US4430266A
Expired / Lapsed
US
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Process for producing an insulin precursor
Assignee: Eli Lilly and Co
•
Filing: 1982-02-16
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Granted: 1984-02-07
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Expires: 2002-02-16
Abstract
Abstract A proinsulin-like disulfide insulin precursor is produced from its corresponding linear chain S-sulfonate insulin precursor by reacting the S-sulfonate with a mercaptan in an amount which provides from about 1 to about 5 --SH moieties per --SSO 3 - moiety in an aqueous medium at a pH of from about 7 to about 11.5 and at an S-sulfonate concentration of up to about 10 mg. per ml. of aqueous medium.
Inventors
- 👤 Bruce H. Frank
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 02:49
CPC Classifications
C; C07; C07K; C07K1/00; C07K1/06; C07K1/061; C07K1/067; C07K1/12; C07K14/00; C07K14/435; C07K14/575; C07K14/62; Y; Y10; Y10S; Y10S930/00; Y10S930/01; Y10S930/27
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
196 words
A process for the production of an insulin precursor of the formula ##STR4## in which R is hydrogen, a chemically or enzymatically cleavable amino acid residue, or a chemically or enzymatically cleavable peptide moiety having at least two amino acid residues, Y is ##STR5## in which Z is Ala, Thr, or Ser; the moiety from A-1 to A-21 is an insulin A-chain; the moiety from B-1 to B-30 is an insulin B-chain; and X is a moiety which is joined to the insulin A-chain at the amino group of A-1 and to the insulin B-chain at the ε-amino group of B-29 or the carboxyl group of B-30, which moiety can be enzymatically or chemically cleaved from and without disruption of both the A-chain and the B-chain, which comprises reacting an S-sulfonate of the formula ##STR6## in which R, X, and Y are as aforedefined, with a mercaptan in an amount which provides from about 1 to about 5 --SH moieties per each --SSO 3 - moiety in an aqueous medium at a pH of from about 7 to about 11.5 and at an S-sulfonate concentration of up to about 10 mg. per ml. of aqueous medium.
Claim 2
Dependent Claim
196 words
A process for the production of an insulin precursor of the formula ##STR4## in which R is hydrogen, a chemically or enzymatically cleavable amino acid residue, or a chemically or enzymatically cleavable peptide moiety having at least two amino acid residues, Y is ##STR5## in which Z is Ala, Thr, or Ser; the moiety from A-1 to A-21 is an insulin A-chain; the moiety from B-1 to B-30 is an insulin B-chain; and X is a moiety which is joined to the insulin A-chain at the amino group of A-1 and to the insulin B-chain at the ε-amino group of B-29 or the carboxyl group of B-30, which moiety can be enzymatically or chemically cleaved from and without disruption of both the A-chain and the B-chain, which comprises reacting an S-sulfonate of the formula ##STR6## in which R, X, and Y are as aforedefined, with a mercaptan in an amount which provides from about 1 to about 5 --SH moieties per each --SSO 3 - moiety in an aqueous medium at a pH of from about 7 to about 11.5 and at an S-sulfonate concentration of up to about 10 mg. per ml. of aqueous medium.
Claim 3
Dependent Claim
9 words
Process of claim 1, in which R is hydrogen.
Claim 4
Dependent Claim
22 words
Process of claim 2, in which X is a peptide moiety joined to the insulin B-chain at the carboxyl group of B-30.
Claim 5
Dependent Claim
19 words
Process of claim 3, in which X is -Arg-X'-Arg- in which X' is at least one amino acid residue.
Claim 6
Dependent Claim
22 words
Process of claim 4, in which X is -Arg-Arg-X 2 -Lys-Arg- in which X 2 is at least one amino acid residue.
Claim 7
Dependent Claim
21 words
Process of claim 5, in which the reaction is carried out at a pH of from about 9.5 to about 10.5.
Claim 8
Dependent Claim
28 words
Process of claim 6, in which the mercaptan is present in an amount which provides from about 2 to about 4 --SH moieties per --SSO 3 - moiety.
Claim 9
Dependent Claim
19 words
Process of claim 7, in which the reaction is carried out in the substantial absence of an oxidizing agent.
Claim 10
Dependent Claim
24 words
Process of claim 8, in which the concentration of S-sulfonate is from about 0.05 milligrams to about 2 milligrams per milliliter of aqueous medium.
Claim 11
Dependent Claim
10 words
Process of claim 9, in which the mercaptan is 2-mercaptoethanol.
Claim 12
Dependent Claim
23 words
Process of claim 10, in which the reaction is carried out at a temperature of from about 2° C. to about 8° C.
Claim 13
Dependent Claim
23 words
Process of claim 11, in which the reaction is carried out at a temperature of from about 4° C. to about 6° C.
Claim 14
Dependent Claim
39 words
Process of claim 12, in which the reaction mixture is prepared at about room temperature, and the reaction is allowed to proceed with cooling to a temperature within the range of from about 4° C. to about 6° C.
Claim 15
Dependent Claim
32 words
Process of claim 13, in which the pH of the reaction mixture is maintained by addition of a buffering agent at a concentration of from about 0.01 N to about 0.5 N.
Claim 16
Dependent Claim
11 words
Process of claim 14, in which the buffering agent is glycine.
Claim 17
Dependent Claim
22 words
Process of claim 15, in which the insulin A-chain and the insulin B-chain of the S-sulfonate have the structure of human insulin.
Claim 18
Dependent Claim
14 words
Process of claim 16, in which X is the connecting peptide of human insulin.
References Cited (Backward Citations)
US3847893A
Cited
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) | 1987-02-06 | 1987-08-08 | 1988-02-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-02-06 | 1991-08-08 | 1992-02-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-02-06 | 1995-08-08 | 1996-02-07 | $7700.00 | Window Closed / Lapsed |