Cross-linkers and their uses
Charged or pro-charged cross-linking moieties and conjugates of cell binding agents and drugs comprising the charged or pro-charged cross-linking moieties and method of making the same.
1. A cross linker represented by formula (I)
wherein:
Y′ represents an amine or thiol reactive functional group that can react with a cell-binding agent;
Q represents a functional group that can react with a cytotoxic drug via a disulfide, thioether, thioester, peptide, hydrazone, ester, ether, carbamate or amide bond;
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , and R 10 are the same or different and are H, linear alkyl having from 1-6 carbon atoms, branched or cyclic alkyl having from 3 to 6 carbon atoms, linear, branched or cyclic alkenyl or alkynyl having from 2 to 6 carbon atoms, a charged substituent selected from anions selected from SO 3 − , X—SO 3 − , OPO 3 2− , X—OPO 3 2− , PO 3 2− , X—PO 3 2− , CO 2 − , and cations selected from a nitrogen containing heterocycle, N + R 11 R 12 R 13 , and X—N + R 11 R 12 R 13 , or a phenyl, wherein:
R 11 , R 12 and R 13 are same or different and are H, linear alkyl having from 1 to 6 carbon atoms, or branched or cyclic alkyl having from 3 to 6 carbon atoms and X represents phenyl or a linear alkyl having from 1 to 6 carbon atoms, or branched or cyclic alkyl having from 3 to 6 carbon atoms;
l, m and n are 0 or an integer from 1 to 4;
A is a phenyl or substituted phenyl, wherein the substituent is a linear alkyl having from 1 to 6 carbon atoms, or a branched or cyclic alkyl having from 3 to 6 carbon atoms, or a charged substituent selected from anions selected from SO 3 − , X—SO 3 − , OPO 3 2− , X—OPO 3 2− , PO 3 2− , X—PO 3 2− , CO 2 —, and cations selected from a nitrogen containing heterocycle, N + R 11 R 12 R 13 , and X—N + R 11 R 12 R 13 , wherein X has the same definition as above, and wherein g is 0 or 1; and
Z is an optional polyethyleneoxy unit of formula (OCH 2 CH 2 ) p , wherein p is 0 or an integer from 2 to about 1000, or F1-E1-P-E2-F2 unit in which E1 and E2 are the same or different and are C═O, O, or NR 14 , wherein R 14 is H, a linear alkyl having from 1-6 carbon atoms, a branched or cyclic alkyl having from 3 to 6 carbon atoms, a linear, branched or cyclic alkenyl or alkynyl having from 2 to 6 carbon atoms; P is a peptide unit between 2 and 20 amino acids in length, wherein E1 or E2 can be linked to the peptide through the terminal nitrogen, terminal carbon or through a side chain of one of the amino acids of the peptide; and F1 and F2 are the same or different and are an optional polyethyleneoxy unit of formula (OCH 2 CH 2 ) p , wherein p is 0 or an integer from 2 to about 1000, provided that when Z is not F1-E1-P-E2-F2, at least one of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , and R 10 is a charged substituent or when g is 1, at least one of A, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , and R 10 is a charged substituent.
2. The cross linker of claim 1 , wherein Y′ is an amine reactive functional group.
3. The cross linker of claim 2 , wherein the amine reactive functional group is selected from the group consisting of an N-hydroxysuccinimide ester, p-nitrophenyl ester, dinitrophenyl ester, and pentafluorophenyl ester.
4. The cross linker of claim 1 , wherein Y′ is a thiol reactive functional group.
5. The cross linker of claim 4 , wherein the thiol reactive functional group is selected from the group consisting of a pyridyldisulfide, nitropyridyldisulfide, maleimide, haloacetate and carboxylic ester.
6. The cross linker of claim 1 , wherein one of R 1 , R 2 , R 3 , R 4 , R 9 and R 10 is a charged substituent selected from SO 3 − , X—SO 3 − , OPO 3 2− , X—OPO 3 2− , N + R 11 R 12 R 13 and X—N + R 11 R 12 R 13 , and the rest are H; l, g and m are each 0; n is 1; and Q and Y′ are each independently, a disulfide, a maleimido, a haloacetyl or a N-hydroxysuccinimide ester.
7. The cross linker of claim 6 , wherein one of R 1 , R 2 , R 3 , R 4 , R 9 and R 10 is SO 3 − or X—SO 3 − , the rest are H; l, g and m are each 0; n is 1; Q is a disulfide, a maleimido or a haloacetyl group; and Y′ is a maleimido group or a N-hydroxysuccinimide ester.
8. The cross linker of claim 7 , wherein Q is a pyridyldithio, nitropyridyldithio, maleimido or haloacetyl group; and Y′ is a N-hydroxysuccinimide ester.
9. The cross linker of claim 1 , wherein Z is an F1-E1-P-E2-F2 unit, wherein E1 and E2 are the same or different and are C═O or NR 14 , wherein R 14 is H, a linear alkyl having 1-6 carbon atoms or a branched or cyclic alkyl having from 3 to 6 carbon atoms; and P is a peptide unit between 2 and 8 amino acids.
10. The cross linker of claim 9 , wherein R 14 is H or a linear alkyl having 1-6 carbon atoms; and P is a peptide unit between 2 and 5 amino acids; and F1 and F2 are the same or different and are optional polyethyleneoxy units of formula (OCH 2 CH 2 ) p , where p is 0 or an integer from 2 to 24.
11. The cross linker of claim 10 , wherein p is 0.
12. The cross linker of claim 11 , wherein P is gly-gly-gly.
13. A cross linker represented by one of the following formulas:
14. The cross linker of claim 6 , wherein Z is absent when Z is the optional polyethylene unit and p=0.
15. The cross linker of claim 13 , wherein the cross linker is represented by the following formula: