Selective type II TRK inhibitors overcome xDFG mutation mediated acquired resistance to the second-generation inhibitors selitrectinib and repotrectinib

Neurotrophic receptor kinase(NTRK) fusions are actionable oncogenic drivers of multiple pediatric and adult solid tumors,and tropomyosin receptor kinase(TRK) has been considered as an attractive therapeutic target for "pan-cancer" harboring these fusions.Currently,two generations TRK inhibitors have been developed.The representative second-generation inhibitors selitrectinib and repotrectinib were designed to overcome clinic acquired resistance of the first-generation inhibitors larotrectinib or entrectinib resulted from solvent-front and gatekeeper on-target mutations.However,xDFG(TRKAG667C/A/S,homologous TRKCG696C/A/S) and some double mutations still confer resistance to selitrectinib and repotrectinib,and overcoming these resistances represents a major unmet clinical need.In this review,we summarize the acquired resistance mechanism of the first-and second-generation TRK inhibitors,and firstly put forward the emerging selective type Ⅱ TRK inhibitors to overcome xDFG mutations mediated resistance.Additionally,we concluded our perspectives on new challenges and future directions in this field.

Discovery of a highly selective VEGFR2 kinase inhibitor CHMFL-VEGFR2-002 as a novel antiangiogenesis agent

Angiogenesis is an essential process in tumor growth,invasion and metastasis.VEGF receptor 2(VEGFR2)inhibitors targeting tumor angiogenic pathway have been widely used in the clinical cancer treatment.However,most of currently used VEGFR2 kinase inhibitors are multi-target inhibitors which might result in target-associated side effects and therefore limited clinical toleration.Highly selective VEGFR inhibitors are still highly demanded from both basic research and clinical application point of view.Here we report the discovery and characterization of a novel VEGFR2 inhibitor(CHMFLVEGFR2-002),which exhibited high selectivity among structurally closed kinases including PDGFRs,FGFRs,CSF1 R,etc.CHMFL-VEGFR2-002 displayed potent inhibitory activity against VEGFR2 kinase in the biochemical assay(IC50=66 nmol/L)and VEGFR2 autophosphorylation in cells(EC50s^100 nmol/L)as well as potent anti-proliferation effect against VEGFR2 transformed BaF3 cells(GI50=150 nmol/L).In addition,CHMFL-VEGFR2-002 also displayed good anti-angiogenesis efficacy in vitro and exhibited good in vivo PK(pharmacokinetics)profile with bioavailability over 49%and antiangiogenesis efficacy in both zebrafish and mouse models without apparent toxicity.These results suggest that CHMFL-VEGFR2-002 might be a useful research tool for dissecting new functions of VEGFR2 kinase as well as a potential anti-angiogenetic agent for the cancer therapy.

Development of small molecule extracellular signal-regulated kinases(ERKs) inhibitors for cancer therapy

The mitogen-activated protein kinase(MAPK)/extracellular signal-regulated kinase 1/2(ERK1/2) signaling pathway is widely activated by a variety of extracellular stimuli, and its dysregulation is associated with the proliferation, invasion, and migration of cancer cells. ERK1/2 is located at the distal end of this pathway and rarely undergoes mutations, making it an attractive target for anticancer drug development. Currently, an increasing number of ERK1/2 inhibitors have been designed and synthesized for antitumor therapy, among which representative compounds have entered clinical trials. When ERK1/2 signal transduction is eliminated, ERK5 may provide a bypass route to rescue proliferation, and weaken the potency of ERK1/2 inhibitors. Therefore, drug research targeting ERK5 or based on the compensatory mechanism of ERK5 for ERK1/2 opens up a new way for oncotherapy. This review provides an overview of the physiological and biological functions of ERKs, focuses on the structure-activity relationships of small molecule inhibitors targeting ERKs, with a view to providing guidance for future drug design and optimization, and discusses the potential therapeutic strategies to overcome drug resistance.

