CN117164591A - 吡咯并嘧啶类化合物,其制备方法及制药用途 - Google Patents
吡咯并嘧啶类化合物,其制备方法及制药用途 Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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Abstract
本发明涉及一类含吡咯并嘧啶类结构的化合物或其药学上可接受盐,其制备方法,含有该类化合物的药物组合物以及该类化合物的医药用途。本发明化合物具有特异性抑制BTK的作用,且对TMD‑8淋巴瘤细胞抑制作用较强,具有较高的生物利用度且在TMD‑8肿瘤模型上表现出了较强的抑制活性,能够用于制备治疗B细胞恶性肿瘤、自身免疫性疾病等与BTK功能相关的疾病的药物。
Description
技术领域
本发明涉及药物领域,具体涉及一类含吡咯并嘧啶类结构的化合物或其药学上可接受的盐,其制备方法,含有该类化合物的药物组合物以及该类化合物在制备治疗与布鲁顿式酪氨酸激酶(Bruton’s tyrosine kinase,BTK)功能相关的疾病的药物中的应用。
背景技术
酪氨酸蛋白激酶(Tyrosine protein kinase,TPK)是一类催化ATP上γ-磷酸转移到蛋白酪氨酸残基上的激酶,能催化多种底物蛋白质酪氨酸残基磷酸化,在细胞信号转导通路中占据了十分重要的作用,调节细胞生长、分化、死亡等一系列生理生化过程。酪氨酸蛋白激酶功能失调将引发生物体内一系列疾病。
BTK是酪氨酸蛋白激酶Tec家族的主要成员之一。BTK是B细胞抗原受体(B-cellreceptor,BCR)信号转导通路中的关键激酶,在BCR信号通路中,磷酸化的PI3K(phosphatidylinositol 3-kinase)将膜上的PIP2转化为第二信使PIP3。PIP3结合到BTK的PH结构域,BTK随后会被募集到细胞膜,随后Tyr-551残基被SYK和LYN激酶磷酸化。BTK接着在Tyr-223残基进行自磷酸化反应从而具备生理活性。活化的BTK激活许多下游通路,如下游的蛋白激酶C,核因子kappa-B(NF-κB),MAPK通路等,对B细胞多个细胞过程,如生长、发育、分化、凋亡产生重要影响。
BTK在B细胞相关恶性肿瘤,如急性B淋巴细胞白血病,非霍奇金性淋巴瘤,在自身免疫性疾病,如类风湿关节炎、系统性红斑狼疮等疾病中扮演着重要的角色。这使得BTK成为B细胞恶性肿瘤和自身免疫性疾病的潜在靶点。现有BTK抑制剂选择性差,生物利用度低,导致临床用药量较大,毒副作用较大。因此,亟需发现活性高,成药性好的新型BTK抑制剂。
发明内容
本发明提供了一类含吡咯并嘧啶类结构的化合物或其药学上可接受的盐,其制备方法,含有该类化合物的药物组合物以及该类化合物在制备治疗与BTK功能相关的疾病的药物中的应用。
为解决本发明的技术问题,本发明提供如下技术方案:
本发明技术方案的第一方面是提供如通式(I)所示的化合物或其药学上可接受的盐:
其中,R选自且R3选自丙烯酰基,2-丁炔酰基;
R1,R2取代基为一个或多个,选自H,C1-8直链或支链烷基,C3-8环烷基,C1-8烷氧基,C1-8烷基氨基,卤素,羟基,氨基,氰基,且所述的C1-8直链或支链烷基进一步被一个或多个卤素,羟基,氨基或氰基取代。
进一步地,根据权利要求1的化合物或其药学上可接受的盐,其特征在于R1,R2选自H,甲基,乙基,正丙基,异丙基,正丁基,异丁基,叔丁基,正戊基,异戊基,环丙基,环丁基,环戊基,甲氧基,乙氧基,丙氧基,甲基氨基,乙基氨基,F,Cl,Br,I,羟基,氨基,氰基,或者选自被一个或多个卤素取代的甲基,乙基,正丙基,异丙基,正丁基,异丁基,叔丁基,正戊基,异戊基。
进一步地,根据权利要求1的化合物或其药学上可接受的盐,其特征在于R1选自H,甲基,乙基,三氟甲基,F,Cl,Br,或I;R2选自H,异丙基,异丁基,叔丁基,异戊基,环丙基,环丁基、环戊基。
所述药学上可接受的盐选自有机酸盐或无机酸盐,包括但不限于盐酸盐,氢溴酸盐,硫酸盐,磷酸盐,醋酸盐,柠檬酸盐,苹果酸盐,富马酸盐,酒石酸盐或甲磺酸盐。
作为本发明的优选方案,通式(I)的化合物具有如下的结构:
本发明技术方案的另一方面是提供如通式(I)所示的化合物或其药学上可接受的盐的合成方法,其合成路线如下:
路线(1):
试剂与条件:(a)(2-(氯甲氧基)乙基)三甲基硅烷,碳酸钾,N,N-二甲基甲酰胺,室温;(b)N-Boc-1,2,5,6-四氢吡啶-4-硼酸频哪醇酯,1,1'-二(二苯膦基)二茂铁二氯化钯,碳酸钾,1,4-二氧六环,100℃;(c)R1,R2取代苯基硼酸频哪醇酯,1,1'-二(二苯膦基)二茂铁二氯化钯,碳酸钾,1,4-二氧六环,100℃;(d)盐酸的乙醇溶液,室温;丙烯酰氯,N,N-二异丙基乙胺,二氯甲烷,室温;(e)三氟乙酸,二氯甲烷,室温;四氢呋喃溶液,氢氧化钠溶液,室温;
路线(2):
试剂与条件:(a)(2-(氯甲氧基)乙基)三甲基硅烷,碳酸钾,N,N-二甲基甲酰胺,室温;(b)N-Boc-1,2,5,6-四氢吡啶-4-硼酸频哪醇酯,1,1'-二(二苯膦基)二茂铁二氯化钯,碳酸钾,1,4-二氧六环,100℃;(c)R1,R2取代苯基硼酸频哪醇酯,1,1'-二(二苯膦基)二茂铁二氯化钯,碳酸钾,1,4-二氧六环,100℃;(d)氢气,钯碳,乙酸乙酯溶液,甲醇溶液,室温(e)盐酸的乙醇溶液,室温;丙烯酰氯,N,N-二异丙基乙胺,二氯甲烷,室温;(f)三氟乙酸,二氯甲烷,室温;四氢呋喃溶液,氢氧化钠溶液,室温;
其中R1、R2的定义同前所述。
本发明技术方案的另一方面是提供药物组合物,其特征在于,所述药物组合物中包含前述的化合物或其药学上可接受的盐以及药学上可接受的载体。
本发明技术方案的另一方面是提供如通式(I)所示的化合物或其药学上可接受的盐在制备治疗与BTK功能相关的疾病的药物中的应用。
本发明技术方案的另一方面是提供如通式(I)所示的化合物或其药学上可接受的盐在制备治疗类风湿性关节炎,系统性红斑狼疮的药物中的应用。
本发明技术方案的另一方面是提供如通式(I)所示的化合物或其药学上可接受的盐在制备治疗非霍奇金性淋巴瘤的药物中的用途,所述的非霍奇金性淋巴瘤优选套细胞淋巴瘤,弥漫大B细胞淋巴瘤。
本发明技术方案的另一方面是提供如通式(I)所示的化合物或其药学上可接受的盐在制备治疗华式巨球蛋白血症,慢性淋巴细胞白血病或原发性中枢神经系统淋巴瘤药物中的用途。
有益技术效果
本发明技术方案中的所有实施例化合物均对BTK有明显的抑制活性,对TMD-8淋巴瘤细胞均有明显的抑制作用,且实施例8有较高的口服生物利用度,优于上市药物依鲁替尼,并在TMD-8动物模型上展现了较强的肿瘤抑制活性,展现了良好的成药前景。
附图说明
图1化合物的体内抑瘤效果。(A)小鼠体重-时间曲线;(B)肿瘤体积-时间曲线;(C)瘤重-剂量图;(D)各组肿瘤的照片
具体实施方式
实施例1
N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺
(1)N-(4-溴-2-氟苄基)-4-(叔丁基)苯甲酰胺的制备
将2-氟-4-溴苄胺(214mg,1mmol)加入瓶中,用2mL二氯甲烷溶解,加入二异丙基乙胺(194μL,1.1mmol),搅拌均匀,冰浴条件下,滴加4-叔丁基苯甲酰氯(205μL,1.05mmol),室温搅拌1.5h。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=10:1,得白色固体331mg,收率90.93%,熔点101-102℃。
1H NMR(400MHz,Chloroform-d)δ7.74–7.68(m,2H),7.46–7.41(m,2H),7.32–7.27(m,1H),7.25–7.24(m,1H),7.23(dt,J=3.3,1.3Hz,1H),6.59(s,1H),4.62(d,J=5.9Hz,2H),1.32(s,9H).
MS(ESI)m/z 364.07(M+H)+.
(2)4-(叔丁基)-N-(2-氟-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)苯甲酰胺的制备
将N-(4-溴-2-氟苄基)-4-(叔丁基)苯甲酰胺(4.0572g,0.011mol),联硼酸频那醇酯(4.2499g,0.0165mol)和醋酸钾(3.2849g,0.033mol)加入瓶中,加入40mL二氧六环,室温搅拌均匀,加入Pd(dppf)Cl2(0.8164g,0.0011mol),氩气保护下加热至100℃反应6h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=5:1,得白色固体2.38g,收率52.73%,熔点110-111℃
1H NMR(400MHz,Chloroform-d)δ7.73–7.69(m,2H),7.54(dd,J=7.4,1.1Hz,1H),7.47(dd,J=10.4,1.1Hz,1H),7.44(d,J=2.0Hz,1H),7.42(d,J=1.9Hz,1H),7.39(d,J=7.4Hz,1H),6.53(d,J=5.0Hz,1H),4.70(d,J=5.8Hz,2H),1.33(d,J=4.5Hz,21H).
MS(ESI)m/z 412.25(M+H)+.
(3)4-氯-5-碘-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶的制备
将4-氯-5-碘-7H-吡咯并[2,3-d]嘧啶(7g,0.025mol)加入瓶中,用45mL DMF溶解,加入碳酸钾(10.3658g,0.075mol),室温搅拌0.5h,然后冰浴条件下滴加SEMCl(6.6mL,0.0375mol),室温搅拌6h。加水,减压抽滤,收集滤饼,柱层析分离,洗脱剂为石油醚:乙酸乙酯=10:1,得白色固体7.92g,收率77.34%,熔点90-91℃
1H NMR(500MHz,DMSO-d6)δ8.70(s,1H),8.15(s,1H),5.61(s,2H),3.52(t,J=8.0Hz,2H),0.82(t,J=8.0Hz,2H),-0.09(s,9H).
MS(ESI)m/z 410.99(M+H)+.
