CN118126023A - 吡唑酰胺类化合物及其制备方法与应用 - Google Patents
吡唑酰胺类化合物及其制备方法与应用 Download PDFInfo
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- CN118126023A CN118126023A CN202410180506.9A CN202410180506A CN118126023A CN 118126023 A CN118126023 A CN 118126023A CN 202410180506 A CN202410180506 A CN 202410180506A CN 118126023 A CN118126023 A CN 118126023A
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- Prior art keywords
- pyrazole
- hydroxy
- carboxamide
- phenyl
- compound
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- 230000002526 effect on cardiovascular system Effects 0.000 claims abstract description 4
- 150000001875 compounds Chemical class 0.000 claims description 125
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 75
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Abstract
本发明公开了一种吡唑酰胺类化合物及其制备方法与应用,涉及药物化学技术领域,所述吡唑酰胺类化合物包括如式(Ⅰ)所示的化合物或其药学上可接受的盐、异构体或者前药。本发明所涉及的吡唑酰胺类化合物能够有效的清除自由基起到抗氧化的作用,并且具有神经元保护作用。本发明提供的吡唑酰胺类化合物可在制备预防或治疗由自由基过量引发的脑卒中、心脑血管疾病、神经退行性疾病及其并发症的药品中的应用。
Description
技术领域
本发明涉及药物化学技术领域,具体涉及一种吡唑酰胺类化合物及其制备方法与应用。
背景技术
正常细胞有氧代谢会产生含氧和氮自由基,包括超氧阴离子自由基、一氧化氮自由基等。虽然氧气对好氧生物的生存至关重要,但它也会导致自由基的产生,尤其是生化过程中的活性氧(ROS),这是致命的。这些ROS是自由基,会引发一系列生化反应,破坏细胞成分,导致细胞坏死,并最终损害细胞成分,导致细胞坏死,最终导致各种病理状况,包括肝脏和血管疾病、炎症性疾病、类风湿性关节炎、氧化应激、癌症和衰老。大脑是体内氧糖符合最高、代谢最活跃的器官之一。因此,脑组织如神经元细胞和胶质细胞更易产生自由基,进而导致神经元损伤,进一步可能会引发神经退行性疾病。
脑血管栓塞引起脑组织缺血缺氧,进而引发脑组织缺血损伤级联反应。缺血后再灌注会诱导一系列神经炎症级联反应的发生,进一步加重脑组织死亡和血脑屏障损伤等。血流复灌导致氧气含量急剧升高,释放大量具有细胞毒性的活性氧自由基,诱导部分神经细胞死亡。在炎症介质和大量ROS富集等多重作用下,胶质细胞向促炎型转化导致炎症进一步恶化。因此,神经炎症的发生对于缺血性脑损伤具有时空特异性,且与胶质细胞激活、极化密切相关。
吡唑酮类化合物在异阴离子的形态存在时,可将结构中的一个电子提供给自由基,进而达到自由基清除的目的。其中,吡唑酮类化合物依达拉奉(Edaravone)是一种自由基清除剂,于2001年4月在日本首次上市,临床主要用于治疗缺血性脑卒中。因其分子结构中含有亲脂性基团,容易通过血脑屏障到达脑部组织,对脑缺血所致神经细胞损伤具有良好保护作用。依达拉奉通过清除氧自由基抑制脂质过氧化反应,调控炎症因子和抑制细胞凋亡等以达到对神经细胞的保护。研究表明除了治疗急性脑缺血外,依达拉奉对神经系统其他有氧化应激参与的疾病均有治疗作用。然而,依达拉奉产生的神经保护作用基本上依赖自由基清除作用,因此开发一种通过多途径发挥神经包含作用的药物十分必要。
发明内容
针对现有技术中依达拉奉产生的神经保护作用途径比较单一的问题,本发明的目的是提供一种吡唑酰胺类化合物及其制备方法与应用,可以多途径发挥挥神经包含作用的药物。
本发明的目的之一是提出了一种吡唑酰胺类化合物,所述化合物包含如式(Ⅰ)所示的化合物或其药学上可接受的盐、异构体或者前药:
其中,Ar1选自芳香环由不同数量和位置取代的烷基、烷氧基、卤素取代基团,优选的Ar1为:
X选自NH(CH2)n基团,其中n选自0~3的整数;
Ar2选自
所述的吡唑酰胺类化合物选自以下任一种化合物或其药学上可接受的盐、溶剂化物:
5-羟基-N-(6-甲基吡啶-3-基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(羟甲基)苯基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(3-(羟甲基)苯基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
N-(1,3-二氢异苯并呋喃-5-基)-5-羟基-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-((6-甲基吡啶-3-基)甲基)-1-苯基-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(羟甲基)苯基)-1-苯基-1H-吡唑-3-甲酰胺;
N-(2-(苯并[d][1,3]二氧杂环戊烯-4-基)乙基)-5-羟基-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
N-(3,5-二氯苄基)-5-羟基-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-1-(吡啶-2-基)-N-(吡啶-4-基甲基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(2-羟乙基)苯基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(6-(羟甲基)吡啶-3-基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(4-(羟甲基)苯基)-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(6-(羟甲基)吡啶-3-基)-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
N-(2-(苯并[d][1,3]二氧杂环戊烯-5-基)乙基)-1-(5-氟吡啶-2-基)-5-羟基-1H-吡唑-3-甲酰胺;
N-(苯并[d][1,3]二氧杂环戊烯-5-基)-1-(5-氟吡啶-2-基)-5-羟基-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(2-甲基吡啶-4-基)-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(3-(羟甲基)苯基)-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(3-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
1-(5-氯吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
1-(5-氯吡啶-2-基)-5-羟基-N-(3-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(2-羟乙基)苯基)-1-(嘧啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(2-羟乙基)苯基)-1-(3,5,6-三甲基吡嗪-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(2-羟乙基)苯基)-1-(5-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(3-(2-羟乙基)苯基)-1-(5-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(2-羟基-1-苯乙基)-1-(5-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺;
1-(5-氟嘧啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
1-(3,5-二氯吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(2-羟乙基)苯基)-1-(3-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺;
1-(3-氯吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
1-(4,6-二甲基嘧啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺。
本发明的目的之二是提出了一种吡唑酰胺类化合物制备方法,包括以下步骤:
(1)将化合物1溶于甲醇中制得摩尔浓度为0.85~1.5mol/L的溶液,在冰浴条件下滴加溶有0.9~1.5eq的丁炔二酸二甲酯的甲醇溶液,随后再向反应液中加入1.0~2.0eq甲醇钠,加热回流反应得到化合物2;
(2)将化合物2溶于体积比为4:3的甲醇和四氢呋喃的混合溶液中制得摩尔浓度为0.3~0.6mol/L的溶液,冰浴下滴加1.5~2.5eq的2mol/L的氢氧化钠水溶液后,反应制得化合物3;
