ERKV TTRKP QLTCVGQ GT SFG VI SYQ TFVH ERKPV TVEVNS TPKNQT L CETTC FVG VQ GT SFPS H ERGS KPT VVE u VGNSVMEG MDST VGSVTRKPMDST VGTTP qe LAASLKN QTAV LNAAV S Q Q WS SS LT DG NS E KV DG ST GE C Q Q WS SS SI SYQ ME KGQ NVT DGA STV GE LN C QAS Q WAV SVT SR S H V L H L H _ V_ V_ V V 1 1 _ _ G 4 4 4 I g G _ I 2 g G _ I 2 g G _ I 2 g G _ I g _ GA n 0 c 0 0 2 s o s - c s- c 0 7 0 s - c s- c s ic t i 1 t u i p 1 1 E 1 1 1 1 - 1 1 - E- E- E 1 - E P p r 7 a 7 r s 8 e - 8) 8 8 8 -Ce c 1 x 1 8 2 - 8) x 1 2 - 8) x 1- 8) 8 x 1- 8) x e D 6 ( 6 ( 6 2 ( 6 2 ( 6 2 ( Ph 5 1 T b 9 a D 3 I 0 5 0 6 3 6 1 7 9 m Q: 3 O 3 4 u EOR 7 0 8 O O 0 R9 0 0 R1 1 1 N S N P 2 2 P 2 1 2 P 1 2 4 2 0 2 . G G2 N SSA. 1 SE GSK LYV GSNHKSTFQ K . SKI CS AS VI ED LMDTSWSSSGSDASPQSGAC GSVGSGAYSN QATS 2 SY P TDAVQGGAKGGQSHP LWQSL 3 WKFQ EG LREH DK e WPQVPPVSGYV E . i SATVLAPDGGYTLSSTK TFGNPV EET VCRCQR VSI TCLS V IAS LSPS VPY , D s AL e YYPFG LVCTKT NTSSS KG NGRGGSGQKDTVYYG GGGGQSPGGSSSDPVRTG DS LGPA c . TVSSVSYGSFGWGVSGVKVYET LDAP SSSI FFK n e ) s SSSEPVNGRGAWGGGV RPKGFDQ PYKE FQLTVS SL ur T q ee t t SAPPTKCNESSGGLSG SGVRKPGPSVNAGI Y LW SV STN EV L FQGYSPG GT KYGPL s e l KTTVI SYQGVGLFI LGI AGKKLAQG VLHNGAT KAS l a VS i n i cl I GS a YAV E SM LKGGG NGSGMG EG SGS QVAFFS LLG GCTGGWGEKSSSS C TL H LI Y ALTL RS gi r t i I GSSTGSGLSI AERGE LWPI SGSI SE d WSLSDN LEKTGWPI YGGLDTDPT L V L SEL VHKPI KY ETS oon a E LVT K WGG QMPVD t l d GV L Y LP G GKWEPG PD GE LDGL LGKD KG QVS LKGCKGPLGTE AHKSS SI ED VPGVPLQT TFQQ VMD NVKKK e vo K TS GTGSP I AS I YGGLTPTAGDAPQGG KSGTS i t b PQSF L H LDL L LLPWKVQRYVP LPVSG TCPGD l an AGQ LFVSPSKKVATGKVDVVATTDGG MSKQQE e r in Q VVT LYQPTVQYPKTGWVCKKSSG EFVQGFT sw RI WASVFTPKKRDQTI YGTNSYGGS ENQYVV no VI NPFPGSGYGTVRQGYRVVNGGSFG QSG YS PEW E i ot h WYT VKVHPGWAYFWYAEPKNGRGG PLQAG LYQ a s TTSHG NS i c e r MGVLV LRYANKCCR CH LQG AWA FMCANDCTCEPSS PSFRKQGPGS TWN YRSGN i f VS SG d a A os e YSRGET ASPASS LAQVYYFL NYVI SYEGGVG SEYYNYYTI GTFA KNGGG QKI GRS mu NCT LPNYVHWGNVA TSFSM LGSGSEG PDSSQ L ( i d LFACQQMVAYTSI GSSGDTNEKLG EVQS RPT LSAGA n AN oi TD t s e n r I DY R ) GVGP AS ENSLNFSDQDSEDKLWGTS DSTNPCEKCNSG TRTSI SFESSAAPWEPGG RGAAYFAR LKDVS LGG QRSQSV e vD BVDWHPMFGSSSR LQNCSPQI ASKG G CQK i C -1 ACESVTKEVQSASCSS LNPH LDLI YG KY ALI T FTCYWC n QE t ; nd L ATKTEN GAN DS L R LVDKARGSAACMI TTQ M CR L F Q LFVSPSKL L G L 8 9 KFV ISSRYVG e e P RSPCNCQ TS L VYSLVT LYFQKPVT TGDTAKARN s e KDG r nil r VFEF pe s L d a SNESHPQQS Q GMPFKPVP LWCQR LLRY YDTGYSV GV GTYPV TPSKHKK EETRI TTCSVGAFPNGVVVNGT IESSDERSK en 5 4GQ LYVVYSVYGTVDQKGRL PG SD LAPPT h t u 2 PYDKGVKRYGNSEVSL LYCH LKC SVS YFV f d 4 OQVK VKDQDDTAPKSAPLNETWAQG PLPL Q PSLNP on SSNS s a RV LTLDVP GN EVGFQNS L KCVYRT LVDV LDSQ LGGDKPASNLSEQF VHYV GTPI SLS N L l e l d PdGSGKWPKSE KSSGI SPY MWY KTKS SY T L po n GNLTNQSAPGI SPI SSETCQQVAGY VKNQLCG TTFVQ m a bne c n a SEDANF KATSPKGKV L S FLSQ LGGTS FQT L V LFRPPEN EKSL TCSNN CNMDVSH x e i e m r V GKVAFSSSRTTFQGCR FSG KEPQVTAT TVE enu hwq e a VI S QTFGHEPK I SGPI SE ST L V LGM NI QDTQQTPM SAHKEEVNI GS SR Y Q DG S G C t oh S b a Q RS TN VD ES T D Q T Q A D G G TT S n F W K K R G Q S L i V d s e r eh _4 e su a, t tn G i -- -8 1 I g sn b _ i e s 7 4 o 1 n 1 E- -8 7 ede r D k _8 8 1 3 6 _ G 0 c ob p As s- i tnf C i h n i t 1 c-8 0 s- 6 _ c 2 ) s- S 4 i c 1 t 1 a , so u E-8 e c e c t i wt n a 3 0 2 ) 4G ( e i pr mB-0 S 4 0B- _ c7 F0 cCe 1 n euc ro G ( 7 6-e s-P p - 7 a r 8 6 ) x 2 r e f q e s e F 2 ( - 7_ c0 c 2- 0l o 1 1 7 ( 8 e Ph e D 3 F s 7 H E R s 2 8 5 1 T b : e c a 6nD e I : 6 4 9 9 m 2 l e b r ef QEOOR3 4 1 1u N 1 2 a Te r S N P 1 2 1 2 4 2 0 2 N GVH SGSDASQEGAS GSNHKSQDTV LR GVH. 2 M SASAT GGASNPQSGAD SKI CSI EDTGQI S SST. 1 Q 3 L STVE SGYSHP LWSACE ASYVDPLMSVT ASATVE 2 Y FGC GSQTKVTRQCS V RSA SFGNPECSCYYLA RL WSTVAV SPQ PI QGEF TVP LPDSSR FRGSCA T SK LY RGGESVVKI TTTR LS AA L VATSGGGK SK LY N PVQVK SGSSGDQPDVASN YPG LCKTKGSGA PVQV KS GEH P N SDK VGSVVEVRFGM Y GSGKYRT LDDEGQ L TF SVST SSVNSYGFRGW GEKH SEVNGSGSS SDK D ELSE SGGG I V LWPKSGPPQY SPPPKNGSPGA ESE K YPPYD AGYSTNQ VSQVV AGVTRCEGVGYY LYPPY IST G F GI FA I LG GFKGKY LALLT SKT KYPQGGGT GI DA R RFVK S GGSI AQ GWGVEAF KFSS LS ISGN GK I GT S YSV AVI SE SM LKGGSGFSGFFK NGALSGSRVS GN I YSL RGE LPT L I YGLWP GLDI STGST DSPLG TSN I GS ED WSS LTG DNSEGKDG SGGV VKI YSL LPT L A LAAT L W LGKDTEKSSF R VTSKLWGAGSI I V LLAT SKVS SSP KGGAHSI EDQDTV LI S E L Y LPWEPDG TKAVS AVPTS YKKRL GI KS PLP Y PVAGVPLT QT GV LGKDV KTVQGDAVMG VPPI QSVEFR KGTGSP S I YSI GGPTS SPQ HI A L DS L L LGY GEQP LGKRL GI KSY YTV LP TKET VS GPKRY TVDVLPT VVAKTDG SSSG PQ GGK AGQ L F LLSSKPGCWPET VFV VTPKPGKLKVS NGQKD KGWYCNKGSGA QRI WASLYFQQG LGYFQKD G WQQTK AGYGS QTI D QGYGTVSYGFGW V PPV FWREVNNGRGSS GTYGVPAQTKS WI NFGSVKHPTAGYGD GWWQ VLSGQ YAYAPK FE WNDCFTCEGS KPSPVGAY TYTTGVVNKL GY MSHL LNVAT ALPSYRYQGGGTSAGVGLRA EYCH LQKQSWQQ QTRVSSGFE YG DVAT MFWGV YI GNVI SEGGSGFSYRSPTSS LAE CWGWGV CG IASSGSE TTFSA TI SYQ I GTFSM LKNGASGVSA TGSGLGGKNCT LAPNYSVHW MSGGGSGS LFMHI SYEQ WEST ELPQSS DSSN SSGD QDSEDKN LEKD WGSGV GSI I V LLFACQQ TI DYVGPENVSQAARST NTN S SDQSN CGADG SSAAPWEPA KGQTS AY SQ QFAR NCLSKDV PQI AS LDG SI YSI G GYGGASPEKCNDSRSSG QSTNCTRTSSS GVDWHPMFYSLAADS SSCFQTS GLCQL CSS LNPH LDL L LGE L AESTKEVVSFYCQQ L PTI T AWTCA YN I TTQ M CR L F L SSK FVPKPGCWCAV GKLS L R LTKTENSFGTI T VDKARGEGSATCA YN 9 9 QTVYD VYSQ LVT LYFQP GYF SPCNCQSSGASVYD RYRDTV RYGYSVGTYQG LPAR LNESHPQ AACDRTV VDGAK DTGVTPSKHGVAGSMPKVP LWQCC EADGAK WR CAVFWCE GAFPNGVVNPT LQSG FPETR YS L VFWCE MRSDQ ) V Q AE YA PVGDSD YCS L VKGVL K SLRCH LQKRSGQ TVDPLYE YDVVYSI TY KGVKTTR L R LSSDQ AEVG QKRB NF LAN-2 AE SPL GNLY DET AQ SW LEYGWGGQVK GVTV LKDQDVASN PKR DVPVRFG S L Q LAN NYSSSEF RS BL Q TV FAP KRSLG SGKPAPNSH YVMWC SGMAHLN RGKCVYE KWRT LDGDEGM PQ LSSSEF RSK SI S SSQTP LYT FVI L S L PI SSI S ETCQQNVSLF GSG QANDGNLAT PKVPQ NNQSSQ L YPI SP VSTYTV FDTT GE FNPS r S oQT L V LFRPPEEKTC TSR SNDSASEDKATSF PKGKV LA V FSSL GTQLF TTFNPS SAM ARQDN TL L S L f TT n MFQGCR DQQTPMEFNSL VSSSC FFVVI SGKV HEAKFL SSI S TGNKSMDN TL L S L AL SV DG SC VGi Qa I Q DT GSAHK G WT KT KE RNYS Y GV SS FG STQTFGSNPDI SGSPLTGSNQV DGCG mo A Q R T V E T D S F E D S V Q dy m-- k m 4 -1 0 C u b o0 -9 k _3 d n h k H- _8 1 0 1 _ C_ G 0 i t - 9 3 2 ) 3 c 1 c 0 c A SC 0 c s s- w ic 4 t 0 2 8 s- 1 _ s - 4 B- 6 4 3 0 2 ) 4 B 0 G ( H s- - S 4 B- _ c _ 2 4 0 F0 c BCu e 6 2 OR0 G ( 2 6-e s- -6 P p - P 2- _ c0 2- l o 4 0 2-7 a 7 r 8 e 0 7 ( 7 3 Fc s 0 7 H H 0 7 Ph 5 1 T 3 8 b 9 a 6 4 9 m u 5 O 1 R6 1 7 1 8 1 1 N 2 P 2 2 1 2 4 2 0 2 . 2 KYGSGY P GGSQT GKFG VLSCKT TNSKGSGG GAV SGADASQSGCN YGASGS STE GGYSSNPQAASM . 1 NGSFGN ASSVVNGFGA FGSCA SGQKHP LWQTSL Q 3 2 NESRGG WPSE PGSGSSSSGPPKNGRGW TCESSSS R A SPK LY GST V S NPVTRCQR LLY FGGEEVCSI TCSV QGPGSVAKTVTRKPGPVGY VQK RGSSSGVQKTYGT GGVSGKYSVI SYEQGGGY GEDHK SGSVDPDVVRYGN NGG SGSGGV LYTAV KGNGSGT SSE PGSGTVETDTAPKS EKAGI Y WGI LGSTFSSM LGSGEG GGS SSS A VS LTNEGKLGA LPY V GV DPD GGGI Y LYRL TWPKGFQ VDVND EPGG V GSI AQGTSDLW GWGVVY LKPWEGY PGGS SI FA S GI G AFK A I I S LGSGNQSSER GI AQFGY LAPGK GI S A L SR IYGE GLWS LI I LTGKDV SPQI AS LGRGSY DV I S YSL FSWGKKFS PT AGEWVEAKFL Q SLSGT SF DSLS PKL LGGKDI GGSPHDS GTYQ F Y LLVSLI YGI GS SL LGWSL LAL AT RGLLP SS S I YGGDDI STGPI SER VG FQKG TPLVPAE AGLGQ L G GYP W LVF P VT LYKK QPGGE LVKPV GKTEKDSST LLN CKPTS L L L GAHI E QTQAS KHKTVQG V RYKGYASVFTP FP LNG FPSGKYKG LKV GLTQRS KG GI KY PLP VAGSPD LTFQWS QGVAVMDTSSV CH LPK KTVDPATGVVVHGVPGPET QGTWVA L I YQGSHG VL LRYANPTA CH LKV QKVS P KQGQKD KT YD PQ K GVR PKTVDV VVP LP GGAT VATV TDSGYV GGYTQ WAEQGYGRSGETSAQQTR QTGS KGWYGCKNKSSGNT SHYFWRAVDSPSLEYGVI WY VMWAY QD QTI YRTSYGGSGG DCWGT LANSVH CWI NWGQ QGWAVVNGSGWQ QVANSFGGAPYQMWSGTYSFE YFYCEPKNGGFGGAG NSLPNYMHGCPQENVVFMTSLATV WADFTCEPSR GW KCYI GFVARSPEKSWAAGQTS ANPSYRKQGSPGSVL TSTTSA MF SNDRNCRTCSGYRNDE L NFVI SYGGVGLNF EVI GTF ENSSGDS ES LACWSTHPKMS EF GS VI SYNLAN CSSQ YI GSA YSSS ) TTFT E DM LKNSGG GSGMCGS RGQD S SSS YAASFVTTENST FVSN DI GTGETGEKEG A FYTKKRGQSSSI DYSFG BSSSADN L GLGSI ATE PQ PQA LWQF CNCR LGTVDAC SS LSNSAATPECNPQADGVEGS - PSHVP LQQTS CSVASG ASSAQ L 1 QD L SSA DAYAR FCLK W SPW KDEVPT IAS LSGWP AGE LD CG TR CSI TQRMADEFKPETW QCDQCA P QNS LNPQ S LKGKG VKTVCYLSQ LC QDRYGYS LAY RLDE KVVYRI T VSTC E YS KGQKVYLAC RLI TYN TYTD s C V a I TS MP TQCR L H SQ L F LLD I YG FVSSL TPYKLKG LGL PT 1 AW 0 1 PV ET LDT GAGV PTR LSSNV LKDPDRTR L DVYEVDASSVAK 0 NRFWC3 2 VRYYGYVL GVSVFQTPV PTVQT RY RPKVFQN SSD GMCV RTLGFG EVKSG GKWPKVSDEGMDD GEQE 2 VGODT GAPTPSG QN KY KGV HKKVD NGTWR QYAPLNPQ LLTNFQSAPPQ LVPKRRVF SGVV L PGCA GL S L GDQYANKGV QYSQ LAN P SDEV LNL LRCH LKC R KFSQ LGK VATSVKL S L Q L VVASEF dAPGD YT AQGMA AF KSSPSG V LT P I SI SGNGKEAFS PKF SLT SSI GNS LI SRS PKn T a S L Q LGGREPASW NLSEHQFYC YVYF ISGSSETFGKGHDI S PSGKST LYV SSSI SPYVM YNY TDQT L V L GSSNETGST LGSSPTFP m r AP ET QVWGNT KSTT RSNTVHKDQTSNSFNS a SI S FCPQNSAYTFA IED L FQGEDSNSI ESTFED DN b T L V LRPEEKCLNST PAVM IQDTQSQAV LRK PP I SSI CYVDPD LMDVKQ VQTVI STTGL L S CL a Q G F TTFQG VQCTRTSN PMFSGSFD GE LPD AT TSG GSG GGW GSQ AVTS EFPTVA RA LQ TVP VLPV AP TDG SSQ GQTM SF RI QS DGV SVQn i M SH T3 7 I QDTEA SWHTKTEEVNI GSRSSA AR L 0 D G G S K K R G Q S A S 4 - 2 6 1 - 2 / - 1 0 2 b o- 27 k 0 1 _ C_ 2 n k1 O- 1 E- G 0 c 2 ) As s- S 2 . 4 0 9 c W0 c 8 1-8 i c 4G ( s- 4 m or s- 6- t 0 Cu H e - 1 _ c3 F0 0 c H f 3 0 2 ) I S s- -1 7 4B-0 S 4 D P p 7 a 7 r 0- -e 91 l o 4 0 0- D 91 C2 8 e ( - G ( 2 7 0 3-c c s Ph H B 1 F 1 5 1 T b 9 a 7 4 9 m O u 3 2 4 R5 1 1 N 2 2 2 P 2 2 4 2 0 2 QGPVTCEGFGAW SNQ QGPVTCEGFGA ATVI E. 2 FKTTRKPSRSGGL GAC FKTTRKPSR GWL ASVMK. 1 K 3 2 QSV AAVI SYEQG KGGPVSVN S S K GLF FQ LY QSV AVI SYEQGS KGGPG VSVN YASL GLF DSSS YSS LTG DN L NSSM L AVLTGN SDNSG LEGGSGMG RES ASSM LGN GCS DPE SGGGMG TTV K WKEGI AT VSR YSS SHS NAVLT SDN LEGSGCS GTSPW WKEGI AT GVY LKD TTYKWEGPLSWEP GPK TTYKWEGPLSWEP DLTGPQ GV LLP IFTGKDV T PQI AS LSG AGE LD SP G PFW GV LLP IFTGKDV T PQI AS LSG AGE LD A G KGSP SF L H LV PGS ISQSP FH LDSS LKGCG RLTQE PGSP SI YGKL QVP I SQSFH LDSS LK IYGC GKG TQ L I KGQ LF VVT LV GGQ LLFVPKL GPT NST GQ LLFVPSKL GPT RWASPS GWVVT LYQL G LAW NPL GGWVVT LYQL G LAW GYDP V MV WI DAPSVF PSGTK KYPV PTQT RAS VRY I YAL KY MV IDAPSVF PSGTKVQT V KYPPTVRY WMFG GTHGV LR ELAF GTGV GVHGVVHKKVD L LPS WAFTGV VL LRAN YCH LGT PGWR CA KQS E R P A LG GVHGVVHKKVD E VL LRAN YCH LGT PGWR CA LGYVLY GETS R GKVSPAN QYGETS LAKCMA AG LAY QYGETS LAKCMA GST LPY GWSPSNSEQGFYC TGVK GWSPSNSEQGFYC PDACQ PGT LAYVHQYVYF PT LN PGT LAYVHQYVYF AWGPE AV QDAP RSGCPQQM YN ENV NYT LKN AVAPCQQM YNNYT QYSPER VSSPEKSWGTA QTS QDSGPENV CALYFT QGQK RVSSPEKSWGTA R CALYFT VE GNCTP PWHK WRNCRT MSNNSFDE YGGS RNCRTSNNSFD WNDSTT ISG AAWT SHP TKTEFSG EV YDV I GSGA WFP WGWTP RS YVVT I SASHTKM TEFSG EVI GSGE SA MTTVTKK VDAN SRT SAVKDKARN CGSQSAR L SAS YVT NGT ADV E YSRTVKDKARN CGSRAR L WC QSSAS AYPCH YESKV FCPCNPQQSGCN SS TYESHP L AAS LM I G LV SA SG FCPCNPQSGCN N YESHP LQAAS LM SFVP AFPE YEV YYPFKVETWQT Q L NDA TYYPFKVETWQT Q L TTDVG GVYPEVYRCSR LY SDK GVYPEVYRCSR LY FDKKKD SADVGVSKI TQSV SWVP SADVGVSKI TQSV GETV V ATDKKDQDTCYGT SAS ATDKKDQDTCGT SAK LDVY KEV S LKVPVVRYGN GAS KEV LKVPVVY RYGN ALCKW CS DYERT V LDTPK SCD CS DYERT LDTPK CSG LTNF 2 KR LC GV LKW TNPQKSGAQ VFVS RN CYA S D I SYK VR LC GV LKW TNPQKSGAQS VFVND S LN MAN ASKV 0 1 VSANFGVSDEGK TVDW KVSANFG SDRK R L Q LTPKE SS LASKKLAP I RAEAS ) s SS LASKKV LAEPG S GHP SETPVEAFS GST QDVCmSETPVAFS GI ST GY LGND GMGKPKFL Q LGF ETVE r GMGKEPKFL Q LGF GTSVE PYGH I SSSSSER GPSAT a PYGH I SSSSSER PNTNH KASNDETGSPI S G T GP b a KASNDETGSPI S G KKS S KVTT ESTSV KNHKD ICSVI ESS V LN DLQT TLTQQA GR L SSGPA AI SYV KTTV FT F VSSNHKD Kn i ETKI CSVI ESS DLQT TL V L TQN QA V SLSK DSI CYV GDSTDV ASS GESYDPAV FSWSS DE SDPT 3 ASSYDPAV FSWSGRPQTVV QAAQVVP LPMD PQTGSAVTPPA LSSI LVT7 0 GES SDPT QVVP LPMD PQTGSAVTGSI STA LPG LCT VLI T LPTVCAKTVGGYV TDSGY QA S 4QAA TQTSCG2 1 V I T LPTVCAKTVGGYV TDSGYQEFRFVSSVE QT VVFG VLSTVNSKS YGGSGSNTGLVTGV LTEH/ 1 L QTVFG VLSTVNSKS YGGSGT SNTGV L QGKSPSPPT QR GS PS SE PV KN NG GG SGG G WQ GS QS KA KT2 V0 2 V QR GS PS SE PV KN NG GG SGG G WQ GV EV PG KV TR S l O o h- _1 1 1 W4 4 mG ( G ( I SD 6_ E- 8 2 o_ 6 r f 1 _ 6 2 4 62 31 7- _ 3 1L _ 30 c _ 30 c G 0 8 4 6 _ 7 0 D7 0s- 1 7 0s- 1 As 2 1 2 ic / ) 4 1 S 4 2 1 P / d4 n 2 1 1 / E - 4 8 2 1 1 / E- 8 t Cu 2 0 I S 1 2 a 1 21- 1 e 2G ( _ D 0 2 7 40 28 2 1- 6 08 6P p 7 a 7 r OcF2 0 O DO_ 2 8 e W- e c s W C ( W2 ) O_ S W2 ) S Ph 5 1 T 3 4 b 9 a 8 3 9 7 O8 9 1 1 m u 6 2 N 2 2 2 R P 2 2 2 2 4 2 0 2 G . 2 NGGVGLF S . 1 SEGGGMG GNAQ LVV C G P SQKVLP APQTGS TVGGAV VSVSNGGGG YK WSSSEVNGSGW 3 2 WKSGC EGI AST SFQ LSY AEDH SKV ECTKTDSG NKSGGYTV L SAPPPKEGF EGLSWE RES WSPYVSYGSSNT YGVTCKPSRSGA GG QKTRYQGPSV VPTSGEP DPER APDEVNGGSGGG TTSVI SEGGVGL IAS LAGLD VSSH YI FAPKNGFGWQ NAVMKNGGGM DSKGCG S G GPK YFKTRCEPGRGAG GSSLGSGKSGC SL PSI YGK YK G LGL SP TVS PT PF LWQ SSL K TI SYEQSSGGW SSS LT GGPSVL SVSDKN LE WGEG PLTSI A W FQL LVAW R GTKTQT QTVE SP P ASAL ATM SLK TGNGVGLN SGGGMF GTY G GV LLPWE GKDVS LGG GGE L KYP VLHPVRY NS NKKTVD NPT L I GVSS LDKN LEG WKSEGCS I YTSPQI AS WR RAS L YASPWEGLGI AT SGSPH PTSWE I AQQF LLDLKGC VSS PKI YGK LGG CHGPGCA I YA WL KY I GSYKDV LAKC SGGEP WGLFTYQLLPA SEQGFMR L YA LKPS WS QS EVT TSPQI A LGGLD DPHDS LKGCG E LWV LAVSLVFTKV SGYPTVQR VHQVYCF P A L KF LLSSI YGKG GDPFPVKHPKKV QMYYNY AG LA GV LSFVPKL L G LGLT KTTGVV LNGT NV KSWGNT TVY AYTA GT TCLNFST P LLK KTDVT LYQ N G KVPA GAHGRCH KN PGQS AQEPVF W SGTPT KYPVQT PGVL LY LAPGW KCC RY AGYGETW LEQGMY MSNGFDE QTS QGTGGVVVHKKTVD QRDSPSSHQFY EFSSGA QGQ RI WVSLRANG WR VVT LANYVMYVYN EVNI GRSR YGK VI NELYCH LPGCCA YAPCQQVWGN RGSSSAL WGSP WYQEPTS LAKGMR WGGPENSAYT SPQQGASN YFT TTSASSEQFYAC I YRSPEKCSLNFS PLQSACM VVVT MSNPNYVHQYVYF WANCRTFNSGF TWAQTS L YGT AGGCQQMWYNY YDCFWTPKMEV SG CR VSCSQR L Q L N Y SS LEV YRSPENVAGNV SYSHTTENI GRS KI TTCSV I GSG SA A NLCQ FLPEKS ACTRTCLY SNNTFA SS NVV FATVKDKARGSSA SQQSA QDVYYGT NDK YNHPMFSGSFDE STPCNPQSAGAC PEVRTG RPT LDAPNK SDV I DVDTKEV KGFQVS S N SWP GAVKTE GAS SVTKDKARNI GSRG SSA FD GEESHP CGQSAR L SAPFKVLW TRQTS L CS YPECSI QR L QSVDRD SAS ADEWCNPQSGAS AAR LDEVVVKTTCS GYSEGK CCYD ) CAQEHPQAACN CSKKGDQDVYG 3 KLAP AFS L Q LGGI ST I STYA DK DS L R LVG KRVELTWRQTS CSLM N RQ S LMV LKDVPV YET LRYG DTP 0 1 KFSSSSF VAW B- RSANVYVSI TQLL RQCV RKGAFQ ISTGS KDPI SER VG RQEAS 1 LNEFG K DV TCSY TCL SMRSKDQPDVYGV LL T SYG LKW TNPSV FQYSDV EL G IESST QLTL QNA GE PSVE ATDG PTVEVRY P P GQ LYFVYRTDTG APN GVAN K GTA KGLAP TSVKFFS G PD LTFQ SSTRGAb a PY PWPLGFQS PN PEASL Q LG APVMDSWSVGLPV QVNNSNQKVDVN QKSGKPK SSSE LP APQTG VGGAYTVSG AI SYFT F KdV n LTNL L S LF S KG SELD KV LAPGK V LNGSHNDEI SGS TDPI SV L KTTDS SGGGYTQTSPA LDSE GK V a GSCVGQVEAF KFS L Q LG GI S GD T GKTSVHKSST L KNSI ED LQTQ NKGS NGPSI LTDGNVHP SSSSF GI SSI CVPVTFQ SY GSGQQACS B- SDSTDI SGSI SER STSYDAPMDS VNGSG GTSVG7 EKAVETKDPTV LG EFPTVPPQTG KNEGFGW LTLE 4VVI STEH SSL N V LRGAQVLTVGG CK YPG QSRGAWV GS PG SG VL NSG QST KAH KTD V CQTFGSCASVI EPD LQTTFQQASQQLPTVA ba Q R K Y D A V M D S W S Q A FG TDSG LC TK NK YS GG SG S F - hi t 2 ) - S c F- 7 2 6_ wt 4 a G( - b o 0 c nk s- 37 m r 3 _ c 1 1 G 0 4 o f 0 c F - 7 0 8 c 1 c E- As 2 1 / b 8 1 As - l e 4o h s- 1 s - 1 3-8 i ct m0 u 2 0 s s B- _ 1 0 6- 6d eE-8 B- 2 ) Ce P p 2 7 o a 7 r O rC2 - s 07 s o r c 1 - 0 86 2- S 7 4 3 8 e G ( Ph W ( 4 1 5 1 T b 9 a 9 4 9 m O u 0 3 R1 2 1 1 N 2 P 3 2 3 2 4 2 0 2 . 2 Q GTP STE G . 1 G S GAKKRGQSAS GQACRTSNGSF AGKH GGC SGWDACQSGCN SSWTHPMEFSSG 3 W 2 L SG FSN SSA N RTV G Y FGSSSNPQAAT SN SFK LV R K SSA PGYKHP PVLTWQSS LM Q RCQR LL GG SGSATK KTEVI GR RNSSSA Y FGYDKASGQQSA F SKI C RQEH VGYEEVYVSI TCSV RGSYSNPQSGC G P S VSSY S T PDKE GSGTFSSGQKDTVYYGT S N PGI TS DKHVP L AA WQTS L T G SY GGVDP TG VG P T CS E SPQ VPD ASGKVEVRAPKS GGVSEECRSI TQR L P EA LT GSPA LDGVVYRT LDGFQN SGGSVGVKTCS D LPG L AI FK SGSI LWPKVDV LD ASASVDQDVYG G YFS I LFVS L AGI TNQSSEGK LTGVKVPEVRYTG G GV L GI GGFGVAP V TD P T GSS PS GS GPT L D IYSYQKVKALFS L Q LGGI ST GG Y GGCV L R WPL A KGFQ R WT I YGP KV RAT Y L LTTV I YAS L LGE LVTS L KGLGE CW KFSSSF KGI ATNQSV PGKLP YDI SSS TGPI SER TVG I YS LGWG EQFKGKY LSDV L AEG FSPG D KSV LRS PGGFEKDSSLTL QNA LGLGVEAFL Q LG R PASS KPI KY QG LPAHI E QF SKPGGKWPKSSSS A PSS L PETS GVAGSPD LTDQWSAG LDI SGSPI SE R K A G S KV C PI K D PTQSVDAVMTS E F KVY L GK PTK KL S QKRS TVDVVP LPPQVGGSAVKG SGYK YVGLGN VPPFETD AHKSSQT L V L Q Y QKG KGD QGWGVAKTTDSGGGTLPTQDSI ED TKVRSVPLTT VMFQ DS V QTSS QQQ YCGGHCNKGSSNGQKTVYDA PPQTG AS YG QGFET WSGMRTVSYGGSGGQQGWAVVLPVGG D WQQ E SGL YAV LFA A VFV WNS QA AA A TNDEVNGFGWGYTCD SRPKCNEGRGAWWGMKVATTDSG RCKNKSGG LPE NDSATKPSSGGLALFDATSYGGSS RL WSGP GFT YTI GQ NSLCRYQGPSV TS SSS LNYVYCVEVNGGSG FFYI SEGGVGGMFDNGSYPKNGFG S I SYH I GTN YS TM LKNGSGCGI N I DSYTCEPSRG N S M QTSVG SSG QDS VSS EFGSATGSGEG GASDN LEKLGI ASTESDFVRK QTI I SA GSG SI SYEQGPS QL S Y ASS SSYQ L SGADK WGTSWPSYFD KGNGVGG V QDTT L AFA SACEPWEPGGE LDAGGAM LGSGGSG T CSA QNN Q NK I TQG CSD CCYS LA RLKD P TCS PQV IAS LGG S CGQQS LKGKGCQAAT KTDKN LEKE WGLGS 4 S VYN I TTQV CK I TTYTR LSSNF L H LD VSSI YGGLI TCCV WEPTG KL N RYW VYWC VAGMF L G LPTTY SPKDVSGG 0 1 D DTS RYQE RFGQVT LPYFQTKPVAWVYT TQT L S LPQ K GA YRTS L KPH I A LG DS LKGG IS VFV DTVG DD DT GAFKR GEPLSV PQYPSVGKYPVR DA HKKVDRGFDTL ESF LLSSI Y FVPKLG L T SEV F APPCV SSDAN EEF V S ) SQ L V LVTGGVRVANGT PGWCAV RSEPSV PQVT SLY VFQL TKV PT RG S L QE LPFKA PS LPRPKTDA SLS I N S SGN GKL LEYTCH S LAKC QGMYAA KN CS L Q L VKPSGY GVKHPV KK A SS SPI SYDE KVSQ KSLY PSFVB- I SNPS 7 4S DST PLGS TSNP EDAS PNLSE YVHQF MYVY YNF NYS VSSLS PI SGSGV PDL LRV LN T YCH LGPG KC WSSST VQLTV LKDSTN LL L S L Cr SFDV LR ISCPQQ ENVWG SAYTFASST LTGK SI SEPTSW LAEQGF AKTFCVQTTCG QTTQTPEKCLNSDQFETANSHQV Y MD KMFVQo f MGVFCRTSNGFET MDQVPYVMYY YV TLQTG LT DVHn NT GV DG SA AN SI Q DT GS AT Vi a I QSERTPMFSSGA TLRCQQVWG DG SS GG KH TK TE EV NI G SR SS AR LI Q DG SQ VG KP PE EN KS CA LY m N ody m-- -- ml e1 1 b1 1 _ k c u o h E - 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D G2 E SGAKKRG G QSAS WP WDACQSGCN SFG VLSCK TNKGGGGQ GVH GS SYGSFGWG SSTV SAG . 