(4)4-(4-氯)叔丁基-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐的制备
将4-氯-5-碘-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶(1.2406g,3.0278mmol),4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-3,6-二氢吡啶-1(2H)-羧酸叔丁酯(0.9652g,3.0278mmol)和碳酸钾(0.8357g,6.0556mmol)加入瓶中,加入12mL二氧六环和6mL水,室温搅拌均匀,加入Pd(dppf)Cl2(0.2215g,0.30278mmol),氩气保护下加热至100℃反应2h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=5:1,得浅黄色固体1.01g,收率71.63%,熔点75-76℃
1H NMR(500MHz,DMSO-d6)δ8.66(s,1H),7.79(s,1H),5.83(s,1H),5.61(s,2H),3.99(s,2H),3.61–3.49(m,4H),2.44(s,2H),1.43(s,9H),0.82(t,J=8.0Hz,2H),-0.11(s,9H).
MS(ESI)m/z 465.21(M+H)+.
(5)叔丁基4-(4-(4-((4-(叔丁基)苯甲酰氨基)甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐的制备
将4-(4-氯)叔丁基-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(500mg,1.075mmol),4-(叔丁基)-N-(2-氟-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)苯甲酰胺(663mg,1.6125mmol)和碳酸钾(297mg,2.15mmol)加入瓶中,加入5mL二氧六环和2.5mL水,室温搅拌均匀,加入Pd(dppf)Cl2(78mg,0.1075mmol),氩气保护下加热至100℃反应7h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=5:1,得浅黄色固体360mg,收率46.94%,熔点85-86℃。
1H NMR(400MHz,DMSO-d6)δ9.01(t,J=5.7Hz,1H),8.91(s,1H),7.87–7.81(m,2H),7.77(s,1H),7.51–7.45(m,4H),7.41(d,J=10.9Hz,1H),5.65(s,2H),5.39–5.27(m,1H),4.59(d,J=5.8Hz,2H),3.73(s,2H),3.60–3.54(m,2H),3.21(t,J=5.6Hz,2H),1.94(s,2H),1.40(s,9H),1.30(s,9H),0.84(dd,J=8.5,7.5Hz,2H),-0.09(s,9H).
MS(ESI)m/z 714.38(M+H)+.
(6)N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-)吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰氨基)甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(125mg,0.175mmol)加入瓶中,加入0.75mL无水乙醇,室温搅拌均匀,滴加浓盐酸(438μL,5.2mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于1.5mL二氯甲烷,在冰盐浴的条件下加入DIEA(96μL,0.54mmol)和NaOH(10mg,0.25mmol),滴加2-丁炔酰氯(28μL,0.324mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体42mg,收率34.43%,熔点90-91℃。
1H NMR(400MHz,DMSO-d6)δ8.99(dt,J=11.9,5.8Hz,1H),8.92(s,1H),7.84(dd,J=8.4,5.8Hz,2H),7.78(d,J=8.0Hz,1H),7.48(dd,J=9.8,3.3Hz,3H),7.45–7.38(m,2H),5.66(s,2H),5.34(s,1H),4.59(d,J=5.8Hz,2H),4.05(s,1H),3.83(s,1H),3.58(t,J=7.9Hz,2H),3.53(t,J=5.7Hz,1H),3.37(t,J=5.8Hz,1H),2.04(s,1H),2.00(s,3H),1.94(s,1H),1.30(s,9H),0.88–0.81(m,2H),-0.08(s,9H).
MS(ESI)m/z 680.35(M+H)+.
(7)标题化合物的制备
将N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-)吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺(36mg,0.053mmol)溶于600μL二氯甲烷,在冰浴条件下滴加三氟乙酸(280μL,3.71mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得黄色固体20mg,收率68.96%,熔点161-162℃。
1H NMR(400MHz,DMSO-d6)δ12.33(s,1H),8.98(s,1H),8.83(s,1H),7.84(s,2H),7.58(s,1H),7.47(s,3H),7.42(s,2H),5.30(s,1H),4.59(d,J=5.7Hz,2H),4.03(d,J=10.2Hz,1H),3.82(s,1H),3.52(t,J=5.6Hz,1H),2.04(s,1H),1.99(s,3H),1.95(s,1H),1.30(s,9H).
HRMS calcd.For C33H33O2N5F(M+H)+550.2613,found 550.2653.
实施例2
N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺
(1)N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰氨基)甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(125mg,0.175mmol)加入瓶中,加入0.75mL无水乙醇,室温搅拌均匀,滴加浓盐酸(438μL,5.2mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于1.5mL二氯甲烷,在冰盐浴的条件下加入DIEA(192μL,1.08mmol)和NaOH(10mg,0.25mmol),滴加丙烯酰氯(33μL,0.405mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体64mg,收率53.23%,熔点155-158℃。
1H NMR(400MHz,DMSO-d6)δ9.01–8.90(m,1H),8.86(s,1H),7.82–7.68(m,3H),7.45–7.38(m,4H),7.37–7.31(m,1H),6.65(dd,J=16.7,10.4Hz,1H),6.06(dd,J=15.7,11.4Hz,1H),5.60(d,J=8.3Hz,3H),5.27(d,J=72.6Hz,1H),4.52(d,J=5.7Hz,2H),3.90–3.80(m,2H),3.52(t,J=7.9Hz,2H),3.44–3.30(m,2H),1.92(d,J=12.0Hz,2H),1.24(s,9H),0.79(t,J=7.9Hz,2H),-0.14(s,9H).
MS(ESI)m/z 668.34(M+H)+.
(2)标题化合物的制备
将N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺(35mg,0.053mmol)溶于600μL二氯甲烷,在冰浴条件下滴加三氟乙酸(280μL,3.71mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得黄色固体23mg,收率80.72%,熔点179-180℃。
1H NMR(400MHz,DMSO-d6)δ12.39(s,1H),9.09(s,1H),8.82(s,1H),7.85(d,J=7.4Hz,2H),7.59(d,J=20.0Hz,1H),7.51–7.34(m,5H),6.69(dd,J=16.6,10.5Hz,1H),6.21–5.99(m,1H),5.65(t,J=10.2Hz,1H),5.27(d,J=83.4Hz,1H),4.57(d,J=5.4Hz,2H),3.90(s,2H),3.45(s,1H),3.37(s,1H),2.03(s,1H),1.95(s,1H),1.29(s,9H).
HRMS calcd.For C32H33O2N5F(M+H)+538.2613,found 538.2611.
实施例3
N-(4-(5-(1-(丁-2-炔酰基))-1,2,3,6-四氢吡啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)苄基)-4-(叔丁基)苯甲酰胺
(1)N-(4-溴苄基)-4-(叔丁基)苯甲酰胺的制备
将4-溴苄胺(1g,5.375mmol)加入瓶中,用20mL二氯甲烷溶解,加入二异丙基乙胺(1mL,5.9125mmol),搅拌均匀,冰浴条件下,滴加4-叔丁基苯甲酰氯(1.125mL,5.64375mmol),室温搅拌3h。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=10:1,得白色固体1.8g,收率96.72%,熔点110-111℃。
1H NMR(400MHz,Chloroform-d)δ7.75–7.70(m,2H),7.48–7.42(m,4H),7.24–7.19(m,2H),6.43(s,1H),4.59(d,J=5.7Hz,2H),1.33(s,9H).
MS(ESI)m/z 346.98(M+H)+.
(2)4-(叔丁基)-N-(4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)苯甲酰胺的制备
将N-(4-溴苄基)-4-(叔丁基)苯甲酰胺(1.77g,5.112mmol),联硼酸频那醇酯(1.9472g,7.668mmol)和醋酸钾(1.5051g,15.336mmol)加入瓶中,加入20mL二氧六环,室温搅拌均匀,加入Pd(dppf)Cl2(0.374g,0.5112mmol),氩气保护下加热至100℃反应6h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=5:1,得白色固体1.01g,收率50.39%,熔点131-132℃。1H NMR(400MHz,Chloroform-d)δ7.82–7.77(m,2H),7.75–7.69(m,2H),7.47–7.41(m,2H),7.38–7.32(m,2H),6.36(s,1H),4.68–4.65(m,2H),1.34(s,12H),1.33(s,9H).
MS(ESI)m/z 394.24(M+H)+.
(3)叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐的制备
将4-(4-氯)叔丁基-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(500mg,1.075mmol),4-(叔丁基)-N-(4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)苯甲酰胺(634mg,1.6125mmol)和碳酸钾(297mg,2.15mmol)加入瓶中,加入5mL二氧六环和2.5mL水,室温搅拌均匀,加入Pd(dppf)Cl2(78mg,0.1075mmol),氩气保护下加热至100℃反应10h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=5:1,得白色固体610mg,收率81.55%,熔点161-162℃。
1H NMR(500MHz,DMSO-d6)δ9.01(d,J=5.4Hz,1H),8.88(s,1H),7.84(d,J=8.0Hz,2H),7.72(s,1H),7.61(d,J=7.8Hz,2H),7.48(d,J=8.0Hz,2H),7.44(d,J=8.0Hz,2H),5.65(s,2H),5.34(s,1H),4.56(d,J=5.9Hz,2H),3.72(s,2H),3.58(t,J=7.9Hz,2H),3.16(t,J=5.7Hz,2H),1.88(s,2H),1.40(s,9H),1.30(s,9H),0.84(t,J=8.0Hz,2H),-0.06–-0.10(m,9H).
MS(ESI)m/z 696.35(M+H)+.
(4)N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-)吡咯并[2,3-d]嘧啶-4-基)苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(150mg,0.216mmol)加入瓶中,加入0.9mL无水乙醇,室温搅拌均匀,滴加浓盐酸(525μL,6.24mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于2mL二氯甲烷,在冰盐浴的条件下加入DIEA(115μL,0.648mmol)和NaOH(12mg,0.3mmol),滴加2-丁炔酰氯(34μL,0.389mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体57mg,收率40.21%,熔点95-96℃。
1H NMR(500MHz,DMSO-d6)δ8.96(dt,J=11.7,6.0Hz,1H),8.87(s,1H),7.82(t,J=7.6Hz,2H),7.70(d,J=7.4Hz,1H),7.57(t,J=6.5Hz,2H),7.46(d,J=8.0Hz,2H),7.41(t,J=7.0Hz,2H),5.63(s,2H),5.33(s,1H),4.55(d,J=5.8Hz,2H),4.03(s,1H),3.81(s,1H),3.56(t,J=8.0Hz,2H),3.46(t,J=5.4Hz,1H),3.30(s,1H),1.99(d,J=28.1Hz,3H),1.90(s,1H),1.83(s,1H),1.28(d,J=2.2Hz,9H),0.82(t,J=7.9Hz,2H),-0.11(s,9H).
MS(ESI)m/z 662.3497(M+H)+.
