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WO1994004191A1 - Traitement medical - Google Patents

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Publication number
WO1994004191A1
WO1994004191A1 PCT/GB1993/001716 GB9301716W WO9404191A1 WO 1994004191 A1 WO1994004191 A1 WO 1994004191A1 GB 9301716 W GB9301716 W GB 9301716W WO 9404191 A1 WO9404191 A1 WO 9404191A1
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WO
WIPO (PCT)
Prior art keywords
cells
compound
antibody
cell
cdr
Prior art date
Application number
PCT/GB1993/001716
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English (en)
Inventor
Nigel Stephen Courtenay-Luck
Original Assignee
Antisoma Limited
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Filing date
Publication date
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Publication of WO1994004191A1 publication Critical patent/WO1994004191A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
    • A61K51/1093—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody conjugates with carriers being antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/6425—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent the peptide or protein in the drug conjugate being a receptor, e.g. CD4, a cell surface antigen, i.e. not a peptide ligand targeting the antigen, or a cell surface determinant, i.e. a part of the surface of a cell
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6849—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2121/00—Preparations for use in therapy

Definitions

  • the present invention relates to the acquired immunodeficiency syndrome (AIDS) and infection with the human immunodeficiency virus type 1 (HIV-1).
  • AIDS acquired immunodeficiency syndrome
  • HIV-1 human immunodeficiency virus type 1
  • AIDS is thought to be caused by a virus, HIV-1, which damages the immune system by infecting and depleting the CD4 positive T helper/inducer subset of T cells and other CD4 positive cells of the immune system.
  • T cells are essential because they control the production of antibodies by the B cells, the maturation of cytotoxic T lymphocytes (killer T cells), the maturation and activity of macrophages and natural killer cells, and directly and indirectly, numerous other regulator and effector functions of the immune system. Infection with HTV-1 , therefore, leads to inadequate immune responses, thus permitting an HIV-1 infected individual to be afflicted by opportunistic infections, resulting in the weakening of the body and the eventual demise of the individual.
  • the CD4 antigen is the cell-surface receptor for HTV-1. Infection of CD4 positive cells is thought to occur through interaction between an epitope borne by HTV-1, known as gpl20, and the CD4 antigen located on the surface of T cells and also on other cells such as some macrophages or glial cells.
  • the virus After HTV binds to the CD4 molecule, the virus is internalised and uncoated. Once internalised, the genomic RNA is transcribed to DNA by the enzyme reverse transcriptase. The proviral DNA is then integrated into the host chromosomal DNA. The virus can then initiate the process of replication, shedding, and infection of further CD4 positive cells.
  • Previously most therapeutic systems against HIV infection and the treatment of AIDS have involved the development of drugs or vaccines that can eliminate the virus, or the production of molecules such as recombinant CD4 which can block the binding of the virus onto native CD4 receptors.
  • PCT/US90/03921 discloses the inhibition of HTV replication in human CD4 positive T cells by employing Pokeweed Antiviral Protein (PAP)-monoclonal antibody conjugates without detectable cytotoxicity.
  • PAP Pokeweed Antiviral Protein
  • the toxin acts only against the virus and not against the T-cells.
  • a first aspect of the present invention provides the use of a compound comprising a targeting portion to target the compound to a CD4 positive cell and a cytotoxic portion capable of destroying the CD4 positive cell when the compound is adjacent to said cell in the manufacture of a medicament for the treatment of infection by HIV viruses.
  • CD4 positive cell we mean any cell that expresses the protein molecule CD4.
  • the CD4 molecule is a receptor site located on the surface mainly of T-helper cells, but also of some macrophages or glial cells.
  • adjacent to we include the situation where the compound is internalised in the CD4 + cell.
  • the cytotoxic portion may comprise a toxic enzyme from a plant or bacterium.
  • Suitable bacterial toxins include diphtheria and Pseudomonas 3 toxin; suitable plant toxins include ricin and abrin. They may be produced by recombinant DNA techniques or by isolation from the natural source. DNA sequences encoding these proteins are publicly available from the GenBank, EMBL and Daresbury databases.
  • a modified form of a protein includes chemically modified forms as well as mutant forms created through genetic engineering. Chemical modifications include, for example, derivitization for the purpose of linking the moieties to each other, either directly or through a linking compound, by methods that are well known in the art of protein chemistry.
  • the means of linking the toxic moiety and the recognition moiety may comprise a heterobifunctional coupling reagent which ultimately contributes to formation of an intermolecular disulfide bond between the two moieties.
  • Suitable agents capable of reacting with thiol groups include N- hydroxysuccinimide ester of iodoacetic acid (NHIA) and N-succinimidyl-3- (2-pyridyldithio)propionate (SPDP).
  • the intermolecular disulfide may conveniently be formed between cysteines in each moiety which occur naturally or are inserted by genetic engineering.
  • the means of linking moieties may also use thioether linkages between heterobi unctional crosslinking reagents or specific low pH cleavable crosslinkers or specific protease cleavable linkers or other cleavable or noncleavable chemical linkages.
  • the means of linking moieties of the cytotoxic reagent may also comprise a peptidyl bond formed between moieties which are separately synthesised by standard peptide synthesis chemistry or recombinant means .
  • Possible chemical modifications of the protein moieties of the present invention also include derivatization with polyethylene glycol (PEG) to extend time of residence in the circulatory system and reduce immunogenicity, according to well known methods (see for examples, Lisi, P.J., Van Es, T., Abuchowski, A. et al (1982) Applied Biochem. 4, 19-33; Beauchamp, CO., Gonias, S.L., Menapace, D.P. et al (1982) Anal. Biochem. 131, 25-33; and Goodson, R.J. and Katre, N.V. (1990) Bio/Technology 8, 343-346).
  • PEG polyethylene glycol
  • Possible genetic engineering modifications of the proteins of the cytotoxic reagent include combination of the relevant functional domains of each into a single chain multi-functional biosynthetic protein expressed from a single gene derived by recombinant DNA techniques.
