Method of targeting specific cell populations using cell-binding agent maytansinoid conjugates linked via a non-cleavable linker, said conjugates, and methods of making said conjugates
The present invention discloses a method for targeting maytansinoids to a selected cell population, the method comprising contacting a cell population or tissue suspected of containing the selected cell population with a cell-binding agent maytansinoid conjugate, wherein one or more maytansinoids is covalently linked to the cell-binding agent via a non-cleavable linker and the cell-binding agent binds to cells of the selected cell population.
1. A method for targeting maytansinoids to a selected cell population in vivo, the method comprising contacting a cell population or tissue suspected of containing the selected cell population with a cell-binding agent maytansinoid conjugate, wherein one or more maytansinoids is covalently linked to the cell-binding agent via a non-cleavable linker and the cell-binding agent binds to cells of the selected cell population,
wherein the cell-binding agent is an antibody, a single chain antibody or an antibody fragment that specifically bind to a target cell, wherein at least a constant region of the antibody, single chain antibody or antibody fragment comprises a human antibody sequence, and, provided that the cell-binding agent is not an anti-erbB antibody.
2. The method of claim 1 , wherein the non-cleavable linker is substantially resistant to acid-induced cleavage, light-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, and disulfide bond cleavage.
3. The method of claim 1 , wherein the non-cleavable linker has a sulfur atom.
4. The method of claim 3 , wherein the non-cleavable linker is derived from a maleimido-based moiety.
5. The method of claim 4 , wherein the non-cleavable linker is derived from a maleimido-based moiety selected from the group consisting of N-succinimidyl 4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), N-succinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxy-(6-amidocaproate) (LC-SMCC), κ-maleimidoundecanoic acid N-succinimidyl ester (KMUA), γ-maleimidobutyric acid N-succinimidyl ester (GMBS), ε-maleimidcaproic acid N-hydroxysuccinimide ester (EMCS), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), N-(α-maleimidoacetoxy)-succinimide ester (AMAS), succinimidyl-6-(β-maleimidopropionamido)hexanoate (SMPH), N-succinimidyl 4-(p-maleimidophenyl)-butyrate (SMPB), and N-(p-maleimidophenyl)isocyanate (PMPI).
6. The method of claim 5 , wherein the non-cleavable linker is derived from SMCC.
7. The method of claim 3 , wherein the non-cleavable linker is derived from a haloacetyl-based moiety.
8. The method of claim 7 , wherein the non-cleavable linker is derived from a haloacetyl-based moiety selected from the group consisting of N-succinimidyl-4-(iodoacetyl)-aminobenzoate (SIAB), N-succinimidyl iodoacetate (SIA), N-succinimidyl bromoacetate (SBA), and N-succinimidyl 3-(bromoacetamido)propionate (SBAP).
9. The method of claim 8 , wherein the non-cleavable linker is derived from SIAB.
10. The method of claim 1 , 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.
11. The method of claim 1 , wherein the at least one maytansinoid is an N-methyl-alanine-containing ester of maytansinol.
12. The method of claim 1 , wherein the at least one maytansinoid is an N-methyl-cysteine-containing ester of maytansinol.
13. The method of claim 1 , 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 C 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.
14. The method of claim 13 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.
15. The method of claim 13 , 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.
16. The method of claim 1 , 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.
17. The method of claim 16 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.
18. The method of claim 16 , 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.
19. The method of claim 1 , wherein the at least one maytansinoid is represented by formula 41′:
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.
20. The method of claim 19 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.
21. The method of claim 19 , 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.
22. The method of claim 1 , wherein the at least one maytansinoid is represented by formula 41:
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.
23. The method of claim 22 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.
24. The method of claim 22 , 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.
25. The method of claim 1 , wherein the at least one maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1).
26. The method of claim 1 , wherein the at least one maytansinoid is N 2′ -deacetyl-N 2′ -(4-mercapto-1-oxopentyl)-maytansine (DM3).