(5)标题化合物的制备
将N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-)吡咯并[2,3-d]嘧啶-4-基)苄基)-4-(叔丁基)苯甲酰胺(85mg,0.1277mmol)溶于1.5mL二氯甲烷,在冰浴条件下滴加三氟乙酸(664μL,8.939mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得黄色固体60mg,收率88.37%,熔点139-140℃
1H NMR(400MHz,DMSO-d6)δ12.27(s,1H),9.04–8.94(m,1H),8.81(s,1H),7.86–7.81(m,2H),7.59(dd,J=8.1,3.8Hz,2H),7.55(dd,J=7.3,2.4Hz,1H),7.48(d,J=8.5Hz,2H),7.42(dd,J=8.1,4.4Hz,2H),5.31(d,J=3.2Hz,1H),4.56(d,J=5.9Hz,2H),4.04(s,1H),3.85–3.78(m,1H),3.48(t,J=5.6Hz,1H),2.04(s,1H),1.98(s,3H),1.93(s,1H),1.30(s,9H).
HRMS calcd.For C33H34O2N5(M+H)+532.2707,found 532.2696.
实施例4
N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基))-7H-吡咯并[2,3-d]嘧啶-4-基)苄基)-4-(叔丁基)苯甲酰胺
(1)N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(150mg,0.216mmol)加入瓶中,加入0.9mL无水乙醇,室温搅拌均匀,滴加浓盐酸(525μL,6.24mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于2mL二氯甲烷,在冰盐浴的条件下加入DIEA(230μL,1.296mmol)和NaOH(12mg,0.3mmol),滴加丙烯酰氯(39.6μL,0.486mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体68mg,收率48.52%,熔点71-72℃。
1H NMR(500MHz,DMSO-d6)δ9.01–8.93(m,1H),8.86(s,1H),7.81(d,J=7.9Hz,2H),7.71(d,J=22.4Hz,1H),7.57(t,J=8.7Hz,2H),7.44(d,J=7.9Hz,2H),7.39(t,J=9.0Hz,2H),6.65(dd,J=17.1,10.5Hz,1H),6.16–6.03(m,1H),5.70–5.58(m,3H),5.28(d,J=105.0Hz,1H),4.52(d,J=5.9Hz,2H),3.88(d,J=3.3Hz,2H),3.55(t,J=7.9Hz,2H),3.39(t,J=5.3Hz,1H),3.31(t,J=4.6Hz,1H),1.94(s,1H),1.86(s,1H),1.27(s,9H),0.82(t,J=7.9Hz,2H),-0.11(s,9H).
MS(ESI)m/z 650.38(M+H)+.
(2)标题化合物的制备
将N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)苄基)-4-(叔丁基)苯甲酰胺(83mg,0.1277mmol)溶于1.5mL二氯甲烷,在冰浴条件下滴加三氟乙酸(664μL,8.939mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体62mg,收率93.43%,熔点132-133℃。
1H NMR(500MHz,DMSO-d6)δ12.26(s,1H),9.04–8.97(m,1H),8.80(s,1H),7.83(d,J=8.3Hz,2H),7.61–7.51(m,3H),7.50–7.44(m,2H),7.40(t,J=9.4Hz,2H),6.75–6.62(m,1H),6.11(dd,J=22.8,14.8Hz,1H),5.66(dd,J=25.7,11.4Hz,1H),5.26(d,J=113.8Hz,1H),4.57–4.52(m,2H),3.89(s,2H),3.45–3.40(m,1H),3.33(t,J=5.2Hz,1H),1.99(s,1H),1.89(s,1H),1.30(s,9H).
HRMS calcd.For C32H34O2N5(M+H)+520.2707,found 520.2746.
实施例5
N-(4-(5-(1-(丁-2-炔酰基))-1,2,3,6-四氢吡啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺
(1)N-(4-溴-2-甲基苄基)-4-(叔丁基)苯甲酰胺的制备
将2-甲基-4-溴苄胺(200mg,1mmol)加入瓶中,用2mL二氯甲烷溶解,加入二异丙基乙胺(194μL,1.1mmol),搅拌均匀,冰浴条件下,滴加4-叔丁基苯甲酰氯(205μL,1.05mmol),室温搅拌1.5h。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=10:1,得白色固体333mg,收率92.55%,熔点125-126℃。
1H NMR(400MHz,Chloroform-d)δ7.79–7.63(m,2H),7.49–7.37(m,2H),7.29(ddt,J=19.4,11.5,4.5Hz,2H),7.13(d,J=8.2Hz,1H),6.29(s,1H),4.55(d,J=5.5Hz,2H),2.31(s,3H),1.31(s,9H).
MS(ESI)m/z 361.06(M+H)+.
(2)4-(叔丁基)-N-(2-甲基-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)苯甲酰胺的制备
将N-(4-溴-2-甲基苄基)-4-(叔丁基)苯甲酰胺(1.84g,5.112mmol),联硼酸频那醇酯(1.9472g,7.668mmol)和醋酸钾(1.5051g,15.336mmol)加入瓶中,加入20mL二氧六环,室温搅拌均匀,加入Pd(dppf)Cl2(0.374g,0.5112mmol),氩气保护下加热至100℃反应6h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=5:1,得白色固体1.02g,收率48.87%,熔点133-134℃。1H NMR(400MHz,Chloroform-d)δ7.71–7.67(m,2H),7.65–7.59(m,2H),7.45–7.40(m,2H),7.29(d,J=7.5Hz,1H),6.20(s,1H),4.64(d,J=5.3Hz,2H),2.35(s,3H),1.33(s,12H),1.31(s,9H).
MS(ESI)m/z 408.27(M+H)+.
(3)叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)-3-甲基苯基)-7-((2-(三甲基甲硅烷基))乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐的制备
将4-(4-氯)叔丁基-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(500mg,1.075mmol),4-(叔丁基)-N-(2-甲基-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)苯甲酰胺(656mg,1.6125mmol)和碳酸钾(297mg,2.15mmol)加入瓶中,加入5mL二氧六环和2.5mL水,室温搅拌均匀,加入Pd(dppf)Cl2(78mg,0.1075mmol),氩气保护下加热至100℃反应10h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=5:1,得白色固体524mg,收率68.65%,熔点86-87℃。
1H NMR(400MHz,DMSO-d6)δ8.94–8.89(m,1H),8.88(s,1H),7.85(d,J=8.2Hz,2H),7.74(s,1H),7.53(d,J=7.7Hz,1H),7.49(d,J=8.2Hz,2H),7.41(s,1H),7.38(d,J=7.9Hz,1H),5.65(s,2H),5.34(s,1H),4.53(d,J=5.7Hz,2H),3.72(s,2H),3.57(t,J=7.9Hz,2H),3.17(t,J=5.6Hz,2H),2.38(s,3H),1.90(s,2H),1.40(s,9H),1.30(s,9H),0.85(dd,J=10.0,5.9Hz,2H),-0.09(s,9H).
MS(ESI)m/z 710.41(M+H)+.
(4)N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-)吡咯并[2,3-d]嘧啶-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)-3-甲基苯基)-7-((2-(三甲基甲硅烷基))乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(153mg,0.216mmol)加入瓶中,加入0.9mL无水乙醇,室温搅拌均匀,滴加浓盐酸(525μL,6.24mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于2mL二氯甲烷,在冰盐浴的条件下加入DIEA(115μL,0.648mmol)和NaOH(12mg,0.3mmol),滴加2-丁炔酰氯(34μL,0.389mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体130mg,收率89.04%,熔点84-85℃。
1H NMR(500MHz,DMSO-d6)δ8.89–8.78(m,2H),7.82(t,J=7.9Hz,2H),7.70(d,J=13.2Hz,1H),7.44(dd,J=10.3,7.7Hz,3H),7.34(dd,J=19.6,8.2Hz,2H),5.61(s,2H),5.31(s,1H),4.50(d,J=5.7Hz,2H),4.01(s,1H),3.86–3.70(m,1H),3.54(t,J=7.9Hz,2H),3.47(t,J=5.7Hz,1H),3.31(d,J=11.5Hz,1H),2.34(d,J=4.4Hz,3H),1.99(d,J=29.7Hz,3H),1.89(d,J=14.7Hz,2H),1.27(s,9H),0.81(d,J=6.4Hz,2H),-0.12(s,9H).
MS(ESI)m/z 676.36(M+H)+.
(5)标题化合物的制备
将N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-)吡咯并[2,3-d]嘧啶-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺(86mg,0.1277mmol)溶于1.5mL二氯甲烷,在冰浴条件下滴加三氟乙酸(664μL,8.939mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体59mg,收率84.67%,熔点162-163℃。
1H NMR(400MHz,DMSO-d6)δ12.21(s,1H),8.87–8.78(m,1H),8.75(s,1H),7.81(dd,J=8.3,6.0Hz,2H),7.51(d,J=12.2Hz,1H),7.43(t,J=8.7Hz,3H),7.32(dd,J=17.7,8.5Hz,2H),5.24(d,J=9.6Hz,1H),4.48(d,J=5.6Hz,2H),3.98(s,1H),3.78(s,1H),3.45(t,J=5.4Hz,1H),3.30(d,J=4.9Hz,1H),2.32(s,3H),1.94(s,2H),1.90(s,3H),1.26(s,9H).
HRMS calcd.For C34H36O2N5(M+H)+546.2864,found 546.2859.
实施例6
N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺
(1)N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)-3-甲基苯基)-7-((2-(三甲基甲硅烷基))乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(153mg,0.216mmol)加入瓶中,加入0.9mL无水乙醇,室温搅拌均匀,滴加浓盐酸(525μL,6.24mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于2mL二氯甲烷,在冰盐浴的条件下加入DIEA(230μL,1.296mmol)和NaOH(12mg,0.3mmol),滴加丙烯酰氯(39.6μL,0.486mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体98mg,收率68.33%,熔点90-91℃。
1H NMR(500MHz,DMSO-d6)δ8.88(s,2H),7.85(d,J=8.0Hz,2H),7.75(d,J=17.1Hz,1H),7.48(t,J=6.5Hz,3H),7.41(s,1H),7.35(d,J=7.4Hz,1H),6.70(dt,J=16.5,10.7Hz,1H),6.12(t,J=19.2Hz,1H),5.65(s,3H),5.35(d,J=79.3Hz,1H),4.51(d,J=5.7Hz,2H),3.92(d,J=10.1Hz,2H),3.57(t,J=8.0Hz,2H),3.44–3.35(m,2H),2.35(s,3H),1.96(s,1H),1.90(s,1H),1.30(s,10H),0.86–0.82(m,2H),-0.09(s,9H).
MS(ESI)m/z 664.37(M+H)+.
