CN116283664A - 一种硝基儿茶酚衍生物、包含其的药物组合物及其应用 - Google Patents

一种硝基儿茶酚衍生物、包含其的药物组合物及其应用 Download PDF

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CN116283664A
CN116283664A CN202310221983.0A CN202310221983A CN116283664A CN 116283664 A CN116283664 A CN 116283664A CN 202310221983 A CN202310221983 A CN 202310221983A CN 116283664 A CN116283664 A CN 116283664A
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蒋学阳
冯锋
邹满星
谢焕芳
柳文媛
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China Pharmaceutical University
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Abstract

本发明提供一种硝基儿茶酚衍生物、包含其的药物组合物及其应用，硝基儿茶酚衍生物包括如式(Ⅰ)或式(Ⅱ)所示的化合物、其药学上可接受的盐或其溶剂化物，它具有COMT/MAO‑B双靶点抑制活性，对于治疗PD也具有重要的应用价值。

Description

一种硝基儿茶酚衍生物、包含其的药物组合物及其应用
技术领域
本发明属于药物化学领域，具体涉及一种硝基儿茶酚衍生物、包含其的药物组合物及其应用
背景技术
帕金森病(Parkinson’s disease,PD)是一种严重的进行性神经退行性运动障碍，目前其致病原因尚不明确。黑质纹状体多巴胺神经元的退化，导致DA缺乏，是PD运动障碍的原因，其主要症状通常是运动不能、僵硬和震颤。目前治疗帕金森病的主要策略是提高大脑中的多巴胺(Dopamine,DA)水平，临床药物治疗以前体药物左旋多巴(L-Dopamine)为主，此外还包括DA受体激动剂、儿茶酚-O-甲基转移酶(Catechol-O-methyltransferase，COMT)抑制剂、B型单胺氧化酶(Monoamine Oxidase B,MAO-B)抑制剂、抗胆碱能药等。
COMT是一种负责代谢内源性神经递质的酶，通过镁离子催化，将甲基从其辅酶S-腺苷-L-蛋氨酸(S-Adenosyl-L-methionine,SAM)转移到内源性神经递质的一个羟基上，从而终止其生物活性。目前大多数已知的COMT抑制剂都是以硝基邻苯二酚为基础的化合物，例如上市药物托卡朋(Tolcapone)和恩他卡朋(Entacapone)，它们可通过抑制外周和中枢组织中COMT酶的活性，阻断左旋多巴的外周降解，并增加其血浆半衰期，从而使更多口服左旋多巴能够穿过血脑屏障(blood-brain barrier,BBB)进入大脑，因此被用作辅助左旋多巴治疗帕金森病。单胺氧化酶(Monoamine Oxidase)是一种线粒体酶，在外周和中枢神经组织中对生物胺的体内失活起着重要作用。在中枢神经(Central nervous system,CNS)中，多巴胺主要由MAO-B代谢。目前已有雷沙吉兰(Rasagiline)、司来吉兰(Selegiline)、沙芬酰胺(Safinamide)等MAO-B抑制剂被开发用于临床，经常单独或联合左旋多巴用于早期PD的治疗。此外过氧化氢(H2O2)是MAO-B代谢多巴胺过程中的副产物，可能使多巴胺能神经元受到氧化损伤。
COMT/MAO-B双靶点抑制剂理论上，可通过多途径、有效地抑制外周组织和中枢组织对左旋多巴和多巴胺的代谢，从而延长左旋多巴的半衰期及抑制DA代谢，从增加外源性DA和减少内源性DA代谢两个途径提高并维持体内DA含量。此外作为MAO-B抑制剂还可通过减少多巴胺代谢中生成的过氧化氢(H2O2)，从而避免多巴胺能神经元受到氧化损伤。因此开发COMT/MAO-B双靶点抑制剂，对于治疗PD具有合理性，也具有重要的研究与应用价值，目前尚未有COMT/MAO-B双靶点抑制剂出现。
发明内容
本发明的目的在于提供一种具有COMT/MAO-B酶抑制活性的硝基儿茶酚衍生物，以及包含其的药物组合物，本发明的另一目的在于提供所述的硝基儿茶酚衍生物的制备方法及应用。
为了实现上述目的，本发明所采用的技术方案为：
一种硝基儿茶酚衍生物,它包括如式(Ⅰ)或式(Ⅱ)所示的化合物、其药学上可接受的盐或其溶剂化物，式中，R1选自取代或未取代的酰胺、取代或未取代的C1-6烷基、取代或未取代的C1-6烷氧基、取代或未取代的胺基、取代或未取代的苯基、酯基、氰基、硝基或卤素，所述取代为单取代或多取代，所述取代基为C1-6烷基、卤素、氰基、苯基或硝基；
Figure BDA0004117058610000021
R2选自H、CN、硝基或卤素。
作为技术方案的进一步改进，R1选自-OCH3、
Figure BDA0004117058610000022
Figure BDA0004117058610000023
其中Y为/>
Figure BDA0004117058610000024
Z为/>
Figure BDA0004117058610000025
R2选自H或CN。
作为技术方案的进一步改进，它选自以下任一种化合物：
Figure BDA0004117058610000031
一种所述硝基儿茶酚衍生物的制备方法，包括以下步骤：
Figure BDA0004117058610000032