(3)将化合物3溶于N,N-二甲基甲酰胺中制得摩尔浓度为0.05~0.1mol/L的溶液,再向反应液中加入2.0~3.5eq的N,N-二异丙基乙胺和1.0~1.5eq的缩合剂O-(7-氮杂苯并三唑-1-基)-N,N,N′,N′-四甲基脲六氟磷酸酯,最后向反应液中加入相应的1.0~1.5eq胺类化合物,制备吡唑酰胺类化合物4(I);
其中,所述胺类化合物包括6-甲基吡啶-3-胺、(4-氨基苯基)甲醇、(3-氨基苯基)甲醇、1,3-二氢异苯并呋喃-5-胺、3,4-亚甲二氧基苯乙胺、3,5-二氯苄胺、4-甲氨基吡啶、2-(4-氨基苯基)乙醇、2-(羟基)-5-氨基吡啶、对氨基苯甲醇、3,4-亚甲二氧苯乙胺、3,4-亚甲二氧苯胺、2-甲基-4-氨基吡啶、3-氨基苯甲醇、2-(3-氨基苯基)乙醇或DL-苯甘氨醇;
所述吡唑酰胺类化合物4(I)的合成路线如下:
本发明的目的之三是提出了一种包含所述的吡唑酰胺类化合物的药物组合物,由所述吡唑酰胺类化合物添加一种或多种药学上可接受的盐、异构体、前药、多晶型物或溶剂化合物。
进一步地,所述的药物组合物,所述制剂的剂型为胶囊剂、丸剂、片剂、颗粒剂或注射剂。
本发明的目的之四是提出了所述的吡唑酰胺类化合物的应用,包括在自由基清除中的应用、在神经元细胞保护中的应用以及在制备预防或治疗或辅助治疗由自由基过量引发的脑卒中、心脑血管疾病、神经退行性疾病及其并发症的药品中的应用。
有益效果:
与现有的技术相比,本发明具有如下显著的特点:本发明提出的吡唑酰胺类化合物能够有效清除自由基、能够改善由过氧化物导致的神经元细胞损伤。因此,该类化合物在制备预防或治疗由自由基过量引发的脑卒中、心脑血管疾病、神经退行性疾病中具有重要作用。
本发明的其它特征和优点将在随后的说明书中阐述,并且,部分地从说明书中变得显而易见,或者通过实施本发明而了解。本发明的目的和其他优点可通过在说明书、权利要求书以及附图中所指出的结构来实现和获得。
附图说明
图1为目标化合物的ORAC抗氧化能力表征图;
图2A和2B为目标化合物与不同金属的螯合能力表征图;
图3为目标化合物对由叔丁基过氧化氢(tBHP)诱导的人神经母细胞瘤细胞(SH-SY5Y)损伤的保护作用表征图;
图4为氧葡萄糖剥夺/再氧合(OGD/R)诱导的小胶质细胞(BV2)损伤的保护作用表征图;
图5为目标化合物对由tBHP诱导的活性氧(ROS)水平的影响结果。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚,下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地说明,显然,所描述的实施例是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
本发明实施例中所用溶剂均为市售化学纯或分析纯。通过核磁共振(NMR)确定化合物的结构。NMR的测定是使用Bruker AVANCE-300/500核磁共振仪,测定的溶剂是CDCl3或DMSO-d6,内标为TMS。
本发明提出的一种吡唑酰胺类化合物的制备方法包括以下步骤:
(1)将化合物1溶于甲醇中制得摩尔浓度为0.85~1.5mol/L的溶液,在冰浴条件下滴加溶有0.9~1.5eq的丁炔二酸二甲酯的甲醇溶液,随后再向反应液中加入1.0~2.0eq甲醇钠,加热回流反应得到化合物2;
(2)将化合物2溶于体积比为4:3的甲醇和四氢呋喃的混合溶液中制得摩尔浓度为0.3~0.6mol/L的溶液,冰浴下滴加1.5~2.5eq的2mol/L的氢氧化钠水溶液后,反应制得化合物3;
(3)将化合物3溶于N,N-二甲基甲酰胺中制得摩尔浓度为0.05~0.1mol/L的溶液,再向反应液中加入2.0~3.5eq的N,N-二异丙基乙胺和1.0~1.5eq的缩合剂O-(7-氮杂苯并三唑-1-基)-N,N,N′,N′-四甲基脲六氟磷酸酯,最后向反应液中加入相应的1.0~1.5eq胺类化合物,制备吡唑酰胺类化合物4(I)。
其中,所述胺类化合物包括6-甲基吡啶-3-胺、(4-氨基苯基)甲醇、(3-氨基苯基)甲醇、1,3-二氢异苯并呋喃-5-胺、3,4-亚甲二氧基苯乙胺、3,5-二氯苄胺、4-甲氨基吡啶、2-(4-氨基苯基)乙醇、2-(羟基)-5-氨基吡啶、对氨基苯甲醇、3,4-亚甲二氧苯乙胺、3,4-亚甲二氧苯胺、2-甲基-4-氨基吡啶、3-氨基苯甲醇、2-(3-氨基苯基)乙醇或DL-苯甘氨醇;
本发明提出的吡唑酰胺类化合物的合成路线图如下:
其中,Ar1选自芳香环由不同数量和位置取代的烷基、烷氧基、卤素取代基团,优选的Ar1为:
X选自NH(CH2)n基团,其中n选自0~3的整数;
Ar2选自 中任意一种。
以下结合具体实施例对上述吡唑酰胺类化合物的制备方法进行示例性说明。
实施例1:制备化合物5-羟基-1-苯基-1H-吡唑-3-甲酸甲酯(2a)
苯肼(1a,12g,110mmol)溶于甲醇(120mL),在冰浴条件下滴加溶有丁炔二酸二甲酯(15.2mL,124mmol)的甲醇(80mL)溶液,低温反应3小时后,向反应液中加入甲醇钠(9.2g,172mmol),70℃回流1小时。反应结束后,反应液浓缩除去甲醇,再加入80mL水,冰浴条件下滴加2mol/L HCl直至反应液pH=4,析出黄色沉淀,抽滤,水洗干燥滤饼,得黄色粉末2a,产率为60%。
实施例2:制备化合物5-羟基-1-苯基吡唑-3-羧酸(3a)
5-羟基-1-苯基-1H-吡唑-3-甲酸甲酯(2a,17.4g,80mmol)溶于甲醇(120mL)和四氢呋喃(90mL)混合液中,冰浴下滴加2mol/L NaOH(90mL,180mmol)后,反应液在40℃下回流5小时。反应结束后加2倍当量的水,反应液浓缩除去甲醇,冰浴条件下滴加2mol/L HCl至反应液pH=4,析出大量淡黄色固体,抽滤,水洗干燥滤饼,得淡黄色固体3a,产率为65%。
实施例3:制备化合物5-羟基-1-吡啶-2-基吡唑-3-羧酸(3b)
中间体3b是将苯肼(1a)替换为2-肼吡啶(1b)后与丁炔二酸二甲酯反应,再水解生成。反应过程与3a相似,产物为黄色粉末3b,产率为55%。1H NMR(300MHz,DMSO-d6)δ8.53(ddd,J=5.0,1.9,0.9Hz,1H),8.09(ddd,J=8.3,7.5,1.9Hz,1H),7.81(dt,J=8.3,1.0Hz,1H),7.46(ddd,J=7.5,5.0,1.1Hz,1H),5.99(s,1H)。
实施例4:制备化合物1-(5-氯吡啶-2-基)-5-羟基-1H-吡唑-3-羧酸(3c)
中间体3c是将苯肼(1a)替换为2-肼基-5-氟吡啶(1c)后与丁炔二酸二甲酯反应,再水解生成。反应过程与3a相似,产物为黄色固体3c,产率为58%。熔点:229.3℃-232.7℃。1HNMR(600MHz,DMSO-d6)δ8.56(d,J=3.0Hz,1H),7.99(td,J=8.6,3.1Hz,1H),7.80(dd,J=9.0,3.9Hz,1H),5.95(s,1H)。
实施例5:制备化合物1-(5-氯吡啶-2-基)-5-羟基-1H-吡唑-3-羧酸(3d)
中间体3d是将苯肼(1a)替换为2-肼基-5-氯吡啶(1d)后与丁炔二酸二甲酯反应,再水解生成。反应过程与3a相似,产物为深黄色固体3d,产率为70%。熔点:216.2℃-220.5℃。1H NMR(600MHz,DMSO-d6)δ12.14(s,1H),8.60(d,J=2.5Hz,1H),8.16(dd,J=8.7,2.6Hz,1H),7.80(d,J=8.7Hz,1H),5.98(s,1H)。
实施例6:制备化合物5-羟基-1-(嘧啶-2-基)-1H-吡唑-3-羧酸(3e)
中间体3e是将苯肼(1a)替换为2-肼基嘧啶(1e)后与丁炔二酸二甲酯反应,再水解生成。反应过程与3a相似,产物为橙色片状固体3e,产率为63%。熔点:162.4℃-172.9℃。1HNMR(600MHz,DMSO-d6)δ12.94(s,1H),12.02(s,1H),8.95(d,J=4.9Hz,2H),7.60(t,J=4.9Hz,1H),5.98(s,1H)。
实施例7:制备化合物5-羟基-1-(3,5,6-三甲基吡嗪-2-基)-1H-吡唑-3-羧酸(3f)
中间体3f是将苯肼(1a)替换为2-肼基-3,5,6-三甲基吡嗪(1f)后与丁炔二酸二甲酯反应,再水解生成。反应过程与3a相似,产物为黄色粉末3f,产率为55%。
实施例8:制备化合物5-羟基-1-(5-(三氟甲基)吡啶-2-基)-1H-吡唑-3-羧酸(3g)
中间体3g是将苯肼(1a)替换为2-肼基-5-三氟甲基吡啶(1g)后与丁炔二酸二甲酯反应,再水解生成。反应过程与3a相似,产物为橙红色粉末3g,产率为65%。熔点:137.4℃-144.5℃。1H NMR(600MHz,DMSO-d6)δ13.05(s,1H),12.31(s,1H),8.95(dt,J=2.8,1.0Hz,1H),8.44(dd,J=8.8,2.5Hz,1H),8.02(d,J=8.6Hz,1H),6.01(s,1H)。
实施例9:1-(3-氯吡啶-2-基)-5-羟基-1H-吡唑-3-羧酸(3h)
中间体3h是将苯肼(1a)替换为2-肼基-3-氯吡啶(1h)后与丁炔二酸二甲酯反应,再水解生成。反应过程与3a相似,产物为灰白色粉末3h,产率为60%。
实施例10:5-羟基-1-(3-(三氟甲基)吡啶-2-基)-1H-吡唑-3-羧酸(3i)
中间体3i是将苯肼(1a)替换为2-肼基-3-三氟甲基吡啶(1i)后与丁炔二酸二甲酯反应,再水解生成。反应过程与3a相似,产物为紫红色粉末3i,产率为65%。
实施例11:制备化合物1-(5-氟嘧啶-2-基)-5-羟基-1H-吡唑-3-羧酸(3j)
中间体3j是将苯肼(1a)替换为5-氟-2-肼基嘧啶1j后,与丁炔二酸二甲酯反应,再水解生成。反应过程与3a相似,产物为黄色固体3j,产率为70%。
实施例12:制备化合物1-(3,5-二氯吡啶-2-基)-5-羟基-1H-吡唑-3-羧酸(3k)
中间体3k是将苯肼(1a)替换为3,5-二氯-2-肼基吡啶(1k)后与丁炔二酸二甲酯反应,再水解生成。反应过程与3a相似,产物为土黄色固体3k,产率为73%。
实施例13:制备化合物1-(4,6-二甲基嘧啶-2-基)-5-羟基-1H-吡唑-3-羧酸(3l)