27. The method of claim 1 , wherein the at least one maytansinoid is N 2′ -deacetyl-N 2′ -(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4).
28. 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 in graft rejection or graft vs. host disease, myeloid cells, activated T-cells, B cells, or melanocytes; cells expressing the CD33, CD19, CanAg, or CALLA antigens; or cells expressing insulin growth factor receptor or folate receptor.
29. The method of claim 1 , wherein the cell-binding agent binds to breast cancer cells, kidney cancer cells, lung cancer cells, prostate cancer cells, ovarian cancer cells, colorectal cancer cells, gastric cancer cells, squamous cancer cells, small-cell lung cancer cells, non-small-cell lung cancer cells, pancreatic cancer cells, testicular cancer cells, neuroblastoma cells, melanoma cells, cells from cancer of the lymphatic organs or a combination thereof.
30. The method of claim 1 , wherein the cell-binding agent is said resurfaced antibody, said resurfaced single chain antibody, or said resurfaced antibody fragment that specifically bind to a target cell.
31. 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.
32. 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 bind to a target cell.
33. 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 bind to a target cell.
34. 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 bind to tumor cells.
35. 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 bind to tumor cells.
36. 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 bind to colorectal cancer cells, breast cancer cells, kidney cancer cells, lung cancer cells, prostate cancer cells, ovarian cancer cells, gastric cancer cells, squamous cancer cells, small-cell lung cancer cells, non-small-cell lung cancer cells, pancreatic cancer cells, testicular cancer cells, neuroblastoma cells, melanoma cells, cells from cancer of the lymphatic organs or a combination thereof.
37. 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 bind to colorectal cancer cells, breast cancer cells, kidney cancer cells, lung cancer cells, prostate cancer cells, ovarian cancer cells, gastric cancer cells, squamous cancer cells, small-cell lung cancer cells, non-small-cell lung cancer cells, pancreatic cancer cells, testicular cancer cells, neuroblastoma cells, melanoma cells, cells from cancer of the lymphatic organs or a combination thereof.
38. 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 bind to breast cancer cells.
39. 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 bind to breast cancer cells.
40. The method of any one of claims 1 to 2 and 3 to 27 , wherein the cell-binding agent is an anti-PSMA antibody, an anti-CanAg antibody, an anti-CD19 antibody, an anti-CD33 antibody, an anti-CALLA antibody, an anti-CD56 antibody, or an anti-IGF-IR antibody.
41. The method of any one of claims 1 to 2 and 3 to 27 , wherein the cell-binding agent is a resurfaced or humanized My9-6 or N901 antibody or fragment thereof, wherein said My9-6 antibody comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody My9-6, and wherein said light chain comprises three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody My9-6; and wherein said resurfaced or humanized N901 antibody or fragment thereof comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody N901, and said light chain comprising three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody N901.
42. The method of any one of claims 1 to 2 and 3 to 27 , wherein the cell-binding agent is a resurfaced or humanized B4 antibody, or a resurfaced or humanized C242 antibody or fragment thereof, wherein said resurfaced or humanized C242 antibody comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody C242, and said light chain comprising three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody C242.
43. The method of any one of claims 1 to 2 and 3 to 27 , wherein the cell-binding agent is a resurfaced or humanized C242 antibody or fragment thereof, wherein said resurfaced or humanized C242 antibody comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody C242, and said light chain comprising three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody C242.
44. A method of eliminating cells in vivo, the method comprising contacting the cells in vivo with a cell-binding agent maytansinoid conjugate, wherein one or more maytansinoids is covalently linked to the cell-binding agent via a non-cleavable linker and the cell-binding agent binds to the cells,
wherein the cell-binding agent is an antibody, a single chain antibody or an antibody fragment that specifically binds to a target cell, wherein at least a constant region of the antibody, single chain antibody or antibody fragment comprises a human antibody sequence, and provided that the cell-binding agent is not an anti-erbB antibody.