(2)标题化合物的制备
将N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺(85mg,0.1277mmol)溶于1.5mL二氯甲烷,在冰浴条件下滴加三氟乙酸(664μL,8.939mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体50mg,收率73.37%,熔点165-166℃。
1H NMR(400MHz,DMSO-d6)δ12.21(d,J=9.9Hz,1H),8.84(t,J=5.5Hz,1H),8.74(s,1H),7.79(d,J=8.0Hz,2H),7.51(d,J=13.6Hz,1H),7.42(d,J=7.2Hz,3H),7.35(s,1H),7.29(d,J=7.3Hz,1H),6.64(dt,J=16.6,9.4Hz,1H),6.06(t,J=16.2Hz,1H),5.60(t,J=9.4Hz,1H),5.24(d,J=68.0Hz,1H),4.46(d,J=5.7Hz,2H),3.85(s,2H),3.36(s,1H),3.11(s,1H),2.30(s,3H),1.89(d,J=28.3Hz,2H),1.24(s,9H).
HRMS calcd.For C33H36O2N5(M+H)+534.2864,found 534.2849.
实施例7
N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶)-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)苯甲酰胺
(1)N-(4-溴-2-氟苄基)苯甲酰胺的制备
将2-氟-4-溴苄胺(700mg,3.43mmol)加入瓶中,用5mL二氯甲烷溶解,加入二异丙基乙胺(660μL,3.773mmol),搅拌均匀,冰浴条件下,滴加苯甲酰氯(410μL,3.6mmol),室温搅拌6.5h。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=10:1,得白色固体982mg,收率92.90%,熔点78-79℃。
1H NMR(400MHz,DMSO-d6)δ9.04(t,J=5.8Hz,1H),7.92–7.85(m,2H),7.57–7.51(m,2H),7.50–7.44(m,2H),7.40(dd,J=8.2,1.9Hz,1H),7.32(t,J=8.1Hz,1H),4.47(d,J=5.8Hz,2H).
MS(ESI)m/z 308.01(M+H)+.
(2)N-(2-氟-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)苯甲酰胺的制备
将N-(4-溴-2-氟苄基)苯甲酰胺(982mg,3.18676mmol),联硼酸频那醇酯(1.214g,4.78mmol)和醋酸钾(938mg,9.56mmol)加入瓶中,加入10mL二氧六环,室温搅拌均匀,加入Pd(dppf)Cl2(233mg,0.318676mmol),氩气保护下加热至100℃反应6h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=5:1,得白色固体517mg,收率45.67%,熔点110-111℃。
1H NMR(500MHz,DMSO-d6)δ9.09–8.99(m,1H),7.90(q,J=6.7,6.1Hz,2H),7.60–7.44(m,4H),7.43–7.28(m,2H),4.55(q,J=7.0,6.4Hz,2H),1.30(dd,J=9.7,4.7Hz,12H).
MS(ESI)m/z 356.08(M+H)+.
(3)叔丁基4-(4-(4-(苯甲酰胺甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐的制备
将4-(4-氯)叔丁基-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(400mg,0.86mmol),N-(2-氟-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)苯甲酰胺(458mg,1.29mmol)和碳酸钾(237mg,1.72mmol)加入瓶中,加入4mL二氧六环和2mL水,室温搅拌均匀,加入Pd(dppf)Cl2(63mg,0.086mmol),氩气保护下加热至100℃反应10h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=5:1,得黄色固体320mg,收率56.56%,熔点81-82℃。
1H NMR(500MHz,DMSO-d6)δ9.04(s,1H),8.88(s,1H),7.87(d,J=7.7Hz,2H),7.74(s,1H),7.54–7.36(m,7H),5.62(s,2H),5.29(d,J=16.7Hz,1H),4.57(d,J=5.8Hz,2H),3.69(s,2H),3.56(q,J=9.5,7.8Hz,2H),3.42(q,J=6.6Hz,2H),1.91(s,2H),1.37(s,9H),0.82(t,J=7.9Hz,2H),-0.11(s,9H).
MS(ESI)m/z 658.33(M+H)+.
(4)N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-)吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)苯甲酰胺的制备
将叔丁基4-(4-(4-(苯甲酰胺甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(160mg,0.243mmol)加入瓶中,加入1.5mL无水乙醇,室温搅拌均匀,滴加浓盐酸(2mL,24mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于2mL二氯甲烷,在冰盐浴的条件下加入DIEA(130μL,0.729mmol)和NaOH(12mg,0.3mmol),滴加2-丁炔酰氯(38μL,0.44mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得黄色固体76mg,收率50.14%,熔点82-83℃。
1H NMR(500MHz,DMSO-d6)δ9.05(d,J=13.7Hz,1H),8.91(s,1H),7.90(t,J=7.1Hz,2H),7.78(d,J=11.7Hz,1H),7.47(tt,J=30.9,8.4Hz,6H),5.66(s,2H),5.35(d,J=9.9Hz,1H),4.60(d,J=5.8Hz,2H),4.05(s,1H),3.84(s,1H),3.58(t,J=7.9Hz,2H),3.53(s,1H),3.41–3.36(m,1H),1.99(s,3H),1.95(s,1H),1.24(s,1H),0.85(t,J=7.8Hz,2H),-0.08(s,9H).
MS(ESI)m/z 624.33(M+H)+.
(5)标题化合物的制备
将N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-)吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)苯甲酰胺(76mg,0.122mmol)溶于2mL二氯甲烷,在冰浴条件下滴加三氟乙酸(815μL,10.98mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体36mg,收率59.85%,熔点146-147℃。
1H NMR(500MHz,DMSO-d6)δ12.33(s,1H),9.04(dd,J=12.7,6.3Hz,1H),8.83(s,1H),7.90(t,J=6.8Hz,2H),7.62–7.52(m,2H),7.50–7.37(m,5H),5.30(d,J=10.9Hz,1H),4.60(d,J=5.9Hz,2H),4.05(s,1H),3.83(s,1H),3.53(t,J=5.7Hz,1H),3.38(t,J=5.3Hz,1H),2.04(s,1H),1.99(s,4H).
HRMS calcd.For C29H25O2N5F(M+H)+494.1987,found 494.2026.
实施例8
N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)苯甲酰胺
(1)N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)-2-氟苄基)苯甲酰胺的制备
将叔丁基4-(4-(4-(苯甲酰胺甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(160mg,0.243mmol)加入瓶中,加入1.5mL无水乙醇,室温搅拌均匀,滴加浓盐酸(2mL,24mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于2mL二氯甲烷,在冰盐浴的条件下加入DIEA(130μL,0.729mmol)和NaOH(12mg,0.3mmol),滴加丙烯酰氯(45μL,0.55mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体50mg,收率33.78%,
熔点63-64℃。
1H NMR(500MHz,DMSO-d6)δ9.12–9.03(m,1H),8.91(s,1H),7.90(d,J=7.5Hz,2H),7.78(d,J=21.6Hz,1H),7.57–7.38(m,6H),6.69(dd,J=16.6,10.4Hz,1H),6.11(t,J=15.3Hz,1H),5.65(d,J=11.6Hz,3H),5.43–5.20(m,1H),4.58(d,J=5.8Hz,2H),3.91(d,J=12.3Hz,2H),3.58(t,J=8.0Hz,2H),3.42(d,J=31.6Hz,2H),2.03–1.96(m,2H),0.85(t,J=7.9Hz,2H),-0.08(s,9H).
MS(ESI)m/z 612.30(M+H)+.
(2)标题化合物的制备
将N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)-2-氟苄基)苯甲酰胺(37mg,0.06mmol)溶于1mL二氯甲烷,在冰浴条件下滴加三氟乙酸(400μL,5.4mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体17mg,收率58.90%,熔点159-160℃。
1H NMR(500MHz,DMSO-d6)δ12.33(d,J=15.6Hz,1H),9.10–9.03(m,1H),8.83(s,1H),7.90(d,J=6.5Hz,2H),7.63–7.50(m,2H),7.50–7.36(m,5H),6.69(dd,J=17.0,9.8Hz,1H),6.15–6.06(m,1H),5.69–5.61(m,1H),5.28(d,J=99.1Hz,1H),4.58(d,J=5.7Hz,2H),3.90(s,2H),3.46(s,1H),3.38(s,1H),2.04(d,J=6.5Hz,1H),1.98(d,J=10.4Hz,1H).
HRMS calcd.For C28H25O2N5F(M+H)+482.1987,found 482.2021.
实施例9
N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-环丙基苯甲酰胺
(1)N-(4-溴-2-氟苄基)-4-环丙基苯甲酰胺的制备
将2-氟-4-溴苄胺(214mg,1mmol)加入瓶中,用2mL二氯甲烷溶解,加入二异丙基乙胺(194μL,1.1mmol),搅拌均匀,冰浴条件下,滴加4-环丙基苯甲酰氯(195mg,1.05mmol),室温搅拌1.5h。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=10:1,得白色固体319mg,收率91.63%。
熔点91-92℃
1H NMR(500MHz,DMSO-d6)δ8.91(s,1H),7.77(d,J=8.0Hz,2H),7.51(d,J=9.8Hz,1H),7.39(d,J=8.4Hz,1H),7.30(t,J=8.2Hz,1H),7.16(d,J=8.1Hz,2H),4.44(d,J=5.7Hz,2H),2.01–1.94(m,1H),1.00(d,J=8.2Hz,2H),0.73(d,J=5.2Hz,2H).
MS(ESI)m/z 348.04(M+H)+.
(2)4-环丙基-N-(2-氟-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)苯甲酰胺的制备
将N-(4-溴-2-氟苄基)-4-环丙基苯甲酰胺(360mg,1.034mmol),联硼酸频那醇酯(394mg,1.551mmol)和醋酸钾(304mg,3.102mmol)加入瓶中,加入5.5mL二氧六环,室温搅拌均匀,加入Pd(dppf)Cl2(76mg,0.1034mmol),氩气保护下加热至100℃反应6h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=5:1,得白色固体320mg,收率78.30%,熔点95-96℃。
1H NMR(500MHz,DMSO-d6)δ8.92(t,J=4.6Hz,1H),7.78(d,J=8.0Hz,2H),7.45(d,J=7.5Hz,1H),7.39–7.29(m,2H),7.16(d,J=8.0Hz,2H),4.51(d,J=5.8Hz,2H),2.01–1.95(m,1H),1.29(s,12H),1.03–0.98(m,2H),0.76–0.71(m,2H).MS(ESI)m/z 396.17(M+H)+.
(3)叔丁基4-(4-(4-((4-环丙基苯甲酰胺基)甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐的制备
将4-(4-氯)叔丁基-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(300mg,0.6423mmol),4-环丙基-N-(2-氟-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)苯甲酰胺(330mg,0.835mmol)和碳酸钾(177mg,1.2846mmol)加入瓶中,加入3mL二氧六环和1.5mL水,室温搅拌均匀,加入Pd(dppf)Cl2(47mg,0.06423mmol),氩气保护下加热至100℃反应10h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=2:1,得白色固体275mg,收率61.32%,熔点65-66℃。
1H NMR(500MHz,DMSO-d6)δ8.96(t,J=5.8Hz,1H),8.91(s,1H),7.81–7.74(m,3H),7.51–7.45(m,2H),7.40(d,J=10.8Hz,1H),7.15(d,J=8.0Hz,2H),5.65(s,2H),5.33(s,1H),4.57(d,J=5.7Hz,2H),3.72(s,2H),3.58(t,J=7.9Hz,2H),3.20(s,2H),1.96(d,J=13.6Hz,2H),1.40(s,9H),1.24(s,1H),1.00(d,J=8.1Hz,2H),0.85(t,J=7.9Hz,2H),0.73(d,J=5.5Hz,2H),-0.08(s,9H).
