CN109999041A - 吩噻嗪衍生物对malt1蛋白酶的选择性抑制 - Google Patents
吩噻嗪衍生物对malt1蛋白酶的选择性抑制 Download PDFInfo
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- CN109999041A CN109999041A CN201811390121.6A CN201811390121A CN109999041A CN 109999041 A CN109999041 A CN 109999041A CN 201811390121 A CN201811390121 A CN 201811390121A CN 109999041 A CN109999041 A CN 109999041A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
- A61K31/5415—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with carbocyclic ring systems, e.g. phenothiazine, chlorpromazine, piroxicam
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
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Abstract
本发明涉及用于治疗癌症的化合物或所述化合物的可药用盐、前药、对映体、非对映体、外消旋混合物、结晶形式、无定形形式、非溶剂化形式或溶剂合物,其中所述癌症依赖于MALT1蛋白酶的蛋白水解活性,并且其中所述化合物具有通式(I),在通式(I)中,X是N或C;Y是S、O、SO2、SO、NH、CO、CH2、CH=CH或CH2‑CH2;()z是C1‑C5直链或支链烷基链;A是NR3R4、OR5或HET;R1和R2在每次出现时独立地选自‑H、‑CH3、‑OH、‑OCH3、‑SCH3、‑F、‑Cl、‑CF3、‑NH2以及‑COOH;R3、R4和R5是H或C1‑C5直链或支链烷基,并且HET是5、6或7元杂环,其中环原子可以是C、O、N或S,所述环可以是饱和的或芳族的,并且所述环可以被H或C1‑C5直链或支链烷基取代。本发明化合物还可用于治疗MALT1依赖性免疫疾病。
Description
本申请为2012年8月1日提交的申请号为201280036975.9的名称为“吩噻嗪衍生物对MALT1蛋白酶的选择性抑制”的申请的分案申请。
技术领域
本发明涉及用于治疗癌症的化合物或所述化合物的可药用盐、前药、对映体、非对映体、外消旋混合物、结晶形式、无定形形式、非溶剂化形式(unsolvated form)或溶剂合物,其中所述癌症依赖于MALT1蛋白酶的蛋白水解活性,并且其中所述化合物具有通式(I):
其中,X是N或C;Y是S、O、SO2、SO、NH、CO、CH2、CH=CH或CH2-CH2;()z是C1-C5直链或支链烷基链;A是NR3R4、OR5或HET;R1和R2在每次出现时独立地选自-H、-CH3、-OH、-OCH3、-SCH3、-F、-Cl、-CF3、-NH2以及-COOH;R3、R4和R5是H或C1-C5直链或支链烷基,并且HET是5、6或7元杂环,其中环原子可以是C、O、N或S,所述环可以是饱和的或芳族的,并且所述环可以被H或C1-C5直链或支链烷基取代。本发明化合物还可用于治疗MALT1依赖性免疫疾病。
背景技术
在本说明书中,引用了许多文件,包括专利申请和制造商的说明书。虽然认为这些文件的公开内容不与本发明的专利性相关,但是通过引用以所述公开内容的整体并入本文。更具体地,全部所引用文件通过引用并入的程度如同各个文件特别地且单独地指明通过引用所并入的程度。
粘膜相关淋巴组织淋巴瘤转运蛋白1(mucosa-associated lymphoid tissuelymphoma translocation protein 1,MALT1)是由T细胞受体刺激而活化的功能性半胱氨酸蛋白酶。MALT1在arg439之后迅速切割A20(TNFAIP),这减弱其NF-κB抑制剂功能(Coornaert等.(2008),Nature Immun.9:263-271)。
在抗原刺激后,MALT1是控制淋巴细胞活化、存活及分化的上游NF-κB信号传导的关键介质1。MALT1与CARMA1(也称为CARD11)和BCL10一起组装成所谓的CBM复合物,该复合物桥连邻近的抗原受体信号传导事件与IκB激酶(IKK)复合物,IKK复合物是典型NF-κB途径的看门复合物(gatekeeper)2。在T细胞抗原受体(TCR)/CD28共刺激后,MALT1充当将其他关键信号传导分子(例如TRAF6、CASP8和A20)募集至CBM复合物的蛋白质支架(proteinscaffold)1。此外,由E3连接酶TRAF6催化的MALT1中的共价泛素修饰促进两种下游蛋白激酶复合物TAB2-TAK1和NEMO-IKKα/β的缔合(association),这最终导致IKK活化3。
MALT1含有表现出与来自哺乳动物的胱天蛋白酶(caspase)和来自植物和真菌的间胱天蛋白酶(metacaspase)高同源性的对胱天蛋白酶(paracaspase)结构域4。正如间胱天蛋白酶,MALT1在精氨酸残基之后切割底物,表明酶促切割活性与胱天蛋白酶的很大区别在于一般需要P1位的天冬氨酸5。MALT1的蛋白水解活性在TCR/CD28刺激之后诱导,这促进底物BCL10、A20和CYLD的切割6-8。由拮抗性四肽Z-VRPR-FMK(其最初设计为植物间胱天蛋白酶的抑制剂)对MALT1蛋白酶活性的抑制减弱T细胞中的最佳NF-κB活化和IL-2产生7,9。类似地,催化半胱氨酸464的突变使MALT1蛋白水解地失活并且还在补给MALT1缺陷型T细胞之后减弱IL-2产生9。
MALT1蛋白酶活性的失调在多种疾病的产生中起着重要作用,特别是依赖于MALT1蛋白酶之蛋白水解活性的癌症和MALT1依赖性免疫疾病。MALT1的肿瘤促进作用已见于弥漫性大B细胞淋巴瘤(DLBCL)和粘膜相关淋巴组织(MALT)淋巴瘤亚组中10。通过基因表达分析,可以将DLBCL分为不同的实体,并且含量最高的亚型是“活化的B细胞样”(ABC-)DLBCL和“生发中心B细胞样”(GCB-)DLBCL11-15。基于基因表达特征,ABC-DLBCL亚型来自通过其B细胞抗原受体(BCR)刺激的B淋巴细胞。鉴于约30%的5年存活率,ABC-DLBCL患者具有最差的预后,这反映出ABC-DLBCL细胞的侵袭性临床行为16。ABC-DLBCL细胞的特点(hallmark)是NF-κB信号传导通路的组成性活化,而GCB-DLBCL细胞并非如此11,17。明显的分子畸变的鉴定表明,ABC-DLBCL中的促生存NF-κB信号传导是由BCR信号传导的失调导致的。虽然一些ABC-DLBCL患者携带致瘤的CARMA1突变18,但是大多数ABC-DLBCL细胞的特征在于,慢性的活性BCR信号传导和突变经常见于BCR近端调节因子CD79A和B19。与对BCR信号传导的需求相符,RNA干扰筛选将CARMA1、BCL10或MALT1鉴定为ABC-DLBCL的NF-κB活化、存活和生长的关键调节因子10。此外,用Z-VRPR-FMK抑制MALT1蛋白水解活性特别地抑制ABC-DLBCL细胞中的NF-κB依赖性基因表达并发挥毒性作用20,21。Ferch等,(2009),J.Exp.Med.206:2313-2320示出侵袭性的活化的B细胞样(ABC)弥散性大B细胞淋巴瘤(DLBCL)细胞(而非生发中心B细胞样(GCB)DLBCL)组成性地具有组装的CARD11-BCL10-MALT1(CBM)复合物,该复合物连续地并且选择性地加工A20。MALT1的抑制阻断A20和BCL10切割,降低NFκB活性,并且降低靶向BCLXL(BCL2L1)、IL6和IL10的NF-κB的表达。抑制MALT1对胱天蛋白酶导致ABC-DLBCL细胞死亡和生长停滞。Ferch等(2009)总结,MALT1对胱天蛋白酶活性具有促生长作用,特别是在ABC-DLBCL细胞中,并且提出MALT1蛋白酶活性是ABC-DLBCL之药物治疗的潜在靶标。
MALT淋巴瘤是B细胞淋巴细胞的癌症。其通常影响60多岁的年长者。大多数非霍奇金淋巴瘤(Non-Hodgkin Lymphoma,NHL)从淋巴结开始,但MALT淋巴瘤从称为粘膜相关淋巴组织(MALT)的淋巴组织开始。胃是最常见的发生MALT淋巴瘤的区域,但MALT淋巴瘤也可能从其他器官(例如肺、甲状腺、唾液腺或肠)开始。因为MALT淋巴瘤在淋巴结外发生,所以也将其称为淋巴结外淋巴瘤。由于幽门螺杆菌(Helicobacter pylori)的存在,胃MALT淋巴瘤常常与慢性炎症相关(72%至98%)(Parsonnet J(1994).N Engl J Med 330(18):1267-71)。通过用食道、胃、十二指肠镜检查(EGD,上部内窥镜检查法)对怀疑病变进行生物活检来作出初始诊断。同时还对幽门螺杆菌进行测试以检测该微生物的存在。在另一些部位,慢性免疫刺激也被怀疑为发病机理(例如,慢性自身免疫病(例如,肖格伦综合征(syndrome)和桥本甲状腺炎(Hashimoto′s thyroiditis))与唾液腺和甲状腺的MALT淋巴瘤之间的关系)。在MALT淋巴瘤中,频繁的易位(translocation)t(11;18)(q21;q21)在MALT1的C末端(包括对胱天蛋白酶结构域)与IAP2的N末端之间产生融合22。IAP2-MALT1融合蛋白的对胱天蛋白酶结构域催化NIK的切割,从而增强非典型NF-κB活化,其赋予抗凋亡性23。
将对抗MALT1对胱天蛋白酶的新试剂组合在一起可以有益于治疗与失调的MALT1活性相关的淋巴瘤和MALT1依赖性免疫疾病。特别地,ABC-DLBCL患者在总计5年的存活率仅有约30%,这强调了对替代治疗选择(特别是对于该淋巴瘤类型)的清楚的需要16。因此,本发明的一个目的是提供对抗MALT1的新试剂,其可用于治疗上述疾病。
发明内容
因此,本发明在第一实施方案中涉及用于治疗癌症的化合物或所述化合物的可药用盐、前药、对映体、非对映体、外消旋混合物、结晶形式、无定形形式、非溶剂化形式或溶剂合物,其中所述癌症依赖于MALT1蛋白酶的蛋白水解活性,并且其中所述化合物具有通式(I):
其中,X是N或C;Y是S、O、SO2、SO、NH、CO、CH2、CH=CH或CH2-CH2;()z是C1-C5直链或支链烷基链;A是NR3R4、OR5或HET;R1和R2在每次出现时独立地选自-H、-CH3、-OH、-OCH3、-SCH3、-F、-Cl、-CF3、-NH2以及-COOH;R3、R4和R5是H或C1-C5直链或支链烷基,并且HET是5、6或7元杂环,其中环原子可以是C、O、N或S,所述环可以是饱和的或芳族的,并且所述环可以被H或C1-C5直链或支链烷基取代。