45. A method of treatment of afflictions selected from the group consisting of tumors, autoimmune diseases, graft rejections, graft versus host disease, viral infections, and parasite infections, the method comprising administering to a subject in need of treatment an effective amount of a cell-binding agent maytansinoid conjugate, wherein one or more maytansinoids is covalently linked to the cell-binding agent via a non-cleavable linker and the cell-binding agent binds diseased or infected cells of the affliction,
wherein the cell-binding agent is an antibody, a single chain antibody or an antibody fragment that specifically binds to diseased or infected cells of the affliction, wherein at least a constant region of the antibody, single chain antibody or antibody fragment comprises a human antibody sequence, and provided that the cell-binding agent is not an anti-erbB antibody.
46. The method of claim 44 , wherein the non-cleavable linker is substantially resistant to acid-induced cleavage, light-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, and disulfide bond cleavage.
47. The method of claim 44 , wherein the non-cleavable linker has a sulfur atom.
48. The method of claim 47 , wherein the non-cleavable linker is derived from a maleimido-based moiety.
49. The method of claim 48 , wherein the non-cleavable linker is derived from a maleimido-based moiety selected from the group consisting of N-succinimidyl 4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), N-succinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxy-(6-amidocaproate) (LC-SMCC), κ-maleimidoundecanoic acid N-succinimidyl ester (KMUA), γ-maleimidobutyric acid N-succinimidyl ester (GMBS), ε-maleimidcaproic acid N-hydroxysuccinimide ester (EMCS), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), N-(α-maleimidoacetoxy)-succinimide ester (AMAS), succinimidyl-6-(β-maleimidopropionamido)hexanoate (SMPH), N-succinimidyl 4-(p-maleimidophenyl)-butyrate (SMPB), and N-(p-maleimidophenyl)isocyanate (PMPI).
50. The method of claim 49 , wherein the non-cleavable linker is derived from SMCC.
51. The method of claim 47 , wherein the non-cleavable linker is derived from a haloacetyl-based moiety.
52. The method of claim 51 , wherein the non-cleavable linker is derived from a haloacetyl-based moiety selected from the group consisting of N-succinimidyl-4-(iodoacetyl)-aminobenzoate (SIAB), N-succinimidyl iodoacetate (SIA), N-succinimidyl bromoacetate (SBA), and N-succinimidyl 3-(bromoacetamido)propionate (SBAP).
53. The method of claim 52 , wherein the non-cleavable linker is derived from SIAB.
54. The method of claim 44 , 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.
55. The method of claim 44 , wherein the at least one maytansinoid is an N-methyl-alanine-containing ester of maytansinol.
56. The method of claim 54 , wherein the at least one maytansinoid is an N-methyl-cysteine-containing ester of maytansinol.
57. The method of claim 44 , 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.
58. The method of claim 57 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.
59. The method of claim 57 , 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.
60. The method of claim 44 , 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.
61. The method of claim 60 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.
62. The method of claim 60 , 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.
63. The method of claim 44 , wherein the at least one maytansinoid is represented by formula 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.
64. The method of claim 63 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.
65. The method of claim 64 , 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.
66. The method of claim 44 , 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.
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 , R 6 , R 7 , R 8 are each H, l and m are 1, and n is 0.
69. The method of claim 44 , wherein the at least one maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1).
70. The method of claim 44 , wherein the at least one maytansinoid is N 2′ -deacetyl-N 2′ -(4-mercapto-1-oxopentyl)-maytansine (DM3).
71. The method of claim 44 , wherein the at least one maytansinoid is N 2′ -deacetyl-N 2′ -(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4).
72. The method of claim 44 , wherein the cell-binding agent binds to tumor cells; virus infected cells, microorganism infected cells, parasite infected cells, autoimmune cells, activated cells in graft rejection or graft vs. host disease, myeloid cells, activated T-cells, B cells, or melanocytes; cells expressing the CD33, CD19, CanAg, or CALLA antigens; or cells expressing insulin growth factor receptor or folate receptor.
