CN115322208A - 一类新型的2-氨基噻唑类衍生物及其制备方法和医药用途 - Google Patents
一类新型的2-氨基噻唑类衍生物及其制备方法和医药用途 Download PDFInfo
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Abstract
本发明公开一类新型的2‑氨基噻唑类衍生物及其制备方法和医药用途,所述化合物如式I所示,还公开了含有所述化合物的药用组合物,以及所述化合物或其药用盐或含有其的组合物在制备治疗阿尔茨海默病及相关痴呆症状的药物中的应用。
Description
技术领域
本发明涉及药物化学领域,具体涉及一类具有β-淀粉样前体蛋白裂解酶1(BACE-1)抑制活性、抗痴呆活性的2-氨基噻唑类衍生物或其可药用的盐、制备方法及用途。
背景技术
阿尔茨海默病(Alzheimer's disease,AD)是老年人中最常见的痴呆类型,是一种慢性神经退行性疾病,导致认知功能的进行性丧失和日常功能的下降。在解释AD发病的假说中,Aβ级联假说一直占主导地位。淀粉样蛋白前体蛋白(Amyloid Precursor Protein,APP)是一种跨膜蛋白,存在于大脑神经元和神经胶质细胞中。在病理条件下,APP经β-分泌酶切割形成APP-β,随后再经γ分泌酶切割形成Aβ42单体片段。Aβ42是老年斑形成最早的形式,细胞外环境中Aβ42的积累会造成毒性的β-淀粉样斑块沉积。而BACE-1是Aβ产生过程中的限速酶。本发明发现了一类具有BACE-1抑制活性的2-氨基噻唑类化合物,在AD治疗方面具有很大的应用潜力。
发明内容
本发明的一个目的在于提供一种通式I所示的2-氨基噻唑类化合物或其可药用的盐。
本发明的另一个目的在于提供一种通式I所示的2-氨基噻唑类化合物的制备方法。
本发明的再一个目的在于提供一种药用组合物,其包含治疗有效量的选自通式I所示的2-氨基噻唑类化合物、其可药用的盐。
本发明的再一个目的在于提供一种BACE-1抑制剂,其包含选自通式I所示的2-氨基噻唑类化合物、其可药用的盐。
本发明的再一个目的在于提供通式I所示的2-氨基噻唑类化合物、其可药用的盐、含有其的药物组合物在治疗痴呆症状中的应用。其中,所述痴呆症状属于但不限于阿尔茨海默病、额-颞叶痴呆、克-雅病、路易体痴呆、帕金森病、或亨廷顿病等中枢神经***变性疾病。
技术方案:
基于上述目的,本发明所述的一种通式I所示的2-氨基噻唑类化合物、异构体或其可药用的盐:
其中:
Z为H或C1~C3烷基;
X为O或C或S;
Y为O或C或S;
R1,R3和R4为H或卤素;
R2为取代或未取代的-N(H)-C(O)-R5,或取代或未取代的-N(H)-S(O2)-R6,R5或R6独立的选自C6~C10芳基或C2~C10杂芳基;所述取代基选自H、卤素、-CN,-NO2、-OH,-COOH、-NH2、C1~C6烷基、C1~C6烷氧基、C1~C6卤代烷基、-N(H)-C(O)-C1~C3烷基或C1~C6烷氨基。
在一些实施例中,Z为H或甲基或乙基。
在一些实施例中,R2为取代或未取代的-N(H)-C(O)-R5,或取代或未取代的-N(H)-S(O2)-R6,R5或R6独立的选自苯、吡啶、吡嗪、噻吩、吡唑、噻唑、嘧啶、萘、呋喃、吡咯、茚、喹啉或吲哚;所述取代基选自H、卤素、-CN,-NO2、-OH,-COOH、-NH2、C1~C4烷基、C1~C4烷氧基、C1~C4卤代烷基、-N(H)-C(O)-C1~C4烷基或C1~C4烷氨基;在一些具体的实施例中,所述的取代基选自H、卤素、-CH3、-CN、-NO2、-CF3、-OCH3、乙基、正丙基、异丙基、叔丁基或-N(H)-C(O)-CH3。
在一些实施例中,Z为H或甲基;X为O或C;Y为O或C;R1,R3和R4为H;R2为取代或未取代的-N(H)-C(O)-R5,或取代或未取代的-N(H)-S(O2)-R6,R5或R6独立的选自苯、吡啶、吡嗪或萘,所述取代基选自:H、卤素、-CH3、-CN、,-NO2、-CF3、-OCH3、乙基、正丙基、异丙基、叔丁基或-N(H)-C(O)-CH3。
在一些实施例中,Z为H或甲基;X为O或C;Y为O或C;R1,R3和R4为H;R2为取代或未取代的-N(H)-C(O)-R5或取代或未取代的-N(H)-S(O2)-R6,R5或R6独立的选自苯、吡啶、吡嗪或萘,所述取代基选自:H、卤素、-CH3、-CN、-OCH3、异丙基、叔丁基或-N(H)-C(O)-CH3。
在一些具体的实施例中,本发明提供如下所述化合物的任一种,或其异构体或可药用的盐:
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氯甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-甲氧基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-甲基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-3,5-二氯甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-甲氧基吡嗪-2-甲酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-3-氯-5-甲基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)苯酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-氟苯酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4氟苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4氯苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4溴苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-(叔丁基)苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-(甲氧基)苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-甲基苯磺酰胺;
N-(4-(N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)氨磺酰)苯基)乙酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)萘-1-磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)萘-2-磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-2,4,6-三异丙基苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氯甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲氧基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-3,5-二氯甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲氧基吡嗪-2-甲酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-3-氯-5-甲基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)苯酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-氟苯酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氟苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氯苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4溴苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-(叔丁基)苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-(甲氧基)苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-甲基苯磺酰胺;
N-(4-(N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)氨磺酰)苯基)乙酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)萘-1-磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)萘-2-磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-2,4,6-三异丙基苯磺酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氯甲基吡啶酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲氧基甲基吡啶酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲基甲基吡啶酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)苯磺酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氟苯磺酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氯苯磺酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-(甲氧基)苯磺酰胺;
(R)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺;
(S)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺;
(R)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺;
(S)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺;
(R)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺;
(S)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺。