1 A FRGSSSNP2 PQAATSM ASSVVNGRGAW GAE SG3 R LS SSA PGYKH PVLWQSL Q ETYRCQR LL Y Y YPSE I GPVPKNSSGGL ST TCEPGPSVN FRGC FG SA RG N VGYTEVVSI TCSV TKTTRKYQGVGLF SKYV SPS M GGFSSGQKTVYYGT DVSVI SEGGGSGMG PLK VG QL S Y AGGVDPDVRTGNK SAVMKNGEGC I ST VQH GGG V LS T DGGKV GVV LYERPT LDAP GFQSN GSS AS LGSG VS LTDN LEKLGSAE GEDK SG GTGWP SASE ELS N SGSII TWQKSVDEV LD VTSK WPGGE LD LPYD YG K AGI GGNFGVSPGK VCVY LPWEVSGKGCG TPA GG S DSYQKKLAQGI S I A LTGKDQ LI YGKGL GI FK GG N I YGEGVAFFS LLGTF GSPHI AS L GGT GFVS L RG D L LGLWEPKSSSSER WQSPLDSSL LLPAW KS I YS K KGCLDI SGSI S G E LGQ L F LVPKKVTQT I YPT L LG IS PGKYETDPST L V LN G VFT LYQPPVRY L LAT LG TF P R QG LGPF AAHKSQTQA KWLAVSVFT GSI ED LTFQWSGNPPSGYKKVD KAS KPG HGTWR PVTS L PG G G N PVQ KT LRSVP SDA PVMD PQTSSSVPFFGVKNPGCAR ARS KG L GGAKPATHGVV L HKCMA KI KYT GV A QKVDVVLPVSGYVQDVL LRYCLQGFYC GPES PT WS QTWGVATDGGYTLRVSYGETWAQVYF KVD KQV SS YG AV CGGCKNTKSSGSNT ASPSLEHYYNY QKK QKT YK W Y SGMHT FARVSVYGG NG GGGWCDT TV L LANDEPKNGSGWQGMKLANS APYVMWGNVA QTS YG QVALYTFS YGD WC GFRGA DRGCQENSNNSD WQQ SG NT QTSRATCESSGGWLYASPPEKCSGFE AGFET VF GG N WQ NDSLCRK SSSSFI SYPGPSVNGCNCRTS EQGVGLFSYYWTPMFI GSGA LVV WA SRSSR AG YSFY KGGGGMGSVSHKEVNQSAAL YNS SG S DI DE I SG GW VSGT VL SFSAM LGNSGSGCSFAVTTEGSGCN I NDQ SYT LNF EGAST DDN LEKE WGLGI ATS TSWEFTTKK PGDVPDAR NSPQAA QQTS LM S QTI I SN QSSS MG SSGACEKPWEPGGEDSAECSH CSR L Q L SYG AD CS QA AKDVS LGGLGAATPKVP LWRI TQSY AQGS QT IAT CCS L RPQI A KGCGKVFPETSTCGVT CQQ L CS WE EP I TYYR LPHDS LI YGKLCSYEVCKVYYGN QA I TQ 5 LD TVTLS ASNF LLSS FVPKL L G LGPTT AWL S LDKVKGV DQDR VDTAPQK I TCN TC 0 S TVY 1 YD VY CG KG RFGM GL DDGQVT LYQKV TS L K T GEPLSVFTPT YPSGYPKVQ KRYTL V SLKVP VDEVCDVYE KWRT LGVFDV LND R PK DTV RY AK DTA SSEGK GFWC GF PAW) VPQ SQ L V LVTGVK GVVHNGT PGWRS APQG LTNQ KNANFGYA LS LP QG LGI ST VD SEQE V VG SD AE QTY ASGNL LRACH LKCCR KASKVKFSSSF APKR P RVDD WRB- S LI SGK Y A7 ST LGSET SNPSS NLA SEQGMAV HQFYCLSTP GSGKEAFS ER YVYFPDGHPKSPI S DI SGST L V LG S L Q LAN S L Q N SSEF RS SL K SS C 4SPTEDAPYVM YNYGKSNETDQTQA SI SPT PI S MR YADCSF R DV L YCF r QTQI SC Q TPQ PEENVWGN KSAYTVS CLNFAEI ST TSVNH SS VKI CSKS VI EDT LMFQ QDSWSP TGSSVST QLY TFV S FNP QT L S TTF NYo f TGVFC TSNGFDQ LRSYDPVVGGAYKTDN MD NVAni M IQSGESRGTRPMEFS VI GSRGE SAQVGS KPTVAP VP LPDSSGG V GYTLM TI QTL GL S CL G QVT T F S a D S G KH TK T E N S S AR L Q AA LQ T V AT T G S S N G m G D S V Q o ) c D S dy F - - d m-1 8 1 e c - mu l e 1 -8 n l e d o h E - k e 8 6 _ C i l s o h hi t _ 21-8 4 2 ) _ 4 hi _ 3 G s w0 c 6 _ 2 c sS 4 2 c t w0 c A i 5 s- 2 c 5 ) - 2G ( s- 2 2 s- 2 ) t 0 4 5 0 H- S 4 0 G_ - c7 0 5 4 F0 c G 0 - 0 5 B- S 4Cu e P p O 7 a r R1 P 0- G ( 3- 9 1 _ c 7 0 c s 0 - b 3o n s- 1 3 1 0- O 3 R6 P 2- G ( 0_ c 7 8 e Ph ( 7 F 3 K E 3 7 F 5 ( 3 2 5 1 T b 0 5 4 5 9 a 9 O7 8 9 O0 1 1 m u R N P 3 2 3 2 3 2 R P 4 2 4 2 0 2 RET. 2 GPSLE F SHQVYC GQACR . 1 S3 AAPN YYYF SSWTHPMFSS A EVI GRGSR WP SFG LCKK STNYGGGG GSGW 2 WCYVQMVWGNY YN GGSTKENSSAL AV TVA SGAKTKRGQSAS YSSVS PSEVN KNGF SRSGA GG SPQENSANFS FGYDASQSGCN YGPPTCEGPSV APEKCL SNGSD RGYSNPQAAS LM I TKVRKPGVGL YCRTFSSFE SSI TSHP LWQTS Q L DTTVI SYQGGGM YT THPMEVI GRGA PVGDKPVTRCQR LY VSV EGNGSGC STKENSSSR SKTKRGQSAALS GGV SGSEECSI T SVVKTCSV SASM YGT GSSS LK TGSEGE KLG TSI A W ADACQSG SSNPQAATC SN AG M LSGSGQD GAVVDVPVV TRYGN A DTPKS VVLDN VTSKL WWG EPGG GGE L I GS YKHVP LWQSL T TRCQR L Q L GGVK CV LYERLKGA VFQ DV LN C D I V I A L Y LP TGKPDV QI AS LK IYGC GK GPEEYSI TCSY GG WSVVKTYGV KGSI ATWP GNFQSSE GY LAPGK GSPHDS L GI S WQSF LLSSL L G LGPA ESGQDVY LVDPVRTGT I YGWQKKFS APNK L LGE LGVAFLSQ LGT E SF L GGQ LVFVPKKV VT LYQPTVQR CKVYET LDFQS KGGWEKSSSER KWLASVFTPKKV KV L RKGDVN PGKLPI SGPI SVG GNPPSGKYHGTW GTWNPQSVSELD A GNDETDSST LLN PFFGV NPGC PAGFGV LAPGGK KG LPFHK QTQAS PATHGVV L HKCM QQK GVKAFFS L Q L I S G SGST PVPASI ED LTFQWS QD KT VARCLQGY VQDVPVMDSSV RSGAYTWAQFVY R E VI WPKSS ISGSPI SEFR QKRS QTVYDAPQTGAK VY AVG LPVGGYYV L WASE DTPSLEY ANSHWYN GN WI NDTDST L V LG YGWDVATTDSGTT CKLPYVMAYT TYEHKSQTQN TCKVKKSGGSGSNG MRACQQVSLNF MT ASS Y VI EPD LTFQA VMD SWGMRCTNY TSWSALFDAVSNGG G SG Q DAGPEN WG YCSPEKTCNGS SSSFG SG NRDVA LA GPPQVGGSVYVYCEVNG SGAYKDNGSYPKEGFG SRGA G GWL SYNCR SYWTPMFI GRS KEVSSA IDCVLTDGGYV FVATSSGTLTI N I D SDSYTCP SG FVRKQGPSVLNF FVSH SAVTTKTKENQSA RGSGC GYCKKG VTNY GSN T NGSGGGQSI I SAI SYGGVGMG FTVDASQAA YFS EKNGGGCS GDAPCNPQQTS L SVAV ASES PVKNGFG Q EGRGWGA GD QGSAM L SGSG ATGEKELGI ATE SAATEPSKHVP LTWCSR RI TQLS CD SLET ARCKPSSGAW QGPSG CQATDN LWG VLI TQKVKWEPTSWP KVFPE GGE LD CSYEVCVSTC KVYG YG 6 RI SYEGGVG NGGGLNFTCCSP VS VYYT L SKPDI A L SGG G T KGCKG L S LDKVKGDQDR PVDTAP 0 Q 1 RLL M LKSGSGMCGRT LPQDL GGL S L K L V LKVETGF SS GNTG DNEKELTG S SI ATEDAS L K GFTL H SF LLSSI Y VPKL L G LPT T AW ESVCDVY WRLVDV PKSEL G GM P KLWEGP GWPVDEVFTYQKVQT SQG LKTNQSAPG QQ LPWV G VLYKD S LG VPQI A KGGE LDGSES APPQVLFTPT NS VRY PNANFGY L S PVGYHPKVD KKASKKFLSQ LGS GTP NF L H LDS SLI YGCKGLS L Q LKK PSL L G LGTSSV LSGSVK GSGVV LNKGT R V CH LPGW LSTPVAFSSE CCAR GSGKE PDGHPKS ISGPSI S T L V L GKFVYKKVPWSI SPDAARY AKGMA GKSNDETDQTQ GSVT LFQPTATPT LGK TW LEQFYC SI STV KSTFQ SNSVGTYPVQYSTSI SESSHQVYF ETSNHI EDMDS EVD VKPGSVKVHKKRDQFETP NGTVRTD VAPNYVQMYYNY VWGNV VK TA Q LRSSI CS YVDPLQTG AVPVGG QI SGVA QT FA AR YCH SLP AKG QCWCAM G MR AI QTQ DGLR SQ VGC KPQ PEN EKS TCA SLY NNF GSS FD Q E VG ) KPT c Q AA V LQ TVG VLPDSG AT TS GG SG S Fd - e b o 2 7 n ) c ne k S 4 i l G s - G 0 4 c 2 c ( _7 0 hi t 4 2 7 c A 0 si c 1 sGc w L c A s- t 1 2 0 P- s- 1 s 1 5 5 2 0 A L 2 ) s- 1 Cu E e - 8 G-3 A L AE-8 0 5 G_ c O-3FS - 41E bG( - 8 P p 7 a r 1- 86 0 - L 33 _ c 1- 8 R 6 P 0- 33 o n_ G1- 8 7e h F ( K P 6 8P 4 3 5 1 T b 1 5 9 a 9 m 1 OR2 1 1u N 4 2 P 4 2 4 2 0 2 Q GSRETLE FVYC GVH FPTKSVI L FTE . 2 G SAGGSPSSHQYYYF SSTV KATDSSV RGC . 1 3 W 2 L SG FGAAPNYVM GNNYV GAE QL NP V N WCQQVW SAYTA STC YFG LVP LPP DSAY T PVK LV F RSGSPENKCLNFS F G PS RGSA SVSSVCGKT GQK S VGAPETSNGSSFD SKYV TSPSTNK SEHK TE GGYCTR GYTHPMF KEVSRGEA PLK DDGPVSYN FDE NI GSSR VQEH NKVTEPVN RNSPY P S D EGSTTE LSSK RGSSA DKACQQSGAL G CS SDK GTSVTKCE SPD N ASEY PAVRKP VI FAK G YGA G GGSSNPQAAT I SHP LWQSS LM LP Q TPD I YSSI SYQ IFA TSVS L MEKG KF N I YVSS L LT GG RGG W SYK T I YGI GPVTRCQR EE LL G SVYSI TC GFK GTSLTGS L LPT L GVKTYSYV KVS L MVY LD EKQAT Y L D LKGWS GE LVDQD PVVYGT RTGN I YST KVETDAPK L LPAL WL T E LTGKN LWGPAS AS RGSPWELSV QSKDVS LQTS L RS R PGC YRLGFQS KPVS QGQ LPPQI AS LGPI KY A P R K A GG LKV LWPKSVDEV LN ATLS GWV VGTNQ C PTP VSPGDK KRY PAYAF L H LD VSSK TS PKQE LD F KVAGFKGLA QQVKAFFS L Q LGGI S GI KT T PE QDPFV VS RMFTST LYFQTLYKTK GS Y QKRGEKSSS A I SEF K D QKK VAHPVSGYHWQD V QT S YGVI WP CWI NDEI SGS TDPS R ST L V LG QTS WR YGD HYVGVKNG GGG LVV L LYGQ FET D WSGTYHKSQTFQNA WQQ MGSFRYCH LTTYV E VFMTSI ED LTDQWSAGE NRT LETT LAENPS A RL WAAS SGYSGVPVMTS RDAPQGGSSLFVTV YAAPSSHGVEQ GAVYNS NTCGAPNVMNSHS S I SGNAVVG LPTVSGYKDI DE FYSCFQVYSN N SYTLCVATDSG YV LI NDQ TYVNPQNSI VGG M QSI DFCKKGSGSTNTSTS YAWPEEKTCSQS QL S Y ASDGYTNY SV GG AVSNGSFGGGGQ QSI I N GTS AYGS SADVCR S TPPMEFQF VSCYQ LA V QT T CSVSEPVKNEGRGW N QADTCPSSGAGQG KEVPGKENSYN GWCQQQ L CSPYTKQGRVD 7 K I TTCCERKQGPS S VYS LAI SYGGVGVL LNFI TTCA SR YN VLE SPKASECGV FSNPQI SVK 0 1 N RY D DTR L R L ENGG SM LKSGSGM EGCGVYD K SRTV VSS LYEHP EDVVLTWSDWC RAPEAQE K GA IS VFSNTGEKLG DGMDN LWGTSI ATDAKC AMKRKEYSEEVG WEGFDWE GYV T SEG LDVVKGVKR F APPQ LKPKWEPGG PVEQ DVS LGGE LDSPVG PA KR KTCVGQD KDPVPTRAN ISEFS RG S L Q LQ V N SSSLYVPPQI A KG DS L GTFH LSSI Y LGCGA GKGS GLTL Q SLA SEN KS F VAG LTV RS ESANYERT LV ASWPRSP L K LRPK TYT FV A PI SGNKLVPKLLP SI SPK ATTPNQYS LFNP WS SSSQT LGSSF NVST LYFQK TPVAWP PTVQTST LYT GD FV SASEKF HQGKLFPD STNS LS L AV YKTT MFEVDKPVGY YV GSKHKGKRYQTNP QI TSDVA TVDRTFNS V LTVTVEPKFSQG TSL CG TL D NTQTV VVN GV DG SQI SGVAH QT FA AR YC SLP AKG QCW MD CA G MR AI QTL L S L e DG SC VG n Q SN Q ol c VRRCDI SGMGVQ QD KS CT YETD QK ES DI VSVH DG SS AT V 2) 9F S4 s 5 , s i e G( i _ ed 7 4 ) 0 ) d 2 0 2 o 3G o D bi C 0P - 7 4 2 tn – 3 2 3 2 bi t ] 8 3 8 3 n a G c A A L 0 c c a b a 9 _ 9 _ 2B s s- 40 A L s- s 1 - 1 2 0 7 4 mil Si na h Sn i a R ict u B- _ cE- G D U - L C- o r ( 9 c U ( 9 h c I L L-Ce P p 6 7 a r 2- F- 0l 8 7 eo h 1 - 3 86 0- V i t 3 n g a 4Fy v 4Ft i tn 3_ k a mG I 5 u a e G I 5 u h g i a 7 8 e Ph C e c [ g H H g H L e c 5 1 T b n e 7 7 9 a r 7 2 n e r 9 3 4 e f 1 e OR5 6 e f e 1 m u N 4 2 4 2 R P 4 2 4 2 R 4 2 0 2 2 SLTDSV VNAQ SLPTDSV SLLG FGPVSVY. P . 1 K LFG VLV SVP VLP S C K GPT FQ LFG LVVP LP GCQ KKVTEKN 3 2 S LS SVSVGPT SVH QTVPTCNE PSSCKT R NPS DEY PT S PSPSCKT GVST NYSVRKP VNSK PER NGSPTNK SFAV TASSI SYQ YGVEVYN VSSH NYKVVESVYN RTEC TSS L EKG NK TTTPKN GPS KT SSVVT STPKN SG G SA GVTSM LTGNS RCE SPPK NAVVTCE AVKY DVY LDN LE GASSI SKYPQ RF LW GSSR I KYP GLVK NLGK WK AHSS L EG NTQE NSS L SEQG TQEH PT GSPWDEG L I NVTSM Y LK TGN SV S DSP WVTSM LKN H LDK NVQSKPQVS GPT L I WVYTGS SSEY NGQ LPH I A L E I V G LTL GDKN LEK A SL P TLDN LEKNPD GWVF WWG I YAS L A LGGKPWW APA MYALLD FVSS L PS WGSSKDEL L LKPY QSSKDEG LNI FK DPVT LYK ELRG PQVS LKQS WG PQVS LI YF W VS L EMFTSVFQ G Q KW LVP FLHLI A P S DS LATG LA E L A AWQ LVP FLHLI ADS L L LKS LA PT L GYHPSGT VGVKYH GL PDAFVS FP PYFV TST LPS VYKGV FQP L T LY KA K GDAPFVS N PMVFV TST LPSPAT QYGG VYKAAS GI YSLVV FQKVS L PTT LFR EYLN TC TH L QRHPSGTYQQTN PQYHPSGTYGTRS AGAPSLA IRTTVGVKHQGS DVGVKHP VGG LVV LNYGQK I RTGG LVV LNKQI KY QY T RFGANS VI SPFVE QH WSWS WRT LFR EYTC YTAPST L H LW AHGFS WF P SRS II T LFR EYTC EV TH LQE YLS Y KDK WENC RWPQ PEENM KVS YPGANSVHDVT VRAPSLA YYATT GSGANSEWTS I NRSCRTC VH GD YYAVPPMS GLSP SR CFQMG E MAHSPCFQMA LQQ DCTVPKEFV FANPQENVAS LV YANPQNVDGFE TYPGTKEN SCWP GYS EKS VCRTCI GSG TCYWPEEKSN T NNA N LYSCRTCI YT LV FY S TVEYKAQG YAPSNSPE GY YVTPPMS VPKEFSDFK VSSVP STV FATPPMSFDPE GTFYEH VAPGYTKENSQS GTVPKEVE DPGYT SYDQ SDDVVP LQ KENAYS ADKRETW YR ATDEKAQSGGCS SVEKAQSGRYN KSVKDVGVS C TAP LAFSN YEHPET PQCYYDA FTPPFSNPECQG CR LLV CDYEHPQI TQS S DQK 8 S VSCKVPD 0 1 LTVDVVLWI STDAK TSKRETYRVEW T LMNDV KRVELTWTCQ L KR YRVFA VL G LTYEV EANWRT L ESS L V LK CDVVSRDA VGQKQEVS T TC LT TMV LKVVSRYTN S ASNPR CDVGQKDAD AMYTPKFQS PKK L G L LKD TVPDGAVE YEVPQATP QTT LVG LKD TVPDGVV GAS YEVVDK PTESHQG DVKAY LF LSFAN RT LALGA PVAN RT LSTEWC K GQASWNPRGGPV KQASWNPRSPQE KSTVEKF VTSNPI SS ANTP GKSKFQSSI TYT V SEHQG VKY LVA FSLFK LNTP TKSKFQSL Q SDSEV PSEHQG VKY LSLVG ESRCDTG FSSKARN AT GDSTE TCYQKD IES WTS QDTD Q SVEA PKFP SPASI LTS GTS SDTD SVEA PKFPI SEF SST STPTS FRS QMA LKTVSSD L LV ISRN SCDEI STGQ TLAC GV LG EK E KTRN SCDEI STGQTTP FYK FTV V LTVP D FG LV V VP LP QTCTYQKDSVSTH LVCTYQKDSMD P QRVSVGP V EV RP AT KS VI E SD LS EK SA KT V I TT QL RP AT KS VI E SD LI Q DTN GN LS S V QG KS PS SC TKT NT K QE 2 1 BR A 1 S, 1 6 A 2 6 BR 2 A B 1 I 9 0 9 0 I L R 1 2 GA L s L ic – 8 9 2i n 8 a 9 L 2n -] i n n I L 5 L 5 30) 5i n t ] 0 Cu k e c 4 8 h 1 c 0 i 8 a a h e c h c i a h –] 7 1 24 / 1a h c 0y v 41 0ct n 1 Hy v 1 Hct M 0 7 0 2 4 4 y v P p r e 7 a r MG2 I gSa e G2 I h gSg u i L o 4 J G. 9 I g1 J a e 4 G. 9 I 1 J h g G4 i L MGM I GS U ( 1 I a e 7 8 e Ph [ 1 T 4 U H U 3 [ 8 H g 1 [ 3 g N D H 8 5b 7 2 5 2 4 3 9 a 9 m OR7 8 OR9 0 OR1 1 1u N P 4 2 4 2 P 4 2 5 2 P 5 2 4 2 0 2 . SAV 2 F QGPVTCEP ATVI EG SNQ VRSTNVETK. 1 RTEC FKTTRKYQ ASAVM LKNS GAC SSNHI E 3 SG 2 PSA KSVI SE VKY Q G YDSSTGE SFQ AAVSM GLV S LKN TSS LDN LWKGRLS EYS K LK SSI CSP SYVDA SQK YS EH NVS LTGSE TVTSKPWE SDN L KG VPDER PPTVP L GVY LKDI S LPVSSH NA LQTVA ELDK ATT NSE SPY GV L Y LKPWWEG KDVP SDL ATGPQA GSP DSGPSK YNP SFH LSLSSPW TFG LVCK TNS AP FI FDA I FTG PGSPQI A LK SPH LDS LTQ LVP GQ LF KP VT LYFQRF LQ SVSSSVV QTE GSPSEPK IYFK LLVSS L I SQ GGQF LVSPSI KRWVSVGTNVSP YGPTCK WLVFT LYKGYAPPSVKYNPT L I YKVTRY KPT L GVAVSVFQTVDFGVV LHNRAS YTVI SE PAT MI DPPSGKYWMTGRCH I YAL S KY I GAVM LK AAS WAFTGVVVHE LGGHVL LYT LAL LPS WSSTG KV GTS L E LGHGRANGYGEPSW LEKQSA ESVS LDN L PRS GVYVL LYCH LKVSANSVHPAG KI KYT Q GETSS LAGSDT LPYQMTLA L VY GTSK KV L Y LPW KD QES GWSP PGTANSEHPA ACPQENVSGTK GTGPQ QV YKD TK AVLPYV W QMQYGPEKTCP LLN KN PGSSPH L S QDAGCPQENVRESNCRPMSQTS PQQQF LV WYGD RSSSPEKSCVG T F PWHKEVQGQ I RGLFVT L MGGQ V WRNCRT MSWNDSTTERNYGKS WV WAASV FSFET I SGWTHPKEFMTVTKKASGWG YFP SAPPSV NTV AADSVTTEVN T D WCVCNPQVVTT WSFTGGV GW YPSE YRTKKARGAYPSHP LQYVGT YGTHARY YD DEQ S S SAV FCPDCNCPQNYEPKVETWRNS EV YKVAT YESHPQSFVAFPEVCS VNN TYPKVLTWTTY V I S LG H IEGGSEPS SNG Y DVGQKGNSDA SRATANY EQS GVFYPEEVYRFDEKKKDPDVSDK GCLAPQ SQ ACQ L SA ATDVGVSKG DKK STV KDQPDAK LLDVYE CVWRPT LSWVP GYGCPE KSGAS SVYVSPE RYA CYTN K D CEV SSL CDV VYEVAS WRT LC GKTNQSSACSD TANCRP SNLANFKGY LCYA CTWTHK 9 I TTAFVK VKR LSG LKTNPK FQSLMASPVKAFFI STYDK T LDASVTT 0 KK 1 V V AN G R L Q LT EK W SAT A RDWC SS LASKKV LS GKHPI SSVRAEAS LTTVDN DEPQEG SETPVEAFFGY LGNDTGQDVTC EVSPCH GQVKR GMGKPKSGTSTVEHKDSGEVET S EPSKV VSLAN PYGSHNDI SGPNKSNSI E AS DPSAP PS LFPEV SI L SEF K RS KATTVETDKSKI CVPLTR LGA KK LYEG K VSSNHKSV LDSYDAVPGGPV V LSDKVKD FT LP STYT ETK FV ASSI CSI E YV D DPLGDSTVP VGRPQ LPS TVVATA Y SI SFT GF DKE PQV LKV DY PFNP GES A DPTVPPGI SA L CKTPA LSV GNCVW SETNS SAAQVLPSTPG LTNKYPSI LT SEKSG LKN QG L L L S L QI T LTVATEFRFVSVSNQACS i c TATF TS G V LTPG LCKTKV L SSEVNT LSTVG i f Q LDANK IQGC DSV GVQ i c SH T s i f QVVF QRV GSSTN PSV SES PVY G KNQ NVK EVPSP PGPTKCE KV TR SK YPV LE QSGTH c eQ QS KA KT V p VV QI STS GPV KEP ie c e i s T G H D d p b ob i s 1 ,1 i t b1 L / A n a LD 2 / 6 1 / / / / D D _6 4 21 214 _ 6 _ 6 _ 6 _ 6 _ 6 _ 6 _2 P- 7 I 1 1 2) 5 1L P- 7 3 7 3 7 3 7 3 7 3 7 4 6 3 7 3 7 3 7d 4 D1QES G 5 A 5 4 3/ 1 D P- 4D 0 4 0 s 2 4 0 4 1 2 1 2 1 1 i 0 n 4 0 2 4 1 2 1 _ 3 2 i 0 n 4 0 2 4 0 4 n a C A _, 1 2 1 2 1 – 5 1 1 L 1 i n ict 7/ 04 u 0 7 1 4n 2 1i Da 7 4 C D – / I h c C- ] r 1 / 2 1 / a / / 7 0 a / / 1 / n 0 2 1 0 2 h 0 c 1 y 2 1 0 2 0 4 2h 111 i a ] tn /1 c 2 0 2 0 2 0 h c e 7 v 3D 37 P x) a h cCe 4M0 2 t i t e z 22 2 v 2 21y v 2 2 2 t on 3 _ 7 4 4 1 y v P p 7 a r G I gGSQh NU ( E Sg i n L a i fP G I gOOOa W W e G I gOO2 02 a e G I gOOOh g i L I n PD, C ( 8 , 1 a e 7 8 e e Ph c [ W H W W H W W W [ E H 1 n 9 9 1 5 5 T b e 9 a r e 5 3 8 3 9 2 5 f e OR3 4 5 OR6 1 1 m u N 2 R P 5 2 5 2 5 2 P 5 2 4 2 0 2 . 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[0207] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination. All combinations of the embodiments pertaining to the invention are specifically embraced by the present invention and are disclosed herein just as if each and every combination was individually and explicitly disclosed. In addition, all sub- combinations of the various embodiments and elements thereof are also specifically embraced by the present invention and are disclosed herein just as if each and every such sub-combination was individually and explicitly disclosed herein. [0208] The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to fall within the scope of the appended claims. [0209] To the extent possible under the respective patent law, all patents, applications, publications, test methods, literature, and other materials cited herein are hereby incorporated by reference. [0210] The following Examples illustrates the invention described above, but is not, however, intended to limit the scope of the invention in any way. Other test models known as such to the person skilled in the pertinent art can also determine the beneficial effects of the claimed invention. Examples Example 1: Generation and testing of antibodies directed against human PDL1: [0211] The PDL1-BDs disclosed herein have been generated, produced and tested following the methods described in detail in the patent applications WO 2019/072868 and WO 2019/072869. 1.1 Generation, identification, screening and testing [0212] Briefly, rabbits were immunized with recombinant human PDL1 extracellular domain. B cells from these immunized rabbits were then subjected to flow-cytometry-based sorting 119915P877PC 23.12.2024 Numab Therapeutics AG procedure. In this sorting procedure PD1/PDL1 neutralizing high-affinity anti-PDL1 antibodies were identified and isolated through detecting their binding to PD-1 extracellular domain, immobilized on protein G beads. The isolated B cells expressing the neutralizing anti-PDL1 antibodies were cultured, and the cultured supernatants collected. The obtained rabbit monoclonal antibodies in each cell culture supernatant were characterized in a high- throughput ELISA for binding to recombinant human PDL1 and cynomolgus PDL1. In addition, neutralization potential of the PDL1/PD-1 interaction as well as of PDL1/B7-1 interaction was determined by competition ELISA as well as by a cell-based reporter gene assay. In addition, binding affinities of antibodies towards human PDL1 were measured by surface plasmon resonance (SPR). The methods used for direct ELISA, competition ELISA and SPR, as well as the results of the characterization are described in detail in the patent applications WO 2019/072868 and WO 2019/072869. 1.2 Cloning and production of rabbit IgGs [0213] Following the identification of the clones, rabbit IgG antibodies were cloned, expressed and purified for further characterization (hit confirmation), as described in the patent applications WO 2019/072868 and WO 2019/072869. 1.3 Humanization and scFv production [0214] The CDRs of the identified rabbit clones were grafted onto Numab’s proprietary VH/VL frameworks to produce stable scFvs, as described in the patent applications WO 2019/072868 and WO 2019/072869. Different grafting variants were designed and produced. Examples of production details for some of them are summarized in Table 7. 