ANTI-TISSUE FACTOR ANTIBODY-DRUG CONJUGATES AND RELATED METHODS
Provided herein are antibodies that specifically bind to human tissue factor (TF), anti-TF antibody-drug conjugates (ADCs), and compositions comprising the antibodies or ADCs. Also provided herein are methods of making and using the antibodies or ADCs, such as therapeutic and diagnostic methods.
1 . An antibody-drug conjugate comprising:
a. an antigen binding protein (Ab) which binds to the extracellular domain of human Tissue Factor (TF) (SEQ ID NO:810), wherein the Ab comprises a VH-CDR1, a VH-CDR2, a VH-CDR3, a VL-CDR1, a VL-CDR2, and a VL-CDR3, wherein
i. the VH-CDR1 comprises SEQ ID NO: 872, the VH-CDR2 comprises SEQ ID NO: 873, the VH-CDR3 comprises SEQ ID NO: 874, the VL-CDR1 comprises SEQ ID NO: 875, the VL-CDR2 comprises SEQ ID NO: 876, and the VL-CDR3 comprises SEQ ID NO: 877,
ii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A3,
iii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A,
iv. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5,
v. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5-T, or
vi. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25G1;
and
b. one or more linker-toxin moieties represented by Formula IV:
wherein:
X is *—C(O)NHCH(CH 2 (R 2 ))— + , wherein * and + represent the respective points of attachment as indicated in Formula IV, or X is absent;
L is a linker;
! represents the point of attachment of L to the Ab, where L is attached to the Ab through a covalent bond;
R 1 is selected from the group consisting of:
wherein # and % represent the respective points of attachment as indicated in Formula IV; and
R 2 is phenyl.
2 . The antibody-drug conjugate of claim 1 , wherein R 1 is selected from the group consisting of:
3 . The antibody-drug conjugate of claim 1 or claim 2 , wherein X is absent.
4 . The antibody-drug conjugate of any one of claims 1 - 3 , wherein the linker-toxin moiety of Formula IV is represented by Formula V:
5 . The antibody-drug conjugate of claim 4 , wherein R 1 is selected from the group consisting of:
6 . The antibody-drug conjugate of claim 4 or claim 5 , wherein R 1 is selected from the group consisting of:
7 . The antibody-drug conjugate of any one of claims 4 - 6 , wherein R 1 is:
8 . The antibody-drug conjugate of any one of the preceding claims, wherein L is a cleavable linker.
9 . The antibody-drug conjugate of any one of the preceding claims, wherein L is a peptide-containing linker.
10 . The antibody-drug conjugate of any one the preceding claims, wherein L is a protease-cleavable linker.
11 . The antibody-drug conjugate of any one of claims 1 - 7 , wherein L is a linker selected from one of N-(β-maleimidopropyloxy)-N-hydroxy succinimide ester (BMPS), N-(ε-maleimidocaproyloxy) succinimide ester (EMCS), N-[γ-maleimidobutyryloxy]succinimide ester (GMBS), 1,6-hexane-bis-vinylsulfone (HBVS), succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxy-(6-amidocaproate) (LC-SMCC), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), 4-(4-N-Maleimidophenyl)butyric acid hydrazide (MPBH), succinimidyl 3-(bromoacetamido)propionate (SBAP), succinimidyl iodoacetate (SIA), succinimidyl (4-iodoacetyl)aminobenzoate (SIAB), N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP), N-succinimidyl-4-(2-pyridylthio)pentanoate (SPP), succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), succinimidyl 4-(p-maleimidophenyl)butyrate (SMPB), succinimidyl 6-[(β-maleimidopropionamido)hexanoate] (SMPH), iminothiolane (IT), sulfo-EMCS, sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC, sulfo-SMPB, and succinimidyl-(4-vinylsulfone)benzoate (SVSB).
12 . The antibody-drug conjugate of any one of claims 1 - 7 , wherein L comprises a poly(ethylene)glycol chain of the formula:
wherein g is an integer from 1-20.
13 . The antibody-drug conjugate of claim 12 , wherein g is 3.