进一步地,所述通式I所示的化合物的制备方法,其特征在于,包括如下步骤:
在一些具体的实例中:
(1)色满酮1经硝化反应的到硝基色满酮中间体2,随后经Witting反应得到烯类中间体3,中间体3在硫氰酸银和碘单质的作用下环合,随后经氨基化得到母核中间体4,其经Boc保护得到中间体5,经过氢气还原得到中间体6,随后与不同取代的酰氯或磺酰氯进行缩合反应得到中间体7和8,最后经三氟乙酸脱Boc得到权利要求1所述的2-氨基噻唑类终产物。
(2)醛类化合物9经硼氢化钠还原得到醇类中间体10,随后与溴乙酸叔丁酯或2-溴丙酸叔丁酯反应生成中间体11,酸化后得到酸类中间体12,中间体12与N,O-二甲羟胺盐酸盐反应生成酰胺类中间体13,随后在叔丁基锂的催化下化合物得到母核中间体14,经Witting反应得到烯类中间体15,随后在硫氰酸银和碘的作用下环合得到中间体16,其经Boc保护得到中间体17,经过氢气还原得到中间体18,随后与不同取代的酰氯或磺酰氯进行缩合反应得到中间体19和20,最后经三氟乙酸脱Boc得到权利要求1所述的2-氨基噻唑类终产物。
本申请公开的化合物或其可药用的盐在制备BACE-1抑制剂药物中的应用也在本发明的保护范围内。
本申请公开的化合物或其可药用的盐在制备抗痴呆症状药物中的应用也在本发明的保护范围内。所述痴呆症状属于但不限于阿尔茨海默病、额-颞叶痴呆、克-雅病、路易体痴呆、帕金森病、或亨廷顿病等中枢神经***变性疾病。
本发明还公开了一种药物组合物,其包含化合物I及药学上可接受的辅料。
所述药物组合物在制备BACE-1抑制剂药物中的应用,以及在制备抗痴呆症状药物中的应用也在本发明的保护范围内。
有益效果:本申请所述的2-氨基噻唑类化合物为一类具有全新骨架结构的化合物,其可有效抑制BACE-1的活性。通过药理实验可见,所述2-氨基噻唑类化合物对于BACE-1靶点具有明显的抑制作用。其在AD治疗方面具有潜在的成药性开发前景。
除非另外说明,在说明书和权利要求中使用的以下术语具有下面讨论的含义:
“卤素”表示氟、氯、溴或碘,优选为氟、氯或溴。
“烷基”表示1-20个碳原子的饱和的脂烃基,包括直链和支链基团(本申请书中提到的数字范围,例如“1-6”,是指该基团,此时为烷基,可以含1个碳原子、2个碳原子、3个碳原子等,直至包括6个碳原子)。例如甲基、乙基、丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基、戊基等。C1~C6烷基是指具有1~6个碳原子的直链或支链的烷基,例如甲基、乙基、丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基、戊基、新戊基、己基等。
“卤代烷基”表示卤素取代的烷基,如本发明所述的烷基,它被一个或多个相同或不同的卤原子取代,例如-CH2Cl、-CF3、-CH2CF3、-CH2CCl3等。
“烷氧基”表示-O-(未取代的烷基)或-O-(未取代的环烷基)。代表性实例包括但不限于甲氧基、乙氧基、丙氧基、丁氧基等。
“烷氨基”表示-N-(未取代的烷基)或-N-(未取代的环烷基)。代表性实例包括但不限于甲氨基、乙氨基、丙氨基、丁氨基等。
“芳基”表示1至12个碳原子的全碳单环或稠合多环基团,具有完全共轭的π电子***。芳基的非限制性实例有苯基、萘基和蒽基。芳基可以是取代的或未取代的。当被取代时,取代基优选为一个或多个,更优选为一个、两个或三个,进而更优选为一个或两个。
“杂芳基”表示环中多达3-10个原子的稳定的单环或每个环中多达3-10个原子的双环碳环,其中至少一个环为芳香环且含有1-4个选自O、N和S的杂原子。未取代的杂芳基地非限制性实例有吡咯基、呋喃基、噻吩基、咪唑基、噁唑基、噻唑基、吡唑基、嘧啶基、喹啉基、异喹啉基、嘌呤基、吖啶基、咔唑基、噌啉基、喹噁啉基、吲哚基、苯并***基、苯并噻吩基、苯并呋喃基、异噁唑基、吲哚基、吡嗪基、哒嗪基、吡啶基、四唑基、三嗪基和咔唑基等。杂芳基可以是取代的或未取代的。当被取代时,取代基优选为一个或多个,更为优选为一个、两个或三个,进而更为优选一个或两个。
“取代或未取代”表示可以不被取代基取代,也可以被本发明所述的取代基任意取代。当为取代时要理解本领域普通技术人员可选择本发明化合物的取代基及取代型式而提供化学上稳定的并可通过本领域技术和方法自可以获得的原料可以合成的化合物。如果取代基自身被超过一个基团取代,应理解这些基团可在相同碳原子上或不同碳原子上,只要使结构稳定。
具体实施方式
以下通过实施例形式展示具体的实施方式,对本发明内容进行进一步的详细说明,但不应当将此理解为本发明上述主题的范围仅限于以下的实施例,凡是基于本发明上述内容在本领域内所能实现的技术均应属于本发明的内容。
实施例1
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺
(1)将2.0g(13.5mmol)苯并二氢吡喃-4-酮加至100mL两口瓶中,加入20mL浓硫酸,冰浴条件下搅拌10min将其溶解,然后将1.5g(14.85mmol)KNO3的硫酸溶液缓慢滴加至苯并二氢吡喃-4-酮的反应液中,继续搅拌反应2h后,将反应混合物倒入200mL冰水中继续搅拌30min,抽滤。将固体用水洗3次,干燥,甲醇重结晶得到中间体2。将甲基三苯基溴化膦(20.71mmol)加入到无水THF,0℃下,氩气保护,缓慢加入正丁基锂(18.64mmol),随后搅拌一个小时,加入溶于无水THF的中间体2(10.35mmol),升到室温,反应约一小时,之后用水淬灭反应,乙酸乙酯萃取,合并有机层,浓缩,用PE/EA=8:1柱层析得到中间体3。在0℃下将中间体3(7.85mmol)加入到碘(15.69mmol)和硫氰酸银(31.38mmol)的甲苯溶液中,室温搅拌过夜,抽滤,滤液用饱和亚硫酸钠洗,用乙酸乙酯萃取,合并有机层,随后将得到的有机层旋干,0℃下,将其溶于THF中并加入到NH3的乙醇溶液(2M,25mL),搅拌一个小时,随后在室温下搅拌2.5天。浓缩后用乙酸乙酯洗,抽滤,得中间体4,将其溶于THF中,加入三乙胺(28.27mmol)和二碳酸二叔丁酯(28.27mmol),室温反应2天,浓缩溶剂,直接用PE/EA=8:1柱层析得到中间体5。将中间体5(3.39mmol)加入到乙醇和饱和氯化铵溶液1:1的溶液(30mL)中,随后加入铁粉(6.79mmol),在87摄氏度下加热半个小时,趁热硅藻土过滤、乙酸乙酯萃取三次,合并有机层,浓缩溶剂后得到中间体6直接投下一步。将商业所得的吡啶-2-甲酸(0.67mmol)溶于20mL甲苯中,缓慢加入氯化亚砜(3.38mmol),并加入1滴DMF,在120度下反应1小时,冷却至室温后,浓缩溶剂得到吡啶-2甲酰氯备用。将吡啶-2甲酰氯(0.67mmol)溶于5mL的THF中,冰浴下将该溶液加入至中间体6(0.59mmol)的THF(5mL)中,加入吡啶(30μL),随后升温到室温并且搅拌反应30分钟,随后用10mL饱和碳酸氢钠溶液淬灭,用乙酸乙酯萃取三次。浓缩溶剂得中间体7,将中间体7(0.59mmol)溶于10mL的无水二氯甲烷中,加入10mL三氟乙酸,室温反应2小时,随后用饱和碳酸氢钠溶液调PH=9。加入乙酸乙酯萃取三次,合并有机相,浓缩后用DCM:MeOH=20:1柱层析得到2-氨基噻唑类终产物实施例I。
白色固体,该部分合成总产率5.6%。1H NMR(300MHz,CDCl3)δ9.90(s,1H),8.61(d,J=3.7Hz,1H),8.29(d,J=7.7Hz,1H),7.91(t,J=7.6Hz,1H),7.88(s,1H),7.54(d,J=7.9Hz,1H),7.51–7.42(m,1H),6.83(d,J=8.7Hz,1H),4.62(s,1H),4.35(d,J=11.4Hz,1H),4.166(s,1H),3.67(d,J=11.0Hz,1H),3.45(d,J=11.2Hz,1H),2.18(s,2H);HRMS(ESI)calcd for C17H17N4O2S[M+H]+341.1067,found 341.1067.
实施例2
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺
同实施例1的操作,黄色固体,产率4.2%。1H NMR(300MHz,DMSO-d6)δ10.68(s,1H),9.16(s,1H),8.55(s,1H),8.25(d,J=7.8Hz,1H),7.94(s,1H),7.62(d,J=7.7Hz,1H),6.74(d,J=8.5Hz,3H),4.33–4.13(m,2H),3.56–3.39(m,2H),1.98(d,J=38.7Hz,2H);13C NMR(101MHz,DMSO-d6)δ162.3,161.2,153.2,151.8,150.6,142.5,131.2,128.9,122.6,121.9,120.6,117.1,116.6,111.8,73.4,64.1,46.7,33.7;HRMS(ESI)calcd for C18H16N5O2S[M+H]+366.1019,found 366.1023.
实施例3
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺
同实施例1的操作,白色固体,产率6.5%。1HNMR(300MHz,CDCl3)δ9.68(s,1H),8.43(s,1H),8.32(dd,J=8.5,4.4Hz,1H),7.76(s,1H),7.57(dd,J=15.4,8.6Hz,2H),6.84(d,J=8.9Hz,1H),4.72(s,2H),4.39(d,J=11.3Hz,1H),4.20(s,1H),3.69(d,J=11.0Hz,1H),3.48(d,J=11.4Hz,1H),2.20(s,2H);13C NMR(101MHz,CDCl3)δ160.7,150.7,136.6,130.8,124.1,121.4,119.1,117.4,73.9,64.1,47.8,33.0;HRMS(ESI)calcd for C17H16FN4O2S[M+H]+359.0973,found 359.0979.
实施例4
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氯甲基吡啶酰胺
同实施例1的操作,白色固体,产率5.2%。1H NMR(400MHz,DMSO-d6)δ10.51(s,1H),8.76(s,1H),8.16(d,J=16.1Hz,2H),7.93(s,1H),7.62(d,J=6.8Hz,1H),6.77(s,2H),6.75(d,J=8.1Hz,1H),4.24(d,J=32.7Hz,2H),3.59–3.43(m,2H),2.01(d,J=53.0Hz,2H);13C NMR(101MHz,DMSO-d6)δ161.6,150.4,149.2,147.4,138.2,134.5,131.5,124.2,121.7,120.4,116.6,64.1,46.6,33.7;HRMS(ESI)calcd for C17H16ClN4O2S[M+H]+375.0677,found 375.0680.