(1)将化合物1和活性亚甲基发生Knoevenagel缩合反应，得到化合物2；将化合物1用NaClO2发生Pinnick氧化反应得到化合物3；
(2)将化合物2分别与胺、醇进行缩合反应，得到如式(Ⅰ)所示的化合物；将化合物3分别与胺、醇进行缩合反应，得到如式(Ⅱ)所示的化合物。
本发明还公开一种药物组合物，它包含所述硝基儿茶酚衍生物及一种或多种药学上可接受的辅料。药学上可接受的辅料系指生产药品和调配处方时使用的赋形剂和附加剂，包括溶剂、抛射剂、增溶剂、助溶剂、乳化剂、着色剂、黏合剂、崩解剂、填充剂、润滑剂、润湿剂、渗透压调节剂、稳定剂、助流剂、矫味剂、防腐剂、助悬剂、包衣材料、芳香剂、抗黏合剂、整合剂、渗透促进剂、pH值调节剂、缓冲剂、增塑剂、表面活性剂、发泡剂、消泡剂、增稠剂、包合剂、保湿剂、吸收剂、稀释剂、絮凝剂与反絮凝剂、助滤剂、释放阻滞剂等，优选口服药用辅料如崩解剂、助溶剂、溶剂、润滑剂等。
作为技术方案的进一步改进，所述药物组合物的剂型为胶囊剂、粉剂、丸剂、片剂、颗粒剂、悬浮剂或注射剂。
因本发明的硝基儿茶酚衍生物同时具有COMT和MAO抑制活性，对帕金森病具有应用前景，本发明还公开一种所述硝基儿茶酚衍生物在制备预防和/或治疗帕金森病药物中的应用。
因本发明的硝基儿茶酚衍生物同时具有COMT抑制活性，本发明还公开一种所述硝基儿茶酚衍生物在制备COMT抑制剂中的应用。
因本发明的硝基儿茶酚衍生物同时具有MAO抑制活性，本发明还公开一种所述硝基儿茶酚衍生物在制备MAO抑制剂中的应用。
本发明相对现有技术具有突出的实质性特点和显著的进步，具体的说，本发明的的双靶点化合物，具有COMT及MAO抑制活性。进一步说，本发明的化合物具有较好BBB透过性，在制备预防或治疗帕金森综合征药物中具有重要的应用价值。再一步说，本发明的化合物制备方法简单，便于产业转化应用。
具体实施方式
下面通过具体实施方式，对本发明的技术方案做进一步的详细描述。
本公开中实施例中未注明具体条件的实验方法，通常按照常规条件，或按照原料或商品制造厂商所建议的条件。未注明具体来源的试剂，为市场购买的常规试剂。
化合物的结构通过核磁共振(NMR)确定，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。
实施例1：
(4-{[(3-氟苯基)甲基]氧基}苯基)甲烷胺(4a)
Figure BDA0004117058610000041
将化合物4-[[叔丁氧酰胺]甲基]-苯酚(2.23g，10mmol)溶于乙腈中，加入K2CO3(1.94g，15mmol)搅拌3min,加入3-氟溴苄(1.46ml，10mmol)，加热回流6h。反应液抽滤，滤液浓缩，溶于DCM(20ml)中，向其中滴加等体积TFA，室温条件下反应3h，反应液浓缩，得到化合物4a，产率为90％。1H NMR(300MHz,DMSO-d6，δppm):8.10(s,2H,-NH2),7.50(dd,J＝8.5,5.7Hz,2H,Ar-H),7.38(d,J＝8.6Hz,2H,Ar-H),7.23(t,J＝8.9Hz,2H,Ar-H),7.06(d,J＝8.6Hz,2H,Ar-H),5.12(s,2H,-OCH2-),3.96(s,2H,-CH2-).
实施例2：
(4-{[(4-氟苯基)甲基]氧基}苯基)甲烷胺(4b)
Figure BDA0004117058610000051
以4-氟溴苄替代3-氟溴苄，其他条件不变，重复实施例1的操作，得到化合物4b。1HNMR(300MHz,DMSO-d6，δppm):8.12(s,2H,-NH2),7.49–7.40(m,1H,Ar-H),7.38(d,J＝8.7Hz,2H,Ar-H),7.30(s,1H,Ar-H),7.27(dd,J＝5.3,3.3Hz,1H,Ar-H),7.20–7.12(m,1H,Ar-H),7.09–7.03(m,2H,Ar-H),5.17(s,2H,-OCH2-),3.96(s,2H,-CH2-).
实施例3：
4-(氨基甲基)-N-(丙-2-炔基)苯甲酰胺(5a)
Figure BDA0004117058610000052
将4-[(叔丁氧羰基氨基)甲基]苯甲酸(2.5g,10mmol)溶于DMF(6mL)，加入DIEA(1653μL,10mmol)，冰水浴中加入PyBOP(5.2g,10mmol)，搅拌0.5h，加入炔丙胺(768μL,12mmol)，室温反应6h，加入水30mL，析出白色固体，抽滤，滤饼烘干，溶于DCM(20ml)中，向其中滴加等体积TFA，室温条件下反应1h，反应液浓缩，用DCM重结晶得化合物5a，产率为80％。1HNMR(300MHz,DMSO-d6,δppm):9.02(t,J＝5.5Hz,1H,-CONH-),8.31(s,2H,-NH2),7.90(d,J＝8.3Hz,2H,Ar-H),7.55(d,J＝8.3Hz,2H,Ar-H),4.11(s,2H,-CH2-),4.07(dd,J＝5.5,2.5Hz,2H,-CONHCH2 -),3.16(t,J＝2.5Hz,1H,-CCH).
实施例4：
(2S)-2-({[4-(氨基甲基)苯基]羰基}氨基)丙酰胺(5b)
Figure BDA0004117058610000053
以L-丙氨酰胺盐酸盐替代炔丙胺，其他条件不变，重复实施例3的操作，得到化合物5b。1H NMR(300MHz,DMSO-d6，δppm):8.49(d,J＝7.5Hz,1H,-CONH-),8.32(s,2H,-NH2),7.94(d,J＝8.2Hz,2H,Ar-H),7.54(d,J＝8.2Hz,2H,Ar-H),7.43(s,1H,-CONH2),7.03(s,1H,-CONH2),4.42(m,J＝7.2Hz,1H,-CHCH3),4.11(s,2H,CH2Ar),1.34(d,J＝7.2Hz,3H,-CHCH3 ).