1.4 Pharmacodynamics Characterization of humanized scFvs Affinity to human PDL1 [0215] Affinity of the humanized scFvs to human PDL1 was determined by SPR analysis as described in WO 2019/072869. Results are summarized in Table 8. Neutralization of PDL1/PD-1 interaction by NFAT reporter gene assay [0216] Potency of some PDL1 scFvs to neutralize PDL1-binding to PD-1 was assessed by NFAT cell-based reporter gene assay analogous to the method described in 119915P877PC 23.12.2024 Numab Therapeutics AG WO 2019/072869. Potencies are summarized in Table 9, which shows that the tested anti- PDL1 scFvs can efficiently block ligand binding. Species cross-reactivity (binding to cynomolgus monkey and mouse PDL1 by SPR) [0217] Cross-reactivity to cynomolgus PDL1 was measured in a similar assay as used to measure binding to human PDL1, with the recombinant PDL1 produced by Sino Biological. Table 10 summarizes the affinities obtained for all tested scFvs. All tested scFvs that showed binding to human PDL1 also showed binding to cynomolgus PDL1. Selectivity for PDL1 versus PDL2 by SPR [0218] Humanized scFvs were tested for binding to PDL2 by SPR analysis on a T200 device (Biacore, GE Healthcare). In this experiment, Fc-tagged human PDL2 was captured using the Human Antibody Capture kit from GE HealthCare. After each analyte injection cycle the CM5 sensor chip was regenerated, and new antigen was captured. The scFvs were injected as analyte at a concentration of 180 nM diluted in running buffer for 5 min and dissociation of the protein was allowed to proceed for 12 min. No binding to PDL2 was observed for PRO1392 (33-03-G02-sc18) and PRO1347 (37-20-B03-sc09). 1.5 Biophysical characterization of the humanized anti-PDL1 scFvs [0219] Manufacture of material for stability assessment and storage stability study of exemplary humanized anti-PDL1 scFvs has been performed as described in the patent applications WO 2019/072868 and WO 2019/072869. Results of the storage stability study are summarized in Table 11. Example 2: Generation and testing of antibodies directed against human CD47: Aim of the project [0220] The goal of the project was to generate humanized monoclonal antibody variable domains that specifically bind to and neutralize the biological effect of human CD47. A further goal was to identify anti-CD47 antibody variable domains that are potent and stable enough for incorporation into multispecific antibody formats and have a different range of affinities. r ei t tl ] a/ i L 3 ng . 1 -] E 26 s . 10 E 6 6 0 6 0 E 360 E 3 ±+ . 0 ++ ± EE . 1±+ . 0± Fm [ 7 3 6 2 61 . 2 5 1 1 0 524 4 2 8 4 k a- 1 M [ 6 . 5 E 7 . 4 0 . 6 8 3 . 7 8 0 . 3 3 . 6 E 0 . 4 0 . 4 l d i e yl a] i ng 4 Fm . . . k r 4 [ 4 . 7 0 3 . 0 7 0 1 0 3 . 2 4 6 . 0 0 3 5 7 8 3 o we 1 1 kr . 1 m o L a r 4 F H4 VH 4 3 3 3 3 3 V H V H VH VH VH V H V we D m P a r n a FH 44 4 33 3 3 mu V H VH V H V H VH VH V H V h o t D v I 2 9 4 2 1 9 7 4 3 2 0 3 6 3 5 4 D I in 2 9 4 2 1 9 7 4 3 2 0 3 5 4 F i n c e s e t 3 1 7 3 8 3 3 1 7 3 2 2 0 5 2 4 t 3 1 7 o r ORO O OOOO O O3 8 O 3 3 1 7 3 2 2 2 4 1 P P R P R P R PR P R P R P R P or RR OR ORORO O L P P P P P PR P R P D P-i tn ) G; ) ; a ) ; ) ) 86G8 F 6 G8 G y 8 r l a G8 6G8 G8 G; 8G8 _F G _ 6 F 6 _ F _ p F 6 _ F _ 6 F 66 _ F _ F _ 7 G G G m e GG G GG 67 A6 A 7 6 7 x e 767 7 77 L A 6 f A6 A 6 66 __ _ A_ o __ A_ A_ A VL)K LV LV)K s L L)K LV LV_ LV)K 8( 1 8 V ( c s- 1 86 ( . 1 ( 6 c 20 s- 1 c 4 199984 1 i e i V t 8 ( 8 V ( 864 19 (9. 19 (8 (864 1 20 s- T 2 _ 0 c 0 c 0 c 1 c T _ i n 0Ss- f 1 c s- 1 c s- 1 c s- T _ 0 c 0 c 0 c 1 c 1 c T _ 13 0 s 3 0 s 3 0 s 3 0S f 1 A 2 0 2 0 2 0Ss- s s s s 13 0 3 0 3 0 3 0 3 0S ID e G-3 G-3G- 0 31B- B- B- B- 01 : 8 I D e G- G- G- 01B- B- B- B- B1 - 01 nl o 0- 0- 0- T _0 2- 0 2- 0 2- 0 2- T _ l e n 3 0- 3 0- 3 0- T _0 2- 0 2 0 2 0 20 2T _ C 3 3 3 3 3 3H V7 3 7 3 7 3 7 3H V b a l o T C 3 3 3 3 3 3H V7 - 3 7 - - - 3 7 3 7 3 7 3H V 119915P877PC 23.12.2024 Numab Therapeutics AG Table 9: Potencies of exemplary anti-PDL1 scFvs to inhibit the interaction between human PDL1 and PD-1. PDL1/PD-1 PDL1/PD-1 Clone ID Protein ID EC50 Max. inhibition (rel. [nM] to avelumab) [%] 33-03-G02-sc18 PRO1392 0.97 104.5 33-03-G02-sc18 (VL_A67G_F68G; PRO3891 1.36 94.6 VH_T101S_T146K) 37-20-B03-sc09 PRO1347 0.28 100.1 37-20-B03 sc09.1 PRO2230 0.93 91.8 37-20-B03 sc18 PRO4245 1.81 105.6
Figure imgf000116_0001
Table 10: Affinities of exemplary anti-PDL1 scFvs to cynomolgus PDL1 VH Clone ID Protein ID Frame- ka kd KD work [M-1 s-1] [s-1] [M] 33-03-G02-sc18 PRO1392 VH4 5.2 ±0.2E+06 3.4 ±1.9E-05 6.5 ±3.7E-12 33-03-G02-sc18 (VL_A67G_F68G; PRO3891 VH4 5.9 ±0.5E+06 2.9 ±0.4E-05 3.4 ±2.1E-12 VH_T101S_T146K) 37-20-B03-sc09 PRO1347 VH3 6.88E+06 8.77E-05 1.27E-11 37-20-B03 sc09.1 PRO2230 VH3 7.0 ±1.1E+06 8.4 ±2.1E-05 1.2 ±0.2E-11 37-20-B03 sc18 PRO4245 VH3 4.6 ±0.9E+06 1.4 ±0.2E-04 3.2 ±0.9E-11
Figure imgf000116_0002
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Immunization [0221] Two different immunization protocols were applied to a total of nine rabbits to obtain an optimal immune response against human CD47. Rabbits were immunized with recombinantly produced and purified CD47 extracellular domain (ECD) (Sino Biological, Cat. No.12283-HCCH). Prior to immunization, the quality of recombinant human CD47 was analyzed by the manufacturer 1) for purity by SDS-page and SEC-HPLC, and 2) for biological activity by measuring the ability to bind to SIRPα in a functional ELISA. The first group of six rabbits (protocol 1) received 4 injections of 200 µg each throughout 70 days. The second group of three rabbits (protocol 2) received 5 injections of 200 µg each through a period of 112 days. During the course of the immunization, the strength of the humoral immune response against the antigen was qualitatively assessed by determining the maximal dilution (titer) for the serum of each rabbit that still results in detectable binding of the polyclonal serum antibodies to the antigen. Serum antibody titers against the immobilized antigen (recombinant human CD47 as well as recombinant cynomolgus monkey CD47) were assessed using an enzyme-linked immunosorbent assay (ELISA). All nine rabbits immunized with purified human CD47 ECD showed high titers for the human antigen with EC5 up to 4.6 x 107, as well as for cynomolgus CD47 with EC5 up to 4.0 x 107. Additionally, the titers of neutralizing antibodies to block CD47-binding to SIRPα were determined. The nine rabbits showed neutralizing titers with EC5 up to 4.6 x 106. 2.2. Sorting [0222] Prior to the hit identification procedure, a flow-cytometry-based sorting procedure was performed in the presence R-Phycoerythrin (RPE)-labelled human or cynomolgus monkey CD47 ECD, which allows the specific detection and isolation of CD47-binding B- cells. A total of 1.4 x 108 lymphocytes derived from seven rabbits were analyzed in this sorting campaign. Of these, a total of 7,040 B-cells expressing CD47-specific antibodies (IgG) were isolated and individually cultured as single clones for three to four weeks. 2.3. Hit identification General remarks [0223] For hit identification, direct ELISA screens were performed to assess binding to human CD47. Binding to cynomolgus monkey CD47 was evaluated by direct ELISA as well as CELISA using transiently transfected cells expressing cynomolgus CD47. 119915P877PC 23.12.2024 Numab Therapeutics AG Binding to human CD47 by ELISA [0224] For hit identification, i. e. the identification of B cell clones that produce antibodies binding to CD47, cell culture supernatants of the above 7,040 B cell clones were screened for the presence of antibodies that bind to human CD47 by ELISA. This primary hit identification procedure was performed with non-purified antibodies from the cell culture supernatants, as the scale of the high-throughput culture does not allow for purification of the individual rabbit antibodies. The use of cell supernatants allows for ranking of large numbers of antibodies. The ELISA method used assesses the “quantity” of rabbit IgGs bound to recombinant human CD47, however it does not provide any information regarding the affinity or the concentration of the antibodies. In this assay, supernatants from 1807 B cell clones produced a signal that was above background. Inhibition of SIPRα binding to CD47 in a cellular assay and in a competition ELISA [0225] All 1807 human CD47-binding supernatants were further analyzed for their potential to inhibit the binding of SIPRα to CD47 expressing cells by flow-cytometry and for blockage of the interaction of human CD47 with human SIRPα in a competition ELISA. [0226] For this purpose, a flow-cytometry based assay and a receptor ligand competition- ELISA were developed and adapted for use with B cell supernatant. For the flow-cytometry based assay, inhibition of binding of biotinylated SIPRα-Fc fusion protein to Raji cells expressing CD47 by B-cell supernatants was assessed by flow-cytometry. Since the assays could be performed with B cell supernatant as matrix and were sensitive and precise enough, both assays were used for screening. The inhibitory activity of each B cell supernatant was tested using single well analysis (no dose-responses were performed). Therefore, the extent of inhibition observed is not only dependent on the IgG properties, but also on the concentration of rabbit IgGs in the B cell supernatant. [0227] 905 out of 1807 clones blocked the interaction of human CD47 and human SIRPα by more than 50 % and 811 clones inhibited above 70 % in the flow-cytometry based assay. 1335 clones blocked the interaction of human CD47 and human SIPRα by more than 50 % and 1056 clones inhibited above 70% in the competition ELISA.850 hits show at least 50 % inhibition in both assays. Species cross-reactivity (binding to cynomolgus monkey CD47 by ELISA and CELISA) [0228] All 1,807 hits identified in the primary screening were analyzed for species cross- reactivity to cynomolgus monkey by ELISA and by a cell-based ELISA (CELISA). The ability 119915P877PC 23.12.2024 Numab Therapeutics AG of the antibodies to cross-react with cynomolgus CD47 by CELISA was assessed using transiently transfected CHO cells expressing cynomolgus CD47. [0229] 987 clones (52 %) showed binding to cynomolgus CD47 in the direct ELISA and 1034 clones (57 %) showed binding to cynomolgus monkey CD47 by CELISA, respectively. Overall, many antibodies were identified showing cross-reactivity to cynomolgus monkey species. Binding to red blood cells by flow-cytometry [0230] The ability of the rabbit antibodies to bind to human CD47 expressed on red blood cells (RBCs) was assessed by flow cytometry. Human RBCs were incubated with rIgG positive B cell supernatants at 4 °C for 1 hour, followed by the addition of APC-conjugated anti-rabbit Fc secondary antibody at 4 °C for 30 minutes. [0231] 939 clones (55 %) showed binding to RBC. Consequently, 868 (45 %) antibodies did not show any binding to RBCs. These antibodies might have a beneficial safety profile in vivo. Determination of binding affinity to human CD47 [0232] In a secondary hit identification procedure, information on the binding affinities to human CD47 of the 1053 monoclonal rabbit antibodies that were qualified as positive during the primary screening and were blocking the interaction between SIRPα and CD47 and were cyno CD47 cross-reactive, were determined by surface plasmon resonance (SPR). [0233] For the affinity screening by SPR, an antibody specific for the Fc region of rabbit IgGs was immobilized on a sensor chip (SPR-24 Pro Affinity Sensor, High Capacity Amine, Bruker) using a standard amine-coupling procedure. Rabbit monoclonal antibodies in B cell supernatants were captured by the immobilized anti-rabbit IgG antibody. A minimal IgG concentration in the B cell supernatants is required to allow sufficient capture. After capturing of the monoclonal antibodies, human CD47 ECD (extracellular domain) was injected into the flow cells for 3 min at a concentration of 45 nM, and dissociation of the protein from the IgG captured on the sensor chip was allowed to proceed for 5 min. The assay was run at 37°C with a buffer containing 400 mM NaCl to allow better resolution of the off-rates and therefore improve the ranking of the antibodies. The apparent dissociation (kd) and association (ka) rate constants and the apparent dissociation equilibrium constant (KD) were calculated with the analysis software (Analyzer, Bruker) using the 1:1 Langmuir binding model. 119915P877PC 23.12.2024 Numab Therapeutics AG [0234] Binding affinities to human CD47 could be measured for 656 anti-CD47 antibodies. They showed dissociation constants (KD) ranging from 1.7 x 10-12 M to 5.8 x 10-7 M.27 % of the antibodies analyzed had a KD below 0.5 nM. Competition with CC-90002 for binding to Raji cells by flow-cytometry [0235] The ability of anti-CD47 rIgGs in B cell supernatants to compete with the anti-CD47 antibody from Celgene (clone CC-9002) for binding to CD47 expressed on the cell surface was tested by flow-cytometry. For this, inhibition of binding of saturating amounts of Celgene antibody to Raji cells expressing CD47 by B-cell supernatants was assessed by flow- cytometry. [0236] Out of 1054 clones tested 576 clones (55 %) compete with CC-9002 for CD47- binding and 45% of the clones did not compete for binding. The best non-competing rabbit clones were selected for humanization and scFv production. 2.4 Humanization and scFv production [0237] The CDRs of the identified rabbit clones were grafted onto Numab’s proprietary VH/VL frameworks to produce stable scFvs, as described in the patent application WO 2022/136675. Different grafting variants were designed and produced. 2.5 Manufacture of suitable humanized anti-CD47 binding domains for characterization (scFv format) [0238] The CDRs of the identified rabbit clones were grafted onto Numab’s proprietary VH/VL frameworks to produce stable scFvs, as described in the patent applications WO 2019/072868 and WO 2019/072869. Different grafting variants were designed and produced. From the scFvs that exhibited the desired properties, the best one was selected (PRO3703). Production details of PRO3703 are summarized in Table12. [0239] Briefly, the expression of the scFv was performed in CHO cells using the ExpiCHO Expression System (Thermo Fisher Scientific). Expression was conducted according to manufacturer’s instructions. Proteins were purified from clarified harvest by affinity chromatography (Protein L and/or Protein A). If necessary, variant scFvs were polished by SE-chromatography to a final monomeric content > 95 %. For quality control of the manufactured material, standard analytical methods such as ESI-MS, SE-HPLC, UV280 and SDS-PAGE were applied. 119915P877PC 23.12.2024 Numab Therapeutics AG [0240] The mass of the scFvs has for example been verified by the following ESI-MS standard method. Manufactured scFvs were 5 fold diluted with 1 % TFA. Two μl of sample were injected into an ACQUITY UPLC@ BioResolve-RP-mAb 2.7 μm 2.1x150 mm, 450 Å column (Waters, USA) and desalted using a gradient from 15 % to 85 % buffer B (0.1 % formic acid, 75 % propan-2-ol in acetonitrile) at a flow rate of 200 μl/min at 50°C. The MS analysis was performed on a Synapt G2 mass spectrometer directly coupled to the UPLC station. Mass spectra were acquired in the positive-ion mode by scanning the m/z range from 400 to 5,000 Da with a scan duration of 1 s and an interscan delay of 0.1 s. The data were recorded with the MassLynx 4.2 Software (both Waters, UK). Where possible, the recorded m/z data of single peaks were deconvoluted into mass spectra by applying the maximum entropy algorithm MaxEnt1 (MaxLynx). [0241] The monomeric content of the manufactured scFvs has been determined for example by using the following standard SE-HPLC method. Five µg of the respective scFv, typically present at a concentration of from 0.1 to 10 mg/ml , were injected onto a Shodex KW402.5- 4F column using a Hitachi Chromaster HPLC system at 25°C and a flow rate of 0.35 ml/min. The mobile phase was 50 mM sodium acetate, 250 mM sodium chloride in water at pH 6.0. Elution profile was monitored at 280 nm. Table 12: Manufacture of scFvs Protein ID Based on clone ID Final titer [mg/L] Final purity by SE-HPLC [% monomer] PRO3703 70-26-B04-sc03 66.6 99.5 2.6 Pharmacodynamic characterization of suitable humanized anti-CD47-binding domains (scFv format) Binding potency to human CD47 [0242] Affinity of PRO3703 to human CD47 was determined by SPR analysis using an SPR 24 system (Sierra Sensor - Bruker). Human CD47 was immobilized on a CMD200M SPR sensor prism chip (Xantec). Then PRO3703 was injected at concentrations of 10 nM, 2 nM and 0.4 nM on the sensor chip over spots with immobilized human CD47. An empty spot was used as reference. After each analyte cycle, the surface was regenerated with 3 M MgCl2 solution, thereby allowing the next analyte cycle to have only free ligand available. Binding affinity KD was globally calculated by the Bruker SPR software based on the binding association (kon) and dissociation (koff) rate, by fitting a 1:1 Langmuir model to the curves 119915P877PC 23.12.2024 Numab Therapeutics AG obtained by the cycles with the three different concentrations of the analyte. The results are shown in Table 13. Table 13: Affinity of PRO3703 to human CD47. Protein ID scFv Affinity to human CD47 Based on clone ID ka (M-1 s-1) kd (s-1) KD (M) PRO3703 70-26-B04-sc03 2.44 ±0.21E+06 2.90 ±0.17E-02 1.19 ±0.034E-08 Binding to human and cynomolgus PBMCs [0243] The ability of the anti-CD47 scFvs to bind to specific cells within the subset of PBMCs was investigated. It was found that PRO3703 strongly binds to HLA-DR+ CD14+ monocytes and HLA-DR+ CD14- APCs. No binding to any other of the tested PBMC cell types was observed (see Table 14). Table 14: Binding to human and cynomolgus PBMCs. Human / Cynomolgus Monkey HLA-DR+ CD14+ HLA-DR+ CD14- B cells (CD20+) T cells (CD3+) PRO ID EC50 [nM] EC50 [nM] EC50 [nM] EC50 [nM] PRO3703 5.084 / >40nM 3.350 / >40nM >40nM / >40nM >40nM / >40nM 2.7 Biophysical characterization of anti-CD47-binding domains (scFv format) Storage stability study [0244] PRO3703 was subjected to stability studies, such as a four-week stability study, in which the scFvs were formulated in an aqueous buffer (50 mM phosphate citrate buffer (NaCiP), 150 mM NaCl, pH 6.4) at 10 mg/ml and stored at < ‑80°C, 4°C and 40°C for four weeks. At the minimum, the fractions of monomers and oligomers in the formulation were evaluated by integration of SE-HPLC peak areas after two weeks and at the end of each study. Additional time points were recorded for most of the molecules. Table 15 (monomer content) and Table 16 (protein content) compare day 14 (d14) and endpoint measurements obtained at day 28 (d28) of the study. [0245] The tested anti-CD47 scFv PRO3703 appears to be very stable, exhibiting no measurable loss in the monomer content upon storage for 28 days at ‑80°C and 4°C and 119915P877PC 23.12.2024 Numab Therapeutics AG only minimal loss in monomeric content upon storage for 28 days at 40°C. Likewise, PRO3703 exhibited no protein content loss at any tested storage temperature and time point. Thermal unfolding [0246] Thermal stability of PRO3703 was analyzed by nano dynamic scanning fluorimetry (nDSF) using a NanoTemper, allowing the determination of the onset of unfolding (Tonset), midpoint of unfolding (Tm). Tm and Tonset results of duplicate/triplicate measurements are summarized in Table 17. As can be deduced from Table 17, all molecules exhibit high melting temperatures of at least 59°C, most of the molecules exhibit melting temperatures of >60°C or >70°C. Example 3: Generation and testing of antibodies directed against human LILRB2: Aim of the project [0247] The goal of the project was to generate humanized monoclonal antibody variable domains that specifically bind to and neutralize the biological effect of human LILRB2. A further goal was to identify anti-LILRB2 antibody variable domains that are potent and stable enough for incorporation into multispecific antibody formats. 