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lllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllll · 2016. 6. 21. ·...
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United States Patent [191 Queen et al. lllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllll US005585089A [11] Patent Number: [45] Date of Patent: 5,585,089 Dec. 17, 1996 [54] HUMANIZED HVIMUNOGLOBULINS [75] Inventors: Cary L. Queen, Los Altos; Harold E. Selick, Belmont, both of Calif. [73] Assignee: Protein Design Labs, Inc., Mountain View, Calif. [21] Appl. No.: 477,728 [22] Filed: Jun. 7, 1995 Related US. Application Data [63] Continuation of Ser. No. 634,278, Dec. 19, 1990, Pat. No. 5,530,101, which is a continuation-in-part of Ser. No. 590, 274, Sep. 28, 1990, abandoned, and Ser. No. 310,252, Feb. 13, 1989, abandoned, which is a continuation-in-part of Ser. No. 290,975, Dec. 28, 1988, abandoned. [51] Int. Cl.6 ....................... .. C07K 16/18; A6lK 39/395 [52] US. Cl. ................................... .. 424/133.1; 530/387.3; 530/388.22; 424/1431 [58] Field of Search .......................... .. 530/387.3, 388.22; 424/1331, 143.1 [56] References Cited U.S. PATENT DOCUMENTS 4,578,335 3/1986 Urdal et a1. ........................... .. 530/351 4,816,397 3/1989 Boss et a1. .. .... .. 435/68 4,816,565 3/1989 Honjo et a1. . . . . . . . . . .. 435/691 4,816,567 3/1989 Cabilly et a1. .. ...... .. 530/387 4,845,198 7/1989 Urdal et a1. ........................ .. 530/387.3 4,867,973 9/1989 Goers et a1. . 5,198,359 3/1993 Taniguchi et a1. ................. .. 435/2523 5,225,539 7/1993 Winter ................................ .. 530/387.3 FOREIGN PATENT DOCUMENTS 0171496 2/1986 European Pat. Off. ...... .. C12N 15/00 0173494 3/1986 European Pat. O?“. ...... .. C12N 15/00 0184187 6/1986 European Pat. Off. ...... .. C12N 15/00 0256654 7/1987 European Pat. O?f. . 0239400 9/1987 European Pat. Off. . 0266663 6/1988 European Pat. 01f. ...... .. C12N 15/00 2188941 10/1987 United Kingdom .... .. C12N 5/00 86/05513 9/1986 WIPO .............. .. . C12N 15/00 87/02671 5/1987 WIPO C07H 15/12 89/01783 3/1989 WIPO ........................ .. A61K 39/395 OTHER PUBLICATIONS Riechmann et al. Nature vol. 332 24, Mar. 1988 p. 323. Foote, Nova Acta Leopoldina 1989. vol. 61 (269) 103. Amit et al. Science vol. 233 1986 p. 747. Groves et al. vol. 6, 1987, p. 71. Better et al., “Escherichia coli Secretion of an Active Chimeric Antibody Fragment”, Science 240:1041-1043 (1988). Bird et al., “Single-Chain Antigen—Binding Proteins”, Sci ence 242:423-426 (1988). Boulianne et al., “Production of functional chimeric mouse/ human antibody,” Nature 312:643-646 (1984). Carter et al., “Humanization of an anti-plSSmYR2 antibody for human cancer therapy,” Proc. Natl. Acad. Sci. 89:4285-4289 (1992). Chothia, C. and A. M. Lesk, “Canonical Structures for the Hypervariable Regions of Immunoglobulins”, J. Mol. Biol. 196:901-917 (1987). C0 et al., “Humanized antibodies for antiviral therapy,” Proc. Natl. Acad. Sci. USA 88:2869—2873 (1991). C0 et al., “Chimeric and Humanized Antibodies with Speci ?city for the CD33 Antigen,” J. of Immunol. 148(4):1149-1154 (1992). Daugherty et al., “Polymerase chain reaction facilitates the cloning, CDR-grafting, and rapid expression of a murine monoclonal antibody directed against the CD18 component of leukocyte integrins,” Nuc. Acids Res. 19:2471-2476 (1991). Ellison et al., “The nucleotide sequence of a human immu noglobulin C(garnma)1 gene”, Nucleic Acids Res. 10:4071—(1982). Farrar, J., “The biochemistry, biology, and role of interleu kin-2 in the induction of cytotoxic T cell and antibody forming B cell receptors,” Immunol. Rev. 63:129-166 (1982). Foote et al., “Antibody framework residues aifecting the conformation of hypervariable loops,” J. Mol. Biol. 224:487—499 (1992). Gorman et al., “Reshaping a therapeutic CD4 antibody,” Proc. Natl. Acad. Sci. 88:4181-4185 (1991). Greene et al., “Growth of Human T Lymphocytes: An Analysis of Interleukin 2 and Its Cellular receptor”, in Progress in Hematology XIV, E. Brown, ed., Grune and Statton, New York (1986) pp. 283-301. Hale et al., “Remission Induction in Non-Hodgkin Lym phoma with Reshaped Human Monoclonal Antibody CAMPATH—1H”, Lancet