14 . An antibody-drug conjugate of Formula VI:
wherein:
Ab represents a tissue factor (TF) antibody;
n is an integer greater than or equal to 1;
X is *—C(O)NHCH(CH 2 (R 2 ))— + , wherein * and + represent the respective points of attachment as indicated in Formula VI, or X is absent;
L is a linker;
R 1 is selected from the group consisting of:
wherein # and % represent the respective points of attachment as indicated in Formula VI; and
R 2 is phenyl; and wherein
the Ab comprises a VH-CDR1, a VH-CDR2, a VH-CDR3, a VL-CDR1, a VL-CDR2, and a VL-CDR3, wherein
i. the VH-CDR1 comprises SEQ ID NO: 872, the VH-CDR2 comprises SEQ ID NO: 873, the VH-CDR3 comprises SEQ ID NO: 874, the VL-CDR1 comprises SEQ ID NO: 875, the VL-CDR2 comprises SEQ ID NO: 876, and the VL-CDR3 comprises SEQ ID NO: 877,
ii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A3,
iii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A,
iv. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5,
v. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5-T, or
vi. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25G1.
15 . The antibody-drug conjugate of claim 14 , wherein R 1 is selected from the group consisting of:
16 . The antibody-drug conjugate of claim 14 or 15 , wherein X is absent.
17 . The antibody-drug conjugate of claim 15 or claim 16 , wherein R 1 is selected from the group consisting of:
18 . The antibody-drug conjugate of any one of claims 15 - 17 , wherein R 1 is:
19 . The antibody-drug conjugate of any one of claims 16 - 17 , wherein L is a cleavable linker.
20 . The antibody-drug conjugate of any one of claims 14 - 19 , wherein L is a peptide-containing linker.
21 . The antibody-drug conjugate of any one of claims 14 - 19 , wherein L is a protease-cleavable linker.
22 . The antibody-drug conjugate of any one of claims 14 - 19 , wherein L is a linker selected from one of N-(β-maleimidopropyloxy)-N-hydroxy succinimide ester (BMPS), N-(ε-maleimidocaproyloxy) succinimide ester (EMCS), N-[γ-maleimidobutyryloxy]succinimide ester (GMBS), 1,6-hexane-bis-vinylsulfone (HBVS), succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxy-(6-amidocaproate) (LC-SMCC), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), 4-(4-N-Maleimidophenyl)butyric acid hydrazide (MPBH), succinimidyl 3-(bromoacetamido)propionate (SBAP), succinimidyl iodoacetate (SIA), succinimidyl (4-iodoacetyl)aminobenzoate (SIAB), N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP), N-succinimidyl-4-(2-pyridylthio)pentanoate (SPP), succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), succinimidyl 4-(p-maleimidophenyl)butyrate (SMPB), succinimidyl 6-[(β-maleimidopropionamido)hexanoate] (SMPH), iminothiolane (IT), sulfo-EMCS, sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC, sulfo-SMPB, and succinimidyl-(4-vinylsulfone)benzoate (SVSB).
23 . The antibody-drug conjugate of any one of claims 14 - 19 , wherein L comprises a poly(ethylene)glycol chain of the formula:
wherein g is an integer from 1-20.
24 . The antibody-drug conjugate of claim 23 , wherein g is 3.
25 . The antibody-drug conjugate of claim 14 , wherein L is represented by Formula VII:
wherein:
Z represents a functional group that binds to a target group of the TF antibody;
D represents the point of attachment to the amino group as indicated in Formula VI;
Str is a stretcher;
AA 1 and AA 2 are each independently an amino acid, wherein AA 1 -[AA 2 ] m forms a protease cleavage site;
X 1 is a self-immolative group;
s is an integer selected from 0 and 1;
m is an integer selected from the group consisting of 1, 2, 3, and 4;
o is an integer selected from 0, 1, and 2.
26 . The antibody-drug conjugate of claim 25 , wherein n is an integer selected from the group consisting of 1, 2, 3, 4, and 5.
27 . The antibody-drug conjugate of claim 25 or claim 26 , wherein [Str] s is selected from the group consisting of alkylene, stretchers based on aliphatic acids, stretchers based on aliphatic diacids, stretchers based on aliphatic amines and stretchers based on aliphatic diamines.
28 . The antibody-drug conjugate of any one of claims 25 - 27 , wherein [Str] s is selected from the group consisting of diglycolate-based stretchers, malonate-based stretchers, caproate-based stretchers and caproamide-based stretchers.
29 . The antibody-drug conjugate of claim 25 or claim 26 , wherein [Str] s is selected from the group consisting of glycine-based stretchers, polyethylene glycol-based stretchers, and monomethoxy polyethylene glycol-based stretchers.
30 . The antibody-drug conjugate of claim 25 or claim 26 , wherein [Str] s is:
wherein
h is an integer from 1-20,
CC refers to the point of attachment to AA 1 ; and
DD refers to the point of attachment to Z.
