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US 5,516,891 AGranted
Liquid phase synthesis of peptides and peptide derivatives
AssigneeKinerton Limited
InventorsGary A. Siwruk, John S. Eynon
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Abstract
A liquid phase peptide synthetic method which uses (1) Fmoc as the protecting group for the non-side chain amino functionality, (2) ammonia, a primary or secondary amine to remove the Fmoc protecting group, and (3) substituted carbodiimide as the coupling agent for the C to N synthesis of peptides or peptide derivatives in a proper organic solvent.
Claims38
A continuous liquid phase synthetic method for preparing a peptide containing 3-10 amino acid residues with or without modification at the C-terminus or an N-acyl derivative thereof, which method comprises the steps of:
- (a) coupling a first amino acid or a derivative thereof to a second amino acid via an amide bond in an organic solvent containing a substituted carbodiimide, with said first amino acid or derivative thereof having both its non-side chain carboxyl functionality and its side-chain functionalities, if any and necessary, blocked by base-stable groups and said second amino acid with its non-side chain amino functionality blocked by Fmoc and its side chain functionalities, if any and necessary, blocked by base-stable groups;
- (b) detaching said Fmoc with ammonia, a primary amine, or a secondary amine;
- (c) coupling an additional amino acid to the Fmoc-detached coupled product via an amide bond in an organic solvent containing a substituted carbodiimide, with said additional amino acid having its non-side chain amino functionality blocked by Fmoc and its side chain functionalities, if any and necessary, blocked by base-stable groups;
- (d) detaching said Fmoc with ammonia, a primary amine, or a secondary amine;
- (e) repeating steps (c) and (d) until the length of said peptide or said N-acyl derivative is achieved;
- (f) acylating the non-side chain amino group of the Fmoc-detached coupled product from the previous step;
- and
- (g) detaching said base-stable groups;
- wherein said first amino acid or derivative thereof corresponds to a first residue which is the C-terminal residue in said peptide or said N-acyl derivative, said second amino acid corresponds to a second residue adjacent to the first residue in said peptide or said N-acyl derivative, and said additional amino acid corresponds to a third residue adjacent to said second residue in said peptide or said N-acyl derivative;
- providing that when said peptide, rather than said N-acyl derivative, is to be synthesized, step (f) is omitted.
7 more elements
A continuous liquid phase synthetic method for preparing a peptide containing 2-10 amino acid residues with or without modification at the C-terminus or an N-acyl derivative thereof, which method comprises the steps of:
- (a) coupling a first amino acid or a derivative thereof to a second amino acid via an amide bond in an organic solvent containing a substituted carbodiimide, with said first amino acid or derivative thereof having both its non-side chain carboxyl functionality and its side-chain functionalities, if any and necessary, blocked by base-stable groups and said second amino acid with its non-side chain amino functionality blocked by Fmoc and its side chain functionalities, if any and necessary, blocked by base-stable groups;
- (b) detaching said Fmoc with ammonia, a primary amine, or a secondary amine;
- (c) coupling an additional amino acid to the Fmoc-detached coupled product via an amide bond in an organic solvent containing a substituted carbodiimide, with said additional amino acid having its non-side chain amino functionality blocked by Fmoc and its side chain functionalities, if any and necessary, blocked by base-stable groups;
- (d) detaching said Fmoc with ammonia, a primary amine, or a secondary amine;
- (e) repeating steps (c) and (d) until the length of said peptide or said N-acyl derivative is achieved;
- (f) acylating the non-side chain amino group of the Fmoc-detached coupled product from the previous step;
- and
- (g) detaching said base-stable groups;
- wherein said first amino acid or derivative thereof corresponds to a first residue which is the C-terminal residue in said peptide or said N-acyl derivative, said second amino acid corresponds to a second residue adjacent to the first residue in said peptide or said N-acyl derivative, and said additional amino acid corresponds to a third residue adjacent to said second residue in said peptide or said N-acyl derivative;
- providing that when said peptide or said N-acyl derivative contains two amino acid residues, steps (c) through (e) are omitted;
- and further providing that when said peptide, rather than said N-acyl derivative, is to be synthesized, step (f) is omitted;
- and further providing said detached Fmoc is removed by filtration and subsequent washing with an acidic aqueous solution.
9 more elements
Dependencies are inferred from the claim text.