73. The method of claim 44 , wherein the cell-binding agent binds to breast cancer cells, kidney cancer cells, lung cancer cells, prostate cancer cells, ovarian cancer cells, colorectal cancer cells, gastric cancer cells, squamous cancer cells, small-cell lung cancer cells, non-small-cell lung cancer cells, pancreatic cancer cells, testicular cancer cells, neuroblastoma cells, melanoma cells, cells from cancer of the lymphatic organs or a combination thereof.
74. The method of claim 44 , 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.
75. The method of claim 44 , 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.
76. The method of claim 44 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically bind to a target cell.
77. The method of claim 44 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically bind to a target cell.
78. The method of claim 44 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically bind to tumor cells.
79. The method of claim 44 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically bind to tumor cells.
80. The method of claim 44 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically bind to colorectal cancer cells or breast cancer cells.
81. The method of claim 44 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically bind to colorectal cancer cells or breast cancer cells.
82. The method of claim 44 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically bind to breast cancer cells.
83. The method of claim 44 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically bind to breast cancer cells.
84. The method of claim 44 , wherein the cell-binding agent is an anti-PSMA antibody, an anti-CanAg antibody, an anti-CD19 antibody, an anti-CD33 antibody, an anti-CALLA antibody, an anti-CD56 antibody, or an anti-IGF-IR antibody.
85. The method of claim 44 , wherein the cell-binding agent is a resurfaced or humanized My9-6 or N901 antibody or fragment thereof, wherein said My9-6 antibody comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody My9-6, and wherein said light chain comprises three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody My9-6; and wherein said resurfaced or humanized N901 antibody or fragment thereof comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody N901, and said light chain comprising three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody N901.
86. The method of claim 44 , wherein the cell-binding agent is a resurfaced or humanized B4 antibody, or a resurfaced or humanized C242 antibody or fragment thereof, wherein said resurfaced or humanized C242 antibody comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody C242, and said light chain comprising three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody C242.
87. The method of claim 44 , wherein the cell-binding agent is a humanized C242 antibody or fragment thereof, wherein said humanized C242 antibody comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody C242, and said light chain comprising three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody C242.
88. The method of claim 44 , wherein the cell-binding agent is a resurfaced C242 antibody or fragment thereof, wherein said resurfaced or humanized C242 antibody comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody C242, and said light chain comprising three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody C242.
89. The method of claim 45 , wherein the tumors are selected from the group consisting of lung cancer, breast cancer, colorectal cancer, gastric cancer, squamous cell cancer, small-cell lung cancer, non-small-cell lung cancer cells, pancreatic cancer cells, testicular cancer, neuroblastoma cells, prostate cancer, kidney cancer, cancer of the pancreas, cancer of the ovary and cancer of the lymphatic organs.
90. The method of claim 45 , wherein the non-cleavable linker is substantially resistant to acid-induced cleavage, light-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, and disulfide bond cleavage.
91. The method of claim 45 , wherein the non-cleavable linker has a sulfur atom.
92. The method of claim 91 , wherein the non-cleavable linker is derived from a maleimido-based moiety.
93. The method of claim 92 , wherein the non-cleavable linker is derived from a maleimido-based moiety selected from the group consisting of N-succinimidyl 4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), N-succinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxy-(6-amidocaproate) (LC-SMCC), κ-maleimidoundecanoic acid N-succinimidyl ester (KMUA), γ-maleimidobutyric acid N-succinimidyl ester (GMBS), E-maleimidcaproic acid N-hydroxysuccinimide ester (EMCS), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), N-(α-maleimidoacetoxy)-succinimide ester (AMAS), succinimidyl-6-(β-maleimidopropionamido)hexanoate (SMPH), N-succinimidyl 4-(p-maleimidophenyl)-butyrate (SMPB), and N-(p-maleimidophenyl)isocyanate (PMPI).