MS(ESI)m/z 698.30(M+H)+.
(4)N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-)吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-环丙基苯甲酰胺的制备
将叔丁基4-(4-(4-((4-环丙基苯甲酰胺基)甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(211mg,0.303mmol)加入瓶中,加入2.5mL无水乙醇,室温搅拌均匀,滴加浓盐酸(1.2mL,15.15mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于3mL二氯甲烷,在冰盐浴的条件下加入DIEA(160μL,0.909mmol)和NaOH(17mg,0.42mmol),滴加2-丁炔酰氯(50μL,0.5454mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得黄色固体150mg,收率74.63%,熔点67-68℃。
1H NMR(400MHz,DMSO-d6)δ8.96(dt,J=11.5,6.0Hz,1H),8.91(s,1H),7.79(dd,J=8.6,5.3Hz,3H),7.52–7.36(m,3H),7.15(d,J=8.3Hz,2H),5.65(s,2H),5.35(s,1H),4.58(d,J=5.7Hz,2H),4.04(s,1H),3.83(s,1H),3.57(t,J=8.0Hz,2H),3.51(t,J=5.6Hz,1H),3.36(d,J=6.6Hz,1H),2.04(s,1H),2.01–1.90(m,5H),1.04–0.96(m,2H),0.85(t,J=8.0Hz,2H),0.77–0.70(m,2H),-0.08(s,8H).
MS(ESI)m/z 664.33(M+H)+.
(5)标题化合物的制备
将N-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-)吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-环丙基苯甲酰胺(150mg,0.226mmol)溶于3mL二氯甲烷,在冰浴条件下滴加三氟乙酸(1.5mL,20.34mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体96mg,收率79.61%,熔点135-136℃。
1H NMR(400MHz,DMSO-d6)δ12.29(s,1H),8.96–8.87(m,1H),8.78(s,1H),7.74(dd,J=8.4,4.5Hz,2H),7.55(dd,J=10.2,2.4Hz,1H),7.45–7.29(m,3H),7.10(d,J=8.4Hz,2H),5.24(d,J=8.8Hz,1H),4.52(d,J=5.4Hz,2H),3.99(s,1H),3.80–3.71(m,1H),3.46(t,J=5.8Hz,1H),3.32(t,J=5.9Hz,1H),1.99(s,1H),1.96–1.88(m,5H),0.96(dd,J=8.4,2.3Hz,2H),0.69(dd,J=4.9,2.1Hz,2H).
HRMS calcd.For C32H29O2N5F(M+H)+534.2300,found 534.2295.
实施例10
N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-环丙基苯甲酰胺
(1)N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)-2-氟苄基)-4-环丙基苯甲酰胺的制备
将叔丁基4-(4-(4-((4-环丙基苯甲酰胺基)甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(211mg,0.303mmol)加入瓶中,加入2.5mL无水乙醇,室温搅拌均匀,滴加浓盐酸(1.2mL,15.15mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于3mL二氯甲烷,在冰盐浴的条件下加入DIEA(160μL,0.909mmol)和NaOH(17mg,0.42mmol),滴加丙烯酰氯(55μL,0.682mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体124mg,收率62.78%,熔点66-67℃。
1H NMR(400MHz,DMSO-d6)δ9.01–8.94(m,1H),8.91(s,1H),7.83–7.74(m,3H),7.50–7.36(m,3H),7.14(d,J=7.9Hz,2H),6.68(dd,J=16.6,10.5Hz,1H),6.11(dd,J=17.0,9.9Hz,1H),5.65(d,J=8.0Hz,3H),5.32(d,J=77.7Hz,1H),4.56(d,J=5.7Hz,2H),3.90(s,2H),3.57(t,J=8.0Hz,2H),3.48–3.41(m,1H),3.40–3.34(m,1H),2.01(s,1H),1.97(dt,J=8.4,3.7Hz,2H),1.03–0.97(m,2H),0.89–0.81(m,3H),0.76–0.69(m,2H),-0.09(s,9H).
MS(ESI)m/z 652.35(M+H)+.
(2)标题化合物的制备
将N-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)-2-氟苄基)-4-环丙基苯甲酰胺(110mg,0.169mmol)溶于1mL二氯甲烷,在冰浴条件下滴加三氟乙酸(1.13mL,15.21mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体76mg,收率86.31%,熔点135-136℃。
1H NMR(400MHz,DMSO-d6)δ12.34(d,J=13.3Hz,1H),9.01–8.93(m,1H),8.82(s,1H),7.78(d,J=7.3Hz,2H),7.60(d,J=18.4Hz,1H),7.41(d,J=16.9Hz,3H),7.14(dd,J=7.3,3.2Hz,2H),6.68(dd,J=16.5,10.6Hz,1H),6.10(dd,J=17.1,9.0Hz,1H),5.66(t,J=8.1Hz,1H),5.27(d,J=84.0Hz,1H),4.56(d,J=5.7Hz,2H),3.89(s,2H),3.41(dt,J=33.0,4.1Hz,2H),2.03(s,1H),1.95(dd,J=8.0,5.1Hz,2H),1.00(dd,J=8.4,2.2Hz,3H),0.73(dd,J=4.7,1.9Hz,2H).
HRMS calcd.For C31H29O2N5F(M+H)+522.2300,found 522.2291.
实施例11
N-(4-(5-(1-(丁-2-炔酰基)哌啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺
(1)叔丁基4-(4-(4-((4-(叔丁基)苯甲酰氨基)甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)哌啶-1-羧酸盐的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰氨基)甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(410mg,0.574mmol)加入瓶中,溶于16mL甲醇和8mL乙酸乙酯,加入钯碳(230mg,10%),通入氢气,室温下搅拌48h。硅藻土过滤除去Pd/C,浓缩反应液,得白色固体400mg,收率97.32%,熔点87-88℃。
1H NMR(500MHz,DMSO-d6)δ9.04(t,J=6.1Hz,1H),8.85(s,1H),7.83(d,J=8.2Hz,2H),7.59(s,1H),7.53(t,J=7.7Hz,1H),7.50–7.39(m,4H),5.61(s,2H),4.60(d,J=5.8Hz,2H),3.84(s,2H),3.54(t,J=7.9Hz,2H),2.61(t,J=12.1Hz,1H),2.36(s,2H),1.52–1.43(m,2H),1.36(s,9H),1.30(s,9H),1.25–1.18(m,2H),0.82(t,J=7.9Hz,2H),-0.09(s,9H).
MS(ESI)m/z 716.40(M+H)+.
(2)N-(4-(5-(1-(丁-2-炔酰基)哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶)-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰氨基)甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)哌啶-1-羧酸盐(217mg,0.303mmol)加入瓶中,加入2.5mL无水乙醇,室温搅拌均匀,滴加浓盐酸(1.2mL,15.15mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于3mL二氯甲烷,在冰盐浴的条件下加入DIEA(160μL,0.909mmol)和NaOH(17mg,0.42mmol),滴加2-丁炔酰氯(50μL,0.5454mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体131mg,收率63.22%。
熔点82-83℃
1H NMR(400MHz,DMSO-d6)δ9.02(t,J=5.8Hz,1H),8.81(s,1H),7.79–7.74(m,2H),7.55(s,1H),7.50(t,J=7.7Hz,1H),7.44–7.35(m,4H),5.56(s,2H),4.55(d,J=5.8Hz,2H),4.17(d,J=13.1Hz,1H),4.05(d,J=13.1Hz,1H),3.52–3.46(m,2H),2.59(d,J=12.8Hz,2H),2.20(t,J=12.2Hz,1H),1.92(s,3H),1.50(d,J=13.0Hz,2H),1.25(s,9H),1.21–1.19(m,2H),0.81–0.75(m,2H),-0.14(s,9H).
MS(ESI)m/z 682.36(M+H)+.
(3)标题化合物的制备
将N-(4-(5-(1-(丁-2-炔酰基)哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶)-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺(55mg,0.08mmol)溶于1mL二氯甲烷,在冰浴条件下滴加三氟乙酸(420μL,5.646mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体35mg,收率79.30%,熔点156-157℃。
1H NMR(400MHz,DMSO-d6)δ12.12(s,1H),9.08(t,J=5.9Hz,1H),8.77(s,1H),7.82(d,J=8.5Hz,2H),7.57–7.50(m,1H),7.49–7.45(m,2H),7.43–7.38(m,2H),4.60(d,J=5.7Hz,2H),4.23(d,J=13.4Hz,1H),4.11(d,J=15.8Hz,1H),2.71–2.57(m,2H),2.24(s,1H),1.98(s,3H),1.54(d,J=12.4Hz,2H),1.30(s,9H),1.24(s,2H).
HRMS calcd.For C33H35O2N5F(M+H)+552.2769,found 552.2813.
实施例12
N-(4-(5-(1-丙烯酰哌啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺
(1)N-(4-(5-(1-丙烯酰哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰氨基)甲基)-3-氟苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)哌啶-1-羧酸盐(217mg,0.303mmol)加入瓶中,加入2.5mL无水乙醇,室温搅拌均匀,滴加浓盐酸(1.2mL,15.15mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于3mL二氯甲烷,在冰盐浴的条件下加入DIEA(160μL,0.909mmol)和NaOH(17mg,0.42mmol),滴加丙烯酰氯(55μL,0.682mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体118mg,收率58.36%。
熔点79-80℃
1H NMR(500MHz,DMSO-d6)δ9.05(t,J=5.8Hz,1H),8.85(s,1H),7.82(d,J=8.1Hz,2H),7.60–7.52(m,2H),7.44(dd,J=15.9,8.3Hz,4H),6.66(dd,J=16.7,10.4Hz,1H),6.03(d,J=16.8Hz,1H),5.60(d,J=11.0Hz,3H),4.60(d,J=5.8Hz,2H),4.37(d,J=12.8Hz,1H),3.87(d,J=13.5Hz,1H),3.54(t,J=8.0Hz,2H),2.64(dt,J=39.0,13.5Hz,2H),2.25(t,J=13.5Hz,1H),1.53(dd,J=63.0,7.5Hz,2H),1.29(s,9H),1.24(s,2H),0.86–0.79(m,2H),-0.09(s,9H).
MS(ESI)m/z 670.33(M+H)+.
