Preparation Of Compound Ketconazole Nanoemulsion And Its Pharmacodynamics

In this study, we made nanoemulsion as the carrier of the compound of KCZnanoemulsion. It’s composed of Ketoconazole （KCZ）, Clobetasol Propionate （CP） andEugenol, selected prescription by pseudo-ternary phase diagram, prepared compound KCZnanoemulsion. Investigated its stability, safety and validity, providing a theoretical basis forthe application of the compound KCZ nanoemulsion in the veterinary clinic.1. The preparation of the compound KCZ nanoemuision and its quality evaluationWe used pseudo-ternary phase diagram to investigated nanoemulsion formation factors,screened the prescription of the compound KCZ nanoemulsion, identified the structure typeby staining; examined its shape and the particulate radius distribution by transmission electronmicroscopy and laser particle size analyzer, determined its stability by acceleration tests, lightstability tests and temperature stability tests. Weight percent of each component of thecompound KCZ nanoemulsion were: EL-4024.29%, ethanol4.91%,2-phenylethyl alcohol4.17%, ethyl acetate7.33%, KCZ1%, CP0.05%, eugenol1%, water57.25%, the type of thecompound KCZ nanoemulsion was oil-in-water and it’s liquid was amber, transparent andclear, the drop of nanoemulsion presented spherical in transmission electron microscope,uniform distribution, mean grain size of the drop was11.8nm, and polydispersity index was0.085, with good stability.2. Establishment of the compound KCZ nanoemulsion analysis methodWe used high performance liquid chromatography to established KCZ, CP, and eugenolcontent detection analysis method. The results showed that KCZ, CP, eugenol standard curveat60²20μg·mL^-1,2¹0μg· mL^-1,5¹65μg· mL^-1concentration range had a good linearrelationship, The average recoveries were （99.69±1.14）%, RSD=1.14%,（98.60±1.69）%,RSD=1.71%,（99.82±0.34）%, RSD=0.34%, with-in-day precision and day-to-day precisionRSD were0.57%and1.41%,0.66%and1.38%,0.56%and1.36%. The results showed thatrecovery and precision of the compound KCZ nanoemulsion analysis method were meet therequirements, it can provided a good exclusive method for its preparation and investigation ofstability. 3. Safety evaluation of the compound KCZ nanoemulsionWe used skin acute toxicity test, skin irritation test and skin allergy test to evaluate thesafety of the compound KCZ nanoemulsion. The results showed that there were no acutetoxicity in bandicoots when they were applied with large doses of the compound KCZnanoemulsion, there were no obvious irritant when intact skin and damaged skin of the testedanimals were applied with the clinical doses of the compound KCZ nanoemulsion, the testedanimals without an allergic reaction. The compound KCZ nanoemulsion transdermal drugdelivery safety, little irritation.4. The effect of transdermal in vitro of the compound KCZ nanoemulsionWe used bandicoot skin and the transdermal diffusion instrument to detect the compoundKCZ nanoemulsion transdermal effect. Compared the effect of transdermal in vitro of thecompound KCZ nanoemulsion, compound KCZ nanoemulsion with2.5%azone, compoundKCZ nanoemulsion with5%azone, compound KCZ ointment,1%ketone itraconazolesuspension,0.05%CP suspension,1%eugenol suspension. The results showed that therewere no significant difference between the compound KCZ nanoemulsion and plus2.5%azone compound KCZ transdermal rate, both of them were higher than plus5%azonecompound KCZ nanoemulsion, Compound KCZ nanoemulsion transdermal effectsignificantly better than the suspension of the main component, KCZ and CP in the compoundKCZ nanoemulsion transdermal effect were better than which in compound KCZ ointment.The compound KCZ nanoemulsion has a good effect on transdermal drug delivery.5. The drug effect of the compound KCZ nanoemulsionWe detected the minimum inhibitory concentration （MIC）, the minimum fungicidalconcentration （MFC） and the sterilization time of the compound KCZ nanoemulsion toSaccharomyces cerevisiae and Candida albicans test to investigate in vitro antibacterial effectof the compound KCZ nanoemulsion. The results showed that MIC of the compound KCZnanoemulsion, KCZ suspension, clotrimazole suspension against Candida albicans andSaccharomyces cerevisiae were0.5,0.5μg·mL^-1,4,4μg·mL^-1,8,32μg·mL^-1, blanknanoemulsion and sterile saline had no inhibitory effect, Candida albicans andSaccharomyces cerevisiae under the influence of8μg·mL^-1KCZ suspension for3h, showedno growth, under the influence of1μg·mL^-1compound KCZ nanoemulsion for1.5h, showedno growth, under the influence of64μg·mL^-1clotrimazole suspension for4h, two kinds offungi showed no growth. The compound KCZ nanoemulsion has a good in vitro antibacterialeffect on these two fungi.We separated rabbit itch mites and used different drugs to kill them to investigate the compound KCZ nanoemulsion in vitro acaricidal effect, recorded the time of death. Theresults showed that the acaricidal time of the compound KCZ nanoemulsion was6⁷min, andno recovery, the acaricidal time of XUAN MAN JING and CHU MAN LING sprays, were15¹7min,20²1min, and there were some mites recovered, blank nanoemulsion anddistilled water have no acaricidal effect. the compound KCZ nanoemulsion have a good invitro acaricidal effect.We cured dogs infected with fungal skin disease with the compound KCZ nanoemulsion,compound KCZ ointment and clotrimazole ointmentto to investigate in vivo antibacterialeffect of the compound KCZ nanoemulsion. The results showed that cure rates were96.7%、76.7%and73.3%, effective rates were100%、83.3%and80.0%. We cured dogs infected withmite disease with the compound KCZ nanoemulsion, XUAN MAN JING and CHU MANLING sprays to investigate the compound KCZ nanoemulsion in vivo acaricidal effect. Theresults showed that cure rates were96.7%、70.0%and76.7%, effective rates were100%、80.0%and80.0%. The compound KCZ nanoemulsion has a good clinic antibacterial andacaricidal effect.The prepared compound KCZ nanoemulsion had a high safety, good efficacy, and goodstability, which could provide a new type of drug for veterinary clinical medicine.