WO2024250809A1 - Anti-trem2 fully human antibody or antigen-binding fragment thereof, and use thereof - Google Patents
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Classifications
-
- 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
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- 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
Definitions
- the present application relates to the field of biomedicine technology, and specifically to an anti-TREM2 antibody or an antigen-binding fragment thereof and applications thereof, in particular an anti-TREM2 fully human antibody or an antigen-binding fragment thereof and applications thereof.
- Triggering receptor 2 on myeloid cells belongs to the immunoglobulin superfamily and is a transmembrane receptor.
- the physiological functions of TREM2 include cell maturation, cell proliferation, cell survival, phagocytosis, and inflammation regulation.
- TREM2 is considered to be an important pathologically induced immune signaling center and plays an important role in the activation and survival of myeloid cells.
- TREM2 has a variety of ligands, mainly free negatively charged molecules bound to the plasma membrane, including bacterial products, DNA, lipoproteins, and phospholipids.
- TREM2 After binding to the ligand, TREM2 transmits signals to the cell through DNAX activating protein 10 (DAP10) and DNAX activating protein 12 (DAP12), and then activates multiple downstream signaling pathways to produce different effects.
- DAP10 DNAX activating protein 10
- DAP12 DNAX activating protein 12
- TREM2 is a marker of macrophage infiltration in tumors. TREM2 has been found to be expressed on the surface of tumor-associated macrophages (TAMs) in many tumors and is negatively correlated with the survival of the tumor. After binding to its ligand, TREM2 can promote high expression of CD163 by TAM, amplify the inflammatory response in the local microenvironment of the tumor, and promote the occurrence and development of the tumor.
- TAM tumor-associated macrophages
- TREM2-expressing TAMs accumulate in a variety of tumors and are associated with T cell exhaustion and resistance to immune checkpoint inhibitor therapy.
- TREM2 may be an ideal target for targeting tumor myeloid infiltration and enhancing the efficacy of immune checkpoint inhibitors.
- the present application provides an anti-TREM2 antibody or an antigen-binding fragment thereof and an application thereof, wherein the anti-TREM2 antibody or an antigen-binding fragment thereof comprises heavy chain variable regions HCDR1, HCDR2, HCDR3 of complementary determining regions and/or light chain variable regions LCDR1, LCDR2, LCDR3 of complementary determining regions; the anti-TREM2 antibody or an antigen-binding fragment thereof has a strong activity of inducing ADCC effect, and can be used to enhance the immune response and/or for treating immune-related diseases of an individual such as cancer, including using the anti-TREM2 antibody or an antigen-binding fragment thereof to kill non-stimulatory myeloid cells, thereby disabling the ability of non-stimulatory myeloid cells or reducing their number.
- an anti-TREM2 antibody or an antigen-binding fragment thereof comprising:
- HCDR1 comprising the amino acid sequence of SEQ ID NO: 1, 9, 17, 25, 31, 39, 51, 59 or 67, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 1, 9, 17, 25, 31, 39, 51, 59 or 67;
- HCDR2 comprising the amino acid sequence of SEQ ID NO: 2, 10, 18, 26, 32, 52, 60 or 68, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 2, 10, 18, 26, 32, 52, 60 or 68;
- HCDR3 comprising the amino acid sequence shown in SEQ ID NO:3, 11, 19, 27, 33, 40, 53, 61 or 69, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO:3, 11, 19, 27, 33, 40, 53, 61 or 69; and/or
- LCDR2 comprising the amino acid sequence shown in SEQ ID NO:5, 13, 21, 35, 42, 47, 55 or 63, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO:5, 13, 21, 35, 42, 47, 55 or 63;
- a light chain variable region wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NOs: 8, 16, 24, 30, 38, 45, 50, 58, 66, 72 or 76, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence as shown in SEQ ID NOs: 8, 16, 24, 30, 38, 45, 50, 58, 66, 72 or 76.
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 14
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 9, 10 and 11, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% similarity to the amino acid sequences shown in SEQ ID NOs: 9, 10 and 11.
- LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 12, 13 and 14, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 12, 13 and 14;
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 19, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 14
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96% similarity to the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27.
- said LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28;
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 31, 32 and 33, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 39, 18 and 40, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 130%, 131%, 132%, 133%,
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11; and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 46, 47 and 48, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 67, 68 and 69, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96% similarity to the amino acid sequences shown in SEQ ID NOs: 67, 68 and 69.
- said LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 70, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences shown in SEQ ID NOs: 4, 5 and 70; or
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:7, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:7
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:8, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:8;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:15, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:15
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:16, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:16;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:23, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:23
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:24, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:24;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:29, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:29
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:30, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:30;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:37, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:37
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:38, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:38;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:44, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:44
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:45, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:45;
- a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO:49, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence as set forth in SEQ ID NO:49, and a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO:50, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence as set forth in SEQ ID NO:50.
- Amino acid sequence amino acid sequence as set forth in SEQ ID NO:50
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:57, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:57
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:58, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:58;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:65, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:65
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:66, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:66;
- a heavy chain variable region and a light chain variable region wherein the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71 or is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 71.
- the light chain variable region comprises the amino acid sequence of SEQ ID NO: 72 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 72; or
- (11) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 75 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 75, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 76 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 76.
- the antibody is a mouse antibody, a monkey antibody, a rabbit antibody, a chimeric antibody, a humanized antibody, or a fully human antibody; the antibody is preferably a fully human antibody.
- the anti-TREM2 antibody or its antigen-binding fragment is selected from at least one of a monoclonal antibody, a polyclonal antibody, a neutralizing antibody, an antagonistic antibody, a fucosylated antibody, a monospecific antibody, a multispecific antibody, a nanobody, a Fab fragment, a Fab' fragment, a F(ab') 2 fragment, a Fd fragment, a Fv fragment, a dAb fragment, an isolated CDR region, and a scFv.
- the anti-TREM2 antibody is a monoclonal antibody.
- the anti-TREM2 antibody or antigen-binding fragment thereof further comprises a constant region, which is a heavy chain constant region and/or a light chain constant region;
- the heavy chain constant region is the heavy chain constant region of IgG, IgA, IgM, IgE or IgD or a variant thereof;
- the heavy chain constant region is preferably the heavy chain constant region of IgG or a variant thereof, for example, the constant region of IgG1, IgG2, IgG3 or IgG4 or a variant thereof;
- the heavy chain constant region is more preferably the heavy chain constant region of IgG1 or a variant thereof; for example, the heavy chain constant region of human IgG1 or a variant thereof, whose amino acid sequence is shown in SEQ ID NO: 90 or 92.
- the light chain constant region is the constant region of a ⁇ chain or a ⁇ chain or a variant thereof, preferably the constant region of a human ⁇ chain or a variant thereof, whose amino acid sequence is shown in SEQ ID NO: 91.
- the heavy chain constant region comprises a full-length heavy chain constant region or a fragment thereof, and the fragment can be selected from a CH1 region, an Fc region, or a CH3 region.
- the heavy chain constant region or its variant comprises an IgG1 Fc region having one or more mutations that enhance antibody-dependent cell-mediated cytotoxicity (ADCC).
- ADCC antibody-dependent cell-mediated cytotoxicity
- the heavy chain constant region or its variant comprises mutations at positions 234, 235 and 243, or mutations at positions 239, 330 and 332 of the IgG1 Fc region; in some embodiments, the heavy chain constant region or its variant comprises at least one mutation at positions 234, 235, 243, 239, 330 or 332 of the IgG1 Fc region.
- the IgG1 Fc region comprises mutations of S239D, A330L, and I332E.
- the anti-TREM2 antibody or antigen-binding fragment thereof further comprises an IgG1 Fc region having at least one mutation of S239D, A330L, and I332E.
- the Fc region binds an Fc ⁇ receptor, e.g., Fc ⁇ RI, Fc ⁇ RIIa, Fc ⁇ RIIb, Fc ⁇ RIIc, Fc ⁇ RIIIa, or Fc ⁇ RIIIb.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 77, 79, 80, 82, 83, 85 or 87, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 77, 79, 80, 82, 83, 85 or 87; and/or
- a light chain comprising the amino acid sequence shown in SEQ ID NO: 78, 81, 84 or 86, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 78, 81, 84 or 86.
- the different amino acids in the amino acid sequence with at least 80% identity compared to the amino acid sequence shown in SEQ ID NO: 77, 78, 79, 80, 81, 82, 83, 84, 85, 86 or 87 are mainly or entirely located in the FR region and/or constant region of the heavy chain.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- the heavy chain comprises the amino acid sequence of SEQ ID NO:77, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:77; and the light chain comprises the amino acid sequence of SEQ ID NO:78, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:78;
- the heavy chain comprises the amino acid sequence of SEQ ID NO:79, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:79; and the light chain comprises the amino acid sequence of SEQ ID NO:78, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:78;
- the heavy chain comprises the amino acid sequence of SEQ ID NO:80, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:80; and the light chain comprises the amino acid sequence of SEQ ID NO:81, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:81;
- the heavy chain comprises the amino acid sequence of SEQ ID NO:82, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:82; and the light chain comprises the amino acid sequence of SEQ ID NO:81, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:81;
- the heavy chain comprises the amino acid sequence of SEQ ID NO:83, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:83; and the light chain comprises the amino acid sequence of SEQ ID NO:84, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:84;
- the heavy chain comprises the amino acid sequence of SEQ ID NO:85 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:85; and the light chain comprises the amino acid sequence of SEQ ID NO:86 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:86; or
- the heavy chain comprises the amino acid sequence of SEQ ID NO: 87 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 87; and the light chain comprises the amino acid sequence of SEQ ID NO: 86 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 86.
- the antibody is a multispecific antibody that specifically binds TREM2 and additionally specifically binds one or more other targets;
- the multispecific antibody such as a bispecific antibody, further comprises at least one second antibody having a second binding specificity for a second target.
- the multispecific antibody is a bispecific antibody, a trispecific antibody or a tetraspecific antibody.
- the multispecific antibody is a bispecific antibody.
- the anti-TREM2 antibody or antigen-binding fragment thereof has one or more of the following properties:
- ADCC antibody-dependent cell-mediated cytotoxicity
- CDC complement-dependent cytotoxicity
- ADCP antibody-mediated phagocytosis
- the cytokine or chemokine is selected from IFN- ⁇ , TNF- ⁇ , CXCL1, or CXCL10.
- the anti-TREM2 antibody or antigen-binding fragment thereof is capable of specifically killing, depleting or disabling TREM2+ myeloid cells; optionally, the TREM2+ myeloid cells are non-stimulatory myeloid cells, for example: CD45+, HLA-DR+, CD11c+, CD14+ and BDCA3- non-stimulatory myeloid cells; optionally, the TREM2+ myeloid cells are intratumoral myeloid cells.
- the TREM2+ cells are selected from dendritic cells, macrophages, monocytes and neutrophils; the macrophages are preferably tumor-associated macrophages (TAMs).
- TAMs tumor-associated macrophages
- the anti-TREM2 antibody or antigen-binding fragment thereof has ADCC activity.
- ADCC can occur when the antibody binds to an antigen on the surface of a pathogenic or tumorigenic target cell.
- the second aspect of the present application provides a nucleic acid molecule, which encodes the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application.
- the nucleic acid molecule includes a nucleic acid molecule encoding the heavy chain variable region and/or the light chain variable region of the anti-TREM2 antibody or antigen-binding fragment thereof described herein, respectively.
- the third aspect of the present application provides a recombinant vector, which comprises the nucleic acid molecule described in the second aspect of the present application.
- the fourth aspect of the present application provides a host cell, which comprises the nucleic acid molecule described in the second aspect of the present application or the recombinant vector described in the third aspect of the present application.
- the vector is a cloning vector; in other embodiments, the vector is an expression vector.
- the expression vector may optionally be any expression vector capable of expressing the anti-TREM2 antibody or antigen-binding fragment thereof described herein, or a moiety thereof.
- the fifth aspect of the present application provides a method for preparing an anti-TREM2 antibody or an antigen-binding fragment thereof, which comprises culturing the host cell described in the fourth aspect of the present application.
- the host cell is a prokaryotic cell. In other embodiments, the host cell is a eukaryotic cell.
- the host cell is selected from yeast cells, mammalian cells or other cells suitable for preparing antibodies or antigen-binding fragments thereof, or moieties thereof.
- Mammalian cells are, for example, Chinese hamster ovary cells (CHO), human kidney epithelial cell line 293 cells or 293T cells.
- the host cell is a recombinant CHO cell.
- the method further comprises isolating and purifying the expression products of the host cells, for example, by purification using a Protein A affinity column.
- anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application, the nucleic acid molecule described in the second aspect of the present application, the recombinant vector described in the third aspect of the present application, or the host cell described in the fourth aspect of the present application in the preparation of a drug.
- the present application relates to a method for preventing and/or treating a disease or disorder associated with TREM2, or regulating the immune response of a subject.
- the present application relates to a method for preventing and/or treating a disease or condition related to TREM2, or regulating the immune response of a subject, comprising administering to a subject in need thereof the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application, the nucleic acid molecule described in the second aspect of the present application, the recombinant vector described in the third aspect of the present application, or the host cell described in the fourth aspect of the present application.
- the sixth aspect of the present application provides the use of the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect, the nucleic acid molecule described in the second aspect of the present application, the recombinant vector described in the third aspect of the present application, or the host cell described in the fourth aspect of the present application in the preparation of a reagent for diagnosing TREM2-related diseases or conditions.
- the TREM2-related disease or condition is cancer. In some embodiments, the TREM2-related disease or condition is a lipid-related disease or condition. In some embodiments, the TREM2-related disease or condition is a fibrotic disease or condition. In some embodiments, the agent is capable of modulating an immune response in a subject.
- the agent is capable of increasing an anti-tumor immune response in a subject and/or inducing a memory immune response in a subject.
- the seventh aspect of the present application provides a kit for detecting TREM2, which comprises the anti-TREM2 antibody or its antigen-binding fragment described in the first aspect of the present application or the nucleic acid molecule described in the second aspect of the present application.
- the eighth aspect of the present application provides a method for treating or preventing a disease or condition in a subject in need thereof, comprising administering the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application to the subject.
- the disease or condition is a TREM2-related disease or condition.
- the TREM2-related disease or condition is cancer.
- the TREM2-related disease or condition is a lipid-related disease or condition.
- the TREM2-related disease or condition is a fibrotic disease or condition.
- the ninth aspect of the present application provides a method for enhancing the immune response of a subject, comprising administering the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application to the subject.
- the enhancing the subject's immune response is increasing the subject's anti-tumor immune response and/or inducing the subject's memory immune response.
- the subject is a mammal, such as a human.
- the immune response is an adaptive immune response. In some embodiments, the immune response is an innate immune response.
- the lipid-related disease or disorder is selected from obesity, fatty liver disease, heart disease, stroke, atherosclerosis, diabetes, osteoarthritis, gout, sleep apnea, or hypertension.
- the fibrotic disease or disorder is a liver disease.
- the liver disease is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).
- the monoclonal antibody targeting TREM2 in the present application blocks TREM2-mediated signal transduction or eliminates tumor-associated macrophages (TAMs) expressing TREM2 in the tumor microenvironment through at least one of the ADCC effect, CDC effect, and ADCP effect (especially through the ADCC effect), thereby helping to enhance the anti-tumor immune response.
- TAMs tumor-associated macrophages
- the anti-TREM2 antibodies or antigen-binding fragments thereof of the present application can cross-bind to human, mouse, or monkey TREM2 overexpressing cell lines or cross-bind to human or monkey TREM2 overexpressing cell lines.
- Figures 1-1 to 1-4 are graphs showing the binding test results of the fully human antibody and positive control antibody of the present application with the human TREM2 protein;
- Figures 1-5 to 1-8 are graphs showing the binding test results of the fully human antibody and positive control antibody of the present application with the mouse TREM2 protein;
- Figures 1-9 to 1-12 are graphs showing the binding test results of the fully human antibody and the positive control antibody of the present application with the cynomolgus monkey TREM2 protein;
- Figures 2-1 to 2-3 are the results of binding detection of the fully human antibody and positive control antibody of the present application with human TREM2 overexpressing cell lines;
- Figures 2-4 to 2-8 are graphs showing the binding test results of the fully human antibody and positive control antibody of the present application with the mouse TREM2 overexpressing cell line;
- Figures 2-9 to 2-10 are the results of the binding test between the fully human antibody and the positive control antibody of the present application and the cynomolgus monkey TREM2 overexpressing cell line;
- Figures 3-1 to 3-3 are ADCC effect results of the fully human antibodies of the present application.
- FIG4-1 is a graph showing the in vivo efficacy tumor growth curve of the fully human antibody of the present application.
- treatment means that after administration, the disease or its related symptoms can be inhibited, contained, alleviated, improved, mitigated, relieved or eliminated, the progression of the disease or its related symptoms can be delayed, postponed, slowed, stopped or terminated, or the recurrence of the disease or its related symptoms can be prevented, controlled or reduced.
- the term "subject” as used herein encompasses any vertebrate, e.g., mammals and non-mammals, such as humans, non-human primates, sheep, dogs, cats, horses, cows, chickens, pigs, mice, etc.
- the subject in the present application is a human.
- the term "subject” as used herein can be used interchangeably with the term "patient”.
- complementarity determining region may refer to the portion of the variable region of an antibody that confers antigen binding specificity, and may refer to an amino acid sequence found in the hypervariable region of a heavy or light chain of an immunoglobulin.
- the heavy chain may include three complementary determining regions CDR (HCDR1, HCDR2, and HCDR3; and the light chain may include three complementary determining regions CDR (LCDR1, LCDR2, and LCDR3).
- the CDRs may provide contact residues that play an important role in the binding of an antibody to its antigen or antigenic epitope.
- Fully human antibody refers to human antibodies obtained by transferring human genes encoding antibodies into genetically engineered antibody gene-deficient animals through transgenic and other technologies.
- Fully human antibody refers to human antibodies obtained by transferring human genes encoding antibodies into genetically engineered antibody gene-deficient animals through transgenic and other technologies.
- phage display technology including phage display technology, transgenic mouse technology, ribosome display technology, and RNA-peptide technology.
- an antibody When referring to an antibody, unless otherwise indicated, it is an isolated antibody.
- isolated herein means that the antibody is substantially free of other cellular components with which it is naturally associated, for example, an isolated antibody can be an antibody removed from its native or natural environment.
- the percent identity (degree of homology) between sequences can be determined by comparing two sequences, for example, using freely available computer programs commonly used for this purpose on the World Wide Web (eg www.ncbi.nlm.nih.gov), such as BLASTp or BLASTn with default settings.
- an anti-TREM2 antibody or an antigen-binding fragment thereof comprising:
- HCDR1 (heavy chain CDR1) comprising the amino acid sequence of SEQ ID NO: 1, 9, 17, 25, 31, 39, 51, 59 or 67, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 1, 9, 17, 25, 31, 39, 51, 59 or 67;
- HCDR2 (heavy chain CDR2) comprising the amino acid sequence of SEQ ID NO: 2, 10, 18, 26, 32, 52, 60 or 68, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 2, 10, 18, 26, 32, 52, 60 or 68;
- HCDR3 (heavy chain CDR3) comprising the amino acid sequence shown in SEQ ID NO: 3, 11, 19, 27, 33, 40, 53, 61 or 69, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 3, 11, 19, 27, 33, 40, 53, 61 or 69; and/or
- LCDR1 (light chain CDR1) comprises the amino acid sequence shown in SEQ ID NO: 4, 12, 20, 34, 41, 46, 54, 62 or 73, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:4, 12, 20, 34, 41, 46, 54, 62 or 73;
- LCDR2 light chain CDR2 comprising the amino acid sequence shown in SEQ ID NO: 5, 13, 21, 35, 42, 47, 55 or 63, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 5, 13, 21, 35, 42, 47, 55 or 63;
- LCDR3 light chain CDR3
- LCDR3 which comprises the amino acid sequence shown in SEQ ID NO: 6, 14, 22, 28, 36, 43, 48, 56, 64, 70 or 74, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 6, 14, 22, 28, 36, 43, 48, 56, 64, 70 or 74.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NOs: 7, 15, 23, 29, 37, 44, 49, 57, 65, 71 or 75, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence as shown in SEQ ID NOs: 7, 15, 23, 29, 37, 44, 49, 57, 65, 71 or 75; and/or
- a light chain variable region wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NOs: 8, 16, 24, 30, 38, 45, 50, 58, 66, 72 or 76, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence as shown in SEQ ID NOs: 8, 16, 24, 30, 38, 45, 50, 58, 66, 72 or 76.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 14
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 9, 10 and 11, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% similarity to the amino acid sequences shown in SEQ ID NOs: 9, 10 and 11.
- LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 12, 13 and 14, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 12, 13 and 14;
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 19, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 14
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96% similarity to the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27.
- said LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28;
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 31, 32 and 33, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 31, 32 and 33, respectively; and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 34, 35 and 36, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 39, 18 and 40, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 130%, 131%, 132%, 133%,
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 1112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 119, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%,
- HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 67, 68 and 69, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96% similarity to the amino acid sequences shown in SEQ ID NOs: 67, 68 and 69.
- said LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 70, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences shown in SEQ ID NOs: 4, 5 and 70; or
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 4, 5 and 6, respectively;
- HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 17, 18 and 19, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 20, 21 and 22, respectively; or
- HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 39, 18 and 40, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 41, 42 and 43, respectively.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 4, 5 and 6, respectively;
- HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 17, 18 and 19, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 20, 21 and 22, respectively;
- HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28, respectively; or
- HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 46, 47 and 48, respectively.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:7, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:7
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:8, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:8;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:15, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:15
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:16, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:16. amino acid sequences that are 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 95%, 95%, 97%, 98% or 99% identical to the
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:23, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:23
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:24, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:24;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:29, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:29
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:30, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:30;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:37, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:37
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:38, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:38;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:44, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:44
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:45, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:45;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:49, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:49
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:50, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:50;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:57, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:57
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:58, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:58;
- the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:65, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:65
- the light chain variable region comprises the amino acid sequence of SEQ ID NO:66, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:66;
- a heavy chain variable region and a light chain variable region wherein the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71 or is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 71.
- the light chain variable region comprises the amino acid sequence of SEQ ID NO: 72 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 72; or
- (11) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 75 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 75, and the light chain variable region comprises SEQ ID NO: NO:76 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:76.
- the different amino acids in an amino acid sequence with at least 80% identity compared to the amino acid sequence shown in SEQ ID NO: 7, 8, 15, 16, 23, 24, 29, 30, 37, 38, 44, 45, 49, 50, 57, 58, 65, 66, 71, 72, 75 or 76 are mainly or entirely located in the FR region.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- a heavy chain variable region and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 7 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 7, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 8 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 8;
- the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 23, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 24 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 24; or
- the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 44 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 44
- the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 45 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 45;
- the different amino acids in the amino acid sequence with at least 80% identity are all located in the FR region.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- a heavy chain variable region and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 7 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 7, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 8 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 8;
- the heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 23, and the light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 24 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 24;
- the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 29, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 30 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 30; or
- the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 49 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 49
- the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 50 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 50;
- the different amino acids in the amino acid sequence with at least 80% identity are all located in the FR region.
- CDR sequences numbered according to any one or more of the ImMunoGenTics (IMGT) numbering rules, the Kabat numbering rules, the Chothia numbering rules and the Contact numbering rules (based on analysis of available complex crystal structures) also fall within the scope of protection of the present application.