本文中使用的术语“依赖于MALT1蛋白酶之蛋白水解活性的癌症”定义了部分地或主要地由MALT1的非生理性升高的(蛋白水解)活性导致的癌症。MALT-1的酶促活性包含半胱氨酸蛋白酶活性(EC 3.4.22.-半胱氨酸内肽酶)。如因所附实施例而明显的,本发明人发现本发明化合物特异性地抑制MALT1的活性。如在上文中详细讨论的,MALT1的活性是抗原受体刺激的T细胞中最佳NF-κB活化和IL-2产生的原因。这表明,MALT1活性对于生理学上的淋巴细胞活化而言是必需的。因此,依赖于MALT1蛋白酶之蛋白水解活性的癌症优选地为依赖于MALT1蛋白酶之蛋白水解活性的淋巴瘤。依赖于MALT1蛋白酶之蛋白水解活性的淋巴瘤的优选实例是在下文中更详细地讨论的MALT淋巴瘤和弥散性大B细胞淋巴瘤的活化B细胞亚型(ABC亚型)。
本发明还包括通式(I)化合物的可药用盐、前药、对映体、非对映体、外消旋混合物、结晶形式、非结晶形式、无定形形式、非溶剂化形式和溶剂合物。
本文使用的术语“可药用盐”包括通式(I)化合物的盐,其使用相对无毒(即,可药用)的酸或碱来制备,这取决于存在于本发明化合物上的特定取代基。例如,如果本发明化合物含有酸性官能团,则可以通过使这样的化合物的中性形式与足量的期望的碱(纯的或在合适的惰性溶剂中)接触来获得碱加成盐。可药用碱加成盐的非限制性实例包括钠盐、钾盐、钙盐、铵盐、有机胺盐或镁盐,或者类似的盐。如果本发明化合物含有碱性官能团,则可以通过使这样的化合物的中性形式与足量的期望的酸(纯的或在合适的惰性溶剂中)接触来获得酸加成盐。可药用酸加成盐的非限制性实例包括衍生自例如以下之无机酸的那些:盐酸、氢溴酸、硝酸、碳酸、磷酸、部分中和的磷酸、硫酸、部分中和的硫酸、氢碘酸或亚磷酸等,以及衍生自例如以下之相对无毒的有机酸的盐:乙酸、丙酸、异丁酸、马来酸、丙二酸、苯甲酸、琥珀酸、辛二酸、富马酸、扁桃酸、邻苯二甲酸、苯磺酸、对甲苯磺酸、柠檬酸、酒石酸、甲磺酸等。还包括氨基酸(例如精氨酸等)的盐和有机酸(例如葡萄糖醛酸或半乳糖醛酸等)的盐。本发明的某些特定化合物可包含碱性官能团和酸性官能团二者,其允许化合物转化为碱加成盐或酸加成盐。本发明化合物的中性形式可以通过以下来再生:使所述盐与碱或酸相接触,并以常规方式分离母体化合物。出于本发明的目的,所述化合物的母体形式在某些物理性质方面(例如在极性溶剂中的溶解度)不同于多种盐形式,然而在其他方面,所述盐等同于所述化合物的母体形式。
本发明化合物可以具有手性的或不对称的碳原子(光学中心)和/或双键。本发明包括外消旋物、非对映体、几何异构体以及单个光学异构体。本发明化合物可以以非溶剂化形式以及溶剂化形式存在,包括水合形式。一般来说,溶剂化形式等同于非溶剂化形式并且由本发明所涵盖。本发明化合物还可以以多种结晶形式或无定形形式存在。
除盐形式以外,本发明化合物可以为前药形式。本发明化合物的前药是在生理条件下易于经历化学变化以提供本发明化合物的那些化合物。此外,可以在离体环境下通过化学方法或生物化学方法将前药转化为本发明化合物。例如,当放置在具有合适的酶或化学试剂的透皮贴片储库(reservoir)中时,前药可以缓慢地转化为本发明化合物。
本文所述的本发明化合物可以以合适的剂量施用于对象。本发明化合物优选地施用于哺乳动物,例如家畜和宠物。家畜和宠物的非限制性实例是猪、牛、水牛(buffalo)、绵羊、山羊、兔、马、驴、鸡、鸭、猫、狗、纯种猪(genuine pig)或仓鼠。最优选将其施用于人。优选的施用方式取决于本发明化合物(具有通式(I))的形式。如上文所述,具有通式(I)的化合物可以为以下形式:可药用盐、前药、对映体、非对映体、外消旋混合物、结晶形式、非结晶形式、无定形形式、非溶剂化形式或溶剂合物。本发明化合物可以以剂量单位制剂(任选地还包含常规可药用赋形剂)经口、肠胃外例如皮下、静脉内、肌内、腹膜内、鞘内、经皮、经粘膜、硬膜下、通过离子电渗疗法(iontopheresis)局部或外用、舌下、通过吸入喷雾剂、气雾剂或直肠等来施用。
可以使用一种或更多种生理学载体或赋形剂将用于根据本发明使用的本发明化合物配制为药物组合物,参见例如,Ansel等,“Pharmaceutical Dosage Forms and DrugDelivery Systems”,第7版,Lippincott Williams & Wilkins Publishers,1999。
就经口施用而言,本发明的药物组合物可以采用例如片剂或胶囊剂的形式,其通过常规方法用例如以下可药用赋形剂制备:粘合剂(例如,预凝胶化的玉米淀粉、聚乙烯吡咯烷酮、羟丙基甲基纤维素)、填充剂(例如,乳糖、微晶纤维素、磷酸氢钙)、润滑剂(例如,硬脂酸镁、滑石粉、二氧化硅)、崩解剂(例如,马铃薯淀粉、羟基乙酸淀粉钠)或润湿剂(例如,月桂基硫酸钠)。所述药物组合物可以与生理学上可接受的载体一起施用于患者。在一个特定实施方案中,术语“可药用”意指经管理部门或其他一般性公知的药典批准用于动物,更特别地用于人。术语“载体”是指与治疗剂一起施用的稀释剂、辅料、赋形剂或载剂。这样的可药用载体可以是无菌液体,例如水和油,包括石油、动物油、植物油或合成来源的油(例如花生油、豆油、矿物油、芝麻油等)。当经静脉内施用药物组合物时,水是优选的载体。盐溶液和水性葡萄糖及甘油溶液也可用作液体载体,特别是对于可注射溶液而言。合适的药物赋形剂包括淀粉、葡萄糖、乳糖、蔗糖、明胶、麦芽、稻米、面粉、白垩、硅胶、硬脂酸钠、单硬脂酸甘油酯、滑石粉、钠离子、脱脂乳粉、甘油、丙烯、乙二醇、水、乙醇等。如果需要的话,组合物还可以包含少量的润湿剂或乳化剂,或者pH缓冲剂。这些组合物的形式可以为软膏剂、溶液剂、混悬液、乳剂、片剂、丸剂、胶囊剂、散剂、持续释放制剂等。优选的形式是软膏剂。所述组合物可以与传统的粘合剂和载体(例如甘油三酯)一起配制成栓剂。经口制剂可以包括标准载体,例如药用级别的甘露醇、乳糖、淀粉、硬脂酸镁、糖精钠、纤维素、碳酸镁等。E.W.Martin的“Remington′s Pharmaceutical Sciences”中描述了合适的药物载体的实例。这样的组合物将包含治疗有效量的前述化合物(优选为纯化形式)和合适量的载体以提供用于适当施用于患者的形式。该制剂应适于施用形式。
用于经口施用的液体制剂可以为例如溶液剂、糖浆剂或混悬剂形式,或者可以呈现为干燥产物用于在使用之前与水或其他合适的载剂组合。这样的液体制剂可以通过常规方法用例如以下的可药用添加剂来制备:助悬剂(例如,山梨醇、糖浆、纤维素衍生物、氢化可食用脂肪)、乳化剂(例如,卵磷脂、阿拉伯胶)、非水性载剂(例如,杏仁油、油性酯、乙醇、经分馏的植物油)、防腐剂(例如,对羟基碳酸甲酯或丙酯、山梨酸(soric acid))。在认为适当时,该制剂还可以包含缓冲盐、调味剂、着色剂以及甜味剂。用于经口施用的制剂可以合适地配制以提供本发明药物组合物的受控释放。
就通过吸入施用而言,本发明的药物组合物方便地从加压包装(pressurisedpack)或雾化器以气雾剂喷剂的呈现形式递送,同时使用合适的抛射剂(例如,二氯二氟甲烷、三氯氟甲烷、二氯四氟乙烷、二氧化碳或其他合适的气体)。在加压气雾剂的情况下,可以通过提供阀以递送计量的量来确定剂量单位。用于吸入器或吹入器中的例如明胶胶囊和盒(cartridge)可以配制以包含本发明的药物组合物与合适的粉末基质(例如乳糖或淀粉)的粉末混合物。
本发明的药物组合物可以配制用于通过注射(例如,通过大量注射或连续输注)进行肠胃外施用。注射部位包括静脉内、腹膜内或皮下。用于注射的制剂可以与所添加的防腐剂以单位剂型存在(例如,在药瓶(phial)中、在多剂量容器中)。本发明的药物组合物可以采用这样的形式:混悬剂、溶液剂或者油性或水性载剂中的乳剂,并且可以包含配制用试剂(formulatory agent),例如助悬剂、稳定剂或分散剂。或者,该试剂可以为粉末形式用于在使用之前与合适的载剂(例如,灭菌的无热原水)组合。通常来说,用于静脉内施用的组合物是在无菌等渗水性缓冲剂中的溶液。在必要时,所述组合物还可以包含增溶剂和局部麻醉剂(例如利诺卡因)以缓解注射部位的疼痛。一般来说,成分单独提供或者混合在单位剂型中一起提供,例如,作为在密封容器(例如安瓿或小袋,其示出活性剂的量)中的冻干粉末或无水浓缩物。在通过输注施用所述组合物时,其可以使用含有灭菌药用级别的水或盐水的输注瓶来配制。在通过注射施用所述组合物时,可以提供用于注射的灭菌水或盐水的安瓿使得可以在施用之前混合这些成分。
如果需要的话,本发明的药物组合物还可以存在于包装或分配器装置中,所述包装或分配器装置可以含有一种或更多种包含所述试剂的单位剂型。所述包装可以例如包含金属或塑性薄片,例如罩板包装(blister pack)。所述包装或分配器装置可以附有施用说明书。
本发明的药物组合物可以作为单独的活性剂施用或者可以与其他试剂组合施用。
根据第一实施方案,优选X是N。此外,优选Y是S。()z优选地为直链的C1-C5烷基链,更优选地为直链的C1-C3烷基链。R1优选为-H;并且R2优选为-H或-SCH3。优选地,优选的实施方案可以彼此独立存在。在另一个优选的实施方案中,存在所有优选实施方案的特征。
因此,根据一个优选的实施方案,用于根据本发明所述用途的化合物具有上述式(I),其中在式(I)中,X是N;Y是S;()z是直链的C1-C5烷基链。R1是-H;并且R2是-H或-SCH3。
根据本发明的一个更优选的实施方案,用于根据本发明所述用途的化合物具有上述式(I),其中在式(I)中,A是HET,并且HET是5元至7元碳环,其任选地插入有NR3。
鉴于此,优选HET是6元碳环。甚至更优选HET是插入有NR3的6元碳环,其中R3是CH3。
根据本发明的另一个更优选的实施方案,用于根据本发明所述用途的化合物具有上述式(I),其中在式(I)中,A是NR3R4,R3是H或CH3,并且R4是-CH3。
鉴于此,最优选R3和R4是-CH3。
根据本发明的另一个更优选的实施方案,用于根据本发明所述用途的化合物具有上述式(I),其中在式(I)中,A是NR3R4,其中R3是CH3,R4是-CH3、-C2H5或C3-C5的直链烷基链,该链可以插入有O、N或S,并且其与()z的碳原子形成饱和环。
鉴于此,最优选R4是-CH3。
根据本发明的一个甚至更优选的实施方案,用于根据本发明所述用途的化合物具有上述式(I),其中饱和环是插入有N的5元至7元碳环。
鉴于此,优选任选地插入有N的5元至7元亚烷基环是6元亚烷基环。还优选所述饱和环是5元至7元饱和碳环(未插入有N),更优选为6元饱和碳环(未插入有N)。
根据本发明的一个更优选的实施方案,用于根据本发明所述用途的化合物具有上述式(I),其中在式(I)中,A是HET,并且HET是N-甲基哌啶-3-基。
根据本发明的另一个更优选的实施方案,用于根据本发明所述用途的化合物具有上述式(I),其中在式(I)中,(a)Z=3,A是NR3R4,并且R3和R4是-CH3;(b)Z=1,A是N-甲基哌啶-3-基;或(c)Z=2,A是N-甲基哌啶-2-基。
根据本发明的最优选的实施方案,所述用于根据本发明所述用途的化合物是:
在本领域中将式(II)化合物称为甲哌啶嗪(mepazine)。甲哌啶嗪是吩噻嗪,其最初用作镇静剂(Lord and Archibald(1957),Can J Comp Med Vet Sci.,21(11):391-394)。
在本领域中将式(III)化合物称为甲硫哒嗪(thioridazine)。甲硫哒嗪也属于吩噻嗪药物组。在本领域中已知甲硫哒嗪是抗精神病药(antipsychotic drug)并且广泛用于治疗精神分裂症和精神病。
在本领域中将式(IV)化合物称为普马嗪(promazine)。普马嗪是吩噻嗪的衍生物。普马嗪在本领域中用作抗精神病药,例如用于治疗精神分裂症。
