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US5066716A
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Synthesis of chloroacetyl and bromoacetyl modified peptides for the preparation of synthetic peptide polymers, conjugated peptides, and cyclic peptides
Assignee: United States Department of Health and Human Services
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Filing: 1988-12-13
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Granted: 1991-11-19
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Expires: 2008-12-13
Abstract
Abstract A method to incorporate bromoacetyl and chloroacetyl moieties on amino groups of synthetic peptides using a standard program with an automated peptide synthesizer has been developed. The bromoacetyl and chloroacetyl-derivatized peptides react well with sulfhydryl-containing proteins and with peptides containing cysteine residues. Autopolymerization or cyclization occurs by reaction of the free sulfhydryl of cysteine in a peptide with the bromoacetyl group (or chloroacetyl group) and reactions can generally be controlled by controlling the concentrations of starting peptide in neutral pH buffers. Analytical methods for evaluating the polymers or cyclized peptides include gel filtration chromatography, reverse phase HPLC, SDS-PAGE and amino acid analysis where the degree of reaction can be evaluated by quantifying the amount of S-carboxymethylcysteine formed after HCl hydrolysis. N-bromoacetyl-derivatized peptides are useful as reagents for potential peptide immunogens, vaccines and therapeutics, and for substances such as peptides linked to polymers, plastics, enamels, and ceramics.
Inventors
- 👤 Frank A. Robey
- 👤 Raymond L. Fields
- 👤 Wolfgang Lindner
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 02:39
CPC Classifications
C; C07; C07K; C07K1/00; C07K1/107; C07K1/1072; C07K1/1077
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
63 words
A method for the preparation of bromoacetyl or chloroacetyl derivatized peptides, which comprises the steps of: coupling a bromoacetyl or chloroacetyl group onto a fully protected peptide to form an amide linkage between said bromoacetyl or chloroacetyl group and said fully protected peptide; and deprotecting said fully protected peptide while still preserving the presence of said bromoacetyl or chloroacetyl group on said peptide.
Claim 2
Independent Claim
63 words
A method for the preparation of bromoacetyl or chloroacetyl derivatized peptides, which comprises the steps of: coupling a bromoacetyl or chloroacetyl group onto a fully protected peptide to form an amide linkage between said bromoacetyl or chloroacetyl group and said fully protected peptide; and deprotecting said fully protected peptide while still preserving the presence of said bromoacetyl or chloroacetyl group on said peptide.
Claim 3
Dependent Claim
15 words
The method according to claim 1, wherein said derivatized peptide is an N-bromoacetyl derivatized peptide.
Claim 4
Dependent Claim
15 words
The method according to claim 1, wherein said derivatized peptide is an N-chloroacetyl derivatized peptide.
Claim 5
Dependent Claim
28 words
The method according to claim 1, wherein said fully protected peptide contains cysteine, a group with an SH group, or a reactive amino group in the peptide chain.
Claim 6
Dependent Claim
47 words
The method according to claim 1, which comprises forming a symmetric anhydride of bromoacetic acid or chloroacetic acid; reacting said bromoacetic anhydride or chloroacetic acid anhydride with an N-terminus of a protected peptide to form said amide linkage; and deprotecting said peptide by treatment with an acid.
Claim 7
Dependent Claim
28 words
The method according to claim 5, wherein the formation of said symmetric anhydride of bromoacetic acid or chloroacetic acid occurs at about 0° C. to about 30° C.
Claim 8
Dependent Claim
33 words
The method according to claim 5, wherein the reaction of the bromoacetic acid and anhydride or chloroacetic acid anhydride with the protected peptide is conducted at about 0° C. to about 30° C.
Claim 9
Dependent Claim
22 words
The method according to claim 5, wherein the treatment with the acid is conducted at about -5° C. to about 5° C.
Claim 10
Dependent Claim
22 words
The method according to claim 8, wherein the treatment with the acid is conducted for about 10 minutes to about 3 hours.