实施例5
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-甲氧基甲基吡啶酰胺
同实施例1的操作,白色固体,产率6.3%。1H NMR(400MHz,DMSO-d6)δ10.31(s,1H),8.37(d,J=2.2Hz,1H),8.10(d,J=8.7Hz,1H),7.92(d,J=1.6Hz,1H),7.61(dd,J=9.5,4.2Hz,2H),7.23–6.74(m,2H),6.74(d,J=8.8Hz,1H),4.33–4.17(m,2H),3.93(s,3H),3.49(dd,J=37.5,9.6Hz,2H),2.13–1.88(m,2H);13CNMR(75MHz,DMSO-d6)δ162.2,158.1,150.1,143.0,136.8,131.8,128.6,124.0,121.4,120.0,116.6,64.1,56.4,46.6,33.7;HRMS(ESI)calcd for C18H19N4O3S[M+H]+371.1172,found 371.1177.
实施例6
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-甲基甲基吡啶酰胺
同实施例1的操作,白色固体,产率3.6%。1HNMR(400MHz,CDCl3)δ9.85(s,1H),8.40(s,1H),8.16(d,J=7.9Hz,1H),7.79(s,1H),7.68(d,J=7.9Hz,1H),7.54(dd,J=8.8,2.0Hz,1H),6.83(d,J=8.8Hz,1H),4.48(d,J=59.2Hz,2H),4.38(dd,J=7.2,4.3Hz,1H),4.23–4.15(m,1H),3.69(d,J=11.2Hz,1H),3.46(d,J=11.2Hz,1H),2.43(s,3H),2.21(d,J=14.0Hz,2H);13C NMR(75MHz,CDCl3)δ162.0,160.6,150.6,148.5,147.5,138.0,136.6,131.1,127.6,122.0,121.5,119.1,117.4,73.8,64.1,47.7,33.1,18.6;HRMS(ESI)calcdfor C18H19N4O2S[M+H]+355.1223,found 355.1228.
实施例7
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-3,5-二氯甲基吡啶酰胺
同实施例1的操作,白色固体,产率5.2%。1H NMR(400MHz,DMSO-d6)δ10.52(s,1H),8.70(s,1H),8.42(s,1H),7.67(d,J=1.4Hz,1H),7.52(d,J=8.7Hz,1H),6.75(d,J=8.8Hz,1H),6.63(s,2H),4.29(d,J=11.3Hz,1H),4.20(t,J=10.6Hz,1H),3.52(d,J=11.2Hz,1H),3.36(d,J=6.9Hz,1H),2.06(d,J=13.7Hz,1H),1.92(t,J=10.6Hz,1H);13CNMR(101MHz,DMSO-d6)δ162.4,150.3,146.3,138.3,132.6,131.6,129.3,121.0,119.5,116.8,73.7,64.1,46.9,33.7;HRMS(ESI)calcd for C17H15Cl2N4O2S[M+H]+409.0287,found409.0297.
实施例8
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-甲氧基吡嗪-2-甲酰胺
同实施例1的操作,白色固体,产率4.5%。1H NMR(400MHz,DMSO-d6)δ10.34(s,1H),8.88(t,J=9.4Hz,1H),8.38(d,J=1.2Hz,1H),7.92(d,J=2.5Hz,1H),7.57(dd,J=8.8,2.6Hz,1H),6.75(t,J=8.0Hz,3H),4.31–4.26(m,1H),4.19(dd,J=10.9,8.9Hz,1H),4.01(s,3H),3.52(d,J=11.3Hz,1H),3.42(d,J=7.0Hz,1H),2.08(dd,J=12.6,2.9Hz,1H),1.98–1.89(m,1H);13C NMR(101MHz,DMSO-d6)δ162.0,161.5,150.3,141.7,138.5,133.9,131.6,128.4,121.9,120.4,116.6,114.7,64.0,54.7,46.4,33.7;HRMS(ESI)calcd forC17H18N5O3S[M+H]+372.1125,found372.1131.
实施例9
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-3-氯-5-甲基甲基吡啶酰胺
同实施例1的操作,白色固体,产率4.5%。1HNMR(400MHz,CDCl3)δ9.90(s,1H),8.40(s,1H),7.66(d,J=7.7Hz,2H),7.52(d,J=8.1Hz,1H),6.84(d,J=8.7Hz,1H),4.86(s,2H),4.40(d,J=11.3Hz,1H),4.21(t,J=10.3Hz,1H),3.67(d,J=11.2Hz,1H),3.46(d,J=11.2Hz,1H),2.83(s,3H),2.28–2.10(m,2H);13C NMR(75MHz,CDCl3)δ162.7,160.8,150.8,145.1,144.2,140.5,137.6,134.1,130.9,127.7,121.8,119.4,117.4,73.8,64.1,47.7,32.7,20.8;HRMS(ESI)calcd for C18H18ClN4O2S[M+H]+389.0834,found 389.0841.
实施例10
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)苯酰胺
同实施例1的操作,白色固体,产率5.6%。1H NMR(400MHz,DMSO-d6)δ10.14(s,1H),7.95(s,2H),7.71(s,1H),7.54(d,J=23.6Hz,4H),6.74(d,J=7.2Hz,1H),6.56(s,2H),4.37–4.11(m,2H),3.60–3.47(m,2H),2.15–1.86(m,2H);13C NMR(101MHz,DMSO-d6)δ165.4,159.0,150.1,135.4,132.3,131.8,128.7,127.9,121.8,120.5,116.4,73.9,64.1,47.0,33.7;HRMS(ESI)calcd for C18H18N3O2S[M+H]+340.1114,found 340.1116.
实施例11
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-氟苯酰胺
同实施例1的操作,白色固体,产率5.3%。1H NMR(400MHz,DMSO-d6)δ10.15(s,1H),8.04(s,2H),7.69(s,1H),7.58(s,1H),7.35(s,2H),6.75(s,1H),6.61(s,2H),4.24(d,J=33.5Hz,2H),3.49(d,J=24.3Hz,2H),1.99(d,J=54.3Hz,2H);13CNMR(101MHz,DMSO-d6)δ164.3,150.1,132.2,130.7,129.1,121.9,120.5,116.5,115.8,115.5,73.8,64.1,46.9,33.7;HRMS(ESI)calcd for C18H17FN3O2S[M+H]+358.1020,found 358.1025.
实施例12
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)苯磺酰胺
同实施例1的操作,白色固体,产率4.5%。1HNMR(400MHz,CDCl3)δ7.69(d,J=5.4Hz,2H),7.63–7.36(m,3H),6.95(s,1H),6.12(d,J=7.2Hz,1H),5.77(s,1H),4.25(d,J=8.9Hz,1H),4.01(t,J=10.8Hz,1H),3.50(d,J=10.5Hz,1H),3.32(d,J=10.8Hz,1H),2.25(d,J=13.4Hz,1H),1.88(d,J=10.8Hz,1H);HRMS(ESI)calcd for C17H18N3O3S2[M+H]+376.0784,found 376.0792.
实施例13
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4氟苯磺酰胺
同实施例1的操作,白色固体,产率6.3%。1H NMR(300MHz,CDCl3)δ7.80–7.68(m,2H),7.15(t,J=8.6Hz,2H),6.93(d,J=2.5Hz,1H),6.17(d,J=8.6Hz,1H),5.77(dd,J=8.6,2.5Hz,1H),4.28(dd,J=8.1,3.5Hz,1H),4.05(t,J=11.4Hz,1H),3.53(d,J=11.1Hz,1H),3.36(d,J=11.3Hz,1H),2.27(d,J=14.5Hz,1H),1.85(t,J=11.3Hz,1H);HRMS(ESI)calcd for C17H17FN3O3S2[M+H]+394.0690,found394.0692.
实施例14
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4氯苯磺酰胺
同实施例1的操作,白色固体,产率5.2%。1HNMR(300MHz,CDCl3)δ7.62(d,J=7.6Hz,2H),7.41(d,J=7.4Hz,2H),6.92(s,1H),6.17(d,J=8.4Hz,1H),5.81(d,J=8.7Hz,1H),4.27(d,J=11.1Hz,1H),4.02(t,J=12.0Hz,1H),3.49(d,J=11.1Hz,1H),3.33(d,J=11.3Hz,1H),2.25(d,J=14.1Hz,1H),1.87(t,J=13.1Hz,1H);HRMS(ESI)calcd forC17H17ClN3O3S2[M+H]+410.0394,found 410.0402.