3.1. Immunization [0248] Three different immunization protocols were applied to a total of 12 rabbits to obtain an optimal immune response against human LILRB2. Rabbits were immunized with recombinantly produced and purified LILRB2 extracellular domain (ECD) (Sino Biological, Cat. No.14132-H08H). Prior to immunization, the quality of recombinant human LILRB2 was analyzed by the manufacturer for purity by SDS-page. The first group of three rabbits (protocol 1) received 4 injections of 200 µg each throughout 33 days. The second group of six rabbits (protocol 2) received 4 injections of 200 µg each throughout 70 days. The third group of three rabbits (protocol 3) received 5 injections of 200 µg each through a period of 108 days. During the course of the immunization, the strength of the humoral immune response against the antigen was qualitatively assessed by determining the maximal dilution (titer) for the serum of each rabbit that still results in detectable binding of the polyclonal serum antibodies to the antigen. 8 2y%) 8 aCC ° 2%) 0 3 f y aCC ° 9 0 . 3 DM ( 4 . 0 o DP ( 4 1 -4 s 1y%) e r aCC u 4 1%) C DM ° ( 0 4 1. 0 t a r e y aC DP ° ( 0 4 . 13 -8 p 2y% m e 8 aC)C 0 t 2 y a%C)C 3 DM ( ° 4 . 0 dn a DP ( ° 4 . 0 4 1 y% aC) / L 4 1 ) DM ( C° 4 0 g . 0 0 1 y a%C f DP ( C° 4 . 80 -8 o 2 y%) n 8 ) aCC° i o 2y%C° . t e DM ( 0 8 - 0. 0 t a r t aC DP ( 0 8 - 6 . 6 sno 4 n T 1 d y%) e c 4 ) n aCC° . n 1%C° . a DM ( 0 8 - a . n o c y aC DP ( 0 8 - a . n Tm 8 a 2 ) t f a y aCC ° . 3 e 8 2 ) on DM ( 0 4 9 9 ga r y aCC ° 0 . 5 0 i ot 4 o a t DP ( 4 1 i n 1 ) s 4 y aCC ° . 5 l e DM ( 0 4 9 9 p 1 ) m yaCC P ° r e ( 0 4 5. t e 8 m a D 9 d s r 8 o f 4 2 y n C) . 5 o 2 ) e 2 1 a C DM ( ° 4 9 9 p y u aCC° 2 i c s DP ( 4 . 9 v s e 4 4 d 1 y aC) l o 1 r e M ( C° . 4 69 t 9 n y aC)C p ° 3. m ] D e t DP ( 4 9 e C ° [ . 8 2 ) no 8 ) To Tm 29 5 y aCC° DM ( 0 8 . - 5 c 2 C 99 i n e t y aC° 0 n 6 a . N o r DP ( 8 - 8 gni ] 4 1 ) 4 ) s C ° y aCC p ° DM ( 0 . 8 - a d 1 . n n at . s y a C C° ny a DP ( 0 . u [ 8 - a . n s t v e sn . 3 F o 4 c T 5 0 e td8 sf ya . 6 no 2 0 y )C ot D 9 9 c ino t a p DP ( 2. 9 n e 3 3 e tu m 0 c n 0 c o r r 3 o 0 c e r n us oi t s- 40 ] L m] oi t s- p 4 p i 0 f fC no s- a pi r B- / g % [ r B o t c - e ° 0 i tp 4 0B e cs 6 2- m [ nt e s 6 e 2- s r 4 i r d c -6 mF e D 0 7 t nt en o D 0 7 uon s e 2- S n 3 I 0 C a D 0 7 D :6 C ° 3 0 n o c : 7 cr e i nmo D 7 1 4 , D 1 3 0 t en o O 3 R O l eC I 7 O 3 O l e D I 7 O 3 o r O p : m : P R P b a ° T0 8 ‑ R P R P b a T R P R P CC P M 119915P877PC 23.12.2024 Numab Therapeutics AG [0249] Serum antibody titers against the immobilized antigen (recombinant human LILRB2) were assessed using an enzyme-linked immunosorbent assay (ELISA). All nine rabbits immunized with purified human LILRB2 showed high titers with EC5 up to 1:1.5 x 107. 3.2. Sorting [0250] Prior to the hit identification procedure, a flow-cytometry-based sorting procedure was performed in the presence of R-Phycoerythrin (RPE)-labelled human or cynomolgus monkey LILRB2 ECD, which allows the specific detection and isolation of high-affinity LILRB2-binding B-cells. A total of 9.05 x 107 lymphocytes derived from nine rabbits were analyzed in this sorting campaign. Of these, a total of 10,560 B-cells expressing LILRB2- specific antibodies (IgG) were isolated and individually cultured as single clones for three to four weeks. 3.3. Hit identification General remarks [0251] For hit identification, direct ELISA screens were performed to assess binding to human LILRB2. Binding to cynomolgus and rhesus monkey LILRB2 was evaluated by CELISA using transiently transfected cells expressing cynomolgus or rhesus LILRB2. Binding to human LILRB2 by ELISA [0252] For hit identification, i. e. the identification of B cell clones that produce antibodies binding to LILRB2, cell culture supernatants of the above 10,560 B cell clones were screened for the presence of antibodies that bind to human LILRB2 ECD by ELISA. This primary hit identification procedure was performed with non-purified antibodies from the cell culture supernatants, as the scale of the high-throughput culture does not allow for purification of the individual rabbit antibodies. The use of cell supernatants allows for ranking of large numbers of antibodies. The ELISA method used assesses the “quantity” of rabbit IgGs bound to recombinant human LILRB2, however it does not provide any information regarding the affinity or the concentration of the antibodies. In this assay, supernatants from 1,423 B cell clones produced a signal that was above background. 119915P877PC 23.12.2024 Numab Therapeutics AG Inhibition of HLA-G and HLA-A02 binding to human LILRB2 expressing cells [0253] All 1,423 human LILRB2-binding supernatants were further analyzed for their potential to inhibit the binding of HLA-G tetramers to LILRB2-expressing cells. For some of the supernatants also the potential to inhibit binding of HLA-A02 tetramers to LILRB2- expressing cells was analyzed. [0254] For this purpose, a flow-cytometry based assay was developed. Inhibition of binding of PE-labelled HLA-G tetramers or HLA-A02 multimers, respectively, to CHO cells stably transfected with human LILRB2 by B-cell supernatants was assessed by flow-cytometry. Since the assays could be performed with B cell supernatant as matrix and were sensitive and precise enough, both assays were used for screening. The blocking activity of each B cell supernatant was tested using single well analysis (no dose-responses were performed). Therefore, the extent of inhibition observed is not only dependent on the IgG properties, but also on the concentration of rabbit IgGs in the B cell supernatant. [0255] 661 out of 1,423 clones blocked the interaction of human LILRB2 and human HLA-G tetramer by more than 30 %.94 out of 931 clones blocked the interaction of human LILRB2 and HLA-A02 by more than 50 %. Species cross-reactivity (binding to cynomolgus monkey and rhesus monkey LILRB2 by CELISA) [0256] All 1,423 hits identified in the primary screening were analyzed for species cross- reactivity to cynomolgus monkey and rhesus monkey LILRB2 by a cell-based ELISA (CELISA). The ability of the antibodies to cross-react with cyno and rhesus LILRB2 was assessed by CELISA using transiently transfected HEK293T cells expressing rhesus or cynomolgus LILRB2 with an N-terminal V5-tag. [0257] 141 (9.9 %) and 51 clones (3.6 %) showed binding to cynomolgus monkey and rhesus monkey LILRB2, respectively. Overall, there were only a few antibodies identified showing cross-reactivity to either of the two monkey species. Specificity assessment and mapping of binding region (binding to human LILRA1 and to human LILRB2/LILRA1 chimeras by CELISA) [0258] All 1,423 hits identified in the primary screening were analyzed for cross-reactivity to LILRA1, the LILR family member with the highest sequence identity to LILRB2, by a cell- based ELISA (CELISA). The ability of the antibodies to selectively bind to LILRB2 versus LILRA1 was performed using full-length LILRA1 transiently transfected HEK293T cells. 119915P877PC 23.12.2024 Numab Therapeutics AG 1,177 clones show binding with an optical density (OD) below 1 and are therefore potentially not binding to LILRA1. [0259] LILRB2 (UniProt: Q8N423) is a single-pass transmembrane protein possessing an extracellular domain of four Ig-like domain and an intracellular domain with 3 ITIM-containing motifs. To map the binding regions of the anti-LILRB2 antibodies, chimeric cDNA constructs were generated where each of the extracellular Ig-like domains of LILRB2 was replaced individually by the corresponding Ig-like domain of LILRA1. The clones potentially not showing binding to LILRA1 were tested for binding to cells transiently transfected with the 4 different LILRB2/LILRA1 chimeras. Domain specificity was used to group the LILRB2 specific antibodies into bins. [0260] 86 antibodies showed binding to the N-terminal Ig-like domain 1 of LILRB2, 80 to Ig- like domain 2, 2 to Ig-like domain 3 and 323 to Ig-like domain 4. For the rest of the antibodies no binding region was identified. Determination of binding affinity to human LILRB2 [0261] In a secondary hit identification procedure, information on the binding affinities to human LILRB2 of the approximately 1,177 monoclonal rabbit antibodies that were qualified as positive during the primary screening and were not binding to LILRA1 were determined by surface plasmon resonance (SPR). [0262] For this affinity screening by SPR, an antibody specific for the Fc region of rabbit IgGs was immobilized on a sensor chip (SPR-24 Pro Affinity Sensor, High Capacity Amine, Bruker) using a standard amine-coupling procedure. Rabbit monoclonal antibodies in B cell supernatants were captured by the immobilized anti-rabbit IgG antibody. After capturing of the monoclonal antibodies, human LILRB2 ECD (extracellular domain) was injected into the flow cells for 3 min at a concentration of 45 nM, and dissociation of the protein from the IgG captured on the sensor chip was allowed to proceed for 5 min. The apparent dissociation (kd) and association (ka) rate constants and the apparent dissociation equilibrium constant (KD) were calculated with the analysis software (Analyzer, Bruker) using the 1:1 Langmuir binding model. [0263] For 817 anti-LILRB2 antibodies binding affinities to human LILRB2 could be measured. They showed dissociation constants (KD) ranging from 4.8 x 10-13 M to 1.2 x 10-7 M.50 % of the antibodies analyzed had a KD below 0.5 nM. 119915P877PC 23.12.2024 Numab Therapeutics AG 3.4 Humanization and scFv production [0264] The CDRs of the identified rabbit clones were grafted onto Numab’s proprietary VH/VL frameworks to produce stable scFvs, as described in the patent applications WO 2022/136675. Different grafting variants were designed and produced. 3.5 Manufacture of suitable humanized anti-LILRB2-binding domains for characterization (scFv format) [0265] Humanized scFvs have been produced analogous to the method described in Example 2.5. Production details of the LILRB2 scFvs are summarized in Table 18. [0266] For additional specific characterization of the 68-18-E11 clone, some IgG variants of this clone have been prepared. Production procedure was analogous to that described below for the IgG-based multispecific anti-CD47xPDL1 and anti-CD47xPDL1xLILRB2 antibodies, with the only difference that for PRO3603, no SEC-polishing step was necessary due to high post-capture monomeric purity. For PRO3603, a simple dialysis against the respective final buffer in a dialysis membrane was performed instead. PRO3635 and PRO4710 on the other hand were polished by SEC, as described below. The manufacturing details of produced anti-LILRB2 IgGs are also summarized in Table 18. Table 18: Manufacture of anti-LILRB2 scFvs and IgGs Based on clone Final purity by SE-HPLC Protein ID Final titer [mg/L] ID [% monomer] PRO3744 68-18-E11-sc07 166.8 99.5 PRO3696 69-01-E02-sc06 655.8 99.4 PRO3697 69-06-G04-sc05 365.6 99.2 PRO3603 (IgG1) 68-18-E11-sc02 108 99.7 (after capture) PRO3635 (IgG4) 68-18-E11-sc02 116 99.7 PRO4710 (IgG4) 68-18-E11-sc07 165 97.8 PRO2518 (Reference) J19.H1 (Jounce) 130 98.9 (after capture) PRO2734 (Reference) Merck 146 96.4 (after capture) PRO3483 (Reference) NGM 77 94.7 119915P877PC 23.12.2024 Numab Therapeutics AG 3.6 Pharmacodynamic characterization of suitable humanized anti-LILRB2-binding domains (scFv format) Binding affinity to LILRB2 [0267] Affinities of the selected anti-LILRB2 scFvs to human and cynomolgus LILRB2 were determined by SPR analysis using an SPR 24 system (Sierra Sensor - Bruker). Human or cynomolgus LILRB2 were immobilized on a CMD200M SPR sensor prism chip (Xantec). Then LILRB2 scFvs were injected at concentrations of 10 nM, 2 nM and 0.4 nM on the sensor chip over spots with immobilized human LILRB2. Empty spots were used as references. After each analyte cycle, the surface was regenerated with 3 M MgCl2 solution, thereby allowing the next analyte cycle to have only free ligand available. Binding affinity KD was globally calculated by the Bruker SPR software based on the binding association (kon) and dissociation (koff) rate, by fitting a 1:1 Langmuir model to the curves obtained by the cycles with the three different concentrations of the analyte. The binding affinities towards human LILRB2 are summarized in Table 19. None of the tested anti-LILRB2 scFvs showed measurable binding to cynomolgus LILRB2. Table 19: Affinity of anti-LILRB2 scFvs to human LILRB2. Protein ID scFv Affinity to human LILRB2 Based on clone ID ka (M-1 s-1) kd (s-1) KD (M) PRO3744 68-18-E11-sc07 7.71E+05 1.14E-04 1.48E-10 PRO3696 69-01-E02-sc06 7.36E+05 1.66E-04 2.26E-10 PRO3697 69-06-G04-sc05 9.74E+05 1.28E-04 1.31E-10 Binding of anti-LILRB2 scFvs to CHO K1 cells expressing human LILRB2 [0268] To test binding of Numab’s anti-LILRB2 scFvs PRO3696 and PRO3744 to CHO K1 cells constitutively expressing human LILRB2, a flow cytometry-based assay was developed. [0269] For cell binding experiments, CHO K1 cells expressing human LILRB2 were seeded in a 96-well plate (50,000 cells per well) and 3-fold serial dilutions of the molecules to test were added at concentrations starting at 100 nM to the plates and incubated for 60 min at 4°C on a shaker. Supernatants were removed and detection reagent (Numab’s anti- framework antibody conjugated to Alexa Fluor 647, in-house) was added for flow cytometry analysis. The plates were measured with the flow cytometer Novocyte with a flow rate of 120 119915P877PC 23.12.2024 Numab Therapeutics AG µl/min and the geometric mean of the fluorescence intensity of single cells was calculated. Obtained MFI values were plotted as function of log-transformed concentrations and EC50 values were calculated by using GraphPad prism software. [0270] Table 20 summarises the cell-binding potency data, presenting the EC50 and maximum binding values for PRO3696 and PRO3744. The data revealed robust binding of the scFvs PRO3744 and PRO3696 to CHO cells that express human LILRB2, with EC50 values in the range of 1 nM. Table 20: Binding of anti-LILRB2 scFvs to CHO K1 cells expressing human LILRB2 PRO ID EC50 [nM] Maximum binding (max. MFI sample) PRO3744 0.881 554’603 PRO3696 0.796 760’006 Binding of anti-LILRB2 scFvs to different human and cynomolgus PBMC cell types [0271] Binding of the anti-LILRB2 scFv PRO3744 to different PBMC cell types from human as well as from cynomolgus monkeys was examined within the specified subset of T cells, B cells, and myeloid cells. In particular, the ability of anti-LILRB2 scFv PRO3744 to target HLA- DR+ CD14+ monocytes and HLA-DR+ CD14- APCs from human and cynomolgus monkey was investigated. The method used to assess the binding to these cell-types was analogous to the method described in the preceding section. [0272] The results of the cell-binding measurements are summarized in Table 21. Besides binding to human HLA-DR+ CD14+ monocytes and human HLA-DR+ CD14- APCs, no binding to any other human PBMC cell types was observed. Also, no significant binding to cynomolgus monkey PBMC cell types was observed. Neutralization of LILRB2 ligand interaction (HLA-G, HLA-A3) [0273] To test the potency of the anti-LILRB2 scFvs to block the binding of known LILRB2 ligands to LILRB2, the binding of HLA-G (complex with human H2AFX, RIIPRHLQL, APC labeled, from Creative Biolabs) and HLA-A3 (HLA-A0301 dextramer, PE labelled, from ImmuDex) to CHO cells expressing human LILRB2 was tested in the presence of the anti LILRB2 scFvs PRO3744, PRO3696 and PRO3697 using flow cytometry. [0274] LILRB2 expressing CHO cells (stably transduced to express human LILRB2 sequence, Q8N423) were seeded in a 96-well plate (50,000 cells per well).4-fold serial dilutions of the molecules to test including benchmark anti-LILRB2 antibody J-19 analogue 119915P877PC 23.12.2024 Numab Therapeutics AG (positive control) were added at concentrations ranging from 160 nM to 0.2 pM to the plates and incubated for 60 min at 4°C on a shaker. Supernatants were removed and APC-labelled HLA-G tetramer (from CreativeBiolabs) or HLA-A0301 dextramer labeled with PE (from ImmuDex) were added to the plates at a final concentration of 100 pM (HLA-A0301 dextramer) or at a final dilution of 1:5000 (HLA-G tetramer). After 30 min incubation at 4°C, cells were washed and 50 µl/well of pre-diluted Cytofix solution (1:10 in ddH20, from BD Bioscience) were added for flow cytometry analysis. The plates were measured with the flow cytometer Novocyte with a flow rate of 120 µl/min and the geometric mean of the PE and APC fluorescence intensity of bound LILRB2 ligand, HLA- G or HLA-A3, to single cells was calculated. Obtained MFI values were plotted as function of log-transformed concentrations and EC50 values were calculated by using GraphPad prism software. [0275] The potency data for inhibition of LILRB2/HLA-G and LILRB2/HLA-A3 are summarized in Table 22. The IC50 as well as maximum inhibition values for the anti-LILRB2 scFvs PRO3744, PRO3696 and PRO3697 were normalized to the control IgG antibody J-19 (PRO2518), which exhibits high affinity binding to human LILRB2. [0276] Although being monovalent, all tested scFvs, namely PRO3744, PRO3696, and PRO3697, demonstrated potent and comprehensive inhibition of the LILRB2/HLA-G interaction with potencies similar to the bivalent IgG antibody J-19 (PRO2518). Notably, PRO3744 also achieved complete neutralization of the HLA-A3/LILRB2 interaction, while PRO3696 exhibited partial inhibition, and PRO3697 did not impede HLA-A3 binding. LILRB2 selectivity assessment [0277] To assess whether the anti-LILRB2 antibodies selectively bind to LILRB2 and not to other members of protein family LILRA and LILRB, a cellular ELISA (CELISA) using transiently transfected CHO cells was developed. [0278] The selectivity assessment was performed as follows. The day before CELISA, CHO cells seeded in a 96-well cell culture plate were transiently transfected with plasmids encoding LILRA1 (identifier: O75019-1), LILRA2 (Q8N149-1), LILRA3 (Q8N6C8-1), LILRA4 (P59901-1), LILRA5 (A6NI73-1), LILRA6 (Q6PI73-1), LILRB1 (Q8NHL6-1), LILRB2 (Q8N423-2), LILRB3 (O75022-1), LILRB4 (Q8NHJ6-1) or LILRB5 (O75023-1). Next day, dilution series of molecules to test ranging from 100 nM to 6.4 pM were added to the plates of transfected CHO cells and incubated for 1 h at 37°C in the incubator. I gi gi g g l sl ) e +F] n n i n i n 0 di di di di c 3M Do 0 nb nb nb nb TC ( e 0 g [ 1 x o No N o No N g
