摘 要：迄今为止，肺癌已然成为全球死亡率极高的恶性肿瘤之一，严重威害着人们的身心健康。为此医学界、制药界等相关科研人士积极研制出了一种能够提高肺癌临床治疗效果，可以延长肺癌患者的生存期限的药物——克唑替尼(Crizotinib)。此药的发明和应用，给个体化治疗带去了希望和突破，是抗癌新药开发的一个重大里程碑。在制备克唑替尼中，手性中间体(S)-1-(2,6-二氯-3-氟苯基)乙醇是其合成的关键。而合成关键中间体则经历了从步骤繁琐，收率较低的手性拆分法到高转化率但反应条件苛刻、成本昂贵的化学合成法再到绿色但不易回收的酶催化法，从传统的低产率的生物催化技术到可循环利用但酶易失活的固定化酶技术再到而今风靡的生物酶催化，科学界正在一步步探索着更简洁高效的合成克唑替尼关键手性中间体的路线。因此，本课题主要研究一条新型路线，以2,6-二氯-3-氟苯乙酮为原料，以酮还原酶和脱氢酶的共固定为基础，形成一种以磷酸钙为载体的酶-无机杂化纳米花，并在固定化系统中双酶的协同催化下制备2,6-二氯-3-氟苯乙醇这关键手性中间体，以及在实验中探究出此酶-无机杂化纳米花的性能和优势所在。
实验证明：1.制备所得的酶晶体的热稳定性明显高于游离酶的热稳定性，60℃下8个小时酶晶体中的酶仍能保持80%以上的活性，而游离酶只保持20%；2.酶晶体的催化活性强于游离酶，且最佳催化条件40℃，；3.制备所得的酶晶体有效地保证了16个催化循环中基本上所有的催化活性，大大提高了酶的重复利用率，节约了成本。
关键词：肺癌； 手性中间体； 共固定化； 酶-无机杂化纳米花； 协同催化
Abstract：So far, lung cancer has become one of the most serious malignant tumors in the world. It has seriously harmed people's physical and mental health. So, the related scientific research personnel have actively developed crizotinib that can increase the clinical treatment effect of lung cancer and can prolong the survival period of patients. The invention and application of this drug brought hope and breakthrough to inpidualized treatment. It is a major milestone in the development of new anti-cancer drugs. In the preparation of crizotinib, the chiral intermediate (S)-1-(2,6-dichloro-3-fluorophenyl)ethanol is the key to its synthesis. The synthesis of key intermediates has gone from the tedious, low-yield chiral resolution to high conversion but harsh reaction conditions, the costly chemical synthesis to the green but difficult to recover enzyme catalysis, from the traditional, low-yield biocatalytic technology to the reusable but inactivated the immobilized enzyme technology to the now-popular bio-enzyme catalysis. The scientific community is exploring the route to a more compact and highly efficient synthesis of the key chiral intermediates of crizotinib. Therefore, this project mainly studies a new route which using 2,6-dichloro-3-fluoroacetophenone as a raw material, based on the co-immobilization of ketoreductase and dehydrogenase, to form a lipase-inorganic hybrid nanoflower with calcium phosphate. And the synergistic catalysis of two enzymes in the immobilization system to prepare 2,6-dichloro-3-fluorophenylethanol which called a key chiral intermediate, and to explore the performance and advantages of the lipase-inorganic hybrid nanoflower.
The experimental results show that：1. The thermal stability of the prepared enzyme crystals is obviously higher than the thermal stability of the free enzyme, and the enzyme in the crystals of the enzyme can still maintain 80% or more of activity for 8 hours at 60°C，while the free enzyme only maintains 20%；2. The catalytic activity of the enzyme crystals is stronger than the catalytic activity of the free enzyme, and the optimal catalytic conditions is 40°C；3. The prepared enzyme crystals effectively ensure substantially all the catalytic activities in the 16 catalytic cycles，greatly increasing the reuse of the enzyme and saving the cost. 酮还原酶与脱氢酶的共固定与性能研究:http://www.chuibin.com/huaxue/lunwen_206012.html