实施例15
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4溴苯磺酰胺
同实施例1的操作,白色固体,产率4.3%。1H NMR(400MHz,CDCl3)δ7.64–7.54(m,4H),7.28(s,1H),6.93(d,J=2.4Hz,1H),6.20(dd,J=8.5,4.3Hz,1H),5.88–5.78(m,1H),4.30(dd,J=7.9,3.7Hz,1H),4.05(t,J=11.8Hz,1H),3.56–3.48(m,1H),3.36(dd,J=11.2,4.2Hz,1H),2.28(d,J=12.9Hz,1H),1.89(t,J=12.9Hz,1H);HRMS(ESI)calcd forC17H17BrN3O3S2[M+H]+453.9889,found 453.9892.
实施例16
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-(叔丁基)苯磺酰胺
同实施例1的操作,白色固体,产率4.6%。1H NMR(300MHz,CDCl3)δ7.63–7.57(m,2H),7.46–7.39(m,2H),6.95(d,J=2.5Hz,1H),6.19(d,J=8.6Hz,1H),5.94(dd,J=8.7,2.5Hz,1H),4.27(dt,J=11.4,3.7Hz,1H),4.03(dd,J=11.6,10.1Hz,1H),3.45(d,J=11.3Hz,1H),3.30(d,J=11.3Hz,1H),2.25(d,J=15.4Hz,1H),1.93(t,J=11.1Hz,1H),1.32(s,9H);HRMS(ESI)calcd for C21H26N3O3S2[M+H]+432.1410,found 432.1411.
实施例17
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-(甲氧基)苯磺酰胺
同实施例1的操作,白色固体,产率3.8%。1HNMR(300MHz,DMSO-d6)δ7.60(d,J=8.9Hz,2H),7.04(d,J=8.9Hz,2H),7.00(d,J=2.6Hz,1H),6.79(dd,J=8.7,2.6Hz,1H),6.60(d,J=8.7Hz,1H),6.59(s,1H),4.28–4.04(m,2H),3.79(s,3H),3.43(s,2H),3.39(s,1H),3.07(d,J=11.2Hz,1H),1.90(ddd,J=20.2,14.0,7.2Hz,2H);13C NMR(101MHz,DMSO-d6)δ162.7,150.8,131.5,130.4,129.4,123.1,121.6,117.1,114.6,73.7,64.1,56.0,46.8,33.8;HRMS(ESI)calcd for C18H19N3O4S2[M+H]+406.0890,found 406.0897.
实施例18
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-甲基苯磺酰胺
同实施例1的操作,白色固体,产率4.5%。1HNMR(300MHz,CDCl3)δ7.57(d,J=8.3Hz,2H),7.22(d,J=8.0Hz,2H),6.94(d,J=2.5Hz,1H),6.21(d,J=8.7Hz,1H),5.99–5.92(m,1H),4.33–4.22(m,1H),4.04(t,J=10.7Hz,1H),3.49(d,J=10.8Hz,1H),3.32(d,J=11.3Hz,1H),2.40(s,3H),2.24(d,J=15.5Hz,2H);HRMS(ESI)calcd for C18H20N3O3S2[M+H]+390.0941,found 390.0947.
实施例19
N-(4-(N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)氨磺酰)苯基)乙酰胺
同实施例1的操作,白色固体,产率3.6%。1H NMR(300MHz,DMSO-d6)δ10.31(s,1H),9.71(s,1H),7.69(d,J=8.9Hz,2H),7.58(d,J=8.8Hz,2H),7.03(d,J=2.5Hz,1H),6.80(dd,J=8.7,2.6Hz,1H),6.61(d,J=8.7Hz,1H),4.28–4.07(m,2H),3.44(d,J=11.3Hz,1H),3.11(d,J=11.2Hz,1H),2.06(s,3H),2.00(d,J=16.1Hz,1H),1.86(t,J=10.3Hz,1H);HRMS(ESI)calcd for C19H21N4O4S2[M+H]+433.0999,found 433.1003.
实施例20
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)萘-1-磺酰胺
同实施例1的操作,白色固体,产率6.8%。1H NMR(300MHz,DMSO-d6)δ10.20(s,1H),8.67(d,J=8.2Hz,1H),8.20(d,J=8.2Hz,1H),8.09(dd,J=12.8,7.5Hz,2H),7.65(ddd,J=21.9,15.4,6.7Hz,3H),6.95(d,J=2.5Hz,1H),6.70(dd,J=8.7,2.5Hz,1H),6.53(s,1H),6.50(s,1H),4.23–3.98(m,2H),3.29(s,1H),2.85(d,J=11.3Hz,1H),1.97–1.71(m,2H);13C NMR(101MHz,DMSO-d6)δ150.5,134.9,134.6,134.1,130.4,130.0,129.4,128.4,128.0,127.3,124.9,122.2,120.7,117.0,64.1,46.7,33.9;HRMS(ESI)calcd forC21H20N3O3S2[M+H]+426.0941,found426.0947.
实施例21
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)萘-2-磺酰胺
同实施例1的操作,白色固体,产率6.2%。1HNMR(300MHz,DMSO-d6)δ9.94(s,1H),8.30(s,1H),8.05(dd,J=24.2,8.2Hz,3H),7.68(ddd,J=23.2,11.3,4.2Hz,3H),7.04(d,J=2.6Hz,1H),6.84(dd,J=8.8,2.6Hz,1H),6.58(d,J=8.7Hz,1H),6.57–6.36(m,1H),4.25–4.01(m,2H),3.31(d,J=11.4Hz,1H),2.91(d,J=11.0Hz,1H),2.02–1.74(m,2H);HRMS(ESI)calcd for C21H20N3O3S2[M+H]+426.0941,found 426.0946.
实施例22
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-2,4,6-三异丙基苯磺酰胺
同实施例1的操作,白色固体,产率4.6%。1HNMR(300MHz,CDCl3)δ10.40(s,1H),7.03(s,3H),6.21(d,J=8.6Hz,1H),6.05(dd,J=8.6,2.4Hz,1H),5.66(s,1H),4.29(dd,J=8.1,3.4Hz,1H),4.05–3.93(m,1H),3.88(dt,J=13.4,6.8Hz,2H),3.33–3.17(m,2H),2.84(dq,J=13.7,6.8Hz,1H),2.24(d,J=14.6Hz,1H),1.98(td,J=13.5,3.6Hz,1H),1.22(d,J=6.8Hz,6H),1.11(d,J=6.7Hz,6H),1.02(d,J=6.7Hz,6H);HRMS(ESI)calcd forC26H36N3O3S2[M+H]+502.2193,found502.2197.