Figure imgf000133_0001
)+IF] i n g g g d i nd i nd i nd
Figure imgf000133_0002
l sl e 0 c 2M0 Do e0 0 i BC ( g [ 1 nb i n i n i n x o b No b b N o No N g +R I i n gi n gi n gi n %) 9 D - -F] d d d d . A4 0 i n i n i n i n x n 1 a i o - J L1Mo e0 0 b b b b i HD Cg [ 1 o o o o m . t i x N N N N l e b o h t . l R i n e r ( . 76 . 9 26 7 5. 9 + 4. 0. 6. 9. 0 5 R I D-+F] 0 0 50 10 72 C I A4 L1Mo e 0 0 0 ±8 ± 2 HD Cg [ 1 x 3 . 0 1 ± . 1 3 4 ± / .0 8 6 0 . 5 9 7 C1 I - ) . J0 l ep 3 3 l e C5 ) R I ( ma s 89 6. 7 6 A + 0 . 0 3 A- N A D L C ( I g lsF] i n gi n gi n gi n H /2 ] n M o iti leM0 0 di n d co e 0 b i n d b i n d b i nb B n [ i bh Tg [ 1 x o No N o No R N I L 0 5 3 L I C 9 7 8 . 0 6 8 n i 2 .0 o n 3 1 ) I gi l s n gi n gi n gi ) l + e 0F] M0 0 di n di n d n nd n . x %n 9 1- c 2Do0 b b i b i b a i o J BC ( e g [ 1 x o No N o No N m . i t o le i bh t . i n l e r ( . 1 . 4 . 5 y R d 9 9 8 9 9 9 +R I o D - -F] 7. 9. . 5 . 3 ib t 0 5 A4 L1Mo e 0 0 2 2 4 6 n 0 a C HD Cg [ 1 ± ±±± k x 5 . 3 6 . 3 4 . 5 5 . 7 r o y I c / n 05 9 1- ) we m e t C I . J a o l e C0 l 5 ep m 4 8 5 0 6 3 6 6+ I . . 9 . 04. 2 f ri-t pn R I ( a s . 0 . 0 1 . 1 R D-+F] 15 8 5 1 1 n A4 1Mo0 0 0 ± . 0 ± . 1 ± . 1 ± a i o . n t ai z G-L HDe Cg [ 1 x 6 1 1 2 7 2 27 3 a hti l a AL w r t d u e H / ] -n e n 2B Mn ecn no k oi n t ] i l 2 R [ B I L 0 5 37 958 039 5 Ca rM Tn [ 6 . 100 2 6 . 100 2 s s or R L I I C L . 0 . 1 . 0 4 D I 4 4 7 4 ) * c : L d d : 2 4 4 6 9 7 9 O 3 7 3 e k e ki n 2 D I 7 R OR Oin l l - l e b O 3 6 R O3 6 R O3 R OR P P R P- x X) * a T P P P P 119915P877PC 23.12.2024 Numab Therapeutics AG [0279] As a positive control to confirm successful expression of proteins of interest, transfected CHO cells were also stained with an antibody directed against V5 tag (R&D Systems, cat. MAB8926), as all LILRA and LILRB constructs were designed to express V5 tag at the N-terminus of the proteins of interest. Supernatants were removed and detection reagent (rabbit anti-human Fc fragment, HRP-conjugated, from Jackson ImmunoResearch, cat.309-035-008) was added to the wells. Plates were incubated for 45 min at 37°C in the incubator. Next, cells were washed and 50 µl of TMB substrate was added to each well. HRP reaction was stopped until sufficient color development was achieved by adding 50 µl of stopping solution (1M HCl) to each well. Plates were read by optical density (OD) measurement at 450 nm and at 690 nm as reference wavelength using BioTek Synergy Neo2 multimode reader. Obtained OD values were plotted as function of log-transformed concentrations and EC50 values were calculated by using GraphPad prism software. [0280] As shown in Figures 1, 2 and 3, PRO4710, the IgG4 variant of scFv PRO3744 (68- 18-E11-sc07) as well as PRO3635, the (IgG variant of scFv PRO3064 (68-18-E11-sc02), showed selective binding to LILRB2 while exhibiting no binding to any one of the tested LILRA family members or to LILRB1, LILRB3, LILRB4 or LILRB5. The scFv PRO3153 (69- 01-E02-sc03), i. e. a precursor of PRO3696 (69-01-E02-sc06), exhibited the same selectivity for LILRB2 as PRO4710, when tested at a concentration of 150 nM (see Figure 4). On the other hand, cross-linked PRO3697 (69-06-G04-sc05) showed, besides strong binding to LILRB2, weak binding to LILRB1, but no binding to any of the tested LILRBA family members or to LILRB3, LILRB4 or LILRB5, as seen for the other clones (see Figure 5). LPS-driven cytokine secretion [0281] To evaluate the effect of the LILRB2-BDs of the invention on the cytokine secretion levels of human monocyte-derived macrophages (HMDM), an LPS-driven cytokine secretion assay was performed as described in the following. [0282] HMDMs were generated by culturing CD14+ cells for 6-8 days in complete medium (RPMI 1640, 10 % heat-inactivated bovine serum, Sigma Aldrich) in the presence of 50 ng/ml human macrophage colony stimulating factor (M-CSF) (Peprotech) at 37°C and 5 % CO2. Medium was exchanged and M-CSF replenished on day 3 of the culture.50000 - 100000 HMDM cells were seeded in a flat-bottom 96-well plate and exposed to 100 ng/mL LPS in the presence of a serial dilution of PRO3744, PRO3696, and PRO3697. After 24 h of incubation at 37°C and 5 % CO2, cell culture supernatants were harvested and human interleukin 1β (IL-1β), human interleukin 6 (IL-6) and tumor necrosis cell factor alpha (TNF-α) levels were quantified using the cytometric bead array method (CBA) from LegendPlex kit provided by Biolegend according to kit instructions (cat no.: 740930). Cytokine 119915P877PC 23.12.2024 Numab Therapeutics AG concentrations were interpolated from a standard curve generated for each cytokine and plotted against control conditions for calculation of EC50 values. Data was acquired on Attune NxT Flow Cytometer (Thermo Fisher Scientific) and NovoCyte Quanteon 4025 Flow Cytometer (Agilent) and analyzed by Biolegend’s LegendPLEX Data Analysis Software. [0283] As shown in Table 23, IL-1β, IL-6 and TNF-α levels increased in the dose-dependent manner, indicating the activation of HMDM cells by PRO3744, PRO3696, and PRO3697 in the presence of LPS. Re-polarization of M2 macrophages [0284] Further, to evaluate the potency of the LILRB2-BDs of the invention to stimulate re- polarization of M2 macrophages to their M1 state, a re-polarization assay has been performed, which follows the level of several surface markers that are indicative for re- polarization, such as CD86 (marker for M1 state) and CD163 and CD206 (markers for M2 state). The re-polarization assay was performed as follows. [0285] Human peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats of healthy donors by density gradient centrifugation (STEMCELL Technologies). CD14+ cells were isolated from PBMCs by positive selection with magnetic beads (Miltenyi Biotech, 130- 050-201). HMDMs were generated by culturing CD14+ cells for 6-8 days in complete medium (RPMI 1640, 10 % heat-inactivated bovine serum, Sigma Aldrich) in the presence of 50 ng/ml human macrophage colony stimulating factor (M-CSF) (Peprotech) at 37°C and 5 % CO2. Medium was exchanged and M-CSF replenished on day 3 of the culture. [0286] HMDM cells were exposed to 25 ng/mL IL-4 in the presence of a serial dilution of PRO3744, PRO3696, and PRO3697 at 37°C and 5 % CO2. After 24 h, flow cytometry assessment for the surface markers CD86, CD163 and CD206 was performed. [0287] HMDMs were harvested from culturing flasks on day 6-8 using Accutase (Biowest), seeded at 50’000 cells per well in 96-well flat bottom plates (TPP) and treated with 25 ng/mL IL-4 (Peprotech, 200-04 – 50 µg) together with a serial dilution of test compounds or isotype antibodies (dose range 100 nM – 0.0006 nM) for 24 h at 37°C and 5 % CO2. After 24 h, cells were stained with Zombie Aqua Fixable Viability Kit (BioLegend) for 30 min at 4°C. Cells were incubated with the Fc receptor block (Human TruStain FcX, BioLegend, 422302) for 10 min at room temperature, to prevent nonspecific binding of the staining antibodies. Staining antibodies (all from BioLegend) were added to the cells for 30 min at 4°C: CD163- APC (cat. no.333610), CD206-BV711 (cat. no.321136), CD86-AF488 (cat. no.305414), CD80-PE (cat. no.305208) and CD14-PerCp-Cy5.5 (cat. no.325622). Following washing step to remove primary antibodies, samples were fixed with a BD CellFix kit (BD, 340181): Cells were incubated in 50 µL of fixing solution for 20-30 min at 4°C in the dark. Samples 119915P877PC 23.12.2024 Numab Therapeutics AG were washed, resuspended in PBS and acquired with Attune NxT Flow Cytometer (Thermo Fisher Scientific). Data was analysed with the FlowJo software (Tree Star). [0288] Figure 6 and Table 23, summarize the results obtained for CD86, CD163 and CD206 levels, i. e. markers associated with M1 (pro-inflammatory) and M2 (anti-inflammatory) macrophage polarization, in the presence of the anti-LILRB2 scFvs PRO3744, PRO3696, and PRO3697. For all anti-LILRB2-BDs, a dose dependent increase in the M1 marker CD86 and dose dependent decrease in the M2 markers CD163 and CD206 could be observed. These results indicate a shift of the macrophages towards M1 state (re-polarization), potentially suggesting an enhanced immune response. Table 23: LPS-driven cytokine secretion and repolarization of M2 macrophages LPS-driven cytokine secretion Re-polarization IL-1β EC50 TNF-α IL-6 CD86 CD163 CD206 [nM] EC50 [nM] EC50 [nM] EC50 [nM] EC50 [nM] EC50 [nM] PRO3744 0.7788 1.1031 0.8035 0.1171 0.1070 0.1361 PRO3696 0.4462 0.8912 0.8036 0.2433 0.3715 0.5867 PRO3697 0.2788 2.2044 0.4500 0.2140 0.1726 0.5117 3.7 Biophysical characterization of suitable anti-LILRB2-binding domains (scFv format) Storage stability study [0289] Humanized scFvs were subjected to stability studies, such as a four-week stability study, in which the scFvs were formulated in an aqueous buffer (50 mM NaCiP, 150 mM NaCl, pH 6.4) at 10 mg/ml and stored at < ‑80°C, 4°C and 40°C for four weeks. At the minimum, the fractions of monomers and oligomers in the formulation were evaluated by integration of SE-HPLC peak areas after two weeks and at the end of each study. Additional time points were recorded for most of the molecules. Table 24 (monomer content) and Table 25 (protein content) compare day 14 (d14) and endpoint measurements obtained on day 28 (d28) of the study. [0290] Most molecules exhibited >5 % monomer loss upon storage for 28 days at 4°C. None of the scFvs exhibited considerable protein content loss at any tested storage temperature and time point. Most pronounced differences in monomeric content between molecules were observed in samples stored for 14/28 days at 40°C. 119915P877PC 23.12.2024 Numab Therapeutics AG Thermal unfolding [0291] Thermal stability of scFvs was analyzed by nano dynamic scanning fluorimetry (nDSF) using a NanoTemper, allowing the determination of the onset of unfolding (Tonset), midpoint of unfolding (Tm). Tm and Tonset results of duplicate/triplicate measurements are summarized in Table 26. As can be deduced from Table 26, all molecules exhibit high melting temperatures of at least 59°C, most of the molecules exhibit melting temperatures of >60°C or >70°C. Example 4: Manufacture and characterization of anti-CD47 x PDL1 bispecific antibodies: 4.1 Manufacture of anti-CD47 x PDL1 bispecific antibodies [0292] Manufacture anti-CD47 x PDL1 bispecific antibodies was performed as follows. Expression of multispecific constructs was performed in ExpiCHO-S cells using the transient ExpiCHO expression system (Gibco). Expression cultures were cultivated in batch using shaking flasks for 8 days at 37°C. The culture supernatants were separated from cells by centrifugation followed by a 0.22 μm sterile filtration. [0293] Target proteins were captured from the clarified culture supernatants by protein A affinity chromatography and polished by a size-exclusion chromatography (SEC) step, except for the reference molecule PRO3851 (Innovent), which was polished by a cation- exchange chromatography (CIEX) step. For some molecules, no post-capture polishing step was necessary. [0294] For the quality control of the manufactured material standard analytical methods such as SE-HPLC, SDS-PAGE, and UV280 were used. Manufacturing details of anti-CD47 x PDL1 bispecific antibodies according to the invention as well as of reference anti-CD47 x PDL1 bispecific antibodies are summarized in Table 27. The reference PRO3609 has an (scFv)2- Fc antibody format, which is a format according to the invention, but comprises an CD47-BD that has an intermediate (higher) binding affinity. 8 2y%) aCC DM ° ( 0 4 4 . 0 5 f 8 . o 2 69. 3 s y a%) CC ° er DP ( 0 4 . 53 - . 54 - . 72 -d 4 ) ut n 1 a y% aCC M ° a 4 ) ( 0 4 . 9 r 0 - 9. 52. 2 e 1 p y a%CC ° 0 . 7 . 1 . 4 / L D P ( 4 1 - 0 - 2 - g 0 8 m D 1f 2 e y% t 8 aC) M ( C° 4 . 01 - 0 d .69. 0 n 2 a y a%C) PC° 4. . 91 . 9 o D n / D ( 4 0 - 0 -i ot 4 L t 1% g 4 a rn y aC)C . 008 0 1 1 y%C)C . 2 e D c M ( ° 4 1 - . 5 . 1 f o a DP ( ° 4 9. 04. 1 1 -no 8 . 2%) n t i 8 ) e s a a C ot 2 C n c yC° t a DM ( 0 8 - . 01 - 0. 19. a 0 r tn y a%C° DP ( 0 8 - 3 o .0 . 81 - . 44 - Td eg 4 a r 1 y%) e c C no 4 n 1%)C a m o t aC° DM ( 0 . 8 - a . n . a . n . a . n c a y aC° 0 . a . . a . . a . Tf s t e a DP ( 8 - n n n o l 8 p 2 ) e 8 nm a y aCC ° . 5 . 8 . 9 g 2 ) C i ot a s DM ( 0 4 8 9 1 9 5 9 a r ot y aC° DP ( 0 . 4 2 0 . 1 0 1 . 1 8 0 1 i n no 4 p 1 ) s u y l e s aCC ° . 8 . 3 . 6 p 4 m 1 ) r e yCC ° . 1 . 5 . 8 t e s DM ( 0 4 9 9 2 9 7 9 m a a DP ( 0 4 0 1 0 1 0 1 dr 7 l o t 8 sn 8 o f 3 1n 2 e t y aC)C n DM ( ° . 4 99 . 9 2 2 . 9 9 o 8 p 2 9 u y aC) e C° 9. . 7 . 6 i c v eo s c 4 s DP ( 4 9 0 1 0 1 l d i o c 1C) . . . t 4 r e n 1 ) p r e . s y a C 9 2 0 m DM ( ° 4 9 9 3 9 8 9 e t y aC P oy a n D ( C° 4 8. . 9 3 0 . 1 6 0 1 m eT ]nd o8 8 o o C ° 2 )C c 8 [ 2 ) . . . C n a m 6 4 7 5 5 1 1 m 2 do y aC° M 0 8 . 99 . 17 . 09 i n e t y aC° 0 9. . 7 0 . 0 1 N T 6 6 g n a tp D ( - 9 9 9 o r DP ( 8 - 9 1 1 n ] tu nr 4 e to f 1 ) yaCC p i s ° d 4 ) 0 . a . a . a n u C ° [ t a . s 1y C C° 0 . . a . s v e sn . 1 . 9 . 2 noC c° 0 DM ( 8 - . n . n . n t a a ny a DP ( 8- . n . n a . n F To 4 6 7 5 3 5 i c 4 e td c s r edn 0 n8 0 y . 9 . 1 . 8 o 2 y a) f C . 5 . 5 ot 756 m a a o D 8 9 8 9 9 9 c ino t DP ( 9. 