Mps1-IN-2_Mps1激酶抑制剂_1228817-38-6_Apexbio
Mps1-IN-1_Mps1激酶抑制剂_1125593-20-5_Apexbio
产品描述:
MPS1-IN-1 是 MPS1（单极纺锤体 1）激酶的选择性抑制剂，IC50 值为 367 nM。 MPS1（单极纺锤体 1 激酶）是纺锤体组装检查点的重要组成部分。它有利于 C-MAD2（闭合 MAD2）构象异构体的形成和 MCC（有丝分裂检验点复合物）的组装，参与维持染色体稳定 性和肿瘤生长。
特别声明
产品仅用于研究，
不针对患者销售，望谅解。
每个产品具体的储存和使用信息显示在产品说明书中。ApexBio 产品在推荐的条件下是稳定 的。产品会根据不同的推荐温度进行运输。许多产品短期运输是稳定的，运输温度不同于长 期储存的温度。我们确保我们的产品是在保持试剂质量的条件下运输的。收到产品后，按照 产品说明书上的要求进行储存。
OC1CCN(C2=CC(OC)=C(NC3=NC4=C(C=CN4)C(NC5=C(S(=O)(C(C)C)=O )C=CC=C5)=C3)C=C2)CC1
>16.05mg/mL in DMSO
Store at -20°C
For obtaining a higher solubility , please warm the tube at 37°C and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20°C for several months.
MPS1-IN-1 废除 SAC（纺锤体装配检验点）的功能。在 U2OS 细胞中，在 20 分钟内，MPS1-IN-1 剂量依赖性的减少几乎 100％的细胞中有丝分裂花费的时间，开始进入有丝分裂后期。相比 之下，在同一时期内，DMSO 处理的细胞仅有 10％开始进入有丝分裂后期。在 MPS1-IN-1 处理的细胞中，加速有丝分裂动力学可影响基因组稳定性，引起非整倍体形成。此外， MPS1-IN-1 处理的细胞中结合动粒的 Mad2 降低 80％。相比于 DMSO 处理的细胞，MPS1-IN-1 处理的细胞中有丝分裂花费的时间减少大约 40％。此外，MPS1-IN-1 干扰 Aurora B 的激酶活 性。MPS1-IN-1 以剂量依赖性方式降低 Aurora B 激酶 Thr232 的磷酸化状态。此外，MPS1-IN-1 增加多极细胞分裂，降低细胞活力。
FDA批准的激酶小分子抑制剂类药物及分类一览
FDA批准的激酶小分子抑制剂类药物及分类一览蛋白激酶蛋白激酶(Kinase)是细胞生命活动重要的信号使者，可催化将ATP末端的γ-磷酸基团转移至底物上，从而将各种信号进行传递(图1)。
蛋白激酶参与了众多的生理过程，包括细胞增殖、存活、凋亡、代谢、转录以及分化等。
药理学及病理学研究表明，对于很多疾病，如肿瘤、炎症性疾病、中枢神经系统疾病、心血管疾病及糖尿病等，蛋白激酶都是一个理想的药物靶点。
图1 Mechanism of protein kinases and related publications对于蛋白激酶的研究始于20世纪50年代，并在90年代随着MAPK/ERK、JAK及PI3K等信号通路的揭示而达到一个研究热潮。
迄今为止，在人体中发现了518种蛋白激酶，而编码具有激酶活性蛋白的基因则高达900多种。
与之相对应，有关激酶抑制剂的研究也逐步发展，并在激酶作用机制的阐明过程中扮演了重要角色，并成为重要的药物研究热点。
该领域研究的文献数量也是逐年上升，从侧面反映了其在基础研究和药物发现中的重要性。
蛋白激酶抑制剂及其分类过去的15年间，激酶抑制剂作为药物候选的研究取得了长足的进步，不论是基础研究还是在工业界。
在人体现有药物靶点里面，蛋白激酶家族成员占比高达10%(FDA批准药物分子靶点深度解读)。
2001年，第一个激酶抑制剂类药物Imatinib获得FDA批准，成为该领域发展的里程碑，此后十年该类药物以平均每年获批一种的速度稳步发展。
而在2012年1月至2015年2月期间，小分子激酶抑制剂类药物迎来爆发式发展，共有15种新药获得审批。
截至2016年12月底，共有31种小分子激酶抑制剂类药物获得审批，同时还有大量的化合物处于临床或临床前研究中。
除此之外，科研人员还解析了超过5000种的蛋白激酶或蛋白激酶-抑制剂复合体的晶体结构，且超过五分之一的人类蛋白激酶具有明确的小分子抑制剂。
因此，小分子激酶抑制剂已成为药物研发的一个热点领域。
过氧化物酶体增殖物激活受体
过氧化物酶体增殖物激活受体(PPAR) 是一类由配体激活的核转录因子,属Ⅱ型核受体超家族成员, 存在3种亚型，即PPARα、PPARδ、PPARγ，这三种亚型在结构上有一定的相似性，均含DNA结合区和配体结合区等。
PPAR与配体结合后被激活，与9-顺视黄酸类受体形成异二聚体，然后与靶基因的启动子上游的过氧化物酶体增殖物反应元件(peroxisome proliferator response element，PPRE)结合而发挥转录调控作用。
PPRE 由含相隔一个或两个核苷酸的重复序列AGGTCA组成。
与配体结合后，PPAR在DNA结合区发生变构，进而影响PPAR刺激靶基因转录的能力。
PPARδ几乎在所有组织中表达，浓度低于PPARα及PPARγ，直至最近以前尚未找到此一核受体的选择性配基。
PPARδ是代谢综合征（肥胖、胰岛素抵抗、高血压是与脂质紊乱有关的共同的病态表现）的一个新靶点。
有不少的研究表明：GW501516可作为PPARδ的特异激动剂用于研究。
参考网址：/cjh/2003/shownews.asp?id=156/conference/preview.php?kind_id=03&cat_name=ADA2001&title_id=59219 Regulation of Muscle Fiber Type and Running Endurance by PPARδplos biology，Volume 2 | Issue 10 | October 2004/plosonline/?request=get-document&doi=10.1371%2Fjournal.pbio.0020294NF-KB通路中的抑制剂好像有1.PDTC(pyrrolidine dithiocarbamate),是一种抗氧化剂，主要作用于IκB降解的上游环节（IκBα的磷酸化或IKK的活性水平），2.Gliotoxin 是一种免疫抑制剂，机制可能从多个环节阻断NF-KB的激活，如IκB的降解，NF-KB的核移位和与DNA的结合。
FDA批准的激酶小分子抑制剂类药物及分类一览
FDA批准的激酶小分子抑制剂类药物及分类一览蛋白激酶蛋白激酶(Kinase)是细胞生命活动重要的信号使者，可催化将ATP末端的γ-磷酸基团转移至底物上，从而将各种信号进行传递(图1)。
蛋白激酶参与了众多的生理过程，包括细胞增殖、存活、凋亡、代谢、转录以及分化等。
药理学及病理学研究表明，对于很多疾病，如肿瘤、炎症性疾病、中枢神经系统疾病、心血管疾病及糖尿病等，蛋白激酶都是一个理想的药物靶点。
图1 Mechanism of protein kinases and related publications对于蛋白激酶的研究始于20世纪50年代，并在90年代随着MAPK/ERK、JAK 及PI3K等信号通路的揭示而达到一个研究热潮。
迄今为止，在人体中发现了518种蛋白激酶，而编码具有激酶活性蛋白的基因则高达900多种。
与之相对应，有关激酶抑制剂的研究也逐步发展，并在激酶作用机制的阐明过程中扮演了重要角色，并成为重要的药物研究热点。
该领域研究的文献数量也是逐年上升，从侧面反映了其在基础研究和药物发现中的重要性。
蛋白激酶抑制剂及其分类过去的15年间，激酶抑制剂作为药物候选的研究取得了长足的进步，不论是基础研究还是在工业界。
在人体现有药物靶点里面，蛋白激酶家族成员占比高达10%(FDA批准药物分子靶点深度解读)。
2001年，第一个激酶抑制剂类药物Imatinib获得FDA批准，成为该领域发展的里程碑，此后十年该类药物以平均每年获批一种的速度稳步发展。
而在2012年1月至2015年2月期间，小分子激酶抑制剂类药物迎来爆发式发展，共有15种新药获得审批。
截至2016年12月底，共有31种小分子激酶抑制剂类药物获得审批，同时还有大量的化合物处于临床或临床前研究中。
除此之外，科研人员还解析了超过5000种的蛋白激酶或蛋白激酶-抑制剂复合体的晶体结构，且超过五分之一的人类蛋白激酶具有明确的小分子抑制剂。
因此，小分子激酶抑制剂已成为药物研发的一个热点领域。
微管蛋白聚合抑制剂类抗肿瘤药Plinabulin
微管蛋白聚合抑制剂类抗肿瘤药Plinabulin
佚名
【期刊名称】《药学进展》
【年(卷),期】2011(35)4
【摘要】@@ 鉴于血管生成对肿瘤的生长和转移至关重要,因此肿瘤血管系统已成为极具价值的肿瘤治疗靶.目前,靶向肿瘤脉管系统的抗肿瘤药物主要包括抗血管生成剂和血管阻断剂(VDAs),前者可抑制肿瘤新血管的生长,而后者则是靶向破坏为肿瘤细胞供应氧和营养的已有血管网.肿瘤血管具有与正常血管不同的异常结构,其内皮细胞增殖迅速,血管壁变薄而具高渗透性,导致高血流阻力.
【总页数】2页(P185-186)
【正文语种】中文
【中图分类】R979.19
【相关文献】
1.美国授权酪氨酸激酶抑制剂类抗肿瘤药物的专利现状分析 [J], 张海龙
bretastatins类微管蛋白抑制剂的定量构效关系与结合模式 [J], 田然;季宇彬;刘振明;金宏威;张亮仁;林文翰
3.乙酰胆碱受体抑制剂对猪脑微管蛋白聚合的影响 [J], 宋忠魁;梁子卿;汪旭
4.微管蛋白聚合靶向抑制的抗肿瘤药的研究进展 [J], 许悦
5.几种小分子c-Met抑制剂类抗肿瘤药物概述 [J], 肖飞;彭红;金鑫;张婷
因版权原因，仅展示原文概要，查看原文内容请购买。
低毫安电化学疗法抑制磷酸肌醇3激酶蛋白激酶B信号通道抗人乳腺癌细胞株MDA-MB231侵袭迁移的机制重点
ＥＣＴ对细胞侵袭、迁移能力改变；用反转
ＥＣＴ作用ＭＤＡ－ＭＢ２３１细胞２４ ｈ后血管内
皮生长因子（ＶＥＧＦ）、基质金属蛋白酶（ＭＭＰ）－２、第１０号染色体上缺失与张力蛋白同源的磷酸酯酶 基因（ＰＴＥＮ）、蛋白激酶Ｂ（Ａｋｔ）基因ｍＲＮＡ和蛋白的表达。结果Ｔｒａｎｓｗｅｌｌ实验证实５ 后ＭＤＡ－ＭＢ２３１细胞侵袭、迁移能力下降；ＲＴ—ＰＣＲ结果显示５
ｓｏｍｅｔｅｎ（ＰＴＥＮ），Ａｋｔ ａｎｄ ｐｈｏｓｐｈｏｒｙｌａｔｅｄ Ａｋｔ（Ｐ—Ａｋｔ）ｉｎ ＭＤＡ—ＭＢ２３
ｔｉｖｅ
ｌ
ｃｅｌｌｓ．Ｍｅｔｈｏｄｓ
Ｔｒａｎｓｗｌｌ
ｓｈｏｗｅｄ ＥＣＴ ｃｏｕｌｄ ｉｎｈｉｂｉｔ ｔｈｅ ａｂｉｌｉｔｙ ｏｆ ｃｅｌｌ ｍｉｇｒａｔｉｏｎ ａｎｄ ｉｎｖａｓｉｏｎ ｏｂｖｉｏｕｓｌｙ．ＲＴ—ＰＣＲ ｓｈｏｗｅｄ ｔｈｅ ｒｅｌａ— ｅｘｐｒｅｓｓｉｏｎ ｌｅｖｅｌｓ ｏｆ ＶＥＧＦ ａｎｄ ＭＭＰ一２ ｉｎ ５ Ｃ ａｎｄ １０ Ｃ ＥＣＴ ｇｒｏｕｐｓ ｗｅｒｅ ｒｅｄｕｃｅｄ ｇｒａｄｕａｌｌｙ ｆ ｆｏｒ
ｓｈｅｎｇ，Ｚｈａｎｇ Ｚｈｉ，Ｚｈａｎｇ ｓＤ昭，Ｓｕ Ｚｈｅｎｍｉｎ，Ｓｈｅｎ ｒｉｊⅡｎ，Ｗａｎｇ Ｊｉａｎ Ｄｅｐａｒｔｍｅｎｔ ｏｆ Ｂｒｅａｓｔ Ｃａｎｃｅｒ Ｍｉｎｉｍａｌｌｙ Ｉｎｖａｓｉｖｅ Ｓｕｒｇｅｒｙ，Ｎｉｎｇｘｉａ Ａｕｔｏｎｏｍｏｕｓ Ｒｅｇｉｏｎ Ｐｅｏｐｌｅ’ｓ Ｈｏｓｐｉｔａｌ， Ｙｉｎｃｈｕａｎ ７５０００２，Ｃｈｉｎａ（Ｚｈｏｕ ＢＧ，Ｗａｎｇ Ｊ）；Ｄｅｐａｒｔｍｅｎｔ ｏｆ Ｂｒｅａｓｔ Ｃａｎｃｅｒ Ｓｕｒｇｅｒｙ，Ｇａｎｓｕ Ｐｒｏｖｉｎｃａｌ Ｃａｎｃｅｒ Ｈｏｓｐｉｔａｌ，Ｌａｎｚｈｏｕ ７３００５１９，Ｃｈｉｎａ（Ｗｅｉ ＣＳ）；Ｇｒａｄｕａｔｅ Ｃｏｌ如ｇｅ，Ｎｉｎｇｘｉａ Ｍｅｄｉｃａｌ Ｕｎｉｖｅｒｓｉｔｙ，Ｙｉｎ－ ｃｈｕａｎ ７５０００４，Ｃｈｉｎａ（Ｚｈａｎｇ Ｚ，Ｚｈａｎｇ Ｓ，Ｓｕ ＺＭ，Ｓｈｅｎ ＹＪ）
选择性 MPS1 抑制剂可影响胶质母细胞瘤的药物敏感性