31 . The antibody-drug conjugate of claim 25 or claim 26 , wherein [Str] s is selected from:
wherein:
EE and FF represent the points of attachment to Z and AA 1 , respectively;
R is selected from hydrogen and C 1 -C 6 alkyl;
each occurrence of p is independently an integer from 2 to 10; and
each occurrence of q is independently an integer from 1 to 10.
32 . The antibody-drug conjugate of claim 25 , claim 26 , or claim 31 , wherein [Str] s is selected from the group consisting of:
wherein:
EE and FF represent the points of attachment to Z and AA 1 , respectively;
each occurrence of p is independently an integer from 2 to 10; and
each occurrence of q is independently an integer from 1 to 10.
33 . The antibody-drug conjugate of claim 25 , claim 26 , claim 31 , or claim 32 , wherein [Str] s is selected from:
wherein:
EE and FF represent the points of attachment to Z and AA 1 , respectively;
each occurrence of p is independently an integer from 2 to 6, and q is an integer from 2 to 8.
34 . The antibody-drug conjugate of any one of claims 25 - 33 , wherein AA 1 -[AA 2 ] m is selected from Val-Lys, Ala-Lys, Phe-Lys, Val-Cit, Phe-Cit, Leu-Cit, Ile-Cit, Trp-Cit, Phe-Arg, Ala-Phe, Val-Ala, Met-Lys, Asn-Lys, Ile-Pro, Ile-Val, Asp-Val, His-Val, Met-(D)Lys, Asn-(D)Lys, Val-(D)Asp, NorVal-(D)Asp, Ala-(D)Asp, Me 3 Lys-Pro, PhenylGly-(D)Lys, Met-(D)Lys, Asn-(D)Lys, Pro-(D)Lys, Met-(D)Lys, Met-Cit-Val, Gly-Cit-Val, (D)Phe-Phe-Lys, (D)Ala-Phe-Lys, Gly-Phe-Leu-Gly, and Ala-Leu-Ala-Leu.
35 . The antibody-drug conjugate of any one of claims 25 - 34 , wherein m is selected from 1, 2 and 3.
36 . The antibody-drug conjugate of any one of claims 25 - 35 , wherein m is 1.
37 . The antibody-drug conjugate of any one of claims 25 - 36 , wherein AA 1 -[AA 2 ] m is a dipeptide selected from Val-Lys, Ala-Lys, Phe-Lys, Val-Cit, Phe-Cit, Leu-Cit, Ile-Cit and Trp-Cit.
38 . The antibody-drug conjugate of any one of claims 25 - 37 , wherein each X 1 is independently selected from p-aminobenzyloxycarbonyl (PABC), p-aminobenzyl ether (PABE) and methylated ethylene diamine (MED).
39 . The antibody-drug conjugate of claim 30 , wherein s is 1 and h is 3.
40 . The antibody-drug conjugate of any one of claims 25 - 39 , wherein s is 1.
41 . The antibody-drug conjugate of any one of claims 25 - 40 , wherein o is 0.
42 . An antibody drug-conjugate comprising a linker-toxin moiety of the Formula VIII:
wherein ## represents the point of attachment of the linker-toxin moiety to the TF antibody and the linker-toxin moiety is attached to the TF antibody through a covalent bond.
43 . An antibody-drug conjugate of Formula IX:
wherein:
Ab is a tissue factor (TF) antibody, wherein the Ab comprises a VH-CDR1, a VH-CDR2, a VH-CDR3, a VL-CDR1, a VL-CDR2, and a VL-CDR3, wherein
i. the VH-CDR1 comprises SEQ ID NO: 872, the VH-CDR2 comprises SEQ ID NO: 873, the VH-CDR3 comprises SEQ ID NO: 874, the VL-CDR1 comprises SEQ ID NO: 875, the VL-CDR2 comprises SEQ ID NO: 876, and the VL-CDR3 comprises SEQ ID NO: 877,
ii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A3,
iii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A,
iv. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5,
v. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5-T, or
vi. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25G1,
n is an integer greater than or equal to 1, and
the succinimidyl group is attached to the Ab through a covalent bond.
44 . The antibody-drug conjugate of claim 43 , wherein n is selected from the group consisting of 1, 2, 3, 4, and 5.
45 . The antibody-drug conjugate of claim 43 or 44 , wherein n is selected from the group consisting of 2, 3, and 4.