实施例5：
(2E)-2-氰基-3-(3,4-二羟基-5-硝基苯基)-N-(丙-2-炔基)丙-2-烯酰胺(C1)
Figure BDA0004117058610000061
将2a(250mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入炔丙胺(77μL,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C1，产率为25％，m.p.200-202℃，HPLC(93％methanol in water with 0.1％ HCOOH):99.1％,tR＝3.130min.1H NMR(300MHz,DMSO-d6,δppm):10.93(br s,1H,OH),8.88(t,J＝5.5Hz,1H,-CONH-),8.09(s,1H,CHAr),7.97(d,J＝2.1Hz,1H,Ar-H),7.80(d,J＝2.2Hz,1H,Ar-H),3.99(dd,J＝2.5Hz,2H,-NHCH2 -),3.18(t,J＝2.5Hz,1H,-CCH).13C NMR(75MHz,DMSO-d6,δppm):161.52,149.83,148.66,146.46,137.68,122.19,120.17,118.23,116.68,104.02,81.05,73.76,29.57.HRMS(m/z)：[M-H]-calcd.for C13H9N3O5：286.0469；found,286.0495.
实施例6：
N-[(2E)-2-氰基-3-(3,4-二羟基-5-硝基苯基)-1-氧亚基丙-2-烯基]-L-丙氨酸甲酯(C2)
Figure BDA0004117058610000062
将2a(250mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入L-丙氨酸甲酯盐酸盐(167mg,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C2，产率为30％，m.p.162-164℃，HPLC(93％methanol in water with 0.1％ HCOOH):97.8％,tR＝3.206min.1H NMR(300MHz,DMSO-d6,δppm):10.98(br s,2H,2×OH),8.78(d,J＝6.9Hz,1H,-CONH-),8.09(s,1H,CHAr),7.96(d,J＝2.1Hz,1H,Ar-H),7.81(d,J＝2.1Hz,1H,Ar-H),4.41(m,1H,-CHCH3),3.66(s,3H,-OCH3),1.39(d,J＝7.3Hz,3H m,1H,-CHCH 3 ).13C NMR(75MHz,DMSO-d6,δppm):172.96,161.87,149.71,148.69,137.67,136.80,122.17,120.28,118.07,116.64,104.12,52.53,49.11,17.05.HRMS(m/z)：[M-H]-calcd.for C14H13N3O7：334.0681；found,334.0713.
实施例7：
((2S)-2-{[(2E)-2-氰基-3-(3,4-二羟基-5-硝基苯基)-1-氧亚基丙-2-烯基]氨基}丙酰胺(C3)
Figure BDA0004117058610000071
将2a(250mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h；将L-丙氨酰胺盐酸盐(249mg,2mmol)溶于DMF(2mL)，加入DIEA(348μL,2mmol)，室温搅拌0.5h后将其加入到12b的DMF溶液中，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用甲醇重结晶得产物C3，产率为15％，m.p.236-238℃，HPLC(93％methanol in water with 0.1％ HCOOH):99.4％,tR＝2.987min.1H NMR(300MHz,DMSO-d6,δppm):10.96(br s,2H,2×OH),8.30(d,J＝7.2Hz,1H,-CONH-),8.11(s,1H,CHAr),7.95(d,J＝2.1Hz,1H,Ar-H),7.81(d,J＝2.2Hz,1H,Ar-H),7.50(s,1H,-NH2),7.15(s,1H,-NH2),4.31(m,1H,-CHCH3),1.32(d,J＝7.1Hz,3H,-CHCH3 ).13C NMR(75MHz,DMSO-d6,δppm):174.11,161.26,149.17,148.59,146.02,137.68,122.51,119.99,118.23,116.86,104.80,49.56,18.56.HRMS(m/z)：[M-H]-calcd.for C13H12N4O6：319.0684；found,319.0713.
实施例8：
(2E)-3-(3,4-二羟基-5-硝基苯基)-N-(丙-2-炔基)丙-2-烯酰胺(C4)
Figure BDA0004117058610000072
将2b(225mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入炔丙胺(77μL,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C4，产率为24％，m.p.202-204℃，HPLC(93％methanol in water with 0.1％ HCOOH):85.7％,tR＝2.950min.1H NMR(300MHz,DMSO-d6,δppm):10.53(br s,2H,2×OH),8.53(t,J＝5.5Hz,1H,-CONH-),7.56(d,J＝1.9Hz,1H,Ar-H),7.36(d,J＝15.8Hz,1H,-CHCHCO-),7.26(d,J＝1.9Hz,1H,Ar-H),6.48(d,J＝15.8Hz,-CHCHCO-),3.99(dd,J＝5.4,2.4Hz,2H,-NHCH2 -),3.17(t,J＝2.4Hz,1H,-CCH).13C NMR(75MHz,DMSO-d6,δppm):164.98,148.47,143.73,138.31,137.79,125.72,121.10,117.07,115.58,81.49,73.70,28.47.HRMS(m/z)：[M-H]-calcd.for C12H10N2O5：261.0517；found,261.0541.
实施例9：
N-[(2E)-3-(3,4-二羟基-5-硝基苯基)-1-氧亚基丙-2-烯基]-L-丙氨酸甲酯(C5)
Figure BDA0004117058610000081
将2b(225mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入L-丙氨酸甲酯盐酸盐(167mg,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C5，产率为31％，m.p.184-186℃，HPLC(93％methanol in water with 0.1％ HCOOH):99.6％,tR＝3.008min.1H NMR(300MHz,DMSO-d6,δppm):10.57(br s,2H,2×OH),8.53(d,J＝7.0Hz,1H,-CONH-),7.57(d,J＝1.9Hz,1H,Ar-H),7.34(d,J＝15.8Hz,1H,-CHCHCO-),7.26(d,J＝2.0Hz,1H,Ar-H),6.54(d,J＝15.8Hz,1H,-CHCHCO-),4.39(m,1H,-CHCH3),3.64(s,3H,-OCH3 ),1.33(d,J＝7.3Hz,3H,-CHCH3 ).13C NMR(75MHz,DMSO-d6,δppm):173.63,165.09,148.50,143.79,138.35,137.76,125.71,121.06,117.06,115.57,52.39,48.21,17.54.HRMS(m/z)：[M-H]-calcd.for C13H14N2O7：309.0728；found,309.0758.