- IMGT ImMunoGenTics
- the anti-TREM2 antibody or its antigen-binding fragment may also be conservatively modified.
- Conservative modification refers to amino acid modification that does not significantly affect or change the binding properties of the antibody.
- a conservative amino acid substitution may be substituted with another amino acid of the same type (having similar chemical properties or functions).
- amino acids can be divided into the following categories according to their side chain properties: (1) non-polar amino acids: Ala (A), Val (V), Leu (L), Ile (I), Pro (P), Phe (F), Trp (W), Met (M); (2) uncharged polar amino acids: Gly (G), Ser (S), Thr (T), Cys (C), Tyr (Y), Asn (N), Gln (Q); (3) acidic amino acids: Asp (D), Glu (E); (4) basic amino acids: Lys (K), Arg (R), His (H).
- amino acids can be divided based on common side chain properties: (1) hydrophobic amino acids: Met, Ala, Val, Leu, Ile; (2) neutral hydrophilic amino acids: Cys, Ser, Thr, Asn, Gln; (3) acidic amino acids: Asp, Glu; (4) basic amino acids: His, Lys, Arg; (5) amino acids that affect chain orientation: Gly, Pro; (6) aromatic amino acids: Trp, Tyr, Phe.
- anti-TREM2 antibodies or antigen-binding fragments thereof and variants thereof described in the present application are all capable of specifically binding to TREM2.
- the anti-TREM2 antibody or antigen-binding fragment thereof of the present application is an isolated anti-TREM2 antibody or antigen-binding fragment thereof.
- the antibody is a mouse antibody, a monkey antibody, a rabbit antibody, a chimeric antibody, a humanized antibody, or a fully human antibody; preferably, the antibody is a fully human antibody.
- the anti-TREM2 antibody or antigen-binding fragment thereof is selected from at least one of a monoclonal antibody, a polyclonal antibody, a neutralizing antibody, an antagonistic antibody, an afucosylated antibody, a monospecific antibody, a multispecific antibody, a nanobody, a Fab fragment, a Fab' fragment, a F(ab') 2 fragment, a Fd fragment, a Fv fragment, a dAb fragment, an isolated CDR region, and a scFv.
- the anti-TREM2 antibody is a monoclonal antibody.
- the anti-TREM2 antibody or antigen-binding fragment thereof further comprises a heavy chain constant region and/or a light chain constant region;
- the heavy chain constant region is the heavy chain constant region of IgG, IgA, IgM, IgE or IgD or a variant thereof;
- the heavy chain constant region is preferably the heavy chain constant region of IgG or a variant thereof, for example, the constant region of IgG1, IgG2, IgG3 or IgG4 or a variant thereof;
- the heavy chain constant region is more preferably the heavy chain constant region of IgG1 or a variant thereof; for example, the heavy chain constant region of human IgG1 or a variant thereof, whose amino acid sequence is shown in SEQ ID NO: 90 or 92.
- the light chain constant region is the constant region of a ⁇ chain or a ⁇ chain or a variant thereof, preferably the constant region of a human ⁇ chain or a variant thereof, whose amino acid sequence is shown in SEQ ID NO: 91.
- the heavy chain constant region comprises a full-length heavy chain constant region or a fragment thereof, and the fragment can be selected from a CH1 region, an Fc region, or a CH3 region.
- the heavy chain constant region or its variant comprises an IgG1 Fc region having one or more mutations that enhance antibody-dependent cell-mediated cytotoxicity (ADCC).
- ADCC antibody-dependent cell-mediated cytotoxicity
- the heavy chain constant region or its variant comprises mutations at positions 234, 235 and 243, or mutations at positions 239, 330 and 332 of the IgG1 Fc region; in some embodiments, the heavy chain constant region or its variant comprises at least one mutation at positions 234, 235, 243, 239, 330 or 332 of the IgG1 Fc region.
- the IgG1 Fc region comprises mutations of S239D, A330L, and I332E.
- the anti-TREM2 antibody or antigen-binding fragment thereof further comprises an IgG1 Fc region having at least one mutation of S239D, A330L, and I332E.
- the Fc region binds an Fc ⁇ receptor, e.g., Fc ⁇ RI, Fc ⁇ RIIa, Fc ⁇ RIIb, Fc ⁇ RIIc, Fc ⁇ RIIIa, or Fc ⁇ RIIIb.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises the heavy chain variable region and light chain variable region described above, and an IgG1 Fc region. In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises the heavy chain variable region and light chain variable region described above, and an IgG1 Fc region and a constant region of a human ⁇ chain. In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises the heavy chain variable region and light chain variable region described above, an IgG1 Fc region, and a constant region of a human ⁇ chain and a constant region of a human ⁇ chain at a concentration ratio of 2:1.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises the heavy chain variable region and light chain variable region described above, and an IgG1 Fc region having at least one mutation of S239D, A330L, and I332E. In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises: (1) the heavy chain variable region and light chain variable region described above, (2) an IgG1 Fc region having at least one mutation among S239D, A330L and I332E; and (3) a constant region of a human ⁇ chain.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises: (1) the heavy chain variable region and light chain variable region described above, (2) an IgG1 Fc region having at least one mutation among S239D, A330L and I332E; and (3) a constant region of a human ⁇ chain and a constant region of a human ⁇ chain at a concentration ratio of 2:1.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 77, 79, 80, 82, 83, 85 or 87, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 77, 79, 80, 82, 83, 85 or 87; and/or
- a light chain comprising the amino acid sequence shown in SEQ ID NO: 78, 81, 84 or 86, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 78, 81, 84 or 86.
- the different amino acids in the amino acid sequence with at least 80% identity are mainly or entirely located in the FR region and/or constant region of the heavy chain; and/or compared with the amino acid sequence shown in SEQ ID NO: 78, 81, 84 or 86, the different amino acids in the amino acid sequence with at least 80% identity are mainly or entirely located in the FR region and/or constant region of the light chain.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- the heavy chain comprises the amino acid sequence of SEQ ID NO:77, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:77; and the light chain comprises the amino acid sequence of SEQ ID NO:78, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:78;
- the heavy chain comprises the amino acid sequence of SEQ ID NO:79, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:79; and the light chain comprises the amino acid sequence of SEQ ID NO:78, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:78;
- the heavy chain comprises the amino acid sequence of SEQ ID NO:80, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:80; and the light chain comprises the amino acid sequence of SEQ ID NO:81, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:81;
- the heavy chain comprises the amino acid sequence of SEQ ID NO:82, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:82; and the light chain comprises the amino acid sequence of SEQ ID NO:81, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:81;
- the heavy chain comprises the amino acid sequence of SEQ ID NO:83, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:83; and the light chain comprises the amino acid sequence of SEQ ID NO:84, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:84;
- the heavy chain comprises the amino acid sequence of SEQ ID NO:85 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:85; and the light chain comprises the amino acid sequence of SEQ ID NO:86 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:86; or
- the heavy chain comprises the amino acid sequence of SEQ ID NO: 87 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 87; and the light chain comprises the amino acid sequence of SEQ ID NO: 86 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 86.
- the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
- (1) a heavy chain and a light chain wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 87 or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 87; and the light chain comprises the amino acid sequence of SEQ ID NO: 86 or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 86;
- the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 77 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 77; and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 78 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 78;
- the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 83 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 83; and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 84 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 84; or
- the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 82 or an amino acid sequence that is at least 80% identical to the amino acid sequence shown in SEQ ID NO: 82; and the light chain comprises the amino acid sequence shown in SEQ ID NO: 81 or an amino acid sequence that is at least 80% identical to the amino acid sequence shown in SEQ ID NO: 81.
- the antibody is a multispecific antibody that specifically binds TREM2 and additionally specifically binds one or more other targets.
- the multispecific antibody further comprises at least one second antibody having a second binding specificity for a second target.
- the multispecific antibody is a bispecific antibody, a trispecific antibody or a tetraspecific antibody. More preferably, the multispecific antibody is a bispecific antibody.
- the anti-TREM2 antibody or antigen-binding fragment thereof has one or more of the following properties:
- ADCC antibody-dependent cell-mediated cytotoxicity
- CDC complement-dependent cytotoxicity
- ADCP antibody-mediated phagocytosis
- the one or more TREM2 ligands are selected from Escherichia coli cells, apoptotic cells, nucleic acids, anionic lipids, zwitterionic lipids, negatively charged phospholipids, phosphatidylserine, sulfatides, phosphatidylcholin, sphingomyelin, membrane phospholipids, lipidated proteins, proteolipids, lipidated peptides, and lipidated amyloid ⁇ peptide.
- the cytokine or chemokine is selected from IFN- ⁇ , TNF- ⁇ , CXCL1, or CXCL10.
- the anti-TREM2 antibody or antigen-binding fragment thereof is capable of specifically killing, depleting or disabling TREM2+ myeloid cells.
- the TREM2+ myeloid cells are non-stimulatory myeloid cells, for example: CD45+, HLA-DR+, CD11c+, CD14+ and BDCA3- non-stimulatory myeloid cells;
- the TREM2+ myeloid cells are intratumoral myeloid cells.
- the TREM2+ cells are selected from dendritic cells, macrophages, monocytes and neutrophils; the macrophages are preferably tumor-associated macrophages (TAMs).
- TAMs tumor-associated macrophages
- the anti-TREM2 antibody or antigen-binding fragment thereof has ADCC activity.
- ADCC can occur when the antibody binds to an antigen on the surface of a pathogenic or tumorigenic target cell.
- an in vitro ADCC assay such as described in U.S. Pat. No. 5,500,362 or 5,821,337 can be performed.
- Effector cells that can be used for the assay include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells.
- PBMC peripheral blood mononuclear cells
- NK natural killer cells
- the activity of the ADCC effect of the target molecule can be evaluated in vivo, for example, in an animal model such as disclosed in Clynes et al., Proc. Natl. Acad. Sci. (USA) 95: 652-656 (1998).
- the second aspect of the present application provides a nucleic acid molecule, which encodes the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application.
- the nucleic acid molecule includes a nucleic acid molecule encoding the heavy chain variable region and/or the light chain variable region of the anti-TREM2 antibody or antigen-binding fragment thereof described in the present application, respectively.
- the third aspect of the present application provides a recombinant vector, which comprises the nucleic acid molecule described in the second aspect of the present application.
- vector refers to a tool for expressing a target gene in a host cell, such as a plasmid vector, a cosmid vector, and a viral vector such as a phage vector, a lentiviral vector, an adenoviral vector, a retroviral vector, and an adeno-associated viral vector.
- the recombinant vector can be constructed by or by manipulating a plasmid (e.g., pSC101, pGV1106, pACYC177, ColE1, pKT230, pME290, pBR322, pUC8/9, pUC6, pBD9, pHC79, pIJ61, pLAFR1, pHV14, pGEX series, pET series, pUC19, etc.), a phage (e.g., ⁇ gt4 ⁇ B, ⁇ 8 Charon, ⁇ z1, M13, etc.) or a viral vector (e.g., SV40, etc.), which is commonly used in the art.
- a plasmid e.g., pSC101, pGV1106, pACYC177, ColE1, pKT230, pME290, pBR322, pUC8/9, pUC6, pBD9, pHC79, pIJ61, p
- a nucleic acid molecule in a recombinant vector, can be operably linked to a promoter.
- operably linked is intended to refer to a functional connection between a target nucleotide sequence and an expression control sequence (e.g., a promoter sequence).
- an expression control sequence e.g., a promoter sequence.
- a regulatory element can control transcription and/or translation of a target polynucleotide.
- the vector is a cloning vector; in other embodiments, the vector is an expression vector.
- the expression vector may optionally be any expression vector capable of expressing the anti-TREM2 antibody or antigen-binding fragment thereof, or a moiety thereof described herein.
- a recombinant vector can be constructed accordingly.
- the vector when constructing a vector as an expression vector for a prokaryotic host, the vector generally includes a strong promoter for transcription (e.g., pL ⁇ promoter, CMV promoter, trp promoter, lac promoter, tac promoter, T7 promoter, etc.), a ribosome binding site for initiating translation, and a transcription/translation termination sequence.
- a strong promoter for transcription e.g., pL ⁇ promoter, CMV promoter, trp promoter, lac promoter, tac promoter, T7 promoter, etc.
- an expression vector for a eukaryotic host includes a replication origin that can be operated in a eukaryotic cell, such as f1 replication origin, SV40 replication origin, pMB1 replication origin, adenovirus replication origin, AAV replication origin, and BBV replication origin, but is not limited thereto.
- the expression vector generally includes a promoter derived from a mammalian cell genome (e.g., a metallothionein promoter) or a promoter derived from a mammalian virus (e.g., adenovirus late promoter, vaccinia virus 7.5K promoter, SV40 promoter, cytomegalovirus promoter, tk promoter of HSV, etc.), and a polyadenylation sequence as a transcription termination sequence.
- Recombinant cells can be prepared by introducing a recombinant vector into a suitable host cell. Any host cell known in the art can be used in the present disclosure as long as it allows for continuous cloning and expression of the recombinant vector in a stable manner.
- prokaryotic host cells can be selected from Escherichia coli (E. coli) such as E. coli JM109, E. coli BL21, E. coli RR1, E. coli LE392, E. coli B, E. coli X1776, E. coli W3110, Bacillus species such as Bacillus subtilis (Bacillus subtilis) and Bacillus thuringiensis (Bacillus thuringiensis), and Enterobacteriaceae strains, such as Salmonella typhimurium (Salmonella typhimurium), Serratia marcescens (Serratia marcescens) and various Pseudomonas species.
- Escherichia coli E. coli
- E. coli JM109 E. coli JM109
- E. coli BL21 E. coli RR1, E. coli LE392
- E. coli B E. coli X1776
- E. coli W3110 Bac
- the eukaryotic host cell that can be used for transformation can be selected from but not limited to Saccharomyces cerevisiae (Saccharomyces cerevisiae), insect cells and animal cells, such as Sp2/0, CHO (Chinese hamster ovary) K1, CHO DG44, CHO S, CHO DXB11, CHO GS 8 KO, PER.C6, W138, BHK, COS 8 7, 293, HepG2, Huh7, 3T3, RIN, MDCK etc.
- Nucleic acid molecules or recombinant vectors carrying nucleic acid molecules can be introduced (transfected) into host cells using methods well known in the relevant art.
- the host cell when the host cell is a prokaryotic cell, CaCl2 or electroporation methods can be used to carry out this transfection.
- gene introduction can be achieved using, but not limited to, microinjection, calcium phosphate precipitation, electroporation, liposome-mediated transfection, or particle bombardment.
- the phenotype associated with the selection marker can be utilized according to methods well known in the art.
- the selection marker is a gene that confers resistance to certain antibiotics
- host cells can be grown in culture medium in the presence of antibiotics to select target transformants.
- the fourth aspect of the present application provides a host cell, which comprises the nucleic acid molecule described in the second aspect of the present application or the recombinant vector described in the third aspect of the present application.
- the host cell in the present application may be any suitable host cell for expressing an anti-TREM2 antibody or its antigen-binding fragment, or a portion thereof (moiety).
- the host cell is a prokaryotic cell.
- the host cell is a eukaryotic cell.
- the host cell is selected from yeast cells, mammalian cells, or other cells suitable for preparing antibodies or their antigen-binding fragments, or a portion thereof (moiety).
- Mammalian cells are, for example, Chinese hamster ovary cells (CHO), human kidney epithelial cell line 293 cells or 293T cells.
- the host cell is a recombinant CHO cell.
- the fifth aspect of the present application provides a method for preparing an anti-TREM2 antibody or an antigen-binding fragment thereof, which comprises culturing the host cell described in the fourth aspect of the present application.
- the method further comprises isolating and purifying the expression product of the host cell, for example, using a Protein A affinity column for purification.
- anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application, the nucleic acid molecule described in the second aspect of the present application, the recombinant vector described in the third aspect of the present application, or the host cell described in the fourth aspect of the present application in the preparation of a drug.
- the drug is a drug for preventing and/or treating a disease or condition associated with TREM2.
- the disease or condition associated with TREM2 is cancer.
- the disease or condition associated with TREM2 is a lipid-related disease or condition.
- the disease or condition associated with TREM2 is a fibrotic disease or condition.
- the drug is a drug for regulating the immune response of a subject.
- the drug is a drug for increasing the anti-tumor immune response of a subject and/or a drug for inducing a memory immune response of a subject.
- the sixth aspect of the present application provides the use of the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect, the nucleic acid molecule described in the second aspect of the present application, the recombinant vector described in the third aspect of the present application, or the host cell described in the fourth aspect of the present application in the preparation of a reagent for diagnosing TREM2-related diseases or conditions.
- the TREM2-related disease or condition is cancer. In some embodiments, the TREM2-related disease or condition is a lipid-related disease or condition. In some embodiments, the TREM2-related disease or condition is a fibrotic disease or condition. In some embodiments, the agent is capable of modulating an immune response in a subject. In some specific embodiments, the agent is capable of increasing an anti-tumor immune response in a subject and/or inducing a memory immune response in a subject.
- the seventh aspect of the present application provides a kit for detecting TREM2, which comprises the anti-TREM2 antibody or its antigen-binding fragment described in the first aspect of the present application or the nucleic acid molecule described in the second aspect of the present application.
- the eighth aspect of the present application provides a method for treating or preventing a disease or condition in a subject in need thereof, comprising administering the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application to the subject.
- the disease or condition is a TREM2-related disease or condition.
- the TREM2-related disease or condition is cancer.
- the TREM2-related disease or condition is a lipid-related disease or condition.
- the TREM2-related disease or condition is a fibrotic disease or condition.
- the ninth aspect of the present application provides a method for enhancing the immune response of a subject, comprising administering the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application to the subject.
- the enhancing the immune response of the subject is to increase the anti-tumor immune response of the subject and/or induce the memory immune response of the subject.
- the subject is a mammal, such as a human.
- the immune response is an adaptive immune response.
- the immune response is an innate immune response.
- Cancer cells that can be killed by the anti-TREM2 antibodies or antigen-binding fragments thereof of the present application include, but are not limited to, cancer cells from the following: bladder, blood, bone, bone marrow, brain, breast, colon, esophagus, gastrointestinal tract, gums, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate, skin, stomach, pancreas, testicles, tongue, cervix or uterus.
- cancer may specifically have the following histological types, although not limited to these: tumor, Malignant tumor disease.
- the cancer is selected from melanoma, renal cancer, hepatobiliary cancer, head and neck squamous cell carcinoma (HNSC), pancreatic cancer, colorectal cancer, bladder cancer, glioblastoma, prostate cancer, lung cancer, breast cancer, ovarian cancer, gastric cancer, esophageal cancer, kidney cancer, leukemia, lymphoma or mesothelioma.
- the cancer is colorectal cancer.
- the cancer is breast cancer.
- the lipid-related disease or disorder is selected from obesity, fatty liver disease, heart disease, stroke, atherosclerosis, diabetes, osteoarthritis, gout, sleep apnea, or hypertension.
- the fibrotic disease or disorder is a liver disease.
- the liver disease is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).
- the appropriate dose of the anti-TREM2 antibodies or antigen-binding fragments thereof described herein can be determined by a clinician based on factors such as the age, weight, sex, general condition, and severity of the disease of the subject; they can be administered by any conventional route known in the art, such as parenteral, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal administration.
- parenteral intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal administration.
- a single large dose can be administered, multiple divided doses can be administered over time, or the single dose can be reduced or increased in proportion to the severity of the treatment situation.
- the antibody when administered as a sustained release formulation, it can be administered at a reduced frequency.
- the TREM2 antibody PY314 from Pionyr Immunotherapeutics was used as a positive control antibody.
- the variable region amino acid sequence of PY314 is recorded in WO2019118513. It uses the IgG1 subtype and enhances its ADCC effect through the heavy chain constant region S239D/I332E/A330L mutation. Its sequence is shown below:
- PY314 heavy chain sequence SEQ NO ID: 88:
- PY314 light chain sequence SEQ NO ID: 89:
- the light chain and heavy chain genes of PY314 were synthesized and subcloned into the pcDNA3.4 expression vector, respectively, and transfected into Expi CHO cells. After culturing at 37°C and 5% CO2 , the cell culture was collected, centrifuged, and purified using a Protein A affinity column to obtain the PY314 antibody.
- Example 2 Screening of fully human recombinant antibody library
- the fully human recombinant antibody library (prepared by Sanyou Biotechnology) was screened by solid phase and liquid phase cross-screening, and the specific Fab antibodies against human, mouse, and crab-eating macaque TREM2 were enriched by trypsin elution, and the enrichment of different output sets was detected by ELISA.
- eluted phage solution was diluted 10-fold with logarithmic phase SS320 cells, incubated at 37°C for 30 min, mixed and dropped 2 ⁇ L on a plate, and cultured in a 37°C incubator overnight.
- TREM2 protein purchased from Kexia
- PBSM PBSM
- eluted phage solution was diluted 10-fold with logarithmic phase SS320 cells, incubated at 37°C for 30 min, mixed and dropped 2 ⁇ L on a plate, and cultured in a 37°C incubator overnight.
- Blocking Block with 5% PBSM at room temperature for 1 hour, and wash the plate three times with PBST.
- Termination Add 30 ⁇ L/well of TMB (purchased from SurModics) to develop color at room temperature for 5 to 10 minutes, then add 30 ⁇ L/well of stop solution (2 M sulfuric acid) to terminate the reaction, and read the data using an OD 450 microplate reader.
- TMB purchased from SurModics
- a total of 3378 clones were picked in the initial screening, and 496 positive clones with human, mouse, monkey, or human-monkey cross-binding activity at the protein ELISA level were obtained, of which 73 were molecules with unique sequences, and phage supernatants were prepared.
- Binding activity against human, mouse, and cynomolgus monkey TREM2 was detected by ELISA, of which 35 molecules had human, mouse, monkey, or human-monkey cross-binding activity. Full-length antibodies were constructed for these 35 molecules, using the IgG1 subtype. Some of the molecules have heavy chain constant region S239D/I332E/A330L mutations to enhance their ADCC effect, represented by HC, for example, A10-HC represents the A10 molecule with S239D/I332E/A330L mutations. The sequences of representative fully human antibody library molecules are shown in Table 1.
- the full-length antibody construction process is as follows: the genes of the above-mentioned 35 Fab molecules and the genes of the heavy chain and light chain constant regions ( ⁇ chain) of human IgG1 are cloned into the pcDNA3.4 expression vector respectively, and the full-length antibody expression plasmid with the correct sequence is obtained by sequencing analysis.
- the purified full-length antibody expression plasmid is transfected into Expi CHO cells, and after culturing at 37°C and 5% CO2 , the supernatant is collected by centrifugation, and the full-length antibody is purified using Protein A affinity chromatography to obtain the full-length antibody.
- Example 3 ELISA binding activity of fully human antibody library molecules
- the binding activity of the fully human antibody library molecules to human, mouse, and cynomolgus monkey TREM2 was detected by ELISA according to the following steps.
- Blocking Block with 5% PBSM at room temperature for 2 hours, and wash the plate three times with PBST.
- Figures 1-1 to 1-4 show the binding test results of the fully human antibodies and positive control antibodies of the present application with human TREM2 protein
- Figures 1-5 to 1-8 show the binding test results of the fully human antibodies and positive control antibodies of the present application with mouse TREM2 protein
- Figures 1-9 to 1-12 show the binding test results of the fully human antibodies and positive control antibodies of the present application with cynomolgus monkey TREM2 protein
- the fully human antibody library molecules can cross-bind with human, mouse, and monkey TREM2 proteins or cross-bind with human and monkey TREM2 proteins.