46 . An antibody-drug conjugate comprising a linker as represented by Formula X:
wherein:
## is the point of attachment to the antibody and the succinimidyl group is attached to the antibody through a covalent bond;
Y is one or more additional linker components, or is absent; and
D 1 is the point of attachment to a cytotoxic agent, and wherein
the Ab comprises a VH-CDR1, a VH-CDR2, a VH-CDR3, a VL-CDR1, a VL-CDR2, and a VL-CDR3, wherein
i. the VH-CDR1 comprises SEQ ID NO: 872, the VH-CDR2 comprises SEQ ID NO: 873, the VH-CDR3 comprises SEQ ID NO: 874, the VL-CDR1 comprises SEQ ID NO: 875, the VL-CDR2 comprises SEQ ID NO: 876, and the VL-CDR3 comprises SEQ ID NO: 877,
ii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A3,
iii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A,
iv. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5,
v. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5-T, or
vi. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25G1.
47 . An antibody-drug conjugate comprising a linker as represented by Formula XI:
wherein:
## is the point of attachment to the antibody and the succinimidyl group is attached to the antibody through a covalent bond;
Y is one or more additional linker components, or is absent; and
D 1 is the point of attachment to a cytotoxic agent, and wherein
the Ab comprises a VH-CDR1, a VH-CDR2, a VH-CDR3, a VL-CDR1, a VL-CDR2, and a VL-CDR3, wherein
i. the VH-CDR1 comprises SEQ ID NO: 872, the VH-CDR2 comprises SEQ ID NO: 873, the VH-CDR3 comprises SEQ ID NO: 874, the VL-CDR1 comprises SEQ ID NO: 875, the VL-CDR2 comprises SEQ ID NO: 876, and the VL-CDR3 comprises SEQ ID NO: 877,
ii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A3,
iii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A,
iv. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5,
v. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5-T, or
vi. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25G1.
48 . The antibody-drug conjugate of claim 46 or claim 47 , wherein the cytotoxic agent is selected from the group consisting of a diagnostic agent, a metal chelator, an enzyme, a fluorescent compound, a bioluminescent compound, or a chemiluminescent compound.
49 . The antibody-drug conjugate of claim 46 or claim 47 , wherein the cytotoxic agent is a cytotoxic payload having an improved safety profile.
50 . The antibody-drug conjugate of any one of the preceding claims, wherein the Ab comprises:
a. a VH sequence that is SEQ ID NO: 868 and a VL sequence that is SEQ ID NO: 869,
b. a VH that is SEQ ID NO: 151 and a VL sequence that is SEQ ID NO: 152,
c. a VH sequence that is SEQ ID NO: 113 and a VL sequence that is SEQ ID NO: 114,
d. a VH sequence that is SEQ ID NO: 189 and a VL sequence that is SEQ ID NO: 190,
e. a VH sequence that is SEQ ID NO: 836 and a VL sequence that is SEQ ID NO: 837, or
f. a VH sequence that is SEQ ID NO: 265 and a VL sequence that is SEQ ID NO: 266.
51 . The antibody-drug conjugate of any one of the preceding claims, wherein the Ab comprises:
a. a heavy chain sequence that is
QVQLVQSGAEVKKPGASVKVSCKASGYTFDx[V/A]YGISWVRQAPGQ
GLEWMGWIAPYx[N/S]GNTNYAQKLQGRVTMTTDTSTSTAYMELRSL
RSDDTAVYYCARDAGTYSPFGYGMDVWGQGTTVTVSSASTKGPSVF
PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL
QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDK
THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDP
EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG
KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVS
LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG
and a light chain sequence that is
DIQMTQSPSTLSASVGDRVTITCx[R/Q]ASx[Q/E]SIx[S/N]x[S/
N]WLAWYQQKPGKAPKLLIYKAx[S/Y]x[S/N]LEx[S/Y]GVPSRFS
GSGSGTEFTLTISSLQPDDFATYYCQx[Q/L]FQx[S/K]LPPFTFGGG
TKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV
DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ
GLSSPVTKSFNRGEC,
b. a heavy chain sequence that is
QVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWVRQAPGQGLE
WMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTA
VYYCARDAGTYSPFGYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSK
STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS
NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG
and a light chain sequence that is
DIQMTQSPSTLSASVGDRVTITCQASQSINNWLAWYQQKPGKAPKLLI
YKAYNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQLFQSLPPFT
FGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV
QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYA
CEVTHQGLSSPVTKSFNRGEC,
c. a heavy chain sequence that is
QVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWVRQAPGQGLE
WMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTA
VYYCARDAGTYSPFGYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSK
STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS
NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGOPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWOOG
NVFSCSVMHEALHNHYTQKSLSLSPG
