Conjugates of tumor necrosis factor inhibitors to functionalized polymers
This document relates to conjugates of TNF inhibitors or derivatives thereof and functionalized (e.g., mono- or bi-functional) polymers (e.g., polyethylene glycol and related polymers) as well as methods and materials for making and using such conjugates.
1. A method of preparing a compound comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):
or a salt thereof,
wherein:
A is the point of covalent bonding to the terminus of the polymer backbone;
E is O or S;
Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
Z 1 and Z 2 are independently selected from O and NH, wherein only one of Z 1 and Z 2 can be NH;
L is selected from the group consisting of: a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
L 2 is a covalent linking moiety between L on the polymer backbone and B; and
B is a TNF inhibitor or a derivative thereof;
comprising the steps of:
(a) providing a composition comprising a compound of formula (8):
wherein
A is the point of covalent bonding to the terminus of the polymer backbone;
E is O or S;
Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
Z 1 and Z 2 are independently selected from O and NH, wherein only one of Z 1 and Z 2 can be NH;
L is selected from the group consisting of: a divalent radical of a nucleoside, an unsubstituted or substituted alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and an unsubstituted or substituted arylene;
M is a protected group that when deprotected is reactive with a TNF inhibitor or a derivative thereof;
R is a hydrophobic separation handle;
R 1 is absent or a hydrophobic separation handle; and
(b) removing the hydrophobic separation handle(s);
(c) optionally reacting the compound obtained in step (b) with an activating agent; and
(d) reacting the compound obtained in step (b), or, optionally in step (c), with a TNF inhibitor or a derivative thereof.
2. The method of claim 1 , wherein the polymer has from 2 to 100 termini.
3. The method of claim 1 , wherein the polymer backbone has two termini.
4. The method of claim 1 , wherein said polymer backbone is selected from the group consisting of poly(alkylene glycol), poly(oxyethylated polyol), poly(olefinic alcohol), poly(α-hydroxy acid), poly(vinyl alcohol), polyoxazoline, and copolymers.
5. The method of claim 1 , wherein said polymer backbone is poly(ethylene glycol).
6. The method of claim 5 , wherein said poly(ethylene glycol) has an average molecular weight from about 500 Da to about 100,000 Da.
7. The method of claim 1 , wherein said TNF inhibitor is selected from the group consisting of a fusion protein, a monoclonal antibody, and an antibody fragment.
8. The method of claim 1 , wherein said TNF inhibitor is selected from the group consisting of etanercept, infliximab, adalimumab, certolizumab pegol, and golimumab.
9. The method of claim 1 , wherein E is O.
10. The method of claim 1 , wherein E is S.
11. The method of claim 1 , wherein one of Z 1 and Z 2 is NH and the other is O.
12. The method of claim 4 , wherein Z 1 is O and Z 2 is NH.
13. The method of claim 1 , wherein both Z 1 and Z 2 are 0.
14. The method of claim 1 , wherein L is a divalent radical of a nucleoside.
15. The method of claim 1 , wherein L is a substituted or unsubstituted C 1 -C 12 alkylene.
16. The method of claim 1 , wherein R is selected from the group consisting of: trityl, monoalkoxytrityl, dialkoxytrityl, pixyl, alkoxypixyl, fluorenylmethyloxycarbonyl, trifluoroacetyl, acetal, and cyclic acetal.
17. The method of claim 1 , wherein M, when deprotected, is selected from the group consisting of: hydroxyl, amine, thiol, carboxyl, aldehyde, glyoxal, dione, alkenyl, alkynyl, alkedienyl, azide, acrylamide, vinyl sulfone, hydrazide, aminoxy, maleimide, dithiopyridine, and iodoacetamide.
18. The method of claim 1 , wherein the activating agent is selected from pyridinium hydrochloride, pyridinium trifluoroacetate, and buffered carboxylic acids.
19. The method of claim 1 , wherein Y is alkyl.
20. The method of claim 1 , wherein Y is cycloalkyl.