Dec. 17, 1988, pp. 1394-1399. Hieter et al., “Cloned Human and Mouse Kappa Immuno globulin Constant and .1 Region Genes Conserve Homology in Functional Segments”, Cell 22:197-—207 (1980). (List continued on next page.) Primary Examiner—Lila Feisee Attorney, Agent, or Firm—Townsend and Townsend and Crew LLP [57] ABSTRACT Novel methods for producing, and compositions of human ized immunoglobulins having one or more complementarity determining regions (CDR’ s) and possible additional amino acids from a donor immunoglobulin and a framework region from an accepting human immunoglobulin are provided. Each humanized immunoglobulin chain will usually com~ prise, in addition to the CDR’s, amino acids from the donor immunoglobulin framework that are, e.g., capable of inter acting with the CDR’s to eifect binding a?inity, such as one or more amino acids which are immediately adjacent to a CDR in the donor immunoglobulin or those within about 3 A as predicted by molecular modeling. The heavy and light chains may each be designed by using any one or all of various position criteria. When combined into an intact antibody, the humanized immunoglobulins of the present invention will be substantially non-immunogenic in humans and retain substantially the same a?inity as the donor immunoglobulin to the antigen, such as a protein or other compound containing an epitope. 11 Claims, 55 Drawing Sheets
Transcript of lllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllll · 2016. 6. 21. ·...
United States Patent [191 Queen et al.
lllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllll US005585089A
[11] Patent Number:
[45] Date of Patent:
5,585,089 Dec. 17, 1996
[54] HUMANIZED HVIMUNOGLOBULINS
[75] Inventors: Cary L. Queen, Los Altos; Harold E. Selick, Belmont, both of Calif.
[73] Assignee: Protein Design Labs, Inc., Mountain View, Calif.
[21] Appl. No.: 477,728
[22] Filed: Jun. 7, 1995
Related US. Application Data
[63] Continuation of Ser. No. 634,278, Dec. 19, 1990, Pat. No. 5,530,101, which is a continuation-in-part of Ser. No. 590, 274, Sep. 28, 1990, abandoned, and Ser. No. 310,252, Feb. 13, 1989, abandoned, which is a continuation-in-part of Ser. No. 290,975, Dec. 28, 1988, abandoned.
[51] Int. Cl.6 ....................... .. C07K 16/18; A6lK 39/395
[52] US. Cl. ................................... .. 424/133.1; 530/387.3; 530/388.22; 424/1431
[58] Field of Search .......................... .. 530/387.3, 388.22; 424/1331, 143.1
[56] References Cited
U.S. PATENT DOCUMENTS
4,578,335 3/1986 Urdal et a1. ........................... .. 530/351
4,816,397 3/1989 Boss et a1. .. .... .. 435/68
4,816,565 3/1989 Honjo et a1. . . . . . . . . . .. 435/691
4,816,567 3/1989 Cabilly et a1. .. ...... .. 530/387
4,845,198 7/1989 Urdal et a1. ........................ .. 530/387.3
4,867,973 9/1989 Goers et a1. . 5,198,359 3/1993 Taniguchi et a1. ................. .. 435/2523
5,225,539 7/1993 Winter ................................ .. 530/387.3
FOREIGN PATENT DOCUMENTS
0171496 2/1986 European Pat. Off. ...... .. C12N 15/00 0173494 3/1986 European Pat. O?“. ...... .. C12N 15/00 0184187 6/1986 European Pat. Off. ...... .. C12N 15/00 0256654 7/1987 European Pat. O?f. . 0239400 9/1987 European Pat. Off. . 0266663 6/1988 European Pat. 01f. ...... .. C12N 15/00 2188941 10/1987 United Kingdom .... .. C12N 5/00 86/05513 9/1986 WIPO .............. .. . C12N 15/00
87/02671 5/1987 WIPO C07H 15/12 89/01783 3/1989 WIPO ........................ .. A61K 39/395
OTHER PUBLICATIONS
Riechmann et al. Nature vol. 332 24, Mar. 1988 p. 323. Foote, Nova Acta Leopoldina 1989. vol. 61 (269) 103. Amit et al. Science vol. 233 1986 p. 747. Groves et al. vol. 6, 1987, p. 71. Better et al., “Escherichia coli Secretion of an Active Chimeric Antibody Fragment”, Science 240:1041-1043 (1988). Bird et al., “Single-Chain Antigen—Binding Proteins”, Sci ence 242:423-426 (1988). Boulianne et al., “Production of functional chimeric mouse/ human antibody,” Nature 312:643-646 (1984). Carter et al., “Humanization of an anti-plSSmYR2 antibody for human cancer therapy,” Proc. Natl. Acad. Sci. 89:4285-4289 (1992). Chothia, C. and A. M. Lesk, “Canonical Structures for the Hypervariable Regions of Immunoglobulins”, J. Mol. Biol. 196:901-917 (1987).