实施例23
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺
(2)将2-溴-4-硝基苯甲醛(10g,43.48mmol)溶于100mL甲醇中,冰水浴下降温,待溶液温度降低,缓慢分批加入NaBH4(822mg,21.74mmol),搅拌反应20分钟,浓缩溶剂至甲醇量只有原来的1/10,再加入100mL水,搅拌半小时后抽滤,烘干,得白色固体10。将中间体10(5g,21.55mmol)溶于甲苯(80mL)中,加入溴乙酸叔丁酯(5.04g,25.86mmol),常温下搅拌,加入40%KOH水溶液(80mL)和0.1%的TBAB,室温条件下搅拌反应1h,加入乙酸乙酯,萃取三次,合并有机层,无水硫酸钠干燥、浓缩后得中间体11。将中间体11(5g,14.4mmol)溶于甲醇(80mL),加入甲醇钠(1.95g,36.11mmol),室温搅拌反应30min,向其中加入80mL水,室温下反应20min,浓缩溶剂,加入乙酸乙酯和水,合并水层,缓慢加入10%的盐酸使PH=2,加入乙酸乙酯萃取三次,合并有机层,无水硫酸钠干燥、浓缩得中间体12。将中间体12(3g,9.83mmol)用干燥的DCM(30mL)溶解,加入10mL草酰氯,加一滴DMF催化反应进行,室温下反应2h。此过程中,另称取N,O-二甲基羟胺盐酸盐(1.05g,10.82mmol),加入K2CO3(3.4g,24.58mmol),加入无水乙腈20mL,搅拌反应。待酰氯制备完全,浓缩DCM,将酰氯缓慢滴加入N,O-二甲基羟胺溶液中,室温下反应2h,浓缩溶剂,加入EA和H2O完全溶解,EA萃取三次,合并有机层,无水硫酸钠干燥,浓缩,PE:EA=1:1柱层析得中间体13。选取合适大小的三颈瓶,称量weinreb酰胺化合物13(2g,5.74mmol),氮气保护,用注射器注入重蒸的无水THF(20mL),搅拌溶解,-78℃下降温10分钟,严格无水无氧操作,用一次性注射器多次加入t-BuLi溶液(注意:量大需分批次加入叔丁基锂),-78℃下搅拌反应5分钟,此时溶液应成黄色澄清状,加入饱和氯化铵溶液(20mL)淬灭反应。加入乙酸乙酯及水适量,乙酸乙酯萃取3遍,合并有机层,无水硫酸钠干燥、浓缩溶剂,PE:EA=4:1柱层析得到中间体14。将甲基三苯基溴化膦(20.71mmol)加入到无水THF,0℃下,氩气保护,缓慢加入正丁基锂(18.64mmol),随后搅拌一个小时,加入溶于无水THF的中间体14(10.35mmol),升到室温,反应约一小时,之后用水淬灭反应,乙酸乙酯萃取,合并有机层,浓缩,用PE/EA=8:1柱层析得到中间体15。在0℃下将中间体15(7.85mmol)加入到碘(15.69mmol)和硫氰酸银(31.38mmol)的甲苯溶液中,室温搅拌过夜,抽滤,滤液用饱和亚硫酸钠洗,用乙酸乙酯萃取,合并有机层,随后将得到的有机层旋干,0℃下,将其溶于THF中并加入到NH3的乙醇溶液(2M,25mL),搅拌一个小时,随后在室温下搅拌2.5天。浓缩后用乙酸乙酯洗,抽滤,得中间体16,将其溶于THF中,加入三乙胺(28.27mmol)和二碳酸二叔丁酯(28.27mmol),室温反应2天,浓缩溶剂,直接用PE/EA=8:1柱层析得到中间体17。将中间体17(3.39mmol)加入到乙醇和饱和氯化铵溶液1:1的溶液(30mL)中,随后加入铁粉(6.79mmol),在87摄氏度下加热半个小时,趁热硅藻土过滤、乙酸乙酯萃取三次,合并有机层,浓缩溶剂后得到中间体18直接投下一步。将商业所得的吡啶-2-甲酸(0.67mmol)溶于20mL甲苯中,缓慢加入氯化亚砜(3.38mmol),并加入1滴DMF,在120度下反应1小时,冷却至室温后,浓缩溶剂得到吡啶-2甲酰氯备用。将吡啶-2甲酰氯(0.67mmol)溶于5mL的THF中,0℃下将该溶液加入至中间体18(0.59mmol)的THF(5mL)中,加入吡啶(30μL),随后升温到室温并且搅拌反应30分钟,随后用10mL饱和碳酸氢钠溶液淬灭,用乙酸乙酯萃取三次。浓缩溶剂得中间体19,将中间体19(0.59mmol)溶于10mL的无水二氯甲烷中,加入10mL三氟乙酸,室温反应2小时,随后用饱和碳酸氢钠溶液调PH=9。加入乙酸乙酯萃取三次,合并有机相,浓缩后用DCM:MeOH=20:1柱层析得到2-氨基噻唑类类终产物实施例23。白色固体,产率2.5%。1H NMR(500MHz,Chloroform-d)δ9.61(s,1H),8.73(dd,J=5.1,1.3Hz,1H),7.93(td,J=8.0,1.3Hz,1H),7.82(dd,J=8.0,1.0Hz,1H),7.63(ddd,J=7.9,5.0,1.0Hz,1H),7.42(dd,J=7.5,2.0Hz,1H),7.36(dt,J=7.4,1.0Hz,1H),7.13(d,J=2.0Hz,1H),6.25(s,2H),4.56–4.48(m,3H),4.46(d,J=0.9Hz,1H),3.68(d,J=6.6Hz,2H).13C NMR(125MHz,Chloroform-d)δ162.21,159.38,148.98,148.55,142.20,136.89,128.80,127.04,124.33,123.59,120.48,118.76,117.20,80.32,70.02,67.26,38.64.
实施例24
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺
同实施例23的操作,黄色固体,产率2.2%。1H NMR(300MHz,Chloroform-d)δ9.82(s,1H),8.50(d,J=1.1Hz,1H),7.77(d,J=8.0Hz,1H),7.69(dd,J=8.0,1.2Hz,1H),7.39(dd,J=7.5,2.0Hz,1H),7.33–7.26(m,2H),6.25(s,2H),4.56–4.48(m,3H),4.46(d,J=0.9Hz,1H),3.68(d,J=6.6Hz,2H).13C NMR(75MHz,Chloroform-d)δ162.28,160.58,148.31,141.37,138.58,130.25,128.80,128.53,128.27,125.78,120.02,118.88,117.76,117.62,78.89,70.31,67.26,38.64.
实施例25
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺
同实施例23的操作,白色固体,产率2.5%。1H NMR(300MHz,Chloroform-d)δ9.76(s,1H),8.52(d,J=1.2Hz,1H),7.93(d,J=8.0Hz,1H),7.43(dd,J=7.5,2.0Hz,1H),7.36(dt,J=7.6,1.1Hz,1H),7.28(d,J=1.9Hz,1H),7.05(dd,J=8.0,1.3Hz,1H),6.25(s,2H),4.56–4.48(m,3H),4.46(d,J=0.9Hz,1H),3.68(d,J=6.6Hz,2H).13C NMR(75MHz,Chloroform-d)δ162.21,160.58,156.58,147.44,141.37,138.88,128.53,128.36,125.31,121.95,118.76,117.94,117.03,78.89,69.85,67.26,38.64.
实施例26
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氯甲基吡啶酰胺
同实施例23的操作,白色固体,产率1.2%。1H NMR(300MHz,Chloroform-d)δ9.58(s,1H),8.47(d,J=1.2Hz,1H),7.88(d,J=8.0Hz,1H),7.44–7.34(m,3H),7.28(d,J=1.9Hz,1H),6.25(s,2H),4.53(d,J=11.0Hz,2H),4.48(dd,J=18.1,0.9Hz,2H),3.68(d,J=6.6Hz,2H).13C NMR(75MHz,Chloroform-d)δ162.21,160.58,147.44,141.95,141.37,130.50,129.47,128.80,128.53,125.31,120.28,118.88,117.94,78.89,69.85,67.26,38.64.
实施例27
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲氧基甲基吡啶酰胺
同实施例23的操作,白色固体,产率1.3%。1H NMR(300MHz,Chloroform-d)δ9.98(s,1H),8.11(d,J=1.3Hz,1H),7.88(d,J=8.0Hz,1H),7.39(dd,J=7.5,2.0Hz,1H),7.33–7.25(m,3H),6.25(s,2H),4.56–4.48(m,3H),4.46(d,J=0.9Hz,1H),3.83(s,2H),3.68(d,J=6.6Hz,2H).13C NMR(75MHz,Chloroform-d)δ162.21,160.58,152.96,143.97,141.37,134.12,128.80,128.53,125.78,123.97,122.73,118.88,117.62,78.89,70.31,67.26,56.63,38.64.
实施例28
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲基甲基吡啶酰胺
同实施例23的操作,白色固体,产率1.6%。1H NMR(300MHz,Chloroform-d)δ9.75(s,1H),8.21(d,J=1.2Hz,1H),7.88(d,J=8.0Hz,1H),7.47–7.32(m,2H),7.25(d,J=1.9Hz,1H),7.18(dd,J=8.0,1.3Hz,1H),6.25(s,2H),4.53(d,J=11.0Hz,2H),4.48(dd,J=18.1,0.9Hz,2H),3.68(d,J=6.6Hz,2H),2.31(s,3H).13CNMR(75MHz,Chloroform-d)δ162.17,160.58,148.31,143.97,141.37,134.90,128.96,128.36,127.83,125.31,124.26,118.76,117.94,78.89,69.85,67.26,37.36,17.80.
实施例29
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-3,5-二氯甲基吡啶酰胺
同实施例23的操作,白色固体,产率1.2%。1H NMR(300MHz,Chloroform-d)δ9.48(s,1H),8.47(d,J=1.3Hz,1H),7.54(d,J=1.1Hz,1H),7.38(dd,J=7.5,2.0Hz,1H),7.29(dt,J=7.5,0.9Hz,1H),7.25(d,J=2.0Hz,1H),6.25(s,2H),4.59(d,J=0.9Hz,1H),4.52–4.47(m,2H),4.42(s,1H),3.69(d,J=16.7Hz,2H).13CNMR(75MHz,Chloroform-d)δ162.32,159.91,141.14,140.72,135.77,131.93,131.16,129.48,129.24,129.01,125.78,118.88,117.94,81.33,70.31,67.26,38.64.
实施例30
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲氧基吡嗪-2-甲酰胺
同实施例23的操作,白色固体,产率1.5%。1H NMR(300MHz,Chloroform-d)δ9.76(s,1H),8.01(d,J=33.7Hz,2H),7.49–7.26(m,2H),7.25(d,J=2.0Hz,1H),6.25(s,2H),4.53(d,J=11.0Hz,2H),4.48(dd,J=18.1,0.9Hz,2H),3.92(s,3H),3.68(d,J=6.6Hz,2H).13C NMR(75MHz,Chloroform-d)δ163.49,162.17,160.58,142.00,137.14,135.91,133.30,128.80,128.53,125.78,118.88,117.62,78.89,70.31,67.26,54.26,38.64.