  • a single chain multi-functional biosynthetic protein expressed from a single gene derived by recombinant DNA techniques.
  • cytotoxic agents include chemotherapy agents, toxins, cytotoxic steroids, gelonin, and phospholipases.
  • Suitable chemotherapeutic agents are listed in the Table. TABLE: CHEMOTHERAPEUTIC AGENTS
  • Chemotherapeutic agents can be attached to proteins, including antibodies.
  • daunomycin and adriamvcin can be attached to antibodies using the method disclosed by Hurwitz et al (1975) "The covalent binding of daunomycin and adriamycin to antibodies with retention of both drug and antibody activities" Cancer Res. 35, 1175-1181.
  • Chlorambucil can be attached to proteins using the method of Ghose et al (1972) "Antibody as carrier of chlorambucil" Cancer 29, 1398-1412.
  • Methotrexate can be attached to proteins using the method of Chu et al (1979) J. Natl. Cancer Inst. 62, 79-82.
  • Diphtheria toxin can be attached to proteins using the method of Moolten et al (1972) J. Natl. Cancer Inst. 49, 1057-1062.
  • AZT and acyclovir to the protein using suitable chemical linking agents such as N-bromo-succinimide or those described by O'Sullivan et al (1979) Anal. Biochem. 100, 100-108.
  • a particularly preferred cytotoxic portion comprises a radioactive portion.
  • the radioactive portion may comprise any radioactive atom which emits enough energy to destroy the affected cell as well as neighbouring cells.
  • the radioactive portion is a short range radioisotope.
  • the radioactive portion may comprise iodine- 125, phosphorus-32, iodine-123, iodine-131, indium-I l l, rhenium-186, rhenium-188, yttrium-90 or astatine-211.
  • 1-123 is less preferred, as it is the only one which does not have a short range component.
  • Astatine-211 emits alpha particles.
  • the isotopes and density of radioactive atoms in the compound of the invention are such that a dose of more than 4000 cGy (preferably at least 6000, 8000 or 10000 cGy) is delivered to the cell and its organelles, particularly the nucleus.
  • the radioactive atom(s) may be inco ⁇ orated in the compound of the invention in known ways.
  • the targeting portion may be biosynthesized or may be synthesized by chemical amino acid synthesis using suitable amino acid precursors involving, for example, fluorine- 19 in place of hydrogen.
  • the targeting portion incorporates the radioactive portion.
  • the IODOGEN method described by Fraker et al (Biochem. Biophys. Res. Commun. (1978)) (which is incorporated herein by reference) can be used to incorporate iodine-125 or other radioiodines. Other methods are described in detail in "Monoclonal Antibodies in Immunoscintigraphy", J-F Chatal (CRC Press 1989).
  • Labels such as " m Tc, 123 I, Rh and ⁇ n In can be attached via cysteine residues in the targeting portion or an added cysteine, preferably terminal. Similarly, existing or added lysines may be used for addition of yttrium.
  • the compounds of the invention may alternatively be prepared by conjugating the cytotoxic moiety to the targeting portion by known methods (for example using N-bromo-succinimide or by direct condensation of the terminal carboxy group with the terminal amino group of two polypeptides) or by expressing a fused nucleotide sequence, when the cytotoxic portion is a polypeptide.
  • the targeting portion is an antibody specific for a CD4 positive cell.
  • Suitable MAbs include OKT4 and OKT4A. available from Ortho
  • anti-CD4 antibodies include A40 (mouse isotype IgGl) from AMS Biotechnology (UK) Ltd, Oxford; Clone T151 (mouse isotype IgG2a) from Boehringer Mannheim; MAB 1733 (IgM) and MAB 1732 (IgGl) from Chemicon International, London NW2; and YNB46.1.8 (rat isotype IgGl), B-B14 (mouse IgGl), B-F5 (mouse IgGl), B-Al (mouse IgG2a) all from Serotec, Kidlington, Oxford 0X5 1JE, UK.
  • the portion may be an entire antibody (usually, for convenience and specificity, a monoclonal antibody), a part or parts thereof (for example an F ab fragment, F(ab') 2 , dab or "minimum recognition unit") or a synthetic antibody or part thereof.
  • a compound comprising only part of an antibody may be advantageous by virtue of being less likely to undergo non-specific binding due to the F c part.
  • Suitable monoclonal antibodies to selected antigens may be prepared by known techniques, for example those disclosed in "Monoclonal Antibodies: A manual of techniques", H. Zola (CRC Press, 1988) and in “Monoclonal Hybridoma Antibodies: Techniques and Applications", J.G.R. Hurrell (CRC Press, 1982).
  • Bispecific antibodies may be prepared by cell fusion, by reassociation of monovalent fragments or by chemical cross-linlting of whole antibodies, with one part of the resulting bispecific antibody being directed to the CD4 molecule and the other to the cytotoxic portion.
  • the bispecific antibody can be administered bound to the cytotoxic portion or it can be administered first, followed by the cytotoxic portion. The former is preferred.
  • Methods for preparing bispecific antibodies are disclosed in Corvalan et al (1987 Cancer Immunol. Immunother. 24, 127- 132 and 133-137 and 138-143. Bispecific antibodies, chimeric antibodies and single chain antibodies are discussed generally by Williams in Tibtech, February 1988, Vol.
  • non- human antibodies can be "humanized” in known ways, for example by inserting the CDR regions of mouse antibodies into the framework of human antibodies. IgG class antibodies are preferred.
  • variable heavy (V H ) and variable light (V domains of the antibody are involved in antigen recognition, a fact first recognised by early protease digestion experiments. Further confirmation was found by "humanisation" of rodent antibodies. Variable domains of rodent origin may be fused to constant domains of human origin such that the resultant antibody retains the antigenic specificity of the rodent parented antibody (Morrison et al (1984) Proc. Natl. Acad. Sci. USA 81, 6851-6855).
  • variable domains that antigenic specificity is conferred by variable domains and is independent of the constant domains is known from experiments involving the bacterial expression of antibody fragments, all containing one or more variable domains.
