Ligation method and reagents to form an amide bond
AssigneeWisconsin Alumni Research Foundation
InventorsRonald T. Raines, Laura L. Kiessling, Bradley L. Nilsson
Life of this patent
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Abstract
Methods and reagents for the formation of amide bonds between an activated carboxylic acid derivative and an azide useful in the synthesis of peptides, proteins and derivatized or labeled amino acids, peptide or proteins. The method involves the formation of a phosphinothioester which reacts with an azide resulting in amide formation. The invention provides phosphinothiol reagents which convert activated carboxylic acid derivatives to phosphinothioesters which then react with azides to form an amide bond. The methods and reagents of the invention can be used for stepwise synthesis of peptides on solid supports or for the ligation to two or more amino acids, two or more peptide or two or more protein fragments.
Claims58
A method for forming an amide bond which comprises the step of reacting a phosphinothioester with an azide followed by hydrolysis of combined reactants to form an amide bond.
A phosphinothiol reagent having the formula:
- where:
- n and m are 0 or integers equal to 1-3, inclusive, and n+m=0-4, inclusive;
- the dashed line indicates that a double bond may be present or that the bond may be part of an aromatic group, and R4, R6 and R8 are not present if there is a double bond between the carbons or the bond is a part of an aromatic ring;
- R1 and R2 are groups independently selected from aliphatic, alicyclic, heteroalicyclic, aromatic, or heteroaromatic groups which are optionally substituted, wherein substituents are selected from the group consisting of halides, and OH, OR, COH, COR, COOH, COOR, and N(R′)2 groups, where R, independent of other R, is an aliphatic, alicyclic, heteroalicyclic, aromatic or heteroaromatic group and each R′, independent of other R′ is a hydrogen, or an aliphatic, alicyclic, heteroalicyclic, aromatic or heteroaromatic group, R and R′ are optionally substituted as listed above for R1 and R2;
- and R1 and R2, one or more non-neighboring CH2 groups can be replaced with O, S, GO, COO, or CONR′;
- and
- R3-R8, independently, are selected from hydrogens, or aliphatic, alicyclic, heteroalicyclic, aromatic or heteroaromatic groups which are optionally substituted as listed for R1 and R2, in R3-R8 one or more non-neighboring CH2 groups can be replaced with O, S, CO, COO, or CONR′ groups where R′ is defined as for R1 and R2, R1 and R2 are optionally covalently linked to form a cyclic group, including a bicyclic group, and two or more of R3-R8 are optionally covalently linked to form a cyclic group, including a bicyclic group wherein the reagent is not o-(diphenylphosphino)benzenethiol.
Dependencies are inferred from the claim text.