在所附实施例中分析的三种吩噻嗪衍生物(PD)全部都已进行临床试验并且用作抗精神病药和/或镇静药,并且认为该活性主要基于其作为多巴胺D2受体拮抗剂之作用的能力30。在50年代末和60年代初,已经以商标名Pacatal对甲哌啶嗪作为抗精神病药和镇静药物进行了评价。但是一些临床研究证实了抗精神病作用,而另一些未能证实25,31。报道了一些副作用,包括甲哌啶嗪治疗后哮喘发作减少,表明了某些免疫抑制活性31。就本发明人所知,没有关于对癌症患者之潜在有益作用的观察结果的报道。研究设计和群组大小(cohort size)都不允许得出甲哌啶嗪、甲硫哒嗪和普马嗪可以特异性地抑制MALT1的任何结论。甲硫哒嗪(商标名Mellaril)仍然在市售,但处方保留用于治疗对其他抗精神病药无响应的精神分裂症患者。还认为甲硫哒嗪有益于其他医学应用,因其对不同的癌细胞系发挥毒性作用29,32。但是,本发明人未知任何示出或表明甲硫哒嗪依赖于蛋白水解MALT1活性而对癌细胞系发挥毒性作用的现有技术。此外,认为甲硫哒嗪是用于治疗肺结核或疟疾的候选药物,但目前未知其抗微生物和抗寄生虫作用的原因33,34。对MALT1依赖性ABC-DLBCL示出最弱毒性的普马嗪(商标名Sparine)仍用于治疗不安行为(restless behavior)。
因此,式(II)、(III)和(IV)的化合物最初在本领域中都用作抗精神病药。在所附实施例中,将甲哌啶嗪、甲硫哒嗪和普马嗪鉴定为MALT1的三种小分子抑制剂。就本发明人所知,这些化合物都并非已知抑制MALT1蛋白酶的活性。在本发明的实施例中举例说明的结果首次示出式(II)、(III)和(IV)的化合物可用于治疗依赖于MALT1蛋白酶之蛋白水解活性的癌症。
根据本发明的一个优选的实施方案,依赖于MALT1蛋白酶之蛋白水解活性的癌症是MALT淋巴瘤或弥散性大B细胞瘤的活化B细胞亚型。
如在上文中所述的,弥散性大B细胞淋巴瘤(DLBCL)为侵袭性淋巴瘤类型。基于DLBCL遗传活性鉴定的DLBCL的一个主要亚型是弥散性大B细胞淋巴瘤的B细胞亚型(ABC-DLBCL)。如在上文中描述的,Ferch等(2009),J.Exp.Med.2006:2313-2320示出,侵袭性活化B细胞样(ABC)弥散性大B细胞淋巴瘤(DLBCL)细胞具有组成性装配的CARD11-BCL10-MALT1(CBM)复合物,其连续地并且选择性地加工A20。此外,抑制MALT1对胱天蛋白酶导致ABC-DLBCL细胞死亡和生长停滞。因此,在下文中示出吩噻嗪衍生物甲哌啶嗪、甲硫哒嗪和普马嗪特异性地抑制MALT1的实施例首次表明可以通过使用本发明化合物来治疗ABC-DLBL。
如在上文中描述的,MALT淋巴瘤是B细胞淋巴细胞的癌症。大多数NHL始于淋巴结,但MALT淋巴瘤始于粘膜相关淋巴组织(MALT)。MALT淋巴瘤通常始于机体的已被感染的区域或者当人具有影响该区域的自身免疫病症时的区域。影响胃的MALT淋巴瘤的大多数情况与被称为幽门螺杆菌的细菌感染相关。在另一些部位,慢性免疫刺激也被怀疑为发病机理(例如,慢性自身免疫病(例如,肖格伦综合征和桥本甲状腺炎)与唾液腺和甲状腺的MALT淋巴瘤之间的关系)。已鉴定出了与MALT淋巴瘤相关的三种易位:即,t(11;18)(q21;q21),其使得产生API2-MLT融合基因;t(1;14)(p22;q32),其使BCL10失调;以及t(14;18)(q32;q21),其使MALT1失调。认为三种易位全都启动相同的途径,即,API2-MALT途径。因此,下文中示出吩噻嗪衍生物甲哌啶嗪、甲硫哒嗪和普马嗪特异性地抑制MALT1的实施例首次表明可以通过使用本发明化合物来治疗MALT淋巴瘤。
根据本发明,已经将吩噻嗪衍生物(PD)鉴定为第一类小分子抑制剂,其有效地并且选择性地抑制重组的和细胞的MALT1蛋白酶的蛋白水解活性。如可以从实施例中得知的,用甲哌啶嗪、甲硫哒嗪和普马嗪获得了最佳抑制活性。三种PD全部示出分别干扰活化T细胞或ABC-DLBCL细胞的诱导性或组成性MALT1活性。此外,这些PD导致受损的T细胞活化以及DLBCL细胞之ABC亚型的选择性降低的活力,已示出主要依赖于MALT1活性的过程9,20,21。因此,细胞数据还证实了PD作为药理学MALT1抑制剂的有效性。
最初测试了不同的测定条件和广谱蛋白酶抑制剂的作用以更详细地表征重组全长MALT1的切割活性。有趣的是,MALT1的蛋白水解活性类似于拟南芥(Arabidopsisthaliana)间胱天蛋白酶AtMC4和95,强调对胱天蛋白酶结构域与间胱天蛋白酶结构域之间的结构同源性导致类似的底物结合性质和切割性质。由于MALT1是与其他人胱天蛋白酶相比唯一具有非常特殊之性质的人对胱天蛋白酶,所以根据本发明定义的特异性抑制剂明显是用于其致癌活性之选择性失活的有前景候选。选择性是关键的,这是因为通过胱天蛋白酶而非MALT1的抑制减弱凋亡的进行将可能引起淋巴瘤治疗无法耐受的不良作用。事实上,所测试的全部PD都表现出对MALT1的高偏好性,并且不对启动子胱天蛋白酶CASP8和执行子胱天蛋白酶CASP3起作用。此外,由于CASP8与MALT1相缔合,并且这是T细胞的NF-κB信号传导所需要的27,所以由PD引起的CASP8抑制的明显缺乏还强调对蛋白水解MALT1活性的需要以引起最佳T细胞活化。即使在相对短的PD孵育之后细胞MALT1活性的强抑制也清楚地表明这些物质直接影响MALT1蛋白酶。
此外,本发明的MALT1抑制化合物对T细胞活化的抑制作用表明例如在变态反应和哮喘的治疗中作为温和免疫抑制剂的潜在医疗用途。
因此,本发明还包括本发明的化合物,其用于治疗MALT1依赖性免疫疾病。
根据本发明的一个优选的实施方案,MALT1依赖性免疫疾病是变应性炎症。
本文还描述了在对象中依赖于MALT1蛋白酶之蛋白水解活性来治疗癌症的方法,该方法包括向对象施用药用有效量的本发明化合物。鉴于此,所述依赖于MALT1蛋白酶之蛋白水解活性的癌症优选为MALT淋巴瘤或弥散性大B细胞淋巴瘤的活化B细胞亚型。此外,所述对象优选为哺乳动物,并且更优选为人。
此外,本文中描述了在对象中治疗MALT1依赖性免疫疾病的方法,该方法包括向对象施用药用有效量的本发明化合物。鉴于此,MALT1依赖性免疫疾病优选为变应性炎症。所述MALT1依赖性免疫疾病还可以是T细胞驱动的疾病,其中例如在实施例5中,T细胞应答被化合物所抵消。鉴于此,MALT1依赖性免疫疾病可以是免疫系统的超敏感性或慢性炎症,例如变态反应(如所提到的)或哮喘。此外,MALT1依赖性免疫疾病可以是自身免疫病,其包括但不限于以下疾病:例如,多发性硬化(multiple sclerosis)、炎性肠病(inflammatorybowel disease)(例如,克罗恩病(Crohn’s disease)、溃疡性结肠炎(ulcerativecolitis))、红斑狼疮(lupus erythematosus)、银屑病(psoriasis)、慢性阻塞性肺病(chronic obstructive pulmonary disease)、类风湿性关节炎(rheumatoid arthritis)或银屑病关节炎(psoriatic arthritis)。此外,所述对象优选为哺乳动物,并且更优选为人。
本文中的上述优选实施方案还适用于本文所述的治疗方法。
附图说明
图1:用于高通量筛选(HTS)的体外MALT1切割测定的建立。(A)MALT1蛋白酶测定的方案。由针对含有BCL10衍生之MALT1切割位点的荧光肽Ac-LRSR-AMC的GSTMALT1之蛋白水解作用引起的释放荧光团AMC导致荧光增加。(B)MALT1切割反应的动力学。在30℃,将从细菌表达的纯化重组GSTMALT1与50μM的Ac-LRSR-AMC孵育1小时,并通过测量AMC荧光的增加来确定蛋白水解活性。而催化失活的MALT1 C453A未能切割底物,用1nM的抑制肽Z-VRPR-FMK来抑制导致MALT1活性降低约50%。(C)MALT1被Z-VRPR-FMK抑制。增加量的肽导致MALT1活性的总体损失。就数据评价而言,将未经处理之对照的相对荧光设置为100%,从而计算经抑制剂处理之孔的值。(D)泛胱天蛋白酶抑制剂(pan-caspase inhibitor)Ac-DEVD-CHO未显著地作用于MALT1,即使在200μM时也是如此。(E)使用不同蛋白酶抑制剂进行的MALT1对胱天蛋白酶的酶促表征。MALT1活性因常见浓度的半胱氨酸蛋白酶抑制剂抗蛋白酶(antipain)(1μM)和胰凝乳蛋白酶抑制剂(chymostatin)(100μM)而降低,而未因高浓度的E-64(100μM)或低浓度的亮肽素(leupetin)(1μM)而降低。天门冬氨酰蛋白酶抑制剂胃蛋白酶抑制剂A(Pepstatin A)(100μM)、丝氨酸蛋白酶抑制剂抑肽酶(5μg/ml)和丝氨酸/半胱氨酸蛋白酶抑制剂TLCK(1μM)对MALT1活性没有影响。将抑制谱与拟南芥间胱天蛋白酶AtMC4和AtMC9进行比较(参见图9)。图示出至少三个独立实验的平均值,误差棒表示SD。
图2:通过HTS鉴定的吩噻嗪衍生物抑制MALT1活性。(A)鉴定为潜在MALT1抑制剂的PD的化学结构。化合物A(甲哌啶嗪;10-[(1-甲基-3-哌啶基)甲基]-10H-吩噻嗪乙酸酯)、B(2-氯吩噻嗪)和C([2-(3-异丁氧基-10H-吩噻嗪-10-基)乙基]二甲胺)是吩噻嗪衍生物(PD),化合物D是结构性PD相关物(structual PD relative)。(B)虽然用5μM至50μM的增加量的PD的治疗导致GSTMALT1活性的剂量依赖性降低,但酶促CASP8作用并未显著降低。(C)用1μM、5μM和20μM吩噻嗪以剂量依赖的方式抑制GSTMALT1活性。图(在B中)示出两个独立实验中的一个代表性实验或者至少3个独立实验(在C中)的平均值,误差棒表示SD。
图3:甲哌啶嗪、甲硫哒嗪和普马嗪的选择性MALT1抑制。(A)三种抑制化合物的分子结构。这三种化合物都在氮处具有短疏水侧链,该侧链具有类似原子组成和空间。(B)甲哌啶嗪、甲硫哒嗪和普马嗪的剂量响应曲线和IC50值。(C)甲哌啶嗪充当非竞争性MALT1抑制剂。在存在或不存在1μM甲哌啶嗪的条件下,用增加浓度的LRSR-AMC底物来测定米氏动力学(Michaelis-Menten kinetic)。甲哌啶嗪降低MALT1的VMAx而非KM。(D)甲哌啶嗪充当可逆的MALT1抑制剂。用甲哌啶嗪(10μM、20μM或50μM)将与谷胱甘肽琼脂糖珠相偶联的GSTMALT1处理30分钟。在切割反应开始之前,将所述珠清洗0、3或6次之后测定MALT1活性。(E)PD是选择性的MALT1抑制剂,并且在高达50μM的浓度下未能显著地抑制CASP3和8的活性。数据示出至少3个独立实验的平均值,并且误差棒表示SD。