Class I phosphatidylinositol 3-kinase inhibitors for cancer therapy

The phosphatidylinositol 3-kinase(PI3K) pathway is frequently activated in human cancers.Class I PI3 Ks are lipid kinases that phosphorylate phosphatidylinositol 4,5-bisphosphate(PIP2) at the 3-OH of the inositol ring to generate phosphatidylinositol 3,4,5-trisphosphate(PIP3), which in turn activates Akt and the downstream effectors like mammalian target of rapamycin(m TOR) to play key roles in carcinogenesis. Therefore, PI3 K has become an important anticancer drug target, and currently there is very high interest in the pharmaceutical development of PI3 K inhibitors. Idelalisib has been approved in USA and Europe as the first-in-class PI3 K inhibitor for cancer therapy. Dozens of other PI3 K inhibitors including BKM120 and ZSTK474 are being evaluated in clinical trials. Multifaceted studies on these PI3 K inhibitors are being performed, such as single and combinational efficacy, resistance, biomarkers,etc. This review provides an introduction to PI3 K and summarizes key advances in the development of PI3 K inhibitors.

选择性PI3K抑制剂的研究进展

磷脂酰肌醇3-激酶(PI3K)是细胞内重要的信号转导分子,在细胞存活、增殖和分化过程中起重要调节作用。PI3K是PI3K/AKT/mT0R信号转导通路中的关键节点蛋白,调控着重要的生命活动。现已证实磷脂酰肌醇3-激酶(PI3Ks)是潜力巨大的药物治疗靶点,特别是PI3Kα现己成为抗肿瘤治疗的重要靶点之一。目前己有多种针对PI3Ks的抑制剂进入临床研究。然而现有抑制剂的化学类型不多且选择性不高、临床应用受局限。因此积极研究和开发结构新颖的PI3K选择性抑制剂对于疾病的靶向治疗具有重要意义。本文将对选择性PI3K抑制剂在抗肿瘤方面的研究进展作一综述。

Rho激酶对结直肠癌细胞体外侵袭转移作用的研究

目的:观察Rho激酶对结直肠癌SW480细胞黏附、运动、迁移和侵袭能力的影响。方法:SW480细胞用Rho激酶抑制剂Y-27632处理后,分别用细胞运动实验,黏附实验和迁移、侵袭实验分析对肿瘤细胞运动、黏附、迁移和侵袭能力的影响。结果:25μmol/L以上的浓度的Y-27632作用于SW480细胞后,细胞的运动能力、迁移能力明显下降(p<0.05),细胞黏附于纤维粘连蛋白的能力也降低(p<0.05);而且细胞穿过人工基底膜的能力明显降低(p<0.01)。结论:Rho激酶参与结直肠癌细胞体外的侵袭转移作用,Rho激酶抑制剂有望成为有效的抗恶性肿瘤转移的药物。

小分子选择性表皮生长因子受体抑制剂的研究进展

表皮生长因子受体(EGFR)是一种具有蛋白酪氨酸激酶(PTKs)活性的跨膜受体。EGFR及其配体是细胞信号传导系统的一部分,与肿瘤的形成和恶化密切相关,是治疗肿瘤的一个很有前景的靶点。EGFR抑制剂可以阻断EGFR酪氨酸激酶的传导通路,抑制肿瘤细胞的增殖、分化和迁移。本文就2000-2016年间不同类型的选择性EGFR抑制剂的研究进展作一综述。

复发/难治DLBCL患者继发CNS侵犯的临床特征及泽布替尼联合化疗方案的疗效

目的分析4例继发中枢神经系统(CNS)侵犯的复发/难治弥漫大B细胞淋巴瘤(DLBCL)患者的临床特征,并观察泽布替尼联合化疗方案的疗效。方法选择4例DLBCL继发CNS侵犯患者,分析其初诊及复发时的临床特征;并进行相应泽布替尼联合化疗方案治疗,其中,2例采用泽布替尼+甲氨蝶呤(Z-MTX)方案治疗,1例采用泽布替尼+利妥昔单抗+吉西他滨+顺铂+地塞米松(Z-R-GDP)方案治疗,1例采用泽布替尼+利妥昔单抗+甲氨蝶呤(ZR-MTX)方案治疗,评价治疗方案的疗效及毒性反应。结果4例患者中,3例CNS复发仅侵犯脑实质,另1例同时侵犯脑、眼;3例具有淋巴瘤中枢侵袭的高危中枢神经系统国际预后指数(CNS-IPI)评分(4~6分),另1例CNS-IPI评分1分。3例患者为非生发中心(non-GCB)来源,其中MCD亚型2例、非MCD亚型1例;1例未进行相关分子生物学检查。化疗后完全缓解3例,部分缓解1例;随访4~5个月,至末次随访,3例完全缓解患者疗效持续缓解无复发,1例部分缓解患者复发。主要毒性反应为血液学毒性,未发生肿瘤溶解、心房颤动或高血压等其他3级及以上布鲁顿氏酪氨酸激酶选择性抑制剂相关的毒性反应。结论复发/难治DLBCL继发CNS侵犯多为non-GCB来源及MCD亚型,主要侵犯脑实质;泽布替尼联合化疗方案有显著、持续的疗效和生存优势,其主要毒性反应可控。