实施例31
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-3-氯-5-甲基甲基吡啶酰胺
同实施例23的操作,白色固体,产率1.5%。1H NMR(300MHz,Chloroform-d)δ9.58(s,1H),8.21(d,J=1.3Hz,1H),7.45–7.27(m,2H),7.25(d,J=1.5Hz,2H),6.25(s,2H),4.53(d,J=11.0Hz,2H),4.48(dd,J=18.1,0.9Hz,2H),3.68(d,J=6.6Hz,2H),2.40(s,3H).13C NMR(75MHz,Chloroform-d)δ162.32,160.58,143.97,142.00,137.34,135.77,131.54,129.76,128.66,128.53,125.95,118.88,117.62,78.89,70.31,67.26,38.64,17.77.
实施例32
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)苯酰胺
同实施例23的操作,白色固体,产率1.6%。1H NMR(300MHz,Chloroform-d)δ8.03–7.96(m,2H),7.59–7.52(m,1H),7.55–7.48(m,2H),7.42(dd,J=7.5,2.0Hz,1H),7.33(dt,J=7.5,1.1Hz,1H),7.25(d,J=2.0Hz,1H),6.25(s,2H),4.56–4.48(m,3H),4.46(d,J=0.9Hz,1H),3.68(d,J=6.6Hz,2H).13C NMR(75MHz,Chloroform-d)δ167.47,159.38,142.20,134.34,131.51,128.80,128.76,128.54,127.04,123.59,118.76,117.20,80.32,70.02,67.26,38.64.
实施例33
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-氟苯酰胺
同实施例23的操作,白色固体,产率1.3%。1H NMR(300MHz,Chloroform-d)δ8.05–7.99(m,2H),7.50(dd,J=7.5,2.0Hz,1H),7.44(d,J=2.0Hz,1H),7.33(dt,J=7.6,1.1Hz,1H),7.26–7.20(m,2H),6.25(s,2H),4.56–4.48(m,3H),4.46(d,J=0.9Hz,1H),3.68(d,J=6.6Hz,2H).13C NMR(75MHz,Chloroform-d)δ167.47,162.72,160.58,140.72,135.81,129.72,128.53,128.36,125.31,118.76,117.94,115.62,78.89,69.85,67.26,37.36.
实施例34
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)苯磺酰胺
同实施例23的操作,白色固体,产率1.5%。1H NMR(300MHz,Chloroform-d)δ7.84–7.73(m,2H),7.62–7.55(m,1H),7.52–7.43(m,3H),7.43–7.34(m,2H),6.25(s,2H),4.53(d,J=11.0Hz,2H),4.45(dd,J=11.5,0.9Hz,2H),3.68(d,J=6.6Hz,2H).13C NMR(75MHz,Chloroform-d)δ160.58,138.44,137.04,133.08,130.61,128.81,128.53,128.12,125.13,116.97,115.89,78.89,69.85,67.26,38.64.
实施例35
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氟苯磺酰胺
同实施例23的操作,白色固体,产率1.3%。1HNMR(300MHz,Chloroform-d)δ7.88–7.80(m,2H),7.49–7.39(m,2H),7.32(dt,J=7.4,1.0Hz,1H),7.27–7.16(m,2H),6.25(s,2H),4.53(d,J=11.0Hz,2H),4.45(dd,J=11.5,0.9Hz,2H),3.68(d,J=6.6Hz,2H).13C NMR(75MHz,Chloroform-d)δ162.87,160.58,138.44,136.18,130.61,128.53,128.13,125.54,117.60,116.88,116.71,78.89,70.31,67.26,38.64.
实施例36
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氯苯磺酰胺
同实施例23的操作,白色固体,产率1.2%。1HNMR(300MHz,Chloroform-d)δ9.93(s,1H),7.73–7.64(m,2H),7.58–7.50(m,2H),7.48–7.40(m,2H),7.30(dt,J=7.4,1.0Hz,1H),6.25(s,2H),4.51–4.47(m,3H),4.46(d,J=0.9Hz,1H),3.69(d,J=16.7Hz,2H).13CNMR(75MHz,Chloroform-d)δ159.91,138.44,136.89,136.21,129.76,129.19,127.97,125.54,117.94,117.60,81.33,70.31,67.26,38.64.
实施例37
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4溴苯磺酰胺
同实施例23的操作,白色固体,产率1.3%。1HNMR(300MHz,Chloroform-d)δ9.79(s,1H),7.72–7.56(m,4H),7.39(dd,J=7.5,2.0Hz,1H),7.36–7.23(m,2H),6.25(s,2H),4.54–4.41(m,4H),3.69(d,J=16.7Hz,2H).13C NMR(75MHz,Chloroform-d)δ159.91,138.31,136.31,130.76,129.76,129.18,127.65,125.54,123.97,118.07,117.94,81.33,70.31,67.26,37.36.
实施例38
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-(叔丁基)苯磺酰胺
同实施例23的操作,白色固体,产率1.6%。1H NMR(300MHz,Chloroform-d)δ9.66(s,1H),7.75–7.60(m,2H),7.48–7.36(m,4H),7.30(dt,J=7.4,1.0Hz,1H),6.25(s,2H),4.56–4.40(m,4H),3.69(d,J=16.7Hz,2H),1.30(s,9H).13C NMR(75MHz,Chloroform-d)δ159.35,148.64,138.98,136.82,129.76,128.53,127.82,126.41,125.21,118.15,117.94,84.63,69.98,67.05,38.64,34.79,31.19.
实施例39
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-(甲氧基)苯磺酰胺
同实施例23的操作,白色固体,产率1.8%。1H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.64–7.55(m,2H),7.43(d,J=1.9Hz,1H),7.42–7.21(m,2H),7.09–6.89(m,2H),6.25(s,2H),4.54–4.42(m,4H),3.86(s,3H),3.69(d,J=16.7Hz,2H).13C NMR(75MHz,Chloroform-d)δ159.61,159.35,138.98,136.16,129.76,128.79,127.83,125.36,117.60,116.88,114.76,84.63,69.98,67.05,55.19,38.64.
实施例40
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-甲基苯磺酰胺
同实施例23的操作,白色固体,产率1.5%。1H NMR(300MHz,Chloroform-d)δ9.92(s,1H),7.63–7.52(m,2H),7.46–7.38(m,2H),7.38–7.29(m,1H),7.17(dq,J=7.3,1.0Hz,2H),6.25(s,2H),4.53(d,J=11.0Hz,2H),4.45(dd,J=11.5,1.0Hz,2H),3.68(d,J=6.6Hz,2H),2.37(t,J=1.0Hz,3H).13C NMR(75MHz,Chloroform-d)δ160.58,142.86,138.44,136.71,130.61,128.83,128.57,128.53,125.54,117.60,116.88,78.89,70.31,67.26,38.64,21.26.
实施例41
N-(4-(N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)氨磺酰)苯基)乙酰胺
同实施例23的操作,白色固体,产率1.6%。1H NMR(300MHz,Chloroform-d)δ9.76(s,1H),7.96–7.64(m,4H),7.51–7.21(m,3H),6.25(s,2H),4.62–4.35(m,4H),3.69(d,J=16.7Hz,2H),2.07(s,3H).13C NMR(75MHz,Chloroform-d)δ169.05,159.35,141.01,138.98,136.38,129.76,128.62,128.53,125.21,119.78,118.15,117.94,84.63,69.98,67.05,38.64,24.27.
实施例42
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)萘-1-磺酰胺
同实施例23的操作,白色固体,产率1.8%。1H NMR(300MHz,Chloroform-d)δ10.00(s,1H),8.05(d,J=8.8Hz,2H),7.99(s,1H),7.85(s,1H),7.63(s,1H),7.51(s,1H),7.45(s,1H),7.43–7.35(m,2H),7.30(dt,J=7.4,1.0Hz,1H),6.25(s,2H),4.71–4.34(m,4H),3.69(d,J=16.7Hz,2H).13C NMR(75MHz,Chloroform-d)δ159.91,138.98,134.25,132.91,129.76,129.30,128.93,128.56,128.53,127.94,127.82,127.37,126.97,126.59,125.21,118.85,117.94,84.63,69.98,67.05,38.64.
实施例43
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)萘-2-磺酰胺
同实施例23的操作,白色固体,产率1.2%。1H NMR(300MHz,Chloroform-d)δ9.71(s,1H),8.39(s,1H),8.19–7.94(m,3H),7.85(s,1H),7.63(d,J=4.3Hz,2H),7.53–7.20(m,3H),6.25(s,2H),4.68–4.30(m,4H),3.69(d,J=16.7Hz,2H).13CNMR(75MHz,Chloroform-d)δ159.91,138.98,135.83,133.51,132.91,129.76,129.32,129.30,128.53,127.82,127.37,126.97,125.21,122.37,118.85,117.94,84.63,69.98,67.05,38.64.
