WO2019196370A1 - 一种五元非芳环并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用 - Google Patents

一种五元非芳环并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用 Download PDF

Info

Publication number
WO2019196370A1
WO2019196370A1 PCT/CN2018/110128 CN2018110128W WO2019196370A1 WO 2019196370 A1 WO2019196370 A1 WO 2019196370A1 CN 2018110128 W CN2018110128 W CN 2018110128W WO 2019196370 A1 WO2019196370 A1 WO 2019196370A1
Authority
WO
WIPO (PCT)
Prior art keywords
bromide
chloride
hiv
membered non
reverse transcriptase
Prior art date
Application number
PCT/CN2018/110128
Other languages
English (en)
French (fr)
Inventor
刘新泳
康东伟
展鹏
武高禅
汪昭
Original Assignee
山东大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 山东大学 filed Critical 山东大学
Priority to US16/765,095 priority Critical patent/US11084830B2/en
Publication of WO2019196370A1 publication Critical patent/WO2019196370A1/zh

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C07D491/048—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being five-membered

Definitions

  • the invention belongs to the technical field of organic compound synthesis and medical application, and particularly relates to a five-membered non-aromatic cyclopyrimidine HIV-1 reverse transcriptase inhibitor, a preparation method and application thereof.
  • HIV-1 human immunodeficiency virus type 1
  • HAART highly active antiretroviral therapy
  • RT inhibitors mainly include Nucleos(t)ide reverse transcriptase inhibitors (N(t)RTIs) and non-nucleoside RT inhibitors (NNRTIs). ).
  • N(t)RTIs Nucleos(t)ide reverse transcriptase inhibitors
  • NRTIs non-nucleoside RT inhibitors
  • NNRTIs are more efficient and less toxic than N(t)RTIs and are an important part of HAART.
  • due to the high variability of HIV-1 virus clinically rapid drug-resistant strains have significantly reduced the clinical efficacy of such drugs.
  • the issue of drug resistance has become an insurmountable gap between anti-AIDS drugs. Therefore, the development of a new generation of highly effective anti-drug resistant NNRTIs is one of the important areas of anti-AIDS drug research.
  • Diarylpyrimidine is a class of HIV-1 NNRTIs with high resistance to drug resistance.
  • the use of diarylpyrimidines as templates for extensive structural modification is of great significance for the discovery of novel anti-HIV drugs with high broad-spectrum anti-drug resistance, good bioavailability and independent intellectual property rights.
  • the present invention provides a five-membered non-aromatic cyclopyrimidine HIV-1 reverse transcriptase inhibitor and a preparation method thereof, and the present invention also provides the above compound as an HIV-1 reverse transcriptase inhibitor Activity screening results and their applications.
  • X is: one of C, N, O, S;
  • Y is: one of C, N, O, S;
  • Z is: one of C, N, O, S;
  • Ar is: phenyl or 4-pyridyl; or SO 2 NH 2 , SO 2 CH 3 , CONH 2 , halogen, NO 2 , CN, NH 2 , CF 3 , NHCH 3 , OH, COOH, CH 2 OH, CO 2 Me, OCH 3 , NHCOCH 3 substituted phenyl; substituents are ortho, meta, para para- or polysubstituted.
  • the five-membered non-aromatic cyclopyrimidine HIV-1 reverse transcriptase inhibitor is one of the following compounds:
  • the "pharmaceutically acceptable salt” as used in the present invention means that within the scope of reliable medical evaluation, the salt of the compound is suitable for contact with tissues of human or lower animals without undue toxicity, irritation and allergy. Reactions, etc., have a fairly reasonable ratio of benefits to risks, usually water or oil soluble or dispersible, and can be effectively used for their intended use.
  • a pharmaceutically acceptable acid addition salt and a pharmaceutically acceptable base addition salt are included herein for the intended use and are compatible with the chemical nature of the compound of formula I. See S. M. Birge et al, J. Pharm. Sci., 1977, 66, pp. 1-19 for a list of suitable salts.
  • a method for preparing a five-membered non-aromatic cyclopyrimidine HIV-1 reverse transcriptase inhibitor comprises the steps of: using a 2,4-dichloro-substituted five-membered non-aromatic ring-pyrimidine 1 as a starting material, first in N,N- The nucleophilic substitution of the substituted phenol or aniline in the dimethylformamide solution to form the intermediate 2; then the nucleophilic substitution reaction of the intermediate 2 with the N-Boc-4-aminopiperidine to form the intermediate 3, and further in the trifluoroacetic acid Removal of Boc protection gives intermediate 4; the last 4 is reacted with various substituted benzyl chloride or benzyl bromide to give the target product 5; the synthetic route is as follows:
  • Reagents and conditions (i) substituted phenol or aniline, N,N-dimethylformamide, potassium carbonate, room temperature; (ii) N-Boc-4-aminopiperidine, N,N-dimethylformamide, Potassium carbonate, 100 ° C; (iii) dichloromethane, trifluoroacetic acid, room temperature; (iv) substituted benzyl chloride or benzyl bromide, N,N-dimethylformamide, potassium carbonate, room temperature.
  • the substituted phenol or aniline is: mesitylene phenol, 2,6-dimethyl-4-cyanophenol, 2,6-dimethyl-4-(E)-cyanovinylphenol, both Trimethylaniline, 2,6-dimethyl-4-cyanoaniline, 2,6-dimethyl-4-(E)-cyanovinylaniline.
  • the substituted benzyl chloride or benzyl bromide is: o-chlorobenzyl chloride, m-chlorobenzyl chloride, p-chlorobenzyl chloride, o-bromo bromide, m-bromo bromide, p-bromo bromide, o-fluorobenzyl bromide, m-chlorobenzyl chloride.
  • the room temperature according to the invention is 20-30 °C.
  • the present invention performs screening of activity of anti-HIV-1 (III B ), double-resistant mutant strain RES056 (K103N/Y181C) at a cellular level on a partial five-membered non-aromatic cyclopyrimidine derivative synthesized according to the above method, and nevirapine (NVP) and etravirine (ETV) were positive controls.
  • the results of the activity are shown in Table 1.
  • Most of the compounds have a single-digit nanomolar inhibitory activity against HIV-1IIIB.
  • the selection index of compounds B2, C1 and C3-4 is more than 100,000, and the safety is extremely high (CC 50 >200 ⁇ M).
  • the five-membered non-aromatic cyclopyrimidine HIV-1 reverse transcriptase inhibitor of the present invention can be used as a non-nucleoside HIV-1 inhibitor. Specifically, it is used as an HIV-1 inhibitor for the preparation of an anti-AIDS drug.
  • An anti-HIV-1 pharmaceutical composition comprising a five-membered non-aromatic cyclopyrimidine HIV-1 reverse transcriptase inhibitor of the invention and one or more pharmaceutically acceptable carriers or excipients.