Claim 11
Dependent Claim
35 words
A method according to claim 4, which further comprises polymerizing or cyclizing said bromoacetyl or chloroacetyl derivatized peptide by treating said derivatized peptide in a solution having a pH of about 5 to about 11.
Claim 12
Dependent Claim
27 words
The method according to claim 10, wherein said polymerization or cyclization occurs in a solution of NaHCO 3 or NaHPO 4 having a pH of about 8.
Claim 13
Dependent Claim
17 words
The method according to claim 11, which is conducted at about 15° C. to about 30° C.
Claim 14
Dependent Claim
16 words
The method according to claim 12, which is conducted for about 0.5 to about 48 hours.
Claim 15
Dependent Claim
36 words
The method according to claim 1, which further comprises reacting the bromoacetyl or chloroacetyl derivatized peptide with a carrier protein containing cysteine, a group having an SH group, or a reactive amine to form a conjugate.
Claim 16
Dependent Claim
30 words
The method according to claim 14, wherein the reaction is conducted for about 0.5 hours to about 48 hours at a temperature of about 15° C. to about 30° C.
Claim 17
Dependent Claim
29 words
The method according to claim 14, wherein said carrier protein is a member selected from the group consisting of bovine serum albumin, chicken serum albumin, and keyhole lymphet hemocyanin.
Claim 18
Dependent Claim
23 words
A bromoacetyl or chloroacetyl derivatized peptide in which the bromoacetyl group or chloroacetyl group is bonded to the peptide via an amide linkage.
Claim 19
Dependent Claim
23 words
A bromoacetyl or chloroacetyl derivatized peptide in which the bromoacetyl group or chloroacetyl group is bonded to the peptide via an amide linkage.
Claim 20
Dependent Claim
9 words
A polymer of the derivatized peptide of claim 17.
Claim 21
Dependent Claim
10 words
A cyclized form of the derivatized peptide of claim 18.
Claim 22
Dependent Claim
13 words
A conjugate of a carrier protein and the derivatized peptide of claim 17.
Claim 23
Independent Claim
114 words
A method for the preparation of bromoacetyl or chloroacetyl derivatized proteins, which comprises: coupling a bromoacetyl or chloroacetyl group onto a fully protected peptide at a pH of about 5 to about 11 for about 10 minutes to about 48 hours at about -5° C. to 30° C. in an aprotic polar solvent to form an amide linkage between said bromoacetyl or chloroacetyl group and said fully protected peptide, and deprotecting said fully protected peptide while still preserving the presence of said bromoacetyl or chloroacetyl group on said peptide at a temperature of about -5° C. to about 5° C. for about 10 minutes to about 3 hours in trifluoroacetic acid or hydrogen fluoride.
Claim 24
Independent Claim
114 words
A method for the preparation of bromoacetyl or chloroacetyl derivatized proteins, which comprises: coupling a bromoacetyl or chloroacetyl group onto a fully protected peptide at a pH of about 5 to about 11 for about 10 minutes to about 48 hours at about -5° C. to 30° C. in an aprotic polar solvent to form an amide linkage between said bromoacetyl or chloroacetyl group and said fully protected peptide, and deprotecting said fully protected peptide while still preserving the presence of said bromoacetyl or chloroacetyl group on said peptide at a temperature of about -5° C. to about 5° C. for about 10 minutes to about 3 hours in trifluoroacetic acid or hydrogen fluoride.
Claim 25
Dependent Claim
30 words
A peptide carrier conjugate produced by a process which comprises reacting the derivatized peptide of claim 17 with a carrier selected from the group consisting of polymers, enamels and ceramics.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1994-11-18 | 1995-05-20 | 1995-11-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-11-18 | 1999-05-20 | 1999-11-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-11-18 | 2003-05-20 | 2003-11-19 | $7700.00 | Window Closed / Lapsed |