and a light chain sequence that is
DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIY
KASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQFQSLPPFTF
GGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQ
WKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC
EVTHQGLSSPVTKSFNRGEC,
d. a heavy chain sequence that is
QVQLVQSGAEVKKPGASVKVSCKASGYTFDAYGISWVRQAPGQGLE
WMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTA
VYYCARDAGTYSPFGYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSK
STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS
NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG
and a light chain sequence that is
DIQMTQSPSTLSASVGDRVTITCRASESISNWLAWYQQKPGKAPKLLIY
KAYSLEYGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQFQKLPPFTF
GGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQ
WKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC
EVTHQGLSSPVTKSFNRGEC,
e. a heavy chain sequence that is
QVQLVQSGAEVKKPGASVKVSCKASGYTFRSYGISWVRQAPGQGLE
WMGWVAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDT
AVYYCARDAGTYSPYGYGMDVWGQGTTVTVSSASTKGPSVFPLAPSS
KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPC
PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKG
FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG
and a light chain sequence that is
DIQMTQSPSTLSASVGDRVTITCRASHSIDSWLAWYQQKPGKAPKLLI
YKASYLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQLFQSLPPFT
FGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV
QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYA
CEVTHQGLSSPVTKSFNRGEC,
or
f. a heavy chain sequence that is
QVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWVRQAPGQGLE
WMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTA
VYYCARDAGTYSPFGYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSK
STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS
NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG
and a light chain sequence that is
DIQMTQSPSTLSASVGDRVTITCRASESISNWLAWYQQKPGKAPKLLIY
KAYSLEYGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQFQKLPPFTF
GGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQ
WKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC
EVTHQGLSSPVTKSFNRGEC.
52 . An antibody-drug conjugate of Formula IX:
wherein:
Ab is a tissue factor (TF) antibody, wherein the Ab comprises a VH-CDR1, a VH-CDR2, a VH-CDR3, a VL-CDR1, a VL-CDR2, and a VL-CDR3 from the antibody designated 25A3, and
n is an integer greater than or equal to 1.
53 . The antibody-drug conjugate of claim 52 , wherein n is selected from the group consisting of 1, 2, 3, 4, and 5.
54 . The antibody-drug conjugate of claim 52 , wherein n is selected from the group consisting of 2, 3, and 4.
55 . The antibody-drug conjugate of any one of claims 52 - 54 , wherein the Ab comprises a VH sequence that is SEQ ID NO: 151 and a VL sequence that is SEQ ID NO: 152.
56 . The antibody-drug conjugate of any one of claims 52 - 55 , wherein the Ab comprises a full heavy chain sequence that is
QVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWVRQAPGQGLEWMGW
IAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDA
GTYSPFGYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGC
LVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG
TQTYICNVNHKPSNTKVDKRVEPKS C DKTHT C PP C PAPELLGGPSVFLFP
PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE
QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR
EPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS
PG
and a light chain sequence that is
DIQMTQSPSTLSASVGDRVTITCQASQSINNWLAWYQQKPGKAPKLLIYK
AYNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQLFQSLPPFTFG
GGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK
VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ
GLSSPVTKSFNRGE C .
57 . An antibody-drug conjugate of Formula IX:
wherein:
Ab is a tissue factor (TF) antibody, wherein the Ab comprises a heavy chain sequence that is
QVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWVRQAPGQGLEWMGWI
APYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDAGT
YSPFGYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK
DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY
ICNVNHKPSNTKVDKRVEPKS C DKTHT C PP C PAPELLGGPSVFLFPPKPKD
TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL
PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDG
SFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG
and a light chain sequence that is
DIQMTQSPSTLSASVGDRVTITCQASQSINNWLAWYQQKPGKAPKLLIYK
AYNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQLFQSLPPFTFG
GGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK
VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ
GLSSPVTKSFNRGEC,
and
n is an integer greater than or equal to 1.
58 . The antibody-drug conjugate of claim 57 , wherein n is selected from the group consisting of 1, 2, 3, 4, and 5.
59 . The antibody-drug conjugate of claim 57 , wherein n is selected from the group consisting of 2, 3, and 4.