- Flow cytometry was used to detect the binding activity of the fully human antibody library molecules with CHO-K1 cells overexpressing human TREM2, mouse TREM2, and cynomolgus monkey TREM2 (all purchased from Yoshiman Gene), as follows.
- Figures 2-1 to 2-3 show the binding test results of the fully human antibody and positive control antibody of the present application with human TREM2 overexpressing cell lines
- Figures 2-4 to 2-8 show the binding test results of the fully human antibody and positive control antibody of the present application with mouse TREM2 overexpressing cell lines
- Figures 2-9 to 2-10 show the binding test results of the fully human antibody and positive control antibody of the present application with cynomolgus monkey TREM2 overexpressing cell lines
- Figures 2-9 to 2-10 show the binding test results of the fully human antibody and positive control antibody of the present application with cynomolgus monkey TREM2 overexpressing cell lines
- the PY314 antibody is used as a positive control
- the IgG1 isotype antibody is used as a negative control.
- the following steps were used to detect the ability of the fully human antibody library molecules to mediate ADCC effects, using human PBMC cells as effector cells and CHO-K1 cells overexpressing human TREM2 (purchased from Yoshiman Gene) as target cells.
- hu-TREM2-CHO-K1 cells were inoculated at 1 ⁇ 10 4 cells/well and 50 ⁇ L/well and incubated at 37°C for 30 min. At the same time, target cell plus effector cell wells (only target cells and PBMC cells were added), target cell non-lysis wells (only target cells were added), and target cell full lysis wells (only target cells were added) were set.
- PBMC cells prepared by Sanyou Biotechnology
- lysis rate (OD value of sample well - OD value of target cell plus effector cell well) / (OD value of target cell fully lysed well - OD value of target cell non-lysed well)
- FIGS 3-1 to 3-3 show the ADCC effect results of the fully human antibody library molecules. It can be seen that the fully human antibody library molecules A10, A10-HC, A4, A4-HC, and A11-HC have strong activity in inducing ADCC effect.
- HC represents a molecule that enhances the ADCC effect through the heavy chain constant region S239D/I332E/A330L mutation.
- TREM2 humanized C57BL/6 mice purchased from Shanghai Model Organisms
- Female TREM2 humanized mice were subcutaneously inoculated with MC38 mouse colon cancer cells at a number of 2.5E6/mouse to establish a subcutaneous tumor-bearing model.
- grouping and drug administration began according to the scheme shown in Table 2.
- the survival and general condition of the mice were observed every day, and the tumor volume and body weight were measured twice a week.
- the tumor growth curve and body weight change trend diagram to the end point of the study are shown in Figure 4-1 and Figure 4-2, respectively.
- the tumor growth inhibition rate (TGI) is shown in Table 2.
- the research results show that all molecules in the human antibody library have obvious tumor inhibition effects, among which the tumor inhibition effects of F83-HC, A4, A47, and A10-HC are significantly higher than that of PY314, and the tumor inhibition effects of A10 and A4-HC are equivalent to those of PY314. There was no significant decrease in the body weight of animals in each group during the experiment.
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Abstract
Description
本申请涉及生物医药技术领域，具体涉及一种抗TREM2抗体或其抗原结合片段及其应用，特别是抗TREM2全人源抗体或其抗原结合片段及其应用。The present application relates to the field of biomedicine technology, and specifically to an anti-TREM2 antibody or an antigen-binding fragment thereof and applications thereof, in particular an anti-TREM2 fully human antibody or an antigen-binding fragment thereof and applications thereof.
髓系细胞触发受体2(TREM2)属于免疫球蛋白超家族，是一种跨膜受体。TREM2的生理学功能包括细胞成熟、细胞增殖、细胞生存、吞噬作用、炎症调节。TREM2被认为是重要的病理诱导免疫信号中枢，在髓系细胞活化和存活中起着重要作用。TREM2有多种配体，主要是与质膜结合的游离负电荷分子，包括细菌产物、DNA、脂蛋白和磷脂。TREM2与配体结合后，通过DNAX激活蛋白10(DAP10)和DNAX激活蛋白12(DAP12)向胞内传递信号，继而激活下游多种信号通路，产生不同效应。TREM2是肿瘤中巨噬细胞浸润的标志物，已在多种肿瘤中发现TREM2在肿瘤相关巨噬细胞(TAM)表面表达，并与肿瘤的生存期呈负相关。TREM2与其配体结合后可促进TAM高表达CD163，放大肿瘤局部微环境中炎性反应，促进肿瘤的发生、发展。Triggering receptor 2 on myeloid cells (TREM2) belongs to the immunoglobulin superfamily and is a transmembrane receptor. The physiological functions of TREM2 include cell maturation, cell proliferation, cell survival, phagocytosis, and inflammation regulation. TREM2 is considered to be an important pathologically induced immune signaling center and plays an important role in the activation and survival of myeloid cells. TREM2 has a variety of ligands, mainly free negatively charged molecules bound to the plasma membrane, including bacterial products, DNA, lipoproteins, and phospholipids. After binding to the ligand, TREM2 transmits signals to the cell through DNAX activating protein 10 (DAP10) and DNAX activating protein 12 (DAP12), and then activates multiple downstream signaling pathways to produce different effects. TREM2 is a marker of macrophage infiltration in tumors. TREM2 has been found to be expressed on the surface of tumor-associated macrophages (TAMs) in many tumors and is negatively correlated with the survival of the tumor. After binding to its ligand, TREM2 can promote high expression of CD163 by TAM, amplify the inflammatory response in the local microenvironment of the tumor, and promote the occurrence and development of the tumor.
越来越多的研究发现，表达TREM2的TAM在多种肿瘤中积累，并且与T细胞耗竭以及免疫检查点抑制剂治疗耐药相关。An increasing number of studies have found that TREM2-expressing TAMs accumulate in a variety of tumors and are associated with T cell exhaustion and resistance to immune checkpoint inhibitor therapy.
因此，TREM2可能是靶向治疗肿瘤髓系浸润和增强免疫检查点抑制剂疗效的理想靶标。Therefore, TREM2 may be an ideal target for targeting tumor myeloid infiltration and enhancing the efficacy of immune checkpoint inhibitors.
发明内容Summary of the invention
针对现有技术，本申请提供一种抗TREM2抗体或其抗原结合片段及其应用，所述抗TREM2抗体或其抗原结合片段包含互补决定区的重链可变区HCDR1、HCDR2、HCDR3和/或互补决定区的轻链可变区LCDR1、LCDR2、LCDR3；所述抗TREM2抗体或其抗原结合片段具有较强的诱导ADCC效应的活性，可用于增强免疫应答和/或用于治疗个体的免疫相关疾患例如癌症，包括使用抗TREM2抗体或其抗原结合片段杀灭非刺激性髓系细胞，使非刺激性髓系细胞丧失能力或在数量上消减。In view of the prior art, the present application provides an anti-TREM2 antibody or an antigen-binding fragment thereof and an application thereof, wherein the anti-TREM2 antibody or an antigen-binding fragment thereof comprises heavy chain variable regions HCDR1, HCDR2, HCDR3 of complementary determining regions and/or light chain variable regions LCDR1, LCDR2, LCDR3 of complementary determining regions; the anti-TREM2 antibody or an antigen-binding fragment thereof has a strong activity of inducing ADCC effect, and can be used to enhance the immune response and/or for treating immune-related diseases of an individual such as cancer, including using the anti-TREM2 antibody or an antigen-binding fragment thereof to kill non-stimulatory myeloid cells, thereby disabling the ability of non-stimulatory myeloid cells or reducing their number.
本申请第一方面提供了一种抗TREM2抗体或其抗原结合片段，其包含：In a first aspect, the present application provides an anti-TREM2 antibody or an antigen-binding fragment thereof, comprising:
(a)包含下述互补决定区的重链可变区：(a) a heavy chain variable region comprising the following complementarity determining regions:
HCDR1，其包含SEQ ID NO:1、9、17、25、31、39、51、59或67所示的氨基酸序列，或与SEQ ID NO：1、9、17、25、31、39、51、59或67所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；HCDR1 comprising the amino acid sequence of SEQ ID NO: 1, 9, 17, 25, 31, 39, 51, 59 or 67, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 1, 9, 17, 25, 31, 39, 51, 59 or 67;
HCDR2，其包含SEQ ID NO:2、10、18、26、32、52、60或68所示的氨基酸序列，或与SEQ ID NO：2、10、18、26、32、52、60或68所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；HCDR2 comprising the amino acid sequence of SEQ ID NO: 2, 10, 18, 26, 32, 52, 60 or 68, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 2, 10, 18, 26, 32, 52, 60 or 68;
HCDR3，其包含SEQ ID NO:3、11、19、27、33、40、53、61或69所示的氨基酸序列，或与SEQ ID NO：3、11、19、27、33、40、53、61或69所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；和/或HCDR3 comprising the amino acid sequence shown in SEQ ID NO:3, 11, 19, 27, 33, 40, 53, 61 or 69, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO:3, 11, 19, 27, 33, 40, 53, 61 or 69; and/or
(b)包含下述互补决定区的轻链可变区：(b) a light chain variable region comprising the following complementarity determining regions:
LCDR1，其包含SEQ ID NO:4、12、20、34、41、46、54、62或73所示的氨基酸序列，或与SEQ ID NO：4、12、20、34、41、46、54、62或73所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；LCDR1 comprising the amino acid sequence shown in SEQ ID NO:4, 12, 20, 34, 41, 46, 54, 62 or 73, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO:4, 12, 20, 34, 41, 46, 54, 62 or 73;
LCDR2，其包含SEQ ID NO:5、13、21、35、42、47、55或63所示的氨基酸序列，或与SEQ ID NO：5、13、21、35、42、47、55或63所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；LCDR2 comprising the amino acid sequence shown in SEQ ID NO:5, 13, 21, 35, 42, 47, 55 or 63, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO:5, 13, 21, 35, 42, 47, 55 or 63;
LCDR3，其包含SEQ ID NO:6、14、22、28、36、43、48、56、64、70或74所示的氨基酸序列，或与SEQ ID NO：6、14、22、28、36、43、48、56、64、70或74所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。LCDR3, which comprises the amino acid sequence shown in SEQ ID NO:6, 14, 22, 28, 36, 43, 48, 56, 64, 70 or 74, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO:6, 14, 22, 28, 36, 43, 48, 56, 64, 70 or 74.
在本申请的一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：重链可变区，所述重链可变区包含SEQ ID NOs：7、15、23、29、37、44、49、57、65、71或75所示的氨基酸序列或与SEQ ID NOs：7、15、23、29、37、44、49、57、65、71或75所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的 氨基酸序列；和/或In some embodiments of the present application, the anti-TREM2 antibody or antigen-binding fragment thereof comprises: a heavy chain variable region, wherein the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NOs: 7, 15, 23, 29, 37, 44, 49, 57, 65, 71 or 75 or an amino acid sequence as shown in SEQ ID NOs: 7, 15, 23, 29, 37, 44, 49, 57, 65, 71 or 75 having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity. Amino acid sequence; and/or
轻链可变区，所述轻链可变区包含SEQ ID NOs：8、16、24、30、38、45、50、58、66、72或76所示的氨基酸序列或与SEQ ID NOs：8、16、24、30、38、45、50、58、66、72或76所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。A light chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NOs: 8, 16, 24, 30, 38, 45, 50, 58, 66, 72 or 76, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence as shown in SEQ ID NOs: 8, 16, 24, 30, 38, 45, 50, 58, 66, 72 or 76.
在本申请的一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments of the present application, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：1、2和3所示的氨基酸序列，或与SEQ ID NOs：1、2和3所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：4、5和6所示的氨基酸序列，或与SEQ ID NOs：4、5和6所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(1) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 6, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 4, 5 and 6;
(2)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：9、10和11所示的氨基酸序列，或与SEQ ID NOs：9、10和11所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：12、13和14所示的氨基酸序列，或与SEQ ID NOs：12、13和14所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(2) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 9, 10 and 11, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% similarity to the amino acid sequences shown in SEQ ID NOs: 9, 10 and 11. wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 12, 13 and 14, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 12, 13 and 14;
(3)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：17、18和19所示的氨基酸序列，或与SEQ ID NOs：17、18和19所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：20、21和22所示的氨基酸序列，或与SEQ ID NOs：20、21和22所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(3) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 19, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 20, 21 and 22, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 20, 21 and 22;
(4)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：25、26和27所示的氨基酸序列，或与SEQ ID NOs：25、26和27所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：4、5和28所示的氨基酸序列，或与SEQ ID NOs：4、5和28所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(4) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96% similarity to the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27. , 97%, 98% or 99% identical amino acid sequences; said LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28;
(5)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：31、32和33所示的氨基酸序列，或与SEQ ID NOs：31、32和33所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：34、35和36所示的氨基酸序列，或与SEQ ID NOs：34、35和36所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(5) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 31, 32 and 33, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 34, 35 and 36, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 34, 35 and 36;
(6)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：39、18和40所示的氨基酸序列，或与SEQ ID NOs：39、18和40所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：41、42和43所示的氨基酸序列，或与SEQ ID NOs：41、42和43所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(6) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 39, 18 and 40, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 130%, 131%, 132%, 133%, 137%, 138%, 139%, wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 41, 42 and 43, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 41, 42 and 43;
(7)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：17、18和11所示的氨基酸序列，或与SEQ ID NOs：17、18和11所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：46、47和48所示的氨基酸序列，或与SEQ ID NOs：46、47和48所示的氨基酸序列具有至少80％、81％、 82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(7) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11; and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 46, 47 and 48, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 46, 47 and 48, respectively. amino acid sequences that are 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical;
(8)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：51、52和53所示的氨基酸序列，或与SEQ ID NOs：51、52和53所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：54、55和56所示的氨基酸序列，或与SEQ ID NOs：54、55和56所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(8) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 51, 52 and 53, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153 wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 54, 55 and 56, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 54, 55 and 56;
(9)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：59、60和61所示的氨基酸序列，或与SEQ ID NOs：59、60和61所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：62、63和64所示的氨基酸序列，或与SEQ ID NOs：62、63和64所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(9) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 59, 60 and 61, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 62, 63 and 64, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 62, 63 and 64;
(10)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：67、68和69所示的氨基酸序列，或与SEQ ID NOs：67、68和69所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：4、5和70所示的氨基酸序列，或与SEQ ID NOs：4、5和70所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；或(10) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 67, 68 and 69, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96% similarity to the amino acid sequences shown in SEQ ID NOs: 67, 68 and 69. , 97%, 98% or 99% identical amino acid sequences; said LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 70, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences shown in SEQ ID NOs: 4, 5 and 70; or
(11)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：17、18和11所示的氨基酸序列，或与SEQ ID NOs：17、18和11所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：73、47和74所示的氨基酸序列，或与SEQ ID NOs：73、47和74所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。(11) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 1112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 119, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, The LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 73, 47 and 74, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences shown in SEQ ID NOs: 73, 47 and 74, respectively.
在本申请的一些具体实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some specific embodiments of the present application, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：1、2和3所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：4、5和6所示的氨基酸序列；或(1) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 6, respectively; or
(2)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：9、10和11所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：12、13和14所示的氨基酸序列；(2) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 9, 10 and 11, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 12, 13 and 14, respectively;
(3)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：17、18和19所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：20、21和22所示的氨基酸序列；(3) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 19, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 20, 21 and 22, respectively;
(4)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：25、26和27所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：4、5和28所示的氨基酸序列；(4) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28, respectively;
(5)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：31、32和33所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：34、35和36所示的氨基酸序列；(5) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 31, 32 and 33, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 34, 35 and 36, respectively;
(6)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：39、18和40所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：41、42和43所示的氨基酸序列；(6) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 39, 18 and 40, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 41, 42 and 43, respectively;
(7)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：17、18和11所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：46、47和48所示的氨基酸序列；(7) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 46, 47 and 48, respectively;
(8)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分 别具有SEQ ID NOs：51、52和53所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：54、55和56所示的氨基酸序列；(8) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein HCDR1, HCDR2 and HCDR3 are The LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 54, 55 and 56, respectively;
(9)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：59、60和61所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：62、63和64所示的氨基酸序列；(9) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 59, 60 and 61, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 62, 63 and 64, respectively;
(10)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：67、68和69所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：4、5和70所示的氨基酸序列；或(10) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 67, 68 and 69, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 70, respectively; or
(11)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：17、18和11所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：73、47和74所示的氨基酸序列。(11) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 73, 47 and 74, respectively.
在本申请的一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments of the present application, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：7所示的氨基酸序列或与SEQ ID NO：7所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：8所示的氨基酸序列或与SEQ ID NO：8所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(1) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:7, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:7, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:8, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:8;
(2)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：15所示的氨基酸序列或与SEQ ID NO：15所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：16所示的氨基酸序列或与SEQ ID NO：16所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(2) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:15, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:15, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:16, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:16;
(3)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：23所示的氨基酸序列或与SEQ ID NO：23所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：24所示的氨基酸序列或与SEQ ID NO：24所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(3) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:23, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:23, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:24, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:24;
(4)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：29所示的氨基酸序列或与SEQ ID NO：29所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：30所示的氨基酸序列或与SEQ ID NO：30所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(4) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:29, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:29, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:30, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:30;
(5)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：37所示的氨基酸序列或与SEQ ID NO：37所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：38所示的氨基酸序列或与SEQ ID NO：38所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(5) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:37, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:37, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:38, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:38;
(6)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：44所示的氨基酸序列或与SEQ ID NO：44所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：45所示的氨基酸序列或与SEQ ID NO：45所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(6) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:44, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:44, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:45, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:45;
(7)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：49所示的氨基酸序列或与SEQ ID NO：49所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：50所示的氨基酸序列或与SEQ ID NO：50所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的 氨基酸序列；(7) a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO:49, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence as set forth in SEQ ID NO:49, and a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO:50, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence as set forth in SEQ ID NO:50. Amino acid sequence;
(8)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：57所示的氨基酸序列或与SEQ ID NO：57所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：58所示的氨基酸序列或与SEQ ID NO：58所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(8) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:57, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:57, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:58, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:58;
(9)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：65所示的氨基酸序列或与SEQ ID NO：65所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：66所示的氨基酸序列或与SEQ ID NO：66所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(9) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:65, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:65, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:66, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:66;
(10)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：71所示的氨基酸序列或与SEQ ID NO：71所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：72所示的氨基酸序列或与SEQ ID NO：72所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；或(10) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71 or is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 71. The light chain variable region comprises the amino acid sequence of SEQ ID NO: 72 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 72; or
(11)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：75所示的氨基酸序列或与SEQ ID NO：75所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：76所示的氨基酸序列或与SEQ ID NO：76所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。(11) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 75 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 75, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 76 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 76.
在本申请的一些实施方式中，所述抗体为鼠源抗体、猴源抗体、兔源抗体、嵌合抗体、人源化抗体、全人源抗体；所述抗体优选为全人源抗体。In some embodiments of the present application, the antibody is a mouse antibody, a monkey antibody, a rabbit antibody, a chimeric antibody, a humanized antibody, or a fully human antibody; the antibody is preferably a fully human antibody.
在本申请的一些实施方式中，所述抗TREM2抗体或其抗原结合片段选自单克隆抗体、多克隆抗体、中和抗体、拮抗性抗体、无海藻糖基化抗体、单特异性抗体、多特异性抗体、纳米抗体、Fab片段、Fab'片段、F(ab')2片段、Fd片段、Fv片段、dAb片段、分离的CDR区和scFv中至少一种。In some embodiments of the present application, the anti-TREM2 antibody or its antigen-binding fragment is selected from at least one of a monoclonal antibody, a polyclonal antibody, a neutralizing antibody, an antagonistic antibody, a fucosylated antibody, a monospecific antibody, a multispecific antibody, a nanobody, a Fab fragment, a Fab' fragment, a F(ab') 2 fragment, a Fd fragment, a Fv fragment, a dAb fragment, an isolated CDR region, and a scFv.
在本申请的一些实施方式中，所述抗TREM2抗体为单克隆抗体。In some embodiments of the present application, the anti-TREM2 antibody is a monoclonal antibody.
在本申请的一些实施方式中，所述抗TREM2抗体或其抗原结合片段进一步包含恒定区，所述恒定区为重链恒定区和/或轻链恒定区；In some embodiments of the present application, the anti-TREM2 antibody or antigen-binding fragment thereof further comprises a constant region, which is a heavy chain constant region and/or a light chain constant region;
优选地，所述重链恒定区为IgG、IgA、IgM、IgE或IgD的重链恒定区或其变体；所述重链恒定区优选为IgG的重链恒定区或其变体，例如IgG1、IgG2、IgG3或IgG4的恒定区或其变体；所述重链恒定区更优选为IgG1的重链恒定区或其变体；例如人IgG1的重链恒定区或其变体，其氨基酸序列如SEQ ID NO：90或92所示。Preferably, the heavy chain constant region is the heavy chain constant region of IgG, IgA, IgM, IgE or IgD or a variant thereof; the heavy chain constant region is preferably the heavy chain constant region of IgG or a variant thereof, for example, the constant region of IgG1, IgG2, IgG3 or IgG4 or a variant thereof; the heavy chain constant region is more preferably the heavy chain constant region of IgG1 or a variant thereof; for example, the heavy chain constant region of human IgG1 or a variant thereof, whose amino acid sequence is shown in SEQ ID NO: 90 or 92.
优选地，轻链恒定区为κ链或λ链的恒定区或其变体，优选人κ链的恒定区或其变体，其氨基酸序列如SEQ ID NO：91所示。Preferably, the light chain constant region is the constant region of a κ chain or a λ chain or a variant thereof, preferably the constant region of a human κ chain or a variant thereof, whose amino acid sequence is shown in SEQ ID NO: 91.
在一些实施方式中，所述重链恒定区包含全长重链恒定区或其片段，所述片段可选自CH1区、Fc区或CH3区。In some embodiments, the heavy chain constant region comprises a full-length heavy chain constant region or a fragment thereof, and the fragment can be selected from a CH1 region, an Fc region, or a CH3 region.
在一些实施方式中，所述重链恒定区或其变体包含具有一种或多种增强抗体依赖性细胞介导的细胞毒性(ADCC)的突变的IgG1 Fc区。In some embodiments, the heavy chain constant region or its variant comprises an IgG1 Fc region having one or more mutations that enhance antibody-dependent cell-mediated cytotoxicity (ADCC).
在一些实施方式中，所述重链恒定区或其变体包含IgG1 Fc区第234位、第235位和第243位突变，或第239、第330和第332位突变；在一些实施方式中，所述重链恒定区或其变体包含IgG1 Fc区第234位、第235位、第243位、第239、第330或第332位的至少一种突变。In some embodiments, the heavy chain constant region or its variant comprises mutations at positions 234, 235 and 243, or mutations at positions 239, 330 and 332 of the IgG1 Fc region; in some embodiments, the heavy chain constant region or its variant comprises at least one mutation at positions 234, 235, 243, 239, 330 or 332 of the IgG1 Fc region.