9 0 1 0 1 n e n 0 c 0 s - c s- 0 c s-nC ° p o 4 m, 70506 e t our 75 m oi 06 e r tp 1 1 4 E 0 G2 0E] L] f C° 0 n c o s - c s- 0 c e 8 4 s - rpf o f n 0 c o s - c s- 0 c s- u s i r a c - s 8 -6 -1m e 1- 0- 0- / g % [o i t 1 1 0 2 0 oC i t 1 1 4 0 2 0 e t n s ‑ r f pi r E-8 G-6 E- e ° 0 pi r E- G- E- m D 8 6 9 6 9 6 m [t nt euo os cs e 1- 0 - 1 0- s ru 4d c 8 6 1 F n s e 1- 0- 0- S t en n o c Ce D 8 6 9 6 9 6 o 8 6 9 6 9 6 D o r e : r 4ut 2 a l er e 47 6 C a D : C D 4 7 9 6 9 6 5 2 ° n 4 , 4 : c i nmo D 4 7 7 9 6 6 9 6 6 2 4 D 4 7 7 9 6 6 9 6 t en o bp I am O 3 R O3 RO3 ROR l eC I ° 3 3 3 Te t P P P P b a0 OR OROROR l e I O 3 O3 O3 Oo r p: m : T8 ‑ P P P P b a R RRR T P P P PCC P M 119915P877PC 23.12.2024 Numab Therapeutics AG Table 27: Manufacture of anti-CD47 x PDL1 bispecific antibodies according to the invention and reference constructs Description Post-captur Post-SEC PRO ID e titer [mg/L monomeric PDL1 binder CD47 binder ]* purity [%] PRO3790 37-20-B03-sc09.1 70-26-B04-sc03 84.3 100.0 PRO3609 70-36-C05-sc02 (Reference) 37-20-B03-sc09.1 (intermediate affinity) 245 98.6 PRO3599 CD4 145 99.8 (Reference) 7xPDL1 (Pfizer) PRO3851 CD47xPDL1 ( 442 97.3 (Reference) Innovent) 4.2 Pharmacodynamic characterization of suitable anti-CD47-binding domains Binding affinity to human CD47 and PDL1 [0295] Affinity of PRO3790 to human CD47 and human PDL1 was determined by SPR analysis as described in Examples 2.6 and 3.6. The results are shown in Table 28. Table 28: Affinity of PRO3790 to human CD47 and human PDL1. Protein ID Target Affinity ka (M-1 s-1) kd (s-1) KD (M) PRO3790 human CD47 6.72E+05 2.35E-02 3.50E-08 human PDL1 5.52E+05 4.19E-05 7.59E-11 Binding to cancer cell lines [0296] To test binding of PRO3790 and reference compounds PRO3599 (anti-CD47xPDL1 AB from Pfizer) and PRO2777 (magrolimab; hu5F9 clone; IgG4) to plasma membranous CD47, cancer cells expressing human CD47 and PDL1 were tested in flow cytometry. [0297] 50,000 cells per well of cancer cell line HT-1080 (from ATCC, cat. CCL-121, double positive for CD47 and PDL1) or HT-29 (from ATCC, cat. HTB-38, positive for CD47) were seeded in a 96-well plate. Threefold serial dilutions of the molecules to test were added at concentrations ranging from 33 nM to 0.6 pM (for PRO2777 on HT-29 and HT-1080 and for PRO3790 and PRO3599 on HT-1080) or from 500 nM to 8.5 pM (PRO3790 and PRO3599 on HT-29 cells) to the plates and incubated for 60 min at 4°C with shaking. Supernatants were removed and detection reagent (goat anti-human IgG, PE conjugated, from Jackson 119915P877PC 23.12.2024 Numab Therapeutics AG ImmunoResearch, cat.109-116-170) was added for flow cytometry analysis. The plates were measured with the flow cytometer Novocyte with a flow rate of 120 µl/min and the geometric mean of the PE fluorescence intensity of single cells was calculated. [0298] The cell-binding potency data are summarized in Table 29. The EC50 as well as maximum binding values for PRO3790 and PRO3599 were normalized to the control antibody PRO2777, which exhibits high affinity binding to human CD47. [0299] Both PRO3790 and PRO3599 showed potent binding to PDL1 co-expressing HT- 1080 cells, and weak binding to HT-29 cells which do not express PDL1. High affinity anti- CD47 molecule PRO2777 showed a PDL1-independent highly potent binding to both cancer cell lines tested. Neutralization of CD47/SIRPα interaction [0300] To test the potency of PRO3790, PRO3599 and PRO2777 to neutralize CD47/SIRPα interaction, binding of SIRPα ligand to cancer cells expressing human CD47 and PDL1 in presence of molecules of interest was tested in flow cytometry. [0301] 50,000 cells per well of cancer cell line NCI-H292 (from ATCC, cat. CRL-1848, double positive for human CD47 and PDL1) or CHO CD47 (in-house cell line, positive for human CD47) were seeded in a 96-well plate. Threefold serial dilutions of the molecules to test were added at concentrations ranging from 100 nM to 2 pM (PRO2777 on NCI-H292 and CHO CD47), from 300 nM to 5 pM (for PRO3790 and PRO3599 on NCI-H292) or from 500 nM to 9 pM (for PRO3790 and PRO3599 on CHO CD47) to the plates and incubated for 60 min at 4°C on a shaker. Supernatants were removed and biotinylated SIRPα (from Sino Biological, cat.11612-H02H1, in-house biotinylated) was added to the plates at a final concentration of 0.5 µg/ml. After 30 min incubation at 4°C, cells were washed and detection reagent (streptavidin, PE conjugated, from BioLegend, cat.405204) was added for flow cytometry analysis. The plates were measured with the flow cytometer Novocyte with a flow rate of 120 µl/min, and the geometric mean of the PE fluorescence intensity of bound SIRPα to single cells was calculated. [0302] The potency data for inhibition of CD47/SIRPα are summarized in Table 30. The IC50 as well as maximum inhibition values for PRO3790 and PRO3599 were normalized to the control antibody PRO2777, which exhibits high affinity binding to human CD47. [0303] PRO3790 and PRO3599 showed potent inhibition of CD47/SIRPα interaction to PDL1 co-expressing NCI-H292 cells, and weak inhibition of SIRPα binding to CHO CD47 cells which do not express PDL1. PRO3790 showed a better potency to inhibit SIRPα binding to NCI-H292 cells than the Pfizer molecule (PRO3599), while the potency was 119915P877PC 23.12.2024 Numab Therapeutics AG comparable for the two molecules when CHO CD47 cells were used. High affinity anti-CD47 molecule PRO2777 demonstrated potent inhibition of SIRPα binding to both cell lines tested. Cell binding to RBCs [0304] To test the binding of PRO3790 and PRO2777 to human red blood cells (RBCs), a flow cytometry-based assay was used. [0305] Human RBCs were isolated from peripheral blood of healthy donors. Human RBCs were seeded in a 96-well plate and 3-fold serial dilutions of the molecules to test were added at concentrations ranging from 100 nM to 1.7 pM (for PRO2777) or from 500 nM to 8.5 pM (PRO3790) to the plates and incubated for 60 min at 4°C with shaking. Supernatants were removed and detection reagent (goat anti-human IgG, PE conjugated, from Jackson ImmunoResearch, cat.109-116-170) was added for flow cytometry analysis. The plates were measured with the flow cytometer Novocyte with a flow rate of 120 µl/min and the geometric mean of the PE fluorescence intensity of single cells was calculated. [0306] The cell-binding potency data are summarized in Table 31. The EC50 as well as maximum binding values for PRO3790 were normalized to the control antibody PRO2777, which exhibits high affinity binding to human CD47. [0307] PRO3790 exhibited a remarkably lower binding potency than the magrolimab analog (PRO2777) to human erythrocytes as indicated by a calculated rel. EC50 of 0.003 fold. Hemagglutination [0308] To test whether PRO3790 triggers agglutination of human erythrocytes, a hemagglutination assay based on optical density measurement was performed. [0309] The hemagglutination assay was performed as follows. Human erythrocytes were isolated from peripheral blood of healthy volunteer and 1 mio. human erythrocytes were seeded in a 96-well plate. Threefold serial dilutions of the molecules to test were added at concentrations ranging from 100 nM to 1.7 pM (for PRO2777) or from 500 nM to 8.5 pM (PRO3790) to the plates and incubated for 120 min at 37°C and afterwards at 4°C overnight. The optical density (OD) of each well was measured at 800 nm using BioTek Synergy Neo2 multi-plate reader. Obtained OD values were plotted as function of log-transformed concentrations and EC50 values were calculated by using GraphPad prism software. g n ) idi n /b a g ) i n b m a / il di n /mi l e b l epo r ) ) b l epl p mumg Mn Mn mmo r g ma ma i sa xI 0 0 ua sa sI F m I 00 mi IF m I F a m. M . F M 2 l . t a 2 ex a x ( t a x ( a F M m M . M. 0 0 . xa Rm ( a 7 m 1 5 . 0 9 5 . x . 0 l ea x Rm ( a . 0 m 0 . 1 0 7 . 8 1 2 7 m (gn ) i l db a l s e i nmi s b l ll ] c mo r ge M 7 4 ] ua c 9 n D M m m I 2- [ T 0 5 7 9 6 0 0 0 0 C n O [0 5 68. 9 i . xaF M H C E 2 . 0 1 > 1 > H C C 6 E 0 . 9 9 0 0 1 4 1 m . l . e x Ra 8 m 1 1 . 0 g ) ) i n b gi b ) d /a n /a l e in l emil di n mil p bp l ep m a mumo r b o r s as g a mumag a 0 5 mi xI aF m I mi sI C F m I E m . M . F x le xM a M . F M / a. b x m . x Rm a. ( a m 1 5 4 . 0 2 4 . 0 l e x a Rm . ( a 0 m 0 1 52 . 9 95 8 mil o sll 0 r 5 g a 1 ec se C 4 E m0 3 1sl l 2 ] t M y c . l eC5 0 0 e 9 2 n [ o r RE ( 1 . 0 c 0 ] H 8 0 M I - 0 5 2 n C C 7 8 0 . 64 0 3 2 h t . y r ] 1- [ T 0 5 1 703 N E 0 . 0 0 2 5 1 9 2 e n a Mn H C E . 0 . 0 . 0 m [0 5 5 6 n io n u C 2 3 tc i o . . t H E 0 78 9 r 3 a r 3 e 0 c 0 an s e t re c s- i t 3 di n b a - 4 b im 7 l 0 n i d b α i n a 4 0 l ug re 0 b c s- 4 o r B-6 P b mil or B-6 r g a di n a m4 0 D g 2- r e I R 7 4 g C a m0 7 iz f P / S D a 2 - eiz m e b il or B-6 74 C m0 7 f P h 7 4D g a 2- D & 9 C C m0 7 1. C 1. 9 B re 0 c f o r 0 c R 1 d s- n ed s- o . t 9 re 0 c i n 3 0 i ot i n 3 0 g d s- b1 B L -0 r a b B e i z D . P a . n 2- 73 iz f l a 1 -0 r i e nd i n 3 b 0B P r t LD . a - u e P . n 2- 73 iz f i n 1 P b l L 0 l D . 2 P a . - I 7 0 9 9 9 N e n 7 3 7 : 7 7 0 9 9 C: 7 0 D 7 7 5 0 D 7 7 9 5 1 D 7 7 9 O 2 O3 O3 O 3 le I O 2 O3 O3 O 3 le I O 2 7 O3 R P R P R P R P b a R R R R T P P P P b a R R OR T P P P 119915P877PC 23.12.2024 Numab Therapeutics AG [0310] The data of hemagglutination assay are summarized in Table 32. The EC50 as well as maximum OD values for PRO3790 were normalized to the control antibody PRO2777, which exhibits high affinity binding to human CD47. PRO3790 showed no sign of hemagglutination at all tested concentrations (highest concentration was 500 nM), while the magrolimab analog (PRO2777) was able to trigger agglutination of human erythrocytes with EC50 value of 0.182 nM. Table 32: Hemagglutination Hemagglutination assay Rel. maximum Rel. EC50 hemagglutination PRO ID PDL1 binder CD47 binder EC50 [nM] (EC50 (OD800nm magrolimab / sample / EC50 sample) OD800nm magrolimab) PRO2777 n.a. magrolimab 0.182 1 1 PRO3790 37-20-B03-sc09.1 70-26-B04-sc03 No hemagglutination observed at 500 nM PD-1 / PDL1 neutralization assay [0311] The potency to inhibit PD-1 / PDL1 interaction was assessed in a cell-based reporter gene assay employing human PD-1-expressing Jurkat NFAT reporter cells. In this assay, CHO cells stably expressing PDL1 and a T cell receptor (TCR) activator were incubated with Jurkat T cells stably expressing PD-1 and as a reporter gene to monitor T cell activation, firefly luciferase under the control of the NFAT response elements (target and effector cells both from BPS Bioscience). Upon binding of the TCR activator on CHO cells to the Jurkat T cells, TCR signaling triggers NFAT-induced expression of firefly luciferase. The interaction between PDL1 and PD-1 negatively regulates TCR signaling and decreases firefly luciferase expression. Consequently, blockade of the PDL1/PD-1 interaction in this system restores luciferase activity. [0312] 35’000 CHO/PDL1/TCR activator cells (BPS Bioscience) in 100 µl of cell culture medium (DMEM/F12, 10 % FCS) were added to the inner wells of a white cell culture plate and incubated for 16-20 h at 37°C and 5 % CO2. On the next day, 95 µl of cell culture medium was removed from each well and 50 µl of 2-fold concentrated 3-fold serial dilutions of the multispecific antibody test compounds comprising a PDL1-BD and of reference antibody avelumab ranging from 50 nM to 8 pM (final concentration in the well) were added. 50 µl of effector Jurkat cells (BPS Bioscience) diluted at 400’000 cells/ml in assay buffer 119915P877PC 23.12.2024 Numab Therapeutics AG (RPMI1640 with 10 % FCS) were added to each well, and plates were incubated for 6 h at 37°C and 5 % CO2. Finally, 50 µL luciferase substrate (BPS Bioscience) prepared according to manufacturer’s protocol, were added per well, and plates were incubated 30 min in the dark, whereafter luminescence was measured using microplate reader BioTek Synergy Neo2. Concentration-response curves were fitted by using a four-parameter logistic fit (GraphPad prism software) and the half-maximal effective concentration (EC50) was calculated. For comparison of multispecific antibody test compounds from different assay plates, relative EC50 values were used. These values were determined by calibrating the EC50 value of the molecules against the reference molecule avelumab included on each assay plate (relative EC50: EC50, avelumab / EC50, test). In addition, the maximum activation of each multispecific antibody test compound was calculated and normalized to the maximum activation seen for avelumab (relative maximum activation: maximum activation, test / maximum activation, avelumab). [0313] PRO3790 showed potent blockade of PD-1 / PDL1 interaction in transgenic Jurkat NFAT reporter gene assay, comparable to benchmark antibody avelumab and reference molecule PRO3599 (Table 33). Table 33: PDL1 / PD-1 transgenic NFAT reporter gene assay Rel. EC50 Rel. maximum (EC50 activation (max. PRO ID PDL1 binder CD47 binder EC50 [nM] avelumab / activation sample EC50 / max. activation sample) aveluamb) - avelumab - 0.5070 1.000 1 PRO3599 WO 2021/124073 WO 2021/124073 0.5254 0.965 0.97 PRO3790 37-20-B03-sc09.1 70-26-B04-sc03 0.2720 1.864 0.95 Phagocytosis assay [0314] Human peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats of healthy donors by density gradient centrifugation (STEMCELL Technologies). CD14+ cells were isolated from PBMCs by positive selection with magnetic beads (Miltenyi Biotech, 130-050-201). HMDMs were generated by culturing CD14+ cells for 6-8 days in complete medium (RPMI 1640, 10 % heat-inactivated bovine serum, Sigma Aldrich) in the presence of 50 ng/ml human macrophage colony stimulating factor (M-CSF) (Peprotech) at 37°C and 5 % CO2. Medium was exchanged and M-CSF replenished on day 3 of the culture. 119915P877PC 23.12.2024 Numab Therapeutics AG [0315] HMDMs were harvested from culturing flasks on day 6-8 using Accutase (Biowest), seeded at 50’000 per well in 96-well flat bottom plates (TPP) and treated with 10 ng/ml IL-10 (Peprotech) overnight. The next day HMDMs were co-incubated with CFSE-labelled target cells at an effector:target ratio of 1:1 for 2.5 h at 37°C in the presence of a serial dilution of test compounds (dose range 200 nM – 0.02 pM). In the case of RBCs, the effector:target ratio was 1:50. At the end of the incubation period, cells were stained with a viability marker (Zombie NIR, Biolegend) and an anti-CD14-APC staining antibody (Biolegend). Samples were fixed and acquired by flow cytometry (Attune NXT, Thermo Fisher Scientific). Phagocytic activity was determined as double positive events (CD14+ HMDMs that have taken up CFSE positive target cells) and phagocytosis is presented as % CFSE+ macrophages. [0316] Figure 7 shows effective phagocytosis of tumors with bi-specific compounds necessitates the inhibition of both PDL1 and CD47. In the case of CD47 x PDL1 bi-specific compounds, the phagocytosis curve exhibits a bi-phasic pattern, in contrast to magrolimab, which demonstrated a straightforward dose-dependent phagocytosis. The initial part of the curve depends on PDL1 inhibition, whereas the second part is driven by CD47 blockade. Given that the PDL1 inhibitor is the same across all three tested molecules, the EC50 for this phase remains comparable. [0317] Phagocytosis of tumor cell by PDL1 x CD47 bi-specific antibodies is more pronounced with high affinity CD47 binder (PRO3791) when compared to those with low affinity CD47 moieties (PRO3790 and PRO3802). This is reflected by EC50 of the second part of the curve, highlighting the variation between compounds featuring high and low CD47 binders. Example 5: Anti-CD47 x PDL1x LILRB2 trispecific antibodies 5.1 Manufacture of anti-CD47 x PDL1x LILRB2 trispecific antibodies [0318] The trispecific antibodies according to the present invention have been produced analogous to the methods described in Example 4.1. Briefly, expression of multispecific constructs was performed in ExpiCHO-S cells using the transient ExpiCHO expression system (Gibco). Expression cultures were cultivated in batch using shaking flasks for 8 days at 37°C. The culture supernatants were separated from cells by centrifugation followed by a 0.22 μm sterile filtration. All target proteins were captured from the clarified culture supernatants by protein A affinity chromatography and were polished by a size-exclusion chromatography (SEC) step, except for PRO3851, which was polished by cation-exchange chromatography (CIEX). 119915P877PC 23.12.2024 Numab Therapeutics AG [0319] Some target proteins with a high post-capture monomeric purity (i. e. PRO2518, PRO3483, PRO3603) were not SEC-polished and, instead, dialyzed against the respective final buffer in a dialysis membrane. For the quality control of the manufactured material, standard analytical methods such as SE-HPLC, SDS-PAGE, and UV280 were used. Table 34 shows a production overview of these molecules. All molecules were finalized to a monomeric content ≥95 % after polishing or dialysis, respectively. 5.2 Biophysical characterization of anti-CD47 x PDL1x LILRB2 trispecific antibodies according to the invention Storage stability study [0320] PRO4672, PRO5052, PRO4687 and PRO5055 were subjected to stability studies such as a four-week stability study, in which the scFvs were formulated in an aqueous buffer (20 mM histidine, 125 mM NaCl, 0.02 % Poloxamer 188, pH 6.0) at 10 mg/ml and stored at 4°C, 25°C and 40°C for 2 weeks, 4 weeks and up to 8 weeks. At the minimum, the fractions of monomers and oligomers in the formulation were evaluated by integration of SE-HPLC peak areas after two weeks, four weeks and at the end of each study. Table 35 summarizes stability results obtained at t2w (2 weeks), t4w (4 weeks) and endpoint measurements obtained at t8w (8 weeks) of the study. [0321] As shown in Table 35, PRO4672, PRO5052, PRO4687 and PRO5055 showed good storage stability at 4°C, 25°C and 40°C. The loss of monomeric content after storage at 25°C for 4 weeks or at 4°C for 8 weeks was less than 1 %. More surprisingly, the loss of monomeric content after storage at 40°C for 4 weeks was still less than 5 %. None of the stable molecules showed considerable loss of protein content at any temperature. Freeze-thaw stability [0322] As in the early development stages the drug substance is usually stored frozen, formulation and fill/finish processing will require some freezing and thawing operations. The compatibility of the top performing trispecific molecules with respect to freeze-thawing (F/T) cycles (colloidal stability) was assessed. [0323] For the F/T stability assessment the same analytical methods (SE-HPLC, cGE) and parameters (% monomer content and % monomer loss) as for the storage stability study were applied to monitor the quality of the molecules over five F/T cycles. Table 35 illustrates that after five F/T cycles (F/T) the loss of monomer content was less than 1 %. None of the stable molecules showed considerable loss of protein content at any temperature (data not shown). Ci cr ] Ee -m% s S [ t sony t l i e ooru . 7 7 . . 9 27 9 6 7 . . . . . . . . . 9 88 9 19 9 38 9 29 9 78 29 48 98 7 9 . 47 . 05 . 3 9 . 7 8 . 5 0. 7 0 0 . 2 7 . 6 8 u P m p 9 9 9 9 9 9 9 9 9 9 1 9 9 c l e r o ei t m t e c -e n e t r * ] s u t r e op/ L Pag f c m [ 09 247 234 197 187 236 184 180 1 02 281 107 1 2 364 343 4 3 768 164 2 83 1 85 284 2 e r i c n n i i i i i i f a i a na na na na c e re 70 c m od 70 c m od 70 c m od 70 c m od 70 c m od 70 m od 7070202020 20 7070 p s d i r i n s - y s - y s - y s - y s - y c s - y c s - c s - c s - c s - c s - c s - c s - c s- t b2 1 1 m E mu 1 1 mm1 1 mm1 1 mm 1 1 mm1 1 mm1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 d n B a R -8 d E-8 u d E-8 u d E-8 u d E-8 u d E-8 u d E-8 E- E- E- E- E- E- E-s I L 1- AS 1- AS 1- AS 1- A 1- A1- A1 - 8 1- 8 1- 8 1- 8 1- 8 1- 8 1- 8 1-i 8 e L 6 H8 6 H8 6 H8 6 S H 8 6 S H8 6 S H8 6 8 6 8 6 8 6 8 6 8 6 8 6 8 6 dob n n .t i t i a i a I n n a 33333333 33 mm ( i c r 0 0 0 0 0 0 0 0 0 0303 od od 32 0 3030 i f e c d s - c s - c s - c s - c s- c c e i n 4 0 4 0 4 0 4 0 4 s - c 0 4 s - c 0 4 s - c 0 4 s - c 0 4 s - c 0 4 s- 0 c 0 4 s - y y 0 c 0 4 0 mm s- c s c 40-5 s - c s- 0 4 0 4 0 p s b B i r 7 4 -6 B- t D 2 - 6 B- 0 7 2 - 6 B- 0 2 - 6 B- 2 - 6 B- 2 - 6 B- B - - B - - B - - B - - B m - - u mu B - - C- ) B- B- 7 0 7 0 7 0 7 2 - 6 07 2 6 0 2 6 0 2 6 0 2 6 0 2 6 0 2 dA dAr e r e 6 0 S S zif iz f 2 - 6 03- 7 0 46 7D2 - 6 07 2- 07 6 A C 7 7 7 7 7 7 H H P P 7 C 4S 1h/ 2 1.