60 . An antibody-drug conjugate comprising an antibody (Ab) and one or more linker-toxins of the following structure:
wherein:
Ab is a tissue factor (TF) antibody, wherein the Ab comprises a VH-CDR1, a VH-CDR2, a VH-CDR3, a VL-CDR1, a VL-CDR2, and a VL-CDR3 from the antibody designated 25A3;
the one or more linker-toxins are attached to the Ab through a covalent bond; and
## represents a point of attachment of the linker-toxin to the Ab.
61 . An antibody-drug conjugate composition comprising the antibody-drug conjugate of claim 60 , wherein the composition comprises a multiplicity of drug-antibody ratio (DAR) species, wherein the average DAR of the composition is 2-4.
62 . An antibody-drug conjugate comprising an antibody (Ab) and one or more linker-toxins of the following structure:
wherein:
Ab is a tissue factor (TF) antibody, wherein the Ab comprises a heavy chain sequence that is
QVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWVRQAPGQGLEWMGW
IAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDA
GTYSPFGYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGC
LVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG
TQTYICNVNHKPSNTKVDKRVEPKS C DKTHT C PP C PAPELLGGPSVFLFP
PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE
QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR
EPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS
PG
and a light chain sequence that is
DIQMTQSPSTLSASVGDRVTITCQASQSINNWLAWYQQKPGKAPKLLIYK
AYNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQLFQSLPPFTFG
GGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK
VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ
GLSSPVTKSFNRGEC,
and
the one or more linker-toxins are attached to the Ab through a covalent bond; and
## represents a point of attachment of the linker-toxin to the Ab.
63 . An antibody-drug conjugate composition comprising the antibody-drug conjugate of claim 62 , wherein the composition comprises a multiplicity of drug-antibody ratio (DAR) species, wherein the average DAR of the composition is 2-4.
64 . The antibody-drug conjugate of any one of the preceding claims, wherein the Ab is multispecific.
65 . The antibody-drug conjugate of any one of the preceding claims, wherein the Ab is a Fab, Fab′, F(ab′) 2 , Fv, scFv, (scFv) 2 , single chain antibody molecule, dual variable domain antibody, single variable domain antibody, linear antibody, or V domain antibody.
66 . The antibody-drug conjugate of any one of the preceding claims, wherein the antibody comprises a scaffold, optionally wherein the scaffold is Fc, optionally human Fc.
67 . The antibody-drug conjugate of any one of the preceding claims, wherein the antibody comprises a heavy chain constant region of a class selected from IgG, IgA, IgD, IgE, and IgM.
68 . The antibody-drug conjugate of claim 67 , wherein the antibody comprises a heavy chain constant region of the class IgG, wherein the heavy chain constant region is from a subclass selected from IgG1, IgG2, IgG3, and IgG4.
69 . The antibody-drug conjugate of claim 68 , wherein the antibody comprises a heavy chain constant region of IgG1.
70 . The antibody-drug conjugate of claim 66 , wherein the Fc comprises one or more modifications, wherein the one or more modifications result in increased half-life, increased antibody-dependent cellular cytotoxicity (ADCC), increased antibody-dependent cellular phagocytosis (ADCP), increased complement-dependent cytotoxicity (CDC), or decreased effector function, compared with the Fc without the one or more modifications.
71 . The antibody-drug conjugate of any one of claims 1 - 70 , wherein, upon administration to a tumor-bearing subject, the antibody-drug conjugate reduces tumor volume or inhibits tumor growth.
72 . The antibody-drug conjugate of any one of claims 1 - 71 , wherein, upon administration to a tumor-bearing subject, the antibody-drug conjugate reduces tumor volume by at least about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99%.
73 . The antibody-drug conjugate of any one of claims 1 - 72 , wherein, upon administration to a tumor-bearing subject, the antibody-drug conjugate inhibits tumor growth by at least about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99%.
74 . The antibody-drug conjugate of any one of claims 1 - 73 , wherein, upon administration of the antibody-drug conjugate to a subject, the antibody-drug conjugate does not result in measurable skin toxicity.
75 . The antibody-drug conjugate of any one of claims 1 - 73 , wherein, upon administration of the antibody-drug conjugate to a subject, the antibody-drug conjugate results in reduced skin toxicity relative to a different anti-TF ADC.
76 . The antibody-drug conjugate of any one of claims 1 - 75 , wherein administration of the antibody-drug conjugate to a subject does not require administration of one or more anti-inflammatory agents.