中间体3l是将苯肼(1a)替换为2-肼基-4,6-二甲基嘧啶(1l)后与丁炔二酸二甲酯反应,再水解生成。反应过程与3a相似,产物为白色固体3l,产率为80%。
实施例14:制备化合物5-羟基-N-(6-甲基吡啶-3-基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺(4-1)
将化合物3b(205mg,1mmol)溶于DMF(15mL)中,加入DIPEA(704μL,4mmol),冰浴下加入HATU(448mg,1.08mmol),加入化合物6-甲基吡啶-3-胺(108mg,1mmol),N2保护下,室温反应24h。并加入水(30mL)抽滤,滤饼经硅胶柱色谱得土黄色粉末4-1(190mg),产率64%。熔点:169.3℃-172.4℃。1H NMR(300MHz,DMSO-d6)δ10.51(s,1H),9.00(d,J=2.5Hz,1H),8.58(dd,J=5.0,1.7Hz,1H),8.35(dd,J=8.6,2.5Hz,1H),8.15(td,J=7.9,1.9Hz,1H),7.95(d,J=8.2Hz,1H),7.53(s,1H),7.50(d,J=3.6Hz,1H),6.11(s,1H),2.55(s,3H);13C NMR(75MHz,DMSO-d6)δ160.85,155.82,151.99,151.26,147.99,146.51,140.41,137.92,134.48,131.85,125.22,123.21,116.11,88.47,21.97;HRMS(m/z):calcd for C15H14N5O2[M+H]+,295.1190;found,295.1187。
实施例15:制备化合物5-羟基-N-(4-(羟甲基)苯基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺(4-2)
将化合物3b(205mg,1mmol)溶于DMF(15mL)中,加入DIPEA(528μL,3mmol),冰浴下加入HATU(414mg,1mmol),加入化合物(4-氨基苯基)甲醇(123mg,1mmol),N2保护下,室温反应24h。并加入水(30mL)抽滤,滤饼经硅胶柱色谱得淡黄色粉末4-2(200mg),产率64%。熔点:149.4℃-154.7℃。1H NMR(300MHz,DMSO-d6)δ10.02(s,1H),8.56-8.52(m,1H),8.17-8.09(m,1H),7.99(d,J=8.2Hz,1H),7.76(d,J=8.5Hz,2H),7.51-7.44(m,1H),7.29(d,J=8.5Hz,2H),6.08(s,1H),5.15(s,1H),4.47(s,2H).13C NMR(75MHz,DMSO-d6)δ160.19,155.94,152.39,147.68,147.38,140.53,138.32,137.56,127.31,122.85,120.48,115.51,88.34,63.11。
实施例16:制备化合物5-羟基-N-(3-(羟甲基)苯基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺(4-3)
将化合物3b(205mg,1mmol)溶于DMF(15mL)中,加入DIPEA(528μL,3mmol),冰浴下加入HATU(1.45g,3.5mmol),加入化合物(3-氨基苯基)甲醇(123mg,1mmol),N2保护下,室温反应24h。并加入水(30mL)抽滤,滤饼经硅胶柱色谱得灰棕色固体4-3(180mg),产率58%。熔点:160.8℃-165.8℃。1H NMR(300MHz,DMSO-d6)δ10.02(s,1H),8.58-8.52(m,1H),8.17-8.09(m,1H),8.01(d,J=8.2Hz,1H),7.79(s,1H),7.69(d,J=8.0Hz,1H),7.52-7.44(m,1H),7.30(t,J=7.8Hz,1H),7.06(d,J=7.6Hz,1H),6.09(s,1H),5.24(s,1H),4.50(s,2H).13C NMR(75MHz,DMSO-d6)δ160.21,155.93,152.46,147.62,147.34,143.59,140.56,138.82,128.78,122.83,122.27,119.08,118.83,115.40,88.37,63.39。
实施例17:制备化合物N-(1,3-二氢异苯并呋喃-5-基)-5-羟基-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺(4-4)
将化合物3b(205mg,1mmol)溶于DMF(15mL)中,加入DIPEA(440μL,2.5mmol),冰浴下加入HATU(448mg,1.08mmol),加入化合物1,3-二氢异苯并呋喃-5-胺(135mg,1mmol),N2保护下,室温反应24h。并加入水(30mL)抽滤,滤饼经硅胶柱色谱得白色粉末4-4(150mg),产率46%。熔点:188.8℃-190.2℃。1H NMR(300MHz,DMSO-d6)δ10.10(s,1H),8.55(ddd,J=4.9,1.8,0.8Hz,1H),8.13(ddd,J=8.2,7.5,1.8Hz,1H),7.98(d,J=8.3Hz,1H),7.82(s,1H),7.66(dd,J=8.2,1.7Hz,1H),7.48(ddd,J=7.4,5.0,1.0Hz,1H),7.28(d,J=8.2Hz,1H),6.08(s,1H),5.01(s,2H),4.99(s,2H).13CNMR(75MHz,DMSO-d6)δ160.27,155.89,152.37,147.69,147.30,140.52,139.98,138.23,134.55,122.88,121.60,119.99,115.54,113.40,88.37,73.07,72.86。
实施例18:制备化合物5-羟基-N-((6-甲基吡啶-3-基)甲基)-1-苯基-1H-吡唑-3-甲酰胺(4-5)
将化合物3a(204mg,1mmol)溶于DMF(15mL)中,加入DIPEA(528μL,3mmol),冰浴下加入HATU(448mg,1.08mmol),加入化合物6-甲基吡啶-3-胺(108mg,1mmol),N2保护下,室温反应24h。并加入水(30mL)抽滤,滤饼经硅胶柱色谱得淡黄色粉末4-5(180mg),产率61%。熔点:171.1℃-195.6℃。1H NMR(300MHz,DMSO-d6)δ10.26(s,1H),8.90(d,J=2.3Hz,1H),8.19(dd,J=8.5,2.5Hz,1H),7.85(m,J=8.5,1.0Hz,2H),7.57-7.50(m,2H),7.40(m,J=8.0,1.4Hz,1H),7.35(d,J=8.6Hz,1H),6.04(s,1H),2.48(s,3H).13C NMR(75MHz,DMSO-d6)δ161.04,154.38,151.89,145.69,139.52,138.58,134.07,130.29,129.46,127.43,124.27,122.75,88.22,22.76。
实施例19:制备化合物5-羟基-N-(4-(羟甲基)苯基)-1-苯基-1H-吡唑-3-甲酰胺(4-6)
将化合物3a(204mg,1mmol)溶于DMF(15mL)中,加入DIPEA(528μL,3mmol),冰浴下加入HATU(448mg,1.08mmol),加入化合物(4-氨基苯基)甲醇(123mg,1mmol),N2保护下,室温反应24h。并加入水(30mL)抽滤,滤饼经硅胶柱色谱得黄色粉末4-6(195mg),产率63%。熔点:174.5℃-182.6℃。1H NMR(300MHz,DMSO-d6)δ9.93(s,1H),7.88(d,J=7.9Hz,2H),7.77(d,J=8.1Hz,2H),7.54(t,J=7.8Hz,2H),7.40(d,J=7.2Hz,1H),7.31(s,1H),7.28(s,1H),6.04(s,1H),5.16(s,1H),4.48(s,2H).13C NMR(75MHz,DMSO-d6)δ160.56,154.29,146.32,138.71,138.13,137.70,129.44,127.29,127.24,122.57,120.43,88.11,63.12;HRMS(m/z):calcd for C17H16N3O3[M+H]+,310.1186;found,310.1185。
实施例20:制备化合物N-(2-(苯并[d][1,3]二氧杂环戊烯-4-基)乙基)-5-羟基-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺(4-7)
将化合物3b(410mg,2mmol)溶于DMF(7mL)中,加入DIPEA(696μL,4mmol),冰浴下加入HATU(912mg,2.4mmol),加入化合物3,4-亚甲二氧基苯乙胺(375μL,2.6mmol),N2保护下,室温反应24h。并加入水(30mL)抽滤,滤饼经硅胶柱色谱得淡黄色固体4-7(150mg),产率21%。熔点:79.5-87.8℃。1H NMR(600MHz,CDCl3)δ8.32(d,J=5.1Hz,1H),7.93(d,J=7.2Hz,1H),7.90(d,J=8.0Hz,1H),7.27-7.25(m,1H),7.03(d,J=6.5Hz,1H),6.77-6.74(m,2H),6.71-6.68(m,2H),6.07(d,J=4.7Hz,1H),5.94(s,1H),5.92(d,J=7.0Hz,1H),3.62(p,J=6.0,5.2Hz,2H),2.85(t,J=7.0Hz,2H).13C NMR(151MHz,CDCl3)δ161.63,157.20,154.08,147.79,147.57,146.17,145.38,140.20,132.72,121.76,121.04,112.40,109.23,108.35,100.89,88.33,40.61,35.67。
实施例21:制备化合物N-(3,5-二氯苄基)-5-羟基-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺(4-8)