(2)标题化合物的制备
将N-(4-(5-(1-丙烯酰哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-4-(叔丁基)苯甲酰胺(40mg,0.06mmol)溶于1mL二氯甲烷,在冰浴条件下滴加三氟乙酸(400μL,5.4mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体23mg,收率71.04%,熔点154-155℃。
1H NMR(500MHz,DMSO-d6)δ12.05(s,1H),9.02(t,J=5.7Hz,1H),8.73(s,1H),7.79(d,J=8.1Hz,2H),7.51(t,J=7.7Hz,1H),7.45–7.33(m,6H),6.62(dd,J=16.9,10.4Hz,1H),6.00(d,J=15.0Hz,1H),5.56(d,J=12.0Hz,1H),4.57(d,J=5.8Hz,2H),4.34(d,J=13.0Hz,1H),3.83(d,J=10.6Hz,1H),2.63–2.53(m,2H),2.21(s,1H),1.55(d,J=12.6Hz,1H),1.43(d,J=11.9Hz,1H),1.26(s,9H),1.21(s,2H).HRMS calcd.For C32H35O2N5F(M+H)+540.2769,found 540.2809.
实施例13
N-(4-(5-(1-(丁-2-炔酰基)哌啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)苄基)-4-(叔丁基))苯甲酰胺
(1)叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)哌啶-1-羧酸盐的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(290mg,0.4167mmol)加入瓶中,溶于20mL甲醇和10mL乙酸乙酯,加入钯碳(250mg,10%),通入氢气,室温下搅拌48h。硅藻土过滤除去Pd/C,浓缩反应液,得白色固体270mg,收率92.78%,熔点92-93℃。
1H NMR(500MHz,DMSO-d6)δ9.05(t,J=6.1Hz,1H),8.83(s,1H),7.83(d,J=8.2Hz,2H),7.60–7.53(m,3H),7.49(t,J=7.5Hz,4H),5.61(s,2H),4.57(d,J=6.0Hz,2H),3.80(s,2H),3.54(t,J=7.9Hz,2H),2.64(t,J=11.5Hz,1H),2.32(s,2H),1.44(d,J=12.6Hz,2H),1.35(s,9H),1.30(s,9H),1.17(d,J=11.9Hz,2H),0.82(t,J=7.9Hz,2H),-0.10(s,9H).
MS(ESI)m/z 698.38(M+H)+.
(2)N-(4-(5-(1-(丁-2-炔酰基)哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶)-4-基)苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)哌啶-1-羧酸盐(135mg,0.1934mmol)加入瓶中,加入2mL无水乙醇,室温搅拌均匀,滴加浓盐酸(483μL,5.8mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于1.5mL二氯甲烷,在冰盐浴的条件下加入DIEA(100μL,0.58mmol)和NaOH(12mg,0.3mmol),滴加2-丁炔酰氯(30μL,0.348mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体85mg,收率66.41%,熔点96-98℃。
1H NMR(500MHz,DMSO-d6)δ9.05(t,J=6.1Hz,1H),8.84(s,1H),7.82(d,J=8.2Hz,2H),7.60–7.53(m,3H),7.51(d,J=7.8Hz,2H),7.47(d,J=8.2Hz,2H),5.60(s,2H),4.57(d,J=6.0Hz,2H),4.20(t,J=12.3Hz,1H),4.05(dd,J=20.4,10.3Hz,1H),3.54(t,J=7.9Hz,2H),2.66(dt,J=25.6,12.3Hz,2H),2.22(t,J=12.7Hz,1H),1.97(s,3H),1.52(d,J=12.8Hz,2H),1.30(s,9H),1.24(s,2H),0.85–0.80(m,2H),-0.09(s,9H).
MS(ESI)m/z 664.37(M+H)+.
(3)标题化合物的制备
将N-(4-(5-(1-(丁-2-炔酰基)哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶)-4-基)苄基)-4-(叔丁基)苯甲酰胺(75mg,0.113mmol)溶于1.5mL二氯甲烷,在冰浴条件下滴加三氟乙酸(760μL,10.2mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体43mg,收率71.30%,熔点156-157℃。
1H NMR(400MHz,DMSO-d6)δ12.06(s,1H),9.07(t,J=6.1Hz,1H),8.76(s,1H),7.82(d,J=8.4Hz,2H),7.57(d,J=7.9Hz,2H),7.49(dd,J=10.8,8.4Hz,4H),7.37(s,1H),4.57(d,J=6.0Hz,2H),4.20(d,J=13.5Hz,1H),4.08(d,J=12.5Hz,1H),2.67–2.60(m,2H),2.26–2.16(m,1H),1.97(s,3H),1.51(d,J=12.0Hz,2H),1.31(s,9H),1.25(s,2H).
HRMS calcd.For C33H36O2N5(M+H)+534.2864,found 534.2904.
实施例14
N-(4-(5-(1-丙烯酰基哌啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)苄基)-4-(叔丁基)苯甲酰胺
(1)N-(4-(5-(1-丙烯酰哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)哌啶-1-羧酸盐(135mg,0.1934mmol)加入瓶中,加入2mL无水乙醇,室温搅拌均匀,滴加浓盐酸(483μL,5.8mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于1.5mL二氯甲烷,在冰盐浴的条件下加入DIEA(200μL,1.16mmol)和NaOH(12mg,0.3mmol),滴加丙烯酰氯(35μL,0.435mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体64mg,收率50.79%,熔点82-83℃。
1H NMR(400MHz,DMSO-d6)δ9.02(t,J=6.1Hz,1H),8.79(s,1H),7.79–7.73(m,2H),7.53(d,J=8.1Hz,2H),7.50–7.44(m,3H),7.43–7.37(m,2H),6.60(dd,J=16.7,10.5Hz,1H),5.97(dd,J=16.7,2.4Hz,1H),5.56(s,2H),5.53(d,J=2.5Hz,1H),4.52(d,J=6.0Hz,2H),4.28(d,J=12.7Hz,1H),3.78(d,J=13.3Hz,1H),3.54–3.43(m,2H),2.66(t,J=12.0Hz,1H),2.53(t,J=12.9Hz,1H),2.18(t,J=12.5Hz,1H),1.51(d,J=13.0Hz,1H),1.38(d,J=11.8Hz,1H),1.24(s,9H),1.10(d,J=13.3Hz,2H),0.80–0.73(m,2H),-0.15(s,9H).
MS(ESI)m/z 652.37(M+H)+.
(2)标题化合物的制备
将N-(4-(5-(1-丙烯酰哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)苄基)-4-(叔丁基)苯甲酰胺(64mg,0.1mmol)溶于1.5mL二氯甲烷,在冰浴条件下滴加三氟乙酸(670μL,9mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体37mg,收率70.92%。
熔点152-153℃
1H NMR(400MHz,DMSO-d6)δ11.98(s,1H),9.02(t,J=6.0Hz,1H),8.70(s,1H),7.77(d,J=8.5Hz,2H),7.52(d,J=8.1Hz,2H),7.43(dd,J=15.2,8.3Hz,4H),7.30(d,J=2.1Hz,1H),6.59(dd,J=16.7,10.5Hz,1H),5.97(dd,J=16.7,2.4Hz,1H),5.54(dd,J=10.5,2.4Hz,1H),4.52(d,J=5.9Hz,2H),4.29(d,J=12.6Hz,1H),3.78(d,J=14.6Hz,1H),2.68–2.51(m,2H),2.21–2.12(m,1H),1.51(d,J=13.9Hz,1H),1.38(d,J=13.2Hz,1H),1.24(s,9H),1.20(s,2H).
HRMS calcd.For C32H36O2N5(M+H)+522.2864,found 522.2903.
实施例15
N-(4-(5-(1-(丁-2-炔酰基)哌啶-4-基))-7H-吡咯并[2,3-d]嘧啶-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺
(1)叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)-3-甲基苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)哌啶-1-羧酸盐的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)-3-甲基苯基)-7-((2-(三甲基甲硅烷基))乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(450mg,0.6338mmol)加入瓶中,溶于20mL甲醇和10mL乙酸乙酯,加入钯碳(500mg,10%),通入氢气,室温下搅拌48h。硅藻土过滤除去Pd/C,浓缩反应液,得白色固体420mg,收率93.66%,熔点92-93℃。
1H NMR(400MHz,DMSO-d6)δ8.94(t,J=5.9Hz,1H),8.82(s,1H),7.89–7.81(m,2H),7.56(d,J=0.8Hz,1H),7.51–7.46(m,2H),7.42(dd,J=9.0,1.2Hz,3H),5.61(s,2H),4.55(d,J=5.9Hz,2H),3.83(s,2H),3.53(dd,J=8.4,7.5Hz,2H),2.65(t,J=11.6Hz,1H),2.45(s,3H),2.40–2.20(m,2H),1.52–1.43(m,2H),1.35(s,9H),1.30(s,9H),1.24–1.16(m,2H),0.86–0.78(m,2H),-0.10(s,9H).
MS(ESI)m/z 712.39(M+H)+.
(2)N-(4-(5-(1-(丁-2-炔酰基)哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶)-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)-3-甲基苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)哌啶-1-羧酸盐(216mg,0.303mmol)加入瓶中,加入2.5mL无水乙醇,室温搅拌均匀,滴加浓盐酸(1.2mL,15.15mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于3mL二氯甲烷,在冰盐浴的条件下加入DIEA(160μL,0.909mmol)和NaOH(17mg,0.42mmol),滴加2-丁炔酰氯(50μL,0.5454mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体125mg,收率60.85%。
熔点74-75℃
1H NMR(500MHz,DMSO-d6)δ8.98–8.87(m,1H),8.83(s,1H),7.83(d,J=8.1Hz,2H),7.56(s,1H),7.47(d,J=8.1Hz,2H),7.42(d,J=6.3Hz,3H),5.60(s,2H),4.55(d,J=5.8Hz,2H),4.22(d,J=13.1Hz,1H),4.11(d,J=13.6Hz,1H),3.54(t,J=7.9Hz,2H),2.67(q,J=14.0,12.8Hz,2H),2.46(s,3H),2.23(t,J=12.2Hz,1H),1.97(s,3H),1.54(s,2H),1.30(s,9H),1.23–1.13(m,2H),0.82(t,J=7.9Hz,2H),-0.09(s,9H).
MS(ESI)m/z 678.38(M+H)+.
(3)标题化合物的制备
将N-(4-(5-(1-(丁-2-炔酰基)哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶)-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺(68mg,0.1mmol)溶于1.5mL二氯甲烷,在冰浴条件下滴加三氟乙酸(670μL,9mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体35mg,收率63.90%,熔点162-163℃。
1H NMR(500MHz,DMSO-d6)δ12.01(s,1H),8.94–8.89(m,1H),8.74(s,1H),7.83(d,J=7.9Hz,2H),7.47(d,J=7.7Hz,2H),7.41(d,J=4.4Hz,3H),7.36(s,1H),4.55(d,J=4.3Hz,2H),4.22(d,J=12.9Hz,1H),4.11(d,J=17.0Hz,1H),2.65(t,J=11.9Hz,2H),2.46(s,3H),2.21(t,J=12.5Hz,1H),1.97(s,3H),1.60–1.49(m,2H),1.30(s,9H),1.24(s,2H).
