Polypeptide conjugates with extended circulating half-lives
View Patent ↗The present invention relates to compounds and methods for synthesizing compounds wherein the compounds exhibit extended circulating half-life in the blood. The increase in circulating half-life is achieved by conjugating polypeptides to binding groups that exhibit high affinity for human serum albumin.
1. A compound having the formula:
wherein R 1 is (CH 2 ) n , where n is an integer from 0 to 12;
R 2a and R 2b are independently (CH 2 ) m , C(O)NH—(CH 2 ) x , CH 2 —NH—(CH 2 ) y , (CH 2 ) z —C(O), or not present, where m, x, and z are integers from 0 to 12 and y is an integer from 1 to 12;
R 3 is independently C(O)—(CH 2 ) p , where p is an integer of 0 to 12, C(O)CH(CH 3 ), C(O)C(CH 3 ) 2 , or is not present;
R 4 is H, alkyl, or a negative charge and the salts thereof;
X is N, CH, P, or triazine;
W′ is a linear or branched alkyl chain having one or more carbon atoms; and
R 5 is halogen, 1° amine, 2° amine, 3° amine, hydroxyl, thiol, carboxylic acid, sulfonic acid, or activated ester.
2. A compound having the formula:
wherein R 1 is (CH 2 ) n , where n is an integer from 0 to 12;
R 2a and R 2 are independently (CH 2 ) m , C(O)NH—(CH 2 ) x , CH 2 —NH—(CH 2 ) y , (CH 2 ) z —C(O), or not present, where m, x, and z are integers from 0 to 12 and y is an integer from 1 to 12.
R 3 is independently C(O)—(CH 2 ) p , where p is an integer of 0 to 12, C(O)CH(CH 3 ), C(O)C(CH 3 ) 2 , or is not present.
R 4 is H, alkyl, or a negative charge and the salts thereof;
X is N, CH, P, or triazine;
W′ is a linear or branched alkyl chain having one or more carbon atoms; and
R 5 is halogen, 1° amine, 2° amine, 3° amine, hydroxyl, thiol, carboxylic acid, sulfonic acid, or activated ester; and
G is (CH 2 ) q , where q is an integer from 0 to 9, C(O), SO 2 , or S(O).
3. The compound of claims 1 or 2 , wherein one or more of said carbon atoms of W′ are substituted independently with cycloalkane, a heteroatom, a substituted heteroatom, CO—O, CO—NH, CO—S, CO, SO 2 , SO 2 —NH, or SO 2 —O.
4. The compound of claim 3 , wherein the substituted heteroatom is selected from the group consisting of N-(alkyl), N-(aryl), N-(alkyl-aryl), phosphine, phosphate, thiophosphate, phosphodiester, and aminophosphate.
5. The compound according to claim 1 or 2 , wherein the aryl moieties are independently
wherein R is selected from the group consisting of H, OCH 3 , halogen, CN, NO 2 , CF 3 , OCF3, OH, O(C 1 –C 4 )Alkyl, SH, S(C 1 –C 4 )Alkyl, SO(C 1 –C 4 )Alkyl, SO 2 (C 1 –C 4 )Alkyl, SO3H, SO 2 NH 2 , SO 2 NH(C 1 –C 4 )Alkyl, NH 2 , NH(C 1 –C 4 )Alkyl, N((C 1 –C 4 )Alkyl) 2 , CO 2 H, (C 1 –C 6 )Alkyl, and (C 3 –C 8 )Cycloalkyl.
6. The compound according to claim 5 , wherein the aryl moiety is selected from the group consisting of:
7. A conjugated polypeptide having the general Formula I,
wherein R 2a and R 2b are independently (CH 2 ) m , C(O)NH—(CH 2 ) x , CH 2 —NH—(CH 2 ) y , (CH 2 ) z —C(O), or not present, where m, x, and z are integers from 0 to 12 and y is an integer from 1 to 12;
R 3 is independently C(O)—(CH 2 ) p , where p is an integer of 0 to 12, C(O)CH(CH 3 ), C(O)C(CH 3 ) 2 , or is not present;
X is N, CH, P, or triazine;
W is a linear or branched alkyl chain having one or more carbon atoms;
Z is independently NH, O, S, S(O), CH 2 , NH—CO, SO 2 , CH—R, C(R) 2 , CH═CH, CH═N, wherein R is alkyl or aryl; and
wherein the conjugated polypeptide specifically binds to albumin.
