Modified antibodies and related compounds, compositions, and methods of use
Provided herein are modified antibodies, compounds used to make them, and intermediates in their synthesis; compositions; formulations and methods, including methods of treating diseases, disorders or conditions, for example, cancer, in humans.
1. A method of modifying an antibody by reacting the antibody with a compound comprising a moiety of the following formula (Ia):
or an enantiomer, diastereomer, or mixtures thereof;
wherein:
each Y and Y′ is independently selected from
wherein:
each R and R′ is independently hydrogen or C 1-6 alkyl, wherein one or more carbons in the C 1-6 alkyl are optionally substituted by an oxo group; or
R and R′, together with the two carbons from the single or double bond to which they are attached, form a saturated or unsaturated carbocyclic ring containing from four to seven ring atoms;
wherein, excluding the two carbons from the single or double bond, one or more of the ring carbon atoms are optionally replaced by N; wherein, excluding the two carbons from the single or double bond, one or more of the ring carbon atoms are optionally substituted by an oxo group; and wherein, if present, the ring nitrogen atom is substituted by a C 1-3 alkyl;
the bond represents a single or a double bond; and
the symbol represents a point of attachment to another group;
wherein R A is selected from the group consisting of hydroxyl, amino, nitro, cyano, chloro, bromo, fluoro, iodo, oxo, carboxyl, C 1-6 alkoxycarbonyl, C 1-6 alkyl, C 2-6 alkenyl, and C 1-6 alkoxy; and
wherein the antibody is modified by reacting the thiols of two cysteine residues from at least one reduced interchain cysteine-cysteine disulfide with the compound.
2. The method of claim 1 , wherein the represents a single bond.
3. A process, which comprises:
(i) reducing one or more interchain cysteine-cysteine disulfide bonds in an antibody with a reducing agent; and
(ii) reacting the free thiol groups from the one or more reduced interchain cysteine-cysteine disulfide bonds with a compound, thus producing a modified antibody;
wherein the compound comprises a moiety of the following formula (la):
or an enantiomer, diastereomer, or mixtures thereof;
wherein:
each Y and Y′ is independently selected from
wherein:
each R and R′ is independently hydrogen or C 1-6 alkyl, wherein one or more carbons in the C 1-6 alkyl are optionally substituted by an oxo group; or
R and R′, together with the two carbons from the single or double bond to which they are attached, form a saturated or unsaturated carbocyclic ring containing from four to seven ring atoms;
wherein, excluding the two carbons from the single or double bond, one or more of the ring carbon atoms are optionally replaced by N; wherein, excluding the two carbons from the single or double bond, one or more of the ring carbon atoms are optionally substituted by an oxo group; and wherein, if present, the ring nitrogen atom is optionally substituted by a C 1-3 alkyl;
the represents a single or a double bond; and
the symbol represents a point of attachment to another group;
wherein R A is selected from the group consisting of hydroxyl, amino, nitro, cyano, chloro, bromo, fluoro, iodo, oxo, carboxyl, C 1-6 alkoxycarbonyl, C 1-6 alkyl, C 2-6 alkenyl, and C 1-6 alkoxy; and
wherein the antibody is modified by reacting the thiols of two cysteine residues from at least one reduced interchain cysteine-cysteine disulfide with the compound.
4. The process of claim 3 , wherein 1 to 4 interchain cysteine-cysteine disulfide bonds are reduced.
5. The process of claim 3 , wherein 4 interchain cysteine-cysteine disulfide bonds are reduced.
6. The process of claim 3 , wherein the reducing agent is dithiothreitol (DTT) or tris(2 carboxyethyl)phosphine (TCEP).
7. The method of claim 1 , wherein the bond represents a double bond.
8. The method of claim 1 , wherein each Y and Y′ is
9. The method of claim 1 , wherein the compound is 1 methyl-3,4-bis(pyridine-2-ylthio)-1H-pyrrole-2,5-dione.
10. The method of claim 1 , wherein the moiety is represented by the following formulas (Ib) or (Ic):
or an enantiomer, diasteriomer, or mixtures thereof;
wherein:
each X and X′ is independently absent or O; W is ═N—; and
each k and k′ is independently an integer of 0, 1, or 2.