图4:在原代小鼠CD4+T细胞、人PBMC和Jurkat T细胞中,甲哌啶嗪和甲硫哒嗪介导的MALT1抑制导致T细胞活化减弱。(A)Jurkat T细胞不予处理或者与10μM的甲哌啶嗪或甲硫哒嗪孵育3小时,然后不予刺激或者用抗CD3/CD28刺激15分钟、30分钟、60分钟、90分钟和120分钟。甲哌啶嗪和甲硫哒嗪的添加导致细胞MALT1活性之活化的大幅降低。(B)用甲哌啶嗪和甲硫哒嗪处理Jurkat T细胞以剂量依赖的方式防止刺激和MALT1依赖性切割RelB。用溶剂或2μM、5μM、10μM或20μM的甲哌啶嗪或甲硫哒嗪将Jurkat T细胞处理4小时和1小时,用MG132来稳定RelB切割片段(RelBΔ)。用P/I刺激细胞30分钟。通过Western印迹分析RelB和RelBΔ。印迹示出至少3个独立实验中的一个代表性实验。(C)为了分析PD对T细胞活化的抑制影响,在存在或不存在5μM和10μM的甲哌啶嗪或甲硫哒嗪的条件下,在P/I或抗CD3/CD28刺激20小时之后,通过ELISA测量Jurkat T细胞的IL-2分泌。两种化合物都导致T细胞活化之后降低的胞外IL-2水平。(D)PD化合物对原代鼠CD4+T细胞之活化的影响。用甲哌啶嗪或甲硫哒嗪预处理3小时并且用抗CD3/CD28诱导4小时之后,使用定量PCR来测定IL-2 mRNA水平。与经溶剂处理的对照细胞相比,经化合物处理的细胞中IL-2 mRNA水平显著降低。因此,用两种化合物处理细胞和用抗CD3/CD28抗体进行后续T细胞活化20小时导致较低水平的分泌的IL-2。图(在B至D中)示出至少3个独立实验的平均值。误差棒表示SD。(E)用5μM和10μM的甲哌啶嗪和甲硫哒嗪对来自两个供体的原代人PBMC进行处理3小时,然后用抗CD3/CD28诱导20小时。在两种化合物的存在下,在所有供体中胞外IL-2水平剂量依赖地降低。
图5:在ABC-DLBCL细胞中,PD处理减弱MALT1活性和后续的底物切割。(A)在用甲哌啶嗪和甲硫哒嗪孵育4小时之后,分析DLBCL中的细胞MALT1活性。通过基于抗体的沉淀分离MALT1,在检测释放之AMC荧光团的荧光发射的读板器中测定其蛋白水解活性。两种化合物以剂量依赖的方式抑制ABC-DLBCL细胞的MALT1蛋白酶活性,根据细胞系或PD而存在一些变化。图示出至少三个独立实验的平均值,误差棒示出SD。(B)用甲哌啶嗪和甲硫哒嗪处理DLBCL细胞可以防止BCL10以剂量依赖的方式进行组成性MALT1依赖性切割。用不同剂量的化合物将细胞处理20小时,通过Western印迹分析BCL10和切割产物BCL10Δ5的存在。数据代表至少3个独立实验。
图6:在ABC-DLBCL细胞中,甲哌啶嗪处理减弱NF-κB靶基因结合和表达。(A)用10μM和20μM的甲哌啶嗪将ABC-DLBCL细胞处理20小时,然后通过EMSA分析NF-κB DNA结合。在所有四个细胞系中,NF-κB靶基因结合都被减弱。因此,用甲哌啶嗪处理降低了靶向BCL-XL和c-FLIP-L的抗凋亡NF-κB的蛋白质水平。数据代表三个独立实验。(B)为了测定对NF-κB靶基因表达的作用,用甲哌啶嗪将ABC-DLBCL和GCB-DLBCL对照细胞处理20小时,通过ELISA分析组成性分泌细胞因子IL-6和IL-10的水平。ABC细胞系中细胞的处理导致IL-6和IL-10分泌降低约50%。为了解释在单个细胞系中细胞IL-6和IL-10分泌的显著变化,用两种不同的标度举例说明IL的量。图示出至少三个独立实验的平均值,误差棒表示SD。
图7:PD对ABC-DLBCL细胞具有选择性毒性。(A)至(D)为了测试PD对ABC-DLBCL细胞之活力的影响,用所述浓度的甲哌啶嗪或甲硫哒嗪(单次处理)处理四种不同的ABC细胞系和作为对照细胞的三种GCB-DLBCL细胞系BJAB、Su-DHL-6和Su-DHL-4。然后在两天后用MTT细胞毒性测试分析细胞的活力(A和C)或在4天后用细胞计数分析细胞的活力(B和D)。在ABC-DLBCL细胞系中,两种化合物都可以促进细胞活力的降低,而不会显著影响GCB-DLBCL细胞。(E)在甲哌啶嗪处理之后,分析ABC-DLBCL细胞系中的凋亡。用15μM甲哌啶嗪将五种ABC-DLBCL细胞系和两种GCB-DLBCL细胞系处理5天。通过FACS分析将凋亡细胞鉴定为膜联蛋白V-PE阳性细胞和7-AAD阴性细胞。虽然在GCB-DLBCL对照细胞系中凋亡未增加,但是在全部ABC-DLBCL细胞系中都检测到凋亡细胞群体10%至25%的增加。数据(在B和D中)是三个独立实验的平均值。图(在A、C和E中)示出至少3个独立实验的平均值,并且误差棒表示SD。
图8:在体内的ABC-DLBCL细胞系OCI-Ly10中,甲哌啶嗪和甲硫哒嗪干扰生长并且诱导凋亡。(A)在第0天将重悬浮在基质胶(BD)中的OCI-Ly10细胞或Su-DHL-6细胞移植到NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ(NSG)小鼠侧面。通过卡尺测量测定肿瘤大小。在移植后24小时开始溶剂、甲哌啶嗪(300μg/天)或甲硫哒嗪(400μg/天)腹膜内施用于每组的3只独立的小鼠,并在每24小时的整个处理期间持续给药。两种PD选择性地减弱ABC-DLBCL细胞系OCI-Ly10的生长。采用双因素方差检验进行统计分析,导致第16天至第22天的高度显著的p值为<0.0001。(B)吩噻嗪在体内增强OCI-Ly10细胞凋亡,但不增强Su-DHL-6细胞凋亡。在处理22天之后,通过TUNEL染色对肿瘤部分测定凋亡。图示出有代表性的肿瘤部分的染色。(C)甲哌啶嗪和甲硫哒嗪在OCI-Ly10肿瘤中抑制RelB切割。在22天之后通过Western印迹在OCl-Ly10肿瘤样品的提取物中检测到了RelB和MALT1依赖性切割产物RelBΔ的表达。印迹示出来自用溶剂、甲哌啶嗪或甲硫哒嗪处理之小鼠的结果,各自示出三个独立的样品。
图9:MALT1的抑制谱提示与拟南芥间胱天蛋白酶的高度相似性。类似于AtMC4和AtMC9,100μM的天门冬氨酰蛋白酶抑制剂胃蛋白酶抑制剂A和丝氨酸蛋白酶抑制剂抑肽酶(5μg/ml)都不能抑制MALT1蛋白水解活性。胰凝乳蛋白酶抑制剂(100μM)和抗蛋白酶(1μM)可以大幅抑制MALT1和间胱天蛋白酶,亮抑酶肽(1μM)对AtMC4/9具有较强的作用,而半胱氨酸蛋白酶抑制剂E-64不抑制MALT1,其对两种间胱天蛋白酶具有温和的作用。虽然TLCK(1μM)对间胱天蛋白酶具有轻微作用,但MALT1活性不受影响。高剂量(100μM)的DEVD四肽胱天蛋白酶抑制剂不抑制MALT1或AtMC4/9。
图10:MALT1HTS的参数。在初级筛选中,针对384孔形式中的170nM的GSTMALT1,测试了终浓度为10μM的ChemBioNet多样性文库的约18,000种小分子。在二级测定中使用5μM至50μM的不同剂量进一步验证具有最佳抑制潜力的所得300种命中(hit)。鉴定了15种二级命中,这相当于原始文库的约0.08%。
图11:(A)蛋白水解性CASP8测定的建立。用50μM的胱天蛋白酶底物Ac-DEVD-AMC测试不同量的活性重组CASP8(0.25μg、0.5μg和1μg)。根据GSTMALT1测定确定酶促活性。为了分析PD对CASP8的抑制作用,使用了250pg。数据代表两个独立实验。(B)在Ac-DEVD-CHO存在的条件下,针对Ac-DEVD-AMC的CASP8活性导致在50pM的浓度下酶促活性的几乎完全降低。图示出三个独立实验的平均值。误差棒表示SD。
图12:(A)普马嗪抑制细胞MALT1活性。在对细胞进行4小时的普马嗪处理之后,ABC-DLBCL中的组成性MALT1活性降低。(B)和(C)普马嗪减弱ABC-DLBCL细胞活力。与在细胞MALT1切割测定中获得的结果相一致,普马嗪对ABC-DLBCL细胞活力具有最温和的作用。(D)和(E)Malt1非活性异丙嗪不影响ABC-DLBCL活力。用10μM和20μM的异丙嗪将ABC-DLBCL和GCB-DLBCL细胞系处理4天,不显著减弱两种DLBCL亚组的活力。数据是三个独立实验的平均值。误差棒(在A、B和D中)表示SD。
图13:吩噻嗪衍生物和MALT1的结构活性关系(SAR)的阐明。示出了通过药物化学设计的不同吩噻嗪的化学结构和MALT1抑制潜力。这些化学结构落在上文所述的通式(I)的范围内。这证明,根据通式(I)的化合物是有效的MALT1抑制剂。
实施例对本发明进行了举例说明。
具体实施方式
实施例1-实验过程
细胞培养和试剂
将DLBCL细胞系培养在补充有20%FCS和100U/ml青霉素/链霉素的RPMI 1640培养基(Invitrogen)中,培养在具有20%人血浆、青霉素/链霉素和50μM β-巯基乙醇中的ABC系OCI-Ly10除外。根据DLBCL细胞系用10%FCS培养Jurkat T细胞。根据制造商(Axis-shield)用淋巴细胞分离剂(Lymphoprep)从经肝素处理(1000U/ml)的全血中分离人单核细胞(PBMC)。用T细胞特异性戴诺磁珠(Dynabeads,Invitrogen)进行鼠CD4+T细胞的分离。将原代细胞培养在含50μM β-巯基乙醇的Jurkat培养基中。通过添加佛波醇12-豆蔻酸酯13-乙酸酯(PMA;200ng/ml)和离子霉素(I;300ng/rrl)(均为Calbiochem)或者通过hCD3/hCD28和mIgG1/mIgG2a抗体(BD Biosciences)来启动对Jurkat T细胞、人PBMC和小鼠CD4+T细胞的刺激。将Z-VRPR-FMK(Alexis Biochemicals)、甲哌啶嗪乙酸酯(Chembridge)、盐酸普马嗪、盐酸甲硫哒嗪、盐酸异丙嗪(均为Sigma Aldrich)和所测试的所有其他PD(Chembridge或Sigma)溶解在DMSO中。
重组和内源MALT1切割测定
在感受态(competent)BL21 RIL大肠杆菌中产生GSTMALT1蛋白。在18℃下,在0.8的OD600下用50μM的异丙基-α-D-硫代半乳吡喃糖苷(IPTG)对蛋白质产生诱导16小时。收获细菌,并在裂解缓冲液(50mM HEPES,pH 7.5,10%甘油,0.1%Triton X-100,1mM二硫苏糖醇,150mM NaCl,2mM MgCl2,包括蛋白酶抑制剂)中通过超声裂解。使用谷胱甘肽FastTrap柱(GE Healthcare)通过液相色谱系统纯化GSTMALT1。就在384孔微孔板中的切割测定而言,使用200ng的蛋白质和50μM的BCL-10衍生的基底Ac-LRSR-AMC。在30℃孵育30分钟之后,使用Synergy 2微孔板读板器(Biotek)将所切割AMC的荧光测量1小时。将蛋白酶活性表示为相对荧光单位,其中将经DMSO处理的对照设置为100%,适当地计算经化合物处理之孔的荧光。因此,针对作为底物的Ac-DEVD-AMC并且分别针对50pg和250pg的蛋白质测定人重组CASP3(BioVision)和CASP8(Cayman Chemical)的切割。就内源MALT1蛋白酶而言,对DLBCL或Jurkat T细胞(5×106个细胞)不予处理、用抑制剂(分别为4小时和3小时)或P/I以及CD3/CD28进行处理并在4℃下在裂解缓冲剂中裂解。就免疫沉淀而言,向400μl的澄清裂解物中添加4μl的抗MALT1抗体(H-300,Santa Cruz Biotechnology)。在4℃孵育16小时之后,添加15μl的经PBS清洗的蛋白质G-琼脂糖珠(Roche),进一步将样品孵育1小时。用PBS将该珠清洗3次,重悬于40μl的切割测定缓冲液(50mM MES,pH 6.8,150mM NaCl,10%[重量/体积]蔗糖,0.1%[重量/体积]CHAPS,1M柠檬酸铵,10mM二硫苏糖醇)中,并转移到384孔微孔板中。添加肽底物Ac-LRSR-AMC至终浓度为20μM,根据重组GSTMALT1测定来测量活性。将所用的所有抑制剂溶解于DMSO中,并且用合适量的溶剂处理对照细胞。