  • the invention provides a novel five-membered non-aromatic cyclopyrimidine HIV-1 reverse transcriptase inhibitor and a preparation method thereof, and the invention also provides a screening result of the compound for anti-HIV-1 activity and the first time in the field of antiviral application. It has been experimentally proved that the five-membered non-arylcyclopyrimidine derivatives of the present invention can be applied as HIV-1 inhibitors and have high application value. Specifically, it is used as an HIV-1 inhibitor for the preparation of an anti-AIDS drug.
  • the synthesis procedure is the same as 9, except that 4-((2-chloro-5,7-dihydrofuro[3,4-d]pyrimidin-4-yl)oxy)-3,5-dimethyl is selected.
  • the benzonitrile (11) is the starting material.
  • the synthesis procedure is the same as the target compound A1-A6, except that 3,5-dimethyl-4-((2-(piperidin-4-ylamino)-5,7-dihydrofuran[3,4] is used. -d]pyrimidin-4-yl)oxy)benzonitrile (13) is the starting material.
  • the synthesis procedure is the same as the target compound A1-A6, except that 3,5-dimethyl-4-((2-(piperidin-4-ylamino)-5,7-dihydrothieno[3,4] is used. -d]pyrimidin-4-yl)oxy)benzonitrile (21) is the starting material.
  • MTT is a full name of 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide and can be used to detect cell survival and growth.
  • the detection principle is: MTT can be combined with living intracellular succinate dehydrogenase to reduce water-insoluble blue-purple crystal formamidine deposited in cells, while dead cells do not have this function.
  • Dimethyl sulfoxide can dissolve the hyperthyroidism in the cells, and its absorbance (A) value at 540 nm can be indirectly reflected by the microplate reader.
  • the amount of MTT crystal formation is proportional to the number of cells within a certain number of cells.
  • HIV-infected MT-4 cells develop lesions within a certain period of time (5-7 days)
  • HIV-1 (III B ), HIV-2 (ROD) strain, HIV-1 double mutant strain RES056 supplied by the Rega Institute of the University of Leuven, Belgium.
  • NTP nevirapine
  • ETV etravirine
  • Test method The sample was diluted and added to the HIV-infected MT-4 cell suspension. After a period of time, the cell viability was measured by MTT colorimetry, and the absorbance (A) value at 540 nm was recorded in a microplate reader. Calculate EC 50 , CC 50 and SI.
  • MTT colorimetry After adding a sample solution for a period of time, add 20 ⁇ L of MTT solution (5 mg/mL) to each well. After continuing to culture for several hours, discard the staining solution, and add 150 ⁇ L of DMSO to each well, and mix well. The absorbance (A) value at 540 nm was measured by a microplate reader.
  • MT-4 cell culture medium 50 ⁇ L of 1 ⁇ 10 4 MT-4 cell culture medium was added to a 96-well cell culture plate, and 20 ⁇ L of MT-4 cells infected with HIV-1 (III B , single mutant or double mutant) or HIV-2 (ROD) were added. Suspension or blank medium (toxicity determination), adding different concentrations of test compound solution or positive control drug, and setting 3 duplicate wells for each concentration.
  • the cells were cultured in a 5% CO 2 atmosphere at 37 ° C for 5 days, 20 mL (5 mg / mL) of MTT solution was added to each well, and the culture was continued for 2 hours, then DMSO was added, and the absorbance of the reaction solution at 540 nm was measured using a microplate reader.
  • a EC 50 concentration of a compound that inhibits 50% of the virus-induced cell mutating effect or a concentration of a compound that protects 50% of the virus-infected cells from cytopathic effects.
  • b CC 50 The concentration at which 50% of cells that are not infected with HIV develop lesions.
  • c SI selection factor, ratio of CC 50 /EC 50 .

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Virology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Oncology (AREA)
  • Molecular Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicinal Chemistry (AREA)
  • Communicable Diseases (AREA)
