US2006233814A1PendingUtilityA1

Elimination of heterogeneous or mixed cell population in tumors

Assignee: IMMUNOGEN INCPriority: Apr 15, 2005Filed: Apr 14, 2006Published: Oct 19, 2006
Est. expiryApr 15, 2025(expired)· nominal 20-yr term from priority
A61K 31/58C07K 2317/24A61P 37/00A61P 33/00C07D 498/18C07K 16/3046A61K 2039/505A61K 47/6849A61P 31/04A61K 31/5383A61P 31/12A61P 35/00A61K 47/68033A61K 47/6803A61K 38/19A61K 39/395
64
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Claims

Abstract

Methods of killing or inhibiting tumors comprising of heterogeneous or mixed cell populations is described. The killing or inhibition of tumors is achieved by selectively targeting a unique ligand suspected of being expressed on a particular cell population to also kill a cell population lacking the unique ligand. These conjugates have therapeutic use as they are delivered to a specific cell population to kill these cells and the cytotoxic drug is released to kill non-targeted cells, thereby eliminating the tumor.

Claims

exact text as granted — not AI-modified
1 . A method of eliminating a heterogeneous or mixed cell population AB, wherein A represents a cell population selectively expressing an ligand and B represents a cell lacking the ligand expressed in A, said method comprising: 
 a. targeting a cell-binding agent drug conjugate to the selected cell population A, wherein one or more drugs is covalently linked to the cell-binding agent via a linker comprising a disulfide bond and the cell-binding agent binds to the ligand in the cell population A;    b. internalization of the cell-binding agent drug conjugate in the cells comprising the cell population A;    c. cleaving the drug from the conjugate such that the drug undergoes an alkylation step prior to killing the cells in the cell population A; and    d. releasing the alkylated drug from the cell population A to induce killing of cells in the cell population B,    thereby eliminating the heterogeneous cell population.    
     
     
         2 . The method of  claim 1 , wherein the linker is cleaved through disulfide exchange, at acid pH, by light or by peptidases  
     
     
         3 . The method of  claim 1  or  2 , wherein the drug is at least one maytansinoid, taxane or a CC1065, or an analog thereof.  
     
     
         4 . The method of  claim 1  or  2 , wherein the linker bears an alkyl group on a carbon atom adjoining the disulfide bond within the linker.  
     
     
         5 . The method of  claim 4 , wherein the alkyl group on the drug side of the conjugate.  
     
     
         6 . The method of  claim 1 , wherein the cellular alkylation step results in methylation of the free drug.  
     
     
         7 . The method of  claim 1 , wherein the drug is at least one maytansinoid  
     
     
         8 . The method of  claim 7 , wherein the linker is at any one of the C-3 hydroxyl, C-14 hydroxymethyl, C-15 hydroxyl or C-20 desmethyl groups of the at least one maytansinoid.  
     
     
         9 . The method of  claim 7 , wherein the at least one maytansinoid is an N-methyl-alanine-containing ester of maytansinol.  
     
     
         10 . The method of  claim 7 , wherein the at least one maytansinoid is an N-methyl-cysteine-containing ester of maytansinol.  
     
     
         11 . The method of  claim 7 , wherein the at least one maytansinoid is represented by formula (II′-L), (II′-D) or (II′-D,L):  
       
         
           
           
               
               
           
         
       
       wherein:  
       Y 1 ′ represents  
       (CR 7 R 8 ) l (CR 9 ═CR 10 ) p (C═C) q A o (CR 5 R 6 ) m D u (CR 11 ═CR 12 ) r (C═C) s B t (CR 3 R 4 ) n CR 1 R 2 S—, wherein: 
 R 1  to R 12  are each independently linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition, R 2  to R 12  can be H;  
 A, B, and D, each independently is cyclic alkyl or cyclic alkenyl having 3 to 10 carbon atoms, simple or substituted aryl, or heterocyclic aromatic or heterocycloalkyl radical;  
 l, m, n, o, p, q, r, s, t and u are each independently 0 or an integer of 1 to 5, provided that at least two of l, m, n, o, p, q, r, s, t and u are not both zero; and  
 May represents a maytansinoid that bears a side chain at C-3 hydroxyl, C-14 hydroxymethyl, C-15 hydroxyl or C-20 desmethyl.  
 