对MALT1小分子抑制剂的高通量筛选(HTS)
在柏林的莱布尼茨分子药理学研究所(Leibniz Institute for MolecularPharmacology)(FMP)采用MALT1切割测定来筛选ChemBioNet文库的约18000种小分子35。在384孔非结合测定板(Corning)中,筛选体积为11μl,其中170nmol GSTMALT1针对10μM终浓度的化合物。在30℃,用50μM的Ac-LRSR-AMC底物将该测定进行20分钟。使用重组MALT1突变体C453A作为阴性对照,使用1nM的Z-VRPR-FMK肽作为介质抑制对照。通过标准的Z因子确定(约0.7)来确认测定的质量。就命中验证而言,用8种不同浓度(0.7μM至90.9μM)的化合物对来自初级筛选的具有最佳抑制作用的300种化合物测定两次。
通过实时RT-PCR进行RNA的定量
根据制造商的方案,通过用随机六聚体(random hexamer)和Superscript II(Invitrogen)进行反转录来用无DNA的RNA样品(RNeasy Mini Kit,Qiagen)进行cDNA的合成。在LC 480 Lightcycler系统(Roche)上使用LC 480 SybrGreen PCR混合物(Roche)来进行实时PCR。通过归一化至β-肌动蛋白管家基因来实现细胞因子RNA的定量。根据Pfaffl2001来计算相对表达比。使用下述引物:mIL-2正向5′-GAGTGCCAATTCGATGATGAG-3′(SEQ IDNO:1);mIL-2反向5′-AGGGCTTGTTGAGATGATGC-3′(SEQ ID NO:2);mβ-肌动蛋白正向5′CCTCTATGCCAACACAG TGC3′(SEQ ID NO:3);mβ-肌动蛋白反向5′-GTACTCCTGCTTGCTGATCC-3′(SEQ ID NO:4)。36
电泳迁移率变动测定(EMSA)、Western印迹和ELISA
如前文所述进行全细胞提取、Western印迹和EMSA9。所用抗体是BCL-XL(Cellsignaling)、MALT1(H300,B12)、BCL10(H197)、c-FLIP(Alexis Biochemicals)和β-肌动蛋白(I-19)。用不同剂量的PD将弥散性大B细胞淋巴瘤细胞处理20小时之后,对BCL10切割进行可视化。用甲哌啶嗪和甲硫哒嗪对Jurkat T细胞和原代人和小鼠细胞预处理3小时,然后进行T细胞受体刺激20小时,之后,根据制造商的方案进行人和鼠IL-2 ELISA(BenderMedSystems)。在对DLBCL细胞系进行20小时的抑制剂孵育之后,进行IL-6和IL-10 ELISA(Immunotools)。
活力、MTT和凋亡测定
与经DMSO处理的对照细胞相比,用剂量依赖的抑制剂处理2天之后,进行MTT(3-4,5-二甲基噻唑-2-基-2,5-联苯基四唑溴化物)细胞毒性测试,4天之后通过经台盼蓝(trypan blue)染色之细胞的细胞计数测定来分析DLBCL细胞系的活力。用μQuant微孔板分光光度计(Biotek)在λ=450nm处测量MTT细胞依赖性还原为甲。化合物处理5天之后,通过FACS分析(LSRII,BD)用7AAD-细胞(BD Pharmingen)的PE-膜联蛋白V染色测定凋亡率。使用FlowJo软件(Treestar)分析数据。
实施例2-MALT1对胱天蛋白酶(MALT1 paracaspase)表现出不同于人胱天蛋白酶的蛋白水解活性
为了筛选可以抑制MALT1蛋白酶活性的小分子量化合物,从大肠杆菌纯化重组GSTMALT1以建立适于高通量筛选(HTS)的体外蛋白酶切割测定。在30℃于50μM的四肽底物Ac-LRSR-AMC的存在下,将GSTMALT1孵育1小时,Ac-LRSR-AMC衍生自BCL10之C末端的MALT1切割部位7。通过测量荧光的增加来确定蛋白水解活性,所述荧光在切割之后发出并且伴随着荧光团AMC的释放(图1A和B)。MALT1催化的切割Ac-LRSR-AMC因荧光密度随时间的显著增加而明显。MALT1(同工型B)的对胱天蛋白酶结构域中保守半胱氨酸(C453A)的突变废除了MALT1催化活性(图1A)。类似于精氨酸-赖氨酸特异性间胱天蛋白酶,MALT1蛋白酶也对精氨酸残基之后的切割具有高偏好性。与该Z-VRPR-FMK相一致,其在最初设计为间胱天蛋白酶拮抗肽24,以及在低纳摩尔浓度也完全阻断MALT1切割活性,这强调了对胱天蛋白酶与植物间胱天蛋白酶的高度相似性(图1B和C)。相比之下,即使在皮摩尔浓度下也有效阻断CASP8活性(图11)的有效胱天蛋白酶抑制肽Ac-DEVD-CHO只少量地降低MALT1活性,即使在以200μM的浓度使用时也是如此(图1D)。
胱天蛋白酶和MALT1的不同底物特异性强调了鉴定这样的小分子抑制剂的可能性:所述小分子抑制剂干扰MALT1依赖性促生存信号传导20,21而不妨碍胱天蛋白酶依赖性凋亡机制。因为MALT1对胱天蛋白酶是植物间胱天蛋白酶的唯一哺乳动物类似物(homologue)4,所以进一步表征了MALT1酶促活性和底物偏好性。在蛋白酶抑制剂的存在下测定MALT1切割(图1E),并且如图9中总结的,将所述作用与对于植物间胱天蛋白酶AtMC4和AtMC9所得的抑制谱相比较5。正如AtMC4和AtMC9,天门冬氨酰蛋白酶抑制剂胃蛋白酶抑制剂A(100μM)和丝氨酸蛋白酶抑制剂抑肽酶(5μg/ml)都不大幅抑制MALT1活性。而广谱丝氨酸/半胱氨酸蛋白酶抑制剂胰凝乳蛋白酶抑制剂(100μM)和抗蛋白酶(1μM)将MALT1和AtMC4/9抑制至类似的程度,亮抑酶肽(1μM)对植物间胱天蛋白酶的作用更强。有趣的是,示出对AtMC4(而非AtMC9)具有温和作用的半胱氨酸蛋白酶抑制剂E-64(100μM)不抑制MALT1。相比之下,大幅抑制AtMC9的丝氨酸/半胱氨酸蛋白酶抑制剂TLCK(1μM)和弱得多的AtMC4仅温和地影响MALT1活性。如所期望的,四肽胱天蛋白酶抑制剂不抑制MALT1或AtMC4/9活性。总之,底物特异性和抑制谱表明MALT1对胱天蛋白酶与植物间胱天蛋白酶AtMC4/9之间的高相似性。
实施例3-吩噻嗪衍生物作为选择性MALT1蛋白酶抑制剂的鉴定
为了鉴定MALT1蛋白酶的小分子抑制剂,如图10所述采用测定形式筛选ChemBioNet采集物中的约18,000种化合物。在10μM的每种化合物的存在下,在20分钟的测定时间内,通过测量384个半孔(half-well)形式中AMC荧光的增加来进行初级筛选。300种初级命中示出抑制潜力,并且从中选择用于二级命中验证,所述二级命中验证以相同形式用增加剂量的每种化合物(0.7μM至90.9μM)进行两次。所述验证得出15种初级命中,这相当于初级筛选的约0.08%。
在研究15种初级命中之结构时,注意到效率和选择性最高的3种化合物(图2A:化合物A、B和C)是三环吩噻嗪的衍生物,所述三环吩噻嗪含有通过内环中的氮原子和硫原子连接的两个外部苯环。还发现抑制剂D中的杂环核心表现出与吩噻嗪的高结构相似性,而氮被碳所替代。这些初始结果表明,某些吩噻嗪衍生物(PD)可以充当MALT1抑制剂。为了验证MALT1抑制并为了评价特异性,测试四种经鉴定的PD的抑制MALT1和CASP8活性。在50μM,四种物质全都以剂量依赖的方式将MALT1蛋白酶活性降低至低于10%(图2B)。相比之下,CASP8活性只适中地影响50μM的最高抑制剂浓度,表明四种PD选择性地作用于MALT1。还测试了不含任何修饰的吩噻嗪支架,发现其以剂量依赖的方式抑制MALT1活性(图2C)。特别地,我们的初始结果表明,只有化合物A的修饰似乎显著地提高吩噻嗪骨架对MALT1的抑制潜力。有趣的是,化合物A对应于已知的药物甲哌啶嗪(之前的商标名为Pacatal),其已被用作镇静剂25。这些结果表明吩噻嗪可以是作为选择性MALT1抑制剂的有前景的候选物。
实施例4-充当有效且选择性的MALT1对胱天蛋白酶抑制剂的甲哌啶嗪、甲硫哒嗪和普马嗪
获得了甲哌啶嗪以及25种其他市售PD以测试其抑制潜力。而大多数化合物(12至26)没有或只有非常弱的抑制潜力(IC50>20μM),8种化合物(4至11)抑制MALT1活性,其IC50约为5μM至20μM。只有三种PD具有低于5μM的IC50。因此,只有一小亚组的PD能够有效地抑制MALT1。这三种最有效的化合物代表普马嗪、甲硫哒嗪和甲哌啶嗪,在筛选中首先鉴定了甲哌啶嗪(图3A)。为了定义抑制潜力,测定了每种化合物对重组全长(FL)GSTMALT1和酶促活性的截短的MALT1蛋白质(其包括对胱天蛋白酶的氨基酸和C末端Ig样(Ig3)结构域(第325位至第760位))的精确IC50值(图3B)。甲哌啶嗪在抑制GSTMALT1FL和GSTMALT1325-760中最有效,其IC50值分别为0.83μM和0.42μM。甲硫哒嗪和普马嗪也示出GSTMALT1 FL和GSTMALT1 325-760的剂量依赖抑制,但IC50值比甲哌啶嗪低约4(GSTMALT1 FL)或8(GSTMALT1 325-760)倍。相比之下,仍用于治疗某些精神疾病并且与这三种活性PD高度相关的药物异丙嗪在高达20μM的浓度下不引起任何显著的MALT1抑制。这些结果表明在MALT抑制中的高度特异性,即使在PD组中也是如此。
为了测试作用方式,基于荧光MALT1切割测定在米氏动力学中甲哌啶嗪的作用(图3C)。GSTMALT1 FL表现出约170RFU/分钟的VMAX,并且米氏常数(KM)计算为约48μM,其处于之前已测定的范围内(Hachmann等,2012)。IC50(1μM)左右浓度的甲哌啶嗪的添加大幅降低VMAX至约58RFU/分钟,而48μM的KM未被改变。甲哌啶嗪和其他吩噻嗪不含反应性基团。但是,为了确定甲哌啶嗪充当非共价可逆抑制剂,进行了使用连接至谷胱甘肽琼脂糖珠的GSTMALT1的洗出实验(wash-out experiment)(图3D)。此外,甲哌啶嗪抑制MALT1切割活性,但清洗GSTMALT1珠的数次循环导致抑制的完全丧失,即使在化合物的最高浓度(50μM)下也是如此。因此,甲哌啶嗪对MALT1酶促活性的作用揭示了由吩噻嗪引起的MALT1抑制的非竞争性形式和可逆形式。
接下来,测定PD对胱天蛋白酶的作用,在哺乳动物中,胱天蛋白酶是在结构上最接近于MALT1的相关物(Uren等,2000)。重要的是,三种PD都未显著抑制CASP3或CASP8活性,即使在浓度高至50μM时也是如此(图3E),反映出化合物作为MALT1抑制剂的选择性。
实施例5-在T细胞中,吩噻嗪抑制MALT1活性和IL-2诱导