11. The method of claim 10 , wherein the moiety is represented by formula (Ib), and wherein:
(a) k and k′ are both 0 and X and X′ are both absent as depicted below:
or
(b) k is 0, X is absent and k′ is 1 as depicted below:
or
(c) k is 1, k′ is 0 and X′ is absent as depicted below:
or
(d) k and k′ are both 1 and X and X′ are both O as depicted below:
or
(e) k and k′ are both 1 and X and X′ are both absent as depicted below:
or
(f) k and k′ are both 1, X is O and X′ is absent as depicted below:
or
(g) k and k′ are both 1, X is absent and X′ is O as depicted below:
12. The method of claim 10 , wherein the moiety is represented by formula (Ic) and wherein:
(a) k is 0, X is absent and k′ is 2 as depicted below:
or
(b) k is 2, k′ is 0 and X′ is absent as depicted below:
or
(c) k and k′ are both 1 and each X and X′ is O as depicted below:
or
(d) k and k′ are both 1, and X and X′ are both absent as depicted below:
or
(e) k is 1, k′ is 2 and X and X′ are both absent as depicted below:
or
(f) k is 2, k′ is 1 and X and X′ are both absent as depicted below:
or
(g) k is 2, k′ is 2 and X and X′ are both absent as depicted below:
or
(h) k and k′ are both 1, X is O and X′ is absent, as depicted below:
or
(i) k and k′ are both 1, X is absent and X′ is O as depicted below:
or
(j) k is 1, k′ is 2, X is O and X′ is absent as depicted below:
or
(k) k is 2, k′ is 1, X is absent and X′ is O as depicted below:
13. The method of claim 12 , wherein in (a), one or both of X′ is also absent as depicted below:
14. The method of claim 12 , wherein in (b), one or both of X is also absent as depicted below:
15. The method of claim 10 , wherein the moiety is of the following formulas (Id) or (le):
wherein:
each R a , R a ′ and R b is C 1-3 alkyl.
16. The method of claim 1 , wherein the moiety is selected from the group consisting of:
and.
17. The method of claim 1 , wherein the moiety is selected from the group consisting of:
and.
18. The process of claim 3 , wherein the bond represents a single bond.
19. The process of claim 3 , wherein the bond represents a double bond.
20. The process of claim 3 , wherein each Y and Y′ is
21. The process of claim 3 , wherein the compound is 1-methyl-3,4-bis(pyridine-2-ylthio)-1H-pyrrole-2,5-dione.
22. The process of claim 3 , wherein the moiety is represented by the following formulas (Ib) or (Ic):
or an enantiomer, diasteriomer, or mixtures thereof;
wherein:
each X and X′ is independently absent or O; W is ═N—; and
each k and k′ is independently an integer of 0, 1, or 2.
23. The process of claim 22 , wherein the moiety is represented by formula (Ib), and wherein:
(a) k and k′ are both 0 and X and X′ are both absent as depicted below:
or
(b) k is 0, X is absent and k′ is 1 as depicted below:
or
(c) k is 1, k′ is 0 and X′ is absent as depicted below:
or
(d) k and k′ are both 1 and X and X′ are both O as depicted below:
or
(e) k and k′ are both 1 and X and X′ are both absent as depicted below:
or
(f) k and k′ are both 1, X is O and X′ is absent as depicted below:
or
(g) k and k′ are both 1, X is absent and X′ is O as depicted below:
24. The process of claim 22 , wherein the moiety is represented by formula (Ic) and wherein:
(a) k is 0, X is absent and k′ is 2 as depicted below:
or
(b) k is 2, k′ is 0 and X′ is absent as depicted below:
or
(c) k and k′ are both 1 and each X and X′ is O as depicted below:
or
(d) k and k′ are both 1, and X and X′ are both absent as depicted below:
or
(e) k is 1, k′ is 2 and X and X′ are both absent as depicted below:
or
(f) k is 2, k′ is 1 and X and X′ are both absent as depicted below:
or
(g) k is 2, k′ is 2 and X and X′ are both absent as depicted below:
or
(h) k and k′ are both 1, X is O and X′ is absent, as depicted below:
or
(i) k and k′ are both 1, X is absent and X′ is O as depicted below:
or
(j) k is 1, k′ is 2, X is O and X′ is absent as depicted below:
or
(k) k is 2, k′ is 1, X is absent and X′ is O as depicted below:
25. The process of claim 24 , wherein in (a), one or both of X′ is also absent as depicted below:
26. The process of claim 24 , wherein in (b), one or both of X is also absent as depicted below:
27. The process of claim 22 , wherein the moiety is represented by the following formulas (Id) or (Ie):
wherein:
each R a , R a ′ and R b is C 1-3 alkyl.
28. The process of claim 3 , wherein the moiety is selected from the group consisting of:
and.
29. The process of claim 3 , wherein the moiety is selected from the group consisting of:
and.