US3879371A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Isolation of vasoactive intestinal peptide
Assignee: Research Corp
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Filing: 1971-09-17
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Granted: 1975-04-22
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Expires: 1991-09-17
Abstract
Abstract A polypeptide having biological action including systemic vasodilation, hypotension, increased cardiac output, respiratory stimulation, and hyperglycemia isolated from intestines of animals such as a hog. The isolation procedure for purification includes two steps of ion-exchange chromatography, gel chromatography and countercurrent distribution separation. The polypeptide, VIP, has the amino acid structure: His (Ala, Asp, Ser, Val). Phe, Thr. (Asx2). Tyr. Thr. Arg. Leu. Arg. (Lys, Glx). Met. Ala (Val, Lys, Lys). Tyr. Leu. Asn. Ser. Ile. Leu. Asn. NH2. Therapeutic biological actions may be produced in animals and humans by intravenous injection by .02-10 Mu g VIP per kg of body weight.
Inventors
- 👤 Sami I Said
- 👤 Viktor Mutt
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 15:49
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: 8
Claim 1
Independent Claim
137 words
A METHOD OF EXTRACTING AND ISOLATING VIP, A NEW BIOLOGICALLY ACTIVE POLYPEPTIDE FROM THE INTESTINES OF MAMMALS COMPRISING: EXTRACTING CRUDE VIP AND SECRETIN FROM SAID INTESTINES, SUBJECTING SAID CRUDE VIP AND SECRETIN TO A FIRST IONEXCHANGE CHROMATOGRAPHY, COLLECTING FRACTIONS CONTAINING CURDE VIP AFTER ALL THE SECRETIN HAS BEEN REMOVED, SUBJECTING SAID CRUDE VIP FURTHER TO GEL CHROMATOGRAPHY, COLLECTING FRACTIONS HAVING APPROXIMATELY THE GREATEST OPTICAL DENSITY MEASURED AT 280 M$ CONTAINING FURTHER PURIFIED VIP, SUBJECTING THE SO PURIFIED VIP TO A SECOND ION-EXCHANGE CHROMATOGRAPHY AND COLLECTING FRACTIONS AFTER THE OPTICAL DENSITY MEASURED AT 280 M$ OF SUCCESSIVE FRACTIONS FALLS BELOW APPROXIMATELY ONE-FOURTH OF THE PEAK VALUE, AND THEREAFTER, SUBJECTING THE FURTHER PURIFIED VIP TO COUNTERCURRENT DISTRIBUTION SEPARATION AND COLLECTING PURIFIED VIP AT APPROXIMATELY THOSE FRACTIONS HAVING THE FIRST FULL PEAK VALUE OF OPTICAL DENSITY MEASURED AT 215 M$.
Claim 2
Independent Claim
137 words
A METHOD OF EXTRACTING AND ISOLATING VIP, A NEW BIOLOGICALLY ACTIVE POLYPEPTIDE FROM THE INTESTINES OF MAMMALS COMPRISING: EXTRACTING CRUDE VIP AND SECRETIN FROM SAID INTESTINES, SUBJECTING SAID CRUDE VIP AND SECRETIN TO A FIRST IONEXCHANGE CHROMATOGRAPHY, COLLECTING FRACTIONS CONTAINING CURDE VIP AFTER ALL THE SECRETIN HAS BEEN REMOVED, SUBJECTING SAID CRUDE VIP FURTHER TO GEL CHROMATOGRAPHY, COLLECTING FRACTIONS HAVING APPROXIMATELY THE GREATEST OPTICAL DENSITY MEASURED AT 280 M$ CONTAINING FURTHER PURIFIED VIP, SUBJECTING THE SO PURIFIED VIP TO A SECOND ION-EXCHANGE CHROMATOGRAPHY AND COLLECTING FRACTIONS AFTER THE OPTICAL DENSITY MEASURED AT 280 M$ OF SUCCESSIVE FRACTIONS FALLS BELOW APPROXIMATELY ONE-FOURTH OF THE PEAK VALUE, AND THEREAFTER, SUBJECTING THE FURTHER PURIFIED VIP TO COUNTERCURRENT DISTRIBUTION SEPARATION AND COLLECTING PURIFIED VIP AT APPROXIMATELY THOSE FRACTIONS HAVING THE FIRST FULL PEAK VALUE OF OPTICAL DENSITY MEASURED AT 215 M$.
Claim 3
Independent Claim
141 words
A method of extracting and isolating VIP, a new biologically active polypeptide from the intestines of mammals comprising: extracting crude VIP and secretin from said intestines, subjecting said crude VIP and secretin to a first ion-exchange chromatography, collecting fractions containing crude VIP after all the secretin has been removed, subjecting said crude VIP further to gel chromatography, collecting fractions having approximately the greatest optical density measured at 280 m Mu containing further purified VIP, subjecting the so purified VIP to a second ion-exchange chromatography and collecting fractions after the optical density measured at 280 m Mu of successive fractions falls below approximately one-fourth of the peak value, and thereafter, subjecting the further purified VIP to countercurrent distribution separation and collecting purified VIP at approximately those fractions having the first full peak value of optical density measured at 215 m Mu .
Claim 4
Independent Claim
141 words
A method of extracting and isolating VIP, a new biologically active polypeptide from the intestines of mammals comprising: extracting crude VIP and secretin from said intestines, subjecting said crude VIP and secretin to a first ion-exchange chromatography, collecting fractions containing crude VIP after all the secretin has been removed, subjecting said crude VIP further to gel chromatography, collecting fractions having approximately the greatest optical density measured at 280 m Mu containing further purified VIP, subjecting the so purified VIP to a second ion-exchange chromatography and collecting fractions after the optical density measured at 280 m Mu of successive fractions falls below approximately one-fourth of the peak value, and thereafter, subjecting the further purified VIP to countercurrent distribution separation and collecting purified VIP at approximately those fractions having the first full peak value of optical density measured at 215 m Mu .
Claim 5
Dependent Claim
36 words
The method of claim 1 including dissolving said crude VIP in a buffer at approximately pH 7-9 for said first and second ion-exchange chromatography, dissolving said crude VIP in an organic acid in said gel chromatography.
Claim 6
Dependent Claim
71 words
The method of claim 1 including dissolving said crude VIP in a buffer selected from sodium and ammonium carbonates, bicarbonates, and phosphates and acid phosphates at a pH of 7-9 approximately, prior to the first ion exchange chromatography, dissolving said crude VIP for said gel chromatography in an organic acid, said gel filter being selected from a cross-linked dextran and cellulose, using an alcohol-buffer, as defined above, system to isolate VIP.
Claim 7
Dependent Claim
52 words
The method of claim 3 including said buffers being Na2HPO4 for said first ion exchange, NH4HCO3 for said second ion exchange, said organic acid with said gel chromatography being acetic acid and said countercurrent distribution separation system being 1-butanol and NH4HCO3, and said first and second ion-exchange chromatography filter material being CMC.
Claim 8
Dependent Claim
35 words
The method of claim 1 including eluting the crude VIP with acid to obtain the fractions with VIP in the first ion-exchange chromatography, converting said VIP to an acetate and freeze-drying said crude VIP product.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1978-04-21 | 1978-10-21 | 1979-04-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1982-04-21 | 1982-10-21 | 1983-04-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1986-04-21 | 1986-10-21 | 1987-04-22 | $7700.00 | Window Closed / Lapsed |