Antibody drug conjugates

Abstract

The present invention relates to anti-FGFR2 and FGFR4 antibodies, antibody fragments, antibody drug conjugates, and their uses for the treatment of cancer.

1 . An antibody drug conjugate of the formula
   Ab-(L-(D) m ) n      
       or a pharmaceutically acceptable salt thereof; wherein 
       Ab is an antibody or antigen binding fragment thereof that specifically binds to both human 
       FGFR2 and FGFR4; 
       L is a linker; 
       D is a drug moiety; 
       m is an integer from 1 to 8; and 
       n is an integer from 1 to 10. 
     
     
         2 . The antibody drug conjugate of  claim 1 , wherein said m is 1. 
     
     
         3 . The antibody drug conjugate of  claim 1 , wherein said n is 3 or 4. 
     
     
         4 . The antibody drug conjugate of  claim 1 , wherein said antibody or antigen binding fragment specifically binds to all isoforms of human FGFR2. 
     
     
         5 . The antibody drug conjugate of  claim 1 , wherein said antibody or antigen binding fragment specifically binds to an epitope on human FGFR2 comprising amino acid residues 176 (Lys) and 210 (Arg) of SEQ ID NO:137. 
     
     
         6 . The antibody drug conjugate of  claim 1 , wherein said antibody or antigen binding fragment recognizes amino acid residues 173 (Asn), 174 (Thr), 175 (Val), 176 (Lys), 178 (Arg), 208 (Lys), 209 (Val), 210 (Arg), 212 (Gln), 213 (His), 217 (Ile), and 219 (Glu) of SEQ ID NO:137. 
     
     
         7 . The antibody drug conjugate of  claim 1 , wherein said antibody or antigen binding fragment specifically binds to an epitope of human FGFR2 comprising SEQ ID NO:136 or SEQ ID NO:141. 
     
     
         8 . The antibody drug conjugate of  claim 1 , wherein said antibody or antigen binding fragment specifically binds to an epitope of human FGFR2 consisting of SEQ ID NO:136 or SEQ ID NO:141. 
     
     
         9 . The antibody drug conjugate of  claim 1 , wherein said antibody or antigen binding fragment specifically binds to D1 and D2 domains of human FGFR4. 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . (canceled) 
     
     
         14 . (canceled) 
     
     
         15 . (canceled) 
     
     
         16 . (canceled) 
     
     
         17 . (canceled) 
     
     
         18 . The antibody drug conjugate of  claim 1 , wherein said antibody is a human antibody. 
     
     
         19 . The antibody drug conjugate of  claim 1 , wherein said antibody is a monoclonal antibody. 
     
     
         20 . The antibody drug conjugate of  claim 1 , wherein said linker is selected from the group consisting of a cleavable linker, a non-cleavable linker, a hydrophilic linker, a procharged linker and a dicarboxylic acid based linker. 
     
     
         21 . The antibody drug conjugate of  claim 20 , wherein the linker is derived from a cross-linking reagent selected from the group consisting of N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), N-succinimidyl 4-(2-pyridyldithio)pentanoate (SPP), N-succinimidyl 4-(2-pyridyldithio)butanoate (SPDB), N-succinimidyl-4-(2-pyridyldithio)-2-sulfo-butanoate (sulfo-SPDB), N-succinimidyl iodoacetate (SIA), N-succinimidyl(4-iodoacetyl)aminobenzoate (SIAB), maleimide PEG NHS, N-succinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (SMCC), N-sulfosuccinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (sulfo-SMCC) or 2,5-dioxopyrrolidin-1-yl 17-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-5,8,11,14-tetraoxo-4,7,10,13-tetraazaheptadecan-1-oate (CX1-1). 
     
     
         22 . The antibody drug conjugate of  claim 21 , wherein said linker is derived from the cross-linking reagent N-succinimidyl 4-(maleimidomethyl)cyclohexanecarboxylate (SMCC). 
     