毛细管电泳-激光诱导荧光检测用于多个胞内蛋白激酶的抑制剂筛选和选择性评价(英文)

发展了一种基于毛细管电泳(CE)-激光诱导荧光(LIF)检测的多个细胞内源激酶的抑制剂平行筛选及选择性评价方法。CE高效的分离能力和LIF检测器的高选择性,使得同时测试多个胞内激酶的活性成为可能。共4种细胞系、3种特异性蛋白激酶底物肽、2种选择性蛋白激酶抑制剂和1种非选择性蛋白激酶抑制剂用于方法的建立。特异性底物肽与细胞裂解液混合后孵育,被其相应的激酶选择性地磷酸化,利用CE-LIF分离检测磷酸化产物和底物肽。同时测定一个抑制剂对几种蛋白激酶的抑制活性,用于评价抑制剂的选择性。与传统的单靶标筛选模式相比,这种基于细胞裂解液的多靶标筛选方法能提供更多的信息,更加高效,且细胞裂解液作为一种廉价的激酶来源大大降低了筛选成本。

二苯基嘧啶类化合物抗癌活性的研究进展

二苯基嘧啶类化合物(DPPYs)因其特有的骨架结构能与癌细胞中的多种靶点结合,其中很多化合物还具有活性高、选择性好和抗突变株能力强等优点,多个化合物处于临床研究中,代表性药物奥希替尼已于2015年被美国食品药品监督管理局(FDA)批准上市且很快成为"重磅炸弹药物"。根据不同的靶点将该类化合物分为六类,综述了国内外DPPYs的抗癌活性研究进展。对于具有多靶点活性的部分化合物,将其归类到靶点研究最为深入的抑制剂中。DPPYs在抗癌药物研究中前景广阔,值得关注。

4-芳胺基喹唑啉类化合物在激酶靶向小分子抑制剂中的应用

恶性肿瘤已经成为我国最严重的疾病之一。蛋白激酶和肿瘤密切相关,选择性的抑制特定的激酶,可以起到靶向治疗癌症的效果。4-芳胺基喹唑啉类化合物是很重要的一类激酶靶向小分子抑制剂。介绍了4-芳胺基喹唑啉类化合物在激酶靶向小分子抑制剂中的应用及其普遍合成方法。

Janus激酶2选择性抑制剂HipHop药效团模型与3D QSAR模型的建立

目的为Janus激酶2(JAK2)选择性抑制剂的设计提供指导。方法将从已报道文献中筛选出的27个JAK2选择性抑制剂小分子通过Discovery Studido 4.5软件的分子共同特征药效团(HipHop)定性算法与定量构效关系(QSAR)分别构建JAK2选择性抑制剂的模型。结果HipHop药效团模型具有较强的筛选能力,且可以预测化合物是否具有活性;3D QSAR模型不仅可以预测化合物的活性值,还对化合物的设计提供指导;二者结合应用更加有助于发现新型JAK2选择性抑制剂。结论HipHop药效团和3D QSAR模型方法可以寻找可能的抗癌药物JAK2抑制剂先导分子,为后续的研究奠定理论基础。

Synthesis of selective PAK4 inhibitors for lung metastasis of lung cancer and melanoma cells

The p21 activated kinase 4(PAK4) is serine/threonine protein kinase that is critical for cancer progression.Guided by X-ray crystallography and structure-based optimization,we report a novel subseries of C-3-substituted 6-ethynyl-1 H-indole derivatives that display high potential and specificity towards group Ⅱ PAKs.Among these inhibitors,compound 55 exhibited excellent inhibitory activity and kinase selectivity,displayed superior anti-migratory and anti-invasive properties against the lung cancer cell line A549 and the melanoma cell line B16.Compound 55 exhibited potent in vivo antitumor metastatic efficacy,with over 80% and 90% inhibition of lung metastasis in A549 or B16-BL6 lung metastasis models,respectively.Further mechanistic studies demonstrated that compound 55 mitigated TGF-β1-induced epithelial-mesenchymal transition(EMT).