DOI:10.1093/jnci/djt168Advance Access publication August 12, 2013© T he Author 2013. Published by Oxford University Press. All rights reserved.For Permissions, please e-mail: journals.permissions@.Articleeffects of the Selective MPS1 inhibitor MPS1-iN-3 on Glioblastoma Sensitivity to Antimitotic DrugsBakhos A. Tannous, Mariam Kerami, Petra M. Van der Stoop, Nicholas Kwiatkowski, Jinhua Wang, Wenjun Zhou, Almuth F . Kessler, Grant Lewandrowski, Lotte Hiddingh, Nik Sol, Tonny Lagerweij, Laurine Wedekind, Johanna M. Niers, Marco Barazas, R. Jonas A. Nilsson, Dirk Geerts, Philip C. De Witt Hamer, Carsten Hagemann, W. Peter Vandertop, Olaf Van Tellingen, David P . Noske, Nathanael S. Gray, Thomas Würdinger Manuscript received August 3, 2011; revised November 19, 2012; accepted May 8, 2013.Correspondence to: Thomas Würdinger, PhD, VU University Medical Center, CCA room 3.34, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands, (e-mail: t.wurdinger@vumc.nl ).B ackgroundGlioblastomas exhibit a high level of chemotherapeutic resistance, including to the antimitotic agents vincristineand taxol. During the mitotic agent-induced arrest, glioblastoma cells are able to perform damage-control and self-repair to continue proliferation. Monopolar spindle 1 (MPS1/TTK) is a checkpoint kinase and a gatekeeper of the mitotic arrest.MethodsWe used glioblastoma cells to determine the expression of MPS1 and to determine the effects of MPS1 inhibition on mitotic errors and cell viability in combination with vincristine and taxol. The effect of MPS1 inhibition was assessed in different orthotopic glioblastoma mouse models (n = 3–7 mice/group). MPS1 expression levels were examined in relation to patient survival.ResultsUsing publicly available gene expression data, we determined that MPS1 overexpression corresponds positively with tumor grade and negatively with patient survival (two-sided t test, P < .001). Patients with high MPS1 expres-sion (n = 203) had a median and mean survival of 487 and 913 days (95% confidence intervals [CI] = 751 to 1075), respectively, and a 2-year survival rate of 35%, whereas patients with intermediate MPS1 expression (n = 140) had a median and mean survival of 858 and 1183 days (95% CI = 1177 to 1189), respectively, and a 2-year survival rate of 56%. We demonstrate that MPS1 inhibition by RNAi results in sensitization to antimitotic agents. We developed a selective small-molecule inhibitor of MPS1, MPS1-IN-3, which caused mitotic aberrancies in glioblastoma cells and, in combination with vincristine, induced mitotic checkpoint override, increased aneuploidy, and augmented cell death. MPS1-IN-3 sensitizes glioblastoma cells to vincristine in orthotopic mouse models (two-sided log-rank test, P < .01), resulting in prolonged survival without toxicity.C onclusions Our results collectively demonstrate that MPS1, a putative therapeutic target in glioblastoma, can be selectively inhibited by MPS1-IN-3 sensitizing glioblastoma cells to antimitotic drugs.J Natl Cancer Inst;2013;105:1322–1331Glioblastoma, the highest grade glioma, is the most common and lethal type of primary brain tumor. Glioblastoma patients have a median survival of less than 15 months following standard of care (1). The main reason for this grim outcome is the rapid tumor growth and invasion of the surrounding brain parenchyma and the failure of standard radiotherapy and temozolomide chemotherapy and addi-tional treatments, such as the use of antimitotic agents, including vincristine and taxol (2,3). Recent advances in expression profil-ing technologies have allowed the exploratory analysis of differen-tial gene expression in an attempt to identify potential therapeutic targets for cancer therapy. We previously identified a set of kinases to be highly overexpressed in glioma. Apart from WEE1, CDK1, AURKA, and BUBR1, one of the top-rank overexpressed cell cycle-related kinases was MPS1, with an unclear role in glioma (4).Monopolar spindle 1 (MPS1, also known as TTK), is an evolutionary conserved dual specificity protein kinase that regulates the mitotic spindle checkpoint by monitoring proper chromosome attachment to spindle microtubules (5). As long as unattached kinetochores—the structure where the spindles attach to the chromosomes—are present, the mitotic checkpoint proteins will halt the cell cycle progress until all chromosomes are aligned and stably attached to the spindle. Upon stable orientation of chromosomes in metaphase, chromosome segregation is allowed to proceed (6). MPS1 exerts checkpoint control by redirecting several essential proteins to the kinetochores, including MAD1 and MAD2 (7,8). Furthermore, MPS1 regulates chromosome alignment during metaphase (8–12). Besides its checkpoint function, MPS1 has a probable role in centrosome duplication andat Huazhong University of Science and Technology on October 17, 2014/Downloaded fromin cytokinesis (5). I t is also reported to be involved in the p53-dependent postmitotic checkpoint (13), CHK2 signaling (14), and noncanonical Smad signaling by phosphorylation of Smad2 and Smad3 (15). Misregulation of MPS1 kinase activity results in chromosomal instability and, consequently, in aneuploidy (10). This is a common cause of tumor heterogeneity and poor prognosis in particular for patients with glioma (16–18).Antimitotic agents such as vincristine and taxol have been in clinical and