8. A conjugated polypeptide having the general Formula II:
wherein R 2a , R 2b , and R 3 independently comprise a linear chain of 0 to 14 atoms;
X is one atom or 5 or more atoms, wherein the 5 or more atoms form a cyclic or a heterocyclic ring structure;
W is a linker; and
G is a chain of 0 to 9 carbon atoms,
wherein the conjugated polypeptide specifically binds to albumin.
9. The conjugated polypeptide of claim 8 , wherein R 2a and R 2b are independently (CH 2 ) m , C(O)NH—(CH 2 ) x , CH 2 —NH—(CH 2 ) y , (CH 2 ) z —C(O), or not present, where m, x, and z are integers from 0 to 12 and y is an integer from 1 to 12.
10. The conjugated polypeptide of claim 8 or 9 , wherein R 3 is independently C(O)—(CH 2 ) p , where p is an integer of 0 to 12, C(O)CH(CH 3 ), C(O)C(CH 3 ) 2 , or is not present.
11. The conjugated polypeptide of any one of claims 8 or 9 , wherein X is N, CH, P, or triazine.
12. The conjugated polypeptide of any one of claims 8 or 9 , wherein G is (CH 2 ) q , where q is an integer from 0 to 9, or C(O).
13. The conjugated polypeptide of claims 7 or 8 , wherein the conjugate comprises a therapeutic polypeptide selected from the group consisting of erythropoietin, insulin, interferon α-2b, interferon β, interferon γ, granulocyte colony stimulating factor, human growth hormone, granulocyte macrophage colony stimulating factor, relaxin, urokinase, streptokinase, tissue plasminogen activator, and tumor necrosis factor.
14. The conjugated polypeptide according to claims 7 or 8 , wherein the conjugate specifically binds to site II of albumin.
15. The conjugated polypeptide according to claims 7 or 8 , wherein the conjugate exhibits a serum half-life that is greater than that of the corresponding unconjugated polypeptide.
16. The conjugated polypeptide of claim 8 , wherein W comprises a structural element having the formula,
wherein R 1 is (CH 2 ) n , where n is an integer from 0 to 12; and
R 4 is H, alkyl, or a negative charge and the salts thereof.
17. The conjugated polypeptide of claim 8 , wherein W comprises a structural element having the formula,
wherein R 1 is (CH 2 ) n , and where n is an integer from 0 to 12.
18. The conjugated polypeptide of claim 8 , wherein W comprises a linear or branched alkyl chain having one or more carbon atoms.
19. The conjugated polypeptide of claims 7 or 18 , wherein one or more of said carbon atoms of W are substituted independently with cycloalkane, a heteroatom, a substituted heteroatom, CO—O, CO—NH, CO—S, CO, SO 2 , SO 2 —NH, or SO 2 —O.
20. The conjugated polypeptide of claim 19 , wherein the substituted heteroatom is selected from the group consisting of N(alkyl), N(aryl), and N(alkyl-aryl), phosphine, phosphate, thiophosphate, phosphodiester, and aminophosphate.
21. The conjugated polypeptide of claims 7 or 8 , wherein the aryl moieties are independently
wherein R is selected from the group consisting of H, OCH 3 , halogen, CN, NO 2 , CF 3 , OCF 3 , OH, O(C 1 –C 4 )alkyl, SH, S(C 1 –C 4 )alkyl, SO(C 1 –C 4 )alkyl, SO 2 (C 1 –C 4 )alkyl, SO 3 H, SO 2 NH 2 , SO 2 NH(C 1 –C 4 )alkyl, NH 2 , NH(C 1 –C 4 )alkyl, N((C 1 –C 4 )alkyl) 2 , CO 2 H, (C 1 –C 6 )alkyl, and (C 3 –C 8 )cycloalkyl.