在本申请的一些具体实施方式中，IgG1 Fc区包含S239D、A330L和I332E的突变。在一些实施方式中，所述抗TREM2抗体或其抗原结合片段进一步包含具有S239D、A330L和I332E中的至少一种突变的IgG1 Fc区。In some specific embodiments of the present application, the IgG1 Fc region comprises mutations of S239D, A330L, and I332E. In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof further comprises an IgG1 Fc region having at least one mutation of S239D, A330L, and I332E.
在一些实施方案中，Fc区结合Fcγ受体，例如：FcγRI、FcγRIIa、FcγRIIb、FcγRIIc、FcγRIIIa或FcγRIIIb。 In some embodiments, the Fc region binds an Fcγ receptor, e.g., FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, or FcγRIIIb.
在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
重链，所述重链包含SEQ ID NO：77、79、80、82、83、85或87所示的氨基酸序列或与SEQ ID NO：77、79、80、82、83、85或87所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；和/或A heavy chain comprising the amino acid sequence shown in SEQ ID NO: 77, 79, 80, 82, 83, 85 or 87, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 77, 79, 80, 82, 83, 85 or 87; and/or
轻链，所述轻链包含SEQ ID NO：78、81、84或86所示的氨基酸序列或与SEQ ID NO：78、81、84或86所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。A light chain comprising the amino acid sequence shown in SEQ ID NO: 78, 81, 84 or 86, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 78, 81, 84 or 86.
在一些实施方式中，与SEQ ID NO：77、78、79、80、81、82、83、84、85、86或87所示的氨基酸序列相比，具有至少80％同一性的氨基酸序列中的差异的氨基酸主要或全部位于所述重链的FR区和/或恒定区中。In some embodiments, the different amino acids in the amino acid sequence with at least 80% identity compared to the amino acid sequence shown in SEQ ID NO: 77, 78, 79, 80, 81, 82, 83, 84, 85, 86 or 87 are mainly or entirely located in the FR region and/or constant region of the heavy chain.
在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)重链和轻链，所述重链包含SEQ ID NO：77所示的氨基酸序列或与SEQ ID NO：77所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：78所示的氨基酸序列或与SEQ ID NO：78所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(1) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:77, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:77; and the light chain comprises the amino acid sequence of SEQ ID NO:78, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:78;
(2)重链和轻链，所述重链包含SEQ ID NO：79所示的氨基酸序列或与SEQ ID NO：79所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：78所示的氨基酸序列或与SEQ ID NO：78所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(2) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:79, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:79; and the light chain comprises the amino acid sequence of SEQ ID NO:78, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:78;
(3)重链和轻链，所述重链包含SEQ ID NO：80所示的氨基酸序列或与SEQ ID NO：80所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：81所示的氨基酸序列或与SEQ ID NO：81所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(3) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:80, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:80; and the light chain comprises the amino acid sequence of SEQ ID NO:81, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:81;
(4)重链和轻链，所述重链包含SEQ ID NO：82所示的氨基酸序列或与SEQ ID NO：82所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：81所示的氨基酸序列或与SEQ ID NO：81所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(4) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:82, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:82; and the light chain comprises the amino acid sequence of SEQ ID NO:81, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:81;
(5)重链和轻链，所述重链包含SEQ ID NO：83所示的氨基酸序列或与SEQ ID NO：83所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：84所示的氨基酸序列或与SEQ ID NO：84所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(5) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:83, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:83; and the light chain comprises the amino acid sequence of SEQ ID NO:84, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:84;
(6)重链和轻链，所述重链包含SEQ ID NO：85所示的氨基酸序列或与SEQ ID NO：85所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：86所示的氨基酸序列或与SEQ ID NO：86所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；或者(6) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:85 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:85; and the light chain comprises the amino acid sequence of SEQ ID NO:86 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:86; or
(7)重链和轻链，所述重链包含SEQ ID NO：87所示的氨基酸序列或与SEQ ID NO：87所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：86所示的氨基酸序列或与SEQ ID NO：86所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。(7) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 87 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 87; and the light chain comprises the amino acid sequence of SEQ ID NO: 86 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 86.
在一些实施方式中，所述抗体为多特异性抗体，所述多特异性抗体特异性结合TREM2，并且额外地特异性结合一个或多个其他靶标；In some embodiments, the antibody is a multispecific antibody that specifically binds TREM2 and additionally specifically binds one or more other targets;
优选地，所述多特异性抗体、例如双特异性抗体还包含至少一种具有针对第二靶标的第二结合特异性的第二抗体。Preferably, the multispecific antibody, such as a bispecific antibody, further comprises at least one second antibody having a second binding specificity for a second target.
优选地，所述多特异性抗体为双特异性抗体、三特异性抗体或四特异性抗体。 Preferably, the multispecific antibody is a bispecific antibody, a trispecific antibody or a tetraspecific antibody.
更优选地，所述多特异性抗体为双特异性抗体。More preferably, the multispecific antibody is a bispecific antibody.
作为非限制性示例，所述抗TREM2抗体或其抗原结合片段具有以下一个或多个特性：As non-limiting examples, the anti-TREM2 antibody or antigen-binding fragment thereof has one or more of the following properties:
(1)能够结合至人TREM2；(1) Ability to bind to human TREM2;
(2)能够结合至鼠TREM2；(2) able to bind to mouse TREM2;
(3)能够结合至食蟹猴TREM2；(3) able to bind to cynomolgus monkey TREM2;
(4)与一种或多种TREM2配体竞争结合TREM2；(4) competing with one or more TREM2 ligands for binding to TREM2;
(5)抑制和/或阻断TREM2与配体的结合；(5) inhibiting and/or blocking the binding of TREM2 to ligands;
(6)抑制和/或阻断由TREM2结合配体介导的细胞内信号传导；(6) inhibiting and/or blocking intracellular signal transduction mediated by TREM2 binding ligand;
(7)诱导细胞因子或趋化因子分泌；(7) Induce cytokine or chemokine secretion;
(8)具有抗体依赖性细胞介导的细胞毒性(ADCC)活性、补体依赖性细胞毒性(CDC)活性和抗体介导的吞噬作用(ADCP)活性中的至少一种；(8) having at least one of antibody-dependent cell-mediated cytotoxicity (ADCC) activity, complement-dependent cytotoxicity (CDC) activity, and antibody-mediated phagocytosis (ADCP) activity;
(9)增强免疫应答；(9) Enhance immune response;
(10)增强的ADCC活性。(10) Enhanced ADCC activity.
在一些实施方案中，所述一种或多种TREM2配体选自大肠杆菌细胞、凋亡细胞、核酸、阴离子性脂质、两性离子性脂质、带负电的磷脂、磷脂酰丝氨酸、硫苷脂、磷脂酰胆碱(phosphatidylcholine)、鞘磷脂、膜磷脂、脂化蛋白、蛋白脂质、脂化肽及脂化β淀粉样肽。In some embodiments, the one or more TREM2 ligands are selected from Escherichia coli cells, apoptotic cells, nucleic acids, anionic lipids, zwitterionic lipids, negatively charged phospholipids, phosphatidylserine, sulfatides, phosphatidylcholine, sphingomyelin, membrane phospholipids, lipidated proteins, proteolipids, lipidated peptides, and lipidated amyloid β peptide.
在一些实施方案中，所述细胞因子或趋化因子选自IFN-γ、TNF-α、CXCL1或CXCL10。In some embodiments, the cytokine or chemokine is selected from IFN-γ, TNF-α, CXCL1, or CXCL10.
在一些实施方案中，所述抗TREM2抗体或其抗原结合片段能够对TREM2+髓系细胞进行特异性杀灭、消减或使其丧失能力；任选地，所述TREM2+髓系细胞为非刺激性髓系细胞，例如：CD45+、HLA-DR+、CD11c+、CD14+和BDCA3-非刺激性髓系细胞；任选地，所述TREM2+髓系细胞为肿瘤内髓系细胞。In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof is capable of specifically killing, depleting or disabling TREM2+ myeloid cells; optionally, the TREM2+ myeloid cells are non-stimulatory myeloid cells, for example: CD45+, HLA-DR+, CD11c+, CD14+ and BDCA3- non-stimulatory myeloid cells; optionally, the TREM2+ myeloid cells are intratumoral myeloid cells.
任选地，所述TREM2+细胞选自树突细胞、巨噬细胞、单核细胞和嗜中性白细胞；所述巨噬细胞优选为肿瘤相关巨噬细胞(TAM)。Optionally, the TREM2+ cells are selected from dendritic cells, macrophages, monocytes and neutrophils; the macrophages are preferably tumor-associated macrophages (TAMs).
在一些具体实施方案中，所述抗TREM2抗体或其抗原结合片段具有ADCC活性。ADCC可在抗体结合病原性或致肿瘤性靶标细胞的表面上的抗原时发生。In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof has ADCC activity. ADCC can occur when the antibody binds to an antigen on the surface of a pathogenic or tumorigenic target cell.
本申请第二方面提供了一种核酸分子，所述核酸分子编码本申请第一方面所述的抗TREM2抗体或其抗原结合片段。The second aspect of the present application provides a nucleic acid molecule, which encodes the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application.
在一些实施方式中，所述核酸分子包括分别编码本文所述的抗TREM2抗体或其抗原结合片段的重链可变区或/和轻链可变区的核酸分子。In some embodiments, the nucleic acid molecule includes a nucleic acid molecule encoding the heavy chain variable region and/or the light chain variable region of the anti-TREM2 antibody or antigen-binding fragment thereof described herein, respectively.
本申请第三方面提供了重组载体，其包含本申请第二方面所述的核酸分子。The third aspect of the present application provides a recombinant vector, which comprises the nucleic acid molecule described in the second aspect of the present application.
本申请第四方面提供了宿主细胞，其包含本申请第二方面所述的核酸分子或本申请第三方面所述的重组载体。The fourth aspect of the present application provides a host cell, which comprises the nucleic acid molecule described in the second aspect of the present application or the recombinant vector described in the third aspect of the present application.
在一些实施方式中，所述载体为克隆载体；在另一些实施方式中，所述载体为表达载体。所述表达载体可任选地为能够表达本文所述抗TREM2抗体或其抗原结合片段、或其部分(moiety)的任意表达载体。In some embodiments, the vector is a cloning vector; in other embodiments, the vector is an expression vector. The expression vector may optionally be any expression vector capable of expressing the anti-TREM2 antibody or antigen-binding fragment thereof described herein, or a moiety thereof.
本申请第五方面提供了制备抗TREM2抗体或其抗原结合片段的方法，其包括培养本申请第四方面所述的宿主细胞。The fifth aspect of the present application provides a method for preparing an anti-TREM2 antibody or an antigen-binding fragment thereof, which comprises culturing the host cell described in the fourth aspect of the present application.
在一些实施方式中，所述宿主细胞为原核细胞。在另一些实施方式中，所述宿主细胞为真核细胞。In some embodiments, the host cell is a prokaryotic cell. In other embodiments, the host cell is a eukaryotic cell.
在一些实施方式中，所述宿主细胞选自酵母细胞、哺乳动物细胞或适用于制备抗体或其抗原结合片段、或其部分(moiety)的其他细胞。哺乳动物细胞例如为中国仓鼠卵巢细胞(CHO)、人肾上皮细胞系293细胞或293T细胞。在一些实施方式中，所述宿主细胞为重组CHO细胞。In some embodiments, the host cell is selected from yeast cells, mammalian cells or other cells suitable for preparing antibodies or antigen-binding fragments thereof, or moieties thereof. Mammalian cells are, for example, Chinese hamster ovary cells (CHO), human kidney epithelial cell line 293 cells or 293T cells. In some embodiments, the host cell is a recombinant CHO cell.
在一些实施方式中，所述方法还包括对所述宿主细胞的表达产物进行分离和纯化，例如采用Protein A亲和柱纯化。In some embodiments, the method further comprises isolating and purifying the expression products of the host cells, for example, by purification using a Protein A affinity column.
本申请第一方面所述的抗TREM2抗体或其抗原结合片段、本申请第二方面所述的核酸分子、本申请第三方面所述的重组载体或本申请第四方面所述的宿主细胞在制备药物中的用途。The use of the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application, the nucleic acid molecule described in the second aspect of the present application, the recombinant vector described in the third aspect of the present application, or the host cell described in the fourth aspect of the present application in the preparation of a drug.
在一些实施方式中，所述药物为预防和/或治疗TREM2相关的疾病或病症的药物。在一些实施方式中，所述TREM2相关的疾病或病症为癌症。在一些实施方式中，所述TREM2相关的疾病或病症为脂质相关疾病或病症。在一些实施方式中，所述TREM2相关的疾病或病症为纤维化疾病或病症。在一些实施方式中，所述药物为调节受试者的免疫应答的药物。在一些具体实施方式中，所述药物为增加受试者的抗肿瘤免疫应答的药物和/或诱导受试者的记忆免疫应答的药物。In some embodiments, the drug is a drug for preventing and/or treating a disease or condition associated with TREM2. In some embodiments, the disease or condition associated with TREM2 is cancer. In some embodiments, the disease or condition associated with TREM2 is a lipid-related disease or condition. In some embodiments, the disease or condition associated with TREM2 is a fibrotic disease or condition. In some embodiments, the drug is a drug for regulating the immune response of a subject. In some specific embodiments, the drug is a drug for increasing the anti-tumor immune response of a subject and/or a drug for inducing a memory immune response of a subject.
在一些实施方式中，本申请涉及用于预防和/或治疗TREM2相关的疾病或病症、或者调节受试者的免 疫应答的本申请第一方面所述的抗TREM2抗体或其抗原结合片段、本申请第二方面所述的核酸分子、本申请第三方面所述的重组载体或本申请第四方面所述的宿主细胞。或者，本申请涉及一种预防和/或治疗TREM2相关的疾病或病症、或者调节受试者的免疫应答的方法，包括向有需要的受试者给予本申请第一方面所述的抗TREM2抗体或其抗原结合片段、本申请第二方面所述的核酸分子、本申请第三方面所述的重组载体或本申请第四方面所述的宿主细胞。In some embodiments, the present application relates to a method for preventing and/or treating a disease or disorder associated with TREM2, or regulating the immune response of a subject. Alternatively, the present application relates to a method for preventing and/or treating a disease or condition related to TREM2, or regulating the immune response of a subject, comprising administering to a subject in need thereof the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application, the nucleic acid molecule described in the second aspect of the present application, the recombinant vector described in the third aspect of the present application, or the host cell described in the fourth aspect of the present application.
本申请第六方面提供了第一方面所述的抗TREM2抗体或其抗原结合片段、本申请第二方面所述的核酸分子、本申请第三方面所述的重组载体或本申请第四方面所述的宿主细胞胞在制备诊断TREM2相关的疾病或病症的试剂中的用途。The sixth aspect of the present application provides the use of the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect, the nucleic acid molecule described in the second aspect of the present application, the recombinant vector described in the third aspect of the present application, or the host cell described in the fourth aspect of the present application in the preparation of a reagent for diagnosing TREM2-related diseases or conditions.
在一些实施方式中，所述TREM2相关的疾病或病症为癌症。在一些实施方式中，所述TREM2相关的疾病或病症为脂质相关疾病或病症。在一些实施方式中，所述TREM2相关的疾病或病症为纤维化疾病或病症。在一些实施方式中，所述试剂能够调节受试者的免疫应答。In some embodiments, the TREM2-related disease or condition is cancer. In some embodiments, the TREM2-related disease or condition is a lipid-related disease or condition. In some embodiments, the TREM2-related disease or condition is a fibrotic disease or condition. In some embodiments, the agent is capable of modulating an immune response in a subject.
在一些具体实施方式中，所述试剂能够增加受试者抗肿瘤免疫应答和/或诱导受试者的记忆免疫应答。In some embodiments, the agent is capable of increasing an anti-tumor immune response in a subject and/or inducing a memory immune response in a subject.
本申请第七方面提供了一种用于检测TREM2的试剂盒，其包含本申请第一方面所述的抗TREM2抗体或其抗原结合片段或本申请第二方面所述的核酸分子。The seventh aspect of the present application provides a kit for detecting TREM2, which comprises the anti-TREM2 antibody or its antigen-binding fragment described in the first aspect of the present application or the nucleic acid molecule described in the second aspect of the present application.
本申请第八方面提供了一种治疗或预防有需要的受试者的疾病或病症的方法，其包括向所述受试者施用本申请第一方面所述的抗TREM2抗体或其抗原结合片段。The eighth aspect of the present application provides a method for treating or preventing a disease or condition in a subject in need thereof, comprising administering the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application to the subject.
在一些实施方式中，所述疾病或病症为TREM2相关的疾病或病症。在一些实施方式中，所述TREM2相关的疾病或病症为癌症。在一些实施方式中，所述TREM2相关的疾病或病症为脂质相关疾病或病症。在一些实施方式中，所述TREM2相关的疾病或病症为纤维化疾病或病症。In some embodiments, the disease or condition is a TREM2-related disease or condition. In some embodiments, the TREM2-related disease or condition is cancer. In some embodiments, the TREM2-related disease or condition is a lipid-related disease or condition. In some embodiments, the TREM2-related disease or condition is a fibrotic disease or condition.
本申请第九方面提供了一种增强受试者的免疫应答的方法，其包括向所述受试者施用本申请第一方面所述的抗TREM2抗体或其抗原结合片段。The ninth aspect of the present application provides a method for enhancing the immune response of a subject, comprising administering the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application to the subject.
优选地，所述增强受试者的免疫应答为增加受试者的抗肿瘤免疫应答和/或诱导受试者的记忆免疫应答。Preferably, the enhancing the subject's immune response is increasing the subject's anti-tumor immune response and/or inducing the subject's memory immune response.
在一些具体的实施方式中，所述受试者为哺乳动物，例如人。In some specific embodiments, the subject is a mammal, such as a human.
在一些实施方式中，所述免疫应答是适应性免疫应答。在一些实施方式中，所述免疫应答是先天性免疫应答。In some embodiments, the immune response is an adaptive immune response. In some embodiments, the immune response is an innate immune response.
在一些实施方式中，所述癌症选自黑素瘤、肾癌、肝胆癌、头颈部鳞状癌(HNSC)、胰腺癌、结直肠癌、膀胱癌、胶质母细胞瘤、前列腺癌、肺癌、乳腺癌、卵巢癌、胃癌、食道癌、肾脏癌、白血病、淋巴瘤或间皮瘤。在一些具体实施方式中，所述癌症为结直肠癌。在一些具体实施方式中，所述癌症为乳腺癌。In some embodiments, the cancer is selected from melanoma, renal cancer, hepatobiliary cancer, head and neck squamous cell carcinoma (HNSC), pancreatic cancer, colorectal cancer, bladder cancer, glioblastoma, prostate cancer, lung cancer, breast cancer, ovarian cancer, gastric cancer, esophageal cancer, kidney cancer, leukemia, lymphoma or mesothelioma. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is breast cancer.
在一些实施方式中，所述脂质相关疾病或病症选自肥胖症、脂肪肝病、心脏病、中风、动脉粥样硬化、糖尿病、骨关节炎、痛风、睡眠呼吸暂停或高血压。In some embodiments, the lipid-related disease or disorder is selected from obesity, fatty liver disease, heart disease, stroke, atherosclerosis, diabetes, osteoarthritis, gout, sleep apnea, or hypertension.
在一些实施方式中，所述纤维化疾病或病症是肝病。任选地，所述肝病为非酒精性脂肪性肝病(NAFLD)或非酒精性脂肪性肝炎(NASH)。In some embodiments, the fibrotic disease or disorder is a liver disease. Optionally, the liver disease is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).
本申请的有益效果：Beneficial effects of this application:
本申请的抗TREM2抗体或其抗原结合片段具有抗体依赖的细胞毒性(ADCC)活性、补体依赖性细胞毒性(CDC)活性、抗体介导的吞噬作用(ADCP)活性、结合亲和力，尤其具有较强的诱导ADCC效应的活性，可用于增强免疫应答和/或用于治疗个体的免疫相关疾患例如癌症，包括使用抗TREM2抗体或其抗原结合片段杀灭非刺激性髓系细胞，使非刺激性髓系细胞丧失能力或在数量上消减。The anti-TREM2 antibodies or antigen-binding fragments thereof of the present application have antibody-dependent cellular cytotoxicity (ADCC) activity, complement-dependent cytotoxicity (CDC) activity, antibody-mediated phagocytosis (ADCP) activity, binding affinity, and particularly have a strong activity of inducing ADCC effect, and can be used to enhance immune response and/or for treating individual immune-related diseases such as cancer, including using anti-TREM2 antibodies or antigen-binding fragments thereof to kill non-stimulatory myeloid cells, thereby disabling the ability of non-stimulatory myeloid cells or reducing their number.
本申请靶向TREM2的单克隆抗体阻断TREM2介导的信号转导或通过ADCC效应、CDC效应、ADCP效应中至少一种(尤其是通过ADCC效应)，清除肿瘤微环境中表达TREM2的肿瘤相关巨噬细胞(TAM)，有助于增强抗肿瘤免疫应答。The monoclonal antibody targeting TREM2 in the present application blocks TREM2-mediated signal transduction or eliminates tumor-associated macrophages (TAMs) expressing TREM2 in the tumor microenvironment through at least one of the ADCC effect, CDC effect, and ADCP effect (especially through the ADCC effect), thereby helping to enhance the anti-tumor immune response.
本申请的抗TREM2抗体或其抗原结合片段能够与人、鼠、猴TREM2过表达细胞株交叉结合或与人、猴TREM2过表达细胞株交叉结合。The anti-TREM2 antibodies or antigen-binding fragments thereof of the present application can cross-bind to human, mouse, or monkey TREM2 overexpressing cell lines or cross-bind to human or monkey TREM2 overexpressing cell lines.
图1-1～图1-4为本申请全人抗体和阳性对照抗体与人TREM2蛋白的结合检测结果图；Figures 1-1 to 1-4 are graphs showing the binding test results of the fully human antibody and positive control antibody of the present application with the human TREM2 protein;
图1-5～图1-8为本申请全人抗体和阳性对照抗体与小鼠TREM2蛋白的结合检测结果图；Figures 1-5 to 1-8 are graphs showing the binding test results of the fully human antibody and positive control antibody of the present application with the mouse TREM2 protein;
图1-9～图1-12为本申请全人抗体和阳性对照抗体与食蟹猴TREM2蛋白的结合检测结果图；Figures 1-9 to 1-12 are graphs showing the binding test results of the fully human antibody and the positive control antibody of the present application with the cynomolgus monkey TREM2 protein;
图2-1～图2-3为本申请全人抗体和阳性对照抗体与人TREM2过表达细胞株的结合检测结果图； Figures 2-1 to 2-3 are the results of binding detection of the fully human antibody and positive control antibody of the present application with human TREM2 overexpressing cell lines;
图2-4～图2-8为本申请全人抗体和阳性对照抗体与小鼠TREM2过表达细胞株的结合检测结果图；Figures 2-4 to 2-8 are graphs showing the binding test results of the fully human antibody and positive control antibody of the present application with the mouse TREM2 overexpressing cell line;
图2-9～图2-10为本申请全人抗体和阳性对照抗体与食蟹猴TREM2过表达细胞株的结合检测结果图；Figures 2-9 to 2-10 are the results of the binding test between the fully human antibody and the positive control antibody of the present application and the cynomolgus monkey TREM2 overexpressing cell line;
图3-1～图3-3为本申请全人抗体的ADCC效应结果图；Figures 3-1 to 3-3 are ADCC effect results of the fully human antibodies of the present application;
图4-1为本申请全人抗体的体内药效肿瘤生长曲线图；FIG4-1 is a graph showing the in vivo efficacy tumor growth curve of the fully human antibody of the present application;
图4-2为本申请全人抗体的体内药效体重变化趋势图。Figure 4-2 is a graph showing the in vivo efficacy and body weight change trends of the fully human antibodies of the present application.