C0 et al., “Humanized antibodies for antiviral therapy,” Proc. Natl. Acad. Sci. USA 88:2869—2873 (1991). C0 et al., “Chimeric and Humanized Antibodies with Speci ?city for the CD33 Antigen,” J. of Immunol. 148(4):1149-1154 (1992). Daugherty et al., “Polymerase chain reaction facilitates the cloning, CDR-grafting, and rapid expression of a murine monoclonal antibody directed against the CD18 component of leukocyte integrins,” Nuc. Acids Res. 19:2471-2476 (1991). Ellison et al., “The nucleotide sequence of a human immu noglobulin C(garnma)1 gene”, Nucleic Acids Res. 10:4071—(1982). Farrar, J., “The biochemistry, biology, and role of interleu kin-2 in the induction of cytotoxic T cell and antibody forming B cell receptors,” Immunol. Rev. 63:129-166 (1982). Foote et al., “Antibody framework residues aifecting the conformation of hypervariable loops,” J. Mol. Biol. 224:487—499 (1992). Gorman et al., “Reshaping a therapeutic CD4 antibody,” Proc. Natl. Acad. Sci. 88:4181-4185 (1991). Greene et al., “Growth of Human T Lymphocytes: An Analysis of Interleukin 2 and Its Cellular receptor”, in Progress in Hematology XIV, E. Brown, ed., Grune and Statton, New York (1986) pp. 283-301. Hale et al., “Remission Induction in Non-Hodgkin Lym phoma with Reshaped Human Monoclonal Antibody CAMPATH—1H”, Lancet Dec. 17, 1988, pp. 1394-1399. Hieter et al., “Cloned Human and Mouse Kappa Immuno globulin Constant and .1 Region Genes Conserve Homology in Functional Segments”, Cell 22:197-—207 (1980).
(List continued on next page.)
Primary Examiner—Lila Feisee Attorney, Agent, or Firm—Townsend and Townsend and Crew LLP
[57] ABSTRACT
Novel methods for producing, and compositions of human ized immunoglobulins having one or more complementarity determining regions (CDR’ s) and possible additional amino acids from a donor immunoglobulin and a framework region from an accepting human immunoglobulin are provided. Each humanized immunoglobulin chain will usually com~ prise, in addition to the CDR’s, amino acids from the donor immunoglobulin framework that are, e.g., capable of inter acting with the CDR’s to eifect binding a?inity, such as one or more amino acids which are immediately adjacent to a CDR in the donor immunoglobulin or those within about 3 A as predicted by molecular modeling. The heavy and light chains may each be designed by using any one or all of various position criteria. When combined into an intact antibody, the humanized immunoglobulins of the present invention will be substantially non-immunogenic in humans and retain substantially the same a?inity as the donor immunoglobulin to the antigen, such as a protein or other compound containing an epitope.
