Listed Species "Kangfuxin" Pharmaceutically Re-evaluation

Peptic ulcer (peptic ulcer, PU) is a frequent disease and common disease. Kangfuxin liquid is the single preparation, made by Periplaneta americaana (L.) extract, oral administration for the treatment of peptic ulcer in clinical. In this thesis, based on ancient herbal medicine, departure from the clinical efficacy, around the Kangfuxin liquid’s substance inside and outside anti-peptic ulcer pharmacodynamic differences of the Production, combined material basis for efficacy in the process and related mechanisms, quality assessment, we carry out the listed species Kangfuxin liquid pharmaceutically re-evaluation system, for the clinical use of safety, efficacy, and its secondary development to provide a theoretical basis and technical support.1.The process reviewThe actual production process as the core, the efficacy (The rat pylorus ligated gastric ulcer model and Water immersion stress gastric ulcer model) combined with chemical analysis to scientific rationality re-evaluation of the existing process of Kangfuxin liquid. The results showed that:the existing process which removed special fishy smell of grease didn’t have the role of prevention and treatment of peptic ulcer, actually worsened the extent of the ulcer; the crude polysaccharide and protein had not been extracted in the process also did not show anti-ulcer activity; anti-peptic ulcer active ingredient was still concentrated in Kangfuxin liquid. Thus initially clarify the Kangfuxin liquid raw materials Periplaneta americaana (L.) active substance of anti-peptic ulcer had no relate to the fats, polysaccharides, and high molecular weight protein. So the existing Kangfuxin liquid production process has a certain scientific rationality.2.Element analysis and pharmacodynamic evaluation of each eluate of Kangfuxin liquid intermediateAccroding to Hydrophobic strength of different Kangfuxin liquid intermediate composition, the use of macroporous resin, gradient elution, Obtained water eluate,25%ethanol material,50%ethanol elution and75%ethanol eluate four parts, Combined with chemical and efficacy (Gastric ulcer model of water stress) experiment carried out by activity tracking and component analysis of each eluate of Kangfuxin liquid intermediates. Results:Kangfuxin liquid intermediates were separated and the water eluate did not play against peptic ulcer, actually worsened the extent of the ulcer, suggesting that the amino acids, carbohydrates, composite nucleotide bases constituents and epidermal growth factor in the water of eluate had little to do with anti-peptic ulcer effect;25%ethanol elution had trend to prevention and treatment of peptic ulcer; better prevention and treatment of ulcers in50%ethanol elution; efficacy of75%ethanol ulcer was the most significant, and better effective than before purification, showed that the active substance of the Kangfuxin liquid to anti-ulcer was mainly concentrated in75%ethanol elution, and the purity of the peptide component up to90%, suggesting that Kangfuxin solution’s anti-peptic ulcer effect is related to peptide component.3.Efficacy mechanismBy enzyme linked immunoassay (Elisa method) detected the content of5-hydroxytryptamine (5-HT), dopamine (DA) and norepinephrine (NE) in serum; Immunohistochemical techniques detection the number of heat shock proteins (HSPs), H+/K+-ATPase enzyme, prostaglandin (PGE2), epidermal growth factor (EGF) in gastric tissue, we explored the antiulcer mechanism of Kangfuxin solution and its elution. Results:In the stress ulcer, HSPs and EGF did not reflect the protection role of gastric tissue. The water eluate didn’t have anti-ulcer effect and even increased the formation of ulcers and inflammation, caused by failing to inhibit H+/K+-ATPase activity and stimulate PGE2inflammatory mediators release. Kangfuxin liquid antiulcer’s action is based of the inhibition of5-HT, NE release and H+/K+-ATPase enzyme activity, and inhibition of PGE2synthesis;25%ethanol elution had a trend to have antiulcer effect, it is possible to suppress the5-HT, NE, and et al;50%ethanol elution was more stable in anti-ulcer effect, and it had a strong relate to the inhibition of5-HT, NE secretion and of PGE2synthesis;75%ethanol elution antiulcer action was most significant in anti-inflammatory effects, not only can inhibit the secretion of5-HT and NE, but also are able to significantly inhibit the H+/K+-ATPase