  • AIDS & HIV (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

本发明涉及一种五元非芳环并嘧啶类HIV-1逆转录酶抑制剂及其制备方法和应用。所述化合物具有式I所示的结构。本发明还涉及含有式I结构化合物的药物组合物。本发明还提供上述化合物以及含有一个或多个此类化合物的组合物在制备治疗和预防人免疫缺陷病毒(HIV)药物中的应用。

Description

一种五元非芳环并嘧啶类HIV-1逆转录酶抑制剂及其制备方法和应用 技术领域
本发明属于有机化合物合成与医药应用技术领域，具体涉及一种五元非芳环并嘧啶类HIV-1逆转录酶抑制剂及其制备方法和应用。
背景技术
艾滋病(AIDS)是由人体免疫缺陷病毒1型(HIV-1)感染并导致人体防御机能缺陷(尤其是细胞介导的免疫机能缺陷)，而易于发生机会性感染和肿瘤的临床综合征，属于“重大新药创制”科技重大专项列出的10类严重危害我国人民健康的重大疾病之一。目前我国HIV/AIDS流行已经进入快速增长期，感染人数已超过70万。尽管高效抗逆转录疗法(HAART)的普遍实施，使AIDS发病率和死亡率大大降低，但长期服药带来的毒副作用和交叉耐药等问题，使其临床应用受到很大限制。因此，研发新型的抗艾滋病药物依旧是当前一项重大的科研任务。
在HIV-1的生命周期中，逆转录酶(reverse transcriptase，RT)负责将携带病毒遗传信息的单链RNA逆转录成双链DNA，是抗艾滋病药物设计的关键靶标。RT抑制剂主要包括为核苷(酸)类逆转录酶抑制剂(Nucleos(t)ide reverse transcriptase inhibitors,N(t)RTIs)和非核苷类逆转录酶抑制剂(Non-nucleoside RT inhibitors,NNRTIs)。其中，NNRTIs相比于N(t)RTIs具有高效、低毒等优点，是HAART重要组成部分。但由于HIV-1病毒的高变异性，临床上快速出现的耐药株显著地降低了该类药物的临床疗效。耐药性问题成为了抗艾滋病药物难以逾越的鸿沟，因此，新一代高效抗耐药性NNRTIs的研发是目前抗艾滋病药物研究的重要领域之一。
Figure PCTCN2018110128-appb-000001
二芳基嘧啶(diarylpyrimidine,DAPY)类是一类具有高效抗耐药性的HIV-1 NNRTIs，最新一代HIV-1上市药物依曲韦林(Etravirine)和利匹韦林(Rilpivirne)均属于此类化合物。 以二芳基嘧啶类化合物为模板，进行广泛的结构修饰，对发现具有高效广谱抗耐药性、生物利用度好且具有自主知识产权的新型抗HIV药物具有重要意义。
发明内容
针对现有技术的不足，本发明提供了一种五元非芳环并嘧啶类HIV-1逆转录酶抑制剂及其制备方法，本发明还提供上述化合物作为HIV-1逆转录酶抑制剂的活性筛选结果及其应用。
本发明的技术方案如下：
1.五元非芳环并嘧啶类HIV-1逆转录酶抑制剂
一种五元非芳环并嘧啶类HIV-1逆转录酶抑制剂，或其药学上可接受的盐，具有通式I所示的结构：
Figure PCTCN2018110128-appb-000002
其中，
R为：CH 3，CN或者CH＝CHCN；
X为：C，N，O，S中的一种；
Y为：C，N，O，S中的一种；
Z为：C，N，O，S中的一种；
并且X、Y、Z至少有两个同时为C原子；
Ar为：苯基或4-吡啶基；或SO 2NH 2，SO 2CH 3，CONH 2，卤素，NO 2，CN，NH 2，CF 3，NHCH 3，OH，COOH，CH 2OH，CO 2Me，OCH 3，NHCOCH 3取代的苯基；取代基为邻、间、对位单取代或多取代。
根据本发明优选的，五元非芳环并嘧啶类HIV-1逆转录酶抑制剂是下列化合物之一：
Figure PCTCN2018110128-appb-000003
Figure PCTCN2018110128-appb-000004
Figure PCTCN2018110128-appb-000005
本发明中所述的“药学上可接受的盐”是指在可靠的医药评价范围内，化合物的盐类适 于与人或较低等动物的组织相接触而无不适当的毒性、刺激及过敏反应等，具有相当合理的收益与风险比例，通常是水或油可溶的或可分散的，并可有效地用于其预期的用途。包括药学上可接受的酸加成盐和药学上可接受的碱加成盐，在这里是可做预期的用途并与式I化合物的化学性质相容的。适宜的盐的列表参见S.M.Birge等，J.Pharm.Sci.,1977,66,1-19页。
2.五元非芳环并嘧啶类HIV-1逆转录酶抑制剂的制备方法
五元非芳环并嘧啶类HIV-1逆转录酶抑制剂的制备方法，步骤包括：以2,4-二氯取代的五元非芳环并嘧啶1为初始原料，首先在N,N-二甲基甲酰胺溶液中与取代苯酚或苯胺经亲核取代生成中间体2；然后中间体2与N-Boc-4-氨基哌啶发生亲核取代反应生成中间体3，进而在三氟乙酸中脱去Boc保护得到中间体4；最后4经与各种取代氯苄或溴苄的反应生成目标产物5；合成路线如下：
Figure PCTCN2018110128-appb-000006
试剂及条件:(i)取代苯酚或苯胺，N,N-二甲基甲酰胺，碳酸钾，室温；(ii)N-Boc-4-氨基哌啶，N,N-二甲基甲酰胺，碳酸钾，100℃；(iii)二氯甲烷，三氟乙酸，室温；(iv)取代氯苄或溴苄，N,N-二甲基甲酰胺，碳酸钾，室温。
X、Y、Z、R、Ar同上述通式I所示。
所述的取代苯酚或苯胺为：均三甲基苯酚，2,6-二甲基-4-氰基苯酚，2,6-二甲基-4-(E)-氰基乙烯基苯酚，均三甲基苯胺，2,6-二甲基-4-氰基苯胺，2,6-二甲基-4-(E)-氰基乙烯基苯胺。
所述的取代氯苄或溴苄为：邻氯氯苄、间氯氯苄、对氯氯苄、邻溴溴苄、间溴溴苄、对溴溴苄、邻氟氯苄、间氟氯苄、对氟氯苄、2,4-二氟溴苄、3,4-二氟溴苄、邻氰基氯苄、间氰基氯苄、对氰基氯苄、邻硝基氯苄、间硝基氯苄、对硝基氯苄、邻甲氧基氯苄、间甲氧基氯苄、对甲氧基氯苄、对甲磺酰基溴苄、间甲磺酰基溴苄、邻甲磺酰基溴苄、对磺酰胺基溴苄、间磺酰胺基溴苄、邻磺酰胺基溴苄、对甲酰胺基溴苄、间甲酰胺基溴苄、邻甲酰胺基溴苄、4-(溴甲基)苯甲酸甲酯、3-(溴甲基)苯甲酸甲酯、2-(溴甲基)苯甲酸甲酯。
本发明所述的室温为20-30℃。
3.五元非芳环并嘧啶类HIV-1逆转录酶抑制剂的抗HIV-1活性及应用
本发明对按照上述方法合成的部分五元非芳环并嘧啶类衍生物进行了细胞水平的抗 HIV-1(III B)，双耐药突变株RES056(K103N/Y181C)的活性筛选，以奈韦拉平(NVP)和依曲韦林(ETV)为阳性对照。
活性结果如表1所示，多数化合物对HIV-1IIIB具有个位数纳摩尔的抑制活性。A1、A4、B1-4、C1、C3-4和D2的EC 50值在1.1-4.3nM之间，优于ETV(EC 50＝5.1nM)，且选择指数多在10000以上。其中，化合物B2、C1和C3-4的选择指数均大于10万，安全性极高(CC 50>200μM)。活性最高的化合物为C1(EC 50＝1.1nM)，是ETV的4.6倍，且选择指数极高(CC 50>234μM，SI>208333)。对于HIV-1双突变株RES056，C1(EC 50＝25.8nM，SI>9091)同样表现出了最好的抑制活性，是ETV(EC 50＝45.4nM)的1.7倍。此外，A1-2和B1-4对RES056的EC 50值在28.8-43.6nM之间，表现出优于或者与ETV相近的活性。这些结果表明五元非芳环并嘧啶类HIV-1逆转录酶抑制剂具有进一步研究与开发的价值，可作为抗HIV-1的先导化合物加以利用。
本发明的五元非芳环并嘧啶类HIV-1逆转录酶抑制剂可作为非核苷类HIV-1抑制剂应用。具体地说，作为HIV-1抑制剂用于制备抗艾滋病药物。
一种抗HIV-1药物组合物，包括本发明的五元非芳环并嘧啶类HIV-1逆转录酶抑制剂和一种或多种药学上可接受载体或赋形剂。
本发明提供了结构全新的五元非芳环并嘧啶类HIV-1逆转录酶抑制剂及其制备方法，本发明还提供了化合物抗HIV-1活性筛选结果及其在抗病毒领域中的首次应用。经过试验证明，本发明的五元非芳环并嘧啶类衍生物可作为HIV-1抑制剂应用并具有很高的应用价值。具体地说，作为HIV-1抑制剂用于制备抗艾滋病药物。
具体实施方式
通过下述实施例有助于理解本发明，但是不能限制本发明的内容。
实施例中所涉及的合成路线如下：
Figure PCTCN2018110128-appb-000007
Figure PCTCN2018110128-appb-000008
实施例1：4-((2-氯-6,7-二氢噻吩并[3,2-d]嘧啶-4-基)氧基)-3,5-二甲基苄腈(7)的制备
Figure PCTCN2018110128-appb-000009
称取4-羟基-3,5-二甲基苯腈(1.5g,10mmol)和碳酸钾(1.7g,12mmol)于30mL的DMF中，室温搅拌15分钟，然后加入2,4-二氯-6,7-二氢噻吩并[3,2-d]嘧啶6(2.1g,10mmol)继续室温搅拌2h(TLC检测反应完毕)。待有大量白色固体生成，慢慢向其中加入100mL冰水，过滤，真空干燥箱干燥，乙醇中重结晶得中间体7。白色固体，收率94％，熔点250-253℃。 1H NMR(400MHz,DMSO-d 6)δ9.60(s,1H),7.78(s,2H),2.14(s,6H).HRMS:m/z 317.0255[M+1] +.C 14H 9ClN 4OS(316.0186).
