Preparation And Pharmacodynamics Study Of Compound Vaginitis Nanoemulsion In-situ Gel

Vaginitis was a common and frequently-occurring gynecological disease,which was a collective name of vaginal mucosa diseases caused by a variety of pathogenic microorganisms.Among such diseases,trichomonas vaginitis took up 13%~25%,bacterial vaginitis 10%~30%,fungal vaginitis 40%~50%.Purpose: This research aimed to develop a new preparation of treatment value to all kinds of vaginitis with relatively high therapeutic effects.To be specific,first,develop the drug-containing nanoemulsion,optimizing nanoemulsion so that it achieved transparency and stability,with active pharmaceutical ingredients included as Terbinafine and Levornidazole;then swell the gels with the prepared drug-containing nanoemulsion as the solvent.The gels used were mainly P188 and P407.By adjusting the proportion of the two gels,develop a preparation that remains liquid at room temperature and semi-solid at body temperature.Method: Draw with the pseudo-ternary phase diagram method to guide the preparation of nanoemulsion;add Terbinafine and Levornidazole into the nanoemulsion to prepare the drug-containing nanoemulsion.Terbinafine and Levornidazole shall be determined through the following tests: antimicrobial susceptibility testing in vitro with antifungal drugs fluconazole,miconazole,ketoconazole and terbinafine on Candida albicans,Candida parapsilosis and Candida glabrata respectively;antimicrobial susceptibility testing in vitro with drugs against anaerobic bacteria and trichomonas metronidazole,tinidazole,secnidazole and Levornidazole on bacteroides fragilis,Escherichia coli and Gardnerella vaginalis respectively.Then,use central composite design-response surface methodology to optimize the nanoemulsion preparation process,use δ potential instrument to measure the particle size,particle distribution and the δ potential of drug-containing nanoemulsion,and use transmission electron microscope to detect the morphologies of the particles;test and research repeatedly with P188 and P407 as basic gel compositions to determine the proportion of gel states that satisfy the requirements;swell up the gels with the optimized drug-containing nanoemulsion as the solvent till the drug-containing nanoemulsion meet the requirements under the objective.Conduct stability study of the prepared drug-containing nanoemulsion,including study on high-temperature stability,high-humidity stability and light stability;for the study of safety,conduct study of irritation on the vagina of rats;for the test on effectiveness,perform antimicrobial susceptibility testing in vitro with Terbinafine and Levornidazole-containing nanoemulsion gel on Candida albicans,Candida parapsilosis,Candida glabrata,bacteroides fragilis,Escherichia coli and Gardnerella vaginalis respectively to verify its effectiveness.Results: The author selected Terbinafine,which presented the best effects from the antimicrobial susceptibility tests in vitro with 4 antifungal drugs on 3 fungal pathogens,as the active ingredient against fungi in this research,and;Levornidazole,which presented the best effects from the antimicrobial susceptibility tests in vitro with 4 drugs against anaerobic bacteria and trichomonas on 3 anaerobic bacteria,as the ingredient against anaerobic bacteria and trichomonas in this research.The Terbinafine and Levornidazole-containing nanoemulsion prepared in this research was in liquid state with high fluidity,clear,transparent and in the color of light yellow;the average particle size measured by the δ potential instrument was 11.5 nanometers,with δ potential at-13 m V.The nanoemulsion formulation optimized with the central composite design-response surface methodology was as follows: Terbinafine 3.4g,absolute ethyl alcohol 38.7g,Ornidazole 1.7g,IPM10.4g,tween-80 100.2g and purified water as the remaining part,totaling an amount of 1000 g.Results of transmission electron microscope testing indicated that,the basic particle morphology was a regular circle.In the prepared gel,P188 and P407 were proportioned as: P188 18 g,P407 100g~110g.The prepared drug-containing nanoemulsion gel was in liquid state with good fluidity at room temperature,and in semi-solid gel state at body temperature.Vaginal irritation testing indicated no irritation.Results of effectiveness test demonstrated that,the inhibition zone of the drug-containing nanoemulsion gel against Candida albicans,Candida parapsilosis and Candida glabrata reached 37.±3.4mm,27.2±2.5mm and 31.6±4.1 mm respectively.Results of antimicrobial susceptibility tests in vitro on bacteroides fragilis,Escherichia coli and Gardnerella vaginalis revealed that,no bacterial colony was formed on all of the drug-containing media.Conclusion: The drug-containing nanoemulsion gel prepared in this research for prevention and treatment of vaginitis was safe,stable,effective and non-irritating.