Vasoactive lung polypeptides

Claim 1

A method for the extraction and isolation of vasoactive peptides from the lungs of animals, one of said peptides being the fraction of hatched area B of FIG. 2, and the other of said peptides being the fraction of hatched area A of FIG. 2 and exhibiting the property of relaxing isolated non-vascular, smooth muscle organs, said method comprising the steps of collecting, boiling and mincing lung portions, extracting the resulting lung portions with acetic acid at a temperature of about 0°-5° C., filtering the resulting extract, and subjecting the extract to alginic acid absorption by the steps comprising lowering the pH with HCl acid and stirring with alginic acid, allowing the alginic acid mixture to settle and discarding supernatant liquid, recovering the resulting sediment, washing with dilute HCl and eluting from the alginic acid with HCl, adding sodium chloride to the eluate and collecting a salt precipitate, dissolving the salt precipitate in water, neutralizing the resulting solution to a pH of about 7, heating the solution with vigorous stirring and then extracting it with an alcohol, acidifying with mineral acid and collecting an alcoholic extract material, subjecting the alcoholic extract material to gel chromatography over a cross-linked dextran at least one time while dissolved in an organic acid, collecting fractions from the beginning of introduction of the extract into the chromatographic column, combining the recovered fractions, saturating with sodium chloride and salting out the product, collecting the fractions from the gel chromatography in two parts, testing for the most active fractions by tests of femoral blood flow on anesthetized dogs and on isolated smooth muscle organs, subjecting the active fractions to at least one ion-exchange chromatography over carboxymethyl cellulose while dissolved in a buffer solution at a pH of 6-7, centrifuging to eliminate insoluble materials, collecting separate fractions and treating with alginic acid for absorption of peptides and eluting with mineral acid, freeze drying and lyophilizing the resulting fractions, and identifying the separate active fractions by characterization for effect on blood flow and smooth muscle organs; said active fractions being said fraction of hatched area B and said fraction of hatched area A.

Claim 2

A method for the extraction and isolation of vasoactive peptides from the lungs of animals, one of said peptides being the fraction of hatched area B of FIG. 2, and the other of said peptides being the fraction of hatched area A of FIG. 2 and exhibiting the property of relaxing isolated non-vascular, smooth muscle organs, said method comprising the steps of collecting, boiling and mincing lung portions, extracting the resulting lung portions with acetic acid at a temperature of about 0°-5° C., filtering the resulting extract, and subjecting the extract to alginic acid absorption by the steps comprising lowering the pH with HCl acid and stirring with alginic acid, allowing the alginic acid mixture to settle and discarding supernatant liquid, recovering the resulting sediment, washing with dilute HCl and eluting from the alginic acid with HCl, adding sodium chloride to the eluate and collecting a salt precipitate, dissolving the salt precipitate in water, neutralizing the resulting solution to a pH of about 7, heating the solution with vigorous stirring and then extracting it with an alcohol, acidifying with mineral acid and collecting an alcoholic extract material, subjecting the alcoholic extract material to gel chromatography over a cross-linked dextran at least one time while dissolved in an organic acid, collecting fractions from the beginning of introduction of the extract into the chromatographic column, combining the recovered fractions, saturating with sodium chloride and salting out the product, collecting the fractions from the gel chromatography in two parts, testing for the most active fractions by tests of femoral blood flow on anesthetized dogs and on isolated smooth muscle organs, subjecting the active fractions to at least one ion-exchange chromatography over carboxymethyl cellulose while dissolved in a buffer solution at a pH of 6-7, centrifuging to eliminate insoluble materials, collecting separate fractions and treating with alginic acid for absorption of peptides and eluting with mineral acid, freeze drying and lyophilizing the resulting fractions, and identifying the separate active fractions by characterization for effect on blood flow and smooth muscle organs; said active fractions being said fraction of hatched area B and said fraction of hatched area A.