  • variable domains include Fab-like molecules (Better et al (1988) Science 240, 1041); Fv molecules (Skerra et al (1988) Science 240, 1038); single-chain Fv (ScFv) molecules where the V H and V L partner domains are linked via a flexible oligopeptide (Bird et al (1988) Science 242, 423; Huston et al (1988) Proc. Natl. Acad. Sci.
  • ScFv molecules we mean molecules wherein the V H and V L partner domains are linked via a flexible oligopeptide.
  • the targeting portion may alternatively be any other compound that binds specifically onto the CD4 receptor.
  • the targeting portion may be the recombinant viral protein gpl20 or part thereof, which is itself non-toxic but can be attached to or internalised by CD4 positive cells.
  • the gpl20 protein may be purified from HIV particles but because of the difficulties associated with this approach it is usual to use gpl20 produced as a recombinant protein, for example from E. coli cells. Purification may be by affinity binding to CD4 or by ion exchange chromatography, both methods known in the art.
  • the gpl20 protein suitable for use in the invention is commercially available from Chemicon, London, UK.
  • Variants of gpl20 which bind CD4 are also useful in the practice of the invention.
  • variants we include insertions, deletions and substitutions, either conservative or non-conservative, where such changes may enhance binding to CD4, or at least do not substantially alter the binding of the polypeptide to CD4.
  • substitutions are intended combinations such as Gly, Ala; Val, lie, Leu; Asp, Glu; Asn, Gin; Ser, Thr; Lys, Arg; and Phe, Tyr.
  • Such variants may be made using the methods of protein engineering and site-directed mutagenesis as described below in relation to modifying antibodies.
  • the gpl20 cDNA sequence is disclosed in Fisher et al (1987) Nature 334, 444-447, Ratner et al (1987) AIDS Research & Human Retroviruses 3, 57- 69 and Burger et al (1991) Proc. Natl. Acad. Sci. USA 88, 1236-1240 and so the protein encoded thereby may be expressed in cells using methods known in the art.
  • gpl20 or antibodies or fragments thereof may be fused toxin polypeptides by recombinant DNA techniques.
  • the two portions are produced as a fusion compound by recombinant DNA techniques whereby a length of DNA comprises respective regions encoding the two portions either adjacent one another or separated by a region encoding a linker peptide which does not destroy the desired properties of the compound.
  • a benefit in making the compound of the invention using recombinant DNA techniques is that it enables a high degree of precision with which the two portions of the compound can be joined together.
  • the two portions of the compound may overlap wholly or partly.
  • the DNA is then expressed in a suitable host to produce a polypeptide comprising the compound of the invention.
  • the DNA encoding the polypeptide constituting the compound of the invention may be used in accordance with known techniques, appropriately modified in view of the teachings contained herein, to construct an expression vector, which is then used to transform an appropriate host cell for the expression and production of the polypeptide of the invention.
  • Such techniques include those disclosed in US Patent Nos.
  • DNA encoding the polypeptide constituting the compound of the invention may be joined to a wide variety of other DNA sequences for introduction into an appropriate host.
  • the companion DNA will depend upon the nature of the host, the manner of the introduction of the DNA into the host, and whether episomal maintenance or integration is desired.
  • the DNA is inserted into an expression vector, such as a plasmid, in proper orientation and correct reading frame for expression.
  • an expression vector such as a plasmid
  • the DNA may be linked to the appropriate transcriptional and translational regulatory control nucleotide sequences recognised by the desired host, although such controls are generally available in the expression vector.
  • the vector is then introduced into the host through standard techniques. Generally, not all of the hosts will be transformed by the vector. Therefore, it will be necessary to select for transformed host cells.
  • One selection technique involves inco ⁇ orating into the expression vector a DNA sequence, with any necessary control elements, that codes for a selectable trait in the transformed cell, such as antibiotic resistance.
  • the gene for such selectable trait can be on another vector, which is used to co-transform the desired host cell.
  • targeting the compound to a CD4 positive cell, we mean either that the targeting portion is capable of recognising and selectively binding to the CD4 receptor or that it may be specifically taken up by the CD4 receptor which is the intended target.
  • the compound is targeted primarily to human CD4 positive T cells, but also to some macrophages or glial cells.
  • a CDR-grafted antibody may be produced based on human framework regions and having an antigen binding site which recognises the CD4 antigen.
  • the CDR-grafted antibody has an affinity for the CD4 antigen similar to that of the murine MAb OKT4A.
  • a CDR-grafted antibody may be produced having at least one chain wherein the framework regions are predominantly derived from a first antibody (acceptor) and at least one CDR is derived from a second antibody (donor), the CDR-grafted antibody being capable of binding to the CD4 antigen.
  • the CDR-grafted chain may have two or all three CDRs derived from the donor antibody.
  • the or each CDR comprises a composite CDR comprising all the residues from the CDR and all the residues in the corresponding hypervariable region of the donor antibody.
  • At least one residue in the framework regions of the CDR- grafted chain has been altered so that it corresponds to the equivalent residue in the antibody, and the framework regions of the CDR-grafted chain are derived from a human antibody.
  • the framework regions of the CDR-grafted chain are derived from a human Ig heavy chain.
  • residue 35 in the heavy chain framework regions be altered so that it corresponds to the equivalent residue in the donor antibody.
  • At least one composite CDR comprising residues 26 to 35, 50 to 65 or 95 to 102 respectively is grafted onto the human framework. It will be appreciated in this case that residue 35 will already correspond to the equivalent residue in the donor antibody.
  • residues 23, 24 and 49 in such heavy chains correspond to the equivalent residues in the antibody. It is more preferred that residues 6, 23, 24, 48 and 49 in such heavy chains correspond to the donor antibody in equivalent residue positions. If desired, residues 71, 73 and 79 can also so correspond.
  • any one or any combination of residues 57, 58, 60, 88 and 91 may correspond to the equivalent residue in the donor antibody.
