Proteinic active substances

Claim 1

A protein fraction which is selected from the group consisting of KS-I, KS-II and KS-III which is prepared by the process of culturing a pathogenic strain of Bordetella pertussis, isolating from the cultured cells or culture medium a protein which has a molecular weight of 77,000±6,400 as determined by gel filtration and has insulin secretion promoting activity for a mammal, which protein in 0.1 M phosphate buffer (pH 7.0) is adsorbed on a hydroxyapatite column and eluted therefrom with 0.1 M phosphate buffer of pH 7.0 containing 0.5 M NaCl and which protein in SDS-polyacrylamide-gel electrophoresis, wherein an incubated mixture of 1% SDS, 1% mercaptoethanol and 4 M urea with 50 μg/tube of said protein is applied to 10% polyacrylamide-gel containing 1% SDS in 4-hour current application of 8 mA/gel, gives three main peptide-bands, mixing said protein into a solution of protein denaturant to dissociate it and recovering from the resultant mixture said protein fraction characterized by the following properties: KS--I molecular weight: 63,000±5,200 as determined by gel filtration; chemical composition: protein content as determined by the Lowry's method being ocer 97% by weight, the glucide content by the phenol-H 2 SO 4 method being about 1% by weight and the lipid content being lower than the limit of detection, the amino acids composition of the protein moiety: asparatic acid 8.1%; threonine 8.5%; serine 7.9%; glutamic acid 9.6%; proline 5.0%; glycine 10.0%; alanine 9.5%; cystine/2 1.1%; valine 6.2%; methionine 1.8%; isoleucine 4.5%; leucine 6.5%; tyrosine 7.6%; phenylalanine 2.8%; lysine 2.1%; histidine 1.6% and arginine 7.3%; isoelectric pH value of 5.6±0.3, and disc electrophoretic pattern, using acrylamide (polyacrylamide concentration, 7.5%; 1 N KOH-glacial acetic acid buffer (pH 4.3)) disc electrophoresis, giving a very sharp single band on the anode side; KS--II molecular weight: 31,000±4,500 as determined by gel filtration; chemical composition: protein content as determined by the Lowry method being over 95% by weight, the glucide content by the phenol-H 2 SO 4 method being about 1% by weight and the lipid content being lower than the limit of detection; the amino acid composition of the protein moiety: asparatic acid 9.3%; threonine 7.7%; serine 9.8%; glutamic acid 11.8%; proline 4.6%; glycine 9.8%; alanine 11.3%; cystine/2 N.D.; valine 7.4%; methionine 1.9%; isoleucine 4.0%; leucine 5.3%; tyrosine 6.2%; phenylalanine 2.5%; lysine 1.9%; histidine 1.1%; and arginine 5.4%; isoelectric pH value of 5.4±0.4 and disc electrophoretic pattern, using acrylamide (polyacrylamide concentration 7.5%; 1 N KOH-glacial acetic acid buffer (pH 4.3)) disc electrophoresis, giving a very sharp single band on the anode side; KS--III molecular weight: 12,000±1,500 as determined by gel filtration, chemical composition: protein content as determined by the Lowry method being over 96% by weight the glucide content by the phenol-H 2 SO 4 method being about 1% by weight and the lipid content being lower than the limit of detection, the amino acids composition of the protein moiety: asparatic acid 5.3; threonine 4.8%; serine 6.2%; glutamic acid 9.6%; proline 9.3%; glycine 8.1%; alanine 9.3%; cystine/2 2.3%; valine 10.2%; methionine 6.0%; isoleucine 2.2%; leucine 8.6%; tyrosine 2.5%; phenylalanine 4.5%; lysine 5.8%; histidine 0.5%; and arginine 4.8%; isoelectric pH value of 8.3±0.3, and disc electrophoretic pattern using acrylamide (polyacrylamide concentration 7.5%; 1 N KOH-glacial acetic acid buffer (pH 4.3)) disc electrophoresis, giving a very sharp single band on the cathode side.

Claim 12

A method of treating a patient suffering from diabetes which comprises administering to said patient an amount of a protein fraction defined by any of claims 1 to 5 capable of promoting insulin secretion in said patient.