将化合物3b(410mg,2mmol)溶于DMF(7mL)中,加入DIPEA(696μL,4mmol),冰浴下加入HATU(912mg,2.4mmol),加入化合物3,5-二氯苄胺(321μL,2.6mmol),N2保护下,室温反应24h。并加入水(30mL)抽滤,滤饼经硅胶柱色谱得白色固体4-8(210mg),产率30%。熔点:122.3℃-132.5℃。1H NMR(600MHz,CDCl3)δ12.73(s,1H),8.33(d,J=5.1Hz,1H),7.93(d,J=8.0Hz,2H),7.32(t,J=6.3Hz,1H),7.28-7.26(m,1H),7.25-7.23(m,2H),6.16(s,1H),4.58(d,J=6.3Hz,2H).13C NMR(151MHz,CDCl3)δ161.68,157.36,154.01,147.03,145.44,141.82,140.25,135.18,127.64,126.14,121.19,112.44,88.66,42.11。
实施例22:制备化合物5-羟基-1-(吡啶-2-基)-N-(吡啶-4-基甲基)-1H-吡唑-3-甲酰胺(4-9)
将化合物3b(410mg,2mmol)溶于DMF(7mL)中,加入DIPEA(696μL,4mmol),冰浴下加入HATU(912mg,2.4mmol),加入化合物4-甲氨基吡啶(263μL,2.6mmol),N2保护下,室温反应24h。并加入水(30mL)抽滤,滤饼经硅胶柱色谱得白色固体4-9(218mg),产率37%。熔点:99.5℃-101.7℃。1H NMR(600MHz,CDCl3)δ8.57(d,J=6.1Hz,2H),8.34(d,J=5.1Hz,1H),7.95-7.90(m,2H),7.42(t,J=6.3Hz,1H),7.30(d,J=6.0Hz,2H),7.28-7.26(m,1H),6.16(s,1H),4.66(d,J=6.3Hz,2H);13C NMR(151MHz,CDCl3)δ161.93,157.37,153.99,149.75(×2),147.81,146.99,145.49,140.26,122.42(×2),121.24,112.44,88.65,41.94;HRMS(m/z):calcd for C15H12N5O2[M-H]-,294.0996;found,294.1026。
实施例23:制备化合物5-羟基-N-(4-(2-羟乙基)苯基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺(4-10)
将化合物3b(410mg,2mmol)溶于DMF(5mL)中,加入DIPEA(528μL,3mmol),冰浴下加入HATU(912mg,2mmol),加入化合物(2-(4-氨基苯基)乙醇(328.96mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得黄色固体4-10(170mg),产率27.5%。熔点:125.5℃-132.7℃。1H NMR(600MHz,DMSO-d6)δ9.96(s,1H),8.54(dd,J=5.1,1.6Hz,1H),8.12(td,J=7.9,1.8Hz,1H),7.98(d,J=8.2Hz,1H),7.70(d,J=8.2Hz,2H),7.47(dd,J=7.4,5.0Hz,1H),7.19(d,J=8.2Hz,2H),6.08(s,1H),4.65(s,1H),3.60(t,J=7.1Hz,2H),2.70(t,J=7.1Hz,2H).13CNMR(151MHz,DMSO-d6)δ160.12,155.93,152.36,147.71,147.40,140.48,136.86,135.35,129.43,122.85,120.68,115.58,88.33,62.70,38.97;HRMS(m/z):calcd for C17H17N4O3[M+H]+,325.1295;found,325.1291。
实施例24:制备化合物5-羟基-N-(6-(羟甲基)吡啶-3-基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺(4-11)
将化合物3b(410mg,2mmol)溶于DMF(7mL)中,加入DIPEA(528μL,3mmol),冰浴下加入HATU(836mg,2.2mmol),加入化合物2-(羟基)-5-氨基吡啶(272.88mg,2.2mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得黄色固体4-11(185mg),产率29%。熔点:169.3℃-174.7℃。1H NMR(600MHz,DMSO-d6)δ10.29(s,1H),8.89(d,J=2.4Hz,1H),8.56(d,J=5.7Hz,1H),8.21(dd,J=8.5,2.5Hz,1H),8.13(td,J=7.8,1.8Hz,1H),7.97(d,J=8.2Hz,1H),7.49(t,J=5.7Hz,1H),7.46(d,J=8.5Hz,1H),6.10(s,1H),4.55(s,2H).13C NMR(151MHz,DMSO-d6)δ160.67,157.22,155.85,152.22,147.82,146.91,141.35,140.46,134.04,128.56,123.01,120.51,115.79,88.41,64.45;HRMS(m/z):calcdfor C15H14N5O3[M+H]+,312.1091;found,312.1086。
实施例25:制备化合物1-(5-氟吡啶-2-基)-5-羟基-N-(4-(羟甲基)苯基)-1H-吡唑-3-甲酰胺(4-12)
将化合物3c(223mg,1mmol)溶于DMF(5mL)中,加入DIPEA(216μL,1.2mmol),冰浴下加入HATU(760mg,3mmol),加入化合物对氨基苯甲醇(184.60mg,1.2mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得灰白色固体4-12(200mg),产率60%。熔点:167.5℃-170.4℃。1H NMR(600MHz,DMSO-d6)δ9.95(s,1H),8.57(d,J=3.0Hz,1H),8.02(td,J=8.5,3.0Hz,1H),7.97(dd,J=9.0,4.1Hz,1H),7.75(d,J=8.2Hz,2H),7.27(d,J=8.2Hz,2H),5.99(s,1H),5.13(s,1H),4.46(s,2H);13C NMR(151MHz,DMSO)δ160.39,158.23(d,J=252.5Hz),155.70,147.88,147.17,138.20,137.63,136.17(d,J=26.0Hz),127.27,126.90(d,J=20.5Hz),120.44,119.06(d,J=5.4Hz),87.98,63.11;HRMS(m/z):calcd for C16H14FN4O3[M+H]+,329.1044;found,329.1044。
实施例26:制备化合物1-(5-氟吡啶-2-基)-5-羟基-N-(6-(羟甲基)吡啶-3-基)-1H-吡唑-3-甲酰胺(4-13)
将化合物3c(446mg,2mmol)溶于DMF(7mL)中,加入DIPEA(418μL,2.4mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物2-(羟甲基)-5-氨基吡啶(297.73mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得黄色固体4-13(210mg),产率32%。熔点:190.1℃-200.3℃。1H NMR(600MHz,DMSO-d6)δ12.12(s,1H),10.29(s,1H),8.88(d,J=2.5Hz,1H),8.59(d,J=3.0Hz,1H),8.20(dd,J=8.5,2.5Hz,1H),8.05(td,J=8.6,3.0Hz,1H),7.93(dd,J=9.0,4.0Hz,1H),7.45(d,J=8.4Hz,1H),6.05(s,1H),5.38(s,1H),4.54(s,2H).13C NMR(151MHz,DMSO-d6)δ160.76,158.40(d,J=252.9Hz),157.13,155.08,147.64,146.71,141.35,136.34(d,J=26.1Hz),134.09,128.54,126.97(d,J=20.2Hz),120.46,119.40(d,J=5.4Hz),88.26,64.43;HRMS(m/z):calcd for C15H13FN5O3[M+H]+,330.0997;found,330.0995。
实施例27:制备化合物1-(5-氟吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺(4-14)
将化合物3c(446mg,2mmol)溶于DMF(7mL)中,加入DIPEA(418μL,2.4mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物2-(4-氨基苯基)乙醇(329mg,2.4mmol),N2保护下,室温反应24h。加入水(35mL)抽滤,滤饼经硅胶色谱柱黄色固体化合物4-14(310mg),产率45%。熔点:124.5℃-129.5℃。1H NMR(600MHz,DMSO-d6)δ9.93(s,1H),8.57(d,J=3.0Hz,1H),8.03(td,J=8.6,3.0Hz,1H),7.95(dd,J=9.0,4.0Hz,1H),7.69(d,J=8.2Hz,2H),7.18(d,J=8.2Hz,2H),6.01(s,1H),4.64(s,1H),3.59(t,J=7.1Hz,2H),2.69(t,J=7.1Hz,2H).13C NMR(151MHz,DMSO)δ160.24,158.28(d,J=252.7Hz),155.31,147.83(d,J=2.5Hz),147.19,136.91,136.19(d,J=26.2Hz),135.24,129.40(×2),126.97(d,J=20.2Hz),120.67(×2),119.12(d,J=5.1Hz),88.14,62.71,38.98。