实施例2：3,5-二甲基-4-((2-(哌啶-4-基氨基)-6,7-二氢噻吩并[3,2-d]嘧啶-4-基)氧基)苄腈(9)的制备
Figure PCTCN2018110128-appb-000010
依次将7(1.0g,3.17mmol)，N-Boc-4-氨基哌啶(0.83g,3.80mmol)与碳酸钾(0.87g,6.33mmol)加入到5mL的DMF中，然后加热回流10h(TLC检测)。待反应冷却至室温，慢慢将反应液滴加到50mL水中，有大量黄色固体生成。静置30min后过滤，真空干燥得粗品。称取该粗品(1.26g,2.53mmol)溶于4mL二氯甲烷中，加入2.22mL三氟乙酸(30mmol)，室温条件下搅拌3-5h(TLC检测)。然后用饱和的碳酸氢钠溶液调节反应液PH为9，二氯甲烷萃取(3×5mL)，饱和氯化钠溶液洗涤，分取有机层，无水硫酸钠干燥。然后进行快速柱层析分离得中间体9。白色固体，收率64％，熔点130-132℃。 1H NMR(400MHz,DMSO-d 6)δ9.12(s,1H,thiazole-H),8.50(d,J＝5.9Hz,1H,NH),7.67(s,2H,C 3,C 5-Ph-H),3.65–3.62(m,1H),2.78–2.76(m,2H),2.10(s,6H),1.86–1.75(m,2H),1.72–1.55(m,4H).HRMS:m/z 381.1489[M+1] +.C 19H 20N 6OS(380.1419).
实施例3：目标化合物A1-A6的制备
称取化合物9(0.5mmol)于5mL DMF中，然后依次加入碳酸钾(0.14g,1.0mmol)与取代的氯苄或溴苄(0.6mmol)，室温搅拌6-8h(TLC检测)。向反应液中加入饱和氯化钠溶液20mL,乙酸乙酯洗涤(3×10mL)，分取有机层，无水硫酸钠干燥。然后经快速柱层析分离得到目标化合物粗品，进而在乙酸乙酯-石油醚体系中重结晶得到目标化合物A1-A6。
4-((4-((4-(4-氰基-2,6-二甲基苯氧基)-6,7-二氢噻吩并[3,2-d]嘧啶-2-基)氨基)哌啶-1-基)甲基)苯磺酰胺(A1)
Figure PCTCN2018110128-appb-000011
白色固体，收率58％，熔点201-203℃。 1H NMR(400MHz,DMSO-d 6)δ7.77(d,J＝8.0Hz,2H,C 3,C 5-Ph’-H),7.67(s,2H,C 3,C 5-Ph”-H),7.45(d,J＝8.0Hz,2H,C 2,C 6-Ph’-H),7.31(s,2H,SO 2NH 2),7.09(s,1H,NH),3.45(s,2H,N-CH 2),3.37–3.35(m,3H),3.14(t,J＝8.2Hz,2H,S-CH 2),2.71–2.60(m,2H),2.06(s,6H),1.80–1.20(m,6H). 13C NMR(100MHz,DMSO-d 6)δ162.0,160.6,143.3,143.1,133.1,129.4,126.0,119.1,108.6,61.9,52.6,31.5,29.3,16.2.HRMS:m/z 551.1890[M+1] +.C 27H 30N 6O 3S 2(550.1821).
4-((4-((4-(4-氰基-2,6-二甲基苯氧基)-6,7-二氢噻吩并[3,2-d]嘧啶-2-基)氨基)哌啶-1-基)甲基)苯甲酰胺(A2)
Figure PCTCN2018110128-appb-000012
白色固体，收率67％，熔点245-248℃。 1H NMR(400MHz,DMSO-d 6)δ7.92(s,1H),7.82(d,J＝8.0Hz,2H,C 3,C 5-Ph’-H),7.67(s,2H,C 3,C 5-Ph”-H),7.34–7.32(m,3H),6.95(s,1H,NH),3.43(s,2H,N-CH 2),3.39–3.30(m,3H),3.14(t,J＝8.1Hz,2H,S-CH 2),2.67(s,2H),2.06(s,6H),1.78–1.19(m,6H). 13C NMR(101MHz,DMSO-d 6)δ168.2,160.6,142.5,133.4,133.1,132.6,128.8,127.8,108.6,62.2,52.6,36.9,31.5,29.3,16.2.HRMS:m/z 515.2219[M+1] +.C 28H 30N 6O 2S(514.2151).
3,5-二甲基-4-((2-((1-(4-(甲基磺酰基)苄基)哌啶-4-基)氨基)-6,7-二氢噻吩并[3,2-d]嘧啶-4-基)氧基)苯甲腈(A3)
Figure PCTCN2018110128-appb-000013
白色固体，收率70％，熔点142-144℃。 1H NMR(400MHz,DMSO-d 6)δ7.87(d,J＝7.9Hz,2H,C 3,C 5-Ph’-H),7.67(s,2H,C 3,C 5-Ph”-H),7.54(d,J＝8.0Hz,2H,C 2,C 6-Ph’-H),7.10(s,1H,NH),3.50(s,2H,N-CH 2),3.40–3.29(m,3H),3.20(s,3H,SO 2CH 3),3.15(t,J＝8.0Hz,2H,S-CH 2),2.67(s,2H),2.07(s,6H),1.77–1.21(m,6H). 13C NMR(100MHz,DMSO-d 6)δ160.6, 145.4,139.8,133.1,132.7,129.7,127.4,119.1,108.6,61.8,52.6,44.0,31.6,29.3,16.2.HRMS:m/z 550.1946[M+1] +.C 28H 31N 5O 3S 2(549.1868).
3,5-二甲基-4-((2-((1-(吡啶-4-基甲基)哌啶-4-基)氨基)-6,7-二氢噻吩并[3,2-d]嘧啶-4-基)氧基)苯甲腈(A4)
Figure PCTCN2018110128-appb-000014
白色固体，收率54％，熔点140-142℃。 1H NMR(400MHz,DMSO-d 6)δ8.49(d,J＝5.4Hz,2H,C 3,C 5-Ph’-H),7.66(s,2H,C 3,C 5-Ph”-H),7.28(d,J＝5.1Hz,2H,C 2,C 6-Ph’),7.07(s,1H,NH),3.43(s,2H,N-CH 2),3.39–3.25(m,3H),3.15(t,J＝8.1Hz,2H,S-CH 2),2.67(s,2H),2.07(s,6H),1.79–1.23(m,6H). 13C NMR(100MHz,DMSO-d 6)δ162.0,160.6,153.9,149.9,148.1,133.1,132.6,124.1,108.6,61.2,52.6,36.9,31.5,29.3,16.2.HRMS:m/z 473.2118[M+1] +.C 26H 28N 6OS(472.2045).