为使本申请更加容易理解，下面将结合实施例来详细说明本申请，这些实施例仅起说明性作用，而并不会限制本申请的应用范围。In order to make the present application easier to understand, the present application will be described in detail below in conjunction with embodiments. These embodiments are only for illustration and do not limit the scope of application of the present application.
除非另有说明，本文使用的术语“包含、包括和含有(comprise、comprises和comprising)”或其等同物(contain、contains、containing、include、includes、including)为开放式表述，意味着除所列出的要素、组分和步骤外，还可涵盖其它未指明的要素、组分和步骤。Unless otherwise specified, the terms "comprise, comprises and comprising" or their equivalents (contain, contains, containing, include, includes, including) used herein are open-ended expressions and mean that in addition to the listed elements, components and steps, other unspecified elements, components and steps may also be included.
除非另有说明，本文所使用的表示成分的量、测量值或反应条件的所有数字应理解为在所有情况下均由术语“约”修饰。当与百分比相连时，术语“约”可以表示例如±1％、优选±0.5％、更优选±0.1％。Unless otherwise indicated, all numbers used herein expressing the amounts of ingredients, measurements or reaction conditions are to be understood as being modified in all cases by the term "about". When connected with a percentage, the term "about" can mean, for example, ±1%, preferably ±0.5%, more preferably ±0.1%.
除非上下文另有明确指示，本文中的单数术语涵盖复数的指示对象，反之亦然。类似地，除非上下文另有明确指示，本文中的词语“或”意在包括“和”。除非另有说明，本文使用的术语“任选的”或“任选地”表示其所修饰的对象或事件存在或不存在、或者发生或不发生。Unless the context clearly indicates otherwise, singular terms herein include plural referents, and vice versa. Similarly, the word "or" herein is intended to include "and" unless the context clearly indicates otherwise. Unless otherwise indicated, the term "optional" or "optionally" as used herein means that the object or event it modifies exists or does not exist, or occurs or does not occur.
除非另有说明，本文使用的术语“治疗”意味着在给药后可以抑制、遏制、减轻、改善、减缓、缓解或消除疾病或其相关症状，可以迟滞、延缓、减慢、中止或终止疾病或其相关症状的进展，或者可以阻止、控制或减少疾病或其相关症状的复发。Unless otherwise indicated, the term "treatment" as used herein means that after administration, the disease or its related symptoms can be inhibited, contained, alleviated, improved, mitigated, relieved or eliminated, the progression of the disease or its related symptoms can be delayed, postponed, slowed, stopped or terminated, or the recurrence of the disease or its related symptoms can be prevented, controlled or reduced.
除非另有说明，本文使用的术语“受试者”涵盖任何脊椎动物，例如哺乳动物和非哺乳动物，诸如人、非人灵长类动物、羊、犬、猫、马、牛、鸡、猪、鼠等。优选地，本申请中的受试者是人。本文使用的术语“受试者”可与术语“患者”互换使用。Unless otherwise indicated, the term "subject" as used herein encompasses any vertebrate, e.g., mammals and non-mammals, such as humans, non-human primates, sheep, dogs, cats, horses, cows, chickens, pigs, mice, etc. Preferably, the subject in the present application is a human. The term "subject" as used herein can be used interchangeably with the term "patient".
术语“互补决定区”可以指在抗体的可变区中赋予抗原结合特异性的部分，并且可以指在免疫球蛋白的重链或轻链的高可变区中发现的氨基酸序列。重链可以包括三个互补决定区CDR(HCDR1、HCDR2和HCDR3；以及轻链可以包括三个互补决定区CDR(LCDR1、LCDR2和LCDR3)。所述CDR可以提供在抗体与其抗原或抗原表位的结合中起重要作用的接触残基。The term "complementarity determining region" may refer to the portion of the variable region of an antibody that confers antigen binding specificity, and may refer to an amino acid sequence found in the hypervariable region of a heavy or light chain of an immunoglobulin. The heavy chain may include three complementary determining regions CDR (HCDR1, HCDR2, and HCDR3; and the light chain may include three complementary determining regions CDR (LCDR1, LCDR2, and LCDR3). The CDRs may provide contact residues that play an important role in the binding of an antibody to its antigen or antigenic epitope.
在本文中，“全人源抗体”或“全人抗体”可以互换使用。“全人源抗体”是指通过转基因等技术将编码抗体的人类基因转移至基因工程改造的抗体基因缺失动物中，由此得到的人类抗体。目前已建立多种方法生产全人源抗体，包括噬菌体展示技术、转基因小鼠技术、核糖体展示技术和RNA-多肽技术等。In this article, "fully human antibody" or "fully human antibody" can be used interchangeably. "Fully human antibody" refers to human antibodies obtained by transferring human genes encoding antibodies into genetically engineered antibody gene-deficient animals through transgenic and other technologies. Currently, a variety of methods have been established to produce fully human antibodies, including phage display technology, transgenic mouse technology, ribosome display technology, and RNA-peptide technology.
当提及抗体时，除非另有说明，其为分离的抗体，本文中的术语“分离的”表示该抗体基本上不含在天然状态下与其结合的其他细胞组分，例如，分离的抗体可以是从天然或自然环境中移出的抗体。When referring to an antibody, unless otherwise indicated, it is an isolated antibody. The term "isolated" herein means that the antibody is substantially free of other cellular components with which it is naturally associated, for example, an isolated antibody can be an antibody removed from its native or natural environment.
在本文中，序列之间的同一性百分比(同源性程度)可以通过例如使用万维网(例如www.ncbi.nlm.nih.gov)上通常用于此目的的免费可用的计算机程序(例如具有默认设置的BLASTp或BLASTn)对两个序列进行比较来确定。In the present context, the percent identity (degree of homology) between sequences can be determined by comparing two sequences, for example, using freely available computer programs commonly used for this purpose on the World Wide Web (eg www.ncbi.nlm.nih.gov), such as BLASTp or BLASTn with default settings.
本申请第一方面提供了一种抗TREM2抗体或其抗原结合片段，其包含：In a first aspect, the present application provides an anti-TREM2 antibody or an antigen-binding fragment thereof, comprising:
(a)包含下述互补决定区的重链可变区：(a) a heavy chain variable region comprising the following complementarity determining regions:
HCDR1(重链CDR1)，其包含SEQ ID NO:1、9、17、25、31、39、51、59或67所示的氨基酸序列，或与SEQ ID NO：1、9、17、25、31、39、51、59或67所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；HCDR1 (heavy chain CDR1) comprising the amino acid sequence of SEQ ID NO: 1, 9, 17, 25, 31, 39, 51, 59 or 67, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 1, 9, 17, 25, 31, 39, 51, 59 or 67;
HCDR2(重链CDR2)，其包含SEQ ID NO:2、10、18、26、32、52、60或68所示的氨基酸序列，或与SEQ ID NO：2、10、18、26、32、52、60或68所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；HCDR2 (heavy chain CDR2) comprising the amino acid sequence of SEQ ID NO: 2, 10, 18, 26, 32, 52, 60 or 68, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 2, 10, 18, 26, 32, 52, 60 or 68;
HCDR3(重链CDR3)，其包含SEQ ID NO:3、11、19、27、33、40、53、61或69所示的氨基酸序列，或与SEQ ID NO：3、11、19、27、33、40、53、61或69所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；和/或HCDR3 (heavy chain CDR3) comprising the amino acid sequence shown in SEQ ID NO: 3, 11, 19, 27, 33, 40, 53, 61 or 69, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 3, 11, 19, 27, 33, 40, 53, 61 or 69; and/or
(b)包含下述互补决定区的轻链可变区：(b) a light chain variable region comprising the following complementarity determining regions:
LCDR1(轻链CDR1)，其包含SEQ ID NO:4、12、20、34、41、46、54、62或73所示的氨基酸序列， 或与SEQ ID NO：4、12、20、34、41、46、54、62或73所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；LCDR1 (light chain CDR1) comprises the amino acid sequence shown in SEQ ID NO: 4, 12, 20, 34, 41, 46, 54, 62 or 73, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:4, 12, 20, 34, 41, 46, 54, 62 or 73;
LCDR2(轻链CDR2)，其包含SEQ ID NO:5、13、21、35、42、47、55或63所示的氨基酸序列，或与SEQ ID NO：5、13、21、35、42、47、55或63所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；LCDR2 (light chain CDR2) comprising the amino acid sequence shown in SEQ ID NO: 5, 13, 21, 35, 42, 47, 55 or 63, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 5, 13, 21, 35, 42, 47, 55 or 63;
LCDR3(轻链CDR3)，其包含SEQ ID NO:6、14、22、28、36、43、48、56、64、70或74所示的氨基酸序列，或与SEQ ID NO：6、14、22、28、36、43、48、56、64、70或74所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。LCDR3 (light chain CDR3), which comprises the amino acid sequence shown in SEQ ID NO: 6, 14, 22, 28, 36, 43, 48, 56, 64, 70 or 74, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 6, 14, 22, 28, 36, 43, 48, 56, 64, 70 or 74.
在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：重链可变区，所述重链可变区包含SEQ ID NOs：7、15、23、29、37、44、49、57、65、71或75所示的氨基酸序列或与SEQ ID NOs：7、15、23、29、37、44、49、57、65、71或75所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；和/或In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NOs: 7, 15, 23, 29, 37, 44, 49, 57, 65, 71 or 75, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence as shown in SEQ ID NOs: 7, 15, 23, 29, 37, 44, 49, 57, 65, 71 or 75; and/or
轻链可变区，所述轻链可变区包含SEQ ID NOs：8、16、24、30、38、45、50、58、66、72或76所示的氨基酸序列或与SEQ ID NOs：8、16、24、30、38、45、50、58、66、72或76所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。A light chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NOs: 8, 16, 24, 30, 38, 45, 50, 58, 66, 72 or 76, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence as shown in SEQ ID NOs: 8, 16, 24, 30, 38, 45, 50, 58, 66, 72 or 76.
在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：1、2和3所示的氨基酸序列，或与SEQ ID NOs：1、2和3所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：4、5和6所示的氨基酸序列，或与SEQ ID NOs：4、5和6所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(1) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 6, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 4, 5 and 6;
(2)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：9、10和11所示的氨基酸序列，或与SEQ ID NOs：9、10和11所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：12、13和14所示的氨基酸序列，或与SEQ ID NOs：12、13和14所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(2) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 9, 10 and 11, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% similarity to the amino acid sequences shown in SEQ ID NOs: 9, 10 and 11. wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 12, 13 and 14, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 12, 13 and 14;
(3)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：17、18和19所示的氨基酸序列，或与SEQ ID NOs：17、18和19所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：20、21和22所示的氨基酸序列，或与SEQ ID NOs：20、21和22所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(3) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 19, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 20, 21 and 22, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 20, 21 and 22;
(4)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：25、26和27所示的氨基酸序列，或与SEQ ID NOs：25、26和27所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：4、5和28所示的氨基酸序列，或与SEQ ID NOs：4、5和28所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(4) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96% similarity to the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27. , 97%, 98% or 99% identical amino acid sequences; said LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28;
(5)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：31、32和33所示的氨基酸序列，或与SEQ ID NOs：31、32和33所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs： 34、35和36所示的氨基酸序列，或与SEQ ID NOs：34、35和36所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(5) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 31, 32 and 33, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 31, 32 and 33, respectively; and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 34, 35 and 36, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NOs: 34, 35 and 36;
(6)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：39、18和40所示的氨基酸序列，或与SEQ ID NOs：39、18和40所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：41、42和43所示的氨基酸序列，或与SEQ ID NOs：41、42和43所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(6) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 39, 18 and 40, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 130%, 131%, 132%, 133%, 137%, 138%, 139%, wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 41, 42 and 43, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 41, 42 and 43;
(7)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：17、18和11所示的氨基酸序列，或与SEQ ID NOs：17、18和11所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：46、47和48所示的氨基酸序列，或与SEQ ID NOs：46、47和48所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(7) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 1112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 119, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 46, 47 and 48, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 46, 47 and 48;
(8)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：51、52和53所示的氨基酸序列，或与SEQ ID NOs：51、52和53所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：54、55和56所示的氨基酸序列，或与SEQ ID NOs：54、55和56所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(8) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 51, 52 and 53, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153 wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 54, 55 and 56, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 54, 55 and 56;
(9)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：59、60和61所示的氨基酸序列，或与SEQ ID NOs：59、60和61所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：62、63和64所示的氨基酸序列，或与SEQ ID NOs：62、63和64所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(9) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 59, 60 and 61, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, wherein the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 62, 63 and 64, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 62, 63 and 64;
(10)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：67、68和69所示的氨基酸序列，或与SEQ ID NOs：67、68和69所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：4、5和70所示的氨基酸序列，或与SEQ ID NOs：4、5和70所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；或(10) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 67, 68 and 69, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96% similarity to the amino acid sequences shown in SEQ ID NOs: 67, 68 and 69. , 97%, 98% or 99% identical amino acid sequences; said LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 70, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences shown in SEQ ID NOs: 4, 5 and 70; or
(11)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3；所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NOs：17、18和11所示的氨基酸序列，或与SEQ ID NOs：17、18和11所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NOs：73、47和74所示的氨基酸序列，或与SEQ ID NOs：73、47和74所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。(11) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively, or have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 1112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 119, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, The LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 73, 47 and 74, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences shown in SEQ ID NOs: 73, 47 and 74, respectively.
在本申请的一些具体实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some specific embodiments of the present application, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NO：1、2和3所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NO：4、5和6所示的氨基酸序列；或(1) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 6, respectively; or
(2)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NO：9、10和11所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NO：12、13和14所示的氨基酸序列；(2) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 9, 10 and 11, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 12, 13 and 14, respectively;
(3)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分 别具有SEQ ID NO：17、18和19所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NO：20、21和22所示的氨基酸序列；(3) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein HCDR1, HCDR2 and HCDR3 are The LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 20, 21 and 22, respectively;
(4)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NO：25、26和27所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NO：4、5和28所示的氨基酸序列；(4) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28, respectively;
(5)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NO：31、32和33所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NO：34、35和36所示的氨基酸序列；(5) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 31, 32 and 33, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 34, 35 and 36, respectively;
(6)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NO：39、18和40所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NO：41、42和43所示的氨基酸序列；(6) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 39, 18 and 40, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 41, 42 and 43, respectively;
(7)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NO：17、18和11所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NO：46、47和48所示的氨基酸序列；(7) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 46, 47 and 48, respectively;
(8)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NO：51、52和53所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NO：54、55和56所示的氨基酸序列；(8) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 51, 52 and 53, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 54, 55 and 56, respectively;
(9)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NO：59、60和61所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NO：62、63和64所示的氨基酸序列；(9) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 59, 60 and 61, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 62, 63 and 64, respectively;
(10)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NO：67、68和69所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NO：4、5和70所示的氨基酸序列；或(10) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 67, 68 and 69, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 4, 5 and 70, respectively; or
(11)HCDR1、HCDR2、HCDR3、LCDR1、LCDR2和LCDR3，所述HCDR1、HCDR2和HCDR3分别具有SEQ ID NO：17、18和11所示的氨基酸序列，所述LCDR1、LCDR2和LCDR3分别具有SEQ ID NO：73、47和74所示的氨基酸序列。(11) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 have the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively, and the LCDR1, LCDR2 and LCDR3 have the amino acid sequences shown in SEQ ID NOs: 73, 47 and 74, respectively.
在本申请的一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments of the present application, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)分别具有SEQ ID NOs：1、2和3所示的氨基酸序列的HCDR1、HCDR2和HCDR3；分别具有SEQ ID NOs：4、5和6所示的氨基酸序列的LCDR1、LCDR2和LCDR3；(1) HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 4, 5 and 6, respectively;
(2)分别具有SEQ ID NOs：17、18和19所示的氨基酸序列的HCDR1、HCDR2和HCDR3；分别具有SEQ ID NOs：20、21和22所示的氨基酸序列的LCDR1、LCDR2和LCDR3；或者(2) HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 17, 18 and 19, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 20, 21 and 22, respectively; or
(3)分别具有SEQ ID NOs：39、18和40所示的氨基酸序列的HCDR1、HCDR2和HCDR3；分别具有SEQ ID NOs：41、42和43所示的氨基酸序列的LCDR1、LCDR2和LCDR3。(3) HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 39, 18 and 40, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 41, 42 and 43, respectively.
在本申请的一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments of the present application, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)分别具有SEQ ID NOs：1、2和3所示的氨基酸序列的HCDR1、HCDR2和HCDR3；分别具有SEQ ID NOs：4、5和6所示的氨基酸序列的LCDR1、LCDR2和LCDR3；(1) HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 4, 5 and 6, respectively;
(2)分别具有SEQ ID NOs：17、18和19所示的氨基酸序列的HCDR1、HCDR2和HCDR3；分别具有SEQ ID NOs：20、21和22所示的氨基酸序列的LCDR1、LCDR2和LCDR3；(2) HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 17, 18 and 19, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 20, 21 and 22, respectively;
(3)分别具有SEQ ID NOs：25、26和27所示的氨基酸序列的HCDR1、HCDR2和HCDR3；分别具有SEQ ID NOs：4、5和28所示的氨基酸序列的LCDR1、LCDR2和LCDR3；或者(3) HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 25, 26 and 27, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 4, 5 and 28, respectively; or
(4)分别具有SEQ ID NOs：17、18和11所示的氨基酸序列的HCDR1、HCDR2和HCDR3；分别具有SEQ ID NOs：46、47和48所示的氨基酸序列的LCDR1、LCDR2和LCDR3。(4) HCDR1, HCDR2 and HCDR3 having the amino acid sequences shown in SEQ ID NOs: 17, 18 and 11, respectively; LCDR1, LCDR2 and LCDR3 having the amino acid sequences shown in SEQ ID NOs: 46, 47 and 48, respectively.
在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：7所示的氨基酸序列或与SEQ ID NO：7所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：8所示的氨基酸序列或与SEQ ID NO：8所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(1) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:7, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:7, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:8, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:8;
(2)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：15所示的氨基酸序列或与SEQ ID NO：15所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：16所示的氨基酸序列或与SEQ ID NO：16所示的氨基酸序列具有至少80％、81％、82％、83％、84％、 85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(2) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:15, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:15, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:16, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:16. amino acid sequences that are 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical;
(3)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：23所示的氨基酸序列或与SEQ ID NO：23所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：24所示的氨基酸序列或与SEQ ID NO：24所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(3) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:23, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:23, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:24, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:24;
(4)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：29所示的氨基酸序列或与SEQ ID NO：29所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：30所示的氨基酸序列或与SEQ ID NO：30所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(4) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:29, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:29, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:30, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:30;
(5)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：37所示的氨基酸序列或与SEQ ID NO：37所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：38所示的氨基酸序列或与SEQ ID NO：38所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(5) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:37, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:37, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:38, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:38;
(6)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：44所示的氨基酸序列或与SEQ ID NO：44所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：45所示的氨基酸序列或与SEQ ID NO：45所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(6) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:44, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:44, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:45, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:45;
(7)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：49所示的氨基酸序列或与SEQ ID NO：49所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：50所示的氨基酸序列或与SEQ ID NO：50所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(7) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:49, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:49, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:50, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:50;
(8)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：57所示的氨基酸序列或与SEQ ID NO：57所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：58所示的氨基酸序列或与SEQ ID NO：58所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(8) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:57, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:57, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:58, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:58;
(9)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：65所示的氨基酸序列或与SEQ ID NO：65所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：66所示的氨基酸序列或与SEQ ID NO：66所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(9) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:65, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:65, and the light chain variable region comprises the amino acid sequence of SEQ ID NO:66, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:66;
(10)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：71所示的氨基酸序列或与SEQ ID NO：71所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：72所示的氨基酸序列或与SEQ ID NO：72所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；或(10) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71 or is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 71. The light chain variable region comprises the amino acid sequence of SEQ ID NO: 72 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 72; or
(11)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：75所示的氨基酸序列或与SEQ ID NO：75所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：76所示的氨基酸序列或与SEQ ID NO：76所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。(11) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 75 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 75, and the light chain variable region comprises SEQ ID NO: NO:76 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:76.
在一些实施方式中，与SEQ ID NO：7、8、15、16、23、24、29、30、37、38、44、45、49、50、57、58、65、66、71、72、75或76所示的氨基酸序列相比，具有至少80％同一性的氨基酸序列中的差异的氨基酸主要或全部位于FR区中。In some embodiments, the different amino acids in an amino acid sequence with at least 80% identity compared to the amino acid sequence shown in SEQ ID NO: 7, 8, 15, 16, 23, 24, 29, 30, 37, 38, 44, 45, 49, 50, 57, 58, 65, 66, 71, 72, 75 or 76 are mainly or entirely located in the FR region.
在本申请的一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments of the present application, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：7所示的氨基酸序列或与SEQ ID NO：7所示的氨基酸序列具有至少80％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：8所示的氨基酸序列或与SEQ ID NO：8所示的氨基酸序列具有至少80％同一性的氨基酸序列；(1) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 7 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 7, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 8 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 8;
(2)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：23所示的氨基酸序列或与SEQ ID NO：23所示的氨基酸序列具有至少80％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：24所示的氨基酸序列或与SEQ ID NO：24所示的氨基酸序列具有至少80％同一性的氨基酸序列；或者(2) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 23, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 24 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 24; or
(3)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：44所示的氨基酸序列或与SEQ ID NO：44所示的氨基酸序列具有至少80％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：45所示的氨基酸序列或与SEQ ID NO：45所示的氨基酸序列具有至少80％同一性的氨基酸序列；(3) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 44 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 44, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 45 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 45;
其中，与SEQ ID NO：7、8、23、24、44或45所示的氨基酸序列相比，具有至少80％同一性的氨基酸序列中的差异的氨基酸全部位于FR区中。Among them, compared with the amino acid sequence shown in SEQ ID NO: 7, 8, 23, 24, 44 or 45, the different amino acids in the amino acid sequence with at least 80% identity are all located in the FR region.
在本申请的一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments of the present application, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：7所示的氨基酸序列或与SEQ ID NO：7所示的氨基酸序列具有至少80％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：8所示的氨基酸序列或与SEQ ID NO：8所示的氨基酸序列具有至少80％同一性的氨基酸序列；(1) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 7 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 7, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 8 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 8;
(2)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：23所示的氨基酸序列或与SEQ ID NO：23所示的氨基酸序列具有至少80％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：24所示的氨基酸序列或与SEQ ID NO：24所示的氨基酸序列具有至少80％同一性的氨基酸序列；(2) a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 23, and the light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 24 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 24;
(3)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：29所示的氨基酸序列或与SEQ ID NO：29所示的氨基酸序列具有至少80％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：30所示的氨基酸序列或与SEQ ID NO：30所示的氨基酸序列具有至少80％同一性的氨基酸序列；或者(3) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 29, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 30 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 30; or
(4)重链可变区和轻链可变区，所述重链可变区包含SEQ ID NO：49所示的氨基酸序列或与SEQ ID NO：49所示的氨基酸序列具有至少80％同一性的氨基酸序列，所述轻链可变区包含SEQ ID NO：50所示的氨基酸序列或与SEQ ID NO：50所示的氨基酸序列具有至少80％同一性的氨基酸序列；(4) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 49 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 49, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 50 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 50;
其中，与SEQ ID NO：7、8、23、24、29、30、49或50所示的氨基酸序列相比，具有至少80％同一性的氨基酸序列中的差异的氨基酸全部位于FR区中。Among them, compared with the amino acid sequence shown in SEQ ID NO: 7, 8, 23, 24, 29, 30, 49 or 50, the different amino acids in the amino acid sequence with at least 80% identity are all located in the FR region.