Cell cycle regulation and anticancer drug discovery

Cellular growth,development,and differentiation are tightly controlled by a conserved biological mechanism:the cell cycle.This cycle is primarily regulated by cyclin-dependent kinase(CDK)-cyclin complexes,checkpoint kinases,and CDK inhibitors.Deregulation of the cell cycle is a hallmark of the transformation of normal cells into tumor cells.Given its importance in tumorigenesis,several cell cycle inhibitors have emerged as potential therapeutic drugs for the treatment of cancers-both as singleagent therapy and in combination with traditional cytotoxic or molecular targeting agents.In this review,we discuss the mechanisms underlying cell cycle regulation and present small-molecule anticancer drugs that are under development,including both pan-CDK inhibitors and CDK4/6-selective inhibitors.In addition,we provide an outline of some promising CDK inhibitors currently in preclinical and clinical trials that target cell cycle abnormalities in various cancers.

选择性Janus激酶1抑制药Upadacitinib和Filgotinib治疗类风湿性关节炎疗效和安全性的Meta分析

目的:系统评价两种选择性Janus激酶1(JAK-1)抑制药Upadacitinib和Filgotinib治疗类风湿性关节炎的疗效和安全性,为临床治疗提供循证参考。方法:计算机检索PubMed、Medline、Embase、Cochrane图书馆、中国生物医学文献数据库、中国期刊全文数据库、万方数据和中文科技期刊数据库,检索时限均为自建库起至2019年1月,收集在甲氨蝶呤或其他抗风湿类药物的基础上,安慰剂(对照组)对比Upadacitinib或Filgotinib(试验组)治疗类风湿性关节炎的随机对照试验(RCT),进行资料提取并采用Cochrane系统评价员手册5.1.0进行质量评价后,采用Rev Man 5.3统计软件对疗效[按美国风湿病协会标准判断病情缓解20%的患者比例(ACR20)、ACR50、ACR70、28个关节疾病活动度评分(DAS28)<3.2的患者比例]和安全性[不良事件(AE)发生率、严重不良事件(SAE)发生率、感染发生率、严重感染发生率、带状疱疹发生率、肝损害发生率]进行Meta分析。结果:共纳入8项RCT,合计2 738例患者。Meta分析结果显示,试验组患者ACR20[OR=3.37,95%CI(2.80,4.05),P<0.001]、ACR50[OR=3.78,95%CI(2.98,4.78),P<0.001]、ACR70[OR=4.31,95%CI(3.05,6.09),P<0.001]、DAS28<3.2分的患者比例[OR=3.86,95%CI(2.98,5.00),P<0.001]、AE发生率[OR=1.33,95%CI(1.11,1.61),P=0.002]和感染发生率[OR=1.43,95%CI(1.12,1.81),P=0.004]均显著高于对照组,其余指标比较差异均无统计学意义(P>0.05)。结论:JAK-1抑制药Upadacitinib和Filgotinib在提高类风湿性关节炎患者的ACR20、ACR50、ACR70、DAS28<3.2的患者比例等疗效指标方面较好;不会增加SAE、严重感染、带状疱疹与肝损害的发生率,但会增加患者AE与感染的风险。