94. The method of claim 93 , wherein the non-cleavable linker is derived from SMCC.
95. The method of claim 91 , wherein the non-cleavable linker is derived from a haloacetyl-based moiety.
96. The method of claim 95 , wherein the non-cleavable linker is derived from a haloacetyl-based moiety selected from the group consisting of N-succinimidyl-4-(iodoacetyl)-aminobenzoate (SIAB), N-succinimidyl iodoacetate (SIA), N-succinimidyl bromoacetate (SBA), and N-succinimidyl 3-(bromoacetamido)propionate (SBAP).
97. The method of claim 96 , wherein the non-cleavable linker is derived from SIAB.
98. The method of claim 45 , 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.
99. The method of claim 45 , wherein the at least one maytansinoid is an N-methyl-alanine-containing ester of maytansinol.
100. The method of claim 45 , wherein the at least one maytansinoid is an N-methyl-cysteine-containing ester of maytansinol.
101. The method of claim 45 , 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.
102. The method of claim 101 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.
103. The method of claim 101 , 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.
104. The method of claim 45 , 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.
105. The method of claim 104 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.
106. The method of claim 104 , 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.
107. The method of claim 45 , wherein the at least one maytansinoid is represented by formula 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.
108. The method of claim 107 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.
109. The method of claim 107 , 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.
110. The method of claim 45 , 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.
111. The method of claim 110 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.
112. The method of claim 110 , 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.
113. The method of claim 45 , wherein the at least one maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1).
114. The method of claim 45 , wherein the at least one maytansinoid is N 2′ -deacetyl-N 2′ -(4-mercapto-1-oxopentyl)-maytansine (DM3).
115. The method of claim 45 , wherein the at least one maytansinoid is N 2′ -deacetyl-N 2′ -(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4).
116. The method of claim 45 , wherein the cell-binding agent is said resurfaced antibody, said resurfaced single chain antibody, or said resurfaced antibody fragment that specifically bind to diseased or infected cells of the affliction.
117. The method of claim 45 , wherein the cell-binding agent is said humanized antibody, said humanized single chain antibody, or said humanized antibody fragment that specifically bind to diseased or infected cells of the affliction.
118. The method of claim 45 , wherein the cell-binding agent is a resurfaced monoclonal antibody, a resurfaced single chain monoclonal antibody, or a resurfaced monoclonal antibody fragment that specifically bind to diseased or infected cells of the affliction.
119. The method of claim 45 , wherein the cell-binding agent is a humanized monoclonal antibody, a humanized single chain monoclonal antibody, or a humanized monoclonal antibody fragment that specifically bind to diseased or infected cells of the affliction.
120. The method of claim 45 , wherein the cell-binding agent is an anti-PSMA antibody, an anti-CanAg antibody, an anti-CD19 antibody, an anti-CD33 antibody, an anti-CALLA antibody, an anti-CD56 antibody, or an anti-IGF-IR antibody.
121. The method of claim 45 , wherein the cell-binding agent is a resurfaced or humanized My9-6 or N901 antibody or fragment thereof, wherein said My9-6 antibody comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody My9-6, and wherein said light chain comprises three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody My9-6; and wherein said resurfaced or humanized N901 antibody or fragment thereof comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody N901, and said light chain comprising three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody N901.
122. The method of claim 45 , wherein the cell-binding agent is a resurfaced or humanized B4 antibody, or a resurfaced or humanized C242 antibody or fragment thereof, wherein said resurfaced or humanized C242 antibody comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody C242, and said light chain comprising three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody C242.
123. The method of claim 45 , wherein the cell-binding agent is a humanized C242 antibody or fragment thereof, wherein said humanized C242 antibody comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody C242, and said light chain comprising three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody C242.
124. The method of claim 45 , wherein the cell-binding agent is a resurfaced C242 antibody or fragment thereof, wherein said resurfaced C242 antibody comprises a heavy chain and a light chain, said heavy chain comprising three complementarity determining regions comprising HCCDR1, HCCDR2 and HCCDR3 of murine antibody C242, and said light chain comprising three complementarity determining regions comprising LCCDR1, LCCDR2 and LCCDR3 of murine antibody C242.