     
     
         12 . The method of  claim 1 , wherein R 1  is methyl and R 2  is H or R 1  and R 2  are methyl.  
     
     
         13 . The method of  claim 11 , wherein R 1  is methyl, R 2  is H, R 5 , R 6 , R 7  and R 8  are each H, I and m are each 1, and n is 0; or wherein R 1  and R 2  are methyl, R 5 , R 6 , R 7 , R 8  are each H, land mare 1, and n is O.  
     
     
         14 . The method of  claim 7 , wherein the at least one maytansinoid is represented by formula (II-L), (II-D), or (II-D,L):  
       
         
           
           
               
               
           
         
       
       wherein:  
       Y 1  represents (CR 7 R 8 ) l (CR 5 R 6 ) m (CR 3 R 4 ) n CR 1 R 2 S—, wherein: 
 R 1  to R 8  are each independently, linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition R 2  to R 8  can be H;  
 l, m and n are each independently an integer of 1 to 5, and in addition n can be 0; and  
 May represents a maytansinoid that bears the side chain at C-3 hydroxyl, C-14 hydroxymethyl, C-15 hydroxyl or C-20 desmethyl.  
 
     
     
         15 . The method of  claim 14 , wherein R 1  is methyl and R 2  is H or R 1  and R 2  are methyl.  
     
     
         16 . The method of  claim 14 , wherein R 1  is methyl, R 2  is H, R 5 , R 6 , R 7  and R 8  are each H, l and m are each 1, and n is 0; or wherein R 1  and R 2  are methyl, R 5 , R 6 , R 7 , R 8  are each H, l and m are 1, and n is 0.  
     
     
         17 . The method of  claim 7 , wherein the at least one maytansinoid is represented by formula 41′:  
       
         
           
           
               
               
           
         
       
       4 1 ′ 
       wherein:  
       Y 1 ′ represents  
       (CR 7 R 8 ) l (CR 9 ═CR 10 ) p (C═C) q A o (CR 5 R 6 ) m D u (CR 11 ═CR 12 ) r (C═C) s B t (CR 3 R 4 ) n CR 1 R 2 S—,  
       wherein: 
 R 1  to R 12  are each independently linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition, R 2  to R 12  can be H;  
 A, B, and D, each independently is cyclic alkyl or cyclic alkenyl having 3 to 10 carbon atoms, simple or substituted aryl, or heterocyclic aromatic or heterocycloalkyl radical; and  
 l, m, n, o, p, q, r, s, t and u are each independently 0 or an integer of 1 to 5, provided that at least two of l, m, n, o, p, q, r, s, t and u are not both zero.  
 
     
     
         18 . The method of  claim 17 , wherein R 1  is methyl and R 2  is H or R 1  and R 2  are methyl.  
     
     
         19 . The method of  claim 17 , wherein R1 is methyl, R 2  is H, R 5 , R 6 , R 7  and R 8  are each H, 1 and m are each 1, and n is 0; or wherein R 1  and R 2  are methyl, R 5 , R 6 , R 7 , R 8  are each H, l and mare 1, and n is O.  
     
     
         20 . The method of  claim 7 , wherein the at least one maytansinoid is represented by formula 4 1 :  
       
         
           
           
               
               
           
         
       
       wherein:  
       Y 1  represents (CR 7 R 8 ) l (CR 5 R 6 ) m (CR 3 R 4 ) n CR 1 R 2 S—, wherein: 
 R 1  to R 8  are each independently, linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition R 2  to R 8  can be H; and  
 l, m and n are each independently an integer of 1 to 5, and in addition n can be 0.  
 
     
     
         21 . The method of  claim 20 , wherein R 1  is methyl and R 2  is H or R 1  and R 2  are methyl.  
     
     
         22 . The method of  claim 20 , wherein R 1  is methyl, R 2  is H, R 5 , R 6 , R 7  and R 8  are each H, 1 and m are each 1, and n is 0; or wherein R 1  and R 2  are methyl, R 5 , R 6 , R 7 , R 8  are each H, l and m are 1, and n is O.  
     
     
         23 . The method of  claim 7 , wherein the at least one maytansinoid is DM1.  
     