在生理条件下,MALT1蛋白酶示出有助于T细胞应答。MALT1之活化腔中催化性半胱氨酸残基的突变防止应答于抗CD3/CD28共刺激的最佳IL-2产生(Duwel等,2009)。因此,确定了在T细胞中,PD对MALT1活性和IL-2产生的作用(图4)。在来自Jurkat T细胞之蛋白质的免疫沉淀(IP)之后,进行MALT1切割测定(图4A)。细胞不予处理或者用10μM的甲哌啶嗪或甲硫哒嗪孵育3小时,然后不予刺激或者用抗CD3/CD28进行刺激。在不存在刺激时,几乎检测不到MALT1蛋白酶活性,并且在CD3/CD28处理后30至60分钟达到峰值。在所有时间点于受刺激的Jurkat T细胞中,甲哌啶嗪或甲硫哒嗪的添加导致MALT1蛋白酶活性的大幅降低(图4A)。为了确认两种吩噻嗪都在细胞内抑制MALT1活性,监测在刺激Jurkat T细胞之后RelB的MALT1切割(图4B)。在P/I刺激之前,用蛋白酶体抑制剂MG132孵育Jurkat T细胞以防止不稳定的RelB截短物(truncation)降解时,可以检测到RelB切割产物RelBΔ(Hailfinger等,2011)。如因降低的RelBΔ水平和平行增加的全长RelB表达而明显的,甲哌啶嗪和甲硫哒嗪以剂量依赖的方式损伤RelB切割(图4B)。类似于重组MALT1的情况,甲哌啶嗪在抑制细胞MALT1切割活性中更有效,并且显著地将RelBΔ的表观浓度(appearance)降低为2至5μM,而在高于5μM时甲硫哒嗪有效。为了测定由PD引起的MALT1抑制对T细胞活化的作用,在存在或不存在甲哌啶嗪或甲硫哒嗪的条件下,在Jurkat T细胞的P/I或抗CD3/CD28刺激之后通过ELISA测量分泌的IL-2的量。两种化合物都导致T细胞活化后经PD处理之细胞的介质中IL-2水平的降低(图4C)。为了验证PD的抑制潜力在原代T细胞中也可检测到,对鼠CD4阳性Th1 T细胞进行分离和纯化,在存在或不存在5μM和10μM的甲哌啶嗪或甲硫哒嗪的情况下,在抗CD3/CD28共连接(co-ligation)之后,通过qPCR测量IL-2 mRNA诱导,并通过ELISA测量蛋白质水平(图4D)。IL-2 mRNA诱导和蛋白质表达二者均以剂量依赖的方式降低。最后,使用来自三个供体的原代人PBMC来评价抑制MALT1活性是否还促进原代人T细胞中IL-2产生的减少(图4E)。与之前的结果相符,在来自全部三个供体的PBMC中,甲哌啶嗪和甲硫哒嗪处理导致显著降低的IL-2分泌。
实施例6-吩噻嗪抑制ABC DLBCL细胞中MALT1活性和NF-κB靶基因的诱导
与MALT1底物A20和BCL10的组成性切割相符,MALT1蛋白酶活性增强,这是之前示出的所有ABC-DLBCL细胞的特性特征26。为了测定吩噻嗪对细胞MALT1活性的作用,用5μM或10μM的甲哌啶嗪、甲硫哒嗪和普马嗪将ABC-DLBCL细胞孵育4小时。进行抗MALT1 IP,通过向沉淀中添加底物AC-LRSR-AMC来测定MALT1蛋白酶活性。在ABC-DLBCL细胞中,三种PD都以剂量依赖的方式抑制MALT1蛋白酶活性(图5A)。尽管细胞MALT1活性的抑制根据各细胞系和化合物而改变,但甲哌啶嗪一般具有最强的作用,并且在10μM时,其在所有ABC-DLBCL细胞中导致MALT1活性降低至少75%。甲硫哒嗪也在所有ABC-DLBCL细胞系中抑制MALT1活性。但是在HBL1、U2932和TMD8中10μM甲硫哒嗪将MALT1抑制了多于80%,在OCI-Ly3和OCI-Ly10中仅观察到约50%的降低。普马嗪是细胞MALT1活性的最弱抑制剂。
接下来,评价了在ABC-DLBCL细胞中,由两种最强的化合物甲哌啶嗪和甲硫哒嗪引起的MALT1抑制是否还会防止已知MALT1底物BCL10的细胞切割(图5B)。MALT1切割BCL10的特定的C末端五个氨基酸,得到截短的切割产物(BCL10Δ5)。用增加剂量的每种化合物将ABC-DLBCL细胞处理20小时。事实上,用甲哌啶嗪或甲硫哒嗪的处理以剂量依赖的方式防止BCL10Δ5的检测。在ABC-DLBCL细胞中,MALT1活性有助于最佳NF-κB活化和靶基因表达20,21。因此,测定了在ABC-DLBCL细胞中,最强地影响MALT1活性的甲哌啶嗪是否还减弱组成性NF-κB DNA结合和随后的NF-κB靶基因表达(图6)。为此,用10μM和20μM的甲哌啶嗪将DLBCL细胞处理20小时,并通过EMSA分析NF-κB DNA结合(图6A)。在ABC-DLBCL细胞中,增加浓度的甲哌啶嗪导致降低的NF-κB靶DNA结合。相符地,甲哌啶嗪处理导致抗凋亡BCL-XL和FLIP-L蛋白质的剂量依赖性减少。为了进一步监测甲哌啶嗪对其他NF-κB依赖基因的作用,用10μM的甲哌啶嗪将ABC-DLBCL或GCB-DLBCL细胞处理20小时,通过ELISA测定细胞因子IL-6和IL-10的分泌(图6B)。但GCB-DLBCL细胞表达低量的IL-6或IL-10,ABC-DLBCL细胞分泌这两种细胞因子,虽然分泌的程度可变,这反映不同细胞系之间的异质性(heterogeneity)程度。重要的是,在所有ABC-DLBCL细胞(而非GCB-DLBCL细胞)中,甲哌啶嗪降低可溶性IL-6和IL-10的表达,表明其对NF-κB靶基因表达的直接作用。
实施例7-在ABC-DLBCL细胞中由吩噻嗪引起的选择性毒性和凋亡的诱导
由于三种PD在体外和体内有效地抑制MALT1蛋白酶活性,所以测试了这三种PD对ABC-DLBCL细胞活力的影响(图7)。采用三种GCB-DLBCL细胞系BJAB、Su-DHL-6和Su-DHL-4作为对照,其在之前示出其生长和存活不依赖于MALT1蛋白水解活性20。使用增加浓度的甲哌啶嗪、甲硫哒嗪和普马嗪孵育两天之后(单次处理),通过MTT测定测量细胞毒性作用(图7A、C和图12B)。在ABC-DLBCL细胞HBL1、OCI-Ly3、U2932和TMD8中,所有化合物都促进通过MTT反应测量的细胞活力降低,而不显著影响GCB-DLBCL细胞。此外,在处理4天之后,通过细胞计数来测定细胞活力(图7B、D和图12C)。与MTT测定相符,PD还降低ABC-DLBCL活细胞的总数。此外,ABC-DLBCL细胞降低的活力显著得多,而GCB-DLBCL细胞仅轻微受损,即使在化合物的最高浓度下也是如此。与细胞MALT1切割测定中获得的结果相符(图11A),普马嗪一般地对ABC-DLBCL细胞的活力具有最温和的作用。为了进一步验证施用不同PD之后ABC-DLBCL细胞的活力降低与MALT1抑制相关,用异丙嗪处理DLBCL细胞(图12E)。尽管异丙嗪与普马嗪结构关系密切,但异丙嗪在高至20μM的浓度下也不抑制MALT1蛋白酶活性(图12D)。事实上,在处理4天之后,异丙嗪不显著抑制ABC或GCB-DLBCL细胞的活力,这提供甲哌啶嗪、甲硫哒嗪和普马嗪的细胞作用依赖于MALT1抑制的进一步证据。
最后,已经通过增强凋亡测定了作为最有效MALT1抑制剂的甲哌啶嗪是否影响ABC-DLBCL细胞的活力(图7D)。为此,用15μM的甲哌啶嗪将DLBCL细胞处理5天,并通过FACS将凋亡细胞鉴定为膜联蛋白V-PE阳性细胞和7-AAD阴性细胞。在所有ABC-DLBCL细胞中,甲哌啶嗪诱发了提高的凋亡率,而在两种GCB-DLBCL对照细胞中,凋亡未增加。因此,PD对ABC-DLBCL细胞具有选择性毒性,并且该毒性部分地由于受影响淋巴瘤细胞中增加的凋亡,揭示了甲哌啶嗪和结构相关化合物用于ABC-DLBCL治疗的潜在用途。
实施例8-甲哌啶嗪和甲硫哒嗪阻止ABC-DLBCL在体内的生长
吩噻嗪(尤其是甲硫哒嗪)在治疗精神疾病方面的长久历史及其药理学和毒理学的详细知识可以促进用于治疗诊断有ABC-DLBCL之患者的标示外使用(off-label use)。因此,测定了在鼠DLBCL异种肿瘤模型中甲哌啶嗪和甲硫哒嗪是否还可以对体内淋巴瘤生长发挥作用。出于该目的,向NOD/scid IL-2Rgnull(NSG)小鼠中注射作为皮下异种移植物的ABC-DLBCL细胞系OCI-Ly10和GCB-DLBCL细胞系Su-DHL-6(图8A)。将两种肿瘤细胞系同时移植到单个小鼠的相对的侧面。从注射后一天开始,通过腹膜内施用溶剂或者甲哌啶嗪(12mg/kg)或甲硫哒嗪(16mg/kg)来处理小鼠。在经对照处理的小鼠中,在移植的三周内,从两种DLBCL细胞系都长出大的肿瘤。甲哌啶嗪或甲硫哒嗪的每日施用大幅削弱ABC-DLBCL细胞系OCI-Ly10的扩张。相比之下,在相同的动物中,两种PD完全未能对GCB-DLBCL细胞系Su-DHL-6的进展发挥任何抑制作用。
为了确认甲哌啶嗪和甲硫哒嗪直接作用于肿瘤细胞,测定了肿瘤组织中凋亡的诱导。在治疗期结束时移除移植的肿瘤,并通过对肿瘤组织部分进行TUNEL染色以使凋亡细胞可视化(图8B)。与在体内的选择性毒性相符,甲哌啶嗪或甲硫哒嗪处理增加异种移植的ABC-DLBCL细胞系OCI-Ly10中凋亡细胞的数目,而在GCB-DLBCL细胞系Su-DHL-6中未观察到凋亡的诱导。此外,在异种移植的OCI-Ly10肿瘤样本中甲哌啶嗪和甲硫哒嗪处理之后,MALT1底物RelB的组成性切割减弱,揭示了在小鼠中化合物确实也是通过在肿瘤细胞中抑制MALT1活性起作用的(图8C)。因此,鼠肿瘤模型提供了如下证据:由吩噻嗪引起的MALT1抑制在体内选择性地杀伤MALT1依赖性DLBCL,并且示出已知化合物在ABC-DLBCL治疗中之使用的潜在治疗益处。
参考文献
1.Thome M.Multifunctional roles for MALT1 in T-cell activation.Naturereviews Immunology.2008;8(7):495-500.Prepublished on 2008/06/26 as DOI10.1038/nri2338.
2.Scheidereit C.IkappaB kinase complexes:gateways to NF-kappaBactivation and transcription.Oncogene.2006;25(51):6685-6705.Prepublished on2006/10/31 as DOI 10.1038/sj.onc.1209934.
3.Oeckinghaus A,Wegener E,Welteke V,et al.Malt1 ubiquitinationtriggers NF-kappaB signaling upon T-cell activation.The EMBO journal.2007;26(22):4634-4645.Prepublished on 2007/10/20 as DOI 10.1038/sj.emboj.7601897.
4.Uren AG,O′Rourke K,Aravind LA,et al.Identification of paracaspasesand metacaspases:two ancient families of caspase-like proteins,one of whichplays a key role in MALT Iymphoma.Molecular cell.2000;6(4):961-967.Prepublished on 2000/11/25 as DOI.
5.Vercammen D,van de Cotte B,De Jaeger G,et al.Type II metacaspasesAtmc4 and Atmc9 of Arabidopsis thaliana cleave substrates after arginine andIysine.The Journal of biological chemistry.2004;279(44):45329-45336.Prepublished on 2004/08/25 as DOI 10.1074/jbc.M406329200.
