Patent GB-874230-A (GB)
Title: Cobalaminí¬peptide complexes
Inventor: HEATHCOTE JOHN GODFREY
IPC: C12P19/42
Filed: 1957-04-04
Abstract: The invention comprises a cobalaminpeptide complex which is effective on oral administration against pernicious anaemia, resulting from the degrading of a cobalamin-containing product of microbial fermentation and a process for producing said complex wherein a suitable nutrient medium is fermented with a cobalamin-producing organism and thereafter subjecting a product p  of the fermentation containing 'bond' cobalamin, to a degradation treatment that results in the production of at least one cobalamin-peptide complex having a molecular weight below about 15,000 and recovering the complex in a non-toxic form which is effective against pernicious anaemia on oral administration.  As cobalamin-producing organism there may be used a member of the genus Propionibacterium, e.g. P. freudenreichii. Degradation may take the form of treatment with cyanide ions, e.g. with potassium cyanide treatment with a proteolytic enzyme, e.g. pepsin, trypsin, chrymotrypsin or papain, by maintaining it at an acid pH, e.g. in the range 1-3, or by heating the cobalamin-containing product of the microbial fermentation.  Prior to the degradation step the liquid material resulting from fermentation may be concentrated, e.g. by adsorption from solution on to a suitable ion exchange resin and by solvent extraction to produce an aqueous solution containing a concentration of total cobalamins exceeding 500 micrograms per millilitre.  Subsequent to the degradation, the aqueous solution containing the complex may be further concentrated or treated with a precipitant, e.g. ammonium sulphate to obtain the cobalamin-peptide complex in solid form.  The cobalamin-peptide complex may be further characterised in that it results from degrading a cobalamin-containing product of microbial fermentation until it dialyses through a regenerated cellulose membrane having an average pore size of about 24 Angstrom units against water, and that the peptide portion of the complex on acid hydrolysis yields at least one of the following acids: glutamic and aspartic acids, glycine, valine, proline, arginine, cysteine or cystine, serine alanine, leucine, isoleucine, phenylalanine, lysine, histidine and threonine; and that the peptide portion of the complex on alkaline hydrolysis yields tryptophane, and by its absorption spectra.  Specification 665,485 is referred to.ALSO:A pharmaceutical preparation comprises a cobalamin-peptide complex which is effective on oral administration against pernicious anaemia, resulting from the degrading of a cobalamin-containing product of microbial fermentation, but which is substantially free from "uncombined" cobalamin and contains "bound" cobalamin at most in minor amount compared with the amount of complex, diluted with a non-toxic material, e.g. water, liquid or mannitol.  The preparations may be put into tablet or capsule form or may be employed in liquid form by dissolving or suspending in water or other liquids containing buffering agents. The palatability of such preparations may be increased by the addition of flavouring or sweetening agents. Specification 665,485 is referred to.