22. The conjugated polypeptide according to claim 21 , wherein the aryl moiety is selected from the group consisting of:
23. The compound according to claim 1 , wherein the activated ester is selected from the group consisting of acid chloride, acid anhydride, maleimide, aldehyde, and N-hydroxysuccinimide (NHS) ester.
24. A conjugated polypeptide having the general Formula III:
wherein R 4 is H, alkyl, or a negative charge and the salts thereof;
R 6 comprises a binding moiety containing one or more carbon atoms;
W is a linear or branched alkyl chain having one or more carbon atoms;
wherein the conjugated polypeptide binds albumin, and comprises a therapeutic polypeptide selected from the group consisting of erythropoietin, insulin, interferon α-2b, interferon β, interferon γ, granulocyte colony stimulating factor, human growth hormone, granulocyte macrophage colony stimulating factor, relaxin, urokinase, streptokinase, tissue plasminogen activator, and tumor necrosis factor.
25. The conjugated polypeptide of claim 24 , wherein one or more of said carbon atoms of W are substituted independently with cycloalkane, a heteroatom, a substituted heteroatom, CO—O, CO—NH, CO—S, CO, SO 2 , SO 2 —NH, or SO 2 —O.
26. The conjugated polypeptide of claim 25 , wherein the substituted heteroatom is selected from the group consisting of N-(Alkyl), N-(Aryl), and N-(Alkyl-Aryl), phosphine, phosphate, thiophosphate, phosphodiester, and aminophosphate.
27. The conjugated polypeptide of claim 26 , wherein R 6 further comprises at least one reactive moiety selected from the group consisting of halogen, 1° amine, 2° amine, 3° amine, hydroxyl, thiol, carboxylic acid, sulfonic acid, and activated ester.
28. The conjugated polypeptide of claim 27 , wherein the activated ester is selected from the group consisting of acid chloride, acid anhydride, maleimide, aldehyde, and N-hydroxysuccinimide (NHS) ester.
29. The compound according to claim 1 , wherein n is 1, R 2a is CH 2 —NH—(CH 2 ) 2 , R 2b is not present, R 4 is H; and X is CH.
30. The compound according to claim 29 , wherein the aryl moiety is
31. The compound according to claim 1 , wherein n is 1, R 2a is C(O)—NH—(CH 2 ) 2 , R 2b is not present, R 4 is H; and X is CH.
32. The compound according to claim 1 , wherein the aryl moiety is selected from the group consisting of
33. The compound according to claim 1 , wherein n is 1, R 2a is CH 2 , R 2b is not present, R 4 is H, and X is CH.
34. The compound according to claim 1 , wherein the aryl moiety is selected from the group consisting of:
35. A method for increasing the half life of a polypeptide, the method comprising conjugating a therapeutic polypeptide to the compound of claim 1 or claim 2 .
36. The compound of claim 2 , wherein if said X is triazine, said triazine forms a bicyclic ring structure together with the ring defined by R 2a , R 2b , the two Ns, and the G.
37. The conjugated polypeptide of claim 8 , wherein said cyclic or heterocyclic ring structure of X forms a bicyclic ring structure together with the ring defined by R 2a , R 2b , the two Ns, and the G.
38. A compound having the formula:
wherein R 1 is (CH 2 ) n , where n is an integer from 0 to 12;
R 2a and R 2b are independently (CH 2 ) m , C(O)NH—(CH 2 ) x , CH 2 —NH—(CH 2 ) y , (CH 2 ) z —C(O), or not present, where m, x, and z are integers from 0 to 12 and y is an integer from to 12;
R 3 is independently C(O)—(CH 2 ) p , where p is an integer of 0 to 12, C(O)CH(CH 3 ), C(O)C(CH 3 ) 2 , or is not present;
R 4 is H, alkyl, or a negative charge and the salts thereof;
X is N, CH, P, or triazine;
W′ is a linear or branched alkyl chain having one or more carbon atoms;
R 5 is halogen, 1° amine, 2° amine, 3° amine, hydroxyl, thiol, carboxylic acid, sulfonic acid, or activated ester; and
C 1-10 comprises a non-aromatic moiety having 1–10 carbon atoms.