enzyme activity and inhibition of gastric acid secretion, and inhibition of PGE2synthesis to reduce the occurrence of inflammation and promote DA elevated levels et al. These factors played a significant role in anti-ulcer. At the same time, each eluted material’s anti-ulcer efficacy had a good repeatability.4.Composition and structure identification of the Kangfuxin liquid intermediate75%ethanol elutionComponent of75%ethanol elution were analysised by GC-MS, the results suggested that remove of the grease in Kangfuxin liquid production process and the preparation of75%ethanol elution is the key link, and removing of grease incomplete will remain on the relative percentage higher levels of palmitic acid with cytotoxicity; toxic substances of macroporous resin do not comprise in the75%ethanol elution, so it can be used safely. Using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS), results of the the determination of the molecular weight of the peptide component of the75%ethanol elution showed that the molecular weight distribution between500-3300substances. m/z520.34maximum response was the mainly substance. In addition, there existed some polypeptide component, such as m/z1348.7432, m/z1943.0382et al.5.The quality review mode of Kangfuxin liquid and its raw materialsHPLC methods was used to establish fingerprint of Kangfuxin liquid and its bulk drugs Periplaneta americana herbs, and the fingerprint pre-treatment is simple, informative, as well as omprehensive evaluate of the herbs and preparations. Results: there were differences among the Fingerprint of Periplaneta americaana (L.) from Columbine medicine, Lotus Pond Chinese herbal medicine market and production bases in Yunnan; the Periplaneta americaana (L.) herbs was different from Periplaneta fuliginosa at characteristic peaks, which can be used as a basis for identification;10batches of Kangfuxin solution of A pharmaceutical production were more uniform and stable except several batches.On the basis of establishment medicinal herbs and preparations fingerprint combined with efficacy findings, used the fingerprints of severe ulcers water elution from a comparative analysis of10batches of Periplaneta americaana (L.) herbs fingerprints and Kangfuxin liquid fingerprints Total mode, got total peaks of water eluate, herbs and preparations. After striking out the total peaks, the herbs and preparations’HPLC fingerprint similarity was evaluated and the total peak cluster was analysised, and used the Columbine medicine Periplaneta americaana (L.) herbs, having anti-ulcer effect, lot for M1204122, Kangfuxin liquid as a reference. The results showed that the Periplaneta americaana (L.) of Lotus Pond Chinese herbal medicine market grouped separately as a class, the Columbine medicine and production bases in Yunnan Periplaneta americaana (L.) herbs classified as a category, suggesting that besides the Periplaneta americaana (L.) comes from the Lotus Pond Chinese herbal medicine market, other herbs in the anti-ulcer had a good agreement;10batches of A Pharmaceutical production Kangfuxin liquid although divided into two categories, but the similarity and correlation coefficient of the batch M1204122Kangfixin liquid was very high with other batches, indicating that the A Pharmaceutical production Kangfuxin liquid anti-ulcer effect is more stable.We re-evaluate the Kangfuxin liquid from the perspective of the physical properties by measuring Kangfuxin liquid’s conductivity, surface tension, osmotic pressure, viscosity and other parameters,. The results showed that the quality of Kangfuxin liquid was also stable.In summary, the material basis and mechanism of the Kangfuxin liquid anti-peptic ulcer was more specific. Based on the evaluation model of exclude the invalid parts of the component peak of the fingerprint and the physical and the quality re-evaluation mode of chemical parameters characterize, We provided a new approach for quality evaluation. The existing production processes of Kangfuxin liquid had certain reasonable and room for growth, the role of prevention and treatment of peptic ulcer can also be significantly improved. For this reason, we suggested that Kangfuxin liquid production or secondary development process should remove grease as much as possible and monitor the content of palmitic acid, and enrich peptide ingredients to fully guarantee the effectiveness and quality of clinical controllability.