Pharmaceutical Reasearch On KGJS Ointment And The Evolution Using Zebrafish

This article inclued two part,part one:pharmaceutical reasearch on KGJS ointment and part two:Elucidated the anti-osteoporosis material basis of Epimedium based on M-ACT/TOX technology by zebrafish.Part one:pharmaceutical reasearch on KGJS ointmentKGJS ointment,a prescription of orthopedics in Jiangsu Province Academy of Traditional Chinese Medicine,is used to treat knee and lumbar osteoarthritis and shoulder periarthritis,which is composed of 10 medicine such as Zanthoxylum bungeanum Maxim.,Artemisiae argyi folium,Atractylodis rhizoma,Angelicae pubescentis radix,Cinnamomi ramulus,Carthami flos and so on.However,it is inconvenience to prepare and store,which limits the usage scope.In order to make the recipe be used for more patients,this study provided an experimental basis for making the prescription to be a modern TCM preparation based on TCM theory.According to the relevant requirements of drug registration management measures of CFDA,we proceed the pharmaceutical research of KGJS ointment,including literature research,preparation technology,quality standard,stability studies,preliminary efficacy studies and the skin irritation of KGJS ointment in rabbits.Besides,as a common medicine of orthopedics,the activity and toxicity of Epimedium are unclear.Hence,this article illustrates the evolution of Epimedium Act/Tox by M-Act/Tox technology using zebrafish.1.Literature researchThe part of introdution expounds Arthritis research progress and summarizes mainly ingredients in the prescription of physical and chemical properties and pharmacological effects of KGJS ointment Treatment.2.The research of preparation technologyBasis of the research of molding powder and preparations,examining index of powder rate and grinding characteristics(grade of grinding difficulty level and adhering on the wall or not),we optimized the best grinding process is the grinding process of whole prescription,according to the classification of separate,sorted and whole mixture crush.After the selection matrix as glycerin and vaseline,based on the index of humidity,viscosity and mixed for examining,prescription test matrix content and ratio,mixing temperature,mixing method of powder and matrix,using the single factor and L9(34)orthogonal expriments.Campared with different preservatives of potassium sorbate,plus sodium benzoate with ethylparaben,prescription are selected the best one as plus sodium benzoate with ethylparaben.After three small-scales and pilot-scale process,final forming process is as follow:ten medicine of KGJS ointment blend and crush into powder,then add 600g glycerin,200g vaseline and 3.3g plus sodium benzoate with ethylparaben(dissolved by water),thoroughly mixed,weight of final ointment about 1100g.3.Quality specification studyIn order to make the product quality is controllable,this paper examined the name,character,identification and standard draft.According to the result of content determination of the three batch samples,the content determination of osthole is not less than 0.60mg per 10g.4.Stability studyIn order to investigate the stability of the drug,to determine the period of validity,according to the traditional Chinese medicine,natural medicine stability study technical guiding principles,this subject tested the factor affecting experiment for the sample of three trial-production.We carried out the accelerated stability test for 6 months and long-term stability test for 6 months.Proceeded the study of the characters,identification and content determination of the products.The result showed that the quality is stable.5.Skin irritation and the evalution of toxicity by zebrafishGiven KGJS ointment 0-72h,rabbit back does not appear any adverse reactions such as redness,congestion,oozy,degeneration or necrosis.Part two:Elucidated the anti-osteoporosis material basis of Epimedium based on M-ACT/TOX technology by zebrafish1.To illustrate the Epimedium activity and toxicity constituents,we exam different kinds of epimedium extract,constituents(YTF,SWJ,FS and DT),ingredient(eight monomers of YTF,one monomer of SWJ and three monomer of FS)and YTF compatible with SWJ,FS or DT based on M-Act/Tox technology,investigating the transformation of metabolism,activity and toxicity.2.Discoveries of evolution of Epimedium anti-osteoporosis activity are as follows:anti-osteoporosis activity can be inspected by different kinds of Epimedium;YTF is the major active component in Epimedium,and the major mechanism is Downregulation of RANKL gene expression;FS has anti-osteoporosis activity;SWJ and DT have no anti-osteoporosis activity.3.Discoveries of evolution of Epimedium toxicity are as follows:with the change of different kinds,Epimedium show different toxicity such as swelling,darkening and distorting of zebrafish yolk sac.Epimedium 50%ethanol extraction expose to model zebrafish more toxic than normal zebrafish.Toxicity of vary components are difference:SWJ and YTF have toxcity tozebrafish and next is FS,but DT is safest.Epimedium monomer shows different toxicity:SB,SC>EC,SA>YG,BI,EA,EB.There are two aspects of toxicty of zebrafish exposed to YTF monomer groups:increasing and decreasing toxicity of metabolite.LYS,XLYS,YLYS and MLHJ show low toxicity.4.Discoveries of zebrafish metabolism of YTF components and monomers are as follows:The metabolic pathways of YTF in zebrafish are similar.It is easy for them to cleave 7-0 glucosides,3-0 glucosides and 3-0 xylopyranose,but it is hard to disconnect rhamnose.So YTF monomers can be translated into SC which contains rhamnose or BI.EA,EB,YG,SB can be primarily translated into BI.EC can be primarily translated into SC.Metabolic rate of model zebrafish faster than normal zebrafish.5.The evolution of influence of different components combine with YTF using zebrafish.DT has the ability to accelerate metabolic biotransformation of YTF obviously and increase toxicity of YTF.SWJ has less effect to accelerate metabolic biotransformation of YTF,but increase toxicity of it.There is no significant anti-osteoporosis activity of DT,FS and SWJ.