  • the heavy chain may be derived from the human KOL heavy chain. However, it may also be derived from the human NEWM or EU heavy chain.
  • the framework regions of the CDR-grafted chain may be derived from a human kappa or lambda light chain.
  • a human kappa or lambda light chain advantageously at least one composite CDR comprising residues 24 to 34, 50 to 56 or 89 to 97 respectively is grafted onto the human framework.
  • residue 49 also corresponds to the equivalent residue in the donor antibody.
  • residues 49 and 89 correspond to the equivalent residues in the donor antibody. It may also be desirable to select equivalent donor residues that form salt bridges.
  • the light chain is preferably derived from the human REI light chain. However, it may also be derived from the human EU light chain.
  • the CDR-grafted antibody comprises a light chain and a heavy chain, one or, preferably, both of which have been CDR-grafted in accordance with the principles set out above for the individual light and heavy chains.
  • the donor and acceptor residues may be identical at a particular position and thus no change of acceptor framework residue will be required.
  • the CDR-grafted antibody is a complete Ig, for example of isotype IgGj, or IgG 2 , IgG 3 or IgM.
  • one or more residues in the constant domains of the Ig may be altered in order to alter the effector functions of the constant domains.
  • the CDR-grafted antibody has an affinity for the CD4 antigen of between about lO ⁇ M "1 to about 10 12 .M _1 , more preferably at least lO ⁇ M 1 and most preferably the affinity is similar to that of MAb OKT4 or OKT4A.
  • the or each CDR is derived from a mammalian antibody and preferably is derived from a murine MAb.
  • the CDR-grafted antibody is produced by use of recombinant DNA technology.
  • a further method for producing a CDR-grafted antibody comprises providing a first DNA sequence, encoding a first antibody chain in which the framework regions are predominantly derived from a first antibody (acceptor) and at least one CDR is derived from a second antibody (acceptor), under the control of suitable upstream and downstream elements; transforming a host cell with the first DNA sequence; and culturing the transformed host cell so that a CDR-grafted antibody is produced.
  • the method further comprises: providing a second DNA sequence, encoding a second antibody chain complementary to the first chain, under the control of suitable upstream and downstream elements; and transforming the host cell with both the first and second DNA sequences.
  • the second DNA sequence encodes a second antibody chain in which the framework regions are predominantly derived from a first antibody (acceptor) and at least one CDR is derived from the second antibody (donor).
  • the first and second DNA sequences may be present on the same vector.
  • the sequences may be under the control of the same or different upstream and/or downstream genetic elements.
  • the first and second DNA sequences may be present on different vectors.
  • a nucleotide sequence may be formed which encodes an antibody chain in which the framework regions are predominantly derived from a first antibody (acceptor) and at least one CDR is derived from a second antibody (donor), the antibody chain being capable of forming a CDR- grafted antibody.
  • the CDR-grafted antibodies may be produced by a variety of techniques. with expression in transfected cells, such as yeast, insect, CHO or myeloma cells, being preferred. Most preferably, the host cell is a CHO host cell.
  • variable domain sequence of an antibody having the desired binding properties Suitable source cells for such DNA sequences include avian, mammalian or other vertebrate sources such as chickens, mice, rats and rabbits, and preferably mice.
  • the variable domain sequences (V H and V j J) may be determined from heavy and light chain cDNA, synthesized from the respective mRNA by techniques generally known to the art.
  • the hypervariable regions may then be determined using the Kabat method (Wu and Kabat, J. (1970) J. Exp. Med. 132, 211).
  • the CDRs may be determined by structural analysis using X-ray crystallography or molecular modelling techniques.
  • a composite CDR may then be defined as containing all the residues in one CDR and all the residues in the corresponding hypervariable region.
  • These composite CDRs along with certain select residues from the framework region are preferably transferred as the "antigen binding sites", while the remainder of the antibody, such as the heavy and light chain constant domains and remaining framework regions, may be based on human antibodies of different classes. Constant domains may be selected to have desired effector functions appropriate to the intended use of the antibody so constructed.
  • human IgG isotypes, IgG ⁇ and IgG 3 are effective for complement fixation and cell mediated lysis.
  • other isotypes such as IgG 2 and IgG 4 , or other classes, such as IgM and IgE, may be more suitable.
  • Human constant domain DNA sequences preferably in conjunction with their variable domain framework bases can be prepared in accordance with well-known procedures. An example of this is CAMPATH 1H available from The Wellcome Foundation Ltd.
  • CDR-grafted antibodies which contain select alterations to the human-like framework region (in other words, outside of the CDRs of the variable domains), resulting in a CDR-grafted antibody with satisfactory binding affinity.
  • binding affinity is preferably from about l ⁇ .M *1 to about lO ⁇ .M "1 and is more preferably at least about lO ⁇ M "1 .
  • Most preferably the binding affinity is about equal to that of murine MAb OKT4A.
  • V H and/or V L gene segments may be altered by mutagenesis.
  • nucleotides coding for amino acid residues or sequences contained in the Fc portion or other areas of the antibody may be altered in like manner (see, for example, PO7US89/00297).
  • Exemplary techniques include the addition, deletion or nonconservative substitution of a limited number of various nucleotides or the conservative substitution of many nucleotides, provided that the proper reading frame is maintained.
  • Substitutions, deletions, insertions or any subcombination may be used to arrive at a final construct. Since there are 64 possible codon sequences but only twenty known amino acids, the genetic code is degenerate in the sense that different codons may yield the same amino acid. Thus there is at least one codon for each amino acid, ie each codon yields a single amino acid and no other. It will be apparent that during translation, the proper reading frame must be maintained in order to obtain the proper amino acid sequence in the polypeptide ultimately produced.
  • exemplary techniques include oligonucleotide-mediated site-directed mutagenesis and the polymerase chain reaction.
  • Oligonucleotide site-directed mutagenesis in essence involves hybridizing an oligonucleotide coding for a desired mutation with a single strand of DNA containing the region to be mutated and using the single strand as a template for extension of the oligonucleotide to produce a strand containing the mutation. This technique, in various forms, is described in Zoller and Smith (1982) Nuc. Acids. Res. 10, 6487.