实施例28:制备化合物N-(2-(苯并[d][1,3]二氧杂环戊烯-5-基)乙基)-1-(5-氟吡啶-2-基)-5-羟基-1H-吡唑-3-甲酰胺(4-15)
将化合物3c(446mg,2mmol)溶于DMF(7mL)中,加入DIPEA(418μL,2.4mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物3,4-亚甲二氧苯乙胺(396.17mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得灰白色固体4-15(410mg),产率55%。熔点:129.3℃-136.3℃。1H NMR(600MHz,DMSO-d6)δ11.91(s,1H),8.55(d,J=3.0Hz,1H),8.21(t,J=5.9Hz,1H),8.01(td,J=8.6,3.0Hz,1H),7.83(dd,J=9.3,3.9Hz,1H),6.82(d,J=8.1Hz,1H),6.81(s,1H),6.68(dd,J=7.9,1.6Hz,1H),5.96(s,2H),5.90(s,1H),3.45-3.39(m,2H),2.75(t,J=7.4Hz,2H).13C NMR(151MHz,DMSO-d6)δ161.57,158.18(d,J=252.3Hz),154.76,147.85,147.64,147.21,145.94,136.14(d,J=26.2Hz),133.69,126.99(d,J=20.2Hz),121.93,118.72(d,J=4.5Hz),109.44,108.58,101.10,87.82,40.66,35.30;HRMS(m/z):calcd for C18H16FN4O4[M+H]+,371.1150;found,371.1146。
实施例29:制备化合物N-(苯并[d][1,3]二氧杂环戊烯-5-基)-1-(5-氟吡啶-2-基)-5-羟基-1H-吡唑-3-甲酰胺(4-16)
将化合物3c(446mg,2mmol)溶于DMF(7mL)中,加入DIPEA(418μL,2.4mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物3,4-亚甲二氧苯胺(328.89mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得黄色粉末4-16(500mg),产率70%。熔点:165.6℃-169.4℃。1H NMR(600MHz,DMSO-d6)δ12.01(s,1H),9.95(s,1H),8.58(d,J=3.0Hz,1H),8.04(td,J=8.6,3.0Hz,1H),7.93(dd,J=9.1,3.9Hz,1H),7.47(d,J=2.0Hz,1H),7.26(dd,J=8.4,2.0Hz,1H),6.88(d,J=8.4Hz,1H),6.02(s,1H),6.00(s,2H).13C NMR(151MHz,DMSO-d6)δ160.12,158.32(d,J=252.8Hz),154.98,147.80,147.39,147.15,143.65,136.20(d,J=26.2Hz),133.34,127.02(d,J=20.3Hz),119.18(d,J=4.1Hz),113.73,108.35,102.85,101.43,88.18;HRMS(m/z):calcd for C16H12FN4O4[M+H]+,343.0837;found,343.0833。
实施例30:制备化合物1-(5-氟吡啶-2-基)-5-羟基-N-(2-甲基吡啶-4-基)-1H-吡唑-3-甲酰胺(4-17)
将化合物3c(446mg,2mmol)溶于DMF(7mL)中,加入DIPEA(522μL,3mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物2-甲基-4-氨基吡啶(259.36mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶色谱柱得黄色粉末化合物4-17(200mg),产率31%。1H NMR(600MHz,DMSO-d6)δ10.29(s,1H),8.57(d,J=2.9Hz,1H),8.32(d,J=5.6Hz,1H),8.02(td,J=8.6,2.9Hz,1H),7.96(dd,J=9.5,3.9Hz,1H),7.73(d,J=1.9Hz,1H),7.65(d,J=5.7Hz,1H),5.99(s,1H),2.43(s,3H)。
实施例31:制备化合物1-(5-氟吡啶-2-基)-5-羟基-N-(3-(羟甲基)苯基)-1H-吡唑-3-甲酰胺(4-18)
将化合物3c(446mg,2mmol)溶于DMF(7mL)中,加入DIPEA(522μL,3mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物3-氨基苯甲醇(295.36mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得褐红色固体4-18(200mg),产率30%。熔点:193.4℃-196.7℃。1H NMR(600MHz,DMSO-d6)δ9.98(s,1H),8.58(d,J=2.9Hz,1H),8.04(td,J=8.6,3.0Hz,1H),7.97(dd,J=9.0,4.0Hz,1H),7.79(t,J=1.8Hz,1H),7.67(dd,J=8.0,2.1Hz,1H),7.29(t,J=7.8Hz,1H),7.05(d,J=7.5Hz,1H),6.04(s,1H),5.22(s,1H),4.50(s,2H).13C NMR(151MHz,DMSO-d6)δ160.30,158.29(d,J=252.6Hz),155.08,147.89,147.11,143.54,138.84,136.14(d,J=26.2Hz),128.74,127.06(d,J=20.2Hz),122.20,119.07,119.02(d,J=4.7Hz),118.83,88.22,63.38;HRMS(m/z):calcd for C16H14FN4O3[M+H]+,329.1044;found,329.1040。
实施例32:制备化合物1-(5-氟吡啶-2-基)-5-羟基-N-(3-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺(4-19)
将化合物3c(446mg,2mmol)溶于DMF(7mL)中,加入DIPEA(522μL,3mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物2-(3-氨基苯基)乙醇(329.mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得黄色粉末4-19(150mg),产率22%。熔点:177.7℃-185.4℃。1H NMR(600MHz,DMSO-d6)δ12.04(s,1H),9.93(s,1H),8.58(d,J=3.0Hz,1H),8.04(td,J=8.6,3.0Hz,1H),7.96(dd,J=9.0,4.0Hz,1H),7.70-7.61(m,2H),7.31-7.15(m,1H),6.95(d,J=7.5Hz,1H),6.04(s,1H),4.68(s,1H),3.62(t,J=7.1Hz,2H),2.72(t,J=7.1Hz,2H).13C NMR(151MHz,DMSO)δ160.27,158.30(d,J=252.5Hz),155.06,147.89,147.13,140.38,138.85,136.15(d,J=26.4Hz),128.80,127.06(d,J=20.3Hz),124.73,121.18,119.06(d,J=5.2Hz),118.34,88.20,62.60,40.51;HRMS(m/z):calcd for C17H16FN4O3[M+H]+,343.1201;found,343.1197。
实施例33:制备化合物1-(5-氯吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺(4-20)
将化合物3d(478mg,2mmol)溶于DMF(7mL)中,加入DIPEA(522μL,3mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物2-(4-氨基苯基)乙醇(329.24mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得淡黄色粉末4-20(235mg),产率32.8%。熔点:168.8℃-171.6℃。1H NMR(600MHz,DMSO-d6)δ9.97(s,1H),8.61(d,J=2.5Hz,1H),8.20(dd,J=8.7,2.6Hz,1H),7.97(d,J=8.8Hz,1H),7.69(d,J=8.5Hz,2H),7.18(d,J=8.5Hz,2H),6.05(s,1H),4.64(s,1H),3.60(t,J=7.1Hz,2H),2.70(t,J=7.1Hz,2H).13CNMR(151MHz,DMSO-d6)δ160.14,155.74,150.19,147.58,146.79,139.51(×2),136.87,135.31,129.41(×2),120.69(×2),118.05,88.41,62.70,38.97;HRMS(m/z):calcd forC17H16ClN4O3[M+H]+,359.0905;found,359.0901。
实施例34:制备化合物1-(5-氯吡啶-2-基)-5-羟基-N-(3-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺(4-21)
