US5169935A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method of making peptides
Assignee: Salk Institute for Biological Studies
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Filing: 1990-06-20
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Granted: 1992-12-08
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Expires: 2010-06-20
Abstract
Abstract Methods for making peptides of a suitable length for solid phase synthesis, which peptides include in their sequence a pair of residues of a different character which have acylated side chains and a residue which has an N-alkylated side chain. A peptide intermediate is constructed on the resin using commercially available starting materials. The N-terminus can be acylated by removing the α-amino protecting group and acylating under standard conditions. First primary amino protecting groups included in those residues to be acylated are removed, and acylation is effected, preferably by using a carboxypyridine or a similar heterocyclic acylating agent. Following such side chain acylation, a second protecting group included in the residue to be N-alkylated is removed, and the N-alkylation reaction is carried out while the peptide remains on the resin using a borohydride and an appropriate aldehyde or ketone. Following cleavage from the resin and removal of any protecting groups still remaining, the peptide is appropriately purified, thus requiring only a single purification to be carried out while forming a synthetic peptide including residues for which the modified amino acids are not readily commercially available.
Inventors
- 👤 Carl A. Hoeger
- 👤 Paula G. Theobald
- 👤 John S. Porter
- 👤 Jean E. F. Rivier
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 23:57
CPC Classifications
C; C07; C07K; C07K1/00; C07K1/06; C07K1/061; C07K1/064; C07K7/00; C07K7/04; C07K7/23; Y; Y02; Y02P; Y02P20/00; Y02P20/50; Y02P20/55
Claims Analysis
Total Claims: 4
Claim 1
Independent Claim
160 words
A method for making a peptide having the formula: Ac-β-D-2NAL-(4C1)D-Phe-D-3PAL-Ser-Lys(nicotinoyl)-D-Lys(nicotinoyl)-Leu-Lys(isopropyl)-Pro-D-Ala-NH 2 ; which method comprises (a) forming a first intermediate peptide by solid phase synthesis having the formula: (N.sup.α Boc)-β-D-2-NAL-(4Cl)D-Phe-D-3PAL-Ser(Bz1)-Lys(Fmoc)-D-Lys(Fmoc)-Leu-Lys(2C1Z)-Pro-D-Ala-NH-[resin support], (b) deblocking the N-terminus by removal of Boc; (c) acetylating said deblocked N-terminus with a molar excess of acetic anhydride in dichloromethane; (d) removing said Fmoc protecting groups on said 5-position Lys and 6-position D-Lys; (e) acylating said deprotected amino groups by reacting with a molar excess of nicotinic acid in DMF, using DIIC as a coupling agent, resulting in a second peptide intermediate having the formula: Ac-β-D-2NAL-(4Cl)D-Phe-D-3PAL-Ser)Bz1)-Lys(nicotinoyl)-D-Lys(Nicotinoyl)-Leu-Lys(2C1Z)-Pro-D-Ala-NH-[resin support]; (f) reacting said second intermediate peptide with cyclohexane in the presence of paladium catalyst on charcoal to accomplish phase transfer hydrogenation at the 2C1Z and Bz1 protecting groups; (g) reacting said deprotected second intermediate peptide with acetone in a solution of sodium borate buffer at a pH of about 9 and (h) cleaving said peptide from the resin with HF.
Claim 2
Independent Claim
160 words
A method for making a peptide having the formula: Ac-β-D-2NAL-(4C1)D-Phe-D-3PAL-Ser-Lys(nicotinoyl)-D-Lys(nicotinoyl)-Leu-Lys(isopropyl)-Pro-D-Ala-NH 2 ; which method comprises (a) forming a first intermediate peptide by solid phase synthesis having the formula: (N.sup.α Boc)-β-D-2-NAL-(4Cl)D-Phe-D-3PAL-Ser(Bz1)-Lys(Fmoc)-D-Lys(Fmoc)-Leu-Lys(2C1Z)-Pro-D-Ala-NH-[resin support], (b) deblocking the N-terminus by removal of Boc; (c) acetylating said deblocked N-terminus with a molar excess of acetic anhydride in dichloromethane; (d) removing said Fmoc protecting groups on said 5-position Lys and 6-position D-Lys; (e) acylating said deprotected amino groups by reacting with a molar excess of nicotinic acid in DMF, using DIIC as a coupling agent, resulting in a second peptide intermediate having the formula: Ac-β-D-2NAL-(4Cl)D-Phe-D-3PAL-Ser)Bz1)-Lys(nicotinoyl)-D-Lys(Nicotinoyl)-Leu-Lys(2C1Z)-Pro-D-Ala-NH-[resin support]; (f) reacting said second intermediate peptide with cyclohexane in the presence of paladium catalyst on charcoal to accomplish phase transfer hydrogenation at the 2C1Z and Bz1 protecting groups; (g) reacting said deprotected second intermediate peptide with acetone in a solution of sodium borate buffer at a pH of about 9 and (h) cleaving said peptide from the resin with HF.
