Organocatalysts of oxidative protein folding
Catalysts of protein-disulfide isomerization of formula: where R 1 is hydrogen or —COR 4 , where R 4 is an optionally substituted aliphatic group or an optionally substituted aryl group; R 2 is hydrogen or —CO—R 5 , where R 5 is alkyl having 1-8 carbon atoms, an alkenyl having 3-8 carbon atoms or a phenyl, benzyl, phenethyl or naphthyl group; and R 3 is hydrogen or alkyl group having 1-3 carbon atoms. Protein folding buffers comprising one or more of the above compounds. Method of catalyzing, in vivo or in vitro, the isomerization of disulfide linkages in a protein or peptide employing above catalysts. Method of forming, in vivo or in vitro, disulfide linkages in a protein or peptide employing above catalysts.
1. A compound having formula I:
where:
R 1 is hydrogen or a —CO—R 4 group, where R 4 is an optionally substituted aliphatic group or an optionally substituted aryl group;
each R 2 is independently hydrogen or a —CO—R 5 group, where R 5 is an alkyl group having 1-8 carbon atoms, an alkenyl group having 3-8 carbon atoms, a phenyl group, a benzyl group, a phenethyl group or a naphthyl group; and
each R 3 is independently hydrogen or an alkyl group having 1-3 carbon atoms; wherein substituents for R 4 , if present, are selected from halogens, alkyl groups having 1-6 carbon atoms, alkoxy groups having 1-6 carbon atoms and aryl groups having 6-12 carbon atoms.
2. The compound of claim 1 , wherein the calculated log P of the compound (CH 3 ) 2 NR 1 is greater than or equal to 0.
3. The compound of claim 1 , wherein each R 2 and each R 3 are hydrogens.
4. The compound of claim 1 , wherein R 1 is —COR 4 .
5. The compound of claim 4 , wherein R 4 is an alkyl group having 1-20 carbon atoms or an alkenyl group having 3-20 carbon atoms.
6. The compound of claim 4 , wherein R 4 is an alkyl group having 1-6 carbon atoms.
7. The compound of claim 4 , wherein R 4 is an aryl group having 6-12 carbon atoms.
8. The compound of claim 4 , wherein R 4 is a cyclopentyl, cyclohexyl, cyclooctyl, cyclononyl, cyclodecyl, cyclopentenyl, cyclohexenyl, cyclooctenyl, cyclononenyl or cyclodecenyl group.
9. The compound of claim 4 , wherein R 4 is an optionally substituted phenyl, benzyl, biphenyl, naphthyl, indanyl or indenyl group.
10. The compound of claim 4 , wherein R 4 is an alkyl-substituted phenyl group.
11. The compound of claim 1 , wherein R 1 , each R 2 and each of R 3 are hydrogens.
12. The compound of claim 1 , wherein each R 2 and each of R 3 are hydrogens and R 1 is —COR 4 , where R 4 is an unsubstituted straight-chain or branched alkyl group having 3-6 carbon atoms.
13. The compound of claim 1 , wherein each R 2 and each of R 3 are hydrogens and R 1 is —COR 4 , where R 4 is an unsubstituted phenyl group, an unsubstituted benzyl group, an unsubstituted phenethyl group, or an alkyl-substituted phenyl group.
14. The compound of claim 1 , wherein each R 2 and each of R 3 are hydrogens and R 1 is —COR 4 , where R 4 is an unsubstituted cycloalkyl group having 5-8 carbon atoms.
15. The compound of claim 1 , wherein each R 2 is —COR 5 .
16. The compound of claim 1 , wherein each R 2 is —COR 5 , each R 3 is hydrogen and R 1 is —COR 4 .
17. A refolding buffer comprising one or more compounds of claim 1 .
18. A method for forming a disulfide bond in a polypepetide or protein containing at least two cysteines which comprises contacting the polypeptide or protein with one or more catalysts of claim 1 .
19. A method for isomerizing disulfide bonds in a polypeptide or protein containing at least two disulfide bonds which comprises contacting the polypeptide or protein with one or more catalysts of any of claim 1 .
20. The compound of claim 1 , wherein each R 2 and each R 3 are hydrogens and R 1 is a —CO—R 4 group, where R 4 is unsubstituted phenyl or a 4-ethyl phenyl group.
21. The compound of claim 1 , wherein each R 3 is hydrogen, each R 2 is —COR 5 , where R 5 is an alkyl group having 1-3 carbon atoms.
22. The compound of claim 1 , which is
23. The compound of claim 1 , which is
24. A refolding buffer of claim 17 comprising one or both of