实施例10：
(2S)-2-{[(2E)-3-(3,4-二羟基-5-硝基苯基)-1-氧亚基丙-2-烯基]氨基}丙酰胺(C6)
Figure BDA0004117058610000091
将2b(225mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h；将L-丙氨酰胺盐酸盐(249mg,2mmol)溶于DMF(2mL)，加入DIEA(348μL,2mmol)，室温搅拌0.5h后将其加入到12a的DMF溶液中，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用甲醇重结晶得产物C6，产率为16％，m.p.220-222℃，HPLC(93％methanol in water with 0.1％ HCOOH):99.7％,tR＝2.800min.1H NMR(300MHz,DMSO-d6,δppm):10.53(br s,2H,2×OH),8.22(d,J＝7.7Hz,1H,-CONH-),7.55(d,J＝1.9Hz,1H,Ar-H),7.44(s,1H,-NH2),7.30(d,J＝15.8Hz,1H,-CHCHCO-),7.26(d,J＝1.9Hz,1H,Ar-H),7.03(s,1H,-NH2),6.63(d,J＝15.7Hz,1H,-CHCHCO-),4.35(m,1H,-CHCH3),1.25(d,J＝7.1Hz,3H,-CHCH3 ).13C NMR(75MHz,DMSO-d6,δppm):174.70,164.77,148.39,143.35,137.68(d,J＝18.5Hz),126.18,122.11,117.32,115.33,48.50,19.05.HRMS(m/z)：[M+H]+calcd.for C12H13N3O6：296.0877；found,296.0881.
实施例11：
3,4-二羟基-5-硝基-N-(丙-2-炔基)苯甲酰胺(C7)
Figure BDA0004117058610000092
将3(199mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入炔丙胺(77μL,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C7，产率为24％，m.p.158-160℃，HPLC(93％methanol in water with 0.1％ HCOOH):99.8％,tR＝2.892min.1H NMR(300MHz,DMSO-d6,δppm):10.68(br s,2H,2×OH),9.00(t,J＝5.4Hz,1H,-CONH-),7.95(d,J＝2.1Hz,1H,Ar-H),7.59(d,J＝2.1Hz,1H,Ar-H),4.02(dd,J＝5.4,2.5Hz,2H,-NHCH2 -),3.14(t,J＝2.5Hz,1H,-CCH).13C NMR(75MHz,DMSO-d6,δppm):164.52,147.95,144.84,137.10,124.32,118.68,114.60,81.63,73.38,29.04.HRMS(m/z)：[M-H]-calcd.for C10H8N2O5：235.0360；found,235.0382.
实施例12：
N-[(3,4-二羟基-5-硝基苯基)羰基]-L-丙氨酸甲酯(C8)
Figure BDA0004117058610000101
将3(199mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入L-丙氨酸甲酯盐酸盐(167mg,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C8，产率为28％，m.p.120-122℃，HPLC(93％methanol in water with 0.1％ HCOOH):99.8％,tR＝2.942min.1H NMR(300MHz,DMSO-d6,δppm):10.67(br s,2H,2×OH),8.87(d,J＝6.8Hz,1H,-CONH-),8.01(d,J＝1.7Hz,1H,Ar-H),7.59(d,J＝1.7Hz,1H,Ar-H),4.45(m,1H,-CHCH3),3.64(s,3H,-OCH3),1.39(d,J＝7.3Hz,3H,-CHCH3 ).13C NMR(75MHz,DMSO-d6,δppm):173.60,164.79,147.97,145.01,137.10,123.99,118.69,114.79,52.38,48.82,17.13.HRMS(m/z)：[M-H]-calcd.for C11H12N2O7：283.0572；found,283.0598.
实施例13：
(2S)-2-{[(3,4-二羟基-5-硝基苯基)羰基]氨基}丙酰胺(C9)
Figure BDA0004117058610000102
将3(199mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h；将L-丙氨酰胺盐酸盐(249mg,2mmol)溶于DMF(2mL)，加入DIEA(348μL,2mmol)，室温搅拌0.5h后将其加入到19的DMF溶液中，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用甲醇重结晶得产物C9，产率为16％，m.p.218-220℃，HPLC(93％methanol in water with 0.1％ HCOOH):99.6％,tR＝2.750min.1H NMR(300MHz,DMSO-d6,δppm):10.59(br s,2H,2×OH),8.53(d,J＝7.4Hz,1H,-CONH-),8.02(d,J＝2.1Hz,1H,Ar-H),7.59(d,J＝2.1Hz,1H,Ar-H),7.39(s,1H,-NH2),6.99(s,1H,-NH2),4.36(m,1H,-CHCH3),1.31(d,J＝7.2Hz,3H,-CHCH3 ).13C NMR(75MHz,DMSO-d6,δppm):174.90,164.56,147.79,144.74,137.09,124.58,118.78,114.88,49.36,18.34.HRMS(m/z)：[M-H]-calcd.forC10H11N3O6：268.0575；found,268.0600.
实施例14：
(2E)-2-氰基-3-(3,4-二羟基-5-硝基苯基)丙-2-烯酸甲酯(C10)
Figure BDA0004117058610000111
将2a(250mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于甲醇(2mL)中的PyBOP(520mg,1.0mmol)，室温反应6h。低压浓缩，加水析出黄色固体，抽滤，滤饼用DCM重结晶得产物C10，产率为70％，m.p.220-222℃，HPLC(93％methanol inwater with 0.1％ HCOOH):94.2％,tR＝3.217min.1H NMR(300MHz,DMSO-d6,δppm):10.92(br s,2H,2×OH),8.31(s,1H,CHAr),8.12(d,J＝2.1Hz,1H,Ar-H),7.90(d,J＝2.2Hz,1H,Ar-H),3.85(s,3H,-CH3).13C NMR(75MHz,DMSO-d6,δppm):163.01,153.89,148.61,147.09,137.79,121.90,121.64,118.34,116.18,100.36,53.72.HRMS(m/z)：[M-H]-calcd.forC11H8N2O6：263.0310；found,263.0334.