39. A compound having the formula:
wherein R 1 is (CH 2 ) n , where n is an integer from 0 to 12;
R 2a and R 2b are independently (CH 2 ) m , C(O)NH—(CH 2 ) x , CH 2 —NH—(CH 2 ) y , (CH 2 ) z —C(O), or not present, where m, x, and z are integers from 0 to 12 and y is an integer from 1 to 12;
R 3 is independently C(O)—(CH 2 ) p , where p is an integer of 0 to 12, C(O)CH(CH 3 ), C(O)C(CH 3 ) 2 , or is not present;
R 4 is H, alkyl, or a negative charge and the salts thereof;
X is N, CH, P, or triazine;
W′ is a linear or branched alkyl chain having one or more carbon atoms;
R 5 is halogen, 1° amine, 2° amine, 3° amine, hydroxyl, thiol, carboxylic acid, sulfonic acid, or activated ester; and
C 1-10 comprises a non-aromatic moiety having 1 10 carbon atoms.
40. A compound having the formula:
wherein R 1 is (CH 2 ) n , where n is an integer from 0 to 12;
R 2a and R 2b are independently (CH 2 ) m , C(O)NH—(CH 2 ) x , CH 2 —NH—CH 2 ) y , (CH 2 ) z —C(O), or not present, where m, x, and z are integers from 0 to 12 and y is an integer from 1 to 12.
R 3 is independently C(O)—(CH 2 ) p , where p is an integer of 0 to 12, C(O)CH(CH 3 ), C(O)C(CH 3 ) 2 , or is not present.
R 4 is H, alkyl, or a negative charge and the salts thereof;
X is N, CH, P, or triazine;
W′ is a linear or branched alkyl chain having one or more carbon atoms;
R 5 is halogen, 1° amine, 2° amine, 3° amine, hydroxyl, thiol, carboxylic acid, sulfonic acid, or activated ester;
G is (CH 2 ) q , where q is an integer from 0 to 9, C(O), SO 2 , or S(O); and
C 1-10 comprises a non-aromatic moiety having 1–10 carbon atoms.
41. A compound having the formula:
wherein R 1 is (CH 2 ) n , where n is an integer from 0 to 12;
R 2a and R 2b are independently (CH 2 ) m , C(O)NH—(CH 2 ) x , CH 2 —NH—(CH 2 ) y , (CH 2 ) z —C(O), or not present, where m, x, and z are integers from 0 to 12 and y is an integer from 1 to 12.
R 3 is independently C(O)—(CH 2 ) p , where p is an integer of 0 to 12, C(O)CH(CH 3 ), C(O)C(CH 3 ) 2 , or is not present.
R 4 is H, alkyl, or a negative charge and the salts thereof;
X is N, CH, P, or triazine;
W′ is a linear or branched alkyl chain having one or more carbon atoms;
R 5 is halogen, 1° amine, 2° amine, 3° amine, hydroxyl, thiol, carboxylic acid, sulfonic acid, or activated ester;
G is (CH 2 ) q , where q is an integer from 0 to 9, C(O), SO 2 , or S(O); and
C 1-10 comprises a non-aromatic moiety having 1–10 carbon atoms.
42. The compound of any one of claims 38 , 39 , 40 , or 41 , wherein the non-aromatic moiety comprises a linear, branched, or cyclic alkyl, alkenyl, or alkynyl moiety.
43. The compound of claim 42 , wherein the cyclic moiety comprises one or more heteroatoms.
44. The compound of claim 43 , wherein the heteroatom is selected from the group consisting of N, O, and S.