Zornes L L, Stratford R E
Department of Pharmaceutical Sciences, Lilly Research Laboratories, Lilly Corporate Center, Indianapolis, IN 46285, USA.
J Chromatogr B Biomed Sci Appl. 1997 Aug 1;695(2):381-7. doi: 10.1016/s0378-4347(97)00184-9.
An HPLC assay for plasma analysis of LY303366 (I), a semi-synthetic lipopeptide antifungal related to echinocandin B (ECB), was developed to support the selection and subsequent preclinical development of I. The method involved extraction of I from plasma with the aid of solid-phase extraction (SPE) cartidges followed by reversed-phase HPLC with UV detection at 300 nm. The method is simple, selective and is applicable to dog, rat, mouse and rabbit plasma. Validation studies using dog plasma showed that the values obtained for parameters of linearity, precision and accuracy were within acceptable limits. Based on analysis of 0.3 ml of plasma, the lower limit of quantitation was 20 ng/ml. The method has been successfully applied to determine the pharmacokinetic parameters of I in the dog following intravenous (i.v.) and oral administration. Compared to first generation ECB antifungal agents, the results of the i.v. dog study indicated a 50% reduction in clearance of the drug from plasma (0.1 l/h/kg) and an 18-fold increase in the volume of distribution at steady state (1.8 l/kg). When administered orally, compound I had an absolute bioavailability of 9%; however, plasma levels remained above the MIC for C. albicans (0.005 microg/ml) through 48 h. Given the excellent potency of I and its broad spectrum of activity relative to first generation ECB antifungal agents, the assay results for I indicate the potential for its use as a broad spectrum i.v. and oral antifungal agent.
开发了一种用于血浆分析LY303366（I）的高效液相色谱（HPLC）测定法，LY303366是一种与棘白菌素B（ECB）相关的半合成脂肽抗真菌剂，以支持I的筛选及后续临床前开发。该方法包括借助固相萃取（SPE）柱从血浆中提取I，然后进行反相HPLC，并在300 nm处进行紫外检测。该方法简单、具有选择性，适用于犬、大鼠、小鼠和兔的血浆。使用犬血浆进行的验证研究表明，线性、精密度和准确度参数所获得的值在可接受范围内。基于对0.3 ml血浆的分析，定量下限为20 ng/ml。该方法已成功应用于测定犬静脉注射（i.v.）和口服给药后I的药代动力学参数。与第一代ECB抗真菌剂相比，犬静脉注射研究结果表明，该药物从血浆中的清除率降低了50%（0.1 l/h/kg），稳态分布容积增加了18倍（1.8 l/kg）。口服给药时，化合物I的绝对生物利用度为9%；然而，在48小时内血浆水平仍高于白色念珠菌的最低抑菌浓度（MIC，0.005 μg/ml）。鉴于I相对于第一代ECB抗真菌剂具有优异的效力及其广泛的活性谱，I的测定结果表明其有潜力作为一种广谱静脉注射和口服抗真菌剂使用。