实施例44
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-2,4,6-三异丙基苯磺酰胺
同实施例23的操作,白色固体,产率1.6%。1HNMR(300MHz,Chloroform-d)δ7.43(d,J=1.9Hz,1H),7.29(dt,J=7.5,1.1Hz,1H),7.24(dd,J=7.5,1.8Hz,1H),7.21(t,J=0.8Hz,2H),6.25(s,2H),4.64(d,J=0.9Hz,1H),4.54–4.43(m,3H),3.89(s,2H),3.69(d,J=16.7Hz,2H),2.91(t,J=1.0Hz,1H),1.26(d,J=24.9Hz,6H),1.19(d,J=24.9Hz,12H).13C NMR(75MHz,Chloroform-d)δ159.91,146.63,143.72,138.16,137.67,129.76,127.83,125.36,123.80,118.57,117.94,84.63,72.79,70.21,36.71,34.39,28.81,24.54,24.12.
实施例45
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺
操作同实施例23,其中将溴乙酸叔丁酯替换为2-溴丙酸叔丁酯。产物为白色固体,产率1.5%。
1H NMR(300MHz,Chloroform-d)δ9.75(s,1H),8.72(dd,J=5.0,1.2Hz,1H),7.93(td,J=8.0,1.3Hz,1H),7.82(dd,J=8.0,1.1Hz,1H),7.64(ddd,J=8.0,5.1,1.1Hz,1H),7.46–7.31(m,2H),7.25(d,J=1.9Hz,1H),6.25(s,2H),4.67(d,J=0.9Hz,1H),4.57–4.42(m,2H),3.62(d,J=26.2Hz,2H),1.34(s,3H).13C NMR(75MHz,Chloroform-d)δ162.17,157.86,148.98,148.55,141.37,136.81,130.97,125.99,125.31,125.26,120.48,118.76,114.71,91.57,75.02,68.23,41.63,22.60.
实施例46
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺
操作同实施例45,白色固体,产率1.4%。1H NMR(300MHz,Chloroform-d)δ9.79(s,1H),8.52(d,J=1.3Hz,1H),7.92(d,J=7.9Hz,1H),7.37(dd,J=7.4,1.9Hz,1H),7.36–7.23(m,2H),7.06(dd,J=8.0,1.2Hz,1H),6.25(s,2H),4.67(d,J=1.1Hz,1H),4.60(s,1H),4.54(d,J=1.1Hz,1H),3.62(d,J=26.2Hz,2H),1.34(s,3H).13C NMR(75MHz,Chloroform-d))δ162.21,157.86,156.58,145.44,141.37,137.76,131.05,126.53,125.78,121.95,118.88,117.03,114.71,97.36,75.02,67.84,41.63,22.60.
实施例47
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氯甲基吡啶酰胺
操作同实施例45,白色固体,产率1.1%。1H NMR(300MHz,Chloroform-d)δ9.82(s,1H),8.47(d,J=1.2Hz,1H),7.87(d,J=8.0Hz,1H),7.47–7.32(m,2H),7.30(dt,J=7.4,1.0Hz,1H),7.25(d,J=1.9Hz,1H),6.36(d,J=42.5Hz,2H),4.67(d,J=1.1Hz,1H),4.63–4.47(m,2H),3.62(d,J=26.2Hz,2H),1.35(s,3H).13C NMR(75MHz,Chloroform-d)δ162.21,158.53,147.44,144.65,141.37,131.05,130.50,129.33,127.49,125.95,120.05,118.88,114.71,97.36,75.02,68.23,41.63,22.55.
实施例48
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲氧基甲基吡啶酰胺
操作同实施例45,白色固体,产率1.3%。1H NMR(300MHz,Chloroform-d)δ8.11(d,J=1.3Hz,1H),7.88(d,J=8.0Hz,1H),7.36–7.21(m,4H),6.36(d,J=42.5Hz,2H),4.67(d,J=1.0Hz,1H),4.59(s,1H),4.54(d,J=1.0Hz,1H),3.82(s,3H),3.62(d,J=26.2Hz,2H),1.33(s,3H).13C NMR(75MHz,Chloroform-d)δ162.21,158.53,153.98,143.97,140.83,134.36,130.97,127.49,125.78,124.72,123.08,118.88,114.35,99.86,74.84,68.23,56.67,41.63,22.55.
实施例49
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲基甲基吡啶酰胺
操作同实施例45,白色固体,产率1.6%。1H NMR(300MHz,Chloroform-d)δ9.58(s,1H),8.21(d,J=1.2Hz,1H),7.88(d,J=8.0Hz,1H),7.39(dd,J=7.5,2.0Hz,1H),7.31(dt,J=7.5,1.1Hz,1H),7.25(d,J=2.0Hz,1H),7.18(dd,J=8.0,1.3Hz,1H),6.25(s,2H),4.67(d,J=1.1Hz,1H),4.60(s,1H),4.54(d,J=1.1Hz,1H),3.62(d,J=26.2Hz,2H),2.31(s,3H),1.34(s,3H).13C NMR(75MHz,Chloroform-d)δ162.28,157.86,145.44,143.97,141.37,134.79,131.05,128.96,125.99,125.78,124.26,118.88,114.71,97.36,75.02,67.84,41.63,22.60,17.77.
实施例50
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)苯磺酰胺
操作同实施例45,白色固体,产率1.5%。1H NMR(300MHz,Chloroform-d)δ9.92(s,1H),7.84–7.74(m,2H),7.64–7.52(m,1H),7.49–7.41(m,2H),7.38(dd,J=7.5,2.0Hz,1H),7.35–7.28(m,2H),6.36(d,J=42.5Hz,2H),4.67(d,J=1.1Hz,1H),4.59(s,1H),4.54(d,J=1.1Hz,1H),3.62(d,J=26.2Hz,2H),1.35(s,3H).13CNMR(75MHz,Chloroform-d)δ158.53,138.44,136.96,133.74,131.05,128.81,128.61,128.50,125.54,117.60,114.24,97.36,75.02,68.23,41.63,22.55.
实施例51
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氟苯磺酰胺
操作同实施例45,白色固体,产率1.3%。1H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.92–7.80(m,2H),7.38(dd,J=7.5,2.0Hz,1H),7.34(d,J=2.0Hz,1H),7.30(dt,J=7.5,1.0Hz,1H),7.26–7.16(m,2H),6.36(d,J=42.5Hz,2H),4.67(d,J=1.1Hz,1H),4.60(s,1H),4.54(d,J=1.1Hz,1H),3.62(d,J=26.2Hz,2H),1.33(s,3H).13C NMR(75MHz,Chloroform-d)δ163.31,157.86,138.98,136.32,130.97,128.50,128.13,125.36,117.60,116.54,114.24,99.86,74.84,68.23,41.63,22.55.
实施例52
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氯苯磺酰胺
操作同实施例45,白色固体,产率1.2%。1H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.74–7.64(m,2H),7.63–7.49(m,2H),7.43–7.24(m,3H),6.36(d,J=42.5Hz,2H),4.76–4.43(m,3H),3.61(d,J=37.2Hz,2H),1.33(s,3H).13C NMR(75MHz,Chloroform-d)δ158.53,138.98,136.57,136.19,130.97,129.19,128.50,127.97,125.36,117.94,114.45,99.86,74.58,68.23,41.63,22.55.
实施例53
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-(甲氧基)苯磺酰胺
操作同实施例45,白色固体,产率1.8%。1H NMR(300MHz,Chloroform-d)δ9.94(s,1H),7.67–7.51(m,2H),7.43–7.21(m,3H),7.08–6.83(m,2H),6.36(d,J=42.5Hz,2H),4.72–4.46(m,3H),3.85(s,3H),3.61(d,J=37.2Hz,2H),1.33(s,3H).13C NMR(75MHz,Chloroform-d)δ159.61,158.53,138.98,136.21,130.97,128.79,128.50,125.13,117.94,117.24,114.45,99.86,74.35,68.23,55.20,41.63,22.55.