     
         24 . The method of  claim 7 , wherein the at least one maytansinoid is DM3.  
     
     
         25 . The method of  claim 7 , wherein the at least one maytansinoid is DM4.  
     
     
         26 . The method of  claim 1 , wherein the cell-binding agent binds to tumor cells; virus infected cells, microorganism infected cells, parasite infected cells, autoimmune cells, activated cells, myeloid cells, activated T-cells, B cells, or melanocytes; cells expressing the CD33, CD19, CanAg, CALLA, or Her-2 antigens; or cells expressing insulin growth factor receptor, epidermal growth factor receptor, or folate receptor.  
     
     
         27 . The method of  claim 1 , wherein the cell population B are tumor cells; virus infected cells, microorganism infected cells, parasite infected cells, autoimmune cells, activated cells, myeloid cells, activated T-cells, B cells, or melanocytes or combinations thereof.  
     
     
         28 . The method of  claim 1 , wherein the ligand expressed by targeted cell population A is an antigen selected for CD33, CD19, CanAg, CALLA, or Her-2 antigens and wherein the non-targeted cell population B is lacking in the antigen targeted in cell population A.  
     
     
         29 . the method of  claim 1 , wherein the targeted cell population A is suspected of expressing insulin growth factor receptor, epidermal growth factor receptor, or folate receptor and wherein the non-targeted cell population B is lacking in the receptor targeted in cell population A.  
     
     
         30 . The method of  claim 1 , wherein the cell-binding agent binds to cells selected from the group consisting of breast cancer cells, prostate cancer cells, ovarian cancer cells, colorectal cancer cells, gastric cancer cells, squamous cancer cells, small-cell lung cancer cells, testicular cancer cells, and neuroblastoma cells expressing a particular ligand.  
     
     
         31 . The method of  claim 1 , wherein the cell population B is selected from the group consisting of breast cancer cells, prostate cancer cells, ovarian cancer cells, colorectal cancer cells, gastric cancer cells, squamous cancer cells, small-cell lung cancer cells, testicular cancer cells, and neuroblastoma cells.  
     
     
         32 . The method of  claim 24 , wherein the tumor cell is from a heterogeneous tumor.  
     
     
         33 . The method of  claim 26 , wherein the heterogeneous tumor is a solid tumor.  
     
     
         34 . The method of  claim 1 , wherein the cell-binding agent is an antibody, a single chain antibody, an antibody fragment that specifically binds to a target cell, a monoclonal antibody, a single chain monoclonal antibody, or a monoclonal antibody fragment that specifically binds to a target cell, a chimeric antibody, a chimeric antibody fragment that specifically binds to a target cell, a domain antibody, a domain antibody fragment that specifically binds to a target cell, a lymphokine, a hormone, a vitamin, a growth factor, a colony stimulating factor, or a nutrient-transport molecule.  
     
     
         35 . The method of  claim 1 , wherein the cell-binding agent is an interferon, IL2, IL3, IL4, IL6, insulin, thyrotropin releasing hormone, melanocyte-stimulating hormone, a steroid hormone, somatostatin, EGF, TGF-α, FGF, G-CSF, VEGF, MCSF, GM-CSF, folic acid, transferrin, estrogen, estrogen analogues, androgen, or androgen analogues.  
     
     
         36 . The method of  claim 1 , wherein the cell-binding agent is an antibody, a single chain antibody, or an antibody fragment that specifically binds to a target cell.  
     
     
         37 . The method of  claim 1 , wherein the cell-binding agent is a resurfaced antibody, a resurfaced single chain antibody, or a resurfaced antibody fragment that specifically binds to a target cell.  
     
     
         38 . The method of  claim 1 , wherein the cell-binding agent is a humanized antibody, a humanized single chain antibody, or a humanized antibody fragment that specifically binds to a target cell.  
     
     
         39 . The method of  claim 1 , wherein the cell-binding agent is a monoclonal antibody, a single chain monoclonal antibody, or a monoclonal antibody fragment that specifically binds to a target cell.  
     
     
         40 . The method of  claim 1 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically binds to a target cell.  
     
     
         41 . The method of  claim 1 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically binds to a target cell.  
     
