Olaparib_COA_14425_MedChemExpress

奥拉帕尼（Olaparib）合成检索总结报告
一、奥拉帕尼（Olaparib）简介
奥拉帕尼（Olaparib）于2014年12月19日在美国上市。

奥拉帕尼（Olaparib）是聚腺苷酸二磷酸核糖转移酶抑制剂，用于晚期卵巢癌。

奥拉帕尼（Olaparib）不良反应有：恶心、疲乏、呕吐、腹泻、味觉障碍、消化不良等等。

奥拉帕尼（Olaparib）分子结构式如下:
英文名称：Olaparib
中文名称：奥拉帕尼
本文主要对奥拉帕尼（Olaparib）的合成路线、关键中间体的合成方法及实验操作方法进行了文献检索并作出了总结。

二、奥拉帕尼（Olaparib）合成路线一
三、奥拉帕尼（Olaparib）合成路线二
四、奥拉帕尼（Olaparib）合成路线一检索总结报告
(一) 奥拉帕尼（Olaparib）中间体3的合成方法一（路线一）
(二) 奥拉帕尼（Olaparib）中间体3的合成方法二（路线一）
(三) 奥拉帕尼（Olaparib）中间体3的合成方法三（路线一）①奥拉帕尼（Olaparib）中间体8的合成。

体外肝代谢系统【摘要】肝药酶在药物代谢中具有十分重要的作用。

对肝药酶的研究方法中，以动物肝脏或肝细胞为基础，构建体外肝代谢系统是体外代谢研究中最重要的环节之一。

对体外肝代谢的研究，主要是利用肝微粒体、基因重组CYP450酶系、肝细胞培养、肝组织切片及离体肝灌流系统等方法。

本文综述近年国内外所应用的不同体外肝代谢系统，并对各体外代谢研究方法进行比较，指出根据各系统的特性、不同的实验要求和目的，选择适当的研究方法的重要性。

【关键词】细胞色素P450酶；肝微粒体；肝细胞培养；肝组织切片；离体肝灌流药物代谢一般是指药物的生物转化。

药物经生物转化后，可引起药物的药理活性或∕和毒理活性的改变。

因此，研究药物的生物转化，明确其代谢过程，对新药开发、新剂型设计及制定合理的临床用药方案等方面都具有重要的指导意义。

肝脏是药物生物转化的重要器官，含有参与药物代谢重要的酶系组成，主要有CYP1、CYP2、CYP3三大家族［1］。

本文所介绍的各种体外代谢系统均含有一种或多种CYP450酶的同工酶，为研究药物体外代谢提供了研究的对象和基础。

动物肝体外代谢研究可以较好地排除体内因素干扰，直接观察酶对底物代谢的选择性，为整体试验提供可靠的科学依据。

以肝脏为基础的体外代谢系统主要包括肝微粒体、基因重组CYP450酶系、肝细胞、肝组织切片及离体肝灌流。

1肝微粒体肝微粒体的制备多数采用差速离心法［2］，通过高速离心使微粒体与其他成分分离，操作简单，无需其他试剂辅助。

但较耗时，设备要求高，使该法的普及和深入研究受到一定的限制。

针对这些情况，可采用试剂辅助分离的方法［3］，在离心前额外加入一定比例的PEG6000或CaCl2，促进微粒体沉降。

此法对设备要求降低，并缩短了实验周期。

肝微粒体的制备过程均应在4℃下进行。

正确、合理地选择缓冲液，能起到良好介质的作用，按比例加入后进行肝组织的破碎和匀浆，才可有效分离肝微粒体和避免细胞器受损。

肝微粒体的主要应用测定CYP450酶活性测定原理是在特定酶催化下，底物在辅助因子以及适合的温度、时间作用下反应，借助仪器测定生成的特定产物量。

不可切除胰腺癌的分子靶向药物治疗进展胡润，李俊蒽，姚沛，桂仁捷，段华新湖南师范大学附属第一医院，湖南省人民医院肿瘤科，长沙 410005通信作者：段华新，****************（ORCID： 0000-0001-9596-5013）摘要：胰腺癌作为消化系统最常见的恶性肿瘤之一，其发病率及死亡率正逐年上升，大多数胰腺癌患者因分期较晚而失去了手术机会。