3,5-二甲基-4-((2-((1-(4-硝基苄基)哌啶-4-基)氨基)-6,7-二氢噻吩并[3,2-d]嘧啶-4-基)氧基)苯甲腈(A5)
Figure PCTCN2018110128-appb-000015
白色固体，收率62％，熔点158-160℃。 1H NMR(400MHz,DMSO-d 6)δ8.19(d,J＝8.3Hz,2H,C 3,C 5-Ph’-H),7.66(s,2H,C 3,C 5-Ph”-H),7.55(d,J＝8.3Hz,2H,C 2,C 6-Ph’-H),7.07(s,1H,NH),3.54(s,2H,N-CH 2),3.39–3.25(m,3H),3.15(t,J＝8.0Hz,2H,S-CH 2),2.68(s,2H),2.07(s,6H),1.82–1.20(m,6H). 13C NMR(100MHz,DMSO-d 6)δ162.0,160.6,147.5,146.9,133.1,132.7,130.0,123.8,119.0,108.6,61.6,52.6,39.4,36.9,31.6,29.3,16.2.HRMS:m/z 517.2016[M+1] +.C 27H 28N 6O 3S(516.1944).
3-((4-((4-(4-氰基-2,6-二甲基苯氧基)-6,7-二氢噻吩并[3,2-d]嘧啶-2-基)氨基)哌啶-1-基)甲基)苯甲酰胺(A6)
Figure PCTCN2018110128-appb-000016
白色固体，收率47％，熔点215-217℃。 1H NMR(400MHz,DMSO-d 6)δ7.95(s,1H,C 2-Ph’-H),7.75(dd,J＝11.0,3.9Hz,2H,C 5,C 6-Ph’-H),7.66(s,2H,C 3,C 5-Ph”-H),7.46–7.25(m,3H),7.06(s,1H,NH),3.43(s,2H,N-CH 2),3.39–3.25(m,3H),3.15(t,J＝8.1Hz,2H,S-CH 2),2.68(s,2H),2.07(s,6H),1.81–1.24(m,6H). 13C NMR(100MHz,DMSO-d 6)δ168.4,162.0,160.6,139.2,134.6,133.1,132.7,132.0,128.4,126.4,119.0,108.6,62.4,52.6,36.9,31.6,29.3,16.2.HRMS:m/z 515.2229[M+1] +.C 28H 30N 6O 2S(514.2151).
实施例4：4-((2-氯-5,7-二氢呋喃并[3,4-d]嘧啶-4-基)氧基)-3,5-二甲基苄腈(11)的制备
Figure PCTCN2018110128-appb-000017
合成步骤同7，所不同的是选用2,4-二氯-5,7-二氢呋喃并[3,4-d]嘧啶(10)为起始原料。白色固体，收率88％，熔点180-183℃。 1H NMR(400MHz,DMSO-d 6)δ7.69(s,2H,C 3,C 5-Ph-H),4.97(s,2H,C 5-dihydrofuropyrimidine),4.80(s,2H,C 7-dihydrofuropyrimidine),2.10(s,6H).HRMS:m/z 302.0687[M+1] +.C 15H 12ClN 3O 2(301.0618).
实施例5：3,5-二甲基-4-((2-(哌啶-4-基氨基)-5,7-二氢呋喃并[3,4-d]嘧啶-4-基)氧基)苄腈(13)的制备
Figure PCTCN2018110128-appb-000018
合成步骤同9，所不同的是选用4-((2-氯-5,7-二氢呋喃并[3,4-d]嘧啶-4-基)氧基)-3,5-二甲基苄腈(11)为起始原料。白色固体，收率61％，熔点122-124℃。 1H NMR(400MHz,DMSO-d 6)δ7.67(s,2H,C 3,C 5-Ph-H),7.08(s,1H,NH),4.96(s,2H,C 5-dihydrofuropyrimidine),4.77(s,2H,C 7-dihydrofuropyrimidine),3.63–3.61(m,1H),2.67(s,2H),2.09(s,6H),1.97–1.32(m,6H).HRMS:m/z 366.1843[M+1] +.C 20H 23N 5O 2(365.1852).
实施例6：目标化合物B1-B6的制备
合成步骤同目标化合物A1-A6，所不同的是选用3,5-二甲基-4-((2-(哌啶-4-基氨基)-5,7-二氢呋喃并[3,4-d]嘧啶-4-基)氧基)苄腈(13)为起始原料。
4-((4-((4-(4-氰基-2,6-二甲基苯氧基)-5,7-二氢呋喃并[3,4-d]嘧啶-2-基)氨基)哌啶-1-基)甲基)苯磺酰胺(B1)
Figure PCTCN2018110128-appb-000019
白色固体，收率66％，熔点191-193℃。 1H NMR(400MHz,DMSO-d 6)δ7.77(d,J＝8.0Hz,2H,C 3,C 5-Ph’-H),7.67(s,2H,C 3,C 5-Ph”-H),7.45(d,J＝8.0Hz,2H,C 2,C 6-Ph’-H),7.31(s,2H,SO 2NH 2),7.08(s,1H,NH),4.96(s,2H,C 5-dihydrofuropyrimidine),4.77(s,2H,C 7-dihydrofuropyrimidine),3.63(s,1H),3.47(s,2H,N-CH 2),2.69(s,2H),2.09(s,6H),1.94–1.18(m,6H). 13C NMR(101MHz,DMSO-d 6)δ162.9,162.0,143.3,143.1,133.1,129.4,126.9,126.2,126.0,119.0,61.9,52.6,31.5,16.2.HRMS:m/z 535.2126[M+1] +.C 27H 30N 6O 4S(534.2049).
4-((4-((4-(4-氰基-2,6-二甲基苯氧基)-5,7-二氢呋喃并[3,4-d]嘧啶-2-基)氨基)哌啶-1-基)甲基)苯甲酰胺(B2)
Figure PCTCN2018110128-appb-000020
白色固体，收率57％，熔点238-240℃。 1H NMR(400MHz,DMSO-d 6)δ7.85(s,2H, CONH 2),7.75(d,J＝7.9Hz,2H,C 3,C 5-Ph’-H),7.60(s,2H,C 3,C 5-Ph”-H),7.26(d,J＝8.1Hz,2H,C 2,C 6-Ph’-H),7.00(s,1H,NH),4.89(s,2H,C 5-dihydrofuropyrimidine),4.70(s,2H,C 7-dihydrofuropyrimidine),3.60(s,1H),3.42(s,2H,N-CH 2),2.74–2.53(m,2H),2.02(s,6H),1.93–1.11(m,6H). 13C NMR(100MHz,DMSO-d 6)δ168.2,162.9,162.5,142.5,133.4,133.1,132.5,128.8,127.8,119.0,108.7,62.2,52.6,31.6,16.2.HRMS:m/z 499.2457[M+1] +.C 28H 30N 6O 3(498.2397).