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[ 3 2 5 5 6 5 4 5 5 4 7 5 G [) cD sr BC ( 95 1 50 7 06 2 53 8 6 . 2 8 . 4 06 3 69 8 95 1 52 48 . 1 0 . 4 95 1 50 7 on w t 8 . 1 0 - . 60 - . 50 - . 2 o 0 - d 2 ( + y R . 4 . 0 . 3 1 . 2 . . . 1 . 8 . . 5 . . - 2 ± 4 ± 1 ± ± 3 3 2 9 7 ± 3 ± 2 ± ± 2 3 1 9 4 . 40 ± 4. 1 1 2 4C e ° k D-A4 1 . 9 4 w t 44. 0 . 20 - . 20 - . 30 - no L HD C7 . 3 40 . 9 2 . 1 1 5 2 . 1 59 . 1 93 . 2 9 . 7 4 5 0 . 1 8 ± 6 . 3 8 ± ± 5 6 9 . . 8 8 ± 9 . 1 9 ± 7 . 3 40 9 Ms C u ° . . 5 2 w t 44. 0 . 4 0 - . 6 0 - . 4 l g + 2 . 0 - . o 8 32 1 2 . 7 07 1 7 . . 4 35 1 3 . 46 . 8 0 . . 4 72 2 . 3 . 2 . . 8 32 1 s mR C o D-+± A4 1 6 ±1 ±1 ±5 5 ±1 ±1 ±8 ±9 ±2 5 08 ±6 ±1 M n y LD. 2 . 2 4 . 4 1 . 3 8 ± 3 . 4. 7 1 . . . . ± ±. . 0 8 2 1 40 2 1 . 7 . 9 2 4 w t 4 . 11 - . 42 - . 44 C H C 1 2 1 2 8 1 1 2 1 1 2 1 4 6 7 2 1 4 2 - . 22 - B Po C ° n 0 y ) +3 4 w t 2 . 2 0 - . 0 1 - . 2 2 - . 3 1 - cd D n C ( 2 . . 3 1 . . 3 2 . 3 1 . 8 1 . 6 2 2 2 . 3 a l s 0 2 / T ± 0 7 ± 0 . 0 . 0 6 . 0 0 . 0 . 0 . 0 2 F 5 1. 0 . 30 - . 40 - . 30 - n a l e ±. ±. ±±± ± . ± ± . ±±± ± . 7 m c 6 u T . 1 72 3 1 . 5 75 5 7 . 0 8 . 0 3 . 7 46 5 1 . 1 62 1 5 . 0 7 . 0 6 . 1 72 3 4 1 . . . . h w t 4 3 0 0 8 8 9 8 9 8 9 9 9 o t s i e ) 5 . 0 . 1 . 1 . 4 . 4 . 0 d ] l s C l +0 2 . 0± 4 8 ±. 0± 4 1 ±. 0 1 ± . 0± 1 ± 5 5 ±. 0± 2 1 ±. 0 1 . 0 5 . 0 4 ± ° 4 w. t 9 4 8 . 9 78 . 9 38 . 9 9 o 89 bi t 0 0 e 0 cD9 n . . 03 4 . . 1 8 0 . 8 . 0 8 . 3 ±6 ±0 ±9 . 0 a 1 BC ( 3 3 7 0 4 . 0 . 1 0 1 5 4 . 3 6 1 . 0 . 1 . 3 3 3 C ° . 9 . . . 2 x IF 5 2 w t 4 8 9 5 8 9 9 7 9 8 8 9 BR M IL o e +R . 37 . 57 . 2 . 2 45 . 8 . 0 2 . 5 7 7 . 3 . 5 L G [ D - - ± ± 3 4. 6 . 6 6 2 . 3 4. 4 . 5 7 7 x ) A4 1 . . ± . ± . ±± ± . ± . ± ± . ±± ± . ± w. t 4 4 7 . 9 56 . 9 04 . 9 07 9 1 s L ro L HD4 C3 1 61 4 53 1 01 42 . 8 2 . 8 20 2 96 41 . 9 05 2 9 . 5 3 . 7 43 . 1 61 4 D nod C ° P 2 + . 0 . . 1. 6 . 4 1 41. 4. . 8 2 6. 9. 1. 4 w t 3 2 8 . 9 97 . 9 36 . 9 97 9 x 7 4 ( R 4 5 ± 0 5 ± 1 1 1 1 . 1 7 2± . 1 1 8± . 8 7 0 1 . 4 5 6± D n a / C T - mD-+ u A4 L1 ± D. 0 1 . 5 5 ± 4 . 0 0 . 0 9 ± 7 . 6 ± 6 . 7 ± 2 . 4 9 . 6 8 ± 8 . 9 3 . 6 8 ± 0 . 2 ± 6 . 4 ± . 0 1 . 5 . 6 . 6 . . H H C6 1 6 1 3 5 7 1 41 2 0 4 5 6 4 1 F 5 8 9 8 9 1 8 9 9 8 9 i t naf ] 0 . o -Mn [ t 5 8 . 9 9 8 . 9 5 8 . 9 2 9 g n 9 i nd e cni i n noot a r t 6 . 100 2 6 . 100 2 6 . 100 2 6 . 100 2 6 . 100 2 6 . 100 6 . 00DI 7 5 8 5 B C 2 1 2 2 2 7 5 : - 6 O 4 0 O5 6 O4 0 5 7 3 2 le D I 5 3 0 5 4 5 0 5 5 6 7 4 5 0 5 5 0 5 5 0 5 5 0 3 5 1 5R R R OO O O O O O O O O P P P R P R P b a R R R R R R R R T P P P P P P P P 119915P877PC 23.12.2024 Numab Therapeutics AG 5.3 Pharmacodynamic characterization of anti-CD47 x PDL1 x LILRB2 trispecific antibodies according to the invention Binding affinity to human CD47, PDL1 and LILRB2 [0324] Affinity of trispecific molecules to human and cynomolgus CD47, and human and cynomolgus PDL1 was determined by SPR analysis as described in Examples 2.6 and 3.6. For affinity measurements to human LILRB2, a different SPR measurement procedure was applied to determine the binding affinity without avidity effects. [0325] Briefly, binding affinity without avidity was determined by using a proprietary anti- framework rabbit IgG (rIgG21-57-20-B11) immobilized to a carboxymethylated dextran surface (CM5 sensorchip; Biacore, Cytiva) to capture the multispecific antibodies according to the present invention on the surface. For each antigen, a titration series of the respective antigen was injected as analyte. After each capture-analyte injection cycle, every flow channel on the sensor chip was regenerated (3 M MgCl2), and a multispecific antibody according to the present invention was captured again followed by a different concentration of the antigen. The binding kinetics to the human antigens were measured using a multi- cycle kinetic assay, with eleven analyte concentrations ranging from 0.088 to 90 nM, 1:2 diluted in running buffer (HEPES, 150 mM NaCl, 0.05 % Tween-20, pH 7.5; Bioconcept). [0326] The results are shown in Table 36. Binding of trispecific antibodies to different human and cyno PBMCs cell types [0327] Binding of the multispecific antibodies of the invention to different PBMC cell types from human as well as from cynomolgus monkeys was examined within the specified subset of T cells, B cells, and myeloid cells. In particular, the ability of the anti-CD47 x PDL1 x LILRB2 antibodies to target HLA-DR+ CD14+ monocytes and HLA-DR+ CD14- APCs, both from human and cynomolgus monkey, was investigated. The method used to assess the binding to these cell-types was analogous to the method described in the preceding section. The results are summarized in Table 37. Binding is detected by an increase in fluorescence signal indicated in [GeoMFI x 1000]. Re-polarization of M2 macrophages and LPS-driven cytokine secretion [0328] The ability of the anti-CD47 x PDL1 x LILRB2 antibodies to re-polarize M2 macrophages and to foster LPS-driven cytokine secretion was assessed using the methods disclosed in Example 3.6. 119915P877PC 23.12.2024 Numab Therapeutics AG Table 36: Affinity of different multispecific test constructs to human and cynomolgus CD47, human and cynomolgus PDL1, and human and cynomolgus LILRB2. Protein ID Target Affinity ka (M-1 s-1) kd (s-1) KD (M) PRO4672 human CD47 4.48E+05 2.56E-02 5.72E-08 human PDL1 1.05E+06 1.26E-04 1.20E-10 human LILRB2 *) 2.4E+05 8.9E-05 3.7E-10 PRO4687 human CD47 8.11E+05 2.04E-02 2.52E-08 human PDL1 1.31E+06 1.16E-04 8.85E-11 human LILRB2 *) 2.6E+05 9.3E-05 3.6E-10 PRO5052 human CD47 6.6E+05 2.2E-02 3.4 ±0.2E-08 cynom. CD47 3.2 ±0.1E-08 human PDL1 2.3E+06 2.9E-05 1.3 ±0.4E-11 cynom. PDL1 1.2 ±0.2E-11 human LILRB2 *) 2.9E+05 5.4E-05 1.9 ±0.8E-10 cynom. LILRB2 *) no binding PRO5053 human CD47 5.33E+05 2.65E-02 4.97E-08 human PDL1 2.05E+06 2.72E-05 1.33E-11 human
Figure imgf000150_0001
PRO5055 human CD47 5.1E+05 1.1E-02 2.1 ±0.04E-08 cynom. CD47 2.1 ±0.1E-08 human PDL1 2.2E+06 3.3E-05 1.6 ±0.7E-11 cynom. PDL1 8.4 ±0.9E-12 human LILRB2 *) 2.6E+05 9.2E-05 3.5E-10 cynom. LILRB2 *) no binding PRO5056 human CD47 4.39E+05 2.49E-02 5.66E-08 human PDL1 2.02E+06 2.13E-05 1.05E-11 human
Figure imgf000150_0002
*) Affinity to human and [0329] It was observed that the anti-CD47 x PDL1 x LILRB2 antibodies of the invention could inhibit LILRB2, causing a dosage-dependent rise in the M1 marker CD86 and a decline in the M2 markers CD163 and CD206, with a similar potency to that of the monospecific anti-LILRB2 IgG4 PRO4710 (see Table 38). On the other hand, the reference anti-PDL1 x CD47 bi-specific antibodies from Pfizer (PRO3599), the LILRB2-BD-devoid trispecific variants PRO5053 and PRO5056 (HSA-BD as LILRB2 mock binding domain), as well as the anti-CD47 antibody magrolimab and the anti-PDL1 antibody atezolizumab (tested 119915P877PC 23.12.2024 Numab Therapeutics AG at a 50 nM dose), showed no re-polarization activity. Results of the cytokine secretion are summarized in Table 39. Table 38: Re-polarization potency of M2 macrophages for the anti-LILRB2 IgG PRO4710, the trispecific antibodies PRO4672, PRO4687, PRO5052, PRO5053, PRO5055 and PRO5056, and reference compounds magrolimab, atezolizumab, and PRO3599 PRO ID CD86 EC50 [nM] CD163 EC50 [nM] CD206 EC50 [nM] PRO4710 0.119 ± 0.070 0.072 ± 0.008 0.203 ± 0.129 magrolimab n.a. n.a. n.a. atezolizumab n.a. n.a. n.a. PRO3599 n.a. n.a. n.a. PRO4672 0.079 ± 0.009 0.067 ± 0.004 0.120 ± 0.043 PRO4687 0.078 ± 0.009 0.070 ± 0.009 0.124 ± 0.077 PRO5052 0.082 ± 0.006 0.073 ± 0.001 0.134 ± 0.082 PRO5053 n.a. n.a. n.a. PRO5055 0.085 ± 0.012 0.058 ± 0.003 0.128 ± 0.071 PRO5056 n.a. n.a. n.a. [0330] The trispecific antibodies according to the invention further exhibit the ability to foster LPS-driven cytokine secretion (see Figures 8 and 9, and Table 39), due to the presence of the LILRB2 arm, which is confirmed through comparison with the respective mock compounds. The tested multispecific antibodies demonstrate similar effects obtained with the reference anti-LILRB2 antibody J19.H1 from Jounce (PRO2518, see Figure 9), while there is no observed activity for the LILRB2-BD-devoid trispecific variants PRO5053 and PRO5056 (HSA-BD as LILRB2 mock binding domain) and the reference antibodies PRO3599, magrolimab and atezolizumab (at the top dose). [0331] Both assays demonstrate the efficacy of LILRB2 inhibition through the absence of activity in PDL1 x CD47 bi-specific compounds, and Pfizer compound. Furthermore, comparable activity is observed between multispecific constructs comprising a high-affinity CD47-BD and a low-affinity CD47-BD. Notably, the trispecific antibodies of the invention having different PDL1-BDs exhibit an activity comparable with established benchmarks from Jounce, Merck, and NGM. 119915P877PC 23.12.2024 Numab Therapeutics AG Table 39: Induction of cytokine secretion for the anti-LILRB2 IgG PRO4710, the trispecific antibodies PRO4672, PRO4687, PRO5052, PRO5053, PRO5055 and PRO5056, and reference compounds magrolimab, atezolizumab, and PRO3599 PRO ID IL-1β EC50 [nM] TNF-α EC50 [nM] IL-6 EC50 [nM] PRO4710 0.248 ± 0.136 0.169 ± 0.026 0.153 ± 0.043 magrolimab n.a. n.a. n.a. atezolizumab n.a. n.a. n.a. PRO3599 n.a. n.a. n.a. PRO4672 0.096 ± 0.026 0.123 ± 0.037 0.078 ± 0.007 PRO4687 0.205 ± 0.166 0.175 ± 0.038 0.097 ± 0.020 PRO5052 0.082 ± 0.028 0.117 ± 0.052 0.077 ± 0.024 PRO5053 n.a. n.a. n.a. PRO5055 0.0705 0.141 ± 0.066 0.063 ± 0.019 PRO5056 n.a. n.a. n.a. 5.4 Phagocytosis [0332] The in vitro phagocytic activity induced by bi-/multispecific antibodies of the invention was assessed using the H292, A375, Nalm6, tumor cell lines and primary red blood cells (RBCs). The assay relies on the uptake of labelled target cells by macrophages and analysis by flow cytometry. [0333] Human peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats of healthy donors by density gradient centrifugation (STEMCELL Technologies). CD14+ cells were isolated from PBMCs by positive selection with magnetic beads (Miltenyi Biotech, 130- 050-201). HMDMs were generated by culturing CD14+ cells for 6-8 days in complete medium (RPMI 1640, 10 % heat-inactivated bovine serum, Sigma Aldrich) in the presence of 50 ng/ml human macrophage colony stimulating factor (M-CSF) (Peprotech) at 37°C and 5 % CO2. Medium was exchanged and M-CSF replenished on day 3 of the culture. [0334] HMDMs were harvested from culturing flasks on day 6-8 using Accutase (Biowest), seeded at 50’000 cells per well in 96-well flat bottom plates (TPP) and treated with 10 ng/ml IL-10 (Peprotech) overnight. The next day HMDMs were co-incubated with CFSE-labelled target cells at an effector:target ratio of 1:1 for 2.5 h at 37°C in the presence of a serial dilution of test compounds (dose range 200 nM – 0.02 pM). In the case of RBCs, the effector:target ratio was 1:50. At the end of the incubation period, cells were stained with a viability marker (Zombie NIR, Biolegend) and an anti-CD14-APC staining antibody (Biolegend). Samples were fixed and acquired by flow cytometry (Attune NXT, Thermo 119915P877PC 23.12.2024 Numab Therapeutics AG Fisher Scientific). Phagocytic activity was determined as double positive events (CD14+ HMDMs that have taken up CFSE positive target cells) and phagocytosis is presented as % CFSE+ macrophages. [0335] The results of the phagocytosis potency analysis are summarized in Tables 40, 41, 42 and 43 as well as in Figures 10, 11, 12 and 13.The multispecific antibodies induced a bi- phasic response curve, with an initial part driven by the anti-PDL1 arm, confirmed through comparison with avelumab (anti-PDL1 IgG1). This initial increase in phagocytosis was absent in PDL1-negative cell lines, confirming PDL1 dependency. The second part of the dose-response curve is driven by anti-CD47 engagement, which was confirmed by PRO5054 and PRO5057, two compounds in which anti-CD47 was replaced by an unspecific human serum albumin (hSA) binder (hSA-BD), herein also referred to as mock binding domain. In PDL1-positive cell lines the herein described antibodies showed similar potency but higher efficacy as the benchmark compounds magrolimab, PRO3599 (anti-CD47xPDL1 bispecific antibody of Pfizer), PRO3851 (anti-CD47xPDL1 bispecific antibody of Innovent) and a combination of magrolimab & avelumab. In PDL1-negative cell lines potency was reduced compared to benchmark compounds, highlighting the PDL1 dependency of the described antibodies. Modification of the Fc-effector functions (PRO5134 and PRO5423), herein also referred to as Fc-silent version, abolished phagocytosis. [0336] As apparent from Figure 11 and the data in Tables 40 and 41, the trispecific antibodies of the invention PRO5052, PRO5055, and PRO4672 surpasses the efficacy of the magrolimab and avelumab combination in phagocytosis. PRO4687 showed high efficacy against H292 cell but not against Nalm6 cells. For the comparative molecules PRO5054 and PRO5057, where the CD47-BD has been replaced by a mock binding domain, significantly reduced phagocytosis potency for H292 and A375 cancer cells could be observed, as expected. [0337] From Table 42 and Figure 12, it is further evident that phagocytosis potency may also be increased by the presence of the LILRB2-BDs, as indicated by the somewhat reduced activity observed in the trispecific molecules, where the LILRB2-BDs have been replaced by mock binding domains (PRO5053 and PRO5056). [0338] From Tables 42 and 43, it is further evident that re-formatting the multispecific antibodies by placing the anti-LILRB2 domains in the Fab arms, opposite of the Fc-region, completely abolished tumor cell phagocytosis (PRO4677). Combination of magrolimab (PRO2777) with PRO2734, an anti-LILRB2 IgG4 antibody (Merck), did not enhance phagocytosis, indicating that the activity/efficiency of the LILRB2-BDs is also significantly format-dependent, and not only signalling-dependent. It is believed by the inventors that LILRB2-engaging domains in proximity to the Fc-region improve orientation of the 119915P877PC 23.12.2024 Numab Therapeutics AG compounds enhancing trans-bridging of macrophages and tumor cells (in absence and presence of PDL1 expression on tumor cells). [0339] Many different multispecific antibody formats have been tested for H292 cancer cell phagocytosis and red blood cells (RBC) phagocytosis (see Table 43 and also Figures 10, 11, 12, 13, 14 and 15). The data illustrate that the following multispecific antibodies and reference antibodies exhibit high levels of cancer cell phagocytosis, but at the same time also significant levels of RBS phagocytosis: - trispecific antibodies PRO3843, PRO4682, PRO4683 and PRO4711, where the CD47- BD is located in one of the Fab arms in the form of a Fab; - bispecific antibodies PRO3609 and trispecific antibody PRO3613, where the CD47-BD has an intermediate binding affinity for CD47; - the reference anti-CD47 x PDL1 antibodies PRO3599 (Pfizer), PRO3851 (Innovent) and magrolimab. On the other hand, the bispecific antibody PRO3790 and the trispecific antibodies PRO4672, PRO4673, PRO4687, PRO4688, RO5052, PRO5053, PRO5055 and PRO5056 exhibit high cancer cell phagocytosis and, at the same time, only minimal or no measurable RBC phagocytosis. The bispecific and trispecific antibodies of the invention thus have a significantly larger therapeutic window than the reference antibodies PRO3599, PRO3851 and magrolimab. The phagocytosis data in Table 43 further illustrates that placing the CD47- BD at the C-terminus of the Fc-region (PRO4677, PRO4678) or replacing the CD47-BD by a mock domain (PRO5054), also lead to minimal or no measurable RBC phagocytosis, but also dramatically decrease cancer cell phagocytosis. Generally, all tested multispecific antibodies of the invention containing a CD47-BD in scFv format in one of the Fab arms did not induce noteworthy RBC phagocytosis at all concentrations tested and, thus, exhibit a large potential therapeutic window, as illustrated in Figure 15. Phagocytosis of PDL1 positive tumor cells was unaffected by the different format and positioning of the CD47-BD, except for PRO4677, where tumor and RBC phagocytosis were abolished. 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- v a - v a 3 3 c s 3 3 c s 3 3 c s 3 3 c s s o s o t y e c e v t y n a a c o c og d ) g a a h C n : 2 a b a h + p p b + + + m u 4 4 3 7 l e 7 2 D I 7 mi 7l 4 3 2 o r 7 2 ) k 7 7a 7 7 4 3 7 7 4 3 b a 2 5 3 5 5 5 6 5 m u c7 2m7 2 7 2 7 2 7 2m0 5 0 5 0 5 0 5 i m i mx b a O OR ORg aORr eOu Rl eOROROOul e O O O Ox aa M TR P P Pm ( PM ( Pv a P PR PR Pv a R P R P R P R P M) *) * * . s i s i e s e) oi v * ) d m t a 7 * 7 * )* )* 8. 1 3. 1 1 * * . 6. 7. 1. 7. 5. 7.o u t y i t i l mc oe 7 r 2 l O 0 0 5 ± 0 60 10 1 0 0 0 00 00b g R ) ± ± ± ± ± ± ± ±n x a a e v P * * 5 0 8 6 a Mh p l eo t 1 . 1 . 0 9 8 . 0 8 8 . 0 6 9 . 0 4 9 . 0 2 9 . 0 2 9 . 0 5 2 . 0 2 1 . 0 9 1 . 0 1 1 . 0 1 1 . 0 2 0 . 0 e c n e n r e e v 30 42 4 9 7 8 9 . f e i r ] M . 0 6 4 6 . 0 . 3 . 1 . 3 8 1 2 r s dn d C 7 [ 0 . 5 2 . 9 1 . 8 2 0 3 7 5 ±4 ± n B 4DC0 5 ± 1 0 ±0 ±0 ± 1 ±3 0 ±8 7 8 6 ± 5 2 . 4 . 3 a . 4 . a . a . a . a a R C E . 0 . 3 . 3 . 4 . 0 . 2 . 2 . 6 0 1 . n 9 3 . n . n . n . nnoi tne s v i s 8 9 7 6 i n e mo t . 7 . 0 2 1 uy c 6 6 . 7 4. 4 1 8 .6 . 2 54. 3 5 1 5 .5 2. 9 5 1. 5 7 . 5. 96. . i mo)*l ±7 ±6 ±8 1 ± 6 6 ±4 ±5 ±7 ±6 )* ± 3 1 1 8± 1 9 ± 3 5 1 h tfo x a g ahe v . 3 ) . 8 ) . 5 ± . 3 . 1 . 9 s M p l e 9 2 * * 7 3 * * 2 3 9 3 8 4 4 . 3 4 7 . 3 9 1 . 3 1 * 2 . 3 5 3 7 ± 2 . 4 1 5 . 3 0 6 . 3 6 ± 8 2 8 6 . 8 i edo n bi t e v 2 5n i r ] 5. 1 4. 4 6. 4 1. 6 5. 6 5. 7 5. 1 . 8 3 0 . 6 0. 9 . 9 3 3 2 . 9 . 1 a dM i c 7n 4 [ 0 5 1 1 0 . 9 0 5 ± 8 0 2 ± i f DC 2 2 ± c e C E . 0 3 6 ± ± ± ± ± ± 1 1 2 ± . . ± . ± 6 1 3 . ± ± . 8 2 8 . 1 4 2 . 4 3 1 . 2 3 . 9 3 3 . 5 6 7 . 6 2 0 1 2 1 3 2 3 . 8 9 0 1 2 6 . 8 5 3 1 . a . npi s l tu s ne 1 4 6 61 2 7 2 5 l l v] 0 4 2 2 9 0. 2m . 3 . 0 . 2 . 1 . 0 . 0 . 0 . 0 . 2 . 1 0 . f e c i rdMn 0 0 0 0 0 0 0 0 0 0 ± 0 0 6o 2 9 1L [0 5 ± 5 3 . a ± 2 1 ± 5 2 ± 4 1 ± 9 1 ± 1 1 ± 8 0 ± 4 0 ± 2 3 ± 8 3 4 ± 8 . 5 1 s i 2 s H D PC E - . 0 . n . 0 . 0 . 0 . 0 . 0 . 0 . 0 . 0 1 . 0 0 . 0 0 . 0 a . n o t y c 20 70 20 20 7 7 7o g c a D s - c c c 0 c 0 c 0 ch B 1 s- 1 s- 1 s- 1 s- 1 s- 1 s- 1 p - 2 1E 1 1 1 1 1 1 - E- E- E- E E EC BB R 8 1- 8 1- 8 1- 8 - 1 8 - 1 8 - 1 8 1 R I L L 8 8 8 - 8 - - - - - - - 6 6 - 6 6 - 8 6 - 8 6 - 8 6d n a 2 0 2 0 3 3 3 3 3 3 3 3s i s b a c s-5 c s 0 - c 5 s- 0 c 4 s- 0 c s- 0 c s- 0 c s- 0 c s- 0 c s- 0 c s-o t D y c B- im o 7 l 9 9 1 0 0 g 4 o r 5 5 3 8 C C 0B 4 0B 9 9 9 4 5 9 0B 4 0B 4 0B 4 0B 4 0B 4 0B O 3 O -6 -a D g a R R 3 - 6 - 0 3 - 6 - 0 2 - 6 0 2 3 5 - O 3 O -6 - 0 R R 2 - 6 - 0 2 - 6 - - - 0 2 - 6 2- 6 2- 6 2-h C m P P 7 7 7 7 P P 7 7 0 7 0 7 0 7 0 7pll 1 e c . 9 1. r 0 9 1. cs 0 c 81 c 81 c 81 9 c 0 c 81 c 81 c 81 c 81 c 81 c e c -n D 3 0 s- 30 s- 30 s- 30 s- 30 s- 30 s- 30 s- 30 s- 30 s- 30 s- 30 a c B 9 - 9 1 1 5 5 3 8 3 B-0 B-0 B-0 B-0 B 9 - 9 0 5 3 B-0 B-0 B- B- B- B-2 L 9 2 D O P - R O P R 2- P 7 3 2- 73 2- 73 2- 73 2- O 73 R 2- P 7 3 2 - 0 73 2 - 0 73 2 - 0 73 2 - 0 73 2- 73 H: 3 7 4 7 9 9 1 5 9 0 3 1 2 8 3 8 1 1 3 4 0 9 2 3 7 8 7 l e D I 7 5 8 6 6 6 6 7 8 7 7 6 7 6 8 6 8 6 7 6 O 2 O 3 O 3 O 3 O 3 4 4 4 3 3 4 4 4 4 4 b a R P R P R P R P R O P R OR OR OR OR OR OR OR OR OR OR T P P P P P P P P P P P )* * )* * 2 )* * )* )* 3 1 0 4 *5 * . 00 . 00 . 0 . 0 . 0 1. 2 ± 0 ± 0 ± 0 0 4 1 ± . 0 4 1 . 0 3 0 . 0) * * 1 1 ± . 0 2 1 . 0 . d . n . d . n . d ± . n 4 8 . 0 7. 1 2 3 . 8 7. 6 ± 6 9 9 ± ± . 7 9 3 . 4 a . n . a . . n 6 4 4 . a . n . d . n . d . n . 3 d . . n 4 7 3 8 . 3 1 4 6 . 4 9 . 9 . 0 . 1 04 ± 5± 7 5 /- . 1 7 4 5 ±7 ±2 + ± ) * . * 9 4 4) * . * 0 0 2) * . * 7 6 4) * . * 3 3 43) * . * 1 1 2) * * . 7 a . n . a . . n 0 4 3) * * 6 . 5 5 7 9 1 . 2 6 8 1 . 5 0 . 5 3 . 5 ± 3 1 1 1 ±5 ±4 ±0 ± . 9 0 . 1 2 8 2 . a . . n 9 0 . 1 3 4 5 . a . n . a . n . 4 a . . n 2 5 2 2 . 3 . 2 8. 0 . 3 4 4 8 . 1 . 0 0 0. 0 . 0 0 0 0 0 /- ± d 1 e 2 t 7± ± . 0 9 1 ± . 0 9 0 ± . 0 9 1 ± . 0 3 1 + . 0 3 1 a .0 . a . n . a . n 7 0 . 0 t s 5 1 ei s 7 7 7 7 7 7 . w r 0 c s- 0 c 0 c 0 0 0 70 dee h 1 1 s-E 1 1 s - c 11 s - c 11 s - c 11 s - c 11 s- t 1 c t 1 e o ps -8 E - -8 E-8 E-8 E-8 E-8 E- x e s e l n 1 8 6 - 1- 86 1- 86 - 1- 86 1- 86 1 - 8 86 1 - s 8 C u 6 BM Rf n 3 3 3 3 3 3 o0 . 4tM 0 c s- 0 c 4 s- 0 c s- 0 c s- 0 c 0 c 30 c i ss o a n d0 8t 0 4 0 4 0 4 0 s- 40 s- 40 s- 4 t yei n a B-6 B-2 - 6 B-6 B-6 B- B 0 - B c - o gmr e d ei n 0 7 2- 07 - 2- 07 2 - 6 07 - 2 - 6 07 2 - 6 07 2 - a 07 h p t e o dmr n e i s s a t e wl d l42 e e v c 42 c 42 c 42 c 42 c 42 c 42 42 81 r e e v le s-2 0 s- 2 s- 2 s- 2 s- 2 s - c 2 s - c 2 s - c 2 s- h t le 3 s ai s i sG 0 0 0 0 0 0 0 0 - , s o s 3 G- t o t y 0 - 3 G-3 3 0 - 3 G- 3 3 0 - 3 G- 3 3 0 - 3 G- 3 3 0 - 3 G- 3 3 0 - 3 G- B- d 3 3 0 ey - 3 c 3 3 0 - 0 33 2- 7 i nc oog 3 mr e g a a h th2 e p p 5 3 d m u 0 5 4 5 0 5 5 6 7 4 3 4 m 0 5 0 5 0 5 3 2 1 t oui mO 5 O 5 O 5 O 5 0 O 5 1 O 5 4 O 5 9 i O 4 O Nmx a x a R P R P R P R P R P R P R P R P R : . P . d M N M) *) * * 119915P877PC 23.12.2024 Numab Therapeutics AG 5.5 Multispecific antibodies activate NK cells to tumor cell killing via ADCC [0340] The in vitro cytotoxic activity of primary NK cells induced by the multispecific antibodies according to the invention was assessed using the H226 tumor cell line, transfected with the IncuCyte NucLight Orange Lentivirus Reagent (Sartorious, 4625). The assay makes use of the IncuCyte live imaging platform (Sartorius) and quantifies the killing of H226 NucRed tumor cells as measured by the loss of the NucRed signal over time. [0341] Human peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats of healthy donors by density gradient centrifugation (STEMCELL Technologies). NK cells were subsequently isolated by negative selection with magnetic beads (Miltenyi Biotech, 130-092-657). NK cells were pre-activated overnight in (RPMI 1640, 10 % heat-inactivated bovine serum, Sigma Aldrich) in the presence of 0.2 – 1 nM human IL-2 (R&D Systems) at 37°C and 5 % CO2. [0342] H226 NucRed tumor cells were seeded at 7’000 cells per well in 96-well flat bottom plates (TPP) on day -1 and cultured in RPMI+10 %FCS. The next day, pre-activated NK cells were harvested, washed with medium and co-incubated with target cells at an effector:target ratio of 5:1 for 24 h at 37°C in the presence of a serial dilution of test compounds (dose range 20 nM – 0.03 pM). Images of the tumor cell - NK cell co-culture were taken every two hours and the count of H226 NucRed tumor cells was assessed over time. Data is presented as fold change to T0 for each compound and is an average of four images per well. [0343] The trispecific antibodies PRO5052 and PRO5055 exhibited a potent ADCC with EC50 values that are similar to that of avelumab and higher than that of the reference PRO3599. Magrolimab (anti-CD47 IgG4) and atezolizumab (anti-PDL1 modified IgG1) failed to induce ADCC. Also, the Fc-silent variant PRO5134 failed to induce ADCC. The trispecific antibody PRO5054, where the CD47-BD is replaced by a mock binding domain, shows a significantly reduced ADCC, highlighting the importance of simultaneous targeting of PDL1 and CD47. The presence or absence of LILRB2-BDs in the trispecific molecules had no effect on ADCC potency. 5.6 Induction of T cell proliferation [0344] To evaluate whether the trispecific antibodies of the invention are capable to activate T cell proliferation, these antibodies were tested in an immunosuppressive assay. The immunosuppressive assay used aims to track recovery of T cell proliferation in the co-culture 119915P877PC 23.12.2024 Numab Therapeutics AG with suppressive macrophages in the presence of T cell stimulation and test compounds. The assay was performed as follows. [0345] T cells were isolated from PBMC fraction with a magnetic Pan T Cell negative isolation kit (Miltenyi Biotec, cat no.130-096-535). Bulk T cell population was used containing CD4+ and CD8+ T cells. T cells were stained with a cell proliferation tracking dye (CTV CellTrace dye, Thermo Fisher Scientific, cat no. C34557) by incubation of T cells with the CTV dye (5 µM working solution) for 20 min at 37°C. To stop the labelling reaction, 5x volume of complete medium containing 10 % FBS was added for 5 min at room temperature followed by extensive washing. [0346] T cells were stimulated to proliferate by a co-culture in 96 flat-bottom well plates (TPP) with coated anti-CD3 (at 2 ug/mL, BioLegend, cat no.317326) and soluble anti-CD28 mAbs (4 µg/mL, BioLegend, cat no.302934). Suppressive human monocyte-derived macrophages (HMDM) were generated by treatment with IL-10 (10 ng/mL, BioLegend cat no. 571004,) and TGF-β (20 ng/mL, Peprotech, cat no.100-21) for 24 h at 5 % CO2 and 37°C prior to co-culture with T cells. HMDM were added to the culture in different ratios, e. g.1:1, 2.5:1 and 5:1 (T cell : HMDM). Total number of cells varied from 120 k to 150 k per well. Co- culture of T cells and macrophages lasted for 5 days at 37°C and 5 % CO2. As a positive control, stimulated T cells cultured without HMDM were used (max. proliferation observed). As a negative control CTV-labelled, unstimulated T cells were used (population showed no proliferation). [0347] T cell proliferation was measured by quantification of CTV dilution, which increases with every cell division. Data was collected with flow cytometry method, where T cells and HMDM were identified by staining with mAb (all from BioLegend: CD14-APC cat. no.325608, CD4-FITC 317408, CD8 PercP cat no.344708). Live/dead stain (Zombie NIR Fixable Viability Kit, BioLegend, cat. No.423106 ) was employed to exclude dead cells from subsequent analysis. Data was acquired with Attune NxT Flow Cytometer (Thermo Fisher Scientific) and analysed with the FlowJo software (Tree Star) and T cell proliferation was quantified by calculating the proliferation index, which is the total number of divisions divided by the number of cells that went into division. [0348] Trispecific molecules of the invention demonstrate a dose-dependent induction of T-cell proliferation, as indicated by the significant CTV dye dilution and proliferation index (see Figure 16). This heightened activity is attributed to the inhibition of LILRB2, as evidenced by the singular effectiveness of the IgG compound as a standalone agent. 