Epimerization-free N to C solid-phase peptide synthesis

Patent metadata

Field	Value
Publication number	US-11427612-B2
Application number	US-201916546549-A
Country	US
Kind code	B2
Filing date	Aug 21, 2019
Priority date	Aug 21, 2018
Publication date	Aug 30, 2022
Grant date	Aug 30, 2022

Abstract

The present disclosure provides a method of solid-phase peptide synthesis from the N terminus to C terminus without detectable epimerization of the C-terminal amino acid. The method includes using derivatized amino acids comprising a diamino-aryl group.

First claim

The invention claimed is:

1. A method of synthesizing a peptide, wherein the method comprises preparing a first amino acid for synthesis, coupling an amino group of a second amino acid to the carboxyl group of the first amino acid to obtain a peptide, and optionally elongating the peptide by coupling one or more amino acids sequentially to a C-terminus of the peptide, wherein the amino acids used in the method are derivatized amino acids comprising a diamino-aryl group, and wherein the method proceeds without detectable epimerization of the second or sequentially coupled amino acid at the carboxyl terminus of the peptide.

2. The method of claim 1 , wherein the method comprises solid-phase peptide synthesis, solution phase peptide synthesis, or fluorous phase peptide synthesis.

3. The method of claim 1 , wherein the method comprises derivatizing amino acids by synthesizing amino acid diaminobenzoyl derivatives.

4. The method of claim 3 , wherein synthesis of amino acid diaminobenzoyl derivatives comprises obtaining Fmoc protected amino acid (Fmoc-AA-OH), reacting diaminobenzoylOMe (DbzOMe) with Fmoc-AA-OH to obtain Fmoc-AA-DbzOMe, and reacting with piperidine to remove Fmoc to yield H-AA-DbzOMe derivative.

5. The method of claim 1 , wherein synthesis of amino acid diamino-aryl derivatives comprises obtaining Fmoc protected amino acid (Fmoc-AA-OH), reacting diamino-aryl molecule with Fmoc-AA-OH to obtain Fmoc-AA-diamino-aryl molecule, and reacting with piperidine to remove Fmoc to yield H-AA-diamino aryl derivative.

6. The method of claim 1 , wherein preparing the first amino acid comprises attaching the first amino acid to a resin prior to coupling the amino group of the second amino acid to the carboxyl group of the first amino acid.

7. The method of claim 6 , wherein attaching the first amino acid to the resin comprises anchoring the α-amino group or a side-chain of the first amino acid to the resin.

8. The method of claim 7 , wherein the method further comprises activating the first amino acid to form a first amino acid comprising an N-acyl urea group.

9. The method of claim 8 , wherein activating the first amino acid comprises treating with 4-nitrophenyl chloroformate or other phosgene equivalent and followed by treating with Hünig's base.

10. The method of claim 9 , wherein coupling comprises adding a derivatized second amino acid to displace the N-acyl urea group on the first amino acid at its C terminus to yield a peptide.

11. The method of claim 10 , wherein the method further comprises elongation of the peptide comprising repetition of activation of the peptide and coupling of the peptide with another derivatized amino acid to obtain an N-acyl urea group terminated peptide.

12. The method of claim 11 , wherein the method further comprises after elongating the peptide to a desired length, cleaving the N-acyl urea group from the C-terminus of the peptide to obtain a C-terminally functionalized unprotected peptide or a C-terminally functionalized protected peptide.

13. The method of claim 12 , wherein cleaving the peptide comprises acidic resin cleavage to obtain a C-terminally functionalized unprotected peptide.

14. The method of claim 13 , wherein the acidic resin cleavage comprises treatment with trifluoroacetic acid (TFA) to cleave the peptide from the resin.

15. The method of claim 12 , wherein cleaving the peptide comprises nucleophilic resin cleavage to obtain a C-terminally functionalized protected peptide.

16. The method of claim 15 , wherein nucleophilic resin cleavage comprises treatment with one of the following nucleophiles: NH 3 , BuNH 2 , H 2 N(CH 2 ) 3 N 3 , propargylamine, aniline, H 2 NNH 2 , MeHNOMe, MeOH, EtOH, i-PrOH, BnOH, PhOH, H 2 O, or NaBH 4 .

17. The method of claim 11 , wherein unreacted derivatized amino acids are recovered and reused.

18. The method of claim 10 , wherein the method further comprises elongation of the peptide in the C to N direction.

19. The method of claim 18 , wherein elongation to the desired length is completed in the N to C direction.

20. The method of claim 1 , wherein the method proceeds without detectable epimerization of the second and sequentially coupled amino acid at the carboxyl terminus of the peptide.

Assignees

• Univ Wayne State

Inventors

• Stockdill Jennifer L
• Arbour Christine A

Classifications

• C07K1/084

  containing nitrogen · CPC title

• C07K1/023

  using racemisation inhibiting agents · CPC title

• C07K1/10

  using coupling agents {(C07K1/006 takes precedence)} · CPC title

• C07K1/042 Primary

  characterised by the nature of the carrier · CPC title

• C07K1/062 Primary

  for alpha- or omega-carboxy functions · CPC title