Study On The Process Of Recombinant Human Follicle Stimulating Hormone Biosimilar Drugs

Recombinant human follicle stimulating hormone(rhFSH)is a drug which can stimulate ovarian follicular growth in women.Its most effective function is to stimulate the follicles mature.FSH can efficiently recover ova and oocytes,to gain the viable fertilized ova.The results of clinical studies shows that the pregnancy rate of the pregnant women and the baby’s birth and the survival rate is very high after using rhFSH.At its earliest stages,most injections were made with follicle stimulating hormone,which is mainly used in patients who do not ovulate in order to stimulate the development of individual follicles and thus stimulate ovulation.At present,most manufacture processes of the domestic products used the backward production technology,the cost of production is very high,and the adverse reactions and side effects of the products are very big,so compared with other foreign varieties,the competitiveness of these products is declining in gradually year by year,and then low profits.So the state encourages new products to replace them.Currently,there are many common drugs to promote ovulation,including gonadotropin,aromatase inhibitor and dopamine receptor agonist.As one of the dominant varieties of gonadotropin,rhFSH occupy a very heavy market share,and is applied more widely.,a product of recombinant human follicle stimulating hormone,manufactured by Serono enter the market of China in 2002.Because the domestic market demand is very high,Gonal-F has a good market prospects.Thus the market potential of developing human recombinant follicle-stimulating hormone biosimilar drugs is extremely high.In our country,compared with other developed countries,the biosimilar drugs development still exists a certain gap,the most products used in domestic are from abroad.This is especially true for rhFSH manufacture.In recent years,the market demand for human FSH has increased greatly,so the developments of rhFSH products are particularly important for creating our own brand and changing the situation of foreign control.Based on the multiple perspective of rhFSH,such as the basic overview,the structure,the physical and chemical properties,and the problems existing,our group conduct a deeper study on the manufacture processes,the properties and the functions of rhFSH.The main contents of this study include selecting suitable cells for rhFSH expression,developing an efficient and stable protein purification process to ensure the purity and the recovery of rhFSH,and establishing a suitable quality specification for rhFSH.The main test items include the content of protein,the biological activity,the purity,the molecular weight,the residues of exogenous DNA residues,the host cell protein level,the bacterial endotoxin level,the sialic acid content,the isoelectric point and the charge heterogeneity,the ultraviolet spectrum and peptide mapping,etc.Purification process research is the key and difficult point for this subject,the complexity of the purification process is determined by the characteristics of the protein,and the main steps of the purification process established in this study are as follow: the cell culture supernatant → affinity chromatography and high pH virus inactivation → hydrophobic chromatography → CHT chromatography → ionexchange chromatography → ultrafiltration concentration → virus filtering and aseptic filtration → the bulk solution.The final yield of the process is more than 50%.In addition,the problem on which rhFSH is unstable under acid condition is solved,and a high purity is gained after the purification process because this process can remove the impurities including medium residue,viruses,oligomers,low molecule weight degradation substances,host cell protein,nucleic acid,endotoxin,etc.The purity of the bulk solution is higher than 99.5%,and the product quality is better than the original product.