11 Claims, 55 Drawing Sheets
5,585,089 Page 2
OTHER PUBLICATIONS
Huston et al., “Protein engineering of antibody binding sites: Recovery of speci?c activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli", Proc. Natl. Acad. Sci. USA. 85:5879-5883 (1988). in Progress in Hematology XIV, E. Brown, ed., Grune and Statton, New York (1986) p. 283. Jones et al., “Replacing the complementarity-determining regions in a human antibody with those from a mouse”, Nature 32l:522—525 (1986). Kettleborough et al., “Humanization of a mouse monoclonal antibody by CDR-grafting: the importance of framework residues on loop conformation,” Protein Engineering 41773-783 (1991). Kirkman et al., JoumalofExpt. Med. vol. 1621358 Jul. 1985. Leonard et al., “The human receptor for T-cell growth factor,” J. Biol. Chem. 260:1872-1880 (1985). Liu et al., “Expression of mouse::human immunoglobulin heavy-chain cDNA in lymphoid cells”, Gene 54:33—40 (1987). Maeda et al., “Construction of reshaped human antibodies with HIV—I1€lllI3llZlIlg activity”, Hum. Antibod. Hybrid. 2:124—l34 (1991). Morrison et al., “Chimeric human antibody molecules: Mouse antigen—binding domains with human constant region domains,” Proc. Natl. Acad. Sci. 81:6851-6859 (1984). Morrison, S. L., “Transfectomas Provide Novel Chimeric Antibodies,” Science 229:1202—1207 (1985). Neuberger et al., “A hapten-speci?c chimeric lgE antibody with human physiological effector function,” Nature 314:268—270 (1985). Queen et al., “A humanized antibody that binds to the interleukin 2 receptor,” Proc. Natl. Acad. Sci. USA 86:10029—10033 (1989). Riechmann et al., “Reshaping human antibodies for therapy”, Nature 332:323—327 (1988). Routledge et al., “A humanized monovalent CD3 antibody which can activate homologous complement,” Eur. J. Immu nol. 21: 27l7~2725 (1991).
Sahagan et al., “A Genetically Engineered Murine/Human Chimeric Antibody Retains Speci?city for Human Tumor —Associated Antigen”, J. Immunol. 137:1066—1074 (1986). Shalaby et al., “Development of humanized bispeci?c anti bodies reactive with cytotoxic lymphocytes and tumor cells overexpressing the HER2 protooncogene,” J. Exp. Med. 175:217-225 (1992). Sharon et al., “Expression of a V ,,C K chimaen'c protein in mouse myeloma cells”, Nature 309:364-367 (1984). Shearman et al., “Construction, expression and character ization of humanized antibodies directed against the human ot/B T cell receptor,” J. Immunol. l47(l2):4366—4373 (1991). Takeda et al., “Construction of chimaeric processed immu noglobulin genes containing mouse variable and human constant region sequences”, Nature 3141452-454 (1985). Tan et al., “A Human-Mouse Chimeric lmmunoglobulin Gene with a Human Variable Region is Expressed in Mouse Myeloma Cells”, J. Immunol. 135:3564-3567 (1985). Tempest et al., “Reshaping a human monoclonal antibody to inhibit human respiratory syncytial virus infection in vivo,” Bio/Technology 91226-271 (1991). Uchiyama et al., “A monoclonal antibody (anti—Tac) reac tive with activated and functionally mature human T-cells,” J. Immunol. l26:l393—l397 (1981). Verhoeyen et al., “Reshaping Human Antibodies: Grafting an Antilysozyme Activity”, Science 23911534-1536 (1988). Vitteta et al., “Redesigning Nature’s Poisons to Create Anti-Tumor Reagents,” Science 238:1098—ll04 (1987). Waldmann, T. A., “The Structure, Function, and Expression of Interleukin—2 Receptors on Normal and Malignant Lym phocytes,” Science 232:727—732 (1986). Woodle et al., “Humanized OKT3 antibodies: successful transfer of immune modulating properties and idiotype expression,” J. Immunol. l48:2756-—2763 (1992). Junghans et al., Cancer Res., 50:1495~l502 (1990). Kupiec—Weglinski et al., Proc. Natl. Acad. Sci., 8312624 (1986).
US. Patent Dec. 17, 1996 Sheet 1 of 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet 2 of 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet 3 of 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet v4 0f 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet 5 of 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet 7 of 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet 8 0f 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet 9 of 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet 10 of 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet 14 of 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet 15 of 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet 16 of 55 5,585,089
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US. Patent Dec. 17, 1996 Sheet '17 of 55 5,585,089
US. Patent Dec. 17, 1996 Sheet 18 of 55 5,585,089
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