77 . The antibody-drug conjugate of any one of claims 1 - 75 , wherein administration of the antibody-drug conjugate to a subject results in a reduced need for one or more anti-inflammatory agents relative to a different anti-TF ADC.
78 . The antibody-drug conjugate of claim 76 or 77 , wherein the one or more anti-inflammatory agents comprises at least one of topical steroids and systemic steroids.
79 . The antibody-drug conjugate of any one of claims 1 - 78 , wherein, upon administration of the antibody-drug conjugate to a subject, the antibody-drug conjugate results in reduced or absent neutropenia in the subject relative to baseline levels.
80 . The antibody-drug conjugate of any one of claims 1 - 78 , wherein, upon administration of the antibody-drug conjugate to a subject, the antibody-drug conjugate does not alter, increase or decrease the number of monocytes in the subject relative to baseline levels.
81 . The antibody-drug conjugate of any one of claims 75 - 80 , wherein the different anti-TF ADC is identical to the antibody-drug conjugate except for being conjugated to MMAE.
82 . A pharmaceutical composition comprising the antibody-drug conjugate of any one of the preceding claims and a pharmaceutically acceptable carrier.
83 . A method of treating or preventing a disease or condition in a subject in need thereof, comprising administering to the subject an effective amount of the antibody-drug conjugate of any one of claims 1 - 81 or the pharmaceutical composition of claim 82 .
84 . A method of treating or delaying the onset of cancer in a subject in need thereof, comprising administering to the subject an effective amount of the antibody-drug conjugate of any one of claims 1 - 81 .
85 . The method of claim 83 , wherein the disease or condition is cancer.
86 . The method of claim 84 or 85 , wherein the cancer is selected from the group consisting of: head and neck cancer, ovarian cancer, gastric cancer, esophageal cancer, cervical cancer, prostate cancer, pancreatic cancer, estrogen receptors negative (ER−) breast cancer, progesterone receptors negative (PR−) breast cancer, HER2 negative (HER2−) triple negative breast cancer, glioblastoma, lung cancer, bladder cancer, melanoma, and kidney cancer.
87 . The method of claim 83 , wherein the disease or condition involves neovascularization.
88 . The method of claim 87 , wherein the disease or condition involving neovascularization is cancer.
89 . The method of claim 83 , wherein the disease or condition involves vascular inflammation.
90 . The method of any one of claims 83 - 89 , further comprising administering one or more additional therapeutic agents to the subject.
91 . The method of claim 90 , wherein the composition further comprises the one or more additional therapeutic agents.
92 . The method of claim 90 , wherein the additional therapeutic agent is formulated in a different pharmaceutical composition.
93 . The method of claim 90 , wherein the additional therapeutic agent is administered prior to administering the composition.
94 . The method of claim 90 , wherein the additional therapeutic agent is administered after administering the composition.
95 . The method of claim 90 , wherein the additional therapeutic agent is administered contemporaneously with the composition.
96 . The method of any one of the preceding claims, wherein the subject is a human subject.
97 . A method of killing a cancer cell, comprising contacting the cancer cell with an effective amount of the antibody-drug conjugate of any one of claims 1 - 81 .
98 . A process for preparing an antibody-drug conjugate, the process comprising:
(A) reacting a nucleophilic or an electrophilic group on an antigen binding protein (Ab) which binds to the extracellular domain of human Tissue Factor (TF) (SEQ ID NO:810) with a bifunctional linker to form an Ab-linker intermediate, and reacting the Ab-linker intermediate with the −NH 2 group of a compound of general Formula I:
wherein:
X is *—C(O)NHCH(CH 2 (R 2 ))— + , wherein * and + represent the respective points of attachment as indicated in Formula I, or X is absent;
R 1 is selected from the group consisting of:
wherein # and % represent the respective points of attachment as indicated in Formula I; and R 2 is phenyl,
to provide the antibody drug conjugate; or
(B) reacting the —NH 2 group on the compound of general Formula I with a bifunctional linker to form a linker-toxin intermediate, and reacting the linker-toxin intermediate with a nucleophilic or an electrophilic group on an antigen binding protein (Ab) which binds to the extracellular domain of human Tissue Factor (TF) (SEQ ID NO: 810) to provide the antibody-drug conjugate, wherein, in (A) or (B),
(a) the Ab comprises a VH-CDR1, a VH-CDR2, a VH-CDR3, a VL-CDR1, a VL-CDR2, and a VL-CDR3, wherein
i. the VH-CDR1 comprises SEQ ID NO: 872, the VH-CDR2 comprises SEQ ID NO: 873, the VH-CDR3 comprises SEQ ID NO: 874, the VL-CDR1 comprises SEQ ID NO: 875, the VL-CDR2 comprises SEQ ID NO: 876, and the VL-CDR3 comprises SEQ ID NO: 877,
ii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A3,
iii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A,
iv. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5,
v. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5-T, or
vi. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25G1; and
(b) the antibody-drug conjugate comprises one or more moieties represented by Formula IV:
wherein:
X is *—C(O)NHCH(CH 2 (R 2 ))— + , wherein * and + represent the respective points of attachment as indicated in Formula IV, or X is absent;
L is a linker;
! represents the point of attachment of L to the Ab, where L is attached to the Ab through a covalent bond;
R 1 is selected from the group consisting of:
wherein # and % represent the respective points of attachment as indicated in Formula VI; and
R 2 is phenyl.