将化合物3d(478mg,2mmol)溶于DMF(7mL)中,加入DIPEA(522μL,3mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物2-(3-氨基苯基)乙醇(329.24mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得黄色粉末4-21(239mg),产率33.4%。熔点:203.3℃-206.9℃。1H NMR(600MHz,DMSO-d6)δ12.12(s,1H),9.96(s,1H),8.62(d,J=2.6Hz,1H),8.21(dd,J=8.8,2.6Hz,1H),7.98(d,J=8.7Hz,1H),7.66(d,J=7.2Hz,1H),7.65(s,1H),7.30-7.19(m,1H),7.03-6.93(m,1H),6.06(s,1H),4.68(s,1H),3.62(t,J=7.1Hz,2H),2.72(t,J=7.1Hz,2H).13C NMR(151MHz,DMSO-d6)δ160.17,155.35,150.22,147.49,146.78,140.40,139.60,138.80,129.52,128.82,124.79,121.20,118.36,118.09,88.46,62.60,40.49。
实施例35:制备化合物5-羟基-N-(4-(2-羟乙基)苯基)-1-(嘧啶-2-基)-1H-吡唑-3-甲酰胺(4-22)
将化合物3e(412.32mg,2mmol)溶于DMF(7mL)中,加入DIPEA(522μL,3mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物2-(4-氨基苯基)乙醇(329.24mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶色谱柱得淡黄色粉末4-22(280mg),产率43%。熔点:241.5℃-252.1℃。1H NMR(600MHz,DMSO-d6)δ10.00(s,1H),8.97(d,J=4.9Hz,2H),7.70(d,J=8.5Hz,2H),7.61(t,J=4.9Hz,1H),7.18(d,J=8.5Hz,2H),6.07(s,1H),4.65(s,1H),3.59(t,J=7.2Hz,2H),2.69(t,J=7.1Hz,2H).13C NMR(151MHz,DMSO-d6)δ160.21,159.57(×2),156.46,156.11,148.24,136.85,135.33,129.40(×2),120.78(×2),120.67,88.39,62.72,38.98。
实施例36:制备化合物5-羟基-N-(4-(2-羟乙基)苯基)-1-(3,5,6-三甲基吡嗪-2-基)-1H-吡唑-3-甲酰胺(4-23)
将化合物3f(372mg,1.5mmol)溶于DMF(7mL)中,加入DIPEA(392μL,2.25mmol),冰浴下加入HATU(855mg,2.25mmol),加入化合物2-(4-氨基苯基)乙醇(246.69mg,1.8mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得黄色固体4-23(170mg),产率31%。熔点:192.5℃-199.3℃。1H NMR(600MHz,DMSO-d6)δ11.96(s,1H),9.89(s,1H),7.68(d,J=8.5Hz,2H),7.15(d,J=8.3Hz,2H),5.99(s,1H),4.63(s,1H),3.58(t,J=7.1Hz,2H),2.68(t,J=7.2Hz,2H),2.56(s,3H),2.50(s,3H),2.30(s,3H).13C NMR(151MHz,DMSO-d6)δ160.35,154.89,153.61,149.58,147.43,147.29,142.09,136.98,135.08,129.32(×2),120.71(×2),86.86,62.72,38.97,21.81,21.31,19.54。
实施例37:制备化合物5-羟基-N-(4-(2-羟乙基)苯基)-1-(5-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺(4-24)
将化合物3g(546mg,2mmol)溶于DMF(8mL)中,加入DIPEA(522μL,3mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物2-(4-氨基苯基)乙醇(329.24mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶色谱柱得黄色固体4-24(180mg),产率22%。熔点:172.5℃-179.4℃。1H NMR(600MHz,DMSO-d6)δ12.29(s,1H),10.05(s,1H),8.96(d,J=2.5Hz,1H),8.50(dd,J=8.8,2.3Hz,1H),8.19(d,J=8.6Hz,1H),7.70(d,J=8.2Hz,2H),7.20(d,J=8.2Hz,2H),6.09(s,1H),4.66(s,1H),3.60(t,J=7.1Hz,2H),2.70(t,J=7.1Hz,2H).13C NMR(151MHz,DMSO-d6)δ159.92,156.10,154.53,148.34,145.66(q,J=4.6Hz),137.48(q,J=4.0Hz),136.78,135.45,129.44(×2),123.98(q,J=272.0Hz),123.59(q,J=32.7Hz),120.76(×2),116.12,88.85,62.69,38.97。
实施例38:制备化合物5-羟基-N-(3-(2-羟乙基)苯基)-1-(5-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺(4-25)
将化合物3g(546mg,2mmol)溶于DMF(8mL)中,加入DIPEA(522μL,3mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物2-(3-氨基苯基)乙醇(329.24mg,2.4mmol),N2保护下,室温反应24h。并加入水(35mL)抽滤,滤饼经硅胶柱色谱得白色粉末4-25(220mg),产率28%。熔点:158.4℃-163.0℃。1H NMR(600MHz,DMSO-d6)δ12.32(s,1H),10.04(s,1H),9.05-8.86(m,1H),8.50(dd,J=8.8,2.4Hz,1H),8.21(d,J=8.6Hz,1H),7.75-7.58(m,2H),7.25(t,J=7.7Hz,1H),6.97(d,J=7.5Hz,1H),6.09(s,1H),4.69(s,1H),3.62(t,J=7.1Hz,2H),2.72(t,J=7.1Hz,2H).13C NMR(151MHz,DMSO-d6)δ159.99,156.11,154.58,148.27,145.65(q,J=4.3Hz),140.45,138.73,137.48(q,J=3.9Hz),128.84,124.89,123.99(q,J=272.0Hz),123.58(q,J=33.2Hz),121.27,118.43,116.09,88.83,62.59,39.66。
实施例39:制备化合物5-羟基-N-(2-羟基-1-苯乙基)-1-(5-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺(4-26)
将化合物3g(546mg,2mmol)溶于DMF(8mL)中,加入DIPEA(522μL,3mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物DL-苯甘氨醇(329.24mg,2.4mmol),N2保护下,室温反应24h。加入水(35mL)抽滤,滤饼经硅胶柱色谱得白色固体4-26(300mg),产率38.26%。熔点:126.4℃-131.5℃。1H NMR(600MHz,DMSO-d6)δ12.28(s,1H),8.95(d,J=2.3Hz,1H),8.55-8.40(m,2H),8.13(d,J=8.8Hz,1H),7.39(d,J=7.6Hz,2H),7.33(t,J=7.5Hz,2H),7.24(t,J=7.3Hz,1H),5.97(s,1H),5.04(q,J=7.4Hz,2H),3.75(dd,J=11.1,7.3Hz,1H),3.71(dd,J=11.3,5.3Hz,1H).13C NMR(151MHz,DMSO-d6)δ161.14,155.86,154.40,148.22,145.73(q,J=4.5Hz),141.55,137.43(q,J=3.9Hz),128.57(×2),127.43(×2),127.32,123.99(q,J=271.9Hz),123.51(q,J=32.8Hz),116.04,88.56,64.69,55.46。
实施例40:制备化合物1-(5-氟嘧啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺(4-27)
将化合物3j(360mg,1.5mmol)溶于DMF(7mL)中,加入DIPEA(392μL,2.25mmol),冰浴下加入HATU(855mg,2.25mmol),加入化合物2-(4-氨基苯基)乙醇(247mg,1.8mmol),N2保护下,室温反应24h。加入水(20mL)无固体析出,遂85℃减压浓缩除去DMF,加水(15mL)后用EA萃取3次,浓缩有机相后经硅胶柱色谱得淡黄色固体4-27(200mg),产率37%。熔点:70.5℃-82.5℃。1H NMR(600MHz,DMSO-d6)δ11.97(br s,1H),9.97(s,1H),9.07(s,2H),7.69(d,J=8.0Hz,2H),7.17(d,J=8.0Hz,2H),6.03(s,1H),4.63(br s,1H),3.59(t,J=7.3Hz,2H),2.70(d,J=7.4Hz,2H).13C NMR(151MHz,DMSO-d6)δ160.21,156.74(d,J=261.1Hz),155.64,152.30(d,J=3.5Hz),148.02,147.54(d,J=22.7Hz,2H),136.88,135.29,129.37(×2),120.78(×2),88.23,62.71,38.98。