     
         42 . The method of  claim 1 , wherein the cell-binding agent is a chimeric antibody, a chimeric antibody fragment, a domain antibody, or a domain antibody fragment that specifically binds to a target cell.  
     
     
         43 . The method of  claim 1 , wherein the cell-binding agent is a monoclonal antibody, a single chain monoclonal antibody, or a monoclonal antibody fragment that specifically binds to tumor cells.  
     
     
         44 . The method of  claim 1 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically binds to tumor cells.  
     
     
         45 . The method of  claim 1 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically binds to tumor cells.  
     
     
         46 . The method of  claim 1 , wherein the cell-binding agent is a chimeric antibody, a chimeric antibody fragment, a domain antibody, or a domain antibody fragment that specifically binds to tumor cells.  
     
     
         47 . The method of  claim 1 , wherein the cell-binding agent is a monoclonal antibody, a single chain monoclonal antibody, or a monoclonal antibody fragment that specifically binds to colorectal cancer cells or breast cancer cells.  
     
     
         48 . The method of  claim 1 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically binds to colorectal cancer cells or breast cancer cells.  
     
     
         49 . The method of  claim 1 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically binds to colorectal cancer cells or breast cancer cells.  
     
     
         50 . The method of  claim 1 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically binds to breast cancer cells.  
     
     
         51 . The method of  claim 1 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically binds to breast cancer cells.  
     
     
         52 . The method of any one of claims  1 , wherein the cell-binding agent is an anti-CanAg antibody, an anti-CD19 antibody, an anti-CD33 antibody, an anti-CALLA antibody, an anti-EGFR antibody, an anti-CD56 antibody, an anti-IGF-R antibody, or an anti-Her2 antibody  
     
     
         53 . The method of  claim 1 , wherein the cell-binding agent is resurfaced antibody My9-6, KS77, or N901.  
     
     
         54 . The method of  claim 1 , wherein the cell-binding agent is the trastuzumab antibody, B4 antibody, or huC242 antibody.  
     
     
         55 . The method of  claim 1 , wherein the cell-binding agent is the huC242 antibody.  
     
     
         56 . The method of  claim 1 , wherein the cell-binding agent is the trastuzumab antibody.  
     
     
         57 . A method of eliminating cells in an abnormal cell population, said method comprising: 
 a) targeting a cell-binding agent drug conjugate to a selected cell population that expresses a ligand recognized by the cell-binding agent of the conjugate, wherein one or more drugs is covalently linked to the cell-binding agent via a linker comprising a disulfide bond and the cell-binding agent binds to the cells;    b) conversion of the pro-drug in the conjugate to a cytotoxic drug which further undergoes an alkylation step prior to killing the selected cell population suspected of containing the ligand; and    c) releasing the alkylated cytotoxic drug from the selected cell population to induce killing of another selected cell population suspected of lacking the ligand, thereby eliminating cells in the abnormal cell population.    
     
     
         58 . The method of  claim 57 , wherein the linker is cleaved through disulfide exchange, at acid pH, by light or by peptidases.  
     
     
         59 . The method of  claim 57  or  58 , wherein the drug is at least one maytansinoid, taxane or a CC1065, or an analog thereof.  
     
     
         60 . The method of  claim 57 , wherein the linker bears an alkyl group on a carbon atom adjoining the disulfide bond within the linker.  
     
     
         61 . The method of  claim 57 , wherein the alkyl group on the drug side of the conjugate.  
     
     
         62 . The method of  claim 57 , wherein the drug is at least one maytansinoid.  
     
     
         63 . The method of  claim 62 , wherein the linker is at any one of the C-3 hydroxyl, C-14 hydroxymethyl, C-15 hydroxyl or C-20 desmethyl groups of the at least one maytansinoid.  
     
     
         64 . The method of  claim 62 , wherein the at least one maytansinoid is an N-methyl-alanine-containing ester of maytansinol.  
     
     
         65 . The method of  claim 62 , wherein the at least one maytansinoid is an N-methyl-cysteine-containing ester of maytansinol.  
     