6.Coornaert B,Baens M,Heyninck K,et al.T cell antigen receptorstimulation induces MALT1 paracaspase-mediated cleavage of the NF-kappaBinhibitor A20.Nature immunology.2008;9(3):263-271.Prepublished on 2008/01/29as DOI 10.1038/ni1561.
7.Rebeaud F,Hailfinger S,Posevitz-Fejfar A,et al.The proteolyticactivity of the paracaspase MALT1 is key in T cell activation.Natureimmunology.2008;9(3):272-281.Prepublished on 2008/02/12 as DOI 10.1038/ni1568.
8.Staal J,Driege Y,Bekaert T,et al.T-cell receptor-induced JNKactivation requires proteolytic inactivation of CYLD by MALT1.The EMBOjournal.2011;30(9):1742-1752.Prepublished on 2011/03/31 as DOI 10.1038/emboj.2011.85.
9.Duwel M,Welteke V,Oeckinghaus A,et al.A20 negatively regulates Tcell receptor signaling to NF-kappaB by cleaving Malt1 ubiquitinchains.Journal of immunology.2009;182(12):7718-7728.Prepublished on 2009/06/06 as DOI 10.4049/jimmunol.0803313.
10.Ngo VN,Davis RE,Lamy L,et al.A loss-of-function RNA interferencescreen for molecular targets in cancer.Nature.2006;441(7089):106-110.Prepublished on 2006/03/31 as DOI 10.1038/nature04687.
11.Alizadeh AA,Eisen MB,Davis RE,et al.Distinct types of diffuselarge B-cell Iymphoma identified by gene expression profiling.Nature.2000;403(6769):503-511.Prepublished on 2000/02/17 as DOI 10.1038/35000501.
12.Rosenwald A,Staudt LM.Gene expression profiling of diffuse IargeB-cell Iymphoma.Leukemia&Iymphoma.2003;44 Suppl 3:S41-47.Prepublished on2004/06/19 as DOI.
13.Rosenwald A,Wright G,Chan WC,et al.The use of molecular profilingto predict survival after chemotherapy for diffuse Iarge-B-cell Iymphoma.TheNew England journal of medicine.2002;346(25):1937-1947.Prepublished on 2002/06/21 as DOI 10.1056/NEJMoa012914.
14.Savage KJ,Monti S,Kutok JL,et al.The molecular signature ofmediastinal Iarge B-cell Iymphoma differs from that of other diffuse Iarge B-cell Iymphomas and shares features with classical HodgkinIymphoma.Blood.2003;102(12):3871-3879.Prepublished on 2003/08/23 as DOI10.1182/blood-2003-06-1841.
15.Wright G,Tan B,Rosenwald A,Hurt EH,Wiestner A,Staudt LM.A geneexpression-based method to diagnose clinically distinct subgroups of diffuseIarge B cell Iymphoma.Proceedings of the National Academy of Sciences of theUnited States of America.2003;100(17):9991-9996.Prepublished on 2003/08/06 asDOI 10.1073/pnas.1732008100.
16.Staudt LM,Dave S.The biology of human Iymphoid malignanciesrevealed by gene expression profiling.Advances in immunology.2005;87:163-208.Prepublished on 2005/08/17 as DOI 10.1016/S0065-2776(05)87005-1.
17.Davis RE,Brown KD,Siebenlist U,Staudt LM.Constitutive nuclearfactor kappaB activity is required for survival of activated B cell-likediffuse Iarge B cell Iymphoma cells.The Joumal of experimental medicine.2001;194(12):1861-1874.Prepublished on 2001/12/19 as DOI.
18.Lenz G,Davis RE,Ngo VN,et al.Oncogenic CARD11 mutations in humandiffuse Iarge B cell Iymphoma.Science.2008;319(5870):1676-1679.Prepublishedon 2008/03/08 as DOI 10.1126/science.1153629.
19.Davis RE,Ngo VN,Lenz G,et al.Chronic active B-cell-receptorsignalling in diffuse Iarge B-cell Iymphoma.Nature.2010;463(7277):88-92.Prepublished on 2010/01/08 as DOI 10.1038/nature08638.
20.Ferch U,Kloo B,Gewies A,et al.Inhibition of MALT1 proteaseactivity is selectively toxic for activated B cell-like diffuse Iarge B cellIymphoma cells.The Journal of experimental medicine.2009;206(11):2313-2320.Prepublished on 2009/10/21 as DOI 10.1084/jem.20091167.
21.Hailfinger S,Lenz G,Ngo V,et al.Essential role of MALT1 proteaseactivity in activated B cell-tike diffuse Iarge B-cell Iymphoma.Proceedingsof the National Academy of Sciences of the United States of America.2009;106(47):19946-19951.Prepublished on 2009/11/10 as DOI 10.1073/pnas.0907511106.
22.Isaacson PG,Du MQ.MALT Iymphoma:from morphology tomolecules.Nature reviews Cancer.2004;4(8):644-653.Prepublished on 2004/08/03as DOI 10.1038/nrc1409.
23.Rosebeck S,Madden L,Jin X,et al.Cleavage of NIK by the API2-MALT1fusion oncoprotein leads to noncanonical NF-kappaB activation.Science.2011;331(6016):468-472.Prepublished on 2011/01/29 as DOI 10.1126/science.1198946.
24.Vercammen D,Belenghi B,van de Cotte B,et al.Serpin1 of Arabidopsisthaliana is a suicide inhibitor for metacaspase 9.Journal of molecularbiology.2006;364(4):625-636.Prepublished on 2006/10/10 as DOI 10.1016/j.jmb.2006.09.010.
25.Whittier JR,Klein DF,Levine G,Weiss D.Mepazine(pacatal):clinicaltrial with placebo control and psychological study.Psychopharmacologia.1960;1:280-287.Prepublished on 1960/06/23 as DOI.
26.Kloo B,Nagel D,Pfeifer M,et al.Critical role of PI3K signaling forNF-kappaB-dependent survival in a subset of activated B-cell-like diffuseIarge B-cell Iymphoma cells.Proceedings of the National Academy of Sciencesof the United States of America.2011;108(1):272-277.Prepublished on 2010/12/22 as DOI 10.1073/pnas.1008969108.