oncological use for many years and cause mitotic arrest at the metaphase/anaphase boundary. This can result in a decrease in cancer cell proliferation and reduced tumor growth (19–21). However, many cancers, including gliomas, are resistant to these drugs (2,3,22,23). I t was previously demonstrated that targeting the mitotic checkpoint through inhibition of MPS1 can lead to accelerated mitosis and apoptosis in cancer cells with no effect on normal fibroblast cells (24). Simultaneous targeting of the mitotic checkpoint and chromosome alignment by inhibition of MPS1 in combination with low doses of the antimitotic drug taxol was shown to result in sensitization of HeLa, HCT-116, LS1740, and U2OS cells to taxol by elevating the frequency of chromosome missegregation (25).Several MPS1 inhibitors have been developed (24–30). However, compelling data to demonstrate their anticancer activity and safety have not been reported, and so far no MPS1 inhibitor has entered clinical testing in cancer patients. Here we describe the profile of a newly developed, selective, and highly potent MPS1 kinase inhibitor, MPS1-I N-3. We exploit the high expression of MPS1 in glioma and demonstrate that inhibition of this kinase by MPS1-IN-3 sensitizes glioblastoma cells to antimitotic agents in vitro and in vivo, providing a potential therapeutic strategy for the treatment of high-grade gliomas.MethodsPatient samplesFor the REMBRANDT cohort, samples were previously collected from the 14 contributing institutions (National Institutes of Health, Henry Ford Hospital, Thomas Jefferson University, University of California at San Francisco, H. Lee Moffitt Hospital, University of Wisconsin, University of Pittsburgh Medical Center, University of California at Los Angeles, The University of T exas M.D. Anderson Cancer Center, Dana-Farber Cancer Center, Duke University, Johns Hopkins University, Massachusetts General Hospital, and Memorial Sloan Kettering Cancer Center) (33). The samples were provided as snap-frozen sections of areas immediately adjacent to the region used for the histopathologic diagnosis. Initial histo-pathologic diagnosis was performed at the tissue-collecting insti-tution following World Health Organization (WHO) standards and reviewed by neuropathologists at a central laboratory (33). Additional data on the patient characteristics have been previously pubished (33). In addition, we used the publicly available data of the R2 French glioma cohort (R2.amc.nl) (43). These glioma samples were collected from the tumor archive of the Erasmus University Medical Center, Rotterdam, the Netherlands. All histologic diagno-ses were made on formalin-fixed, paraffin-embedded hematoxylin and eosin stained sections and were reviewed blinded to the original diagnosis according to the 2007 WHO classification (43).Statistical AnalysisThe median inhibition concentration (IC50) values were calculated using GraphPad Prism 5.0 (GraphPad Software, La Jolla, CA) using nonlinear curve fitting. Mice survival data were analyzed using the same software, and statistical significance was determined basedon log-rank test. Alternatively, the statistical s ignificance level (Student t test) was calculated using Microsoft Excel spreadsheets.A P value of less than .05 was considered statistically significant. All statistical tests were two-sided.Additional materials and methods are included in the Supplementary Methods (available online).resultsExpression Analysis of MPS1 as a Member of an Intricate Network of Mitotic Checkpoint GenesT o determine the expression status of spindle checkpoint genes (6) in glioma, we analyzed the expression of MPS1, BUBR1, AURKA, AURKB, BUB1, PLK1, MAD1, MAD2, and BUB3 using a publicly available dataset containing 152 glioma patientsand 23 control subjects (28). Hierarchical clustering analysis was performed to arrange samples and genes into groups based on their expression levels (Figure 1A). Three sample clusters were observed—one represented mainly by normal brain tissue, one bya mixture of glioma subtypes, and one by glioblastoma (Figure 1A).All spindle checkpoint genes analyzed, except for BUB3, were overexpressed in gliomasm, and their expression increased with tumor grade (Figure 1A). These results indicate that MPS1, the most differentially expressed of all mitotic spindle checkpoint genes analyzed, is part of a network of mitotic checkpoint genes that are collectively overexpressed in glioblastoma. T o further investigatethe correlation between MPS1 gene expression and tumor grade, publicly available gene expression data (28,31,32) were analyzed,and the P value for correlation between grade and MPS1 mRNA expression was determined (Figure 1B). MPS1 mRNA expression levels positively corresponded (P < .001, as provided by t test inthe Sun database accessed by Oncomine [http://www.oncomine. com]) to all tumor grades (ie, grade II: P = 1.22 × 10−6; grade III:P = 2.49 × 10−9; grade IV: P = 1.1 × 10−20, as provided by t test in the Sun database accessed by Oncomine). We confirmed the overexpressionof spindle checkpoint genes in a panel of three non-neoplastic brain (NNB) samples, 15 WHO grade II gliomas, and 15 WHO gradeI V gliomas by semiquantitative reverse-transcription polymerase chain reaction (Supplementary Figure 1, A–B, available online), aswell as in a panel of glioblastoma cell lines (data not shown). Next,we studied the correlation between MPS1 gene expression levelsand patient survival using the publicly available REMBRANDT data (33). As expected on the basis of tumor grade correlation, we found that high MPS1 expression statistically significantly and negatively correlated with survival of glioma patients (log rank P< .001) (Figure 1C). Patients with high MPS1 expression (≥3-fold increased MPS1 mRNA expression in glioma as compared withnon-neoplastic tissue expression; n = 203) had a median and mean survival of 487 and 913 days (95% CI = 751 to 1075), respectively,and a 2-year survival rate of 35%, whereas patients with intermediate MPS1 expression (<3-fold increased MPS1 mRNA expression in glioma as compared with non-neoplastic tissue expression; n = 140)at Huazhong University of Science and Technology on October 17, 2014/Downloaded fromFigure 1. Expression of MPS1 as a member of an intricate network of mitotic checkpoint proteins. A) Hierarchical clustering of samples was used to explore the similarities between expression profiles of mitotic spindle checkpoint proteins in glioma. Samples were divided into three clusters. T he bar graphs indicate the percentage samples within each cat-egory: astrocytoma grade II (A GII), grade III (A GIII), and grade IV (A GIV), oligodendroglioma grade I I (O GI I) and grade I I I (O GI I I), and control. The P values indicate the statistical significance of differential expres-sion as calculated using two-sided t test. B) MPS1 expression positively correlates with glioma grade (Oncomine database). C) Overexpression of MPS1 mRNA is associated with decreased survival in adult glioma patients. Kaplan–Meier plots derived from the publicly available REMBRANDT glioma gene expression dataset are grouped according to MPS1 mRNA expression level. Black curve represents high MPS1 expression: threefold or greater increase in MPS1 expression compared with non-neoplastic brain samples. Gray curve represents intermediate MPS1 expression (less than threefold increase in MPS1 mRNA). Patients with high MPS1 expression (threefold or greater increased MPS1 mRNA expression in glioma as compared with non-neoplastic tissue expres-sion; n=203) had a median and mean survival of 487 and 913 days (95% confidence interval [CI] = 751 to 1075), respectively, and a 2-year survivalrate of 35%, whereas the patients with intermediate MPS1 expression(less than threefold increased MPS1 mRNA expression in glioma as compared with non-neoplastic tissue expression; n=140) had a longer median and mean survival of 858 and 1183 days (95% CI = 1177 to 1189), respectively, and a higher 2-year survival rate of 56%. The number of patients at risk at 0, 1000, 2000, 3000, 4000, and 5000 days survival were respectively 203, 52, 23, 14, 7, and 3 for high MPS1 expression and 140,62, 28, 9, 4, and 1 for intermediate MPS1 expression. D) MPS1 protein expression was analyzed by staining of human glioma tissue as well as different regions of normal brain (cortex, ventricle, and hippocampus)as controls, provided by the Human Protein Atlas. Scale bar = 50 µm. E) Western blot analysis for MPS1 expression using extracts from glioblas-toma cell lines, primary cells, and glioblastoma tumor specimens.at Huazhong University of Science and Technology on October 17, 2014/Downloaded fromhad a longer median and mean survival of 858 and 1183 days (95% CI = 1177 to 1189), respectively, and a higher 2-year survival rate of 56% (Figure 1C). These results are explained by the correlation of MPS1 to the different glioma grades (Figure 1A). Similar results were obtained with an independent glioma dataset, which also show the effects of MPS1 expression on the survival within the different glioma subtypes, using the R2 French glioma cohort (R2.amc.nl) (43) (Supplementary Figure 1C, available online). T o study MPS1 overexpression at the protein level, we explored the publicly available Human Protein Atlas (http://www.proteinatlas. org). A robust MPS1 protein overexpression was observed in glioblastoma samples as compared with NNB (Figure 1D). T o confirm the MPS1 protein expression in glioma samples, we analyzed tissue lysates from NNB and glioblastoma tumor samples by Western blotting. MPS1 protein expression was detected in all glioblastoma samples, whereas no expression could be detected in NNB (Figure 1E). MPS1 protein expression was also confirmed in primary glioblastoma cell cultures and glioblastoma cell lines (Figure 1E).Effects of Inhibition of MPS1 by RNA Interference on Mitotic Aberrancies and Glioblastoma Cell Proliferation Previously, it was demonstrated that knockdown of MPS1 causes mitotic abnormalities, confirming its role in normal mitotic pro-gression (34). We thus decided to investigate whether MPS1 knockdown causes abnormal mitosis in glioblastoma cell cultures. We used the H2B-GFP fusion reporter to monitor mitotic aber-rancies. U251 glioblastoma cells expressing firefly luciferase (Fluc), and the mCherry fluorescent protein and H2B-GFP (U251-FM-H2B-GFP) were transduced with lentivirus vectors encod-ing a control shRNA (shCTRL) or a previously validated shRNA (Sigma) directed against MPS1 (shMPS1). Western blot analysis showed efficient MPS1 knockdown in cells expressing shMPS1 as compared with shCTRL (Figure 2A). Fluorescent microscopy showed mitotic errors as indicated by abnormal H2B-GFP locali-zation (Figure 2B). These abnormalities ranged from lobed nuclei and bi- or multinucleated cells caused by defects in cytokinesis to anaphase bridges, chromosome blebs and strings, micronuclei, and lagging chromosomes during anaphase, representing chromosome attachment and segregation defects.Because MPS1 knockdown resulted in loss of control of the mitotic checkpoint, we determined whether MPS1 knockdown in combination with antimitotic agents could cause a chromosome missegregation phenotype. We therefore treated U251-FM-H2B-GFP cells expressing shCTRL or shMPS1 with the antimitotic agent vincristine or taxol. T o investigate the effect of MPS1 knock-down on vincristine-induced cell death, we incubated U251 glio-blastoma cells with low doses (3 nM) of vincristine and assessed cell growth after 11 days. Vincristine treatment statistically significantly reduced the number of viable U251-shMPS1 cells as compared with U251-shCTRL cells (t test, P < .01) (Figure 2C). T reatment with taxol in combination with MPS1 knockdown demonstrated similar effects as vincristine (Figure 2D). T o assess a chromosome missegregation phenotype, U251-FM-H2B-GFP cells express-ing shCTRL or shMPS1 and treated with low doses of vincristine (3 nM) or taxol (5 nM) for 7 days were analyzed for nuclear mor-phology by fluorescence microscopy. As expected, drastic nuclear aberrancies, ranging from lobed nuclei and multinucleated cells suggesting cytokinesis defects to the presence of chromosome bridges and micronuclei, which reflect gross chromosome segrega-tion defects, were observed in shMPS1 cells that had divided in the presence of vincristine or taxol (Figure 2, E and F). T o investigatethe consequence of MPS1 knockdown on tumor growth in vivo, an orthotopic mouse model was employed. U251-FM cells expressing shCTRL or shMPS1 were used, and tumor growth was monitoredby Fluc bioluminescence imaging. MPS1 knockdown resulted ina modest delay of tumor progression (Figure 2G; Supplementary Figure 2A, available online). We then determined whether MPS1 knockdown could sensitize glioblastoma tumors to vincristine-mediated cell death in vivo. U251-FM cells expressing shCTRL or shMPS1 were injected into the brain of mice, and 10 days later, mice were treated with intravenous injections of vincristine (0.5 mg/kg) once per week for 2 weeks. This relatively low dose of vincristine caused a modest reduction of tumor growth. On the other hand, inhibition of MPS1 by shMPS1 caused cessation of tumor growth, demonstrating nearly complete sensitization to vincristine (t test,P = .003), as measured by bioluminescence imaging at 3 weeks after implantation of the cells (Figure 2H; Supplementary Figure 2B, available online). Collectively, these data demonstrate that MPS1 knockdown in combination with low doses of antimitotic agents causes lethal chromosome segregation errors, ultimately leading to glioblastoma cell death. These results are consistent with previous findings that connect MPS1-mediated taxol sensitization to chro-mosome segregation errors (25).Development of the Selective and Potent MPS1 InhibitorMPS1-IN-3Previously, MPS1 inhibitors were identified from an in vitro ATP-site competition-binding assay by screening a diverse library of het-erocyclic ATP-site directed kinase scaffolds (26). These inhibitors, MPS1-IN-1 and MPS1-IN-2, represented distinct chemical scaf-folds; however, both displayed potent activity against MPS1. In the course of our medicinal chemistry efforts, we discovered a closely related analog of MPS1-IN-1, MPS1-IN-3, which potently inhib-ited MPS1 kinase activity with an IC50 of 50 nM (SupplementaryT able 1, available online). T o arrive at MPS1-IN-3, the pyrollopyri-dine core of MPS1-IN-1 was replaced with a purine core scaffold (Figure 3A; Supplementary Methods, available online). This chem-ical modification resulted in nearly a fivefold increase in activity against MPS1 in a biochemical assay (Supplementary Figure 3, available online). T o test whether MPS1-IN-3 abrogates the spin-dle checkpoint, we challenged U2OS cells arrested in mitosis with increasing concentration of MPS1-IN-3. Silencing of the spindle checkpoint, a surrogate readout of MPS1 activity, was assessed by monitoring the stability of cyclin B (Figure 3B). The levels of cyclinB, a marker of mitotic cells, dropped with increasing concentra-tion of MPS1-IN-3 but was reversed by addition of the proteasome inhibitor MG132 (Figure 3B). Complete checkpoint abrogation was observed with 2 μM of MPS1-IN-3. Downregulation of this mitotic marker indicated that MPS1-IN-3 caused a dose-depend-ent escape from a checkpoint-mediated mitotic arrest. I n addi-tion, we confirmed functional MPS1 kinase inhibition in U251 glioblastoma cells by analyzing the phosphorylation levels of the MPS1 target Smad2 by Western blotting using MPS1-I N-3 atat Huazhong University of Science and Technology on October 17, 2014/Downloaded fromFigure 2. The effect of inhibition of MPS1 by RNA interference on mitotic aberrancies and glioblastoma cell proliferation. A) Western