METHOD OF PRODUCTION OF GRAFT CO-POLYMER EXCIPIENT WITH A SUPERIOR PEPTIDE AND PROTEIN BINDING PROPERTY

摘要

Provided herein is a process of preparing a semi-random graft co-polymer, the product of which is difficult to fully characterize chemically. The product of the present disclosure has unique and useful properties of 1) binding to a peptide and 2) upon co-administration of the product and the peptide into animals the product prolongs the blood circulation time and elevates the level of the peptide, compared to the peptide alone without the product of the disclosure.

主权项

1. A method of preparing a semi-random graft co-polymer comprising the steps of:
(a) dissolving a linear polyamine backbone containing W moles of free primary amino groups in an aqueous buffer having a buffering range from pH 7 to pH 8 to provide solution A having a volume Y; and conducting either B1 or B2, wherein:
B1 comprises (b) activating a protective chain containing 0.5-1.2×W, 0.5-1.1×W, or 0.5-1.0×W moles of carboxyl groups with 1.7-7.0×W, 1.7-3.7×W, or 1.7-3.4×W moles of N-hydroxysuccinimidesulfate and 1.5-3.6×W, 1.5-3.3×W, or 1.5-3.0×W moles, respectively, of 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride in an aqueous buffer, at pH 4-5.5 in a final volume of Z, and at a W/(Y+Z) ratio of 30-55 mM, and allowing the activation to proceed for a period of time between 0-30 min to provide solution B; (c) adding solution B to solution A with continuous mixing to provide solution C; and (d) adjusting the pH of solution C to above 6.5; and
B2 comprises (b-d) adding a protective chain containing 0.5-1.2×W, 0.5-1.1×W, or 0.5-1.0×W moles of carboxyl groups, 1.7-7.0×W, 1.7-3.7×W, or 1.7-3.4×W moles of N-hydroxysuccinimidesulfate, and 1.5-3.6×W, 1.5-3.3×W, or 1.5-3.0×W moles of 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride to solution A to provide solution C; and (e) adding 0.5-1.5×W, 0.5-1.2×W, 0.5-1.1×W, or 0.5-1.0×W moles of additional 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride to solution C after 2 to 3 hours and waiting until the primary amino groups are 55-40% of W moles.