99 . A process for preparing an antibody-drug conjugate, the process comprising:
(A) reacting a nucleophilic or an electrophilic group on an antigen binding protein (Ab) which binds to the extracellular domain of human Tissue Factor (TF) (SEQ ID NO:810) with a first linker component of a bifunctional linker that comprises two or more linker components followed by sequential addition of the remaining linker component(s) to form an Ab-linker intermediate, and reacting the Ab-linker intermediate with the —NH 2 group of a compound of general Formula I:
wherein:
X is *—C(O)NHCH(CH 2 (R 2 ))— + , wherein * and + represent the respective points of attachment as indicated in Formula I, or X is absent;
R 1 is selected from the group consisting of:
wherein # and % represent the respective points of attachment as indicated in Formula I; and R 2 is phenyl,
to provide the antibody drug conjugate; or
(B) reacting the —NH 2 group on the compound of general Formula I with a first linker component of a bifunctional linker that comprises two or more linker components followed by sequential addition of the remaining linker component(s) to form a linker-toxin intermediate, and reacting the linker-toxin intermediate with a nucleophilic or an electrophilic group on an antigen binding protein (Ab) which binds to the extracellular domain of human Tissue Factor (TF) (SEQ ID NO: 810) to provide the antibody-drug conjugate, wherein, in (A) or (B),
(a) the Ab comprises a VH-CDR1, a VH-CDR2, a VH-CDR3, a VL-CDR1, a VL-CDR2, and a VL-CDR3, wherein
i. the VH-CDR1 comprises SEQ ID NO: 872, the VH-CDR2 comprises SEQ ID NO: 873, the VH-CDR3 comprises SEQ ID NO: 874, the VL-CDR1 comprises SEQ ID NO: 875, the VL-CDR2 comprises SEQ ID NO: 876, and the VL-CDR3 comprises SEQ ID NO: 877,
ii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A3,
iii. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A,
iv. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5,
v. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25A5-T, or
vi. the VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are from the antibody designated 25G1; and
(b) the antibody-drug conjugate comprises one or more moieties represented by Formula IV:
wherein:
X is *—C(O)NHCH(CH 2 (R 2 ))— + , wherein * and + represent the respective points of attachment as indicated in Formula IV, or X is absent;
L is a linker;
! represents the point of attachment of L to the Ab, where L is attached to the Ab through a covalent bond;
R 1 is selected from the group consisting of:
wherein # and % represent the respective points of attachment as indicated in Formula VI; and
R 2 is phenyl.
100 . The process according to claim 98 or claim 99 , wherein the nucleophilic or electrophilic group on the Ab is a thiol or an amine.
101 . The process according to claim 100 , further comprising treating the Ab with a reducing agent to reduce one or more disulfide linkages in the Ab to provide the nucleophilic thiol group.
102 . The process according to any one of claims 98 - 101 , wherein L is represented by:
wherein:
Z represents a functional group that binds to a target group of the Ab;
D represents the point of attachment to the amino group as indicated in Formula I;
Str is a stretcher;
AA 1 and AA 2 are each independently an amino acid, wherein AA 1 -[AA 2 ] m forms a protease cleavage site;
X is a self-immolative group;
s is an integer selected from 0 and 1;
m is an integer selected from the group consisting of 1, 2, 3, and 4; and
o is an integer selected from 0, 1, and 2.
103 . A kit comprising the antibody-drug conjugate of any one of claims 1 - 81 or the pharmaceutical composition of claim 82 , and instructions for use.