HRMS calcd.For C34H38O2N5(M+H)+548.3020,found 548.3013.
实施例16
N-(4-(5-(1-丙烯酰基哌啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺
(1)N-(4-(5-(1-丙烯酰哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺的制备
将叔丁基4-(4-(4-((4-(叔丁基)苯甲酰胺基)甲基)-3-甲基苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)哌啶-1-羧酸盐(216mg,0.303mmol)加入瓶中,加入2.5mL无水乙醇,室温搅拌均匀,滴加浓盐酸(1.2mL,15.15mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于3mL二氯甲烷,在冰盐浴的条件下加入DIEA(160μL,0.909mmol)和NaOH(17mg,0.42mmol),滴加丙烯酰氯(55μL,0.682mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体103mg,收率51.04%,熔点73-74℃。
1H NMR(400MHz,DMSO-d6)δ8.89(t,J=5.8Hz,1H),8.78(s,1H),7.81–7.74(m,2H),7.50(d,J=1.0Hz,1H),7.44–7.34(m,5H),6.61(dd,J=16.7,10.4Hz,1H),5.98(dd,J=16.7,2.5Hz,1H),5.58–5.49(m,3H),4.50(d,J=5.8Hz,2H),4.32(d,J=12.8Hz,1H),3.82(d,J=13.6Hz,1H),3.54–3.42(m,2H),2.61(dt,J=53.7,12.2Hz,2H),2.41(s,3H),2.23–2.12(m,1H),1.47(dd,J=45.5,13.2Hz,2H),1.24(s,9H),1.22–1.17(m,2H),0.83–0.70(m,2H),-0.15(s,9H).
MS(ESI)m/z 666.37(M+H)+.
(2)标题化合物的制备
将N-(4-(5-(1-丙烯酰哌啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-甲基苄基)-4-(叔丁基)苯甲酰胺(67mg,0.1mmol)溶于1.5mL二氯甲烷,在冰浴条件下滴加三氟乙酸(670μL,9mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体27mg,收率50.40%,熔点164-165℃。
1H NMR(500MHz,DMSO-d6)δ12.01(s,1H),8.93(s,1H),8.74(s,1H),7.83(d,J=6.6Hz,2H),7.53–7.37(m,6H),7.34(s,1H),6.70–6.59(m,1H),6.02(d,J=17.0Hz,1H),5.59(d,J=9.0Hz,1H),4.55(s,2H),4.37(d,J=11.4Hz,1H),3.86(d,J=11.2Hz,1H),2.73–2.64(m,1H),2.64–2.54(m,1H),2.46(s,3H),2.22(s,1H),1.57(d,J=11.9Hz,1H),1.46(d,J=7.9Hz,1H),1.29(s,9H),1.24(s,2H).
HRMS calcd.For C33H38O2N5(M+H)+536.3020,found 536.3013.
实施例17
1-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶)-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-3-苯基脲
(1)(4-溴-2-氟苄基)氨基甲酸苯酯的制备
将4-溴-2-氟苄胺(700mg,3.362mmol)溶于4.5mL DMF,在冰盐浴条件下滴加吡啶(540μL,6.724mmol),滴加氯甲酸苯酯(645μL,5.043mmol),室温下搅拌10h。加水,析出白色固体,减压抽滤,收集滤饼,得白色固体1g,收率91.83%,熔点77-78℃。
1H NMR(500MHz,DMSO-d6)δ8.31(t,J=5.8Hz,1H),7.54(d,J=9.8Hz,1H),7.51–7.42(m,2H),7.37(t,J=7.5Hz,3H),7.20(t,J=7.6Hz,1H),7.11(d,J=7.9Hz,1H),4.28(d,J=6.0Hz,2H).
MS(ESI)m/z 324.00(M+H)+.
(2)1-(4-溴-2-氟苄基)-3-苯基脲的制备
将(4-溴-2-氟苄基)氨基甲酸苯酯(1.383g,4.267mmol)溶于DMF,加入二异丙基乙胺(3mL,17.068mmol),滴加苯胺(780μL,8.534mmol),室温下搅拌5h。加水,析出白色固体,减压抽滤,收集滤饼,得白色固体1.14g,收率82.67%。
熔点98-99℃
1H NMR(500MHz,DMSO-d6)δ8.59(s,1H),7.54–7.46(m,1H),7.45–7.30(m,4H),7.21(t,J=7.8Hz,2H),6.89(t,J=7.4Hz,1H),6.63(t,J=6.1Hz,1H),4.30(d,J=5.9Hz,2H).
MS(ESI)m/z 323.02(M+H)+.
(3)1-(2-氟-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)-3-苯基脲的制备
将1-(4-溴-2-氟苄基)-3-苯基脲(500mg,1.548mmol),联硼酸频那醇酯(590mg,2.322mmol)和醋酸钾(456mg,4.644mmol)加入瓶中,加入8mL二氧六环,室温搅拌均匀,加入Pd(dppf)Cl2(113mg,0.1548mmol),氩气保护下加热至100℃反应6h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=5:1,得白色固体468mg,收率81.68%,熔点117-118℃。
1H NMR(400MHz,DMSO-d6)δ8.61(s,1H),7.50–7.44(m,1H),7.42–7.36(m,3H),7.32(dd,J=10.5,1.0Hz,1H),7.24–7.17(m,2H),6.89(tt,J=7.3,1.2Hz,1H),6.63(t,J=6.1Hz,1H),4.36(d,J=6.0Hz,2H),1.29(s,12H).
MS(ESI)m/z 371.19(M+H)+.
(4)叔丁基4-(4-(3-氟-4-((3-苯基脲基)甲基)苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐的制备
将4-(4-氯)叔丁基-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(358mg,0.77mmol),1-(2-氟-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苄基)-3-苯基脲(368mg,1mmol)和碳酸钾(213mg,1.54mmol)加入瓶中,加入4mL二氧六环和2mL水,室温搅拌均匀,加入Pd(dppf)Cl2(56mg,0.077mmol),氩气保护下加热至100℃反应7h。将反应冷却至室温,加水,减压抽滤,收集滤液,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=2:1,得浅黄色固体428mg,收率82.63%,熔点85-86℃。
1H NMR(400MHz,DMSO-d6)δ8.91(s,1H),8.61(s,1H),7.78(s,1H),7.50(d,J=4.2Hz,2H),7.45–7.35(m,3H),7.21(ddd,J=9.8,5.8,2.3Hz,2H),6.95–6.84(m,1H),6.69(t,J=6.0Hz,1H),5.66(s,2H),5.45–5.18(m,1H),4.43(d,J=5.9Hz,2H),3.74(s,2H),3.57(dd,J=8.5,7.4Hz,2H),3.23(t,J=5.5Hz,2H),1.95(s,2H),1.39(s,9H),0.91–0.78(m,2H),-0.08(s,9H).
MS(ESI)m/z 673.34(M+H)+.
(5)1-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-3-苯基脲的制备
将叔丁基4-(4-(3-氟-4-((3-苯基脲基)甲基)苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(204mg,0.303mmol)加入瓶中,加入2.5mL无水乙醇,室温搅拌均匀,滴加浓盐酸(1.2mL,15.15mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于3mL二氯甲烷,在冰盐浴的条件下加入DIEA(160μL,0.909mmol)和NaOH(17mg,0.42mmol),滴加2-丁炔酰氯(50μL,0.5454mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体98mg,收率50.63%,熔点151-152℃。
1H NMR(500MHz,DMSO-d6)δ8.91(s,1H),8.61(s,1H),7.79(d,J=20.3Hz,1H),7.46(d,J=7.1Hz,2H),7.41–7.37(m,2H),7.20(t,J=7.8Hz,2H),6.89(t,J=7.4Hz,1H),6.75–6.59(m,2H),6.18–6.02(m,1H),5.66(s,2H),4.41(d,J=5.8Hz,2H),3.92(s,2H),3.58(t,J=8.1Hz,2H),3.46(d,J=25.4Hz,2H),2.01(d,J=31.9Hz,2H),1.24(s,3H),0.86–0.83(m,2H),-0.08(s,9H).
MS(ESI)m/z 639.34(M+H)+.
(6)标题化合物的制备
将1-(4-(5-(1-(丁-2-炔酰基)-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-3-苯基脲(64mg,0.1mmol)溶于1.5mL二氯甲烷,在冰浴条件下滴加三氟乙酸(670μL,9mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体31mg,收率61.02%,熔点156-157℃。
1H NMR(400MHz,DMSO-d6)δ12.38(s,1H),9.00(d,J=9.6Hz,1H),8.83(s,1H),7.60(dd,J=13.9,2.4Hz,1H),7.47(dt,J=14.7,8.0Hz,2H),7.40(d,J=7.6Hz,3H),7.20(t,J=7.9Hz,2H),7.00–6.93(m,1H),6.88(t,J=7.3Hz,1H),5.30(d,J=28.5Hz,1H),4.42(d,J=5.5Hz,2H),4.03(dd,J=14.2,7.1Hz,1H),3.85(s,1H),3.60–3.54(m,1H),3.46–3.38(m,1H),1.98(d,J=7.2Hz,5H).
HRMS calcd.For C29H26O2N6F(M+H)+509.2096,found 509.2087.
实施例18
1-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7H-吡咯并[2,3-d]嘧啶-4-基)-2-氟苄基)-3-苯基脲
(1)1-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)-2-氟苄基)-3-苯基脲的制备
将叔丁基4-(4-(3-氟-4-((3-苯基脲基)甲基)苯基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-5-基)-3,6-二氢吡啶-1(2H)-羧酸盐(204mg,0.303mmol)加入瓶中,加入2.5mL无水乙醇,室温搅拌均匀,滴加浓盐酸(1.2mL,15.15mmol),室温搅拌10h。冰浴条件下加入饱和碳酸氢钠溶液调pH至7~8,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥,浓缩得油状物。将油状物溶于3mL二氯甲烷,在冰盐浴的条件下加入DIEA(160μL,0.909mmol)和NaOH(17mg,0.42mmol),滴加丙烯酰氯(55μL,0.682mmol),室温搅拌3h。加水,二氯甲烷萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,柱层析分离,洗脱剂为石油醚:乙酸乙酯=1:1,得白色固体93mg,收率48.97%,熔点67-68℃。
1H NMR(500MHz,DMSO-d6)δ8.91(s,1H),8.61(s,1H),7.79(d,J=20.3Hz,1H),7.42(dd,J=39.5,7.4Hz,5H),7.20(t,J=7.7Hz,2H),6.89(t,J=7.4Hz,1H),6.69(dt,J=18.5,7.9Hz,2H),6.12(td,J=10.9,9.8,6.0Hz,1H),5.65(d,J=10.6Hz,3H),5.32(d,J=100.4Hz,1H),4.41(d,J=5.8Hz,2H),3.92(s,2H),3.58(t,J=8.2Hz,2H),3.46(d,J=25.4Hz,2H),2.01(d,J=31.9Hz,2H),0.85(t,J=7.7Hz,2H),-0.08(s,9H).
