A group of gene recombinant human corticotropin precursors for improving serum glucocorticoid levels and preparation method

Abstract

The present invention relates to four kinds of gene recombinant human corticotropin (ACTH) precursor proteins with the effect of increasing the level of glucocorticoid in serum: ProACTH113, ProACTH72, ProACTH80, ProACTH141, and a preparation method of the four precursor proteins. ProACTH113 contains the polypeptide chain of amino acids 105-217 of Pro-opiomelanocortin (POMC), ProACTH72 contains the polypeptide chain of amino acids 105-176 of POMC, ProACTH80 contains the polypeptide chain of amino acids 138-217 of POMC, ProACTH141 contains the polypeptide chain of amino acids 77-217 of POMC; the four ACTH precursor proteins all contain the amino acid sequence of amino acids 1 to 39 of ACTH polypeptide. To facilitate purification, the N-termini of four precursor proteins, such as ProACTH113, ProACTH72, ProACTH80, and ProACTH141, were linked to enterokinase substrate sequence (EKst), and enterokinase with a histone tag was used to excise the N-terminal of these four precursor proteins in vitro. The excess peptides attached to the ends generate free ProACTH113, ProACTH72, ProACTH80, and ProACTH141, and then use a nickel column to remove the enterokinase linked with histone tags.

Description

technical field [0001] The invention belongs to the field of genetic engineering and protein engineering pharmacy, and specifically relates to a group of corticotropin precursor proteins (ProACTH) and their DNA and protein sequences, as well as a preparation process of the group of precursor proteins. Background technique [0002] Corticotropin (Corticotropin), also known as adrenocorticotropic hormone (ACTH), is a 39 amino acid polypeptide hormone secreted by basophils in the anterior pituitary gland. Biosynthesis and metabolism of fat and protein, regulate cardiovascular and cerebrovascular functions, improve resistance, protect and regenerate nerve damage, anti-inflammatory, immunosuppressive, anti-toxin, anti-shock, etc. ACTH is the only effective drug for the clinical treatment of convulsions in infants under two years of age. The incidence of the disease is 0.5% to 1%, 6 to 33% of infants with convulsions die before the age of 3, and 70 to 90% of children with convulsi...

AI Technical Summary

Problems solved by technology

At present, ACTH for domestic clinical application is mainly extracted from animal tissues, the production cost is relatively high, and there is a potential risk of spreading animal viruses and mycoplasma

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