实施例15：
(2E)-2-氰基-3-(3,4-二羟基-5-硝基苯基)-N-[(4-{[(3-氟苯基)甲基]氧基}苯基)甲基]丙-2-烯酰胺(C11)
Figure BDA0004117058610000112
将2a(250mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入4a(278mg,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C11，产率为26％，m.p.206-208℃，HPLC(93％methanol in water with 0.1％ HCOOH):98.8％,tR＝3.980min.1H NMR(300MHz,DMSO-d6,δppm):8.63(t,J＝5.6Hz,1H,-CONH-),7.93(s,1H,CHAr),7.88(d,J＝1.6Hz,1H,Ar-H),7.88(d,J＝1.6Hz,1H,Ar-H),7.56(d,J＝1.6Hz,1H,Ar-H),7.48–7.37(m,1H,Ar-H),7.33–7.21(m,4H,Ar-H),7.15(t,J＝8.2Hz,1H,Ar-H),6.98(d,J＝8.4Hz,2H,Ar-H),5.12(s,2H,-OCH2-),4.32(d,J＝5.4Hz,2H,-CONHCH2 -).13C NMR(75MHz,DMSO-d6,δppm):162.64(d,J＝243.7Hz),162.61,157.50,151.51,150.36,140.62(d,J＝7.9Hz),135.50,135.47,132.16,130.93(d,J＝8.4Hz),129.28,125.18,123.88(d,J＝2.3Hz),118.26,115.43,115.07,114.99(d,J＝24.2Hz),114.82,114.57(d,J＝21.9Hz),105.06,68.72,42.95.HRMS(m/z)：[M-H]-calcd.for C24H18FN3O6：426.1107；found,462.1149.
实施例16：
(2E)-2-氰基-3-(3,4-二羟基-5-硝基苯基)-N-[(4-{[(4-氟苯基)甲基]氧基}苯基)甲基]丙-2-烯酰胺(C12)
Figure BDA0004117058610000121
将2a(250mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入4b(278mg,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C12，产率为23％，m.p.208-210℃，HPLC(93％methanol in water with 0.1％ HCOOH):95.4％,tR＝3.990min.1H NMR(300MHz,DMSO-d6,δppm):8.86(t,J＝5.5Hz,1H,-CONH-),8.09(s,1H,CHAr),8.01(d,J＝1.5Hz,1H,Ar-H),7.78(d,J＝1.5Hz,1H,Ar-H),7.57(dd,J＝8.0,5.9Hz,2H,Ar-H),7.37–7.25(m,4H,Ar-H),7.05(d,J＝8.4Hz,2H,Ar-H),5.18(s,2H,-OCH2-),4.41(d,J＝5.4Hz,2H,-CONHCH2 -).13CNMR(75MHz,DMSO-d6,δppm):162.17(d,J＝243.8Hz),161.97,157.68,150.90,149.83,136.72,133.81(d,J＝3.0Hz),131.79,130.30(d,J＝8.3Hz),129.31,122.14,119.54,117.46,115.83,115.69(d,J＝21.1Hz),115.20,115.08,101.92,68.87,43.05.HRMS(m/z)：[M+H]+calcd.for C24H18FN3O6：464.1252；found,464.1252.
实施例17：
(2E)-3-(3,4-二羟基-5-硝基苯基)-N-[(4-{[(3-氟苯基)甲基]氧基}苯基)甲基]丙-2-烯酰胺(C13)
Figure BDA0004117058610000131
将2b(225mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入4a(278mg,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C13，产率为26％，m.p.202-204℃，HPLC(93％methanol in water with 0.1％ HCOOH):98.4％,tR＝3.728min.1H NMR(300MHz,DMSO-d6,δppm):10.54(br s,2H,2×OH),8.53(t,J＝5.8Hz,1H,-CONH-),7.55(d,J＝1.9Hz,1H,Ar-H),7.47-7.40(m,1H,Ar-H),7.35(d,J＝15.7Hz,1H,-CHCHCO-),7.27(d,J＝8.7Hz,2H,Ar-H),7.26(d,J＝2.0Hz,1H,Ar-H),7.22(d,J＝8.7Hz,2H,Ar-H),7.15(td,J＝8.2,2.1Hz,1H,Ar-H),6.98(d,J＝8.7Hz,2H,Ar-H),6.53(d,J＝15.7Hz,1H,-CHCHCO-),5.13(s,2H,-OCH2-),4.32(d,J＝5.7Hz,2H,-CONHCH2 -).13C NMR(75MHz,DMSO-d6,δppm):165.17,162.64(d,J＝243.7Hz),157.51,148.45,143.51,140.58(d,J＝7.4Hz),137.78,137.76,132.16,130.88(d,J＝8.3Hz),129.25,126.03,123.82(d,J＝2.7Hz),121.89,117.28,115.23(d,J＝21.9Hz),115.14,114.80,114.54(d,J＝21.9Hz),68.71,42.26.HRMS(m/z)：[M-H]-calcd.for C23H19FN2O6：437.1154；found,437.1196.