Claim 3
Independent Claim
172 words
A method for making a peptide having the formula: Ac-β-D-2NAL-(4C1)D-Phe-D-3PAL-Ser-Lys(nicotinoyl)-D-Lys(nicotinoyl)-Leu-Lys(isopropyl)-Pro-D-Ala-NH z , which method comprises (a) forming a first intermediate peptide by solid phase synthesis having the formula: (N.sup.α Boc)-β-D-2NAL-(4C1)D-Phe-D-3PAL-Ser(X 3 )-Lys(Fmoc)-D-Lys(Fmoc)-Leu-Lys(2C1Z)-Pro-D-Ala-NH-[resin support], wherein X 3 is a protecting group for a hydroxyl group of Ser; (b) initially deblocking the N-terminus by removal of Boc; (c) acetylating said deblocked N-terminus with a molar excess of acetic anhydride; (d) then removing said Fmoc protecting groups on said 5-position Lys and 6-position D-Lys; (e) acylating said deprotected amino groups by reacting with a molar excess of nicotinic acid plus a coupling agent, resulting in a second peptide intermediate having the formula: Ac-β-D-2NAL-(4C1)D-Phe-D-3PAL-Ser(X 3 )-Lys(nicotinoyl)-D-Lys(Nicotinoyl)-Leu-Lys(2C1Z)-Pro-D-Ala-NH-[resin support]; (f) deprotecting said second intermediate peptide by reacting with cyclohexane in the presence of palladium catalyst on charcoal to accomplish phase transfer hydrogenation at the 2C1Z and X 3 protecting groups; (g) reacting said deprotected second intermediate peptide with acetone in a solution of sodium borate buffer, and (h) then cleaving said peptide from the resin support with HF.
Claim 4
Independent Claim
172 words
A method for making a peptide having the formula: Ac-β-D-2NAL-(4C1)D-Phe-D-3PAL-Ser-Lys(nicotinoyl)-D-Lys(nicotinoyl)-Leu-Lys(isopropyl)-Pro-D-Ala-NH z , which method comprises (a) forming a first intermediate peptide by solid phase synthesis having the formula: (N.sup.α Boc)-β-D-2NAL-(4C1)D-Phe-D-3PAL-Ser(X 3 )-Lys(Fmoc)-D-Lys(Fmoc)-Leu-Lys(2C1Z)-Pro-D-Ala-NH-[resin support], wherein X 3 is a protecting group for a hydroxyl group of Ser; (b) initially deblocking the N-terminus by removal of Boc; (c) acetylating said deblocked N-terminus with a molar excess of acetic anhydride; (d) then removing said Fmoc protecting groups on said 5-position Lys and 6-position D-Lys; (e) acylating said deprotected amino groups by reacting with a molar excess of nicotinic acid plus a coupling agent, resulting in a second peptide intermediate having the formula: Ac-β-D-2NAL-(4C1)D-Phe-D-3PAL-Ser(X 3 )-Lys(nicotinoyl)-D-Lys(Nicotinoyl)-Leu-Lys(2C1Z)-Pro-D-Ala-NH-[resin support]; (f) deprotecting said second intermediate peptide by reacting with cyclohexane in the presence of palladium catalyst on charcoal to accomplish phase transfer hydrogenation at the 2C1Z and X 3 protecting groups; (g) reacting said deprotected second intermediate peptide with acetone in a solution of sodium borate buffer, and (h) then cleaving said peptide from the resin support with HF.
References Cited (Backward Citations)
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