Stduy Recombinant Human Granulocyte Macrophage Colony-stimulating Factor Spray

Recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) is a modern biotechnology product. It is a non-glycosylatedproteincomposed of127amino acids, and it is soluble in water. rhGM-CSFrestores hematopoiesis of bone marrow granulocytes, promotes proliferationand differentiation of hematopoietic cells,, increasesperipheral white blood cells, and stimulates maturationofneutrophils and mononuclear macrophages. It’s now widely used in the treatment of severe leucopenia in leukemia and other cancer patients after high dose chemoradiotherapy. In recent years, it is widely used externally in clinic. The main administration method is to dissolve the injection powder in saline and directlyapplyon the affected parts by means such as gargling andwipingwith cotton. However, the absorptionand utilization of the drug is poor this way, and the remaining drug liquid cannot be saved once the drug vial is opened. In order to protect rhGM-CSF activities in liquid over a long period of time and ensure the drug stability, safety and effect, we developed rhGM-CSF spray and studied its pharmacodynamics. The results are as the following:1. We completed the rhGM-CSFspray formulation. We screened and optimized protein protectants, transdermal absorbents, antibacterial agents and phosphatebuffer systems. We determined the best formulation with the minimal impact on protein activity and the best drug effect after opening the bottle.2. We determined the quality standards of the rhGM-CSFspray. The stability test results show that the protein activity dropped10.12%onaverage after investigation at37±2℃condition for30days, dropped8.04%on averageafter investigation at25±2℃condition for12months, dropped4.03%onaverage after investigation at2-8℃condition for12months, and dropped1.64%onaverage after investigation at4500LX±500LX illuminance for10days at room temperature. Based onthe2010third edition of Chinese Pharmacopoeia, we established the relatedquality standards:appearance as light-yellow clear solution, total press times not less than100, total spray volume0.08-0.12ml, filling volume not less than9.5ml, pH value6.0-7.0, identification as positive, sterilization test qualified and biological activities detection range1.6×104IU/ml-3.0×104IU/ml.3. We finished therhGM-CSFspray toxicology studies.The results show that it is safe and reliable to use rhGM-CSFspray externally. We used the800μg/mlrhGM-CSF stock solution to conduct acute toxicity test on rabbits. The rabbits remained normal after drug administration, and it demonstrated non-toxic side effects of the drug. We used20μg/ml spray to do skin allergy test in guinea pigs and irritation test on rabbits.The tested animals didn’t showallergy or irritation, which proved the external useof rhGM-CSF spray to be safe. 4. We completed the pharmacodynamicstudies of the rhGM-CSFsprayonburns,scalds, skin ulcers, and oral ulcers. We used different concentrationsof rhGM-CSF spray to dose the shallow Ⅱ degree burn experimental model of rabbits, the shallow Ⅱ degree scald experimental model of rabbits, the skin ulcer experimental model ofguinea pigs, and the oral ulcer experimental model of mice. It shows that there is obvious therapeutic effect in the rhGM-CSF treatment group on burns, scalds, skin ulcers, and oral ulcers.Through the above rhGM-CSF spray pre-clinical studies of formulation screening, quality standards and toxicology, the research provides significant early-stage work foundation to further clinical studies of the rhGM-CSF spray...