# US 4,397,840 A · Novel erythropoietin product and method for the preparation thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 4,397,840 A - Document ID: US4397840A - Status: Granted - Kind: A - Application number: 06/354,302 - Filed: 1982-03-03 - Granted: 1983-08-09 - Earliest claimed: 1981-03-11 - Assignees: Ajinomoto Co., Inc.; Japan Chemical Research Co., Ltd. - Inventors: Kenji Takezawa; Hajima Hiratani - Web page: https://patentgrep.com/patent/US4397840A ## CPC - C07K 14/505: Erythropoietin [EPO] - A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) - Y10S 530/834: Urine; urinary system - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS ## Abstract The invention provides a novel erythropoietin product prepared from the urine of healthy human but exhibiting no inhibitory effect against erythropoiesis as is the most disadvantageous problem in the product prepared from the urine of healthy human. The method for the preparation of such a product comprises adsorbing the crude erythropoietin product obtained from the urine of healthy human on to a weakly basic anion exchanger from a neutral or weakly acidic aqueous solution containing 0.1 to 0.2 mole per liter of an inorganic neutral salt such as sodium chloride and then eluting the thus adsorbed ingredients with an aqueous eluant solution containing from 0.5 to 0.7 mole per liter of an inorganic neutral salt. ## Claims ### Claim 1 A method for the preparation of an erythropoietin product having no inhibitory effect against erythropoiesis which comprises the steps of (a) adsorbing a crude erythropoietin product obtained from the urine of healthy human on to a weakly basic anion exchanger from a neutral or weakly acidic aqueous solution in the presence of an inorganic neutral salt in a concentration in the range from 0.1 to 0.2 mole per liter, and (b) eluting the thus adsorbed erythropoietin product with an aqueous eluant solution containing an inorganic neutral salt in a concentration in the range from 0.5 to 0.7 mole per liter. ### Claim 2 (depends on 1) The method as claimed in claim 1 wherein the inorganic neutral salt used in step (a) is selected from the group consisting of sodium chloride, potassium chloride, magnesium chloride, calcium chloride and sodium bromide. ### Claim 3 (depends on 1) The method as claimed in claim 1 wherein the weakly basic anion exchanger is a diethylaminoethylated carbohydrate. ### Claim 4 (depends on 3) The method as claimed in claim 3 wherein the diethylaminoethylated carbohydrate is selected from the group consisting of diethylaminoethyl cellulose, diethylaminoethyl dextran and diethylaminoethyl agarose. ### Claim 5 (depends on 1) The method as claimed in claim 1 wherein the neutral or weakly acidic aqueous solution has a pH in the range from 6.0 to 7.5. ### Claim 6 (depends on 1) The method as claimed in claim 1 wherein the aqueous eluant solution had a pH in the range from 6.0 to 7.5. ### Claim 7 An erythropoietin product having no inhibitory effect against erythropoiesis with a molecular weight in the range from 4,000 to about 13,000 and an isoelectric point in the range from 3.1 to 3.6.