  • PCR Polymerase chain reaction
  • the oligonucleotides can inco ⁇ orate sequence alterations if desired.
  • the polymerase chain reaction technique is described in Mullis and Fuloona (1987) Meth. Enz. 155, 335. Examples of mutagenesis using PCR are described in Ho et al (1989) Gene 77, 51.
  • the nucleotide sequences capable of ultimately expressing the desired CDR-grafted antibodies, can be formed from a variety of different poly nucleotides (genomic DNA, cDNA, RNA or synthetic oligonucleotides). At present, it is preferred that the polynucleotide sequence comprises a fusion of cDNA and genomic DNA.
  • the polynucleotide sequence may encode various Ig components (eg V, J, D, and C domains). They may be constructed by a variety of different techniques. Joining appropriate genomic and cDNA sequences is presently the most common method of production, but cDNA sequences may also be utilized (see EP-A-0 239 400).
  • DNA constructs can be made that express anti-CD4 antibodies or antibody fragments, either humanised or non-humanised, or that express gpl20, or that express a polypeptide toxin for use in the invention or that express a fusion between an anti-CD4 antibody or antibody fragment and toxin or between gpl20 and toxin.
  • vectors and methods disclosed herein are suitable for use in host cells over a wide range of prokaryotic and eukaryotic organisms.
  • prokaryotes are preferred for cloning of DNA sequences for constructing the vectors.
  • E. coli DH5 ⁇ is particularly useful. This example is, of course, intended to be illustrative rather than limiting.
  • Prokaryotes may also be used for expression.
  • the aforementioned E. coli strains, bacilli such as Bacillus subtilus, and other enterobacteriaceae, such as Salmonella typhimurium or Serratia marcescens, and various Pseudomonas species may be used.
  • plasmid vectors containing replicon and control sequences which are derived from species compatible with the host cell are used in connection with these hosts.
  • the vector ordinarily carries a replication site as well as genetic sequences which are capable of providing phenotypic selection in transformed cells.
  • E. coli is typically transformed using one of the many derivatives of pBR322, a plasmid derived from an E. coli species (Bolivar et al (1977) Gene 2, 95).
  • pBR322 contains genes for ampicillin and tetracycline resistance and thus provides easy means for identifying transformed cells.
  • the pBR322 plasmid, its descendents or other microbial plasmids may also contain, or be modified to contain, promoters which can be used by the microbial organism for the expression of recombinant proteins.
  • promoters commonly used in recombinant DNA construction include lactose promoter systems (Goedell et al (1979) Nature 281, 544) and tryptophan (tip) promoter systems (Goedell et al (1980) Nuc. Acids. Res. 8, 4057 and EP-A-0 036 776).
  • eukaryotic microbes such as yeast cultures
  • Saccharomyces cerevisiae or common baker's yeast
  • Saccharomyces cerevisiae is the most commonly used among eukaryotic microorganisms, although a number of other strains are commonly available.
  • the plasmid YRp7 for example, is commonly used (Tschemper et al (1980) Gene 19, 157).
  • This plasmid already contains the tipl gene which provides a selection marker for a mutant strain of yeast lacking the ability to grow in tryptophan, for example ATCC No. 44076 or PEP4-1 (Jones (1977) Genetics 85, 12).
  • the presence of the tipl lesion as a characteristic of the yeast host cell genome then provides an effective environment for detecting transformation by growth in the absence of tryptophan.
  • Suitable promoting sequences in yeast vectors include the promoters for the genes encoding production of 3- ⁇ hosphoglycerate kinase or other glycolytic enzymes, such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate iso erase, 3-phosphoglycerate mutase, pyruvate kinase, triosephosphate isomerase, phosphoglucose isomerase and glucokinase.
  • 3- ⁇ hosphoglycerate kinase or other glycolytic enzymes such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate iso erase, 3-phosphoglycerate mut
  • the termination sequences associated with these genes are also ligated into the expression vector 3' of the sequence desired to be expressed to provide polyadenylation of the mRNA and termination.
  • Other promoters which have the additional advantage of transcription controlled by growth conditions are the promoter regions for alcohol dehydrogenase 2, isocytochrome C, acid phosphatase, degradative enzymes associated with nitrogen metabolism and the aforementioned glyceraldehyde-3-phosphate dehydrogenase, an enzyme responsible for maltose and galactose utilisation (Holland et al (1978) Biochemistry YJ, 4900). Any plasmid vector containing a yeast compatible promoter, origin of replication and termination sequences is suitable.
  • cultures of cells derived from multicellular organisms may also be used as hosts.
  • any such cell culture is workable, whether from a vertebrate or an invertebrate organism.
  • vertebrate cells have become a routine procedure in recent years.
  • useful host cell lines are VERO, HeLa, Chinese hamster ovary (CHO), W138, BHK, COS-7,
  • Expression vectors for such cells may include (if necessary) an appropriate origin of replication, as well as a promoter located in front of the gene to be expressed, along with any necessary ribosome binding sites, RNA splice sites, polyadenylation sites and transcriptional terminator sequences.
  • the control functions on the expression vectors are often provided by viral material.
  • commonly used promoters are derived from human Cytomegalovirus (HCMV), Polyoma virus, Adenovirus 2 and, most frequently, Simian Virus 40 (SV40).
  • the early and late promoters of SV40 virus are particularly useful because both are obtained easily from the virus as a fragment which also contains the SV40 viral origin of replication (Fiers et al (1978) Nature 273, 113). Further, it is also possible, and often desirable, to utilise promoter or control sequences normally associated with the desired gene sequence, provided such control sequences are compatible with the host cell system.
  • An origin of replication may be provided either by construction of the vector to include an exogenous origin, such as may be derived from SV40 or other viral (eg Polyoma virus, Adeno virus, VSV or BPV) source, or may be provided by the host cell chromosomal replication mechanism. If the vector is integrated into the host cell chromosome, the latter is often sufficient.