尽管以吉西他滨、氟尿嘧啶为主的化疗方案在一定程度上延长了患者的生存期，但仍有部分患者因无法耐受化疗而失去治疗机会。

随着精准医疗时代的来临，分子靶向药物治疗展现出的优异疗效使其成为对抗肿瘤的重要治疗手段之一，但由于胰腺癌高度的异质性及复杂的免疫微环境，针对胰腺癌的分子靶向治疗并未取得显著效果，因此亟需探寻新的治疗靶点及药物攻克这一难题。

本综述基于胰腺癌常见分子靶点及肿瘤免疫相关靶点探究在不可切除胰腺癌中分子靶向药物治疗研究的最新进展，为胰腺癌患者提供新的治疗策略。

关键词：胰腺肿瘤；分子靶向治疗；免疫疗法基金项目：湖南省自然科学基金（2020JJ8084）Advances in molecular-targeted therapy for unresectable pancreatic cancerHU Run，LI Junen，YAO Pei，GUI Renjie，DUAN Huaxin.（Department of Oncology，The First Affiliated Hospital of Hunan Normal University， Hunan Provincial People’s Hospital， Changsha 410005， China）Corresponding author： DUAN Huaxin，****************（ORCID： 0000-0001-9596-5013）Abstract：Pancreatic cancer is one of the most prevalent malignant tumors of the digestive system， and its incidence and mortality rates are increasing year by year. Most patients with pancreatic cancer are unable to receive surgery due to the advanced stage. Although chemotherapy regimens based on gemcitabine and fluorouracil have prolonged the survival time of patients to some extent，some patients cannot tolerate chemotherapy and hence lose the opportunity for treatment. With the advent of the era of precision medicine， molecular-targeted therapy has exhibited an excellent therapeutic efficacy and has thus become one of the most important treatment techniques for tumors； however， due to the high heterogeneity of pancreatic cancer and its complicated tumor microenvironment， molecular-targeted therapy for pancreatic cancer has not achieved notable results. Therefore， it is imperative to seek new therapeutic targets and medications to overcome this issue. This article reviews the latest advances in the research on molecular-targeted therapy for unresectable pancreatic cancer based on common molecular targets and tumor immunity-related therapeutic targets， in order to provide new treatment strategies for patients with pancreatic cancer.Key words：Pancreatic Neoplasms； Molecular Targeted Therapy； ImmunotherapyResearch funding：Natural Science Foundation of Hunan Province of China （2020JJ8084）胰腺癌是一种起病隐匿、进展迅速、疗效及预后极差的恶性肿瘤，大多数患者确诊时已经属于晚期。