实施例18：
4-({[(2E)-3-(3,4-二羟基-5-硝基苯基)-1-氧亚基丙-2-烯基]氨基}甲基)-N-(丙-2-炔基)苯甲酰胺(C14)
Figure BDA0004117058610000132
将2b(225mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入4b(278mg,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C14，产率为26％，m.p.204-206℃，HPLC(93％methanol in water with 0.1％ HCOOH):91.6％,tR＝3.670min.1H NMR(300MHz,DMSO-d6,δppm):10.59(br s,2H,2×OH),8.52(s,1H,-CONH-),7.56(s,1H,Ar-H),7.50(d,J＝5.9Hz,2H,Ar-H),7.35(d,J＝15.8Hz,1H,-CHCHCO-),7.25(s,1H,Ar-H),7.25–7.16(m,4H,Ar-H),6.97(d,J＝7.5Hz,2H,Ar-H),6.54(d,J＝15.3Hz,1H,-CHCHCO-),5.08(s,2H,-OCH2-),4.32(d,J＝4.0Hz,2H,-CONHCH2 -).13C NMR(75MHz,DMSO-d6,δppm):165.11(s),162.63(d,J＝237.00Hz),157.65,148.40,143.37,137.84,137.75,133.83(d,J＝2.9Hz),132.05,130.30(d,J＝8.3Hz),129.23,126.07,121.92,117.32,115.69(d,J＝21.1Hz),115.30,115.16,68.88,42.23.HRMS(m/z)：[M-H]-calcd.for C23H19FN2O6：437.1154；found,437.1196.
实施例19：
N-[(4-{[(3-氟苯基)甲基]氧基}苯基)甲基]-3,4-二羟基-5-硝基苯甲酰胺(C15)
Figure BDA0004117058610000141
将3(199mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入4a(278mg,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C15，产率为23％，m.p.180-182℃，HPLC(93％methanol in water with 0.1％ HCOOH):97.4％,tR＝3.574min.1H NMR(300MHz,DMSO-d6,δppm):10.61(br s,2H,2×OH),9.02(t,J＝5.8Hz,1H,-CONH-),7.97(d,J＝2.1Hz,1H,Ar-H),7.59(d,J＝2.0Hz,1H,Ar-H),7.43(td,J＝8.0,6.1Hz,1H,Ar-H),7.29–7.22(m,4H,Ar-H),7.15(td,J＝8.4,1.9Hz,1H,Ar-H),6.97(d,J＝8.6Hz,2H,Ar-H),5.12(s,2H,-OCH2-),4.37(d,J＝5.7Hz,2H,-CONHCH2 -).13C NMR(75MHz,DMSO-d6,δppm):164.68,164.64(d,J＝243.7Hz),157.47,148.06,145.12,140.59(d,J＝7.4Hz),136.98,132.41,130.88(d,J＝8.3Hz),129.17,124.65,123.82(d,J＝2.7Hz),118.56,115.08,114.68(d,J＝18.1Hz),114.54(d,J＝21.9Hz),68.73,42.64.HRMS(m/z)：[M-H]-calcd.for C21H17FN2O6：411.0998；found,411.1037.
实施例20：
N-[(4-{[(4-氟苯基)甲基]氧基}苯基)甲基]-3,4-二羟基-5-硝基苯甲酰胺(C16)
Figure BDA0004117058610000151
将3(199mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入4b(278mg,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C16，产率为24％，m.p.176-178℃，HPLC(93％methanol in water with 0.1％ HCOOH):96.0％,tR＝3.524min.1H NMR(300MHz,DMSO-d6,δppm):10.60(br s,2H,2×OH),9.02(t,J＝5.7Hz,1H,-CONH-),7.97(d,J＝1.8Hz,1H,Ar-H),7.60(d,J＝1.8Hz,1H,Ar-H),7.49(dd,J＝8.3,5.7Hz,2H,Ar-H),7.24-7.18(dd,J＝9.6,8.5Hz,4H,Ar-H),6.96(d,J＝8.5Hz,2H,Ar-H),5.07(s,2H,-OCH2-),4.37(d,J＝5.6Hz,2H,-CONHCH2 -).13C NMR(75MHz,DMSO-d6,δppm):164.62,162.16(d,J＝243.0Hz),157.59,147.99,144.94,137.04,133.83(d,J＝2.9Hz),132.30,130.30(d,J＝8.3Hz),129.14,124.70,118.56,115.69(d,J＝21.1Hz),115.08,114.49,68.87,42.62.HRMS(m/z)：[M-H]- calcd.for C21H17FN2O6：411.0998；found,411.1026.
实施例21：
4-({[(2E)-3-(3,4-二羟基-5-硝基苯基)-1-氧亚基丙-2-烯基]氨基}甲基)-N-(丙-2-炔基)苯甲酰胺(C17)
Figure BDA0004117058610000152
将2b(225mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入5a(226mg,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C17，产率为24％，m.p.242-244℃，HPLC(93％methanol in water with 0.1％ HCOOH):96.1％,tR＝2.840min.1H NMR(300MHz,DMSO-d6,δppm):10.56(br s,2H,2×OH),8.91(t,J＝5.5Hz,1H,-CHCHCONH-),8.67(t,J＝6.0Hz,1H,Ar-CONH),7.83(d,J＝8.3Hz,2H,Ar-H),7.58(d,J＝1.9Hz,1H,Ar-H),7.38(d,J＝15.7Hz,1H,-CHCHCONH-),7.38(d,J＝8.3Hz,2H,Ar-H),7.27(d,J＝2.0Hz,1H,Ar-H),6.56(d,J＝15.7Hz,1H,-CHCHCONH-),4.46(d,J＝5.8Hz,2H,CH2-Ar),4.05(dd,J＝5.5,2.4Hz,2H,-CH2 CCH),3.13(t,J＝2.5Hz,1H,-CH2CCH).13C NMR(75MHz,DMSO-d6,δppm):166.15,165.38,148.43,143.52,143.44,138.04,137.81,132.85,127.85,127.59,125.96,121.66,117.32,115.41,81.84,73.31,42.46,28.92.HRMS(m/z)：[M-H]-calcd.for C20H17N3O6：394.1045；found,394.1079.
