肿瘤血管抑制剂Combretastatins的重要中间体CA4酸质量研究

Comprehensive study on the quality of CA4 acid as the important intermediate of tumor vascular inhibitor Combretastatins

对Combretastatin系列化合物的重要中间体CA4酸(CA4s)理化特征、有关物质、残留溶剂和含量测定的方法进行系统研究。通过IR、UV、MS、NMR等确定本品的化学结构。采用HPLC法检测有关物质,条件为:色谱柱C18(150 mm×4.60 mm,5μm),流动相(体积比):0.1%甲酸(氨水调解pH至6.8)-甲醇-乙腈(75∶22∶3),检测波长为220 nm;采用HS-GC法检测残留溶剂,条件为:PEG20M(30 m×0.53 mm,1μm)的石英毛细管柱,柱温80℃,检测器温度250℃(FID),进样口温度200℃,流量4.0 mL/min,分流比:10∶1;采用LC-MS鉴定本品有关物质;以剩余滴定法测定本品的含量。结果表明:CA4酸为难溶于水的浅黄色结晶性粉末,其化学结构与2-(3,4,5-三甲氧苯基)-3-[3-羟基-4-甲氧苯基]-丙烯酸相吻合,有6个杂质组分,经鉴定杂质5和杂质2为三甲氧基苯乙酸和异香草醛;杂质6为Z式CA4酸,受光照影响发生较大的变化反映出顺反异构的特征;残留溶剂乙醇量小于0.08%。CA4酸为Combretastatinl类化合物的关键中间体,难溶于水,分子中的羧基可与碱性化合物作用生成可溶性的盐,可突破该类化合物难溶性瓶颈,提高该类化合物抑制肿瘤血管的效率,是一个具有较高价值Combretastatins中间体或先导化合物。

To set up a systematic study, including the analysis on the physical and chemical features and the establishment of the methods for analysis of related substances, determination of content and the determination of residual solvent, on the CA4 acid-an important intermediates of Combretastatins, a tumor angiogenesis inhibitors -and provide the foundation for farther study on this kind of angiogenesis inhibitors. The chemical structure was confirmed by IR , UV, MS and NMR. Related substances was analyzed by HPLC and the chromatographic conditions were as follows： C18（150mm×4.60mm,5 μ m）; the mobile phase contained a mixture of 0.1% formic acid buffer （ammonia to pH 6. 8）, methanol and acetonitrile in a ratio of 75：22：3 （v/ v/ v ）; with the wavelongth of 220 nm; The HS -GC chromatographic conditions for determination of residual solvent ethanol were as follows： analyzing in PEG capillary column （30m×0.53mm, 1μ m） with column temperature at 80℃ ,the detector temperature of FID at 250℃ and injection port temperature at 200 ℃,the flow rate of carrier gas nitrogen was 4.0 mL/min with diversion ratio of 10：1. Related substances were identified by LC/MS, and the content of CA4 acid was determined by the volumetric analysis. CA4 acid was light yellow amorphous powder, and was almost insoluble in water. The structure of CA4 acid is （E）-3-[（3-hydroxy-4-methoxyphenyl）]-2- （3, 4, 5 -trimethoxyphenyl） acrylic acid. The fifth and second of its爷 six related substances were trimethoxy phenylacetic acid and isovanillin, the starting materials of CA4 acid, while the sixth was the （Z） -CA4 acid, which transformed from CA4 acid mainly because of light influences. The content of residual solvent ethanol was less than 0.08%. As the carboxyl group in the molecule can react with an alkaline compound to form a soluble salt, CA4 acid is seen as the important intermediates of Combretastatins which could enhance the efficiency of inhibiting tumor blood vessels.