     
         23 . The antibody drug conjugate of  claim 1 , wherein said drug moiety is selected from a group consisting of a V-ATPase inhibitor, a pro-apoptotic agent, a Bcl2 inhibitor, an MCL1 inhibitor, a HSP90 inhibitor, an IAP inhibitor, an mTor inhibitor, a microtubule stabilizer, a microtubule destabilizer, an auristatin, a dolastatin, a maytansinoid, a MetAP (methionine aminopeptidase), an inhibitor of nuclear export of proteins CRM1, a DPPIV inhibitor, proteasome inhibitors, inhibitors of phosphoryl transfer reactions in mitochondria, a protein synthesis inhibitor, a kinase inhibitor, a CDK2 inhibitor, a CDK9 inhibitor, a kinesin inhibitor, an HDAC inhibitor, a DNA damaging agent, a DNA alkylating agent, a DNA intercalator, a DNA minor groove binder and a DHFR inhibitor. 
     
     
         24 . The antibody drug conjugate of  claim 23 , wherein the cytotoxic agent is a maytansinoid. 
     
     
         25 . The antibody drug conjugate of  claim 24 , wherein the maytansinoid is N(2′)-deacetyl-N(2′)-(3-mercapto-1-oxopropyl)-maytansine (DM1) or N(2′)-deacetyl-N2-(4-mercapto-4-methyl-1-oxopentyl)-maytansine (DM4). 
     
     
         26 . The antibody drug conjugate of  claim 1  having the following formula: 
       
         
           
           
               
               
           
         
       
       wherein Ab is an antibody or antigen binding fragment thereof comprising a heavy chain CDR1 of SEQ ID NO: 1, a heavy chain CDR2 of SEQ ID NO: 2, a heavy chain CDR3 of SEQ ID NO: 3, and a light chain CDR1 of SEQ ID NO: 11, a light chain CDR2 of SEQ ID NO: 12, a light chain CDR3 of SEQ ID NO: 13, wherein the CDR is defined in accordance with the Kabat definition; and
 n is 1 to 10; or a pharmaceutically acceptable salt thereof. 
 
     
     
         27 . A pharmaceutical composition comprising the antibody drug conjugate of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         28 . The pharmaceutical composition of  claim 27  wherein said composition is prepared as a lyophilisate. 
     
     
         29 . The pharmaceutical composition of  claim 28 , wherein said lyophilisate comprises said antibody drug conjugate, sodium succinate, and polysorbate 20. 
     
     
         30 . A method of treating an FGFR2 positive or FGFR4 positive cancer in a patient in need thereof, comprising administering to said patient the antibody drug conjugate of  claim 1 . 
     
     
         31 . The method of  claim 30 , wherein said cancer is selected from the group consisting of gastric cancer, breast cancer, alveolar rhabdomyosarcoma, liver cancer, adrenal cancer, lung cancer, colon cancer and endometrial cancer. 
     
     
         32 . The method of  claim 30  further comprising administering to said patient a tyrosine kinase inhibitor, an IAP inhibitor, a Bcl2 inhibitor, an MCL1 inhibitor, or another FGFR2 inhibitor. 
     
     
         33 . The method of  claim 32 , wherein said another FGFR2 inhibitor is 3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-{6-[4-(4-ethyl-piperazin-1-yl)-phenylamino]-pyrimidin-4-yl}-1-methyl-urea. 
     
     
         34 . (canceled) 
     
     
         35 . (canceled) 
     
     
         36 . An antibody or antigen binding fragment thereof that specifically binds to FGFR2 and FGFR4. 
     
     
         37 . The antibody or antigen binding fragment of  claim 36 , wherein said antibody or antigen binding fragment specifically binds to an epitope on human FGFR2 comprising amino acid residues 176 (Lys) and 210 (Arg) of SEQ ID NO:137. 
     
     
         38 . (canceled) 
     
     
         39 . (canceled) 
     
     
         40 . The antibody or antigen binding fragment of  claim 37 , wherein said antibody or antigen binding fragment specifically binds to D1 and D2 domains of human FGFR4. 
     
     
         41 . (canceled) 
     
     
         42 . The antibody or antigen binding fragment of  claim 36  comprising a heavy chain variable region that comprises: (a) a VH CDR1 of SEQ ID NO: 1, (b) a VH CDR2 of SEQ ID NO: 2, and (c) a VH CDR3 of SEQ ID NO: 3, wherein the CDR is defined in accordance with the Kabat definition. 
     