实施例41:制备化合物1-(3,5-二氯吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺(4-28)
将化合物3k(409mg,1.5mmol)溶于DMF(5mL)中,加入DIPEA(392μL,2.25mmol),冰浴下加入HATU(855mg,2.25mmol),加入化合物2-(4-氨基苯基)乙醇(247mg,1.8mmol),N2保护下,室温反应24h。加入水(25mL)无固体析出,遂85℃减压浓缩除去DMF,加水(20mL)后用EA萃取3次,浓缩有机相后经硅胶柱色谱得白色固体4-28(200mg),产率34%。熔点:155.8℃-166.4℃。1H NMR(600MHz,DMSO-d6)δ11.98(br s,1H),10.00(s,1H),8.78(d,J=2.3Hz,1H),8.65(d,J=2.3Hz,1H),7.73(d,J=8.2Hz,2H),7.20(d,J=8.2Hz,2H),6.03(s,1H),4.67(br s,1H),3.63(t,J=7.1Hz,2H),2.73(t,J=7.1Hz,2H).13C NMR(151MHz,DMSO-d6)δ160.20,155.29,148.04,146.92,145.79,139.58,136.95,135.14,133.13,129.97,129.32(×2),120.75(×2),86.77,62.72,38.98。
实施例42:制备化合物5-羟基-N-(4-(2-羟乙基)苯基)-1-(3-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺(4-29)
将化合物3i(448mg,2mmol)溶于DMF(7mL)中,加入DIPEA(522μL,3mmol),冰浴下加入HATU(1.14g,3mmol),加入化合物2-(4-氨基苯基)乙醇(329mg,2.4mmol),N2保护下,室温反应24h。加入水(25mL)抽滤,滤饼经硅胶柱色谱得白色粉末4-26(150mg),产率22%。熔点:213.4℃-220.8℃。1H NMR(600MHz,DMSO-d6)δ9.78(s,1H),9.05-8.86(m,1H),8.49(d,J=8.0Hz,1H),7.93-7.77(m,1H),7.67(d,J=8.1Hz,2H),7.14(d,J=8.1Hz,2H),5.90(s,1H),4.75-4.44(m,1H),3.57(t,J=7.2Hz,2H),2.67(t,J=7.2Hz,2H).13C NMR(151MHz,DMSO-d6)δ160.42,156.57,153.43,147.42,147.14,138.01(q,J=4.6Hz),136.96,135.08,129.33(×2),126.18,123.55(q,J=32.8Hz),122.89(q,J=273.5Hz),120.64(×2),86.22,62.72,38.97。
实施例43:制备化合物1-(3,5-二氯吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺(4-30)
将化合物3h(411mg,1.5mmol)溶于DMF(7mL)中,加入DIPEA(392μL,2.25mmol),冰浴下加入HATU(855mg,2.25mmol),加入化合物2-(4-氨基苯基)乙醇(247mg,1.8mmol),N2保护下,室温反应24h。加入水(35mL)抽滤,滤饼经硅胶柱色谱得橙黄色固体4-30(120mg),产率20%。熔点:145.6℃-158.4℃。1H NMR(600MHz,DMSO-d6)δ12.13(s,1H),9.96(s,1H),8.67(dd,J=4.5,1.5Hz,1H),8.31(dd,J=8.1,1.6Hz,1H),7.79-7.62(m,3H),7.20(d,J=8.2Hz,2H),6.02(s,1H),4.68(s,1H),3.63(t,J=7.2Hz,2H),2.73(t,J=7.1Hz,2H).13CNMR(151MHz,DMSO-d6)δ160.35,148.16,147.59,147.19,140.13,136.99,135.08,130.47,129.47,129.32(×2),127.27,120.72(×2),86.60,62.72,38.97。
实施例44:制备化合物1-(4,6-二甲基嘧啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺(4-31)
将化合物3l(351mg,1.5mmol)溶于DMF(6mL)中,加入DIPEA(392μL,2.25mmol),冰浴下加入HATU(855mg,2.25mmol),加入化合物2-(4-氨基苯基)乙醇(247mg,1.8mmol),N2保护下,室温反应24h。加入水(20mL)抽滤,滤饼经硅胶柱色谱得白色固体4-31(220mg),产率41.5%。熔点:238.4℃-245.8℃。1H NMR(600MHz,DMSO-d6)δ12.27(s,1H),10.01(s,1H),7.69(d,J=8.4Hz,2H),7.36(s,1H),7.17(d,J=8.2Hz,2H),6.07(s,1H),4.64(s,1H),3.67-3.49(m,2H),2.69(t,J=7.1Hz,2H),2.53(s,6H).13C NMR(151MHz,DMSO-d6)δ169.25,160.25,156.31,156.21,148.22,136.86,135.35,129.38(×2),120.81(×2),119.26,88.23,62.70,40.38,40.25,40.11,39.97,39.83,39.69,39.55,38.98,23.78。
实施例45:DPPH自由基清除实验
DPPH测试液的配置:取DPPH固体1.35mg溶于34.3mL甲醇,配置为100μM的DPPH测试液。样品液的配置:样品用合适的溶剂溶解配制成30mM的溶液。具体加样如表1所示。在测试前,将30mM的待测化合物溶液用甲醇稀释为300μM,依达拉奉(Eda)作为阳性对照化合物。再用甲醇稀释为5个浓度(10,20,40,60,80μM)进行测试。A值测量(待测组):在96孔板中,用移液枪取40μL的待测化合物的甲醇溶液,再加入160μL的DPPH溶液。以上所有测试均进行两次平行操作。在室温下10h,测A519nm值。A0值测量(空白组):用移液枪取40μL的甲醇溶液,在加入160μL的DPPH溶液,测出的A值即为A0。实验结果(表2)表明:所有的目标化合物在4~16μM浓度下均对DPPH具有清除能力。其中,大部分目标化合物显示出比依达拉奉更优的DPPH清除能力。部分化合物在16μM浓度下对DPPH的清除能力达到70%以上(依达拉奉在同等浓度下的清除率为37.2%)。
表1.DPPH测试加样表
待测化合物溶液 | 甲醇 | DPPH测试液 | 总体积 |
30μL | 10μL | 160μL | 200μL |
25μL | 15μL | 160μL | 200μL |
20μL | 20μL | 160μL | 200μL |
10μL | 30μL | 160μL | 200μL |
5μL | 35μL | 160μL | 200μL |
表2.化合物对DPPH的清除能力(%)
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实施例46:ORAC荧光测试抗氧化能力
通过氧自由基吸收能力-荧光素(ORAC-FL)测定化合物的抗氧化活性。Trolox(Tro)作为阳性对照化合物。在96孔黑板中依次加入待测试化合物(20μL,20μM)与荧光素溶液(120μL,125nM)。将测试板置于37℃下孵育15分钟。随后向体系中加入偶氮二异丁脒盐酸盐(AAPH,60μL,40mM),最终体积为200μL,开始反应。加入AAPH后,立即在Tecan 96酶标仪(lex 485nm,lem 525nm,37℃)中每分钟记录一次荧光,持续120分钟。将反应混合物一式三份制备。在未使用抗氧化剂或不含AAPH和抗氧化剂的情况下测量空白对照组。使用为每种测定计算的标准曲线将ORAC值TE表示为Trolox当量(平均值±SD)。抗氧化测试的实验结果(表3和图1中未示出所有化合物)表明,所有化合物均能够清除由偶氮类化合物AAPH产生的过氧自由基,本发明实施例中的化合物相较于Trolox均展现出优异的抗氧化能力。其中化合物4-23展现出最优的抗氧化能力,其ORAC值达到2.43。
表3.化合物的抗氧化能力测试数据(TE)
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实施例47:金属螯合测试
以化合物4-24为例,本实施例采用紫外光谱法探究了目标化合物的金属螯合能力。测试所采用的金属盐有氯化铝、氯化亚铜、氯化铜、硫酸亚铁、氯化铁和氯化锌。将测试化合物(1.0mL,300μM)的甲醇溶液加入至含金属离子(1.0mL,300μM)的甲醇溶液中,室温静置2小时。然后将溶液转移至石英池中测量溶液在200-550nm处的吸光度。实验结果(图2A和图2B)表明,代表性化合物分别与Cu2+和Fe2+溶液孵育后产生吸光谱线红移,Cu2+表现更为明显。
实施例48:叔丁基过氧化氢诱导的神经元细胞损伤的保护作用评价
神经细胞的选择包括但不局限于SH-SY5Y、PC-12细胞。实验方法为:分别取对数生长期细胞一瓶,加入胰酶细胞消化液消化,使贴壁细胞脱落,配制成浓度为1×105个/mL的细胞悬液,96孔细胞培养板中每孔加入100μL细胞悬液(每孔1×104个细胞),板置于37℃,5% CO2培养箱中培养24小时。用DMEM培养液配制细胞损伤剂,如叔丁基过氧化氢(tBHP)等。再用含有细胞损伤剂的DMEM培养基配制待测试的化合物,并将含有待测试化合物的培养基与对应的细胞孵育36小时。随后弃去培养液,PBS缓冲盐溶液清洗1次,每孔加入100μL相应的含不同浓度待测化合物的DMEM(含损伤剂)培养液;同时设空白对照组(只加高糖DMEM培养液)。96孔板置于37℃,5% CO2培养箱中培养48小时,每孔加入15μL MTT溶液(5mg/mL),在培养箱继续培养4小时。弃去培养基,每孔加入150μL DMSO溶解,摇床震荡5min使结晶完全溶解。最后以多功能酶标仪于490nm波长处读取OD值,计算细胞的生长抑制率。结果表明:所有的化合物对由叔丁基过氧化氢诱导的神经细胞损伤具有保护作用,部分化合物的实验结果如图3所示,并且其中部分化合物的神经细胞保护作用优于依达拉奉。