分子对接与分子动力学模拟法探究PI3Kδ/度维利塞(Duvelisib)的选择性结合

大量研究表明磷脂酰肌醇3-激酶δ(PI3Kδ)与多种恶性肿瘤及免疫疾病的发生、发展密切相关,因此成为一个备受关注的药物靶点。伊德利塞(Idelalisib),PI3Kδ抑制剂,是首个被FDA批准上市的PI3K抑制剂,以此开启了PI3Kδ选择性抑制剂开发的热潮,但是严重的毒副作用阻碍了该类化合物的使用。随后,度维利塞(Duvelisib,IPI-145)于2018年被批准上市,度维利塞是PI3Kδ/γ选择性抑制剂,然而目前关于度维利塞的选择性PI3K抑制分子机制报道较少,且目前尚无度维利塞/PI3K复合物晶体结构报道。因此本文采用整合的计算机模拟策略来揭示度维利塞的选择性抑制机制:通过分子对接获得度维利塞与PI3K各亚型的合理结合构象;分子动力学模拟结合自由能计算揭示选择性产生的关键位点及热点氨基酸。目前,因与其他亚型之间高度的同源性与结构保守性,使得PI3Kδ选择性抑制剂开发受到极大挑战,本文以上市药物度维利塞为研究主体,有望为新型PI3Kδ选择性抑制剂的开发及合理药物设计提供一定的指导意义。

选择性CDK4/6抑制剂的研究进展

细胞周期蛋白依赖性激酶4/6(CDK4/6)与细胞周期蛋白D(cyclin D)结合后可形成cyclin D-CDK4/6复合物,该复合物通过调节细胞G1-S期转换可在细胞周期中起到关键的调控作用。选择性CDK4/6抑制剂能调控cyclin D-CDK4/6-INK4-Rb信号通路的异常活化,诱导肿瘤细胞出现G1细胞周期阻滞,从而可控制肿瘤细胞的增殖。近年来,选择性CDK4/6抑制剂已成为抗肿瘤药物研发的重要方向。本文对选择性CDK4/6抑制剂的作用机制及研究进展进行介绍。

靶向ATP活性位点的CDK5抑制剂研究进展

细胞周期蛋白依赖性激酶5(cyclin-dependentkinase5,CDK5)是一种丝氨酸/苏氨酸激酶,是CDKs家族的非典型成员,主要被非细胞周期蛋白p35或p39(及各自的截短产物p25或p29)激活,磷酸化下游底物,调控多种神经和非神经功能。研究表明CDK5的异常活化,与多种神经退行性疾病、癌症、糖尿病和炎症等疾病的发生发展密切相关。因此,靶向CDK5已成为多种疾病治疗的重要靶点。然而到目前为止,尚无选择性CDK5抑制剂处于临床阶段。而泛CDK抑制剂存在临床疗效不佳和因广泛抑制其他激酶而引起的不良反应,故临床试验进展缓慢。鉴于此,选择性CDK5抑制剂不论是对阐明CDK5的生物学功能,还是对探索CDK5抑制剂作为安全有效的临床治疗方案都具有重要意义。本文对CDK5的蛋白结构、生物功能、与疾病的关系进行简要概述,重点讨论了靶向ATP活性位点的CDK5抑制剂的结构类型和结合模式,并对开发选择性CDK5抑制剂的策略进行总结与展望。

口服选择性酪氨酸激酶2抑制剂氘可来昔替尼

氘可来昔替尼(deucravacitinib)是第1个获批上市的新型口服选择性酪氨酸激酶2(tyrosine kinase 2,TYK2)抑制剂,用于治疗多种自身免疫疾病,临床应用前景广泛.本文对deucravacitinib的作用机制、研发历程、临床评价、同靶点药物研究进展等进行综述.

Discovery of a series of dimethoxybenzene FGFR inhibitors with 5H-pyrrolo[2,3-b]pyrazine scaffold: structure–activity relationship, crystal structural characterization and in vivo study

Genomic alterations are commonly found in the signaling pathways of fibroblast growth factor receptors(FGFRs). Although there is no selective FGFR inhibitors in market, several promising inhibitors have been investigated in clinical trials, and showed encouraging efficacies in patients. By designing a hybrid between the FGFR-selectivity-enhancing motif dimethoxybenzene group and our previously identified novel scaffold, we discovered a new series of potent FGFR inhibitors, with the best one showing sub-nanomolar enzymatic activity. After several round of optimization and with the solved crystal structure, detailed structure–activity relationship was elaborated. Together with in vitro metabolic stability tests and in vivo pharmacokinetic profiling, a representative compound(35) was selected and tested in xenograft mouse model, and the result demonstrated that inhibitor 35 was effective against tumors with FGFR genetic alterations, exhibiting potential for further development.