上述CDR的范围已根据AbM编号规则进行了定义，但是本领域技术人员可以理解的是，根据ImMunoGenTics(IMGT)编号规则、Kabat编号规则、Chothia编号规则和Contact编号规则(基于对可用复杂晶体结构的分析)等中的任一种或多种进行编号的CDR序列也落入了本申请的保护范围内。The scope of the above-mentioned CDR has been defined according to the AbM numbering rules, but those skilled in the art will appreciate that CDR sequences numbered according to any one or more of the ImMunoGenTics (IMGT) numbering rules, the Kabat numbering rules, the Chothia numbering rules and the Contact numbering rules (based on analysis of available complex crystal structures) also fall within the scope of protection of the present application.
在本申请中，还可对抗TREM2抗体或其抗原结合片段进行保守修饰。保守修饰是指不会明显影响或改变抗体结合特性的氨基酸修饰。例如，保守氨基酸取代可为采用同类(具有类似的化学性质或者功能)的另一种氨基酸进行取代。作为示例，可根据氨基酸的侧链性质将其分为：(1)非极性氨基酸：Ala(A)、Val(V)、Leu(L)、Ile(I)、Pro(P)、Phe(F)、Trp(W)、Met(M)；(2)不带电荷的极性氨基酸：Gly(G)、Ser(S)、Thr(T)、Cys(C)、Tyr(Y)、Asn(N)、Gln(Q)；(3)酸性氨基酸：Asp(D)、Glu(E)；(4)碱性氨基酸：Lys(K)、Arg(R)、His(H)。或者，可基于共同的侧链特性将氨基酸分为：(1)疏水氨基酸：Met、Ala、Val、Leu、Ile；(2)中性亲水氨基酸：Cys、Ser、Thr、Asn、Gln；(3)酸性氨基酸：Asp、Glu；(4)碱性氨基酸：His、Lys、Arg；(5)影响链取向的氨基酸：Gly、Pro；(6)芳香族氨基酸：Trp、Tyr、Phe。In the present application, the anti-TREM2 antibody or its antigen-binding fragment may also be conservatively modified. Conservative modification refers to amino acid modification that does not significantly affect or change the binding properties of the antibody. For example, a conservative amino acid substitution may be substituted with another amino acid of the same type (having similar chemical properties or functions). As an example, the amino acids can be divided into the following categories according to their side chain properties: (1) non-polar amino acids: Ala (A), Val (V), Leu (L), Ile (I), Pro (P), Phe (F), Trp (W), Met (M); (2) uncharged polar amino acids: Gly (G), Ser (S), Thr (T), Cys (C), Tyr (Y), Asn (N), Gln (Q); (3) acidic amino acids: Asp (D), Glu (E); (4) basic amino acids: Lys (K), Arg (R), His (H). Alternatively, amino acids can be divided based on common side chain properties: (1) hydrophobic amino acids: Met, Ala, Val, Leu, Ile; (2) neutral hydrophilic amino acids: Cys, Ser, Thr, Asn, Gln; (3) acidic amino acids: Asp, Glu; (4) basic amino acids: His, Lys, Arg; (5) amino acids that affect chain orientation: Gly, Pro; (6) aromatic amino acids: Trp, Tyr, Phe.
本申请所述的抗TREM2抗体或其抗原结合片段及它们的变体均能够特异性地结合至TREM2。The anti-TREM2 antibodies or antigen-binding fragments thereof and variants thereof described in the present application are all capable of specifically binding to TREM2.
本申请抗TREM2抗体或其抗原结合片段为分离的抗TREM2抗体或其抗原结合片段。The anti-TREM2 antibody or antigen-binding fragment thereof of the present application is an isolated anti-TREM2 antibody or antigen-binding fragment thereof.
在一些实施方式中，所述抗体为鼠源抗体、猴源抗体、兔源抗体、嵌合抗体、人源化抗体、全人源抗体；所述抗体优选为全人源抗体。 In some embodiments, the antibody is a mouse antibody, a monkey antibody, a rabbit antibody, a chimeric antibody, a humanized antibody, or a fully human antibody; preferably, the antibody is a fully human antibody.
在一些实施方式中，所述抗TREM2抗体或其抗原结合片段选自单克隆抗体、多克隆抗体、中和抗体、拮抗性抗体、无海藻糖基化抗体、单特异性抗体、多特异性抗体、纳米抗体、Fab片段、Fab'片段、F(ab')2片段、Fd片段、Fv片段、dAb片段、分离的CDR区和scFv中至少一种。In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof is selected from at least one of a monoclonal antibody, a polyclonal antibody, a neutralizing antibody, an antagonistic antibody, an afucosylated antibody, a monospecific antibody, a multispecific antibody, a nanobody, a Fab fragment, a Fab' fragment, a F(ab') 2 fragment, a Fd fragment, a Fv fragment, a dAb fragment, an isolated CDR region, and a scFv.
在一些实施方式中，所述抗TREM2抗体为单克隆抗体。In some embodiments, the anti-TREM2 antibody is a monoclonal antibody.
在一些实施方式中，所述抗TREM2抗体或其抗原结合片段进一步包含重链恒定区和/或轻链恒定区；In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof further comprises a heavy chain constant region and/or a light chain constant region;
优选地，所述重链恒定区为IgG、IgA、IgM、IgE或IgD的重链恒定区或其变体；所述重链恒定区优选为IgG的重链恒定区或其变体，例如IgG1、IgG2、IgG3或IgG4的恒定区或其变体；所述重链恒定区更优选为IgG1的重链恒定区或其变体；例如人IgG1的重链恒定区或其变体，其氨基酸序列如SEQ ID NO：90或92所示。Preferably, the heavy chain constant region is the heavy chain constant region of IgG, IgA, IgM, IgE or IgD or a variant thereof; the heavy chain constant region is preferably the heavy chain constant region of IgG or a variant thereof, for example, the constant region of IgG1, IgG2, IgG3 or IgG4 or a variant thereof; the heavy chain constant region is more preferably the heavy chain constant region of IgG1 or a variant thereof; for example, the heavy chain constant region of human IgG1 or a variant thereof, whose amino acid sequence is shown in SEQ ID NO: 90 or 92.
优选地，轻链恒定区为κ链或λ链的恒定区或其变体，优选人κ链的恒定区或其变体，其氨基酸序列如SEQ ID NO：91所示。Preferably, the light chain constant region is the constant region of a κ chain or a λ chain or a variant thereof, preferably the constant region of a human κ chain or a variant thereof, whose amino acid sequence is shown in SEQ ID NO: 91.
在一些实施方式中，所述重链恒定区包含全长重链恒定区或其片段，所述片段可选自CH1区、Fc区或CH3区。In some embodiments, the heavy chain constant region comprises a full-length heavy chain constant region or a fragment thereof, and the fragment can be selected from a CH1 region, an Fc region, or a CH3 region.
在一些实施方式中，所述重链恒定区或其变体包含具有一种或多种增强抗体依赖性细胞介导的细胞毒性(ADCC)的突变的IgG1 Fc区。In some embodiments, the heavy chain constant region or its variant comprises an IgG1 Fc region having one or more mutations that enhance antibody-dependent cell-mediated cytotoxicity (ADCC).
在一些实施方式中，所述重链恒定区或其变体包含IgG1 Fc区第234位、第235位和第243位突变，或第239、第330和第332位突变；在一些实施方式中，所述重链恒定区或其变体包含IgG1 Fc区第234位、第235位、第243位、第239、第330或第332位的至少一种突变。In some embodiments, the heavy chain constant region or its variant comprises mutations at positions 234, 235 and 243, or mutations at positions 239, 330 and 332 of the IgG1 Fc region; in some embodiments, the heavy chain constant region or its variant comprises at least one mutation at positions 234, 235, 243, 239, 330 or 332 of the IgG1 Fc region.
在一些具体实施方式中，IgG1 Fc区包含S239D、A330L和I332E的突变。在一些实施方式中，所述抗TREM2抗体或其抗原结合片段进一步包含具有S239D、A330L和I332E中的至少一种突变的IgG1 Fc区。In some embodiments, the IgG1 Fc region comprises mutations of S239D, A330L, and I332E. In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof further comprises an IgG1 Fc region having at least one mutation of S239D, A330L, and I332E.
在一些实施方案中，Fc区结合Fcγ受体，例如：FcγRI、FcγRIIa、FcγRIIb、FcγRIIc、FcγRIIIa或FcγRIIIb。In some embodiments, the Fc region binds an Fcγ receptor, e.g., FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, or FcγRIIIb.
在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含上述的重链可变区和轻链可变区、以及IgG1 Fc区。在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含上述的重链可变区和轻链可变区、以及IgG1 Fc区和人κ链的恒定区。在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含上述的重链可变区和轻链可变区、IgG1 Fc区、以及浓度比为2:1的人κ链的恒定区和人λ链的恒定区。在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含上述的重链可变区和轻链可变区、以及具有S239D、A330L和I332E中的至少一种突变的IgG1 Fc区。在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：(1)上述的重链可变区和轻链可变区，(2)具有S239D、A330L和I332E中的至少一种突变的IgG1 Fc区；以及(3)人κ链的恒定区。在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：(1)上述的重链可变区和轻链可变区，(2)具有S239D、A330L和I332E中的至少一种突变的IgG1 Fc区；以及(3)浓度比为2:1的人κ链的恒定区和人λ链的恒定区。In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises the heavy chain variable region and light chain variable region described above, and an IgG1 Fc region. In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises the heavy chain variable region and light chain variable region described above, and an IgG1 Fc region and a constant region of a human κ chain. In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises the heavy chain variable region and light chain variable region described above, an IgG1 Fc region, and a constant region of a human κ chain and a constant region of a human λ chain at a concentration ratio of 2:1. In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises the heavy chain variable region and light chain variable region described above, and an IgG1 Fc region having at least one mutation of S239D, A330L, and I332E. In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises: (1) the heavy chain variable region and light chain variable region described above, (2) an IgG1 Fc region having at least one mutation among S239D, A330L and I332E; and (3) a constant region of a human κ chain. In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises: (1) the heavy chain variable region and light chain variable region described above, (2) an IgG1 Fc region having at least one mutation among S239D, A330L and I332E; and (3) a constant region of a human κ chain and a constant region of a human λ chain at a concentration ratio of 2:1.
在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
重链，所述重链包含SEQ ID NO：77、79、80、82、83、85或87所示的氨基酸序列或与SEQ ID NO：77、79、80、82、83、85或87所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；和/或A heavy chain comprising the amino acid sequence shown in SEQ ID NO: 77, 79, 80, 82, 83, 85 or 87, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 77, 79, 80, 82, 83, 85 or 87; and/or
轻链，所述轻链包含SEQ ID NO：78、81、84或86所示的氨基酸序列或与SEQ ID NO：78、81、84或86所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。A light chain comprising the amino acid sequence shown in SEQ ID NO: 78, 81, 84 or 86, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 78, 81, 84 or 86.
在一些实施方式中，与SEQ ID NO：77、79、80、82、83、85或87所示的氨基酸序列相比，具有至少80％同一性的氨基酸序列中的差异的氨基酸主要或全部位于所述重链的FR区和/或恒定区中；和/或与SEQ ID NO：78、81、84或86所示的氨基酸序列相比，具有至少80％同一性的氨基酸序列中的差异的氨基酸主要或全部位于所述轻链的FR区和/或恒定区中。In some embodiments, compared with the amino acid sequence shown in SEQ ID NO: 77, 79, 80, 82, 83, 85 or 87, the different amino acids in the amino acid sequence with at least 80% identity are mainly or entirely located in the FR region and/or constant region of the heavy chain; and/or compared with the amino acid sequence shown in SEQ ID NO: 78, 81, 84 or 86, the different amino acids in the amino acid sequence with at least 80% identity are mainly or entirely located in the FR region and/or constant region of the light chain.
在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)重链和轻链，所述重链包含SEQ ID NO：77所示的氨基酸序列或与SEQ ID NO：77所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：78所示的氨基酸序列或与SEQ ID NO：78所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列； (1) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:77, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:77; and the light chain comprises the amino acid sequence of SEQ ID NO:78, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:78;
(2)重链和轻链，所述重链包含SEQ ID NO：79所示的氨基酸序列或与SEQ ID NO：79所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：78所示的氨基酸序列或与SEQ ID NO：78所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(2) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:79, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:79; and the light chain comprises the amino acid sequence of SEQ ID NO:78, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:78;
(3)重链和轻链，所述重链包含SEQ ID NO：80所示的氨基酸序列或与SEQ ID NO：80所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：81所示的氨基酸序列或与SEQ ID NO：81所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(3) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:80, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:80; and the light chain comprises the amino acid sequence of SEQ ID NO:81, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:81;
(4)重链和轻链，所述重链包含SEQ ID NO：82所示的氨基酸序列或与SEQ ID NO：82所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：81所示的氨基酸序列或与SEQ ID NO：81所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(4) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:82, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:82; and the light chain comprises the amino acid sequence of SEQ ID NO:81, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:81;
(5)重链和轻链，所述重链包含SEQ ID NO：83所示的氨基酸序列或与SEQ ID NO：83所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：84所示的氨基酸序列或与SEQ ID NO：84所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；(5) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:83, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:83; and the light chain comprises the amino acid sequence of SEQ ID NO:84, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:84;
(6)重链和轻链，所述重链包含SEQ ID NO：85所示的氨基酸序列或与SEQ ID NO：85所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：86所示的氨基酸序列或与SEQ ID NO：86所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；或者(6) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:85 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:85; and the light chain comprises the amino acid sequence of SEQ ID NO:86 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:86; or
(7)重链和轻链，所述重链包含SEQ ID NO：87所示的氨基酸序列或与SEQ ID NO：87所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列；所述轻链包含SEQ ID NO：86所示的氨基酸序列或与SEQ ID NO：86所示的氨基酸序列具有至少80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列。(7) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 87 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 87; and the light chain comprises the amino acid sequence of SEQ ID NO: 86 or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 86.
在一些实施方式中，所述抗TREM2抗体或其抗原结合片段包含：In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof comprises:
(1)重链和轻链，所述重链包含SEQ ID NO：87所示的氨基酸序列或与SEQ ID NO：87所示的氨基酸序列具有至少80％同一性的氨基酸序列；所述轻链包含SEQ ID NO：86所示的氨基酸序列或与SEQ ID NO：86所示的氨基酸序列具有至少80％同一性的氨基酸序列；(1) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 87 or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 87; and the light chain comprises the amino acid sequence of SEQ ID NO: 86 or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 86;
(2)重链和轻链，所述重链包含SEQ ID NO：77所示的氨基酸序列或与SEQ ID NO：77所示的氨基酸序列具有至少80％同一性的氨基酸序列；所述轻链包含SEQ ID NO：78所示的氨基酸序列或与SEQ ID NO：78所示的氨基酸序列具有至少80％同一性的氨基酸序列；(2) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 77 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 77; and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 78 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 78;
(3)重链和轻链，所述重链包含SEQ ID NO：83所示的氨基酸序列或与SEQ ID NO：83所示的氨基酸序列具有至少80％同一性的氨基酸序列；所述轻链包含SEQ ID NO：84所示的氨基酸序列或与SEQ ID NO：84所示的氨基酸序列具有至少80％同一性的氨基酸序列；或者(3) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 83 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 83; and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 84 or an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 84; or
(4)重链和轻链，所述重链包含SEQ ID NO：82所示的氨基酸序列或与SEQ ID NO：82所示的氨基酸序列具有至少80％同一性的氨基酸序列；所述轻链包含SEQ ID NO：81所示的氨基酸序列或与SEQ ID NO：81所示的氨基酸序列具有至少80％同一性的氨基酸序列。(4) a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 82 or an amino acid sequence that is at least 80% identical to the amino acid sequence shown in SEQ ID NO: 82; and the light chain comprises the amino acid sequence shown in SEQ ID NO: 81 or an amino acid sequence that is at least 80% identical to the amino acid sequence shown in SEQ ID NO: 81.
在一些实施方式中，所述抗体为多特异性抗体，所述多特异性抗体特异性结合TREM2，并且额外地特异性结合一个或多个其他靶标。In some embodiments, the antibody is a multispecific antibody that specifically binds TREM2 and additionally specifically binds one or more other targets.
优选地，所述多特异性抗体还包含至少一种具有针对第二靶标的第二结合特异性的第二抗体。优选地，所述多特异性抗体为双特异性抗体、三特异性抗体或四特异性抗体。更优选地，所述多特异性抗体为双特异性抗体。Preferably, the multispecific antibody further comprises at least one second antibody having a second binding specificity for a second target. Preferably, the multispecific antibody is a bispecific antibody, a trispecific antibody or a tetraspecific antibody. More preferably, the multispecific antibody is a bispecific antibody.
作为非限制性示例，所述抗TREM2抗体或其抗原结合片段具有以下一个或多个特性：As non-limiting examples, the anti-TREM2 antibody or antigen-binding fragment thereof has one or more of the following properties:
(1)能够结合至人TREM2；(1) Ability to bind to human TREM2;
(2)能够结合至鼠TREM2； (2) able to bind to mouse TREM2;
(3)能够结合至食蟹猴TREM2；(3) able to bind to cynomolgus monkey TREM2;
(4)与一种或多种TREM2配体竞争结合TREM2(4) Compete with one or more TREM2 ligands for binding to TREM2
(5)抑制和/或阻断TREM2与配体的结合；(5) inhibiting and/or blocking the binding of TREM2 to ligands;
(6)抑制和/或阻断由TREM2结合配体介导的细胞内信号传导；(6) inhibiting and/or blocking intracellular signal transduction mediated by TREM2 binding ligand;
(7)诱导细胞因子或趋化因子分泌；(7) Induce cytokine or chemokine secretion;
(8)具有抗体依赖性细胞介导的细胞毒性(ADCC)活性、补体依赖性细胞毒性(CDC)活性和抗体介导的吞噬作用(ADCP)活性中的至少一种；(8) having at least one of antibody-dependent cell-mediated cytotoxicity (ADCC) activity, complement-dependent cytotoxicity (CDC) activity, and antibody-mediated phagocytosis (ADCP) activity;
(9)增强免疫应答；(9) Enhance immune response;
(10)增强的ADCC活性。(10) Enhanced ADCC activity.
在一些实施方案中，所述一种或多种TREM2配体选自大肠杆菌细胞、凋亡细胞、核酸、阴离子性脂质、两性离子性脂质、带负电的磷脂、磷脂酰丝氨酸、硫苷脂、磷脂酰胆碱(phosphatidylcholin)、鞘磷脂、膜磷脂、脂化蛋白、蛋白脂质、脂化肽及脂化β淀粉样肽。In some embodiments, the one or more TREM2 ligands are selected from Escherichia coli cells, apoptotic cells, nucleic acids, anionic lipids, zwitterionic lipids, negatively charged phospholipids, phosphatidylserine, sulfatides, phosphatidylcholin, sphingomyelin, membrane phospholipids, lipidated proteins, proteolipids, lipidated peptides, and lipidated amyloid β peptide.
在一些实施方案中，所述细胞因子或趋化因子选自IFN-γ、TNF-α、CXCL1或CXCL10。In some embodiments, the cytokine or chemokine is selected from IFN-γ, TNF-α, CXCL1, or CXCL10.
在一些实施方案中，所述抗TREM2抗体或其抗原结合片段能够对TREM2+髓系细胞进行特异性杀灭、消减或使其丧失能力。任选地，所述TREM2+髓系细胞为非刺激性髓系细胞，例如：CD45+、HLA-DR+、CD11c+、CD14+和BDCA3-非刺激性髓系细胞；任选地，所述TREM2+髓系细胞为肿瘤内髓系细胞。任选地，所述TREM2+细胞选自树突细胞、巨噬细胞、单核细胞和嗜中性白细胞；所述巨噬细胞优选为肿瘤相关巨噬细胞(TAM)。In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof is capable of specifically killing, depleting or disabling TREM2+ myeloid cells. Optionally, the TREM2+ myeloid cells are non-stimulatory myeloid cells, for example: CD45+, HLA-DR+, CD11c+, CD14+ and BDCA3- non-stimulatory myeloid cells; Optionally, the TREM2+ myeloid cells are intratumoral myeloid cells. Optionally, the TREM2+ cells are selected from dendritic cells, macrophages, monocytes and neutrophils; the macrophages are preferably tumor-associated macrophages (TAMs).
在一些具体实施方案中，所述抗TREM2抗体或其抗原结合片段具有ADCC活性。ADCC可在抗体结合病原性或致肿瘤性靶标细胞的表面上的抗原时发生。In some embodiments, the anti-TREM2 antibody or antigen-binding fragment thereof has ADCC activity. ADCC can occur when the antibody binds to an antigen on the surface of a pathogenic or tumorigenic target cell.
为评估目标分子的ADCC效应的活性，可进行诸如美国专利号5,500,362或5,821,337中所述的体外ADCC测定。可用于所述测定的效应细胞包括外周血液单个核细胞(PBMC)和天然杀伤(NK)细胞。或者或另外，可在体内，例如在诸如Clynes等,Proc.Natl.Acad.Sci.(USA)95:652-656(1998)中公开的动物模型中评估目标分子的ADCC效应的活性。To evaluate the activity of the ADCC effect of the target molecule, an in vitro ADCC assay such as described in U.S. Pat. No. 5,500,362 or 5,821,337 can be performed. Effector cells that can be used for the assay include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells. Alternatively or additionally, the activity of the ADCC effect of the target molecule can be evaluated in vivo, for example, in an animal model such as disclosed in Clynes et al., Proc. Natl. Acad. Sci. (USA) 95: 652-656 (1998).
本申请第二方面提供了一种核酸分子，所述核酸分子编码本申请第一方面所述的抗TREM2抗体或其抗原结合片段。The second aspect of the present application provides a nucleic acid molecule, which encodes the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application.
在一些实施方式中，所述核酸分子包括分别编码本申请所述的抗TREM2抗体或其抗原结合片段的重链可变区或/和轻链可变区的核酸分子。In some embodiments, the nucleic acid molecule includes a nucleic acid molecule encoding the heavy chain variable region and/or the light chain variable region of the anti-TREM2 antibody or antigen-binding fragment thereof described in the present application, respectively.
本申请第三方面提供了重组载体，其包含本申请第二方面所述的核酸分子。The third aspect of the present application provides a recombinant vector, which comprises the nucleic acid molecule described in the second aspect of the present application.