blot analysis showing MPS1 knockdown in U251 cells using shMPS1. B) U251-shCTRL and U251-shMPS1 cells expressing H2B-GFP were analyzed for changes in nuclear morphology using fluorescence microscopy. Scale bar = 10 µm.C and D) U251 cells expressing shMPS1 or shCTRL were treated with low concentrations of vincristine (3 nM) or taxol (5 nM), and 11 days after addition, cell viability was assessed using the crystal violet assay. Data shown as average ± standard deviation. **P < .01, two-sided t test. E) U251-FM-H2B-GFP cells expressing shCTRL or shMPS1 were treatedwith 3 nM of vincristine or 5 nM of taxol for 7 days, after which cells were analyzed using fluorescence microscopy for nuclear morphology. Scalebar = 10 µm. F) Quantitation of nuclear aberrancies visualized in (E). Datashown as average ± standard deviation. ***P < .001, two-sided t test. G)Nude mice (n = 3) were injected intracranially with U251-FM-shCTRL or-shMPS1 cells. Tumor growth was analyzed by firefly luciferase biolumi-nescence imaging at week 7. H) Similar group of mice (n = 3) as in (G) treated with 0.5 mg/kg of vincristine (Vinc) or DMSO (CTRL).at Huazhong University of Science and Technology on October 17, 2014/Downloaded from5 µM (Figure 3C ), and MPS1-I N-3 inhibited the proliferation of U251 glioblastoma cells as determined by cell counting with an I C 50 value of approximately 5 µM (Figure 3D ), demonstrat-ing improved effects over the inhibitors MPS-IN-2 and AZ-3146 (Supplementary Figure 3, available online). I n addition, U251 cells were subjected to pMPM2 fluorescence activated cell sortinganalysis in the presence of vincristine, demonstrating a reduction inthe number of U251 cells in vincristine-induced mitotic arrest after MPS1-IN-3 treatment (Figure 3E ). T o determine whether MPS1 inhibition by MPS1-IN-3 in combination with antimitotic agents causes chromosome missegregation phenotypes, we treated U251-FM-H2B-GFP cells with MPS1-IN-3 at 5 µM and low doses ofFigure 3. Development of the selective and potent MPS1 inhibi-tor MPS1-N-3. A ) Chemical development of MPS1-N-3. Compound 1—2,6-dichloro-9H -purine—was first converted by reaction a to com-pound 2—2,6-dichloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purine—then by reaction b to compound 3—2-chloro-N-(2-(isopropylthio)phenyl)-9-(tetrahydro-2H -pyran-2-yl)-9H -purin-6-amine—and reaction c to product 4—2-chloro-N-(2-(isopropylsulfonyl)phenyl)-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine—to finally reach desired product 5 by reaction d—1-(4-(6-(2-(isopropylsulfonyl)phenylamino)-9H-purin-2-ylamino)-3-ethoxyphenyl)piperidin-4-ol (MPS1-I N-3). Reactions, rea-gents, and conditions are described in the Supplementary Experimental Procedures (available online). B ) Western blot analysis of cyclin B in U2OS cells treated with nocodazole, MPS1-I N-3, and/or MG132. C )Western blot analysis of Smad2 phosphorylation in U251 cells incubated with 5 μM of MPS1-IN-3 (MPS1i) and vincristine. D ) Dose–response curveof MPS1-IN-3 on U251 glioblastoma cells as analyzed by cell counts. Data shown as average ± standard deviation. E ) Fluorescence activated cellsorting analysis of pMPM2 in U251 cells 24 hour after addition of 5 μM of MPS1-IN-3 (MPS1i) and vincristine. Percentages indicate cells in mitoticarrest. F ) U251-FM-H2B-GFP cells were treated with DMSO (CTRL) or MPS1-IN-3 (MPS1i) in combination with vincristine or taxol. The nuclearmorphology was assessed with fluorescence microscopy and quantitatedin (G ). Scale bar = 10 µm. The experiment was performed in triplicate. Shown are mean averages, and error bars indicate standard deviation.at Huazhong University of Science and Technology on October 17, 2014/Downloaded fromvincristine (3 nM) or taxol (5 nM), and after 1 week, cells were ana-lyzed for nuclear morphology by fluorescence microscopy. Similar to MPS1 knockdown, drastic nuclear aberrancies, including lobed nuclei and multinucleated cells, and micronuclei reflecting gross chromosome segregation defects were observed in U251 cells treated with MPS1-I N-3 in the presence of vincristine or taxol (Figure 3, F and G). In conclusion, MPS1-IN-3 is a selective and potent MPS1 inhibitor with phenotypic consequences similar to those reported for published MPS1 inhibitors such as MPS1-IN-1, MPS1-IN-2, and AZD3146 (26,29,30).Effects of Inhibition of MPS1 by MPS1-IN-3 on Sensitivityof Glioblastoma Cells to Vincristine In Vitro and In Vivo Next, we determined the effect of MPS1-IN-3 on U251 and U87 glioblastoma cell lines and VU147 primary glioblastoma cells as well as GBM8 primary neurospheres in the presence or absence of vincristine. T reatment with MPS1-IN-3 at 5 µM sensitized all glioblastoma cells to 3 nM of vincristine as measured by cell counts 11 days posttreatment (Figure 4A). T o investigate the consequence of MPS1-IN-3 in combination with vincristine on tumor growth in vivo, an orthotopic glioblastoma mouse model was employed using U251-FM cells. U251-FM cells were stereotactically injected into the brain of nude mice. T en days postinjection, 2 mg/kg of MPS1-I N-3 and/or 0.5 mg/kg of vincristine were administered concomitantly by intravenous injections twice per week for 3 weeks. T umor growth was monitored by Fluc imaging, and mice