     
         43 . The antibody or antigen binding fragment of  claim 42  further comprising a light chain variable region that comprises: (a) a VL CDR1 of SEQ ID NO: 11, (b) a VL CDR2 of SEQ ID NO: 12, and a VL CDR3 of SEQ ID NO: 13, wherein the CDR is defined in accordance with the Kabat definition. 
     
     
         44 . (canceled) 
     
     
         45 . (canceled) 
     
     
         46 . (canceled) 
     
     
         47 . (canceled) 
     
     
         48 . The antibody or antigen binding fragment of  claim 36 , wherein said antibody is a human antibody. 
     
     
         49 . The antibody or antigen binding fragment of  claim 36 , wherein said antibody is a monoclonal antibody. 
     
     
         50 . The antibody or antigen binding fragment of  claim 36 , wherein said antibody or antigen binding fragment is a single chain antibody (scFv). 
     
     
         51 . A nucleic acid that encodes the antibody or antigen binding fragment of  claim 36 . 
     
     
         52 . A vector comprising the nucleic acid of  claim 51 . 
     
     
         53 . A host cell comprising the vector according to  claim 52 . 
     
     
         54 . A process for producing an antibody or antigen binding fragment comprising cultivating the host cell of  claim 53  and recovering the antibody from the culture. 
     
     
         55 . A process for producing an anti-FGFR2 and FGFR4 antibody drug conjugate comprising:
 (a) chemically linking SMCC to a drug moiety DM-1;   (b) conjugating said linker-drug to the antibody recovered from the cell culture of  claim 54 ; and   (c) purifying the antibody drug conjugate.   
     
     
         56 . The antibody drug conjugate made according to  claim 55  having an average DAR, measured with a UV spectrophotometer, about 3.5. 
     
     
         57 . A diagnostic reagent comprising the antibody or antigen binding fragment thereof of  claim 36 , which is labeled. 
     
     
         58 . The diagnostic reagent of  claim 57 , wherein the label is selected from the group consisting of a radiolabel, a fluorophore, a chromophore, an imaging agent, and a metal ion. 
     
     
         59 . The antibody drug conjugate of  claim 1 , wherein said antibody or antigen binding fragment thereof comprises a heavy chain variable region that comprises: (a) a VH CDR1 of SEQ ID NO: 81, (b) a VH CDR2 of SEQ ID NO: 82, and (c) a VH CDR3 of SEQ ID NO: 83, wherein the CDR is defined in accordance with the Kabat definition. 
     
     
         60 . The antibody drug conjugate of  claim 59 , wherein said antibody or antigen binding fragment thereof further comprises a light chain variable region that comprises: (a) a VL CDR1 of SEQ ID NO: 91, (b) a VL CDR2 of SEQ ID NO: 92, and (c) a VL CDR3 of SEQ ID NO: 93, wherein the CDR is defined in accordance with the Kabat definition. 
     
     
         61 . The antibody drug conjugate of  claim 60 , wherein said antibody or antigen binding fragment thereof comprises a VH region of SEQ ID NO: 87 and a VL region of SEQ ID NO: 97. 
     
     
         62 . The antibody drug conjugate of  claim 61 , wherein said antibody consists of a heavy chain of SEQ ID NO: 89 and a light chain of SEQ ID NO: 99. 
     
     
         63 . The antibody of  claim 36 , wherein said antibody or antigen binding fragment thereof comprises a heavy chain variable region that comprises: (a) a VH CDR1 of SEQ ID NO: 81, (b) a VH CDR2 of SEQ ID NO: 82, and (c) a VH CDR3 of SEQ ID NO: 83, wherein the CDR is defined in accordance with the Kabat definition. 
     
     
         64 . The antibody of  claim 63 , wherein said antibody or antigen binding fragment thereof further comprises a light chain variable region that comprises: (a) a VL CDR1 of SEQ ID NO: 91, (b) a VL CDR2 of SEQ ID NO: 92, and (c) a VL CDR3 of SEQ ID NO: 93, wherein the CDR is defined in accordance with the Kabat definition. 
     
     
         65 . The antibody of  claim 64 , wherein said antibody or antigen binding fragment thereof comprises a VH region of SEQ ID NO: 87 and a VL region of SEQ ID NO: 97. 
     
     
         66 . The antibody of  claim 65 , wherein said antibody consists of a heavy chain of SEQ ID NO: 89 and a light chain of SEQ ID NO: 99.