3,5-二甲基-4-((2-((1-(4-(甲基磺酰基)苄基)哌啶-4-基)氨基)-5,7-二氢呋喃并[3,4-d]嘧啶-4-基)氧基)苄腈(B3)
Figure PCTCN2018110128-appb-000021
白色固体，收率72％，熔点209-211℃。 1H NMR(400MHz,DMSO-d 6)δ7.87(d,J＝8.0Hz,2H,C 3,C 5-Ph’-H),7.67(s,2H,C 3,C 5-Ph”-H),7.54(d,J＝8.0Hz,2H,C 2,C 6-Ph’-H),7.08(s,1H,NH),4.96(s,2H,C 5-dihydrofuropyrimidine),4.77(s,2H,C 7-dihydrofuropyrimidine),3.68(s,1H),3.51(s,2H,N-CH 2),3.20(s,3H,SO 2CH 3),2.76–2.62(m,2H),2.09(s,6H),1.90–1.15(m,6H). 13C NMR(101MHz,DMSO-d 6)δ162.9,162.5,145.4,139.8,133.1,129.7,127.4,108.7,69.3,61.8,52.6,44.0,31.5,16.2.HRMS:m/z 534.2173[M+1] +.C 28H 31N 5O 4S(533.2097).
3,5-二甲基-4-((2-((1-(吡啶-4-基甲基)哌啶-4-基)氨基)-5,7-二氢呋喃并[3,4-d]嘧啶-4-基)氧基)苄腈(B4)
Figure PCTCN2018110128-appb-000022
白色固体，收率73％，熔点155-157℃。 1H NMR(400MHz,DMSO-d 6)δ8.50(d,J＝5.9Hz,2H,C 3,C 5-Ph’-H),7.67(s,2H,C 3,C 5-Ph”-H),7.28(d,J＝5.1Hz,2H,C 2,C 6-Ph’-H),7.09(s,1H,NH),4.96(s,2H,C 5-dihydrofuropyrimidine),4.77(s,2H,C 7-dihydrofuropyrimidine),3.68(s,1H),3.45(s,2H,N-CH 2),2.77–2.59(m,2H),2.09(s,6H),1.91–1.22(m,6H). 13C NMR(100MHz, DMSO-d 6)δ162.9,149.9,148.1,133.1,124.1,108.7,69.4,61.2,52.6,31.4,16.2.HRMS:m/z457.2345[M+1] +.C 26H 28N 6O 2(456.2274).
3,5-二甲基-4-((2-((1-(4-硝基苄基)哌啶-4-基)氨基)-5,7-二氢呋喃并[3,4-d]嘧啶-4-基)氧基)苄腈(B5)
Figure PCTCN2018110128-appb-000023
白色固体，收率61％，熔点199-201℃。 1H NMR(400MHz,DMSO-d 6)δ8.12(d,J＝8.6Hz,2H,C 3,C 5-Ph’-H),7.60(s,2H,C 3,C 5-Ph”-H),7.49(d,J＝8.4Hz,2H,C 2,C 6-Ph’-H),7.02(s,1H,NH),4.89(s,2H,C 5-dihydrofuropyrimidine),4.70(s,2H,C 7-dihydrofuropyrimidine),3.61(s,1H),3.47(s,2H,N-CH 2),2.63(s,2H),2.02(s,6H),1.80–1.14(m,6H). 13C NMR(100MHz,DMSO-d 6)δ162.9,146.9,133.1,130.0,123.8,61.6,52.6,39.6,39.4,31.5,16.2.HRMS:m/z501.2247[M+1] +.C 27H 28N 6O 4(500.2172).
3-((4-((4-(4-氰基-2,6-二甲基苯氧基)-5,7-二氢呋喃并[3,4-d]嘧啶-2-基)氨基)哌啶-1-基)甲基)苯甲酰胺(B6)
Figure PCTCN2018110128-appb-000024
白色固体，收率43％，熔点225-227℃。 1H NMR(400MHz,DMSO-d 6)δ7.88(s,2H,CONH 2),7.70(s,1H,C 2-Ph’-H),7.69–7.66(m,1H),7.60(s,2H,C 3,C 5-Ph”-H),7.35–7.30(m,2H,C 2,C 6-Ph’-H),7.01(s,1H,NH),4.89(s,2H,C 5-dihydrofuropyrimidine),4.70(s,2H,C 7-dihydrofuropyrimidine),3.60(s,1H),3.36(s,2H,N-CH 2),2.63(s,2H),2.02(s,6H),1.92–1.16(m,6H). 13C NMR(101MHz,DMSO-d 6)δ168.4,163.0,162.5,139.2,134.6,133.1,132.0,128.4,126.4,119.0,108.7,69.4,62.4,55.3,52.6,39.6,31.6,16.2.HRMS:m/z 499.2456[M+1] +.C 28H 30N 6O 3(498.2379).
实施例7：4-((2-氯-6,7-二氢-5H-环戊二烯并[d]嘧啶-4-基)氧基)-3,5-二甲基苄腈(15)的制备
Figure PCTCN2018110128-appb-000025
合成步骤同7，所不同的是选用2,4-二氯-6,7-二氢-5H-环戊二烯并[d]嘧啶(14)为起始原料。白色固体，收率97％，熔点254-256℃。 1H NMR(400MHz,DMSO-d 6)δ7.73(s,2H,C 3,C 5-Ph-H),3.04(t,J＝7.8Hz,2H),2.92(t,J＝7.7Hz,2H),2.13(p,J＝7.7Hz,2H),2.09(s,6H).HRMS:m/z 300.0895[M+1] +.C 16H 14ClN 3O(299.0825).
实施例8：3,5-二甲基-4-((2-(哌啶-4-基氨基)-6,7-二氢-5H-环戊二烯并[d]嘧啶-4-基)氧基)苄腈(17)的制备
Figure PCTCN2018110128-appb-000026
合成步骤同9，所不同的是选用4-((2-氯-6,7-二氢-5H-环戊二烯并[d]嘧啶-4-基)氧基)-3,5-二甲基苄腈(15)为起始原料。白色固体，收率67％，熔点135-137℃。 1H NMR(400MHz,DMSO-d 6)δ7.64(s,2H,C 3,C 5-Ph-H),6.90(s,1H,NH),3.65(s,1H),2.76–2.72(m,4H),2.69–2.59(m,2H),2.07(s,6H),2.02–1.97(m,2H),1.81–1.21(m,6H).HRMS:m/z 364.2132[M+1] +.C 21H 25N 5O(363.2059).