27.Su H,Bidere N,Zheng L,et al.Requirement for caspase-8 in NF-kappaBactivation by antigen receptor.Science.2005;307(5714):1465-1468.Prepublishedon 2005/03/05 as DOI 10.1126/science.1104765.
28.Choi JH,Yang YR,Lee SK,et al.Potential inhibition of PDK1/Aktsignaling by phenothiazines suppresses cancer cell proliferation andsurvival.Annals of the New York Academy of Sciences.2008;1138:393-403.Prepublished on 2008/10/08 as DOI 10.1196/annals.1414.041.
29.Rho SB,Kim BR,Kang S.A gene signature-based approach identifiesthioridazine as an inhibitor of phosphatidylinositol-3′-kinase(PI3K)/AKTpathway in ovarian cancer cells.Gynecologic oncology.2011;120(1):121-127.Prepublished on 2010/11/03 as DOI 10.1016/j.ygyno.2010.10.003.
30.Seeman P,Lee T,Chau-Wong M,Wong K.Antipsychotic drug doses andneuroleptic/dopamine receptors.Nature.1976;261(5562):717-719.Prepublished on1976/06/24 as DOI.
31.Sarwer-Foner GJ,Koranyi EK.The clinical investigation of pacatalin open psychiatric settings.Canadian Medical Association journal.1957;77(5):450-459.Prepublished on 1957/09/01 as DOI.
32.Zhelev Z,Ohba H,Bakalova R,et al.Phenothiazines suppressproliferation and induce apoptosis in cultured Ieukemic cells without anyinfluence on the viability of normal Iymphocytes.Phenothiazines andIeukemia.Cancer chemotherapy and pharmacology.2004;53(3):267-275.Prepublishedon 2003/12/10 as DOI 10.1007/s00280-003-0738-1.
33.van Soolingen D,Hernandez-Pando R,Orozco H,et al.The antipsychoticthioridazine shows promising therapeutic activity in a mouse model ofmultidrug-resistant tuberculosis.PloS one.2010;5(9).Prepublished on 2010/09/17 as DOI 10.1371/journal.pone.0012640.
34.Weisman JL,Liou AP,Shelat AA,Cohen FE,Guy RK,DeRisi JL.Searchingfor new antimalarial therapeutics amongst known drugs.Chemical biology & drugdesign.2006;67(6):409-416.Prepublished on 2006/08/03 as DOI 10.1111/j.1747-0285.2006.00391.x.
35.Lisurek M,Rupp B,Wichard J,et al.Design of chemical Iibraries withpotentially bioactive molecules applying a maximum common substructureconcept.Molecular diversity.2010;14(2):401-408.Prepublished on 2009/08/18 asDOI 10.1007/s11030-009-9187-z.
36.Yin M,Zhang L,Sun XM,Mao LF,Pan J.Lack of apoE causes alterationof cytokines expression in young mice liver.Molecular biology reports.2010;37(4):2049-2054.Prepublished on 2009/08/01 as DOI 10.1007/s11033-009-9660-x.
Claims (14)
1.具有通式(I)的化合物:
其中:
X是N;
Y是S;
()z是C1-C5直链烷基链;
A是NR3R4、OR5或HET;
R1和R2在每次出现时独立地选自-H、-CH3、-OH、-OCH3、-SCH3、-F、-Cl以及-CF3;
R3、R4和R5是H或C1-C5直链或支链烷基,并且
HET是5、6或7元杂环,其中所述环原子可以是C、O或N,所述环可以是饱和的或芳族的,并且所述环可以被H或C1-C5直链或支链烷基取代;或者
通式(I)的可药用盐、对映体、非对映体、外消旋混合物、无定形形式或非溶剂化形式
在制备用于治疗MALT1依赖性免疫疾病的药物组合物中的用途。
2.权利要求1的用途,其中:
X是N;
Y是S;
()z是直链的C1-C5烷基链;
R1是-H;以及
R2是-H或-SCH3。
3.权利要求1或2的用途,其中,A是HET,并且HET是插入有NR3的5元至7元碳环。
4.权利要求1或2的用途,其中,A是NR3R4,并且R3是H或CH3,R4是-CH3。
5.权利要求1或2的用途,其中,A是NR3R4,其中R3是CH3,R4是-CH3、-C2H5或C3-C5直链烷基链,其链可以插入有O、N或S并且其与()z的碳原子形成饱和环。
6.权利要求5的用途,其中所述饱和环是插入有N的5元至7元碳环。
7.权利要求1或2的用途,其中,A是HET,并且HET是N-甲基哌啶-2-基或N-甲基哌啶-3-基。
8.权利要求1或2的用途,其中,
(a)Z=3,A是NR3R4,并且R3和R4是-CH3;
(b)Z=1并且A是N-甲基哌啶-3-基;或
(c)Z=2并且A是N-甲基哌啶-2-基。
9.权利要求1或2的用途,其中所述化合物是:
10.权利要求1或2的用途,其中MALT1依赖性免疫疾病是变应性炎症。
11.权利要求1或2的用途,其中MALT1依赖性免疫疾病是免疫系统的超敏感性,或慢性炎症。
12.权利要求11的用途,其中MALT1依赖性免疫疾病是变应反应或哮喘。
13.权利要求1或2的用途,其中MALT1依赖性免疫疾病是自身免疫病。
14.权利要求13的用途,其中MALT1依赖性免疫疾病选自多发性硬化、炎性肠病、红斑狼疮、银屑病、慢性阻塞性肺病、类风湿性关节炎或银屑病关节炎。
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Publication number | Priority date | Publication date | Assignee | Title |
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Families Citing this family (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014086478A1 (en) * | 2012-12-03 | 2014-06-12 | Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) | Inhibitors of malt1 protease |
EP3013818B1 (en) * | 2013-06-26 | 2021-12-29 | Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) | (s)-enantiomer of mepazine as paracaspase (malt1) inhibitor for treating cancer |
CN103709122B (zh) * | 2013-11-29 | 2016-05-25 | 四川大学 | 用于治疗的抗肿瘤和抗真菌化合物 |
US9198916B1 (en) | 2014-05-27 | 2015-12-01 | B&G Partners, Llc | Compounds and methods for treating tumors |
JPWO2017057695A1 (ja) * | 2015-09-30 | 2018-07-19 | 東レ株式会社 | ジフェニルピラゾール誘導体及びその医薬用途 |
US10103402B2 (en) * | 2015-12-01 | 2018-10-16 | University Of Kentucky Research Foundation | Liquid phenothiazine catholytes for non-aqueous redox flow batteries |
MX2018016339A (es) * | 2016-06-23 | 2019-09-04 | Bioimics Ab | Compuestos heterociclicos antiinfecciosos y sus usos. |
AU2017302182B2 (en) | 2016-07-29 | 2021-11-04 | Lupin Limited | Substituted thiazolo-pyridine compounds as MALT1 inhibitors |
RU2021113671A (ru) | 2016-07-29 | 2021-07-08 | Люпин Лимитед | Замещенные тиазолопиридиновые соединения в качестве ингибиторов malt1 |
US10662156B2 (en) | 2016-07-29 | 2020-05-26 | Toray Industries, Inc. | Guanidine derivative and medical use thereof |
JPWO2018159650A1 (ja) * | 2017-02-28 | 2019-12-19 | 東レ株式会社 | グアニジン誘導体及びその医薬用途 |
JP6958860B2 (ja) | 2017-11-07 | 2021-11-02 | 学校法人自治医科大学 | ミトコンドリアの機能障害の改善剤、及びミトコンドリアの機能障害に起因する疾患又は症状の予防又は治療薬、並びにそれらの用途 |
WO2023107721A1 (en) * | 2021-12-10 | 2023-06-15 | Rheos Medicines, Inc. | Methods for treating diseases using malt1 inhibitors |
CN116675682A (zh) * | 2023-05-23 | 2023-09-01 | 郑州大学 | 一种吩恶嗪类化合物及其制备方法和应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050192274A1 (en) * | 2000-11-06 | 2005-09-01 | Alexis Borisy | Combinations of drugs for the treatment of neoplastic disorders |
US20060009506A1 (en) * | 2004-07-09 | 2006-01-12 | Odyssey Thera, Inc. | Drugs for the treatment of neoplastic disorders |
Family Cites Families (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB774883A (en) * | 1954-06-21 | 1957-05-15 | Rhone Poulenc Sa | Phenthiazine derivatives |
US3156692A (en) * | 1958-07-23 | 1964-11-10 | Geigy Chem Corp | 5-basically substituted dibenzazepine compounds |
EP0138481B1 (en) * | 1983-10-05 | 1991-06-26 | Merck Frosst Canada Inc. | Leukotriene biosynthesis inhibitors |
GB9416571D0 (en) | 1994-08-16 | 1994-10-12 | Battelle Memorial Institute | Novel alkylamino derivatives as sigma 2 selective ligands |
US5679651A (en) * | 1995-06-05 | 1997-10-21 | Richardson; Bruce C. | Treatment for systemic lupus erythematosus |
US6333322B1 (en) | 1996-03-13 | 2001-12-25 | Eisai Co., Ltd. | Nitrogen-containing tricyclic compounds and drugs containing the same |
US5674329A (en) | 1996-04-26 | 1997-10-07 | General Electric Company | Adhesive tape covered laser shock peening |
US20030229082A1 (en) * | 2002-01-16 | 2003-12-11 | The Regents Of The University Of California | Inhibition of RNA function |
JP2005538065A (ja) * | 2002-06-17 | 2005-12-15 | フイラデルフイア・ヘルス・アンド・エデユケーシヨン・コーポレーシヨン | セロトニンファミリーの受容体および血液脳関門に関する免疫調節および細胞過程への影響 |
AU2005237208B2 (en) | 2004-04-30 | 2009-12-03 | Bkg Pharma Aps | Treatment of infectious diseases |
EP1948179A1 (en) | 2005-11-11 | 2008-07-30 | Boehringer Ingelheim International GmbH | Quinazoline derivatives for the treatment of cancer diseases |
AU2007290386A1 (en) * | 2006-09-01 | 2008-03-06 | Immune Control, Inc. | Novel compositions and methods for treatment of diseases related to activated lymphocytes |
GB0701170D0 (en) * | 2007-01-22 | 2007-02-28 | Imp Innovations Ltd | Compositions and uses thereof |
EP2247293A2 (en) | 2008-02-29 | 2010-11-10 | Immune Control, Inc. | Novel compositions and methods for treatment of diseases related to activated lymphocytes |
EP3013818B1 (en) | 2013-06-26 | 2021-12-29 | Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) | (s)-enantiomer of mepazine as paracaspase (malt1) inhibitor for treating cancer |
-
2012
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- 2012-08-01 CN CN201811390121.6A patent/CN109999041A/zh active Pending
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050192274A1 (en) * | 2000-11-06 | 2005-09-01 | Alexis Borisy | Combinations of drugs for the treatment of neoplastic disorders |
US20060009506A1 (en) * | 2004-07-09 | 2006-01-12 | Odyssey Thera, Inc. | Drugs for the treatment of neoplastic disorders |
Non-Patent Citations (1)
Title |
---|
ZHIVKO ZHELEV等: "Phenothiazines suppress proliferation and induce apoptosis in cultured leukemic cells without any influence on the viability of normal lymphocytes", 《CANCER CHEMOTHER PHARMACOL》 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113264903A (zh) * | 2021-05-27 | 2021-08-17 | 郑州大学 | 一种吩噻嗪类化合物及其制备方法和应用 |
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