     
         66 . The method of  claim 62 , wherein the at least one maytansinoid is represented by formula (II′-L), (II′-D) or (II′-D,L):  
       
         
           
           
               
               
           
         
       
       wherein:  
       Y 1 ′ represents  
       (CR 7 R 8 ) l (CR 9 ═CR 10 ) p (C═C) q A o (CR 5 R 6 ) m D u (CR 11 ═CR 12 ) r (C═C) s B t (CR 3 R 4 ) n CR 1 R 2 S—,  
       wherein: 
 R 1  to R 12  are each independently linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition, R 2  to R 12  can be H;  
 A, B, and D, each independently is cyclic alkyl or cyclic alkenyl having 3 to 10 carbon atoms, simple or substituted aryl, or heterocyclic aromatic or heterocycloalkyl radical;  
 l, m, n, o, p, q, r, s, t and u are each independently 0 or an integer of 1 to 5, provided that at least two of l, m, n, o, p, q, r, s, t and u are not both zero; and  
 May represents a maytansinoid that bears a side chain at C-3 hydroxyl, C-14 hydroxymethyl, C-15 hydroxyl or C-20 desmethyl.  
 
     
     
         67 . The method of  claim 66 , wherein R 1  is methyl and R 2  is H or R 1  and R 2  are methyl.  
     
     
         68 . The method of  claim 66 , wherein R 1  is methyl, R 2  is H, R 5 , R 6 , R 7  and R 8  are each H, l and m are each 1, and n is 0; or wherein R 1  and R 2  are methyl, R 5 , R6, R 7 , R 8  are each H, l and m are 1, and n is 0.  
     
     
         69 . The method of  claim 62 , wherein the at least one maytansinoid is represented by formula (II-L), (II-D), or (II-D,L):  
       
         
           
           
               
               
           
         
       
       wherein:  
       Y 1  represents (CR 7 R 8 ) l (CR 5 R 6 ) m (CR 3 R 4 ) n CR 1 R 2 S—, wherein: 
 R 1  to R 8  are each independently, linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition R 2  to R 8  can be H;  
 l, m and n are each independently an integer of 1 to 5, and in addition n can be 0; and  
 May represents a maytansinoid that bears the side chain at C-3 hydroxyl, C-14 hydroxymethyl, C-15 hydroxyl or C-20 desmethyl.  
 
     
     
         70 . The method of  claim 69 , wherein R 1  is methyl and R 2  is H or R 1  and R 2  are methyl.  
     
     
         71 . The method of  claim 69 , wherein R 1  is methyl, R 2  is H, R 5 , R 6 , R 7  and R 8  are each H, l and m are each 1, and n is 0; or wherein R 1  and R 2  are methyl, R 5 , R 6 , R 7 , R 9  are each H, l and m are 1, and n is O.  
     
     
         72 . The method of  claim 62 , wherein the at least one maytansinoid is represented by formula 4 1 ′:  
       
         
           
           
               
               
           
         
       
       4 1 ′ 
       wherein: 
 Y 1 ′ represents  
 (CR 7 R 8 ) l (CR 9 ═CR 10 ) p (C═C) q A o (CR 5 R 6 ) m D u (CR 11 ═CR 12 ) r (C═C) s B t (CR 3 R 4 ) n CR 1 R 2 S—,  
 wherein:  
 R 1  to R 12  are each independently linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition, R 2  to R 12  can be H;  
 A, B, and D, each independently is cyclic alkyl or cyclic alkenyl having 3 to 10 carbon atoms, simple or substituted aryl, or heterocyclic aromatic or heterocycloalkyl radical; and  
 l, m, n, o, p, q, r, s, t and u are each independently 0 or an integer of 1 to 5, provided that at least two of l, m, n, o, p, q, r, s, t and u are not both zero.  
 
     
     
         73 . The method of  claim 72 , wherein R 1  is methyl and R 2  is H or R 1  and R 2  are methyl.  
     
     
         74 . The method of  claim 72 , wherein R 1  is methyl, R 2  is H, R 5 , R 6 , R 7  and R 8  are each H, 1 and m are each 1, and n is 0; or wherein R 1  and R 2  are methyl, R 5 , R 6 , R 7 , R 8  are each H, l and m are 1, and n is 0.  
     