Inhibitors, Agonists, Screening Libraries Data SheetBIOLOGICAL ACTIVITY:CHIR–99021 is a GSK–3α/β inhibitor with IC 50 of 10 nM/6.7 nM; > 500–fold selectivity for GSK–3 versus its closest homologs CDC2 and ERK2, as well as other protein kinases.IC50 & Target: IC50: 10 nM/6.7 nM (GSK–3α/β)[1]In Vitro: CHIR 99021inhibits human GSK–3β with K i values of 9.8 nM [1]. CHIR 99021 is a small organic molecule that inhibits GSK3α and GSK3β by competing for their ATP–binding sites.In vitro kinase assays reveal that CHIR 99021 specifically inhibits GSK3β (IC 50=~5 nM) and GSK3α (IC 50=~10 nM), with little effect on other kinases [2]. In the presence of CHIR–99021 the viability of the ES–D3 cells is reduced by 24.7% at 2.5 μM, 56.3% at 5 μM, 61.9% at 7.5 μM and 69.2% at 10 μM CHIR–99021 with an IC 50 of 4.9μM [3].In Vivo: In ZDF rats, a single oral dose of CHIR 99021 (16 mg/kg or 48 mg/kg) rapidly lowers plasma glucose, with a maximal reduction of nearly 150 mg/dl 3–4 h after administration [1]. CHIR99021 (2 mg/kg) given once, 4 h before irradiation, significantly improves survival after 14.5 Gy abdominal irradiation (ABI). CHIR99021 treatment significantly blocks crypt apoptosis andaccumulation of p–H2AX + cells, and improves crypt regeneration and villus height. CHIR99021 treatment increases Lgr5+ cellsurvival by blocking apoptosis, and effectively prevents the reduction of Olfm4, Lgr5 and CD44 as early as 4 h [4].PROTOCOL (Extracted from published papers and Only for reference)Kinase Assay:[2]Kinases are purified from SF9 cells through use of their His or Glu tag. Glu–tagged proteins are purified, and His–tagged proteins are purified. Kinase assays are performed in 96–well plates with appropriate peptide substrates in a 300–μL reaction buffer (variations on 50 mM Tris–HCl, pH 7.5, 10 mM MgCl 2, 1 mM EGTA, 1 mMdithiothreitol, 25 mMβ–glycerophosphate, 1mM NaF, and 0.01% bovine serum albumin). Peptides has K m values from 1 to 100 μM. CHIR 99021 or CHIR GSKIA is added in 3.5μL of Me 2SO, followed by ATP to a final concentration of 1 μM. After incubation, triplicate 100–μL aliquots are transferred to Combiplate 8 plates containing 100 μL/well of 50 μM ATP and 20 mM EDTA. After 1 hour, the wells are rinsed five times with phosphate–buffered saline, filled with 200 μL of scintillation fluid, sealed, and counted in a scintillation counter 30 min later. All of the steps are at room temperature. The percentage of inhibition is calculated as 100×(inhibitor–no enzyme control)/(Me 2SO control–no enzyme control)[2].Cell Assay: CHIR 99021 is dissolved in DMSO and stored, and then diluted with appropriate media before use [3].[3]The viability of the mouse ES cells is determined after exposure to different concentrations of GSK3 inhibitors for three days using the MTT assay.The decrease of MTT activity is a reliable metabolism–based test for quantifying cell viability; this decrease correlates with the loss of cell viability. 2,000 cells are seeded overnight on gelatine–coated 96–well plates in LIF–containing ES cell medium. On the next day the medium is changed to medium devoid of LIF and with reduced serum and supplemented with 0.1–1 μM BIO, or 1–10 μM SB–216763, CHIR–99021 or CHIR–98014. Basal medium without GSK3 inhibitors or DMSO is used as control. All tested conditions are analyzed in triplicates [3].Product Name:CHIR–99021Cat. No.:HY-10182CAS No.:252917-06-9Molecular Formula:C 22H 18Cl 2N 8Molecular Weight:465.34Target:GSK–3; GSK–3; Autophagy Pathway:Stem Cell/Wnt; PI3K/Akt/mTOR; Autophagy Solubility:DMSO: ≥ 5.1 mg/mLAnimal Administration: CHIR 99021 is formulated as solutions in 20 mM citrate–buffered 15% Captisol or as fine suspensions in0.5% carboxymethylcellulose (Rat)[1].CHIR 99021 is prepared in DMSO and diluted (Mice)[4].[1][4]Rat[1]Primary hepatocytes from male Sprague Dawley rats that weighed <140 g are prepared and used 1–3 h after isolation. Aliquotsof 1×106cells in 1 mL of DMEM/F12 medium plus 0.2% BSA and CHIR 99021(orally at 16 or 48 mg/kg) or controls are incubated in 12–well plates on a low–speed shaker for 30 min at 37°C in a CO2–enriched atmosphere, collected by centrifugation and lysed by freeze/thaw in buffer A plus 0.01% NP40; the GS assay is again performed.Mice[4]Mice 6–10 weeks old are used. The PUMA+/+ and PUMA–/– littermates on C57BL/6 background (F10) and Lgr5–EGFP(Lgr5–EGFP–IRES–creERT2) mice are subjected to whole body irradiation (TBI), or abdominal irradiation (ABI). Mice are injected intraperitoneally (i.p.) with 2 mg/kg of CHIR99021 4 h before radiation or 1 mg/kg of SB415286 28 h and 4 h before radiation. Mice are sacrificed to collect small intestines for histology analysis and western blotting. All mice are injected i.p. with 100 mg/kg of BrdU before sacrifice.References:[1]. Ring DB, et al. Selective glycogen synthase kinase 3 inhibitors potentiate insulin activation of glucose transport and utilization in vitro and in vivo. Diabetes. 2003 Mar;52(3):588–95.[2]. Bennett CN, et al. Regulation of Wnt signaling during adipogenesis. J Biol Chem. 2002 Aug 23;277(34):30998–1004.[3]. Naujok O, et al. Cytotoxicity and activation of the Wnt/beta–catenin pathway in mouse embryonic stem cells treated with four GSK3 inhibitors.BMC Res Notes. 2014 Apr 29;7:273.[4]. Wang X, et al. Pharmacologically blocking p53–dependent apoptosis protects intestinal stem cells and mice from radiation. Sci Rep. 2015 Apr 10;5:8566.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。