119915P877PC 23.12.2024 Numab Therapeutics AG Magrolimab, avelumab, and atezolizumab IgGs, as well as PRO5053 and PRO5056 showed no significant impact after treatment. [0349] The induction of T cell proliferation is specifically propelled by the inhibition of LILRB2, with no discernible involvement of the PDL1- and CD47-BDs in this assay. Additionally, the trispecific antibodies of the invention exhibit dose-dependent activity in this context. 5.7 SEA Assay [0350] An SEA (Staphylococcal Enterotoxin A) assay was further performed to evaluate the potential usability of the multispecific antibodies of the invention in conditions characterized by excessive inflammation. [0351] Peripheral blood mononuclear cells (PBMC) were isolated from fresh human whole blood employing density gradient centrifugation. PBMCs were depleted of NK cells using anti-CD56 microbeads (Miltenyi Biotec, 130-097-042) and the MACS cell separation kit (Miltenyi Biotec).200,000 NK-cell depleted PBMCs per well were added to the 96-well plate (flat bottom, TPP), followed by the addition of serial dilutions of experimental compounds in assay buffer containing Staphylococcal enterotoxin A (SEA, Toxin Technology, AT 101) at a concentration of 10 ng/mL. After 76 h of incubation at 37°C and 5 % CO2, cell supernatants were harvested and human Interleukin 1β (IL-1β) and tumor necrosis cell factor alpha (TNF- α) levels in the culture supernatants were quantified using the cytometric bead array method (CBA) from LegendPlex kit provided by BioLegend according to kit instructions (cat no.: 740930). [0352] As shown in Figure 17, the inhibition of LILRB2 is responsible for mediating TNFα secretion, and the tested multispecific antibodies demonstrated comparable potency to the anti-LILRB2 IgG4 format from Numab. Treatment with magrolimab or atezolizumab did not lead to an increase in TNFα levels, and anti-PDL1 x CD47 bispecific antibodies also failed to enhance TNFα release. [0353] IL-1β induction is primarily attributed to PDL1-binding, as demonstrated by the response to atezolizumab, with no discernible impact from CD47 and LILRB2 blockade. Interestingly, the multispecific compound exhibited activity on par with the combination of three IgGs, considering data from five donors. However, it is important to note a significant variability among donors in the observed outcomes. 119915P877PC 23.12.2024 Numab Therapeutics AG 5.8 Hemagglutination assay [0354] To test whether the multispecific antibodies of the invention trigger agglutination of human erythrocytes, a hemagglutination assay based on optical density measurement was performed. The method used to assess agglutination was the same as the method described in the Example 4.2. [0355] There was no apparent hemagglutination observed for trispecific molecules PRO5052 and PRO5055 within the tested concentration range, which extended up to 500 nM, as seen for the anti-CD47 x PDL1 bispecific antibody PRO3790. Conversely, treatment with PRO2777 (anti-CD47 antibody of clone hu5F9) and PRO3599 (anti-CD47 x PDL1 bispecific antibody from Pfizer) resulted in noticeable agglutination of red blood cells (RBCs), although PRO3599 exhibited larger EC50 values than PRO2777 (Figures 18 and 19). 5.9 Cell binding to cancer cell lines [0356] To test binding of the trispecific antibodies of the invention to CD47 on the surface of cancer cells, cancer cells expressing human CD47 and PDL1 were tested in flow cytometry. The method used to determine cell-binding to cancer cell lines was the same as described in Example 4.2. [0357] The results of the binding tests are summarized in Figure 20. Biphasic binding curves were obtained on HT-1080 cells, due to sequential binding of the PDL1- and CD47-BDs to the cells. For HT-29 cells, only monophasic binding curves were obtained, as these cells do not express PDL1. PRO4672, PRO4687, PRO5052 and PRO5055 demonstrated potent binding to HT-1080 and weaker binding to HT-29 cells. The reference antibody PRO3599 exhibited a similar binding profile to both cancer cell lines, albeit with a somewhat weaker potency for HT-29 cancer cells. The reference antibodies PRO2777 and PRO3851 showed a monophasic binding profile to both cell lines with PRO3851 exhibiting the highest Emax of all tested antibodies. 5.10 Neutralization of CD47/SIRPα interaction [0358] The potency of the trispecific antibodies of the present invention to inhibit the interaction of CD47/SIRPα on NCI-H292 and CHO CD47 cells was assessed analogous to 119915P877PC 23.12.2024 Numab Therapeutics AG the procedure disclosed in Example 4.2. The results of the inhibition study are summarized in Figures 21 and 22. [0359] As apparent from these Figures, PRO5052 and PRO5055 are able to completely inhibit CD47/SIRPα interaction, with IC50 values comparable to PRO4672 and PRO4687, respectively. On the other hand, PRO3599 showed only partial SIRPα neutralization, irrespective of IFNy stimulation of NCI-H292 cells. The other reference antibodies PRO2777 (magrolimab) and PRO3851 showed similar SIRPα neutralization potencies on NCI-H292 cells. 5.11 LILRB2 selectivity assessment [0360] To assess whether the trispecific antibodies according to the invention selectively bind to LILRB2 and not to other members of protein family LILRA and LILRB, a cellular ELISA (CELISA) using transiently transfected CHO cells was developed. The method applied for this assessment was the same as described in Example 3.6. [0361] As shown in Figure 23, trispecific antibodies PRO5052 and PRO5055 demonstrated potent and selective binding to LILRB2. 5.12 Safety profile assessment in human whole blood [0362] This assay aimed to assess the safety profile of the multispecific antibodies according to the present invention in human whole blood by measuring immune cell count and activation, as well as cytokine release after treatment with test compounds. Both PRO5052 and PRO5055 multispecific antibodies were tested in parallel. Mock-compounds (hSA- binding domain) for each individual arm were included to assess possible mode of action in case of unexpected activity in human whole blood (see Table 44). The following molecules were used as positive controls: - αCD3 mAb (OKT3 clone, soluble & coated): T-cell activator, - αCD52 mAb (Alemtuzumab, soluble & coated): IgG1 based T and B cell depletion, and - PHA (Phytohemagglutinin): inducer of agglutination of human immune cells. [0363] Whole blood assay was performed according to standard procedures. The assay setup was as follows: - Donors tested: tested in 3 independent experiments with 2 human donors per replicate - Incubation conditions: 20h, 37°C, 5% CO2 119915P877PC 23.12.2024 Numab Therapeutics AG - Test compound concentration: 1, 10 and 100 μg/ml - Positive controls: PHA, αCD3, αCD52; applied in concentrations of 10 and 100 μg/ml - Read outs: 1) Immune cell count (CD45+, TCRa/b+, CD4 T cells, CD8 T cells, B cells, NK cells, CD14+ monocytes); 2) Immune cell activation (expression of CD69 & CD25 on T cells); 3) Human cytokine release: CD8/NK cell Panel (Legendplex) - IL-17A, IL-2, IL-4, IL-10, IL-6, TNF-α, Fas, FasL, IFN-γ, Granzyme A, Granzyme B, Perforin, Granulysin. [0364] The results for cytokine release of PRO5052 and PRO5055 in the whole blood assay are summarized in Table 45. PRO5052 and PRO5055 show good safety profile in human whole blood. Their overall cytokine release is minimal and magnitudes lower than positive controls PHA, αCD52, coated αCD3 and soluble αCD3. PRO5052 and PRO5055 show similar or even lower cytokine release than the clinically tested PRO3599 (Pfizer’s anti-PDL1 x CD47 mAB), which showed good safety profile in clinical studies. Results for the “Mock”- controls PRO5053, PRO5054, PRO5605, PRO5423, PRO5056, PRO5057, PRO5606 and PRO5134 were virtually identical to PRO5052 and PRO5055 and are thus not shown. [0365] The results for immune cell counts and immune cell activation are summarized in Table 46. PRO5052 and PRO5055 did not induce immune cell activation or changes in immune cell count, while the positive controls showed expected results: PHA induced depletion of human immune cells, αCD52 induced depletion of immune cells such as T cells and B cells, but not monocytes. Activation (expression of CD69) of remaining T cells in PHA and αCD52 treated samples was increased. CD25 was increased by PHA but decreased by αCD52. αCD3 treatment leads to reduced expression of TCRa/b. Results for the “Mock”- controls PRO5053, PRO5054, PRO5605, PRO5423, PRO5056, PRO5057, PRO5606 and PRO5134 were virtually identical to PRO5052 and PRO5055 and are thus not shown. Table 44: Multispecific antibodies use in whole blood assay Targets Series 1 Series 2 anti-PDL1 x CD47 x LILRB2 (Fc active) PRO5052 PRO5055 anti-PDL1 x CD47 x (mock) (Fc active) PRO5053 PRO5056 anti-PDL1 x (mock) x aLILRB2 (Fc active) PRO5054 PRO5057 anti-(mock) x CD47 x LILRB2 (Fc active) PRO5605 PRO5606 119915P877PC 23.12.2024 Numab Therapeutics AG Table 45: Induction of cytokine release Read out PRO5052 PRO5055 PRO3599 hIgG1 αCD3 αCD52 (coated) (soluble) PHA TNFα - - - - ↑ ↑ ↑↑↑ IL-6 - - - - ↑ ↑ ↑↑↑ Granzyme B - - - - - - ↑↑↑ IL-2 / IL-10 - - - - - - ↑ IFNγ / Perforin / FasL / Granulysin / No change from baseline in all conditions Granzyme A IL-4 / IL-17A / Fas No significant secretion in whole blood assay “ – “ No difference to baseline “ ↑ “ Increased from baseline level Table 46: Immune cell counts and immune cell activation Read out PRO5052 PRO5055 PRO3599 hIgG1 aCD3 aCD52 (coated) (soluble) PHA CD45 count - - - - - ↓↓ ↓↓ T cell count - - - - ↓ ↓↓ ↓↓ B cell count - - - - - ↓↓ ↓↓ CD14 count - - - - - - ↓ NK cell count - - - - - - - T cell activation / CD69 - - - - ↑ ↑ ↑↑↑ T cell activation / CD25 - - - - - ↓ ↑ “ – “ No difference to baseline “ ↑ “ Increased from baseline level “ ↓ “ Decreased from baseline level

Claims

119915P877PC 23.12.2024 Numab Therapeutics AG CLAIMS 1. A multispecific antibody comprising: a) one binding domain, which specifically binds to CD47 (CD47-BD), wherein said CD47-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequences of SEQ ID NO: 7; or ii. the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 5, and the LCDR3 sequences of SEQ ID NO: 7; b) one binding domain, which specifically binds to PDL1 (PDL1-BD); and c) an IgG-like region comprising an immunoglobulin Fc region, a first Fab arm and a second Fab arm, wherein: - the CD47 BD is an scFv fragment replacing the first Fab arm, and - the PDL1 BD is in the binding domain of the second Fab arm or is an scFv fragment replacing said second Fab arm. 2. The multispecific antibody of claim 1, wherein said CD47-BD comprises: the HCDR1 sequence of SEQ ID NO: 1, the HCDR2 sequences of SEQ ID NO: 2, the HCDR3 sequence of SEQ ID NO: 3, the LCDR1 sequence of SEQ ID NO: 4, the LCDR2 sequence of SEQ ID NO: 6, and the LCDR3 sequences of SEQ ID NO: 7. 3. The multispecific antibody of claim 1 or 2, wherein said PDL1-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, 119915P877PC 23.12.2024 Numab Therapeutics AG the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 79, and the LCDR3 sequences of SEQ ID NO: 80; or ii. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 78, and the LCDR3 sequences of SEQ ID NO: 80; iii. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 21, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequences of SEQ ID NO: 27; or iv. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 21, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequences of SEQ ID NO: 27; or v. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 26, and the LCDR3 sequences of SEQ ID NO: 27; or vi. the HCDR1 sequence of SEQ ID NO: 20, the HCDR2 sequences of SEQ ID NO: 22, the HCDR3 sequence of SEQ ID NO: 23, the LCDR1 sequence of SEQ ID NO: 24, the LCDR2 sequence of SEQ ID NO: 25, and the LCDR3 sequences of SEQ ID NO: 27; or 119915P877PC 23.12.2024 Numab Therapeutics AG particularly, wherein said PDL1-BD comprises: i. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 79, and the LCDR3 sequences of SEQ ID NO: 80; or ii. the HCDR1 sequence of SEQ ID NO: 74, the HCDR2 sequences of SEQ ID NO: 75, the HCDR3 sequence of SEQ ID NO: 76, the LCDR1 sequence of SEQ ID NO: 77, the LCDR2 sequence of SEQ ID NO: 78, and the LCDR3 sequences of SEQ ID NO: 80. 4. The multispecific antibody of any of the preceding claims, wherein the KD of the PDL1- BD for binding to human PDL1 is at least 500 fold lower, in particular at least 1000 fold lower, in particular at least 1500 fold lower, in particular at least 2000 fold lower, than the KD of the CD47 BD for binding to human CD47. 5. The multispecific antibody of any one of the preceding claims, wherein said CD47-BD comprises a) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 8 and 9; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 11 and 12, particularly to SEQ ID NO: 12; or b) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 10; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 13. 6. The multispecific antibody of any one of the preceding claims, wherein said PDL1-BD comprises a) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 28; and 119915P877PC 23.12.2024 Numab Therapeutics AG a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 34, 36 and 37, particularly selected from SEQ ID NOs: 34 and 37, particularly to SEQ ID NO:37; or b) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 29 and 31; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 33 and 35, particularly to SEQ ID NO: 35; or c) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 30; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 32 and 34, particularly to SEQ ID NO: 34; or d) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 38 and 42; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 48, 50 and 51, particularly selected from SEQ ID NOs: 48 and 51, particularly to SEQ ID NO: 51; or e) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 39, 41, 43 and 45; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 47 and 49, particularly to SEQ ID NO: 49; or f) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 40 and 44; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 46 and 48, particularly to SEQ ID NO: 48; or g) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 52; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 58, 60 and 61, particularly selected from SEQ ID NOs: 58 and 61, particularly to SEQ ID NO: 61; or h) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 53 and 55; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 57 and 59, particularly to SEQ ID NO: 59; or 119915P877PC 23.12.2024 Numab Therapeutics AG i) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 54; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 56 and 58, particularly to SEQ ID NO: 58; or j) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 81; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 86, 88 and 89, particularly selected from SEQ ID NOs: 86 and 89, particularly to SEQ ID NO: 89; or k) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 83; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 85 and 86, particularly to SEQ ID NO: 86; or l) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 82 and 84, particularly to SEQ ID NO: 84; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequences of SEQ ID NO: 86. 7. The multispecific antibody of any one of the preceding claims, wherein said multispecific antibody further comprises: d) one further binding domain, which has specificity for an antigen different from CD47 and PDL1, particularly one further binding domain, which specifically binds to LILRB2 (LILRB2-BD), wherein said one further binding domain is attached to one of the C-termini of the heavy chains of the immunoglobulin Fc region; or d) two further binding domains, which have specificity for an antigen different from CD47 and PDL1, particularly two further binding domains, which specifically bind to LILRB2 (LILRB2-BD), wherein said two further binding domains are attached to each of the C-termini of the heavy chains of the immunoglobulin Fc region. 8. The multispecific antibody of claim 7, wherein said LILRB2-BD comprises: 119915P877PC 23.12.2024 Numab Therapeutics AG i. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequences of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 98, and the LCDR3 sequences of SEQ ID NO: 99; or ii. the HCDR1 sequence of SEQ ID NO: 93, the HCDR2 sequences of SEQ ID NO: 94, the HCDR3 sequence of SEQ ID NO: 95, the LCDR1 sequence of SEQ ID NO: 96, the LCDR2 sequence of SEQ ID NO: 97, and the LCDR3 sequences of SEQ ID NO: 99; or iii. the HCDR1 sequence of SEQ ID NO: 112, the HCDR2 sequences of SEQ ID NO: 113, the HCDR3 sequence of SEQ ID NO: 114, the LCDR1 sequence of SEQ ID NO: 115, the LCDR2 sequence of SEQ ID NO: 117, and the LCDR3 sequences of SEQ ID NO: 118; or iv. the HCDR1 sequence of SEQ ID NO: 112, the HCDR2 sequences of SEQ ID NO: 113, the HCDR3 sequence of SEQ ID NO: 114, the LCDR1 sequence of SEQ ID NO: 115, the LCDR2 sequence of SEQ ID NO: 116, and the LCDR3 sequences of SEQ ID NO: 118 or v. the HCDR1 sequence of SEQ ID NO: 128, the HCDR2 sequences of SEQ ID NO: 129, the HCDR3 sequence of SEQ ID NO: 130, the LCDR1 sequence of SEQ ID NO: 131, the LCDR2 sequence of SEQ ID NO: 133, and the LCDR3 sequences of SEQ ID NO: 134; or vi. the HCDR1 sequence of SEQ ID NO: 128, the HCDR2 sequences of SEQ ID NO: 129, the HCDR3 sequence of SEQ ID NO: 130, the LCDR1 sequence of SEQ ID NO: 131, 119915P877PC 23.12.2024 Numab Therapeutics AG the LCDR2 sequence of SEQ ID NO: 132, and the LCDR3 sequences of SEQ ID NO: 134. particularly, wherein said LILRB2-BD comprises a) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 100; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 104, 106 and 107, particularly selected from SEQ ID NOs: 104 and 107, particularly to SEQ ID NO: 107; or b) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 101; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 103 and 104, particularly to SEQ ID NO: 104; or c) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 102; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 105; or d) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 119; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 122, 123 and 124, particularly selected from SEQ ID NOs: 122 and 124, particularly to SEQ ID NO: 124; or e) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 120; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 121 and 122, particularly to SEQ ID NO: 122; or f) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 135; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to any one of the amino acid sequences selected from SEQ ID NOs: 138, 139 and 140, particularly selected from SEQ ID NOs: 138 and 140, particularly to SEQ ID NO: 140; or g) a VH sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 136; and a VL sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 percent identical to the amino acid sequence of SEQ ID NO: 138. 119915P877PC 23.12.2024 Numab Therapeutics AG 9. A nucleic acid or two nucleic acids or three nucleic acids encoding the multispecific antibody of any one of claims 1 to 8. 10. A host cell or host cells comprising a vector or two vectors or three vectors comprising the nucleic acid or two nucleic acids or three nucleic acids of claim 9. 11. A pharmaceutical composition comprising the multispecific antibody of any one of claims 1 to 8 and a pharmaceutically acceptable carrier. 12. A pharmaceutical composition comprising: i. a first multispecific antibody that specifically binds to CD47 and PDL1; ii. a second antibody that specifically binds to LILRB2; and iii. a pharmaceutically acceptable carrier. 13. The pharmaceutical composition of claim 12, wherein a) the first antibody that specifically binds to CD47 and PDL1 is selected from the group consisting of: IBI-322 by Innovent Biologics Inc.; PF-07257876 by Pfizer Inc.; 6MW-3211 by Mabwell (Shanghai) Bioscience Co. Ltd.; BAT7104 by Bio-Thera Solutions, Ltd.; QL401 by QLSF Biotherapeutics, Inc.; LB-101 by Centessa Pharmaceuticals plc; Imm2520 by ImmuneOnco Biopharmaceuticals Shanghai Inc.; SHS-009 by Nanjing Sanhome Pharmaceutical Co Ltd.; BCD-106 by Biocad Biopharmaceutical Co.; TJL-1C4 by I-MAB Biopharma Co. Ltd.; SG12473 by Hangzhou Sumgen Biotech Co. Ltd.; XB014 by Exelixis Inc.; PMC-122 by PharmAbcine Inc.; ABP-160 by Abpro Corp.; papiliximab by Shaperon Inc.; JY-207 by Shenzhen Enduring Biotech Ltd.; NY2601 and NY2901 by Light Chain Bioscience AG; or the first antibody that specifically binds to CD47 and PDL1 is selected from a multispecific antibody as defined in any one of the claims 1 to 6; b) the second antibody that specifically binds to LILRB2 is selected from the group consisting of: JTX-8064 by Jounce Therapeutics Inc.; MK-4830 by Merck & Co. Inc.; IO-108 by Immune-Onc Therapeutics Inc.; NGM-707 by NGM Biopharmaceuticals Inc.; BMS-986406 by Bristol Myers Squibb Company; IOS-1002 by ImmunOs Therapeutics AG; CDX-585 by Celldex Therapeutics Inc.; SPX-303 by SparX Biopharmaceutical Corp.; CHS-1000 by Coherus BioSciences Inc.; ES009 by 119915P877PC 23.12.2024 Numab Therapeutics AG Elpiscience (Suzhou) Biopharma Ltd.; XBH41 by GV20 Therapeutics and IVS-5001 by Invectys; or the second antibody that specifically binds to LILRB2 comprises at least one LILRB2-BD as defined in claim 9. 14. The multispecific antibody of any one of claims 1 to 8 or the pharmaceutical composition of any one of claims 11 to 13 for use in the treatment of a disease, particularly a human disease, more particularly a human disease selected from cancer, more particularly a human disease selected from a benign or especially malignant tumor, solid tumors, mesothelioma, brain cancer, kidney cancer, liver cancer, adrenal gland cancer, bladder cancer, breast cancer, stomach cancer (e. g., gastric tumors), esophageal cancer, ovarian cancer, cervical cancer, thymic cancer, choriocarcinoma, oral cancer, salivary cancer, colon cancer, rectum cancer, prostate cancer, pancreatic cancer, lung cancer (e. g., non-small cell lung cancer and small cell lung cancer), vaginal cancer, thyroid cancer, melanoma (e. g., unresectable or metastatic melanoma), renal cell carcinoma, sarcoma, glioma, glioblastoma, multiple myeloma or gastrointestinal cancer, especially colon carcinoma or colorectal adenoma, a tumor of the neck and head, endometrial cancer, Cowden syndrome, Lhermitte-Duclos disease, Bannayan-Zonana syndrome, prostate hyperplasia, a neoplasia, especially of epithelial character, preferably mammary carcinoma or squamous cell carcinoma, chronic lymphocytic leukemia, chronic myelogenous leukemia (e. g., Philadelphia chromosome-positive chronic myelogenous leukemia), acute lymphoblastic leukemia (e. g., Philadelphia chromosome-positive acute lymphoblastic leukemia), non-Hodgkin’s lymphoma, plasma cell myeloma, Hodgkin’s lymphoma, a leukemia, and any combination thereof. 15. An antibody binding domain, having specificity for CD47 (CD47-BD), as defined in any one of claims 1, 2 and 5, wherein said CD47-BD is in a format selected from an Fv, a dsFv, an scFv and a dsscFv, particularly from an Fv and an scFv, particularly wherein said CD47-BD is in the form of an scFv. 16. An antibody comprising one or two, preferably one, antibody binding domains according to claim 15.
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