Preparation And Quality Study Of Snake Venom Protein Suppositories

1 ObjectiveThe purpose of the experiment is to prepare snake venom protein suppository.To establish correlation analysis and determination methods,to optimize the formulation and preparation process of snake venom protein suppositories,to conduct quality evaluation.At the same time,the analgesic effect and pharmacological activity of snake venom protein suppository were investigated.2 Methods(1)Establishing the an analysis method of venom protein in vitro,dissolution method of snake venom protein suppositorty in vitro,making a systematic test of it.(2)Optimizing and determing the prescription and preparation process of snake venom protein suppository by single factor and orthogonal experiment and preparing snake venom protein suppository.Conducting quality assessment.(3)Using the hot plate method and the acetic acid writhing method to perform an analgesic experiment of snake venom protein suppositorty.The experiment is divided into five groups,including the blank group,the the high-dose group(120 μg/kg),middle-dose group(60 μg/kg)and low-dose group(30 μg/kg).3 Results(1)High performance chromatography(HPLC)method was established to determine the analysis method of sanke venom protein in vitro.The linearity of snake venom protein was in the range of 3.5 to 350 μg/mL(r=0.997);the precision,accuracy,recovery and stability were all fit to methodological verification requirements.High performance chromatography(HPLC)method was established to determine the dissolution rate of snake venom protein in vitro.The linearity of snake venom protein was in the range of 3.5 to 350 μg/mL(r=0.999);the precision,accuracy,recovery and stability were all fit to methodological verification requirements.(2)The optimal prescription of snake venom protein suppository were as follows:PEG 400 2g,PEG 4000 5g,temperature 50℃,2.5%tween.The suppository prepared under the optimal prescription is smooth and uniform in appearance with moderate hardness.The melting time of thawed suppository melted within 60 min,the content was as high as 85%and the dissolution rate reached 80%,the content uniformity conforms to regulations.(3)The result of the acetic acid writhing test show that compared with the blank group,there was a significant difference in the number of writhing in each group of snake venom protein suppositories;the number of writhing in the pethidine group was 7.13 ±3.76(P<0.05),The analgesic effect of medium and high doses of snake venom protein suppositories was quite similar to pethidine group,and the number of writhings in medium and high doses of snake venom suppositories was 8.88±1.46 and 4.75±3.10,respectively(P<0.05).The result of the acetic acid writhing test shows that the high and middle doses of snake venom protein suppositories all significantly prolonged the pain threshold in the latency period of pain response to thermal stimulation in mice,and the pain threshold was significantly prolonged.The high dose of snake venom protein has the best effect.Compared with the blank group,the difference in pain threshold was statistically significant(P<0.05).Compared with pethidine group,the analgesic effect was similar in the high-dose group(P>0.05);the middle and low-dose groups were all inferior to pethidine group,which was statistically significant(P<0.05).Conclusion:The snake venom protein suppository prepared in this study meets the requirements and can be rapidly released in vitro.The above studies show snake venom protein suppository has good analgesic effect.