were monitored for survival and for any signs of treatment tox-icity. A statistically significant decrease in Fluc bioluminescence signal and therefore tumor volume and an increase in survival were observed in mice treated with MPS1-I N-3 in combination with vincristine as compared with no treatment, in contrast with mice treated with MPS1-IN-3 or vincristine alone (t test, P < .05) (Figure 4, B–D). T o validate these results in a more clinically rel-evant model, we employed the orthotopic primary GBM8-FM neurosphere model, which infiltrates the mouse brain in a process similar to that in human GBM patients (35) using the same treat-ment conditions. A statistically significant decrease in biolumines-cence signal and increased survival were observed in mice treated with MPS1-IN-3 in combination with vincristine, in contrast with mice treated with vincristine or MPS1-I N-3 alone or with the no treatment controls (t test, P < .05) (Figure 4, E–G). An initial increase in Fluc signal was observed in the first week of vincristine monotreatment in the U251 model. However, we did not observe this phenomenon in the GBM8 model. The differences in growth inhibition of the U251 and GBM8 models we attribute to the dif-ferent nature of the U251 GBM cell line and the primary GBM8 stemlike cells. We noticed no toxicity upon treatment of the mice, as was confirmed by additional toxicity analysis. C57BL/6-naive mice were injected with either vehicle, vincristine, MPS1-IN-3, or a combination of vincristine and MPS1-N3 (similar doses to what we used for tumor treatment in vivo) on day 1 and day 3. On day 4, blood was collected and analyzed for complete blood count, a com-prehensive chemistry panel, and a liver enzyme panel, revealing no signs of toxicity (data not shown). Altogether these results demon-strate that MPS1-IN-3 sensitizes glioblastoma cells to vincristine both in vitro and in murine tumor models.DiscussionMost high-grade gliomas are resistant to conventional and experi-mental therapies including antimitotic agents such as taxol and vincristine (2,3). The MPS1 kinase controlled mitotic checkpointis an essential factor in determining the response of tumor cells to antimitotic drugs (36–39). Here, we demonstrate that MPS1 is a highly expressed mitotic checkpoint protein in glioma and a poten-tial therapeutic target in high-grade gliomas. We designed a new selective MPS1 inhibitor that effectively sensitizes glioblastomas to vincristine in two orthotopic glioblastoma xenograft models.Limitations of this study include the fact that MPS1-IN-3 was tested in a limited number of glioblastoma cell lines in combina-tions with vincristine. It would be of interest to further characterizethe effect of MPS1 inhibition on a vast number of primary glioblas-toma cell lines and to determine whether certain cellular expres-sion patterns correspond to the effectiveness of the drug. The same accounts for the in vivo studies, in which the U251 glioblastomacell line and the GBM8 primary glioblastoma stemlike cell modelswere used. I n addition, although effects of statistical significancewere measured in vivo, the exact pharmacodynamics and kinetics ofMPS1-IN-3 remain to be investigated. This includes studying thehalf-life of the drug, crossing of the blood brain barrier, drug pump substrate specificity, and functional target engagement (eg, by pSmad2 analysis) in different dosing schemes in combination with vincristine. T umor heterogeneity may play a role in the treatment response to MPS1-IN-3, and it would be of interest to study the effects of MPS1-IN-3 on different samples of the same tumor. Sofar, it is unclear whether resistance to MPS1 inhibition can occur in glioblastoma cells. Studies performing long-term exposure of glio-blastoma cells to MPS1-IN-3 and vincristine could reveal whethersuch mechanisms occur and, possibly, how this could be over-come. Finally, we observed minimal toxicity in our in vivo studies. However, we only studied toxicity in two different glioblastomain vivo models. Additional dose-escalating toxicity studies are war-ranted to optimize the combination of MPS1-IN-3 in combinationwith vincristine.Vincristine has been used in different combination regiments (procarbazine, lomustine, and vincristine) for the therapy ofhigh-grade oligodendroglioma and anaplastic oligodendroglioma (40,41). I ndeed, a recent meta-analysis emphasized the role of vincristine in high-grade glioma therapy by showing that patient cohorts treated with vincristine-containing regimens had statisti-cally significant survival advantage over cohorts treated with other chemotherapy drugs (42). However, the efficacy of vincristine in glioma treatment has been hampered by toxicity such as polyneu-ropathy and hematological side effects. Combination of vincristinewith MPS1 inhibitors may not only potentiate the antitumor effectof vincristine but may also ameliorate the notorious side effectsby allowing reduced vincristine dosing, although treatment ofboth normal and cancer cells with MPS1 inhibitors causes severe chromosome segregation defects and aneuploidy, suggesting thatthe tolerability of chronic administration of such agents warrants further investigation. In conclusion, our results demonstrate thatMPS1 may be a promising therapeutic target for high-grade gli-oma therapy and that MPS1 inhibition by MPS1-IN-3 efficiently sensitizes glioblastoma cells to antimitotic agents.at Huazhong University of Science and Technology on October 17, 2014/Downloaded from。