     
         75 . The method of  claim 62 , wherein the at least one maytansinoid is represented by formula 4 1 :  
       
         
           
           
               
               
           
         
       
       wherein:  
       Y 1  represents (CR 7 R 8 ) l (CR 5 R 6 ) m (CR 3 R 4 ) n CR 1 R 2 S—, wherein: 
 R 1  to R 8  are each independently, linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition R 2  to R8 can be H; and  
 l, m and n are each independently an integer of 1 to 5, and in addition n can be 0.  
 
     
     
         76 . The method of  claim 75 , wherein R 1  is methyl and R 2  is H or R 1  and R 2  are methyl.  
     
     
         77 . The method of  claim 75 , wherein R 1  is methyl, R 2  is H, R 5 , R 6 , R 7  and R 8  are each H, 1 and m are each 1, and n is 0; or wherein R 1  and R 2  are methyl, R 5 , R 6 , R 7 , R 9  are each H, l and m are 1, and n is 0.  
     
     
         78 . The method of  claim 62 , wherein the at least one maytansinoid is DM 1.  
     
     
         79 . The method of  claim 62 , wherein the at least one maytansinoid is DM3.  
     
     
         80 . The method of  claim 62 , wherein the at least one maytansinoid is DM4.  
     
     
         81 . The method of  claim 57 , wherein the cell-binding agent binds to tumor cells; 
 virus infected cells, microorganism infected cells, parasite infected cells, autoimmune cells, activated cells, myeloid cells, activated T-cells, B cells, or melanocytes; cells expressing the CD33, CD19, CanAg, CALLA, or Her-2 antigens; or cells expressing insulin growth factor receptor, epidermal growth factor receptor, or folate receptor.    
     
     
         82 . The method of  claim 57 , wherein the cell-binding agent binds to cells selected from the group consisting of breast cancer cells, prostate cancer cells, ovarian cancer cells, colorectal cancer cells, gastric cancer cells, squamous cancer cells, small-cell lung cancer cells, testicular cancer cells, and neuroblastoma cells.  
     
     
         83 . The method of  claim 57 , wherein the cell-binding agent is an antibody, a single chain antibody, an antibody fragment that specifically binds to a target cell, a monoclonal antibody, a single chain monoclonal antibody, or a monoclonal antibody fragment that specifically binds to a target cell, a chimeric antibody, a chimeric antibody fragment that specifically binds to a target cell, a domain antibody, a domain antibody fragment that specifically binds to a target cell, a lymphokine, a hormone, a vitamin, a growth factor, a colony stimulating factor, or a nutrient-transport molecule.  
     
     
         84 . The method of  claim 57 , wherein the cell-binding agent is an interferon, IL2, IL3, IL4, IL6, insulin, thyrotropin releasing hormone, melanocyte-stimulating hormone, a steroid hormone, somatostatin, EGF, TGF-α, FGF, G-CSF, VEGF, MCSF, GM-CSF, folic acid, transferrin, estrogen, estrogen analogues, androgen, or androgen analogues.  
     
     
         85 . The method of  claim 57 , wherein the cell-binding agent is an antibody, a single chain antibody, or an antibody fragment that specifically binds to a target cell.  
     
     
         86 . The method of  claim 57 , wherein the cell-binding agent is a resurfaced antibody, a resurfaced single chain antibody, or a resurfaced antibody fragment that specifically binds to a target cell.  
     
     
         87 . The method of  claim 57 , wherein the cell-binding agent is a humanized antibody, a humanized single chain antibody, or a humanized antibody fragment that specifically binds to a target cell.  
     
     
         88 . The method of  claim 57 , wherein the cell-binding agent is a monoclonal antibody, a single chain monoclonal antibody, or a monoclonal antibody fragment that specifically binds to a target cell.  
     
     
         89 . The method of  claim 57 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically binds to a target cell.  
     
     
         90 . The method of  claim 57 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically binds to a target cell.  
     
     
         91 . The method of  claim 57 , wherein the cell-binding agent is a chimeric antibody, a chimeric antibody fragment, a domain antibody, or a domain antibody fragment that specifically binds to a target cell.  
     
     
         92 . The method of  claim 57 , wherein the cell-binding agent is a monoclonal antibody, a single chain monoclonal antibody, or a monoclonal antibody fragment that specifically binds to tumor cells.  
     
     
         93 . The method of  claim 57 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically binds to tumor cells.  
     