实施例22：
(2S)-2-({[4-({[(2E)-3-(3,4-二羟基-5-硝基苯基)-1-氧亚基丙-2-烯基]氨基}甲基)苯基]羰基}氨基)丙酰胺(C18)
Figure BDA0004117058610000161
将2b(225mg,1.0mmol)溶于DMF(2mL)，加入DIEA(174μL,1mmol)，冰水浴中加入溶于DCM(2mL)中的PyBOP(520mg,1.0mmol)，搅拌0.5h，然后加入5b(266mg,1.2mmol)，室温反应6h。低压浓缩，加入乙酸乙酯(30mL)，依次用水(3×10mL)、HCl 1M(3×10mL)和盐水(10mL)洗涤，浓缩有机层，用DCM重结晶得产物C18，产率为23％，m.p.178-180℃，HPLC(93％methanol in water with 0.1％ HCOOH):99.3％,tR＝2.075min.1H NMR(300MHz,DMSO-d6,δppm):10.54(br s,2H,2×OH),8.67(t,J＝6.0Hz,1H,-CHCHCONH-),8.38(d,J＝7.5Hz,1H,Ar-CONH),7.87(d,J＝8.3Hz,2H,Ar-H),7.58(d,J＝1.9Hz,1H,Ar-H),7.38(d,J＝15.7Hz,1H,-CHCHCONH-),7.37(d,J＝8.5Hz,2H,Ar-H),7.38(s,1H,NH2),7.27(d,J＝2.0Hz,1H,Ar-H),7.01(s,1H,NH2),6.57(d,J＝15.7Hz,1H,-CHCHCONH-),4.46(d,J＝5.8Hz,2H,CH2-Ar),4.40(m,1H,-CHCH3),1.33(d,J＝7.2Hz,3H,-CHCH3 ).13C NMR(75MHz,DMSO-d6,δppm):174.93,166.17,165.36,148.42,143.47,143.19,138.01,137.83,133.20,128.06,127.39,125.98,121.69,117.33,115.39,49.17,42.48,18.53.HRMS(m/z)：[M-H]-calcd.forC20H20N4O7：427.1259；found,427.1296.
实施例23：体外(COMT)抑制活性测试
药品与试剂：实施例中制得的化合物、大鼠肝胞浆COMT酶(自制，SD大鼠，SPF级别，购自江苏华创信诺医药科技有限公司)、底物秦皮乙素(江苏艾康生物医药研发有限公司)、SAM(上海毕得医药科技股份有限公司)、二硫苏糖醇(上海贤鼎生物科技有限公司)。
仪器：多功能酶标仪(Synergy 2，BioTek Instruments,Inc)，高速冷冻微型离心机(D3024R型，Scilogex)，匀浆机(F6/10型，上海净信实业发展有限公司)。
大鼠肝胞浆COMT酶的制备：大鼠麻醉，腹主动脉取血后，取肝脏用冰冷盐水清洗,滤纸吸干水分，-20℃冷冻1h，冰冻肝组织，切成小片，在2倍体积PBS(50mM，pH 7.5，含有0.5mM二硫苏糖醇)中匀浆2分钟。匀浆在4℃下离心，10000×g离心15min，去除上层漂浮脂肪，上清液在4℃下离心，21000×g离心15min，上清为大鼠肝胞浆COMT酶，分装为2ml/支(每支供100孔分析)-80℃冷冻。
COMT酶活测试步骤：(1)向黑色96孔板加入PBS(25mM，pH 7.8，含有2mM MgCl2，20mM L-半胱氨酸，1mM SAM，20μM秦皮乙素)170μL和抑制剂10μL，预孵育15min，37℃；(2)加入大鼠肝胞浆COMT酶20μL启动反应，使反应终体积为200μL；(3)使连续记录荧光强度(λex＝360nm,λem＝485nm)500s；(4)构建荧光强度-时间曲线，取线性反应期曲线斜率，即单位时间内荧光强度的变化ΔInt/min作为报告单位；(5)抑制能力用剩余活性(Ra％)表示，公式为：Ra％＝(加抑制剂孔的ΔInt/min)/(不加抑制剂孔的ΔInt/min)。所得结果用GraphPadPrismTM软件以非线性衰退分析模式(non-liner regression analysis model)计算得相应IC50值，如表1所示。
实施例24：体外(MAO)抑制活性测试
药品与试剂：实施例中制得的化合物、大鼠脑MAO酶(自制，SD大鼠，SPF级别，购自江苏华创信诺医药科技有限公司)、MAO-B底物苄胺盐酸盐(上海毕得医药科技股份有限公司)、MAO-A底物苄5-羟色胺(上海吉至生化有限公司)、Amplex Red(上海贤鼎生物科技有限公司)、HRP(碧云天生物)、
仪器：多功能酶标仪(Synergy 2，BioTek Instruments,Inc)，高速冷冻微型离心机(D3024R型，Scilogex)，匀浆机(F6/10型，上海净信实业发展有限公司)
大鼠脑线粒体MAO酶的制备：大鼠处死后，取全脑去除嗅球，用冰冷盐水清洗，滤纸吸干水分，-20℃冷冻1h，在5倍体积PBS(100mM，pH 7.4，0.1％ TritonX-114)中匀浆2min。匀浆在4℃下离心，3000×g离心20min，上清为大鼠脑线粒体MAO酶，-80℃冷冻。
MAO酶活测试步骤：(1)在平底黑色96孔微孔板中，加入大鼠脑线粒体MAO酶90μL，加入用甲醇稀释的抑制剂10μL，在37℃的温度下，避光，预孵育10分钟；(2)预孵育后，加入MAO-B显色剂(AR/HRP/苄胺)或MAO-A显色剂(AR/HRP/5-HT)100μL，终体积为200μL，孵育15min；(3)测定resorufin产生的荧光(λex＝530nm,λem＝590nm)；(4)空白值(b)，用1XBuffer代替MAO酶；Vi是测试化合物存在时的速度，Vo是甲醇代替抑制剂存在时的控制速度；(5)抑制能力用剩余活性表示，公式为：Ra＝(Vi-b)/(Vo-b)；所得结果用GraphPad PrismTM软件以非线性衰退分析模式(non-liner regression analysis model)计算得相应IC50值，如表1和表2所示。