一种用于人磷酸化血管扩张刺激蛋白的酶联免疫检测试剂盒及其检测方法[发明专利]
![一种用于人磷酸化血管扩张刺激蛋白的酶联免疫检测试剂盒及其检测方法[发明专利]](https://img.taocdn.com/s3/m/b7e8120c2bf90242a8956bec0975f46526d3a759.png)
专利名称：一种用于人磷酸化血管扩张刺激蛋白的酶联免疫检测试剂盒及其检测方法
专利类型：发明专利
发明人：刘丹,易倩春
申请号：CN202111638413.9
申请日：20211230
公开号：CN114002440B
公开日：
20220401
专利内容由知识产权出版社提供
摘要：本发明公开了一种用于人磷酸化血管扩张刺激蛋白的酶联免疫检测试剂盒及其检测方法，其中，所述检测试剂盒包括样本前处理缓冲液、含PGE1的样本激活剂和含PGE1和ADP的样本抑制剂、酶标板、VASP单克隆抗体、显色液、终止液；其中VASP单克隆抗体为辣根过氧化物酶标记的鼠单抗，鼠单抗重链互补决定区包含CDR‑H1、CDR‑H2和CDR‑H3片段，鼠单抗轻链互补决定区包含CDR‑L1、CDR‑L2和CDR‑L3片段。
本发明采用酶联免疫法，并且对于单抗的序列进行了优化和改进，改进后的单克隆抗体捕获率高，特异性好，克服了全血样本对于检测结果的影响，免去了以往检测前还需要对待测样本本身进行处理的步骤。
申请人：湖南菲思特精准医疗科技有限公司
地址：410000 湖南省长沙市开福区沙坪街道中青路1048号山河医药健康产业园标准厂房3栋301-1
国籍：CN
代理机构：湖南乔熹知识产权代理事务所(普通合伙)
代理人：安曼
更多信息请下载全文后查看。
Pim—1激酶小分子抑制剂研究的现状和问题_1
Pim—1激酶小分子抑制剂研究的现状和问题转《上海医药》总结大全/html/zongjie/原癌基因pim-1是基因pim家族（现发现的有pim-1、pim-2、pim-3）中的一员，最早是作为莫洛尼小鼠白血病病毒（MoMuLV）的前病毒插入点被发现[1]，其编码的Pim-1激酶属于钙/钙调蛋白调节激酶（calcium/calmodulin-regulated kinase，CAMK），在多细胞组织的进化过程中高度保守。
pim-2和pim-3是pim家族的另外两个成员，他们与pim-1分别具有55%和65%的同源性。
pim-1在调控细胞凋亡，分化，增殖以及肿瘤形成等方面发挥了非常重要的作用。
Pim-1激酶能通过一个保守的甘氨酸环状基序磷酸化众多的在细胞凋亡、细胞周期调控中起重要作用的细胞因子，包括c-Myc，BAD，Socs1，Cdc25A，HP1，PAP-1，p21cip1/waf1，PTP-U2S，NFATc1等，从而导致肿瘤发生[2]。
Pim-1对FLT3-ITD AML具有上调作用，其抑制剂AR00459339可以促进FLT3下游靶点STAT5，AKT，BAD 的去磷酸化，对FLT3-ITD细胞如MV-4-11，Molm-14，和TF/ITD细胞系以及FLT3-ITD主要样品具有细胞毒性[3]。
Pim-1通过直接磷酸化Cdc25C推进细胞周期G2/M的进程，促进细胞的增殖分化[4]。
Pim-1还可通过与Socs1和Socs3抑制因子联合作用发挥其潜在的STAT5转录抑制作用。
Pim-1能够与NFATcl转录因子结合并使其丝氨酸片段磷酸化，促进IL-2的生成，启动IL-2依赖的淋巴细胞的增殖和成熟[5]。
最近研究表明Pim-1激酶在免疫调节、炎症（炎性肠疾病）以及新陈代谢和细胞增长中起着一定的作用[6-7]。
通过抑制Pim-1激酶，还可以增强Runx3的表达和阻止TH2及TH17 T细胞变异，从而预防花生过敏[8]。
对抗三阴乳腺癌化疗耐药性，MPS1靶点新药“重振”紫杉醇功效
对抗三阴乳腺癌化疗耐药性，MPS1靶点新药“重振”紫杉醇功效在乳腺癌里，有一种类叫做“三阴性乳腺癌”，尽管它大概只占乳腺癌总数的10%，但其造成的死亡病例，却占到了乳腺癌患者死亡数的1/4！由于该肿瘤细胞缺乏雌激素受体（ER）、孕激素受体（PR）和人表皮生长因子受体2（Her2），其治疗一直缺乏有效的靶向药物来延缓其患者的无进展生存期。
目前，靶向微管的药物（如紫杉醇）尽管存在副作用，但仍被视为对抗三重阴性乳腺癌的标准疗法。
但不可忽视的是，某些癌细胞会狡猾地“躲过一劫”，并导致患者对治疗产生抗药性。
Spiros Linardopoulos教师图片来源：ICR来自伦敦癌症研究所（ICR）的癌症生物学和治疗学教授Spiros Linardopoulos及其同事也关注到了这一临床现象，他们渴望找到一种方法来阻止这类“越狱”事件发生。
目前，其团队最新结果已发表在《Molecular Cancer Therapeutics》杂志上。
DOI: 10.1158/1535-7163.MCT-18-1203之前的研究发现，癌细胞因自行设定增殖速度而变得强大，可一旦它们分裂速度过快，则会在细胞增殖过程中导致DNA复制错误，尤其是染色体数量出现错误，而这些错误最终将导致癌细胞“自杀”。
研究人员猜想，既然了解了癌细胞“自杀”的发生机制，那是不是可以依此来进行药物治疗呢？BOS172722（DB15498）的结构研究人员关注了一款来自Boston Pharmaceuticals（波士顿制药公司）名为BOS172722的Mps1激酶抑制剂药物，该药物可迫使癌细胞的加速分裂以导致细胞凋亡。
其关注点在于：含单极纺锤体1（Mps1，也称为TTK）在内的多种有丝分裂激酶维持了染色体的稳定性，对染色体对齐和纺锤体装配检查点（SAC）起到至关重要的作用。
而BOS172722可通过靶向MPS1迫使细胞加速有丝分裂，导致染色体错位和错聚，终将引发细胞凋亡，对化疗后产生耐药性的癌症患者存在积极意义。
激酶抑制剂的新机制[英]
![激酶抑制剂的新机制[英]](https://img.taocdn.com/s3/m/ca5a251feffdc8d376eeaeaad1f34693daef10f8.png)
激酶抑制剂的新机制[英]王前（摘）;王林（摘）【期刊名称】《国外医学：药学分册》【年(卷),期】2006(33)4【摘要】大多已用于临床试验的激酶抑制剂，都是在激酶ATP结合位点，直接与ATP竞争发挥作用，例如成功的抗癌药伊马替尼（imatinib）即如此。
近期Adrian等报道了一类新型化合物，它与伊马替尼一样均抑制激酶BCR-ABL，不同的是它并非为ATP竞争机制，而是以一种新颖的别构效应起作用。
而且，这类化合物能减弱BCR-ABL突变体对伊马替尼的耐药性。
研究者对5000个化合物进行初筛后，对细胞毒性具有良好差异性的化合物作进一步研究，舍弃以ATP结合位点为靶点的化合物，【总页数】2页(P314-315)【关键词】激酶抑制剂;BCR-ABL;类化合物;伊马替尼;结合位点;竞争机制;ATP;临床试验;细胞毒性;抗癌药【作者】王前（摘）;王林（摘）【作者单位】【正文语种】中文【中图分类】R392.12;R733.7【相关文献】1.蛋白激酶2抑制剂增强盐酸埃克替尼对表皮生长因子受体－酪氨酸激酶抑制剂耐药细胞的增殖抑制作用及机制 [J], 周瑜;伍钢;孟睿;张盛;李珂;李倩雯;周方正;李振宇;马虹;董晓荣;刘莉2.蛋白激酶2抑制剂克服表皮生长因子受体-酪氨酸激酶抑制剂耐药的有效机制[J], 周瑜;张盛;李珂;董晓荣;刘莉;伍钢;孟睿3.酪氨酸蛋白激酶抑制剂Tyrphostin AG114对重组人蛋白激酶CK2全酶的抑制作用(英) [J], 刘新光;梁念慈4.在肿瘤细胞模型中联合应用磷脂酰肌醇3激酶/蛋白酶B通路抑制剂BEZ235和细胞外调解蛋白激酶/丝裂原活化蛋白激酶通路抑制剂U0126的效果 [J], 陈欣欣;张舒;石玉镯5.一种细胞外信号调节激酶和人乳腺癌细胞生长的新天然抑制剂[英]／Izevbigie E B…∥Exp Biol Med [J], 许重远;貌家安因版权原因，仅展示原文概要，查看原文内容请购买。
- 1、下载文档前请自行甄别文档内容的完整性，平台不提供额外的编辑、内容补充、找答案等附加服务。
- 2、"仅部分预览"的文档,不可在线预览部分如存在完整性等问题,可反馈申请退款(可完整预览的文档不适用该条件!)。
- 3、如文档侵犯您的权益，请联系客服反馈,我们会尽快为您处理(人工客服工作时间：9:00-18:30)。
参考文献: [1] Kwiatkowski N1, Jelluma N, Filippakopoulos P, Soundararajan M, Manak MS, Kwon M, Choi HG, Sim T, Deveraux QL, Rottmann S, Pellman D, Shah JV, Kops GJ, Knapp S, Gray NS. Small-molecule kinase inhibitors provide insight into Mps1 cell cycle function. Nat Chem Biol. 2010 May;6(5):359-68. doi: 10.1038/nchembio.345. Epub 2010 Apr 11.
产品说明书
化学性质
产品名: Mps1-IN-2 修订日期: 6/30/2016
产品名: Cas No.: 分子量: 分子式: 化学名:
SMILES:
溶解性: 储存条件: 一般建议:
运输条件:
Mps1-IN-2
1228817-38-6
480.6
C26H36N6O3
9-cyclopentyl-2-[2-ethoxy-4-(4-hydroxypiperidin-1-yl)anilino]-5-met hyl-7,8-dihydropyrimido[4,5-b][1,4]diazepin-6-one
特别声明
产品仅用于研究， 不针对患者销售，望谅解。 每个产品具体的储存和使用信息显示在产品说明书中。ApexBio 产品在推荐的条件下是稳定 的。产品会根据不同的推荐温度进行运输。许多产品短期运输是稳定的，运输温度不同于长 期储存的温度。我们确保我们的产品是在保持试剂质量的条件下运输的。收到产品后，按照 产品说明书上的要求进行储存。
产品描述:
Mps1-IN-2 是一种有效的\选择性的和 ATP 竞争性的 Mps1 激酶抑制剂,其 IC50 值为 145 nM[1]. 据报道,在 1 μM ATP 存在时,Mps1-IN-2 可以抑制 Mps1 激酶活性,其 IC50 值为 145 nM.此 外,Mps1-IN-2 对 Mps1 的选择性比对 Alk 和 Ltk 酶高 1000 多倍.Mps1-IN-2 结合到 Mpa1 的 ATP 结合口袋,在 2.74 分辨率下,观察到 Mps1-IN-2 与该激酶铰链区(Glu603)形成氢键.Mps1-IN-2 通 过直接抑制 Mps1,从而废除纺锤体组装检查点(SAC)功能[1].
Evaluation sample solution : ship with blue ice All other available size: ship with RT , or blue ice upon request
生物活性
靶点 :
Cell Cycle/Checkpoint
信号通路:
Mps1
ApexBio Technology
CCOC1=C(C=CC(=C1)N2CCC(CC2)O)NC3=NC=C4C(=N3)N(CCC(=O)N4 C)C5CCCC5
Soluble in DMSO
Store at -20°C
For obtaining a higher solubility , please warm the tube at 37°C and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20°C for several months.