US4569967A
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Synthesis of N-substituted peptide amides
Assignee: Salk Institute for Biological Studies
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Filing: 1983-10-24
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Granted: 1986-02-11
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Expires: 2003-10-24
Abstract
Abstract Peptide N-alkylamides, and other C-terminal N-substituted amides, can be synthesized using solid-phase synthesis on a benzene-containing resin which is suitably methylated. Reactive amino groups are attached directly or indirectly to the methyl groups, for example, such as by reacting commercially available chloromethylated polystyrene resins with an alkylamine to create a resin-amine. The C-terminal amino acid of the desired peptide is linked to the resin-amine via an amide linkage, and the peptide is thereafter built in normal fashion. Treatment of the completed peptide intermediate with HF is effective to both effect deprotection and cleave the peptide from the resin in the form of the N-substituted amide. Examples include peptide N-ethylamides, N-fluoroethylamide, N-anilides and other substituted N-benzylamides.
Inventors
- 👤 Wayne D. Kornreich
- 👤 Harry A. Anderson
- 👤 John S. Porter
- 👤 Jean E. F. Rivier
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-28 12:20
CPC Classifications
C; C07; C07K; C07K5/00; C07K5/04; C07K5/08; C07K5/0821; C07K5/0823; C07K1/00; C07K1/04
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
108 words
A method for synthesizing a peptide having a substitute amide at its C-terminus using solid phase synthesis, which method comprises providing a synthetic resin having a plurality of p-benzyl moieties, the methyl groups of which are substituted only with -NHQ, wherein Q is lower alkyl, lower fluoroalkyl, phenyl or substituted phenyl, reacting said synthetic resin with an amino acid having its alpha-amino group blocked to cause coupling of the carboxyl group thereof to said substituent by a substituted amide linkage, deblocking said alpha-amino group, repeating said reacting step to create a peptide and cleaving said peptide from said resin so that the C-terminus of said peptide is --CONHQ.
Claim 2
Independent Claim
108 words
A method for synthesizing a peptide having a substitute amide at its C-terminus using solid phase synthesis, which method comprises providing a synthetic resin having a plurality of p-benzyl moieties, the methyl groups of which are substituted only with -NHQ, wherein Q is lower alkyl, lower fluoroalkyl, phenyl or substituted phenyl, reacting said synthetic resin with an amino acid having its alpha-amino group blocked to cause coupling of the carboxyl group thereof to said substituent by a substituted amide linkage, deblocking said alpha-amino group, repeating said reacting step to create a peptide and cleaving said peptide from said resin so that the C-terminus of said peptide is --CONHQ.
Claim 3
Dependent Claim
15 words
A method in accordance with claim 1 wherein said resin is a cross-linked polystyrene resin.
Claim 4
Dependent Claim
15 words
A method in accordance with claim 1 wherein said cleaving is carried out with HF.
Claim 5
Dependent Claim
17 words
A method in accordance with claim 1 wherein Q is --C 6 H 4 NH 2 .
Claim 6
Dependent Claim
22 words
A method in accordance with claim 4 wherein said peptide is cleaved as a N-aminoanilide which is subsequently oxidized to a N-nitroanilide.
Claim 7
Dependent Claim
22 words
A method in accordance with claim 5 wherein said oxidation is effected by treating with H 2 O 2 under acidic conditions.
Claim 8
Dependent Claim
22 words
A method in accordance with claim 5 wherein the C-terminus of said peptide is ##STR1## which is oxidized to create a para-nitroanilide.
Claim 9
Dependent Claim
21 words
A method in accordance with claim 6 wherein HCl is used to provide said acidic conditions and said peptide is ##STR2##
Claim 10
Dependent Claim
15 words
A method in accordance with claim 1 wherein Q is --CH 2 CH 3 .
Claim 11
Dependent Claim
15 words
A method in accordance with claim 1 wherein Q is --CH 2 CF 3 .
Claim 12
Dependent Claim
15 words
A method in accordance with claim 1 wherein Q is --CH 2 CHF 2 .
Claim 13
Dependent Claim
17 words
A method in accordance with claim 1 wherein Q is --CH 2 CH 2 CH 3 .
Claim 14
Dependent Claim
13 words
A method in accordance with claim 1 wherein Q is --CH 3 .
Claim 15
Dependent Claim
15 words
A method in accordance with claim 1 wherein Q is CHCH 3 CH 3 .