  • an exogenous origin such as may be derived from SV40 or other viral (eg Polyoma virus, Adeno virus, VSV or BPV) source, or may be provided by the host cell chromosomal replication mechanism. If the vector is integrated into the host cell chromosome, the latter is often sufficient.
  • the vectors containing the DNA segments of interest can be transferred into the host cell by well-known methods, which vary depending on the type of cellular host. For example, calcium chloride transfection is commonly utilised for prokaryotic cells, whereas calcium phosphate treatment, lipofection or electroporation may be used for other cellular hosts.
  • the CDR-grafted antibodies can be purified according to standard procedures of the art, including ammonium sulphate precipitation, affinity columns, column chromatography and gel electrophoresis. Binding affinities of the constructs so expressed may be ascertained by techniques known to the art.
  • Substantially pure CDR-grafted antibodies of at least 90 to 95% homogeneity are preferred, and 98 to 99% or more homogeneity is most preferred for pharmaceutical uses.
  • the CDR-grafted antibodies or recombinant gpl20 protein or fractions thereof can be iyophilised for storage and reconstituted in a suitable carrier prior to use.
  • This technique has been shown to be effective with conventional immunoglobulins and art-known lyophilisation and reconstitution techniques can be employed. It will be appreciated by those skilled in the art that lyophilisation and reconstitution can lead to varying degrees of antibody activity loss (eg with conventional immunoglobulins, IgM antibodies tend to have greater activity loss than IgG antibodies) and that use levels may have to be adjusted to compensate.
  • a further aspect of the invention provides a method of treating HIV (human immunodeficiency virus) infection in a patient comprising administering to the patient a compound comprising a targeting portion to target the compound to a CD4 positive cell and a cytotoxic portion capable of destroying the CD4 positive cell when the compound is adjacent to or internalised by the said cell.
  • HIV human immunodeficiency virus
  • the compounds of the present invention will be utilised in purified form together with pharmacologically appropriate carriers.
  • these carriers include aqueous or alcoholic/aqueous solutions. emulsions or suspensions, including saline and buffered media.
  • Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride and lactated Ringer's.
  • Suitable physiologically acceptable adjuvants if necessary to keep the complex in suspension, may be chosen from thickeners such as carboxymethylcellulose, polyvinylpyrrolidone, gelatin and alginates.
  • Intravenous vehicles include fluid and nutrient replenishes and electrolyte replenishers, such as those based on Ringer's dextrose. Preservatives and other additives, such as antimicrobials, antioxidants, chelating agents and inert gases, can also be present.
  • the route of administration of pharmaceutical compositions according to the invention may be any of those commonly known to those of ordinary skill in the art in standard sterile, non-pyrogenic formulations of diluents and carriers.
  • the administration can be by any appropriate mode, usually parenterally, including intravenously, intramuscularly, intraperitoneally, intra-arterially, intrathecally or also, appropriately, by direct infusion with a catheter.
  • the dosage and frequency of administration will depend on the age, sex and condition of the patient, concurrent administration of other drugs, counter indications and other parameters to be taken into account by the clinician. If needed, because the compound of the invention may be immunogenic, cyclosporin or some other immunosuppressant can be administered to provide a longer period for treatment but usually this will not be necessary.
  • the targeting portion of the compound is a monoclonal antibody or part thereof or recombinant gpl20 protein or part thereof which is radioactively labelled.
  • the virus will have no host to infect. HIV, like many other viruses, is a parasite and needs a host for its survival and proliferation, and all host cells which bind the virus through the CD4 receptor and allow infection by HTV are eliminated.
  • radioactive atoms are delivered to be cytotoxic to all CD4 positive cells without damaging other normal marrow or epithelial cells.
  • a dose of up to 100 mg of radiolabelled antibody may be suitable on any one occasion.
  • a short range radioisotope is used that can kill a cell only when internalised.
  • isotopes include iodine-125 or other auger emitting or alpha particle emitting radioisotopes such as astatine.
  • HTV particles in the absence of CD4 positive cells, will have no host to infect and therefore will be rapidly cleared from the body of the infected host. At that point, repopulation by new and uninfected CD4 positive cells will be allowed to commence.
  • the patient will have previously received antiviral chemotherapy such as AZT or DDI to inactive the HIV viruses during their release from infected CD4 positive cells, and broad spectrum antibiotics and antifungal agents for prophylaxis during the period of low CD4 counts.
  • antiviral chemotherapy such as AZT or DDI to inactive the HIV viruses during their release from infected CD4 positive cells, and broad spectrum antibiotics and antifungal agents for prophylaxis during the period of low CD4 counts.
  • Example 1 Protocol for radiolabelling anti-CD4 with isotopes of iodine using the IODO-GEN" method
  • the anti-CD4 monoclonal antibody which is internalised, once bound onto CD4 positive cells is radiolabelled.
  • the hybridoma secreting anti-CD4 may be grown in media containing 10% foetal calf serum or in serum free medium. The supernatant may then be separated by centrifugation from the cells and the antibody purified by protein-A affinity chromatography.
  • the antibody is radiolabelled to a high specific activity but without loss of immunoreactivity as described.
  • the MAb is labelled with 123 I, 131 I or 125 I using the iodogen technique.
  • the radiolabelled MAb is purified by gel filtration on a 20 ml Sephadex G-50 column previously treated with HSA.
  • IODO-GEN TM (Pierce Chemical Company) tubes are prepared by dissolving l,3,4,6,-Tetrachloro-3,6-diphenylglycouril in chloroform at a concentration of 1 mg.ml "1 . Aliquots of 50 ⁇ l (50 ⁇ g IODO-GEN TM ) are dispensed into polypropylene cryo-tubes and the chloroform evaporated to dryness. These tubes may be stored desiccated at -20°C until use.
  • Approximately 1 mg of MAb is dissolved in approximately 0.4 ml of a suitable buffer eg PBS.
  • the IODO-GEN TM tubes are equilibrated to room temperature before use.
