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US4086136A
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Process for producing a peptide using a serine or thiol proteinase
Assignee: Sagami Chemical Research Institute
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Filing: 1976-10-14
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Granted: 1978-04-25
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Expires: 1996-10-14
Abstract
Abstract A peptide having the formula X--A--B--Y-- wherein A and B are the same or different and each represents an amino acid residue or a peptide residue, X represents an amino protective group, Y represents a carboxyl protective group selected from the group consisting of tertiary alkoxy, and benzyloxy, benzylamino and benzhydrylamino which can be substituted with an inert substituent, is prepared by a process which comprises reacting an amino acid or peptide having an N-terminal protective group, or a salt thereof of the formula: X--A--OH with an amino acid or peptide having a C-terminal protective group or a salt thereof of the formula: H--B--Y in the presence of a thiol proteinase or serine proteinase enzyme in an aqueous solution having a pH sufficient to maintain the enzyme activity of said thiol proteinase or serine proteinase.
Inventors
- 👤 Yoshikazu Isowa
- 👤 Muneki Ohmori
- 👤 Hideaki Kurita
- 👤 Tetsuya Ichikawa
- 👤 Masanari Sato
- 👤 Kaoru Mori
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 15:53
CPC Classifications
C; C07; C07K; C07K5/00; C07K5/04; C07K5/06; C07K5/06086; C07K5/06095; C07K1/00; C07K1/02; C07K5/08; C07K5/0819; C12; C12P; C12P21/00; C12P21/02; Y; Y02; Y02P; Y02P20/00; Y02P20/50; Y02P20/55
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
141 words
A process for producing a peptide having the formula X--A--B--Y wherein A and B are the same or different and represent an amino acid residue or a peptide residue; X represents an amino protective group; and Y represents a carboxyl protective group selected from the group consisting of tertiary alkoxy, and benzyloxy, benzylamino and benzhydrylamino which can be substituted with an inert substituent, which comprises: reacting an acid component of an amino acid or peptide having N-terminal protective group or a salt thereof of the formula X-A-OH with an amino component of an amino acid or peptide having a C-terminal protective group or a salt thereof of the formula H--B--Y in the presence of a thiol proteinase or serine proteinase enzyme in an aqueous solution having a pH sufficient to maintain the enzyme activity of said thiol proteinase or serine proteinase.
Claim 2
Dependent Claim
141 words
A process for producing a peptide having the formula X--A--B--Y wherein A and B are the same or different and represent an amino acid residue or a peptide residue; X represents an amino protective group; and Y represents a carboxyl protective group selected from the group consisting of tertiary alkoxy, and benzyloxy, benzylamino and benzhydrylamino which can be substituted with an inert substituent, which comprises: reacting an acid component of an amino acid or peptide having N-terminal protective group or a salt thereof of the formula X-A-OH with an amino component of an amino acid or peptide having a C-terminal protective group or a salt thereof of the formula H--B--Y in the presence of a thiol proteinase or serine proteinase enzyme in an aqueous solution having a pH sufficient to maintain the enzyme activity of said thiol proteinase or serine proteinase.
Claim 3
Dependent Claim
33 words
The process of claim 1, wherein said thiol proteinase is papain, Stembromelein, Ficin, Cathepsin B, Chymopapain, or Streptococcal proteinase and said serine proteinase is subtilisin, Aspergillus alkaline proteinase, Elastase, α-Lytic proteinase or Chymotryspin.
Claim 4
Dependent Claim
45 words
The process of claim 1, wherein the pH is maintained at a sufficient level by reacting the amino acid or peptide reactants in a buffer solution at a pH of 4 to 7.5 for the thiol proteinase and 6 to 9 for the serine proteinase.
Claim 5
Dependent Claim
42 words
The process of claim 1, wherein the pH is maintained at the desired level by detecting the pH of the reaction mixture and adding an acid or a base to said aqueous solution depending upon the measured pH of the reaction mixture.
Claim 6
Dependent Claim
29 words
The process of claim 1, wherein the reaction is conducted with a reactant ratio of 0.8 to 2 moles of the acid component per mole of the amine component.
Claim 7
Dependent Claim
32 words
The process of claim 1, wherein the reaction is conducted by adding 10 to 500 mg of said thiol proteinase or serine proteinase to said solution per mmole of said amine component.
Claim 8
Dependent Claim
69 words
The process of claim 1, wherein the N-terminal protective group of the acid component is a tertiary alkoxycarbonyl group or a benzyloxycarbonyl group which can be substituted with an inert substituent, p-toluenesulfonyl or o-nitrophenyl sulfenyl, and the C-terminal protective group of the amine component is tertiary alkoxy, benzyloxy (--OBzl), p-nitrobenzyloxy [--OBzl (p-NO 2 )], benzhydryloxy, (--OBzh), benzylamino (--NHBzl), 2,4-dimethoxybenzylamino (--NHDMB), benzhydrylamino(--NHBzh) which can be substituted with an inert substituent.
Claim 9
Dependent Claim
40 words
The process of claim 7, wherein said tert-alkoxycarbonyl group is t-butyloxycarbonyl, or t-amyloxycarbonyl; said substituted benzyloxycarbonyl is p-methoxybenzyloxycarbonyl, 3-5-dimethoxybenzyloxycarbonyl, p-phenylazobenzyloxycarbonyl, or 2,4,6-trimethylbenzyloxycarbonyl, said tertiary alkoxy group is t-butoxy, and said inert substituent of said benzyhydrylamino group is 2,4-dimethoxyoxybenzylamino or benzyhydrylamino.
Claim 10
Dependent Claim
56 words
The process of claim 1, wherein A and B are the same or different and represent an amino acid residue or peptide residue wherein the amino acid is an aliphatic amino acid, an oxyamino acid, a sulfur-containing amino acid, a monoamino dicarboxylic acid, a diamino monocarboxylic acid, an aromatic amino acid or a heterocyclic amino acid.
Claim 11
Dependent Claim
80 words
The process of claim 9, wherein said aliphatic amino acid is a monoaminomonocarboxylic selected from the group consisting of glycine, alanine, valine, norvaline, leucine, isoleucine, and norleucine, said oxyamino acid is serine, threonine or homoserine; said sulfur-containing amino acid is methionine, cystine or cysteine; said monoamino dicarboxylic acid is aspartic acid or glutamic acid; said diamino monocarboxylic acid is ornithine, lysine, or arginine; said aromatic amino acid is phenylalanine or tyrosine, and said heterocyclic amino acid is histidine or tryptophan.
References Cited (Backward Citations)
US3972773A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1981-04-24 | 1981-10-24 | 1982-04-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-04-24 | 1985-10-24 | 1986-04-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-04-24 | 1989-10-24 | 1990-04-25 | $7700.00 | Window Closed / Lapsed |