实施例9：目标化合物C1-C6的制备
合成步骤同目标化合物A1-A6，3,5-二甲基-4-((2-(哌啶-4-基氨基)-6,7-二氢-5H-环戊二烯并[d]嘧啶-4-基)氧基)苄腈(17)为起始原料。
4-((4-((4-(4-氰基-2,6-二甲基苯氧基)-6,7-二氢-5H-环戊二烯并[d]嘧啶-2-基)氨基)哌啶-1-基)甲基)苯磺酰胺(C1)
Figure PCTCN2018110128-appb-000027
白色固体，收率80％，熔点207-209℃。 1H NMR(400MHz,DMSO-d 6)δ7.77(d,J＝8.3Hz,2H,C 3,C 5-Ph’-H),7.64(s,2H,C 3,C 5-Ph”-H),7.45(d,J＝8.1Hz,2H,C 2,C 6-Ph’-H),7.30(s,2H,SO 2NH 2),6.90(s,1H,NH),3.65(s,1H),3.45(s,2H),2.76(dt,J＝22.3,7.7Hz,4H),2.69–2.59(m,2H),2.07(s,6H),2.02(d,J＝7.4Hz,2H),1.81–1.21(m,6H). 13C NMR(100MHz,DMSO-d 6)δ162.0,143.3,143.1,133.1,132.6,129.4,126.0,119.1,108.3,62.0,52.6,39.6,31.6,25.9,22.0,16.3.HRMS:m/z 533.2330[M+1] +.C 28H 32N 6O 3S(532.2257).
4-((4-((4-(4-氰基-2,6-二甲基苯氧基)-6,7-二氢-5H-环戊二烯并[d]嘧啶-2-基)氨基)哌啶-1-基)甲基)苯甲酰胺(C2)
Figure PCTCN2018110128-appb-000028
白色固体，收率68％，熔点225-227℃。 1H NMR(400MHz,DMSO-d 6)δ7.90(s,2H,CONH 2),7.81(d,J＝7.9Hz,2H,C 3,C 5-Ph’-H),7.64(s,2H,C 3,C 5-Ph”-H),7.32(d,J＝8.0Hz,2H,C 2,C 6-Ph’-H),6.89(s,1H,NH),3.65(s,1H),3.48–3.36(m,2H),2.76(dt,J＝22.1,7.7Hz,4H),2.70–2.58(m,2H),2.07(s,6H),2.02(d,J＝7.4Hz,2H),1.70–1.23(m,6H). 13C NMR(100MHz,DMSO-d 6)δ168.2,162.0,143.1,142.5,133.4,133.1,129.5,128.8,127.8,119.5,108.9,62.2,52.7,39.0,31.2,25.9,22.0,16.3.HRMS:m/z 497.2657[M+1] +.C 29H 32N 6O 2(496.2587).
3,5-二甲基-4-((2-((1-(4-(甲基磺酰基)苄基)哌啶-4-基)氨基)-6,7-二氢-5H-环戊二烯并[d]嘧啶-4-基)氧基)苄腈(C3)
Figure PCTCN2018110128-appb-000029
白色固体，收率69％，熔点170-172℃。 1H NMR(400MHz,DMSO-d 6)δ7.89–7.85(m,2H),7.65(s,2H,C 3,C 5-Ph”-H),7.54(d,J＝8.1Hz,2H),6.80(s,1H,NH),3.66(s,1H),3.50(s,2H,N-CH 2),2.89(s,3H),2.78(dd,J＝14.7,7.3Hz,4H),2.71–2.61(m,2H),2.07(s,6H),2.03(d,J＝7.5Hz,2H),1.81–1.23(m,6H). 13C NMR(100MHz,DMSO-d 6)δ162.7,162.0,145.4,139.8,133.1,132.6,129.7,127.4,127.3,108.3,62.6,61.9,52.6,44.1,44.0,36.2,31.6,31.2,25.9,22.0,16.3.HRMS:m/z 532.2377[M+1] +.C 29H 33N 5O 3S(531.2304).
3,5-二甲基-4-((2-((1-(吡啶-4-基甲基)哌啶-4-基)氨基)-6,7-二氢-5H-环戊二烯并[d]嘧啶-4-基)氧基)苄腈(C4)
Figure PCTCN2018110128-appb-000030
白色固体，收率72％，熔点173-175℃。 1H NMR(400MHz,DMSO-d 6)δ8.57(d,J＝7.9Hz,2H,C 3,C 5-Ph’-H),7.64(s,2H,C 3,C 5-Ph”-H),7.28(d,J＝8.0Hz,2H,C 2,C 6-Ph’-H),6.83(s,1H,NH),3.60(s,1H),3.48–3.42(m,2H),2.76(dt,J＝21.6,7.2Hz,4H),2.70–2.62(m,2H),2.07(s,6H),2.02(d,J＝7.4Hz,2H),1.85–1.21(m,6H). 13C NMR(100MHz,DMSO-d 6)δ168.2,162.4,142.9,142.1,133.7,133.2,129.5,127.8,1120.3,108.7,62.2,52.6,39.0,31.6,25.9,22.0,16.3.HRMS:m/z 455.2557[M+1] +.C 27H 30N 6O(454.2481).
3,5-二甲基-4-((2-((1-(4-硝基苄基)哌啶-4-基)氨基)-6,7-二氢-5H-环戊二烯并[d]嘧啶-4-基)氧基)苄腈(C5)
Figure PCTCN2018110128-appb-000031
白色固体，收率71％，熔点200-212℃。 1H NMR(400MHz,DMSO-d 6)δ8.17(d,J＝8.1Hz,2H,C 3,C 5-Ph’-H),7.65(s,2H,C 3,C 5-Ph”-H),7.54(d,J＝8.1Hz,2H),6.80(s,1H,NH),3.72(s,1H),3.50(s,2H,N-CH 2),2.78(dd,J＝14.4,7.2Hz,4H),2.71–2.64(m,2H),2.07(s,6H),2.03(d,J＝7.5Hz,2H),1.81–1.23(m,6H). 13C NMR(100MHz,DMSO-d 6)δ159.7,147.5,146.9,145.4,139.8,133.1,132.6,130.2,127.4,123.8,108.3,62.1,61.7,52.6,44.1,36.2,31.6,31.2,25.9,21.8,16.2.HRMS:m/z 499.2456[M+1] +.C 28H 30N 6O 3(498.2379).
3-((4-((4-(4-氰基-2,6-二甲基苯氧基)-6,7-二氢-5H-环戊二烯并[d]嘧啶-2-基)氨基)哌啶-1-基)甲基)苯甲酰胺(C6)
Figure PCTCN2018110128-appb-000032
白色固体，收率59％，熔点234-236℃。 1H NMR(400MHz,DMSO-d 6)δ7.96(s,2H,CONH 2),7.79–7.75(m,1H),7.65(s,2H,C 3,C 5-Ph”-H),7.39–7.35(m,3H),6.80(s,1H,NH),3.72(s,1H),3.50(s,2H,N-CH 2),2.78(dd,J＝14.3,7.2Hz,4H),2.71–2.63(m,2H),2.07(s,6H),2.03(d,J＝7.5Hz,2H),1.80–1.18(m,6H). 13C NMR(100MHz,DMSO-d 6)δ168.2,162.3,143.2,142.5,133.4,133.1,129.4,128.8,127.6,119.5,108.9,62.2,52.7,39.0,31.3,25.9,22.0,16.3.ESI-MS:m/z 497.2657[M+1] +.C 29H 32N 6O 2(496.2587).