术语“载体”是指用于在宿主细胞中表达靶基因的工具，例如质粒载体、粘粒载体和病毒载体如噬菌体载体、慢病毒载体、腺病毒载体、逆转录病毒载体和腺相关病毒载体。重组载体可以由以下构建或通过操纵以下来构建：质粒(例如，pSC101、pGV1106、pACYC177、ColE1、pKT230、pME290、pBR322、pUC8/9、pUC6、pBD9、pHC79、pIJ61、pLAFR1、pHV14、pGEX系列、pET系列、pUC19等)、噬菌体(例如，λgt4λB、λ8Charon、λΔz1、M13等)或病毒载体(例如，SV40等)，这是本领域中常用的。The term "vector" refers to a tool for expressing a target gene in a host cell, such as a plasmid vector, a cosmid vector, and a viral vector such as a phage vector, a lentiviral vector, an adenoviral vector, a retroviral vector, and an adeno-associated viral vector. The recombinant vector can be constructed by or by manipulating a plasmid (e.g., pSC101, pGV1106, pACYC177, ColE1, pKT230, pME290, pBR322, pUC8/9, pUC6, pBD9, pHC79, pIJ61, pLAFR1, pHV14, pGEX series, pET series, pUC19, etc.), a phage (e.g., λgt4λB, λ 8 Charon, λΔz1, M13, etc.) or a viral vector (e.g., SV40, etc.), which is commonly used in the art.
在重组载体中，核酸分子可以与启动子可操作地连接。术语“可操作地连接”旨在涉及目标核苷酸序列与表达调控序列(例如，启动子序列)之间的功能连接。当“可操作地连接”时，调控元件可以控制目标多核苷酸的转录和/或翻译。In a recombinant vector, a nucleic acid molecule can be operably linked to a promoter. The term "operably linked" is intended to refer to a functional connection between a target nucleotide sequence and an expression control sequence (e.g., a promoter sequence). When "operably linked", a regulatory element can control transcription and/or translation of a target polynucleotide.
在一些实施方式中，所述载体为克隆载体；在另一些实施方式中，所述载体为表达载体。所述表达载体可任选地为能够表达本申请所述抗TREM2抗体或其抗原结合片段、或其部分(moiety)的任意表达载体。In some embodiments, the vector is a cloning vector; in other embodiments, the vector is an expression vector. The expression vector may optionally be any expression vector capable of expressing the anti-TREM2 antibody or antigen-binding fragment thereof, or a moiety thereof described herein.
为了在宿主例如原核或真核细胞中使用，可以相应地构建重组载体。例如，当构建载体作为用于原核宿主的表达载体时，所述载体通常包括用于转录的强启动子(例如，pLλ启动子、CMV启动子、trp启动子、lac启动子、tac启动子、T7启动子等)、用于启动翻译的核糖体结合位点和转录/翻译终止序列。另一方面，用于真核宿主的表达载体包括可在真核细胞中操作的复制起点，例如f1复制起点、SV40复制起点、pMB1复制起点、腺病毒复制起点、AAV复制起点和BBV复制起点，但不限于此。另外，表达载体通常包括源自哺乳动物细胞基因组的启动子(例如金属硫蛋白启动子)或源自哺乳动物病毒的启动子(例如腺病毒晚期启动子、牛痘病毒7.5K启动子、SV40启动子、巨细胞病毒启动子、HSV的tk启动子等)，以及作为转录终止序列的多聚腺苷酸化序列。可以通过将重组载体引入合适的宿主细胞来制备重组细胞。本领域已知的任何宿主细胞都可以用于本公开中，只要它允许重组载体以稳定的方式连续克隆和表达。可用于本公开的原核宿主细胞的实例可以选自大肠杆菌(E.coli)如大肠杆菌JM109、大肠杆菌BL21、大肠杆菌RR1、大肠杆菌LE392、大肠杆菌B、大肠杆菌X 1776、大肠杆菌W3110，芽孢杆菌属菌种如枯草芽孢杆菌(Bacillus subtilis)和苏云金芽孢杆菌(Bacillus thuringiensis)，以及肠杆菌科菌株，例如鼠伤寒沙门氏菌(Salmonella typhimurium)、粘质沙雷氏菌(Serratia marcescens)和各种假单胞菌菌种。可用于转化的真核宿主细胞可以选自但不限于酿酒酵母(Saccharomyces cerevisiae)、昆虫细胞和动物细胞，例如Sp2/0、CHO(中国仓鼠卵巢)K1、CHO DG44、CHO S、CHO DXB11、CHO GS8KO、PER.C6、W138、BHK、COS87、293、HepG2、Huh7、3T3、RIN、MDCK等。可以使用相关领域中众所周知的方法将核酸分子或携带核酸分子的重组载体引入(转染)到宿主细胞中。例如，当宿主细胞是原核细胞时，可以使用CaCl2或电穿孔方法进行这种转染。对于真核宿主细胞，可以使用但不限于显微注射、磷酸钙沉淀、电穿孔、脂质体介导的转染或粒子轰击来实现基因引入。In order to use in a host such as a prokaryotic or eukaryotic cell, a recombinant vector can be constructed accordingly. For example, when constructing a vector as an expression vector for a prokaryotic host, the vector generally includes a strong promoter for transcription (e.g., pLλ promoter, CMV promoter, trp promoter, lac promoter, tac promoter, T7 promoter, etc.), a ribosome binding site for initiating translation, and a transcription/translation termination sequence. On the other hand, an expression vector for a eukaryotic host includes a replication origin that can be operated in a eukaryotic cell, such as f1 replication origin, SV40 replication origin, pMB1 replication origin, adenovirus replication origin, AAV replication origin, and BBV replication origin, but is not limited thereto. In addition, the expression vector generally includes a promoter derived from a mammalian cell genome (e.g., a metallothionein promoter) or a promoter derived from a mammalian virus (e.g., adenovirus late promoter, vaccinia virus 7.5K promoter, SV40 promoter, cytomegalovirus promoter, tk promoter of HSV, etc.), and a polyadenylation sequence as a transcription termination sequence. Recombinant cells can be prepared by introducing a recombinant vector into a suitable host cell. Any host cell known in the art can be used in the present disclosure as long as it allows for continuous cloning and expression of the recombinant vector in a stable manner. Examples of prokaryotic host cells that can be used in the present disclosure can be selected from Escherichia coli (E. coli) such as E. coli JM109, E. coli BL21, E. coli RR1, E. coli LE392, E. coli B, E. coli X1776, E. coli W3110, Bacillus species such as Bacillus subtilis (Bacillus subtilis) and Bacillus thuringiensis (Bacillus thuringiensis), and Enterobacteriaceae strains, such as Salmonella typhimurium (Salmonella typhimurium), Serratia marcescens (Serratia marcescens) and various Pseudomonas species. The eukaryotic host cell that can be used for transformation can be selected from but not limited to Saccharomyces cerevisiae (Saccharomyces cerevisiae), insect cells and animal cells, such as Sp2/0, CHO (Chinese hamster ovary) K1, CHO DG44, CHO S, CHO DXB11, CHO GS 8 KO, PER.C6, W138, BHK, COS 8 7, 293, HepG2, Huh7, 3T3, RIN, MDCK etc. Nucleic acid molecules or recombinant vectors carrying nucleic acid molecules can be introduced (transfected) into host cells using methods well known in the relevant art. For example, when the host cell is a prokaryotic cell, CaCl2 or electroporation methods can be used to carry out this transfection. For eukaryotic host cells, gene introduction can be achieved using, but not limited to, microinjection, calcium phosphate precipitation, electroporation, liposome-mediated transfection, or particle bombardment.
为了选择转化的宿主细胞，可以根据本领域中众所周知的方法利用与选择标志物相关的表型。例如，当所述选择标志物是赋予某些抗生素抗性的基因时，宿主细胞可以在存在抗生素的情况下在培养基中生长以选择目标转化体。In order to select transformed host cells, the phenotype associated with the selection marker can be utilized according to methods well known in the art. For example, when the selection marker is a gene that confers resistance to certain antibiotics, host cells can be grown in culture medium in the presence of antibiotics to select target transformants.
本申请第四方面提供了宿主细胞，其包含本申请第二方面所述的核酸分子或本申请第三方面所述的重组载体。The fourth aspect of the present application provides a host cell, which comprises the nucleic acid molecule described in the second aspect of the present application or the recombinant vector described in the third aspect of the present application.
本申请中的宿主细胞可为用于表达抗TREM2抗体或其抗原结合片段、或其部分(moiety)的任何适当的宿主细胞。在一些实施方式中，所述宿主细胞为原核细胞。在另一些实施方式中，所述宿主细胞为真核细胞。在一些实施方式中，所述宿主细胞选自酵母细胞、哺乳动物细胞或适用于制备抗体或其抗原结合片段、或其部分(moiety)的其他细胞。哺乳动物细胞例如为中国仓鼠卵巢细胞(CHO)、人肾上皮细胞系293细胞或293T细胞。在一些实施方式中，所述宿主细胞为重组CHO细胞。The host cell in the present application may be any suitable host cell for expressing an anti-TREM2 antibody or its antigen-binding fragment, or a portion thereof (moiety). In some embodiments, the host cell is a prokaryotic cell. In other embodiments, the host cell is a eukaryotic cell. In some embodiments, the host cell is selected from yeast cells, mammalian cells, or other cells suitable for preparing antibodies or their antigen-binding fragments, or a portion thereof (moiety). Mammalian cells are, for example, Chinese hamster ovary cells (CHO), human kidney epithelial cell line 293 cells or 293T cells. In some embodiments, the host cell is a recombinant CHO cell.
本申请第五方面提供了制备抗TREM2抗体或其抗原结合片段的方法，其包括培养本申请第四方面所述的宿主细胞。在一些实施方式中，所述方法还包括对所述宿主细胞的表达产物进行分离和纯化，例如采用Protein A亲和柱纯化。The fifth aspect of the present application provides a method for preparing an anti-TREM2 antibody or an antigen-binding fragment thereof, which comprises culturing the host cell described in the fourth aspect of the present application. In some embodiments, the method further comprises isolating and purifying the expression product of the host cell, for example, using a Protein A affinity column for purification.
本申请第一方面所述的抗TREM2抗体或其抗原结合片段、本申请第二方面所述的核酸分子、本申请第三方面所述的重组载体或本申请第四方面所述的宿主细胞在制备药物中的用途。The use of the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application, the nucleic acid molecule described in the second aspect of the present application, the recombinant vector described in the third aspect of the present application, or the host cell described in the fourth aspect of the present application in the preparation of a drug.
在一些实施方式中，所述药物为预防和/或治疗TREM2相关的疾病或病症的药物。在一些实施方式中，所述TREM2相关的疾病或病症为癌症。在一些实施方式中，所述TREM2相关的疾病或病症为脂质相关疾病或病症。在一些实施方式中，所述TREM2相关的疾病或病症为纤维化疾病或病症。在一些实施方式中，所述药物为调节受试者的免疫应答的药物。在一些具体实施方式中，所述药物为增加受试者的抗肿瘤免疫应答的药物和/或诱导受试者的记忆免疫应答的药物。In some embodiments, the drug is a drug for preventing and/or treating a disease or condition associated with TREM2. In some embodiments, the disease or condition associated with TREM2 is cancer. In some embodiments, the disease or condition associated with TREM2 is a lipid-related disease or condition. In some embodiments, the disease or condition associated with TREM2 is a fibrotic disease or condition. In some embodiments, the drug is a drug for regulating the immune response of a subject. In some specific embodiments, the drug is a drug for increasing the anti-tumor immune response of a subject and/or a drug for inducing a memory immune response of a subject.
本申请第六方面提供了第一方面所述的抗TREM2抗体或其抗原结合片段、本申请第二方面所述的核酸分子、本申请第三方面所述的重组载体或本申请第四方面所述的宿主细胞胞在制备诊断TREM2相关的疾病或病症的试剂中的用途。The sixth aspect of the present application provides the use of the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect, the nucleic acid molecule described in the second aspect of the present application, the recombinant vector described in the third aspect of the present application, or the host cell described in the fourth aspect of the present application in the preparation of a reagent for diagnosing TREM2-related diseases or conditions.
在一些实施方式中，所述TREM2相关的疾病或病症为癌症。在一些实施方式中，所述TREM2相关的疾病或病症为脂质相关疾病或病症。在一些实施方式中，所述TREM2相关的疾病或病症为纤维化疾病或病症。在一些实施方式中，所述试剂能够调节受试者的免疫应答。在一些具体实施方式中，所述试剂能够增加受试者的抗肿瘤免疫应答和/或诱导受试者的记忆免疫应答。In some embodiments, the TREM2-related disease or condition is cancer. In some embodiments, the TREM2-related disease or condition is a lipid-related disease or condition. In some embodiments, the TREM2-related disease or condition is a fibrotic disease or condition. In some embodiments, the agent is capable of modulating an immune response in a subject. In some specific embodiments, the agent is capable of increasing an anti-tumor immune response in a subject and/or inducing a memory immune response in a subject.
本申请第七方面提供了一种用于检测TREM2的试剂盒，其包含本申请第一方面所述的抗TREM2抗体或其抗原结合片段或本申请第二方面所述的核酸分子。The seventh aspect of the present application provides a kit for detecting TREM2, which comprises the anti-TREM2 antibody or its antigen-binding fragment described in the first aspect of the present application or the nucleic acid molecule described in the second aspect of the present application.
本申请第八方面提供了一种治疗或预防有需要的受试者的疾病或病症的方法，其包括向所述受试者施用本申请第一方面所述的抗TREM2抗体或其抗原结合片段。The eighth aspect of the present application provides a method for treating or preventing a disease or condition in a subject in need thereof, comprising administering the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application to the subject.
在一些实施方式中，所述疾病或病症为TREM2相关的疾病或病症。在一些实施方式中，所述TREM2相关的疾病或病症为癌症。在一些实施方式中，所述TREM2相关的疾病或病症为脂质相关疾病或病症。在一些实施方式中，所述TREM2相关的疾病或病症为纤维化疾病或病症。In some embodiments, the disease or condition is a TREM2-related disease or condition. In some embodiments, the TREM2-related disease or condition is cancer. In some embodiments, the TREM2-related disease or condition is a lipid-related disease or condition. In some embodiments, the TREM2-related disease or condition is a fibrotic disease or condition.
本申请第九方面提供了一种增强受试者的免疫应答的方法，其包括向所述受试者施用本申请第一方面所述的抗TREM2抗体或其抗原结合片段。The ninth aspect of the present application provides a method for enhancing the immune response of a subject, comprising administering the anti-TREM2 antibody or antigen-binding fragment thereof described in the first aspect of the present application to the subject.
优选地，所述增强受试者的免疫应答为增加受试者的抗肿瘤免疫应答和/或诱导受试者的记忆免疫应答。在一些具体的实施方式中，所述受试者为哺乳动物，例如人。在一些实施方式中，所述免疫应答是适应性免疫应答。在一些实施方式中，所述免疫应答是先天性免疫应答。Preferably, the enhancing the immune response of the subject is to increase the anti-tumor immune response of the subject and/or induce the memory immune response of the subject. In some specific embodiments, the subject is a mammal, such as a human. In some embodiments, the immune response is an adaptive immune response. In some embodiments, the immune response is an innate immune response.
本申请抗TREM2抗体或其抗原结合片段可杀伤的癌细胞包括但不限于来自以下的癌细胞：膀胱、血液、骨、骨髓、脑、乳腺、结肠、食管、胃肠、牙龈、头、肾、肝、肺、鼻咽、颈、卵巢、前列腺、皮肤、胃、胰腺、睾丸、舌头、宫颈或子宫。另外，癌症可具体地具有以下组织学类型，尽管不限于这些：肿瘤， 恶性肿瘤病。在一些实施方式中，所述癌症选自黑素瘤、肾癌、肝胆癌、头颈部鳞状癌(HNSC)、胰腺癌、结直肠癌、膀胱癌、胶质母细胞瘤、前列腺癌、肺癌、乳腺癌、卵巢癌、胃癌、食道癌、肾脏癌、白血病、淋巴瘤或间皮瘤。在一些具体实施方式中，所述癌症为结直肠癌。在一些具体实施方式中，所述癌症为乳腺癌。Cancer cells that can be killed by the anti-TREM2 antibodies or antigen-binding fragments thereof of the present application include, but are not limited to, cancer cells from the following: bladder, blood, bone, bone marrow, brain, breast, colon, esophagus, gastrointestinal tract, gums, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate, skin, stomach, pancreas, testicles, tongue, cervix or uterus. In addition, cancer may specifically have the following histological types, although not limited to these: tumor, Malignant tumor disease. In some embodiments, the cancer is selected from melanoma, renal cancer, hepatobiliary cancer, head and neck squamous cell carcinoma (HNSC), pancreatic cancer, colorectal cancer, bladder cancer, glioblastoma, prostate cancer, lung cancer, breast cancer, ovarian cancer, gastric cancer, esophageal cancer, kidney cancer, leukemia, lymphoma or mesothelioma. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is breast cancer.
在一些实施方式中，所述脂质相关疾病或病症选自肥胖症、脂肪肝病、心脏病、中风、动脉粥样硬化、糖尿病、骨关节炎、痛风、睡眠呼吸暂停或高血压。In some embodiments, the lipid-related disease or disorder is selected from obesity, fatty liver disease, heart disease, stroke, atherosclerosis, diabetes, osteoarthritis, gout, sleep apnea, or hypertension.
在一些实施方式中，所述纤维化疾病或病症是肝病。任选地，所述肝病为非酒精性脂肪性肝病(NAFLD)或非酒精性脂肪性肝炎(NASH)。In some embodiments, the fibrotic disease or disorder is a liver disease. Optionally, the liver disease is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).
本申请所述的抗TREM2抗体或其抗原结合片段的合适剂量可由临床医师根据如下因素确定，例如受试者的年龄、体重、性别、一般状况、疾病严重程度；它们可通过本领域已知的任何常规途径给予，例如胃肠外、静脉内、肌内、动脉内、鞘内、囊内、眶内、心内、皮内、腹膜内、经气管、皮下、表皮下、关节内、囊下、蛛网膜下、脊柱内、硬膜外和胸骨内施用。The appropriate dose of the anti-TREM2 antibodies or antigen-binding fragments thereof described herein can be determined by a clinician based on factors such as the age, weight, sex, general condition, and severity of the disease of the subject; they can be administered by any conventional route known in the art, such as parenteral, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal administration.
本领域技术人员可以根据本领域的普通知识对给药方案进行调整以提供最佳的所需应答(例如，临床应答)。例如，可以施用单次大剂量、可以随时间推移施用多个分剂量、或者可以随治疗情况的危急程度成比例降低或提高单次的剂量。例如，当将抗体作为缓释剂给药时，能够以降低的频率给药。Those skilled in the art can adjust the dosage regimen to provide the best desired response (e.g., clinical response) based on common knowledge in the art. For example, a single large dose can be administered, multiple divided doses can be administered over time, or the single dose can be reduced or increased in proportion to the severity of the treatment situation. For example, when the antibody is administered as a sustained release formulation, it can be administered at a reduced frequency.
实施例Example
下面将结合实施例对本申请的方案进行进一步的说明，但是本领域技术人员将能够理解的是，本申请的保护范围并不仅限于这些实施例。The scheme of the present application will be further described below in conjunction with embodiments, but those skilled in the art will appreciate that the protection scope of the present application is not limited to these embodiments.
除非另有特别说明，下述实施例中涉及的试剂、材料或仪器为可商购得到的。Unless otherwise specified, the reagents, materials or instruments involved in the following examples are commercially available.
实施例1、阳性对照抗体的表达Example 1. Expression of positive control antibody
以Pionyr Immunotherapeutics公司的TREM2抗体PY314作为阳性对照抗体，PY314的可变区氨基酸序列记载于WO2019118513，采用IgG1亚型，并通过重链恒定区S239D/I332E/A330L突变增强其ADCC效应，其序列如下所示：The TREM2 antibody PY314 from Pionyr Immunotherapeutics was used as a positive control antibody. The variable region amino acid sequence of PY314 is recorded in WO2019118513. It uses the IgG1 subtype and enhances its ADCC effect through the heavy chain constant region S239D/I332E/A330L mutation. Its sequence is shown below:
PY314重链序列，SEQ NO ID:88：
PY314 heavy chain sequence, SEQ NO ID: 88:
PY314轻链序列，SEQ NO ID:89：
PY314 light chain sequence, SEQ NO ID: 89:
将PY314的轻链和重链基因合成后分别亚克隆至pcDNA3.4表达载体，转染Expi CHO细胞，在37℃、5％CO2下培养后，收集细胞培养物，离心，并用Protein A亲和柱纯化，得到PY314抗体。The light chain and heavy chain genes of PY314 were synthesized and subcloned into the pcDNA3.4 expression vector, respectively, and transfected into Expi CHO cells. After culturing at 37°C and 5% CO2 , the cell culture was collected, centrifuged, and purified using a Protein A affinity column to obtain the PY314 antibody.
实施例2、全人重组抗体库筛选Example 2: Screening of fully human recombinant antibody library
通过固相与液相交叉筛选的方式，筛选全人重组抗体库(由三优生物制备)，通过胰酶洗脱的方式将针对人、小鼠、食蟹猴TREM2的特异性Fab抗体富集下来，通过ELISA检测不同输出集合的富集情况。The fully human recombinant antibody library (prepared by Sanyou Biotechnology) was screened by solid phase and liquid phase cross-screening, and the specific Fab antibodies against human, mouse, and crab-eating macaque TREM2 were enriched by trypsin elution, and the enrichment of different output sets was detected by ELISA.
全人重组抗体库淘选(Kingfisher法)：Panning of fully human recombinant antibody library (Kingfisher method):
(1)背景扣除：将全人重组抗体库噬菌体悬液(由三优生物制备)用5％ BSA稀释封闭，与Dynabeads(购自Invitrogen)共孵育，收集负筛孵育后的噬菌体。(1) Background subtraction: The phage suspension of the human recombinant antibody library (prepared by Sanyou Biotechnology) was diluted and blocked with 5% BSA, incubated with Dynabeads (purchased from Invitrogen), and the phages after negative screening incubation were collected.
(2)Dynabeads封闭及包被：采用Kingfisher磁珠筛选系统(购自Thermo scientific)对磁珠进行结合和清洗，将5％ BSA与磁珠孵育。(2) Dynabeads blocking and coating: The magnetic beads were bound and washed using the Kingfisher magnetic bead screening system (purchased from Thermo scientific), and 5% BSA was incubated with the magnetic beads.
(3)正筛：将负筛后收集的噬菌体悬液与生物素标记的TREM2蛋白(购自凯佧)包被且封闭后的Dynabeads孵育，并采用Kingfisher磁珠筛选系统进行结合和清洗。(3) Positive screening: The phage suspension collected after negative screening was incubated with Dynabeads coated and blocked with biotin-labeled TREM2 protein (purchased from Kaixia), and bound and washed using the Kingfisher magnetic bead screening system.
(4)洗脱：用Trypsin(购自上海生工)洗脱噬菌体。(4) Elution: Use Trypsin (purchased from Shanghai Biotechnology) to elute the phage.
(5)侵染库板：洗脱后噬菌体溶液与对数期SS320细胞充分混合后37℃静置孵育30min。涂布在2YT-Car+-Tet+平板上，37℃培养箱过夜培养。 (5) Infection of library plate: After elution, the phage solution was fully mixed with logarithmic phase SS320 cells and incubated at 37°C for 30 min. The cells were spread on 2YT-Car+-Tet+ plates and cultured in a 37°C incubator overnight.
(6)点Titer：将洗脱后噬菌体溶液用对数期SS320细胞10倍梯度稀释，37℃静置孵育30min，混匀后取2μL滴在平板上，37℃培养箱过夜培养。(6) Titer: The eluted phage solution was diluted 10-fold with logarithmic phase SS320 cells, incubated at 37°C for 30 min, mixed and dropped 2 μL on a plate, and cultured in a 37°C incubator overnight.