     
         94 . The method of  claim 57 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically binds to tumor cells.  
     
     
         95 . The method of  claim 57 , wherein the cell-binding agent is a chimeric antibody, a chimeric antibody fragment, a domain antibody, or a domain antibody fragment that specifically binds to tumor cells.  
     
     
         96 . The method of  claim 57 , wherein the cell-binding agent is a monoclonal antibody, a single chain monoclonal antibody, or a monoclonal antibody fragment that specifically binds to colorectal cancer cells or breast cancer cells.  
     
     
         97 . The method of  claim 57 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically binds to colorectal cancer cells or breast cancer cells.  
     
     
         98 . The method of  claim 57 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically binds to colorectal cancer cells or breast cancer cells.  
     
     
         99 . The method of  claim 57 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically binds to breast cancer cells.  
     
     
         100 . The method of  claim 57 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically binds to breast cancer cells.  
     
     
         101 . The method of any one of claims  57 , wherein the cell-binding agent is an anti-CanAg antibody, an anti-CD19 antibody, an anti-CD33 antibody, an anti-CALLA antibody, an anti-EGFR antibody, an anti-CD56 antibody, an anti-IGF-R antibody, or an anti-Her2 antibody.  
     
     
         102 . The method of any one of  claim 57 , wherein the cell-binding agent is resurfaced antibody My9-6, KS77, or N901.  
     
     
         103 . The method of any one of  claim 57 , wherein the cell-binding agent is the trastuzumab antibody, B4 antibody, or huC242 antibody.  
     
     
         104 . The method of any one of  claim 57 , wherein the cell-binding agent is the huC242 antibody.  
     
     
         105 . The method of any one of  claim 57 , wherein the cell-binding agent is the trastuzumab antibody.  
     
     
         106 . A method of utilizing a targeted cell population to inhibit or kill non-targeted cells in a heterogeneous or mixed cell population, said method comprising: 
 a) targeting a cell-binding agent drug conjugate to the targeted cell population suspected of containing an ligand recognized by the cell-binding agent of the conjugate, wherein one or more drugs is covalently linked to the cell-binding agent via a linker comprising a disulfide bond and the cell-binding agent binds to the cells;    b) conversion of the pro-drug in the conjugate to a cytotoxic drug in the targeted selected cell population to inhibit or kill the targeted cells; and    c) releasing the cytotoxic drug from the targeted cell population to inhibit or kill the non-targeted cells which lack the ligand in the targeted cell,    d) thereby inhibit or kill the heterogeneous or mixed cell population, targeted and non-targeted.    
     
     
         107 . A method of killing tumor cells in a tumor comprising a heterogeneous or mixed cell population, said method comprising: 
 a) selectively targeting a shed ligand, shed from a selected population of tumor cells suspected of expressing the ligand with a cell-binding agent drug conjugate wherein one or more drugs is covalently linked to the cell-binding agent via a linker comprising a disulfide bond and the cell-binding agent binds to the shed ligand.    b) conversion of the pro-drug in the conjugate to a cytotoxic drug which further undergoes an alkylation step prior to killing the selected cell population of tumor cells which shed the ligand; and    c) further induce killing of tumor cells which completely lack the ligand, to thereby kill the heterogeneous or mixed cells in the tumor.    
     
     
         108 . A compound of formula: 
 MayOCH(Me)N(CH 3n )COCH 2 CH 2 (CMe 2 ) n SMe    wherein, n=0 or 1, and May=maytansinoid.    
     
     
         109 . A compound of formula: 
 MayOCH(Me)N(CH 3n )COCH 2 CH 2 (CMe 2 ) n SMe    wherein, DM1Me: n=0; DM4Me: n=1    
     
     
         110 . An alkylated maytansinoid, wherein the maytansinoid has undergone alkylation at an unreacted thiol group.  
     
     
         111 . The alkylated maytansinoid of  claim 110 , wherein the alkyl is methyl, ethyl, propyl or butyl.  
     
     
         112 . Isolated lysine-N ε -SMCC-DM1.  
     
     
         113 . Isolated lysine-N ε -SPDB-DM4.  
     
     
         114 . Isolated S-cysteinyl-DM4.  
     
     
         115 . Isolated S-methyl-DM4.

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