125. The method of claim 1 , wherein an average of 1 to about 10 maytansinoids is covalently linked to the cell-binding agent.
126. The method of claim 1 , wherein the cell-binding agent is a fully human antibody, a fully human single chain antibody, or a fully human antibody fragment that specifically bind to a target cell.
127. The method of claim 1 , wherein the cell-binding agent is a fully human antibody, a fully human single chain antibody, or a fully human antibody fragment that specifically bind to tumor cells.
128. The method of claim 1 , wherein the cell-binding agent is a fully human antibody, a fully human single chain antibody, or a fully human antibody fragment, the maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1) and the linker is derived from SMCC.
129. 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 bind to target cells, the maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1) and the linker is derived from SMCC.
130. 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 bind to target cells, the maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1) and the linker is derived from SMCC.
131. The method of claim 44 , wherein an average of 1 to about 10 maytansinoids is covalently linked to the cell-binding agent.
132. The method of claim 44 , wherein the cell-binding agent is a fully human antibody, a fully human single chain antibody, or a fully human antibody fragment that specifically bind to a target cell.
133. The method of claim 44 , wherein the cell-binding agent is a fully human antibody, a fully human single chain antibody, or a fully human antibody fragment that specifically bind to tumor cells.
134. The method of claim 44 , wherein the cell-binding agent is a fully human antibody, a fully human single chain antibody, or a fully human antibody fragment, the maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1) and the linker is derived from SMCC.
135. The method of claim 44 , wherein the cell-binding agent is a resurfaced antibody, a resurfaced single chain antibody or a resurfaced antibody fragment that specifically bind to target cells, the maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1) and the linker is derived from SMCC.
136. The method of claim 44 , wherein the cell-binding agent is a humanized antibody, a humanized single chain antibody or a humanized antibody fragment that specifically bind to target cells, the maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1) and the linker is derived from SMCC.
137. The method of claim 45 , wherein an average of 1 to about 10 maytansinoids is covalently linked to the cell-binding agent.
138. The method of claim 45 , wherein the cell-binding agent is a fully human antibody, a fully human single chain antibody, or a fully human antibody fragment that specifically bind to diseased or infected cells of the affliction.
139. The method of claim 45 , wherein the cell-binding agent is a fully human antibody, a fully human single chain antibody, or a fully human antibody fragment, the maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1) and the linker is derived from SMCC.
140. The method of claim 45 , wherein the cell-binding agent is a resurfaced antibody, a resurfaced single chain antibody or a resurfaced antibody fragment that specifically bind to target cells, the maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1) and the linker is derived from SMCC.
141. The method of claim 45 , wherein the cell-binding agent is a humanized antibody, a humanized single chain antibody or a humanized antibody fragment that specifically bind to target cells, the maytansinoid is N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine (DM1) and the linker is derived from SMCC.
142. The method of claim 1 , wherein the cell-binding agent is a fully human antibody, a fully human single chain antibody, or a fully human antibody fragment that specifically bind to colorectal cancer cells, breast cancer cells, kidney cancer cells, lung cancer cells, prostate cancer cells, ovarian cancer cells, gastric cancer cells, squamous cancer cells, small-cell lung cancer cells, non-small-cell lung cancer cells, pancreatic cancer cells, testicular cancer cells, neuroblastoma cells, melanoma cells, cells from cancer of the lymphatic organs or a combination thereof.
143. The method of claim 1 , wherein the cell-binding agent is a fully human antibody, a fully human single chain antibody, or a fully human antibody fragment that specifically bind to colorectal cancer cells, breast cancer cells, kidney cancer cells, lung cancer cells, prostate cancer cells, ovarian cancer cells, gastric cancer cells, squamous cancer cells, small-cell lung cancer cells, non-small-cell lung cancer cells, pancreatic cancer cells, testicular cancer cells, neuroblastoma cells, melanoma cells, cells from cancer of the lymphatic organs or a combination thereof.