(7)统计：计算噬菌体输入、输出等，刮板制备输入的噬菌体并进行5轮筛选，每一轮筛选采用3倍梯度抗原递减的浓度筛选获得具有较高亲和力的抗体克隆。(7) Statistics: Calculate phage input, output, etc., prepare the input phage by scraping and perform 5 rounds of screening. Each round of screening uses a 3-fold gradient antigen decreasing concentration screening to obtain antibody clones with higher affinity.
全人重组抗体库淘选(免疫管法)：Selection of fully human recombinant antibody library (immunotube method):
(1)背景扣除：将全人重组抗体库噬菌体悬液(由三优生物制备)用5％ PBSM封闭。(1) Background subtraction: The phage suspension of the fully human recombinant antibody library (prepared by Sanyou Biotechnology) was blocked with 5% PBSM.
(2)免疫管包被封闭：将TREM2蛋白(购自凯佧)包被5％ PBSM封闭。(2) Immunotube coating and blocking: TREM2 protein (purchased from Kexia) was coated with 5% PBSM for blocking.
(3)正筛：将噬菌体悬液与蛋白包被且封闭后的免疫管孵育，并按照免疫管筛选系统方法进行结合和清洗。(3) Positive screening: incubate the phage suspension with the protein-coated and sealed immunotubes, and perform binding and washing according to the immunotube screening system method.
(4)洗脱：用Trypsin(购自上海生工)洗脱噬菌体。(4) Elution: Use Trypsin (purchased from Shanghai Biotechnology) to elute the phage.
(5)侵染库板：洗脱后噬菌体溶液与对数期SS320细胞充分混合后37℃静置孵育30min。涂布在2YT-Car+-Tet+平板上，37℃培养箱过夜培养。(5) Infection of library plate: After elution, the phage solution was fully mixed with logarithmic phase SS320 cells and incubated at 37°C for 30 min. The cells were spread on 2YT-Car+-Tet+ plates and cultured in a 37°C incubator overnight.
(6)点Titer：将洗脱后噬菌体溶液用对数期SS320细胞10倍梯度稀释，37℃静置孵育30min，混匀后取2μL滴在平板上，37℃培养箱过夜培养。(6) Titer: The eluted phage solution was diluted 10-fold with logarithmic phase SS320 cells, incubated at 37°C for 30 min, mixed and dropped 2 μL on a plate, and cultured in a 37°C incubator overnight.
(7)统计：计算噬菌体输入、输出等，刮板制备输入的噬菌体并进行3轮筛选，每一轮筛选采用3倍梯度抗原递减的浓度筛选获得具有较高亲和力的抗体克隆。(7) Statistics: Calculate phage input, output, etc., prepare the input phage by scraping and perform three rounds of screening. Each round of screening uses a 3-fold gradient antigen decreasing concentration screening to obtain antibody clones with higher affinity.
通过以下步骤用ELISA检测上述不同输出集合的富集情况：The enrichment of the above different output pools was tested by ELISA using the following steps:
(1)包被：将2μg/mL人、小鼠或食蟹猴TREM2(均购自凯佧)，按每孔30μL添加至ELISA板，4℃孵育过夜，PBST洗板3次。(1) Coating: 2 μg/mL human, mouse, or cynomolgus monkey TREM2 (all purchased from Kejia) was added to the ELISA plate at a rate of 30 μL per well, incubated overnight at 4°C, and the plate was washed three times with PBST.
(2)封闭：用5％ PBSM室温封闭1小时，PBST洗板3次。(2) Blocking: Block with 5% PBSM at room temperature for 1 hour, and wash the plate three times with PBST.
(3)一抗孵育：用5％ PBSM将上述不同输出集合的Fab或噬菌体表达上清按照5倍梯度进行稀释，加入稀释后上清，30μL/孔，室温孵育1小时，PBST洗板3次。(3) Primary antibody incubation: dilute the Fab or phage expression supernatant of the above different output sets with 5% PBSM in a 5-fold gradient, add the diluted supernatant, 30 μL/well, incubate at room temperature for 1 hour, and wash the plate 3 times with PBST.
(4)二抗孵育：Fab上清加入用PBS1:5000稀释的二抗Anti-mouse-Fab-HRP(购自Sino biological)，噬菌体加入用5％ PBSM 1:20000稀释的二抗Anti-M13-HRP(购自Sino biological)，30μL/孔，室温孵育1小时，PBST洗板6次。(4) Secondary antibody incubation: Add secondary antibody Anti-mouse-Fab-HRP (purchased from Sino biological) diluted 1:5000 in PBS to the Fab supernatant, and add secondary antibody Anti-M13-HRP (purchased from Sino biological) diluted 1:20000 in 5% PBSM to the phage, 30 μL/well, incubate at room temperature for 1 hour, and wash the plate 6 times with PBST.
(5)终止：加入30μL/孔的TMB(购自SurModics)室温显色5～10分钟，然后加入30μL/孔的终止液(2M硫酸)终止反应，酶标仪OD 450读取数据。(5) Termination: Add 30 μL/well of TMB (purchased from SurModics) to develop color at room temperature for 5 to 10 minutes, then add 30 μL/well of stop solution (2 M sulfuric acid) to terminate the reaction, and read the data using an OD 450 microplate reader.
选择在Fab水平或噬菌体水平上能够与人、小鼠、食蟹猴TREM2(均购自凯佧)交叉富集，或能够与人、食蟹猴TREM2交叉富集的输出集合。挑取第三轮、第四轮、第五轮筛选的输出集合的单克隆进行针对人、小鼠、食蟹猴TREM2的噬菌体ELISA初筛。初筛共挑取了3378个克隆，获得496个在蛋白ELISA水平具有人鼠猴或人猴交叉结合活性的阳性克隆，其中73个为序列独特的分子，制备噬菌体上清，通过ELISA进行针对人、小鼠、食蟹猴TREM2的结合活性检测，其中35个分子具有人鼠猴或人猴交叉结合活性，对这35个分子进行全长抗体的构建，采用IgG1亚型。其中部分分子进行重链恒定区S239D/I332E/A330L突变增强其ADCC效应，以HC表示，例如A10-HC表示经过S239D/I332E/A330L突变的A10分子。代表性的全人抗体库分子的序列见表1。Select output sets that can cross-enrich with human, mouse, and cynomolgus monkey TREM2 (all purchased from Kaixia) at the Fab level or phage level, or can cross-enrich with human and cynomolgus monkey TREM2. Pick monoclones from the output sets of the third, fourth, and fifth rounds of screening for phage ELISA screening of human, mouse, and cynomolgus monkey TREM2. A total of 3378 clones were picked in the initial screening, and 496 positive clones with human, mouse, monkey, or human-monkey cross-binding activity at the protein ELISA level were obtained, of which 73 were molecules with unique sequences, and phage supernatants were prepared. Binding activity against human, mouse, and cynomolgus monkey TREM2 was detected by ELISA, of which 35 molecules had human, mouse, monkey, or human-monkey cross-binding activity. Full-length antibodies were constructed for these 35 molecules, using the IgG1 subtype. Some of the molecules have heavy chain constant region S239D/I332E/A330L mutations to enhance their ADCC effect, represented by HC, for example, A10-HC represents the A10 molecule with S239D/I332E/A330L mutations. The sequences of representative fully human antibody library molecules are shown in Table 1.
全长抗体构建过程如下：将上述的35个Fab分子的基因以及人IgG1的重链和轻链恒定区(κ链)的基因分别克隆到pcDNA3.4表达载体中，通过测序分析获得序列正确的全长抗体表达质粒。按照ExpiFectamineTMCHO转染试剂(购自Thermo)的说明书，将纯化得到的所述全长抗体表达质粒转染入Expi CHO细胞中，在37℃、5％ CO2下培养后，离心收集上清，使用Protein A亲和层析纯化得到全长抗体。The full-length antibody construction process is as follows: the genes of the above-mentioned 35 Fab molecules and the genes of the heavy chain and light chain constant regions (κ chain) of human IgG1 are cloned into the pcDNA3.4 expression vector respectively, and the full-length antibody expression plasmid with the correct sequence is obtained by sequencing analysis. According to the instructions of ExpiFectamine TM CHO transfection reagent (purchased from Thermo), the purified full-length antibody expression plasmid is transfected into Expi CHO cells, and after culturing at 37°C and 5% CO2 , the supernatant is collected by centrifugation, and the full-length antibody is purified using Protein A affinity chromatography to obtain the full-length antibody.
表1全人抗体库分子的序列
Table 1 Sequences of fully human antibody library molecules
实施例3、全人抗体库分子的ELISA结合活性Example 3: ELISA binding activity of fully human antibody library molecules
按照如下步骤用ELISA检测全人抗体库分子与人、小鼠、食蟹猴TREM2的结合活性。The binding activity of the fully human antibody library molecules to human, mouse, and cynomolgus monkey TREM2 was detected by ELISA according to the following steps.
(1)包被：将2μg/mL人、小鼠或食蟹猴TREM2(均购自凯佧)，按每孔30μL添加至ELISA板，4℃孵育过夜，PBST洗板3次。(1) Coating: 2 μg/mL human, mouse, or cynomolgus monkey TREM2 (all purchased from Kejia) was added to the ELISA plate at a rate of 30 μL per well, incubated overnight at 4°C, and the plate was washed three times with PBST.
(2)封闭：用5％ PBSM室温封闭2小时，PBST洗板3次。 (2) Blocking: Block with 5% PBSM at room temperature for 2 hours, and wash the plate three times with PBST.
(3)一抗孵育：用1％ PBSM将待测抗体按照3倍梯度进行稀释，使终浓度分别为3、0.333、0.111、0.037、0.0123、0.00412、0.00137、0.00045μg/mL，加入稀释后抗体30μL/孔，室温孵育1小时，PBST洗板3次。(3) Primary antibody incubation: dilute the antibody to be tested with 1% PBSM in a 3-fold gradient to make the final concentrations 3, 0.333, 0.111, 0.037, 0.0123, 0.00412, 0.00137, and 0.00045 μg/mL, respectively. Add 30 μL/well of the diluted antibody and incubate at room temperature for 1 hour. Wash the plate 3 times with PBST.
(4)二抗孵育：加入用1％牛奶1:8000稀释的二抗Anti-human-Fc-IgG-HRP(购自abcam)，室温孵育1小时，PBST洗板6次。(4) Secondary antibody incubation: Add secondary antibody Anti-human-Fc-IgG-HRP (purchased from abcam) diluted 1:8000 with 1% milk, incubate at room temperature for 1 hour, and wash the plate 6 times with PBST.
(5)显色和终止：加入30μL/孔的TMB(购自SurModics)室温显色5～10分钟，然后加入30μL/孔的终止液(2M硫酸)终止反应，酶标仪OD 450读取数据。(5) Color development and termination: Add 30 μL/well of TMB (purchased from SurModics) for color development at room temperature for 5 to 10 minutes, then add 30 μL/well of stop solution (2 M sulfuric acid) to terminate the reaction, and read the data using an OD 450 microplate reader.
图1-1～图1-4(本申请全人抗体和阳性对照抗体与人TREM2蛋白的结合检测结果图)示出了全人抗体库分子与人TREM2蛋白的结合，图1-5～图1-8(本申请全人抗体和阳性对照抗体与小鼠TREM2蛋白的结合检测结果图)示出了全人抗体库分子与小鼠TREM2蛋白的结合，图1-9～图1-12(本申请全人抗体和阳性对照抗体与食蟹猴TREM2蛋白的结合检测结果图)示出了全人抗体库分子与食蟹猴TREM2蛋白的结合，可以看出，全人抗体库分子能够与人、鼠、猴TREM2蛋白交叉结合或与人、猴TREM2蛋白交叉结合。Figures 1-1 to 1-4 (the binding test results of the fully human antibodies and positive control antibodies of the present application with human TREM2 protein) show the binding of fully human antibody library molecules with human TREM2 protein, Figures 1-5 to 1-8 (the binding test results of the fully human antibodies and positive control antibodies of the present application with mouse TREM2 protein) show the binding of fully human antibody library molecules with mouse TREM2 protein, and Figures 1-9 to 1-12 (the binding test results of the fully human antibodies and positive control antibodies of the present application with cynomolgus monkey TREM2 protein) show the binding of fully human antibody library molecules with cynomolgus monkey TREM2 protein. It can be seen that the fully human antibody library molecules can cross-bind with human, mouse, and monkey TREM2 proteins or cross-bind with human and monkey TREM2 proteins.
实施例4、全人抗体库分子的FACS结合活性Example 4: FACS binding activity of fully human antibody library molecules
采用流式细胞仪分别检测全人抗体库分子与过表达人TREM2、小鼠TREM2、食蟹猴TREM2的CHO-K1细胞(均购自吉满基因)的结合活性，步骤如下。Flow cytometry was used to detect the binding activity of the fully human antibody library molecules with CHO-K1 cells overexpressing human TREM2, mouse TREM2, and cynomolgus monkey TREM2 (all purchased from Yoshiman Gene), as follows.
(1)按照1×105个/孔接种细胞，离心并用FACS缓冲液洗涤细胞。(1) Seed cells at 1×10 5 cells/well, centrifuge and wash the cells with FACS buffer.
(2)在细胞中加入100μL浓度梯度稀释(初始浓度50μg/mL，用FACS缓冲液(DPBS+1％ BSA)4倍稀释)后的全人抗体库分子，4℃孵育60min，用FACS缓冲液离心洗涤细胞2次。(2) Add 100 μL of fully human antibody library molecules after concentration gradient dilution (initial concentration 50 μg/mL, 4-fold dilution with FACS buffer (DPBS + 1% BSA)) to the cells, incubate at 4°C for 60 min, and wash the cells twice by centrifugation with FACS buffer.
(3)在细胞中加入100μL PE标记的anti-Human Fc(购自Abcam)，4℃孵育30min。(3) Add 100 μL PE-labeled anti-Human Fc (purchased from Abcam) to the cells and incubate at 4°C for 30 min.
(4)用FACS缓冲液重悬细胞，用流式细胞仪进行检测。(4) Resuspend the cells in FACS buffer and detect using flow cytometer.
图2-1～图2-3(本申请全人抗体和阳性对照抗体与人TREM2过表达细胞株的结检测结果图)示出了全人抗体库分子与人TREM2过表达细胞株的结合，图2-4～图2-8(本申请全人抗体和阳性对照抗体与小鼠TREM2过表达细胞株的结合检测结果图)示出了全人抗体库分子与小鼠TREM2过表达细胞株的结合，图2-9～图2-10(本申请全人抗体和阳性对照抗体与食蟹猴TREM2过表达细胞株的结合检测结果图)示出了全人抗体库分子与食蟹猴TREM2过表达细胞株的结合，其中，以PY314抗体作为阳性对照，IgG1同种型抗体作为阴性对照。可以看出，全人抗体库分子能够与人、鼠、猴TREM2过表达细胞株交叉结合或与人、猴TREM2过表达细胞株交叉结合。Figures 2-1 to 2-3 (the binding test results of the fully human antibody and positive control antibody of the present application with human TREM2 overexpressing cell lines) show the binding of the fully human antibody library molecules with human TREM2 overexpressing cell lines, Figures 2-4 to 2-8 (the binding test results of the fully human antibody and positive control antibody of the present application with mouse TREM2 overexpressing cell lines) show the binding of the fully human antibody library molecules with mouse TREM2 overexpressing cell lines, and Figures 2-9 to 2-10 (the binding test results of the fully human antibody and positive control antibody of the present application with cynomolgus monkey TREM2 overexpressing cell lines) show the binding of the fully human antibody library molecules with cynomolgus monkey TREM2 overexpressing cell lines, wherein the PY314 antibody is used as a positive control and the IgG1 isotype antibody is used as a negative control. It can be seen that the fully human antibody library molecules can cross-bind with human, mouse, and monkey TREM2 overexpressing cell lines or cross-bind with human and monkey TREM2 overexpressing cell lines.
实施例5：全人抗体库分子的ADCC效应Example 5: ADCC effect of fully human antibody library molecules
采用如下步骤检测全人抗体库分子介导ADCC效应的能力，以人PBMC细胞作为效应细胞，以过表达人TREM2的CHO-K1细胞(购自吉满基因)作为靶细胞。The following steps were used to detect the ability of the fully human antibody library molecules to mediate ADCC effects, using human PBMC cells as effector cells and CHO-K1 cells overexpressing human TREM2 (purchased from Yoshiman Gene) as target cells.
(1)按照1×104个细胞/孔、50μL/孔接种hu-TREM2-CHO-K1细胞，37℃孵育30min，同时设置靶细胞加效应细胞孔(仅加入靶细胞和PBMC细胞)、靶细胞非裂解孔(仅加入靶细胞)、靶细胞全裂解孔(仅加入靶细胞)。(1) hu-TREM2-CHO-K1 cells were inoculated at 1×10 4 cells/well and 50 μL/well and incubated at 37°C for 30 min. At the same time, target cell plus effector cell wells (only target cells and PBMC cells were added), target cell non-lysis wells (only target cells were added), and target cell full lysis wells (only target cells were added) were set.
(2)用含有1％ FBS的1840培养基对全人抗体库分子进行梯度稀释，按照50μL/孔加入到细胞中，使终浓度分别为2、0.2、0.02、0.002μg/mL，37℃孵育30min。(2) Use 1840 medium containing 1% FBS to perform gradient dilutions of the fully human antibody library molecules, add them to the cells at a rate of 50 μL/well to achieve final concentrations of 2, 0.2, 0.02, and 0.002 μg/mL, respectively, and incubate at 37°C for 30 min.
(3)按照5×105个细胞/孔、100μL/孔加入PBMC细胞(三优生物制备)，37℃孵育4h。(3) PBMC cells (prepared by Sanyou Biotechnology) were added at a density of 5×10 5 cells/well and 100 μL/well and incubated at 37°C for 4 h.
(4)在靶细胞全裂解孔中加入10μL 10％ Triton X-100，37℃孵育30min。(4) Add 10 μL 10% Triton X-100 to the target cell lysis well and incubate at 37°C for 30 min.
(5)离心后各孔取60μL上清液与LDH试剂等比例混合，室温孵育30min，在490nm处检测OD值。(5) After centrifugation, 60 μL of supernatant was taken from each well and mixed with LDH reagent in equal proportions. The mixture was incubated at room temperature for 30 min and the OD value was detected at 490 nm.
(6)按照如下公式计算裂解率：裂解率＝(样品孔OD值-靶细胞加效应细胞孔OD值)/(靶细胞全裂解孔OD值-靶细胞非裂解孔OD值)(6) Calculate the lysis rate according to the following formula: lysis rate = (OD value of sample well - OD value of target cell plus effector cell well) / (OD value of target cell fully lysed well - OD value of target cell non-lysed well)
图3-1～图3-3(本申请全人抗体的ADCC效应结果图)示出了全人抗体库分子的ADCC效应结果，可以看出，全人抗体库分子A10、A10-HC、A4、A4-HC、A11-HC具有较强的诱导ADCC效应的活性。Figures 3-1 to 3-3 (ADCC effect result diagrams of the fully human antibodies of the present application) show the ADCC effect results of the fully human antibody library molecules. It can be seen that the fully human antibody library molecules A10, A10-HC, A4, A4-HC, and A11-HC have strong activity in inducing ADCC effect.
实施例6：全人抗体库分子的体内药效研究Example 6: In vivo efficacy study of fully human antibody library molecules
对全人抗体库分子A10、A10-HC、A4、A4-HC、A47、F83-HC进行小鼠肿瘤模型体内药效研究，如前文所述，HC表示通过重链恒定区S239D/I332E/A330L突变增强ADCC效应的分子。为了更好地模拟在人体内的药效，使用转入了人TREM2基因的TREM2人源化C57BL/6小鼠(购自南模生物)。取雌性TREM2人源化小鼠，按2.5E6/只的数量皮下接种MC38小鼠结肠癌细胞，建立皮下荷瘤模型。至肿瘤生长至100mm3 左右，按照表2所示方案开始分组给药。研究过程中每天观察小鼠存活情况及一般状态，每周2次测量肿瘤体积和体重。The fully human antibody library molecules A10, A10-HC, A4, A4-HC, A47, and F83-HC were used for in vivo efficacy studies in mouse tumor models. As mentioned above, HC represents a molecule that enhances the ADCC effect through the heavy chain constant region S239D/I332E/A330L mutation. In order to better simulate the efficacy in humans, TREM2 humanized C57BL/6 mice (purchased from Shanghai Model Organisms) with human TREM2 genes were used. Female TREM2 humanized mice were subcutaneously inoculated with MC38 mouse colon cancer cells at a number of 2.5E6/mouse to establish a subcutaneous tumor-bearing model. When the tumor grew to 100 mm 3 Around 2:00 pm, grouping and drug administration began according to the scheme shown in Table 2. During the study, the survival and general condition of the mice were observed every day, and the tumor volume and body weight were measured twice a week.
至研究终点的肿瘤生长曲线图和体重变化趋势图分别如图4-1和图4-2所示，肿瘤生长抑制率(TGI)如表2所示，研究结果显示，全人抗体库分子均具有明显的抑瘤作用，其中F83-HC、A4、A47、A10-HC的抑瘤作用显著高于PY314，A10、A4-HC的抑瘤作用与PY314相当，实验过程中各组动物体重均无明显下降。The tumor growth curve and body weight change trend diagram to the end point of the study are shown in Figure 4-1 and Figure 4-2, respectively. The tumor growth inhibition rate (TGI) is shown in Table 2. The research results show that all molecules in the human antibody library have obvious tumor inhibition effects, among which the tumor inhibition effects of F83-HC, A4, A47, and A10-HC are significantly higher than that of PY314, and the tumor inhibition effects of A10 and A4-HC are equivalent to those of PY314. There was no significant decrease in the body weight of animals in each group during the experiment.
表2全人源库分子体内药效研究方案
Table 2 In vivo efficacy study plan for all human library molecules
将本申请的序列列于下表3中。The sequences of the present application are listed in Table 3 below.
表3
Table 3
为了描述和公开的目的，以引用的方式将所有的专利、专利申请和其它出版物在此明确地并入本申请。这些出版物仅因为它们的公开早于本申请的申请日而提供。所有关于这些文件的日期的声明或这些文件的内容的表述是基于申请者可得的信息，并且不构成任何关于这些文件的日期或这些文件的内容的正确性的承认。而且，在任何国家，在本中对这些出版物的任何引用并不构成关于该出版物成为本领域的公知常识的一部分的认可。For the purpose of description and disclosure, all patents, patent applications and other publications are expressly incorporated herein by reference. These publications are provided only because they are disclosed prior to the filing date of the present application. All statements about the dates of these documents or the representations of the contents of these documents are based on the information available to the applicant and do not constitute any admission of the correctness of the dates of these documents or the contents of these documents. Moreover, any reference to these publications in this document does not constitute an admission that the publications become part of the common general knowledge in the art in any country.
本领域技术人员将认识到，本申请的范围并不限于上文描述的各种具体实施方式和实施例，而是能够在不脱离本申请的精神的情况下，进行各种修改、替换、或重新组合，这都落入了本申请的保护范围内。 Those skilled in the art will recognize that the scope of the present application is not limited to the various specific implementations and examples described above, but that various modifications, replacements, or recombinations can be made without departing from the spirit of the present application, which all fall within the scope of protection of the present application.
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