  • the MAb solution is placed into the IODO-GEN TM tube along with the required isotope of iodine (approximately 1-10 ⁇ l in aqueous solution).
  • the reaction is allowed to proceed at room temperature for 5-15 min with occasional shaking. Following incubation the reaction mixture is removed and eluted through a Sephadex TM G50 column as previously described for DTPA conjugation to MAb.
  • the separation of MAb-iodine from free iodine is achieved in a similar manner to that described for separation of MAb- m In from free lu In.
  • DTPA is diethylenetriaminepenta acetic acid.
  • Example 2 Protocol for radiolabelling gp!20 with isotopes of iodine using the IODO-GEN" method
  • the recombinant gpl20 is labelled with 123 I, 131 I or 125 I using the iodogen technique.
  • the radiolabelled gpl20 is purified by gel filtration on a 20 ml Sephadex G-50 column previously treated with HSA.
  • IODO-GEN TM (Pierce Chemical Company) tubes are prepared by dissolving l,3,4,6,-Tetrachloro-3,6-diphenylglycouril in chloroform at a concentration of 1 mg.ml "1 . Aliquots of 50 ⁇ l (50 ⁇ g IODO-GEN TM ) are dispensed into polypropylene cryo-tubes and the chloroform evaporated to dryness. These tubes may be stored desiccated at -20°C until use.
  • gpl20 Approximately 1 mg of gpl20 is dissolved in approximately 0.4 ml of a suitable buffer eg PBS.
  • a suitable buffer eg PBS e.g PBS.
  • the IODO-GEN TM tubes are equilibrated to room temperature before use.
  • the gpl20 solution is placed into the IODO- GEN TM tube along with the required isotope of iodine (approximately 1-10 ⁇ l in aqueous solution).
  • the reaction is allowed to proceed at room temperature for 5-15 min with occasional shaking. Following incubation the reaction mixture is removed and eluted through a Sephadex TM G50 column as previously described for DTPA conjugation to gpl20.
  • the separation of gpl20-iodine from free iodine is achieved in a similar manner to that described for separation of MAb- m In from free m In.
  • Example 3 Example of radiolabelling a protein with m In
  • the protein is conjugated to the chelating agent DTPA, using the bicyclic anhydride method (Hnatowich, D. J. et al (1983) J. Immunol. Methods 65, 147-157).
  • Free DTPA is separated from DTPA-MAb by Sephadex G- 50 gel-filtration using 0.1 M sodium acetate as elution buffer.
  • ul In citrate is added to the DTPA conjugate and allowed to react for 15-30 mins. Purification is as for the iodinated protein.
  • the Sephadex TM column should be washed three times with its own volume of phosphate buffered saline (PBS) before elution of the MAb/DTPA reaction mixture. Fractions, eluted from the column in PBS, are collected and analysed for the presence of protein by measuring abso ⁇ tion in a spectrophotometer at 280 nm. Fractions which are positive for the presence of protein are then pooled. Care should be taken, throughout the procedure, to avoid contact of all materials with metallic ions since chelation of metal ions by DTPA will result in a reduced labelling efficiency when subsequently labelling with In.
  • PBS phosphate buffered saline
  • ⁇ n In a solution of ⁇ n In chloride is adjusted to pH 6.0-7.5 with sodium citrate to give a final activity of approximately 250 MBq.ml "1 .
  • An appropriate volume of the m ln solution is then added to the required amount of MAb-DTPA solution to give an appropriate specific activity of ⁇ n In:Moab.
  • the exact radiolabelling efficiency for a particular MAb-DTPA conjugate should be determined by experimentation.
  • n ⁇ In solution the reaction is allowed to proceed for 10 min at room temperature.
  • MAb- associated m In is separated from free m In by use of a Sephadex TM G50 column as described for DTPA coupling.
  • Fractions are collected from the column and measured for the presence of radioactivity and normally results in the appearance of two distinct radioactive peaks from the column.
  • the first peak correlates with the protein peak, as determined by spectrophotometry, whilst the second peak represents free or unbound m In. Fractions comprising the first peak are collected and pooled for subsequent use.
  • Example 4 Labelling protein with Yttrium-90
  • Yttrium-90 in 0.04 M hydrochloric acid is complexed using 0.5 sodium acetate.
  • Benzyl-DTPA-conjugated protein is added to the complex.
  • EDTA is added to react with free 90 Y. Separation is as for iodinated protein.
  • a patient who is HTV positive and may have AIDS is given intravenously, for a systemic effect, and (if appropriate) intrathecally, for the treatment of CNS infection, radioactive protein.
  • the patient is receiving systemic antiviral, antibiotic and antifungal therapy and housed in a sterile room for the period of CD4 lymphopenia.

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Abstract

Procédé de traitement d'une infection par HIV (virus de l'immunodéficience humaine) chez un malade et comprenant l'administration d'un composé comportant une partie de ciblage servant à cibler ledit composé vers une cellule positive CD4 et une partie cytotoxique pouvant détruire la cellule positive CD4 quand ledit composé est contigu à ladite cellule ou intégré par cette dernière. Les particules de HIV, en l'absence de cellules positives CD4, ne possèdent aucun hôte à infecter et, de ce fait, sont rapidement évacuées du corps de l'hôte infecté.