表1 化合物对rat liver COMT、rat brain MAO-A和rat brain MAO-B抑制活性
Figure BDA0004117058610000181
/>
Figure BDA0004117058610000191
结果分析：从活性测试总体结果上看，化合物C1-C18均具有较优的COMT抑制活性，在给予化合物10μM浓度下MAO-A的残余活性大于90％，MAO-B的残余活性在50％～90％，说明化合物具有一定的选择性抑制MAO-B能力，其中C11和C12对MAO-B的抑制能力最强，进一步进行了其对MAO-B的IC50值测定，如表2所示。
表2 优选化合物对rat liver COMT、rat brain MAO-B的IC50值
Figure BDA0004117058610000192
结果分析：化合物C11和C12对MAO-B的抑制能力接近，化合物C12相对C11具有更强的COMT抑制活性，
实施例25：实施例18口服给药后不同时间在血液以及脑中检测量(化合物C11)
雄性ICR小鼠8只，禁食12h，每只小鼠口服灌胃单剂量化合物20mg/kg(采用VDMSO：VPEG400：V生理盐水＝1：49：50配成浓度2mg/mL的溶液)，在30min、50min、80min、120min时从眼眶窦收集1mL血样在肝素化管中。血浆上清即通过离心分离，取上清用甲醇稀释至1:3，涡旋5min，4℃，15000rpm离心15min，待进样分析。同时取出脑组织样本，然后大脑在滤纸上滚动，称重(记录脑重)，放入-80℃冰箱15min，直接加1倍甲醇：生理盐水(3:7,v/v)稀释，将每个组织样品均质化，在4℃，以15000rpm离心15min。然后取上清加等体积甲醇沉淀蛋白，涡旋5min，4℃，15000rpm离心15min，取上清过滤膜，用LC-MS/MS进样分析。
LC-MS/MS条件如下：色谱柱为C18柱(150×4.6mm，5μM)；流动相为：A相为纯水，B相为有机相(甲醇)；采用等度洗脱：有机相比例为93％；流速：0.5mL/min，进样量：10.0μL，柱温：35℃。
结果见表。给药后30min在血中检测C11含量最高，到50min时迅速降为最低，随后在80min含量二次升高，可能是因为肝肠循环；在脑组织中检测到C11的含量在20～300ng·g-1范围内，在30～180min内C11脑内含量随时间增加先升高再下降，在120min时含量最高为298.47±176.41ng·g-1，此时C11在脑内浓度约为血清中浓度的两倍，说明C11具有良好的BBB透过性。在相同时间点120min，化合物C11较C12在脑组织浓度更高，C11可能具有更好的入脑效果。
表3 化合物C11和C12口服给药后不同时间在小鼠血清以及脑组织浓度
化合物 时间/min 血清浓度/ng·ml-1 脑组织浓度/ng·g-1
C11 30 1068.70±332.15 26.58±14.59
C11 50 85.68±42.11 48.21±24.54
C11 80 252.80±8.80 232.80±20.00
C11 120 159.32±140.59 298.47±176.41
C11 180 249.80±143.4 102.20±17.40
C12 120 46.78±13.60 97.40±3.68
最后应当说明的是：以上实施例仅用以说明本发明的技术方案而非对其限制；尽管参照较佳实施例对本发明进行了详细的说明，所属领域的普通技术人员应当理解，依然可以对本发明的具体实施方式进行修改或者对部分技术特征进行等同替换；而不脱离本发明技术方案的精神，其均应涵盖在本发明请求保护的技术方案范围当中。

Claims (9)

1.一种硝基儿茶酚衍生物,它包括如式(Ⅰ)或式(Ⅱ)所示的化合物、其药学上可接受的盐或其溶剂化物，式中，R1选自取代或未取代的酰胺、取代或未取代的C1-6烷基、取代或未取代的C1-6烷氧基、取代或未取代的胺基、取代或未取代的苯基、酯基、氰基、硝基或卤素，所述取代为单取代或多取代，所述取代基为C1-6烷基、卤素、氰基、苯基或硝基；
R2选自H、CN、硝基或卤素。
Figure QLYQS_1
2.根据权利要求1所述的硝基儿茶酚衍生物，其特征在于，R1选自-OCH3、
Figure QLYQS_2
Figure QLYQS_3
其中Y为/>
Figure QLYQS_4
Z为/>
Figure QLYQS_5
Figure QLYQS_6
R2选自H或CN。
3.根据权利要求1所述的硝基儿茶酚衍生物，其特征在于，它选自以下任一种化合物：
Figure QLYQS_7
4.一种根据权利要求1-3任一项所述硝基儿茶酚衍生物的制备方法，其特征在于，包括以下步骤：
Figure QLYQS_8
(1)将化合物1和活性亚甲基发生Knoevenagel缩合反应，得到化合物2；将化合物1用NaClO2发生Pinnick氧化反应得到化合物3；
(2)将化合物2分别与胺、醇进行缩合反应，得到如式(Ⅰ)所示的化合物；将化合物3分别与胺、醇进行缩合反应，得到如式(Ⅱ)所示的化合物。
5.一种药物组合物，其特征在于，它包含权利要求1-3任一项所述硝基儿茶酚衍生物及一种或多种药学上可接受的辅料。
6.根据权利要求5所述的药物组合物，其特征在于，所述药物组合物的剂型为胶囊剂、粉剂、丸剂、片剂、颗粒剂、悬浮剂或注射剂。
7.一种根据权利要求1-3任一项所述硝基儿茶酚衍生物在制备预防和/或治疗帕金森病药物中的应用。
8.一种根据权利要求1-3任一项所述硝基儿茶酚衍生物在制备COMT抑制剂中的应用。
9.一种根据权利要求1-3任一项所述硝基儿茶酚衍生物在制备MAO抑制剂中的应用。
CN202310221983.0A 2023-01-12 2023-03-09 一种硝基儿茶酚衍生物、包含其的药物组合物及其应用 Pending CN116283664A (zh)

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