MS(ESI)m/z 627.35(M+H)+.
(2)标题化合物的制备
将1-(4-(5-(1-丙烯酰基-1,2,3,6-四氢吡啶-4-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-)d]嘧啶-4-基)-2-氟苄基)-3-苯基脲(63mg,0.1mmol)溶于1.5mL二氯甲烷,在冰浴条件下滴加三氟乙酸(670μL,9mmol),室温搅拌7h。在冰浴条件下加入NaOH水溶液调pH至7~8,室温搅拌1.5h。加水,乙酸乙酯萃取三次,饱和氯化钠溶液洗1次,无水硫酸钠干燥。浓缩,得白色固体25mg,收率50.40%,熔点187-188℃。
1H NMR(400MHz,DMSO-d6)δ12.30(d,J=10.0Hz,1H),8.78(s,1H),8.63(s,1H),7.61–7.51(m,1H),7.37(dd,J=27.4,7.0Hz,5H),7.15(t,J=7.8Hz,2H),6.84(t,J=7.3Hz,1H),6.64(dd,J=17.0,5.6Hz,2H),6.06(dd,J=17.7,6.0Hz,1H),5.60(dd,J=23.2,11.4Hz,1H),5.21(d,J=92.2Hz,1H),4.36(d,J=5.7Hz,2H),3.86(s,2H),3.47–3.33(m,2H),2.01(s,1H),1.92(s,1H).
HRMS calcd.For C28H26O2N6F(M+H)+497.2096,found 497.2111.
药理实验
实验例1、对BTK的抑制活性
实验目的:对本发明化合物进行体外BTK的抑制活性评价。
实验方法:实验使用来自Promega Corporation的ADP-GloTM检测试剂盒。共设置了11个浓度(10μM至1nM)。将50μL化合物添加到384孔稀释板中,并使用Echo将每行0.05μL稀释的化合物溶液转移到384测定板中,每列包含2个重复。将2.5μL酶工作液加入384孔检测板中,与化合物在25℃孵育15分钟。加入2.5μL底物工作液引发反应,60min后加入4μL ADPGlo试剂。60min后,加入8μL激酶检测试剂,25℃孵育60min。用envision Perkin Elmer读板器(Envision,PE,USA)读取发光信号。
实验结果:如表1-1所示,本发明化合物具有较高的BTK抑制活性,绝大多数半数抑制浓度IC50小于10nM。如表1-2所示,多个化合物的活性与阳性对照依鲁替尼相当。
表1-1本发明化合物对BTK蛋白的抑制活性范围
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注:抑制活性分级,“A”表示IC50值小于10nM,“B”表示IC50值在10nM与100nM之间,“C”表示IC50值在100nM与1000nM之间,“D”表示IC50值大于1000nM。
表1-2本发明化合物对BTK蛋白的抑制活性数值
实验例2、对TMD-8细胞的增殖抑制活性
实验目的:本实验通过检测化合物在TMD8细胞系中对体外细胞活性的影响而研究化合物抑制细胞增殖的作用。Ibrutinib为内控化合物。
实验方法:实验采用了96孔板的体外筛选体系,在每孔中加入95μL细胞悬液,在Min对照孔中加入不含细胞(含0.1%DMSO)的培养液。取5μL的20×化合物工作液加入到细胞培养板中。在Max对照中加入5μL DMSO细胞培养液混合液。DMSO终浓度为0.1%。将培养板在37℃,5%CO2培养箱中培养72小时。用CellTiter-Glo发光法进行细胞活性检测。SpectraMax Paradigm读数得出对应的每孔荧光值RLU。细胞增殖抑制率(InhibitionRate)数据采用下列公式来处理:Inhibition Rate(Inh%)=100-(RLUDrug-RLUMin)/(RLUMax-RLUMin)*100%。在EXCEL中计算不同浓度化合物对应的抑制率,然后用GraphPadPrism软件作抑制率曲线图并计算相关参数,参数包括细胞最大和最小抑制率,IC50值。
实验结果:如表2-1所示,本发明化合物具有较高的TMD-8增殖抑制活性,绝大多数半数抑制浓度IC50小于10nM。如表2-2所示,多个化合物的活性优于阳性对照依鲁替尼。
表2-1本发明化合物对TMD-8增殖抑制活性范围
注:抑制活性分级,“A”表示IC50值小于10nM,“B”表示IC50值在10nM与100nM之间,“C”表示IC50值在100nM与1000nM之间,“D”表示IC50值大于1000nM。
表2-2本发明化合物对TMD-8增殖抑制活性数值
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实验例3、体内药代动力学研究
实验目的:测试实施例化合物的体内药代动力学参数
实验方法:选用雄性BALB/c小鼠,分别为口服给药组和静脉注射组。对于口服给药组,10mg/kg剂量口服给药,分别于5min,10min,15min,30min,1h,2h,4h,6h,8h,12h于眼眶静脉丛交叉取血。对于静脉注射组,2mg/kg剂量静脉注射给药,分别于2min,5min,10min,15min,30min,1h,2h,4h,6h,8h,12h于眼眶静脉丛交叉取血。用离心法制备血浆,建立标准曲线,加入乙腈沉淀血浆蛋白后,用液相色谱-质谱联用(LC-MS/MS)分析上清液。绘制药时曲线,用WinNonLin软件分析数据,经非房室模型拟合,得到血浆药代动力学参数。
实验结果:如表3所示,口服剂量为10mg/kg时,实施例8化合物的口服生物利用度为40.98%,口服剂量为25mg/kg时,实施例8的口服生物利用度达到96.56%,而阳性药依鲁替尼的口服生物利用度只有4%。与阳性药相比,实施例8改善了BTK不可逆抑制剂生物利用度较低的问题。
表3口服给药和静脉注射给药实施例8后小鼠的药代动力学参数a
a药代动力学参数为3次实验测定结果的平均值。
实验例4、体内药效学研究
实验目的:测试实施例化合物8的体内抗肿瘤活性
实验方法:选用雌性CB17SCID小鼠,皮下接种TMD8细胞,在肿瘤平均体积达到约150-200mm3时开始分组给药。以依鲁替尼为阳性对照药,按照25mg/kg的剂量口服给药;对于本发明的化合物,分别按照12.5mg/kg、25mg/kg和50mg/kg的剂量口服给药。连续给药21天,每天两次。
实验结果:如图1所示,实施例化合物8低中高三个剂量组对肿瘤均有抑制作用,且高剂量组对肿瘤的抑制作用和阳性药依鲁替尼相当。本发明中的化合物8展现了较强的体内抗肿瘤活性,为弥漫大B淋巴瘤等的治疗提供了新的选择。
Claims (10)
1.如通式(I)所示的化合物或其药学上可接受的盐:
其中,R选自且R3选自丙烯酰基,2-丁炔酰基;
R1,R2取代基为一个或多个,选自H,C1-8直链或支链烷基,C3-8环烷基,C1-8烷氧基,C1-8烷基氨基,卤素,羟基,氨基,氰基,且所述的C1-8直链或支链烷基进一步被一个或多个卤素,羟基,氨基或氰基取代。
2.根据权利要求1所述的化合物或其药学上可接受的盐,其特征在于,R1,R2选自H,甲基,乙基,正丙基,异丙基,正丁基,异丁基,叔丁基,正戊基,异戊基,环丙基,环丁基,环戊基,甲氧基,乙氧基,丙氧基,甲基氨基,乙基氨基,F,Cl,Br,I,羟基,氨基,氰基,或者选自被一个或多个卤素取代的甲基,乙基,正丙基,异丙基,正丁基,异丁基,叔丁基,正戊基,异戊基。
3.根据权利要求1所述的化合物或其药学上可接受的盐,其特征在于,R1选自H,甲基,乙基,三氟甲基,F,Cl,Br,或I;R2选自H,异丙基,异丁基,叔丁基,异戊基,环丙基,环丁基,环戊基。
4.根据权利要求1所述的化合物或其药学上可接受的盐,其特征在于,所述的化合物结构为:
5.权利要求1-4任一项的化合物或其药学上可接受的盐的制备方法,其特征在于,步骤如下:
路线(1):
试剂与条件:(a)(2-(氯甲氧基)乙基)三甲基硅烷,碳酸钾,N,N-二甲基甲酰胺,室温;(b)N-Boc-1,2,5,6-四氢吡啶-4-硼酸频哪醇酯,1,1'-二(二苯膦基)二茂铁二氯化钯,碳酸钾,1,4-二氧六环,100℃;(c)R1,R2取代苯基硼酸频哪醇酯,1,1'-二(二苯膦基)二茂铁二氯化钯,碳酸钾,1,4-二氧六环,100℃;(d)盐酸的乙醇溶液,室温;丙烯酰氯,N,N-二异丙基乙胺,二氯甲烷,室温;(e)三氟乙酸,二氯甲烷,室温;四氢呋喃溶液,氢氧化钠溶液,室温;
路线(2):
试剂与条件:(a)(2-(氯甲氧基)乙基)三甲基硅烷,碳酸钾,N,N-二甲基甲酰胺,室温;(b)N-Boc-1,2,5,6-四氢吡啶-4-硼酸频哪醇酯,1,1'-二(二苯膦基)二茂铁二氯化钯,碳酸钾,1,4-二氧六环,100℃;(c)R1,R2取代苯基硼酸频哪醇酯,1,1'-二(二苯膦基)二茂铁二氯化钯,碳酸钾,1,4-二氧六环,100℃;(d)氢气,钯碳,乙酸乙酯溶液,甲醇溶液,室温(e)盐酸的乙醇溶液,室温;丙烯酰氯,N,N-二异丙基乙胺,二氯甲烷,室温;(f)三氟乙酸,二氯甲烷,室温;四氢呋喃溶液,氢氧化钠溶液,室温;
其中R1、R2的定义同权利要求1-4任一项所述。
6.一种药物组合物,其特征在于,所述药物组合物中包含权利要求1-4中任一项所述的化合物或其药学上可接受的盐以及药学上可接受的载体。
7.权利要求1-4中任一项所述的化合物或其药学上可接受的盐在制备治疗与BTK功能相关的疾病的药物中的应用。
8.权利要求1-4任一项所述的化合物或其药学上可接受的盐在制备治疗类风湿性关节炎、系统性红斑狼疮的药物中的应用。
9.权利要求1-4任一项所述的化合物或其药学上可接受的盐在制备治疗非霍奇金性淋巴瘤药物中的应用。
10.权利要求1-4任一项所述的化合物或其药学上可接受的盐在制备治疗华式巨球蛋白血症、慢性淋巴细胞白血病或原发性中枢神经系统淋巴瘤药物中的应用。
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