PCT/GB1993/001716 1992-08-13 1993-08-13 Traitement medical WO1994004191A1 (fr)

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE19809785A1 (de) * 1998-03-08 1999-09-09 Bergter Radioimmunpharmakon zur Behandlung der HIV-1-Infektion
WO1999054954A2 (fr) * 1998-04-15 1999-10-28 Wolfgang Bergter Substances pharmaceutiques radio-immunologiques a base de cd4 destinees au traitement de l'infection par le vih
WO1999055385A2 (fr) * 1998-04-28 1999-11-04 Wolfgang Bergter Produits radioimmunopharmaceutiques pour le traitement de l'hepatite c
DE19826307A1 (de) * 1998-03-08 1999-12-16 Wolfgang Bergter CD4-Radioimmunpharmaka zur Behandlung der HIV-Infektion
CN109251893A (zh) * 2018-09-18 2019-01-22 深圳市宝迪生物工程有限公司 一种离体胎盘造血干细胞的制备方法
US11126212B2 (en) 2017-02-15 2021-09-21 Systemex Energies Inc. Power control device

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1988009181A2 (fr) * 1987-05-29 1988-12-01 Tanox Biosystems, Inc. Anticorps monoclonaux neutralisant le hiv-1
WO1989006967A1 (fr) * 1988-02-03 1989-08-10 Xoma Corporation Immunosuppresion a l'aide de compositions d'immunotoxines de cellules t anti-pan
WO1990001035A1 (fr) * 1988-07-23 1990-02-08 The United States Of America, Represented By The Secretary, United States Department Of Commerce Agent cytotoxique pour le traitement d'infections virales specifiques
EP0381763A1 (fr) * 1988-06-10 1990-08-16 Shuzo Matsushita Anticorps modifie par une toxine
WO1990012868A1 (fr) * 1989-04-25 1990-11-01 Tanox Biosystems, Inc. Anticorps specifiques contre le domaine de liaison de cd4 du virus d'immunodeficience humaine
WO1991000360A1 (fr) * 1989-06-29 1991-01-10 Medarex, Inc. Reactifs bispecifiques pour le traitement du sida
WO1991001145A1 (fr) * 1989-07-25 1991-02-07 Regents Of The University Of Minnesota Conjugues de proteines antivirales pokeweed - anticorps monoclonaux

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1988009181A2 (fr) * 1987-05-29 1988-12-01 Tanox Biosystems, Inc. Anticorps monoclonaux neutralisant le hiv-1
WO1989006967A1 (fr) * 1988-02-03 1989-08-10 Xoma Corporation Immunosuppresion a l'aide de compositions d'immunotoxines de cellules t anti-pan
EP0381763A1 (fr) * 1988-06-10 1990-08-16 Shuzo Matsushita Anticorps modifie par une toxine
WO1990001035A1 (fr) * 1988-07-23 1990-02-08 The United States Of America, Represented By The Secretary, United States Department Of Commerce Agent cytotoxique pour le traitement d'infections virales specifiques
WO1990012868A1 (fr) * 1989-04-25 1990-11-01 Tanox Biosystems, Inc. Anticorps specifiques contre le domaine de liaison de cd4 du virus d'immunodeficience humaine
WO1991000360A1 (fr) * 1989-06-29 1991-01-10 Medarex, Inc. Reactifs bispecifiques pour le traitement du sida
WO1991001145A1 (fr) * 1989-07-25 1991-02-07 Regents Of The University Of Minnesota Conjugues de proteines antivirales pokeweed - anticorps monoclonaux

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
A. ADACHI ET AL: "GENERATION AND CHARACTERIZATION OF THE HIV VIRUS TYPE 1 MUTANTS.", ARCH. VIROL, vol. 117, no. 1-2, 1991, pages 45 - 58 *
BIOLOGICAL ABSTRACTS, vol. 92, Philadelphia, PA, US; abstract no. 4097 *
CHEMICAL ABSTRACTS, vol. 112, no. 5, 29 January 1990, Columbus, Ohio, US; abstract no. 30364v *
F. M. UCKUN ET AL.: "CELL-TYPE-SPECIFIC CYTOTOXICITY OF ANTI-CD4 AND ANTI-CD8 RICIN IMMUNOTOXINS AGAINST HUMAN ALLOREACTIVE T- CELL CLONES.", BLOOD, vol. 74, no. 7, 1989, pages 2445 - 2454 *
P. ASHORN ET AL.: "ELIMINATION OF INFECTIOUS HIV VIRUS FROM HUMAN T-CELL CULTURES BY SYNERGISTIC ACTION OF CD4-PSEUDOMONAS EXOTOXIN AND REVERSE TRANSCRIPTASE INHIBITORS.", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA, vol. 87, November 1990 (1990-11-01), WASHINGTON US, pages 8889 - 8893, XP055140624, DOI: doi:10.1073/pnas.87.22.8889 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE19809785A1 (de) * 1998-03-08 1999-09-09 Bergter Radioimmunpharmakon zur Behandlung der HIV-1-Infektion
WO1999045969A2 (fr) * 1998-03-08 1999-09-16 Wolfgang Bergter Medicament radioimmunnologique destine au traitement de l'infection a vih-1
DE19826307A1 (de) * 1998-03-08 1999-12-16 Wolfgang Bergter CD4-Radioimmunpharmaka zur Behandlung der HIV-Infektion
DE19809785C2 (de) * 1998-03-08 2000-02-10 Wolfgang Bergter Radioimmunpharmakon zur Behandlung der HIV-1-Infektion
WO1999045969A3 (fr) * 1998-03-08 2000-02-17 Wolfgang Bergter Medicament radioimmunnologique destine au traitement de l'infection a vih-1
WO1999054954A2 (fr) * 1998-04-15 1999-10-28 Wolfgang Bergter Substances pharmaceutiques radio-immunologiques a base de cd4 destinees au traitement de l'infection par le vih
WO1999054954A3 (fr) * 1998-04-15 2000-01-20 Wolfgang Bergter Substances pharmaceutiques radio-immunologiques a base de cd4 destinees au traitement de l'infection par le vih
WO1999055385A2 (fr) * 1998-04-28 1999-11-04 Wolfgang Bergter Produits radioimmunopharmaceutiques pour le traitement de l'hepatite c
WO1999055385A3 (fr) * 1998-04-28 2000-03-16 Wolfgang Bergter Produits radioimmunopharmaceutiques pour le traitement de l'hepatite c
US11126212B2 (en) 2017-02-15 2021-09-21 Systemex Energies Inc. Power control device
CN109251893A (zh) * 2018-09-18 2019-01-22 深圳市宝迪生物工程有限公司 一种离体胎盘造血干细胞的制备方法

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