实施例54
(R)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺
通过制备手性柱将实施例1拆分得到,产率100%。
实施例55
(S)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺
通过制备手性柱将实施例1拆分得到,产率100%。
实施例56
(R)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺
通过制备手性柱将实施例2拆分得到,产率100%。
实施例57
(S)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺
通过制备手性柱将实施例2拆分得到,产率100%。
实施例58
(R)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺
通过制备手性柱将实施例3拆分得到,产率100%。
实施例59
(S)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺
通过制备手性柱将实施例3拆分得到,产率100%。
实施例60
片剂
取上述配方,用常规方法制备成片剂。
以下是本发明部分化合物的药理实验结果:
部分化合物对BACE-1的体外酶活抑制实验
1.实验方法
化合物对BACE-1抑制活性的测定方法:
实验原理:将BACE-1置于最适的酶反应条件和反应体系中,将底物加到反应模板上,通过酶与底物的反应,用比色法来检测酶的活性。
实验材料:所有化合物样品均用二甲基亚砜(DMSO)配成10-2mol/L溶液,再用PH=4.5的醋酸钠缓冲溶液逐步稀释至所需浓度;BACE-1为人源酶。
测试方法:BACE-1抑制活性测试方法为FRET法。将测试化合物和阳性对照MK-8931用DMSO溶解,并用BACE-1缓冲液梯度稀释至所需浓度,并保证DMSO含量低于1%。往全黑的384孔板中依次加入:10μL BACE-1酶,10μLBACE-1底物和10μL不同浓度的化合物。室温孵育15min,随后在激发波长545nM,发射波长585nM波长下测定其荧光强度变化。抑制率的计算公式为:[1-(实验组荧光强度变化/空白组荧光强度变化)]×100%。测试IC50的方法为选取目标化合物的八个浓度测定BACE-1酶的抑制率(0.001–10μM),以摩尔浓度负对数与抑制率作线性回归,酶抑制率达到50%时的浓度即为该化合物的IC50值。
每个实验重复三次,实验结果表达为平均值±SEM。
1.实验结果:
表1化合物的IC50值
结果分析:我们利用FRET试验方法测定了实施例化合物对BACE-1的抑制活性,以默沙东公司开发的临床药物MK-8931作为阳性对照,从表1可以看出,所有化合物的BACE-1抑制活性均在纳摩尔级别,且优于阳性对照。
部分化合物对SH-SY5Y神经细胞的毒性
1.实验方法:
(1)SH-SY5Y细胞传代培养:弃去细胞培养瓶中的旧培养液,加入4mL D-Hanks轻轻摇晃,冲洗旧培养液中的FBS血清,结束后弃去洗涤液,加入2mL0.25%胰酶消化细胞1min,加4mL新鲜培养液终止消化,混匀后转移至20mL离心管中,在1000rpm下,离心10分钟,弃上清液,加入5mL新培养液以1:3比例进行传代培养;
(2)SH-SY5Y细胞种板:将10μL细胞悬液种于计数板中,倒置荧光显微镜计数为2×105个/mL接种SH-SY5Y细胞100μL/孔于96孔培养板中,于含5%CO2的37℃恒温培养箱内培养;
(3)换液:SH-SY5Y细胞培养24h后,将96孔培养板中的培养液换成MEM/F12培养液;
(4)化合物的配置与稀释:将待测化合物用DMSO溶解,用新鲜的MEM/F12培养液逐步稀释配置成所需测试浓度;
(5)给药:给药组每孔加入10μL相应浓度的待测化合物,对照组加入相同体积的溶剂,孵育2h后,各孔均加入10μL的MEM/F12培养液,并于含5%CO2的37℃恒温培养箱内培养;
(6)加MTT:SH-SY5Y细胞培养24h后,各孔加入10μL MTT,并于含5%CO2的37℃恒温培养箱内培养;
(7)测MTT:SH-SY5Y细胞继续培养3h后,弃去旧培养液,每孔加入100μLDMSO,振摇溶解20min,用FC酶标仪在490nM处测定每孔的吸光度值,计算给药后的细胞存活率和毒性。
2.实验结果:
表2部分化合物对SH-SY5Y的毒性
结果分析:从表2可以看出,Control组的SH-SY5Y细胞活力为100%,测试的化合物在浓度为1μM、10μM、50μM时细胞存活率均达到90%以上,该结果表明,这部分实施例对SH-SY5Y细胞无明显毒性。
部分化合物降低大鼠脑内Aβ40的水平:
1.实验方法:
雄性CD大鼠(体重约200g)分组群居,并在研究使用之前使它们适应动物饲养所5-7天。用20%羟基丙基-β-环糊精溶解化合物,口服给药10mg/kg。三小时后用过量的二氧化碳使大鼠安乐死,取出脑,用干冰冷冻。将所有的脑组织在-78℃下储存,随后用ELISA测定Aβ40水平。
2.实验结果:
表3口服10mg/kg化合物后3小时的大鼠皮层Aβ40变化
结果分析:从表3可以看出,这些化合物均能很好的降低大鼠脑内的Aβ40,且优于阳性药MK-8931。
尽管参照上面列出的具体实施方案描述了本发明,但许多备选方案、修饰及其它变化对本领域技术人员是显而易见的。所有这种备选方案、修饰和变体都要落在本发明的精神和范围之内。
Claims (10)
2.根据权利要求1所述的2-氨基噻唑类衍生物、异构体或其可药用的盐,其特征在于:Z为H或甲基或乙基。
3.根据权利要求1所述的2-氨基噻唑类衍生物、异构体或其可药用的盐,其特征在于:R2为取代或未取代的-N(H)-C(O)-R5,或取代或未取代的-N(H)-S(O2)-R6,R5或R6独立的选自苯、吡啶、吡嗪、噻吩、吡唑、噻唑、嘧啶、萘、呋喃、吡咯、茚、喹啉或吲哚;所述取代基选自H、卤素、-CN,-NO2、-OH,-COOH、-NH2、C1~C4烷基、C1~C4烷氧基、C1~C4卤代烷基、-N(H)-C(O)-C1~C4烷基或C1~C4烷氨基;优选的,所述的取代基选自H、卤素、-CH3、-CN、-NO2、-CF3、-OCH3、乙基、正丙基、异丙基、叔丁基或-N(H)-C(O)-CH3。
4.根据权利要求1所述的2-氨基噻唑类衍生物、异构体或其可药用的盐,其特征在于:Z为H或甲基;X为O或C;Y为O或C;R1,R3和R4为H;R2为取代或未取代的-N(H)-C(O)-R5,或取代或未取代的-N(H)-S(O2)-R6,R5或R6独立的选自苯、吡啶、吡嗪或萘,所述取代基选自:H、卤素、-CH3、-CN、,-NO2、-CF3、-OCH3、乙基、正丙基、异丙基、叔丁基或-N(H)-C(O)-CH3。
5.根据权利要求1所述的2-氨基噻唑类衍生物、异构体或其可药用的盐,其特征在于:Z为H或甲基;X为O或C;Y为O或C;R1,R3和R4为H;R2为取代或未取代的-N(H)-C(O)-R5或取代或未取代的-N(H)-S(O2)-R6,R5或R6独立的选自苯、吡啶、吡嗪或萘,所述取代基选自:H、卤素、-CH3、-CN、-OCH3、异丙基、叔丁基或-N(H)-C(O)-CH3。
6.如下所述化合物的任一种,或其异构体或可药用的盐:
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氯甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-甲氧基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-甲基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-3,5-二氯甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-甲氧基吡嗪-2-甲酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-3-氯-5-甲基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)苯酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-氟苯酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4氟苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4氯苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4溴苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-(叔丁基)苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-(甲氧基)苯磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-4-甲基苯磺酰胺;
N-(4-(N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)氨磺酰)苯基)乙酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)萘-1-磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)萘-2-磺酰胺;
N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-2,4,6-三异丙基苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氯甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲氧基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-3,5-二氯甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲氧基吡嗪-2-甲酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-3-氯-5-甲基甲基吡啶酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)苯酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-氟苯酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氟苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氯苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4溴苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-(叔丁基)苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-(甲氧基)苯磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-甲基苯磺酰胺;
N-(4-(N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)氨磺酰)苯基)乙酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)萘-1-磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)萘-2-磺酰胺;
N-(2'-氨基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-2,4,6-三异丙基苯磺酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-氯甲基吡啶酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲氧基甲基吡啶酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-5-甲基甲基吡啶酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)苯磺酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氟苯磺酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4氯苯磺酰胺;
N-(2'-氨基-3-甲基-5'H-螺[异色烷-4,4'-噻唑]-6-基)-4-(甲氧基)苯磺酰胺;
(R)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺;
(S)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)甲基吡啶酰胺;
(R)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺;
(S)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氰基甲基吡啶酰胺;
(R)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺;
(S)-N-(2'-氨基-5'H-螺[色烷-4,4'-噻唑]-6-基)-5-氟甲基吡啶酰胺。
8.一种药物组合物,包含权利要求1所述化合物、异构体或其可药用的盐,以及药学上可接受的载体。
9.权利要求1~6任一项所述的化合物、异构体或其可药用的盐,或者权利要求8所述药物组合物在制备BACE-1抑制剂药物中的应用。
10.权利要求1~6任一项所述的化合物、异构体或其可药用的盐,或者权利要求8所述药物组合物在制备抗痴呆症状药物中的应用;优选的,所述的痴呆症状包括阿尔茨海默病、额-颞叶痴呆、克-雅病、路易体痴呆、帕金森病或亨廷顿病。
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