Claim 16
Dependent Claim
33 words
A method in accordance with claim 4 wherein a chloromethylated polystyrene resin is reacted with p-aminoaniline in DMF at about 60°-100° C. for at least about 16 hours to form said substituted resin.
Claim 17
Dependent Claim
28 words
A method in accordance with claim 9 wherein a chloromethylated polystyrene resin is reacted with ethylamine at about 0° to about 5° C. to form said substituted resin.
Claim 18
Independent Claim
163 words
A method for synthesizing a peptide containing at least one Asp or Glu residue within its chain and having a substituted amide at its C-terminus using solid phase synthesis, which method comprises providing a synthetic resin having a plurality of substituted p-benzl moieties of the formula --C 6 H 4 CH 2 NHQ wherein Q is lower alkyl, lower fluoroalkyl, phenyl or substituted phenyl, reacting said synthetic resin with an amino acid having its alpha-amino group blocked to cause coupling of the carboxyl group thereof to the nitrogen of said moiety by a substituted amide linkage, deblocking said alpha-amino group, repeating said reacting and deblocking steps with such additional amino acids which couple sequentially to each last-coupled amino acid residue to create a peptide containing at least one Asp or Glu residue the side chain of which is protected and simultaneously cleaving said peptide from said resin and deprotecting said side chain in a manner that the C-terminus of said peptide is --CONHQ.
Claim 19
Independent Claim
163 words
A method for synthesizing a peptide containing at least one Asp or Glu residue within its chain and having a substituted amide at its C-terminus using solid phase synthesis, which method comprises providing a synthetic resin having a plurality of substituted p-benzl moieties of the formula --C 6 H 4 CH 2 NHQ wherein Q is lower alkyl, lower fluoroalkyl, phenyl or substituted phenyl, reacting said synthetic resin with an amino acid having its alpha-amino group blocked to cause coupling of the carboxyl group thereof to the nitrogen of said moiety by a substituted amide linkage, deblocking said alpha-amino group, repeating said reacting and deblocking steps with such additional amino acids which couple sequentially to each last-coupled amino acid residue to create a peptide containing at least one Asp or Glu residue the side chain of which is protected and simultaneously cleaving said peptide from said resin and deprotecting said side chain in a manner that the C-terminus of said peptide is --CONHQ.
Claim 20
Dependent Claim
25 words
The method in accordance with claim 17 wherein said resin is a cross-linked polystyrene resin and said cleaving and deprotecting is carried out with HF.
Claim 21
Dependent Claim
22 words
A method in accordance with claim 17 wherein the side chain of the Asp or Glu residue is protected with benzyl ester.
Claim 22
Independent Claim
135 words
A method for synthesizing a peptide having an N-nitroanilide at its C-terminus using solid phase synthesis, which method comprises providing a synthetic resin having a plurality of moieties of the formula --C 6 H 4 CH 2 NHC 6 H 4 NH 2 , reacting said synthetic resin with an amino acid having its alpha-amino group blocked so as to cause coupling of the carboxyl group thereof to the amino group of said moiety by an amide linkage, deblocking said alpha-amino group, repeating said reacting and deblocking steps with such additional amino acids which couple sequentially to each last-coupled amino acid residue to create a peptide of desired length, cleaving said peptide from said resin in a manner such that the C-terminus of said peptide is an N-aminoanilide and oxidizing said N-aminoanilide to an N-nitroanilide.
Claim 23
Independent Claim
135 words
A method for synthesizing a peptide having an N-nitroanilide at its C-terminus using solid phase synthesis, which method comprises providing a synthetic resin having a plurality of moieties of the formula --C 6 H 4 CH 2 NHC 6 H 4 NH 2 , reacting said synthetic resin with an amino acid having its alpha-amino group blocked so as to cause coupling of the carboxyl group thereof to the amino group of said moiety by an amide linkage, deblocking said alpha-amino group, repeating said reacting and deblocking steps with such additional amino acids which couple sequentially to each last-coupled amino acid residue to create a peptide of desired length, cleaving said peptide from said resin in a manner such that the C-terminus of said peptide is an N-aminoanilide and oxidizing said N-aminoanilide to an N-nitroanilide.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1989-02-10 | 1989-08-12 | 1990-02-11 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-02-10 | 1993-08-12 | 1994-02-11 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-02-10 | 1997-08-12 | 1998-02-11 | $7700.00 | Window Closed / Lapsed |