实施例49:氧糖剥夺/再氧合(OGD/R)诱导的胶质细胞损伤的保护作用评价
胶质细胞的选择包括但不局限于BV2细胞。实验方法为:分别取对数生长期细胞一瓶,加入胰酶细胞消化液消化,使贴壁细胞脱落,配制成浓度为4×104个/mL的细胞悬液,96孔细胞培养板中每孔加入100μL细胞悬液(每孔1×104个细胞),板置于37℃,5%CO2培养箱中培养24小时。24小时后,将培养基改为不含葡萄糖和血清的DMEM培养基,并放入含有5%CO2和95%N2的三气体培养箱中培养2小时。然后用含有待测试化合物且含有10% FBS的完全培养基培养细胞,并将细胞置于含有5%CO2和20%O2的湿润气氛中培养48小时。最后向每个孔中加入20μL MTT溶液(5mg/mL)。培养后,丢弃每个孔的上清液,并向每个孔中加入150μL DMSO,在细胞振荡器上振荡10分钟。最后以多功能酶标仪于490nm波长处读取OD值,计算细胞的生长抑制率。部分化合物的实验结果如图4所示:代表性化合物对由OGD/R诱导的BV2细胞损伤具有保护作用,能够显著改善由缺氧缺葡萄糖导致的细胞损伤和皱缩(图4)。
实施例50:细胞内活性氧(ROS)水平的检测
选取对数生长期的SH-SY5Y细胞,调整细胞浓度至5.0×104个/mL,以0.4mL/孔Z种于24孔板,置恒温CO2培养箱中培养24h。先用待测试化合物作用1h,再用含对应待测试化合物的tBHP(175μM)继续作用24h。去除原培养基,每孔加入用无血清培养基配置(1:1000稀释)的浓度为10μM的DCFH-DA 0.1mL,37℃细胞培养箱内孵育20分钟。每隔4分钟混匀一下,使探针和细胞充分接触。去除细胞培养液,用不含血清的培养基洗涤细胞三次,以充分去除未进入细胞内的DCFH-DA。在荧光倒置显微镜下观察并记录绿色荧光,整个操作需在10分钟内结束,以防止荧光猝灭。绿色荧光的强度与细胞内ROS的含量成正比(图5)。实验结果表明化合物能够有效抑制细胞内ROS的生成。
尽管参照前述实施例对本发明进行了详细的说明,本领域的普通技术人员应当理解:其依然可以对前述各实施例所记载的技术方案进行修改,或者对其中部分技术特征进行等同替换;而这些修改或者替换,并不使相应技术方案的本质脱离本发明各实施例技术方案的精神和范围。
Claims (10)
1.一种吡唑酰胺类化合物,其特征在于,包含如式(Ⅰ)所示的化合物或其药学上可接受的盐、异构体或者前药:
其中,Ar1选自芳香环由不同数量和位置取代的烷基、烷氧基或卤素取代基团,具体Ar1为:
X选自-NH(CH2)n基团,其中n选自0~3的整数;
Ar2选自 中任意一种。
2.根据权利要求1所述的吡唑酰胺类化合物,其特征在于,选自以下任一种化合物或其药学上可接受的盐:
5-羟基-N-(6-甲基吡啶-3-基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(羟甲基)苯基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(3-(羟甲基)苯基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
N-(1,3-二氢异苯并呋喃-5-基)-5-羟基-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-((6-甲基吡啶-3-基)甲基)-1-苯基-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(羟甲基)苯基)-1-苯基-1H-吡唑-3-甲酰胺;
N-(2-(苯并[d][1,3]二氧杂环戊烯-4-基)乙基)-5-羟基-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
N-(3,5-二氯苄基)-5-羟基-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-1-(吡啶-2-基)-N-(吡啶-4-基甲基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(2-羟乙基)苯基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(6-(羟甲基)吡啶-3-基)-1-(吡啶-2-基)-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(4-(羟甲基)苯基)-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(6-(羟甲基)吡啶-3-基)-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
N-(2-(苯并[d][1,3]二氧杂环戊烯-5-基)乙基)-1-(5-氟吡啶-2-基)-5-羟基-1H-吡唑-3-甲酰胺;
N-(苯并[d][1,3]二氧杂环戊烯-5-基)-1-(5-氟吡啶-2-基)-5-羟基-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(2-甲基吡啶-4-基)-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(3-(羟甲基)苯基)-1H-吡唑-3-甲酰胺;
1-(5-氟吡啶-2-基)-5-羟基-N-(3-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
1-(5-氯吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
1-(5-氯吡啶-2-基)-5-羟基-N-(3-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(2-羟乙基)苯基)-1-(嘧啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(2-羟乙基)苯基)-1-(3,5,6-三甲基吡嗪-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(2-羟乙基)苯基)-1-(5-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(3-(2-羟乙基)苯基)-1-(5-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(2-羟基-1-苯乙基)-1-(5-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺;
1-(5-氟嘧啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
1-(3,5-二氯吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
5-羟基-N-(4-(2-羟乙基)苯基)-1-(3-(三氟甲基)吡啶-2-基)-1H-吡唑-3-甲酰胺;
1-(3-氯吡啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺;
1-(4,6-二甲基嘧啶-2-基)-5-羟基-N-(4-(2-羟乙基)苯基)-1H-吡唑-3-甲酰胺。
3.如权利要求1所述的吡唑酰胺类化合物,其特征在于,所述异构体为酮-烯醇互变异构形成的互变异构体。
4.一种如权利要求1-3任一项所述的吡唑酰胺类化合物的制备方法,其特征在于,包括以下步骤:
(1)将化合物1溶于甲醇中制得摩尔浓度为0.85~1.5mol/L的溶液,在冰浴条件下滴加溶有0.9~1.5eq的丁炔二酸二甲酯的甲醇溶液,随后再向反应液中加入1.0~2.0eq甲醇钠,加热回流反应得到化合物2;
(2)将化合物2溶于体积比为4:3的甲醇和四氢呋喃的混合溶液中制得摩尔浓度为0.3~0.6mol/L的溶液,冰浴下滴加1.5~2.5eq的2mol/L的氢氧化钠水溶液后,反应制得化合物3;
(3)将化合物3溶于N,N-二甲基甲酰胺中制得摩尔浓度为0.05~0.1mol/L的溶液,再向反应液中加入2.0~3.5eq的N,N-二异丙基乙胺和1.0~1.5eq的缩合剂O-(7-氮杂苯并三唑-1-基)-N,N,N′,N′-四甲基脲六氟磷酸酯,最后向反应液中加入相应的1.0~1.5eq胺类化合物,制备吡唑酰胺类化合物4(I);
其中,所述胺类化合物包括6-甲基吡啶-3-胺、(4-氨基苯基)甲醇、(3-氨基苯基)甲醇、1,3-二氢异苯并呋喃-5-胺、3,4-亚甲二氧基苯乙胺、3,5-二氯苄胺、4-甲氨基吡啶、2-(4-氨基苯基)乙醇、2-(羟基)-5-氨基吡啶、对氨基苯甲醇、3,4-亚甲二氧苯乙胺、3,4-亚甲二氧苯胺、2-甲基-4-氨基吡啶、3-氨基苯甲醇、2-(3-氨基苯基)乙醇或DL-苯甘氨醇;
所述吡唑酰胺类化合物4(I)的合成路线如下:
5.一种药物组合物,其特征在于,包含权利要求1-3任一项所述的吡唑酰胺类化合物。
6.一种药物制剂,其特征在于,由权利要求1-3任一项所述的吡唑酰胺类化合物添加一种或多种药学上可接受的辅料制成制剂。
7.根据权利要求6所述的药物制剂,其特征在于,所述制剂的剂型为胶囊剂、丸剂、片剂、颗粒剂或注射剂。
8.如权利要求1-3任一项所述的吡唑酰胺类化合物或按照权利要求4所述的制备方法制备的吡唑酰胺类化合物在自由基清除中的应用。
9.如权利要求1-3任一项所述的吡唑酰胺类化合物或按照权利要求4所述的制备方法制备的吡唑酰胺类化合物在神经元细胞保护中的应用。
10.如权利要求1-3任一项所述的吡唑酰胺类化合物或按照权利要求4所述的制备方法制备的吡唑酰胺类化合物在制备预防或治疗或辅助治疗由自由基过量引发的脑卒中、心脑血管疾病、神经退行性疾病及其并发症的药品中的应用。
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