Compositions and methods for modification of biomolecules
View Patent ↗The present invention provides modified cycloalkyne compounds; and method of use of such compounds in modifying biomolecules. The present invention features a cycloaddition reaction that can be carried out under physiological conditions. In general, the invention involves reacting a modified cycloalkyne with an azide moiety on a target biomolecule, generating a covalently modified biomolecule. The selectivity of the reaction and its compatibility with aqueous environments provide for its application in vivo (e.g., on the cell surface or intracellularly) and in vitro (e.g., synthesis of peptides and other polymers, production of modified (e.g., labeled) amino acids).
1. A compound of the formula:
wherein:
each of X and X′ is independently:
(a) H;
(b) one or two halogen atoms (e.g., bromo, chloro, fluoro, iodo);
(c) —W—(CH 2 ) n —Z, wherein n is an integer from 1-4; wherein W, if present, is O, N, or S; and wherein Z is nitro, cyano, sulfonic acid, or a halogen;
(d) —(CH 2 ) n —W—(CH 2 ) m —Z, wherein n and m are each independently 1 or 2; wherein W is O, N, S, or sulfonyl; with the proviso that if W is O, N, or S, then Z is nitro, cyano, or halogen; and with the proviso that if W is sulfonyl, then Z is H; or
(e) —(CH 2 ) n —Z, wherein n is an integer from 1-4; and wherein Z is nitro, cyano, sulfonic acid, or a halogen;
Y is H; and
R 1 is selected from a carboxylic acid, an alkyl ester, an aryl ester, a substituted aryl ester, an aldehyde, an amide, an aryl amide, an alkyl halide, a thioester, a sulfonyl ester, an alkyl ketone, an aryl ketone, a substituted aryl ketone, a halosulfonyl, a nitrile, and a nitro.
2. The compound of claim 1 , wherein R 1 is an alkyl ester.
3. The compound of claim 1 , wherein X and X′ are both H, and wherein R 1 is a substituted aryl ester.
4. The compound of claim 3 , wherein R 1 is —O—CH 2 -phenyl-COOH.
5. The compound of claim 1 , wherein X and X′ are both H, and wherein R 1 is —CH 2 -phenyl-COOH.
6. The compound of claim 1 , wherein X is F, wherein X′ is H, and wherein R 1 is —CH 2 -phenyl-COOH.
7. The compound of claim 1 , wherein X is F, wherein X′ is H, and wherein R 1 is —CH 2 -phenyl-CO 2 -methyl.
8. The compound of claim 1 , wherein X′ and X are H, and wherein R 1 is —O—CH 2 —C(═O)—OH.
9. The compound of claim 1 , wherein X and X′ are both fluoro.
10. The compound of claim 1 , wherein X and X′ are both H.
11. The compound of claim 1 , wherein X is two fluoro atoms and X′ is H.