Preliminary Preformulation Pharmacy Research Of Beauvercin And Development Of Ketoconazole And Beauvericin Combination Drug Tablets

Fungal infections disease is a common clinical disease, which can be divided into superficial fungal infections disease and deep fungal infections disease according to the depth of the fungal invasion. Between two of them, deep fungal infections disease is more harmful to human health, and the mortality is very high. Ketoconazole is a synthetic broad-spectrum antifungal agent with the risk of hepatoxicity. However, it was found that the combination use of ketoconazole and beauvericin in their low dose can not only maintain the antifungal activity of ketoconazole but also significantly reduce the liver toxicity of ketoconazole.Based on these researches, the ketoconazole and beauvericin combination drug products were expected to be developed as a state first class new medicine. This article is focus on the preliminary preclinical pharmacy research of ketoconazole and beauvericin combination drug products. The study is composed of two parts. (1) Preliminary preformulation pharmacy research of beauvericin, include:development and validation of analysis methodology, structure confirmation, physicochemical properties, preliminary stability studies and preliminary quality standard research of beauvericin. (2) Development of the ketoconazole and beauvericin combination tablets, include:compatibility study of drug and excipients, formulation screening, preparation process, development and validation of analysis methodology, preliminary stability studies and preliminary quality standard research of ketoconazole and beauvericin combination tablets.The structure of API (Active Pharmaceutical Ingredient) was confirmed by IR, MS,1H-NMR,13C-NMR. The results indicated that the structure of the API compound is corresponding with the structure of beauvericin.The appearance, melting point, solubility and the property of absorb moisture of beauvericin have been studied. The results indicated that beauvericin is white amorphous powder. It can absorb moisture, slightly. The melt point is at 147～149 ℃. It is almost insoluble in water. The oil-water partition coefficient was measured by using octanol and water, and the results showed that almost all of beauvericin was distributed into the layer of octanol, which indicated that beauvericin is highly lipophilic.We have developed a gradient HPLC analytical method with Synergi Fusion-RP column used for the stability studies of beauvericin. The method validation results indicated that the new HPLC method could be successfully applied in the stability studies of beauvericin. The forced degradation testing results indicated that beauvericin is unstable under alkaline condition.The purity, loss on drying, residue on ignition, heavy metals and residual solvents of beauvericin API have been studied in this article. Ethanol, acetone, acetonitrile, isopropyl ether and ethyl acetate were used as solvents in the preparation process of beauvericin API. We developed a head-space gas chromatography method for the determination of residual organic solvents in beauvericin API. The method validation results indicated that the head-space gas chromatography method could be successfully applied in the residual organic solvents studies of beauvericin.The stability of the beauvericin was evaluated by impress factor testing and accelerated testing. The impress factor testing indicated that high-temperature and high-light exposure conditions had no significant impact on the stability of beauvericin. The high-humidity testing indicated that beauvericin absorb moist slightly, which hint that beauvericin should be kept under dry conditions. The accelerated testing results indicated that the appearance, content and related impurity of beauvericin had no obvious change within three months.On the basis of quality research of the beauvericin API, the quality standard was developed, include properties of appearance, identification, content determination, related substances, residual solvents, loss on drying, residue on ignition and heavy metals.The ketoconazole and beauvericin combination drug products were developed in our research.We have developed a gradient HPLC analytical method with Synergi Fusion-RP column used for the stability studies of ketoconazole and beauvericin combination drug products. The method validation results indicated that the new HPLC method could be successfully applied in the stability studies of ketoconazole and beauvericin combination drug products. The forced degradation testing results indicated that the mixture of ketoconazole and beauvericin were unstable under acidic, alkaline, oxidizing and light condition.A gradient HPLC analytical method with Luna C18 column used for determination of ketoconazole and beauvericin content in combination drug tablets have been developed. The method validation results indicated that the new HPLC method could be successfully applied in the determination of ketoconazole and beauvericin content in combination drug tablets.Several dissolution mediums were screened and 0.5% SDS solution was selected as dissolution medium, finally. A gradient HPLC analytical method with Luna C18 column used for the dissolution rate determination in combination drug tablets has been developed. The method validation results indicated that the new HPLC method could be successfully applied in the dissolution rate determination in combination drug tablets.After online searching from "U.S. FDA Drugs Database", we learned that the excipients of ketoconazole tablet which already on the market from Johnson & Johnson were microcrystalline cellulose (MC), lactose, corn starch, polyvinyl pyrrolidone (PVP K-30) and magnesium stearate. On the basis of the literature, the formulation and preparation process were optimized.The stability of ketoconazole and beauvericin combination drug products were studied. The results indicated that the degradation products after combination tablets stored at room tempreture for one year were all from ketoconazole, which prompted that the degradation products of ketoconazole should be concerned in further research.On the basis of quality research of ketoconazole and beauvericin combination drug tablets, the quality standard was developed, include properties of appearance, identification, content determination, content uniformity, related substances, dissolution rate and frangibility.