Patent US4016258A successfully refreshed from Google Patents.
US4016258A
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Vasoactive intestinal peptide from fowl
Assignee: Research Corp
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Filing: 1975-08-05
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Granted: 1977-04-05
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Expires: 1995-08-05
Abstract
Abstract A polypeptide having important biological activity including systemic vasodilation, hypotension, increased cardiac output and bronchodilation is isolated from intestines of chickens. The isolation procedure for purification includes extraction, ion-exchange chromatography, gel chromatography and countercurrent distribution separation. The polypeptide has the general amino acid structure: His-Ser-Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr-Ser-Arg-Phe-Arg-Lys-Gln-Met-Ala-Val-Lys-Lys-Tyr-Leu-Asn-Ser-Val-Leu-Thr-NH 2 . Therapeutic biological actions may be produced in animals and humans by administration of 0.02-10 μg of the peptide per kg of body weight.
Inventors
- 👤 Sami I. Said
- 👤 Viktor Mutt
- 👤 Ann Nilsson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 16:01
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/575; C07K14/57563; A; A61; A61K; A61K38/00; Y; Y10; Y10S; Y10S930/00; Y10S930/01; Y10S930/17
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
102 words
A method for the extraction and isolation of a vasoactive peptide from the intestines of fowl having the following general structure: His-Ser-Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr-Ser-Arg-Phe-Arg-Lys-Gln-Met-Ala-Val-Lys-Lys-Tyr-Leu-Asn-Ser-Val-Leu-Thr-NH 2 which comprises extracting crude polypeptide concentrate from said intestines, purifying said crude concentrate at least by absorption with alginic acid, collecting fractions containing said crude concentrate, subjecting said crude concentrate to gel chromatography, collecting fractions thereof, subjecting the resultant fractions to extraction with a lower alkyl alcohol, recovering the product, subjecting the extracted product to ion-exchange chromatography, collecting fractions of the active product, and thereafter subjecting the further purified polypeptide to countercurrent distribution separation, and collecting the purified polypeptide.
Claim 2
Independent Claim
102 words
A method for the extraction and isolation of a vasoactive peptide from the intestines of fowl having the following general structure: His-Ser-Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr-Ser-Arg-Phe-Arg-Lys-Gln-Met-Ala-Val-Lys-Lys-Tyr-Leu-Asn-Ser-Val-Leu-Thr-NH 2 which comprises extracting crude polypeptide concentrate from said intestines, purifying said crude concentrate at least by absorption with alginic acid, collecting fractions containing said crude concentrate, subjecting said crude concentrate to gel chromatography, collecting fractions thereof, subjecting the resultant fractions to extraction with a lower alkyl alcohol, recovering the product, subjecting the extracted product to ion-exchange chromatography, collecting fractions of the active product, and thereafter subjecting the further purified polypeptide to countercurrent distribution separation, and collecting the purified polypeptide.
Claim 3
Dependent Claim
34 words
The method of claim 1 including dissolving said crude peptide in a buffer at approximately pH 6-7 for said ion-exchange chromatography, and dissolving said crude peptide in an organic acid in said gel chromatography.
Claim 4
Dependent Claim
60 words
The method of claim 2 including dissolving said crude peptide in a buffer selected from sodium and ammonium carbonates, bicarbonates and phosphates and acid phosphates at a pH of 6.2-6.6 approximately, dissolving said crude peptide for said gel chromatography in an organic acid, said gel filter being selected from Sephadex and cellulose, using an alcohol-buffer, system to isolate said peptide.
Claim 5
Dependent Claim
48 words
The method of claim 3 including said buffers being Na 2 HPO 4 for said ion-exchange, said organic acid with said gel chromatography being acetic acid and said countercurrent distribution separation system being 1-butanol and NH 4 HCO 3 , and said ion-exchange chromatography filter material being CMC.
Claim 6
Dependent Claim
23 words
The method of claim 1 including eluting the crude peptide with acid to obtain the fractions with peptide in the ion-exchange chromatographic step.
Claim 7
Dependent Claim
15 words
The method of claim 4 including maintaining the pH at 6.4 in said ion-exchange chromatography.
Claim 8
Dependent Claim
11 words
The novel purified polypeptide having the following structure: His-Ser-Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr-Ser-Arg-Phe-Arg-Lys-Gln-Met-Ala-Val-Lys-Lys-Tyr-Leu-Asn-Ser-Val-Leu-Thr-NH 2 .
Claim 9
Dependent Claim
11 words
The novel purified polypeptide having the following structure: His-Ser-Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr-Ser-Arg-Phe-Arg-Lys-Gln-Met-Ala-Val-Lys-Lys-Tyr-Leu-Asn-Ser-Val-Leu-Thr-NH 2 .
Claim 10
Dependent Claim
19 words
A therapeutic composition comprising a therapeutically effective amount of the polypeptide of claim 7 and a physiologically acceptable carrier.
Claim 11
Dependent Claim
21 words
A composition according to claim 8 wherein the composition contains about 0.001 to 0.1 weight percent of polypeptide in the carrier.
Claim 12
Dependent Claim
24 words
A method for producing bronchodilation in animals and humans comprising administration by inhalation of a therapeutically effective amount of the polypeptide of claim 7.
References Cited (Backward Citations)
US3879371A
Cited
US3880826A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1980-04-04 | 1980-10-04 | 1981-04-05 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1984-04-04 | 1984-10-04 | 1985-04-05 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1988-04-04 | 1988-10-04 | 1989-04-05 | $7700.00 | Window Closed / Lapsed |