实施例10：4-((2-氯-5,7-二氢噻吩并[3,4-d]嘧啶-4-基)氧基)-3,5-二甲基苄腈(19)的制备
Figure PCTCN2018110128-appb-000033
合成步骤同7，所不同的是选用2,4-二氯-5,7-二氢噻吩并[3,4-d]嘧啶(18)为起始原料。白色固体，收率88％，熔点268-270℃。 1H NMR(400MHz,DMSO-d 6):δ7.72(s,2H,C 3,C 5-Ph-H),4.12(s,2H,S-CH 2),4.07(s,2H,S-CH 2),2.10(s,6H).ESI-MS:m/z 318.2[M+1] +.C 15H 12ClN 3OS(317.04).
实施例11：3,5-二甲基-4-((2-(哌啶-4-基氨基)-5,7-二氢噻吩并[3,4-d]嘧啶-4-基)氧基)苄腈(21)的制备
Figure PCTCN2018110128-appb-000034
合成步骤同9，所不同的是选用4-((2-氯-5,7-二氢[3,4-d]嘧啶-4-基)氧基)-3,5-二甲基苄腈(19)为起始原料。白色固体，收率96％，熔点268-270℃。 1H NMR(400MHz,DMSO-d 6):δ7.72(s,2H,C 3,C 5-Ph-H),6.89(s,1H,NH),4.12(s,2H,S-CH 2),4.08(s,2H,S-CH 2),3.72-3.70(m,1H),2.74-2.72(m,2H),2.10(s,6H),1.97-1.75(m,4H),1.47-1.42(m,2H).ESI-MS:m/z 382.2[M+1] +.C 20H 23N 5OS(381.16).
实施例12：目标化合物D1-D2的制备
合成步骤同目标化合物A1-A6，所不同的是选用3,5-二甲基-4-((2-(哌啶-4-基氨基)-5,7-二氢噻吩并[3,4-d]嘧啶-4-基)氧基)苄腈(21)为起始原料。
4-((4-((4-(4-氰基-2,6-二甲基苯氧基)-5,7-二氢噻吩并[3,4-d]嘧啶-2-基)氨基)哌啶-1-基)甲基)苯磺酰胺(D1)
Figure PCTCN2018110128-appb-000035
白色固体，收率79％，熔点144-146℃。 1H NMR(400MHz,DMSO-d 6):δ7.78(d,J＝8.2Hz,2H,C 3,C 5-Ph’-H),7.72(s,2H,C 3,C 5-Ph”-H),7.46(d,J＝8.1Hz,2H,C 2,C 6-Ph’-H),7.30(s,2H,SO 2NH 2),6.89(s,1H,NH),4.12(s,2H,S-CH 2),4.08(s,2H,S-CH 2),3.72(s,1H),3.42(s,2H,N-CH 2),2.72-2.74(m,2H),2.10(s,6H),1.79-2.04(m,4H),1.41-1.47(m,2H). 13C NMR(100MHz,DMSO-d 6):δ165.7,161.7,160.3,153.7,143.0,135.3,134.2,126.0,123.9,123.7,119.2,109.7,61.2,52.8,48.3,31.7,16.2.HRMS:m/z 551.1890[M+1] +.C 27H 30N 6O 3S 2(550.1821).
3,5-二甲基-4-((2-((1-(4-(甲基磺酰基)苄基)哌啶-4-基)氨基)-5,7-二氢噻吩并[3,4-d]嘧啶-4- 基)氧基)苄腈(D2)
Figure PCTCN2018110128-appb-000036
白色固体，收率71％，熔点181-183℃。 1H NMR(400MHz,DMSO-d 6,ppm):δ7.84(d,2H,J＝8.2Hz,C 3,C 5-Ph’-H),7.71(s,2H,C 3,C 5-Ph”-H),7.52(d,2H,J＝8.2Hz,C 2,C 6-Ph’-H),6.88(s,1H,NH),4.12(s,2H,S-CH 2),4.08(s,2H,S-CH 2),3.75-3.78(m,1H),3.57(s,2H,N-CH 2),3.06(s,3H,SO 2CH 3),2.71-2.75(m,2H),2.10(s,6H),1.96-2.09(m,4H),1.39-1.41(m,2H). 13C NMR(100MHz,DMSO-d6):δ165.1,162.3,159.4,153.9,145.2,139.5,134.3,133.0,126.8,123.3,118.9,109.7,62.3,52.6,48.5,44.7,32.0,16.6.HRMS:m/z 550.1942[M+1] +.C 28H 31N 5O 3S 2(549.1868).
实施例13：目标化合物的体外抗HIV活性测试实验
测试原理：
化合物体外抗HIV活性筛选采用MTT法。MTT全称为溴化-3-(4,5-二甲基-2-噻唑基)-2,5-二苯基四唑氮，可用于检测细胞的存活和生长。检测原理为：MTT可以与活的细胞内琥珀酸脱氢酶结合还原为水不溶性的蓝紫色结晶甲瓒沉积在细胞中，而死细胞并无此功能。二甲基亚砜可以溶解细胞中的甲瓒，用酶标仪检测其在540nm下的吸光度(A)值可以间接的反映活细胞的数量。在一定的细胞数范围内，MTT结晶形成的量与细胞数成正比。
由于HIV感染的MT-4细胞在一定时间内(5-7天)会发生病变，因此向HIV感染的MT-4细胞悬浊液中加入适当浓度的待检测化合物溶液，经过一段时间(5-7天)的培养后，用MTT分析法测定MT-4细胞活力，得到保护50％细胞免于细胞病变的药物浓度EC 50，同时得到目标化合物使50％未感染HIV的细胞发生病变的浓度CC 50，计算出选择系数SI(SI＝CC 50/EC 50)。
测试材料和方法：
(1)HIV-1(III B)、HIV-2(ROD)毒株、HIV-1双突变株RES056：由比利时鲁汶大学医学院Rega研究所提供。
(2)MT-4细胞：由比利时鲁汶大学医学院Rega研究院提供。
(3)MTT：购自美国Sigma公司。
(4)样品处理：样品临用前溶于DMSO配成适当浓度，并用双蒸水作5倍稀释，各5个稀 释度。
(5)阳性对照药：奈韦拉平(NVP)和依曲韦林(ETV)。
(6)测试方法：样品稀释后加入到HIV感染MT-4细胞悬浊液中，经过一段时间后用MTT比色法测定细胞活力，用酶标仪中记录在540nm下的吸光度(A)值,计算出EC 50，CC 50以及SI。
(7)MTT比色法：加入样品溶液培养一段时间后，向每孔加入MTT溶液(5mg/mL)20μL，继续培养若干小时后，弃染色液，并向每孔加入150μL DMSO，充分混合，用酶标仪中测定540nm下的吸光度(A)值。
实验方法：
50μL含1×10 4MT-4细胞培养液加入96孔细胞培养板，加入20μL感染HIV-1(III B、单突变株或双突变株)或HIV-2(ROD)的MT-4细胞混悬液或者空白培养基(毒性测定)，加入不同浓度的待测化合物溶液或者阳性对照药物，每个浓度设3个复孔。细胞在5％CO 2氛围，37℃下培养5天，向每孔加入20mL(5mg/mL)MTT溶液，继续培养2小时，然后加入DMSO，使用酶标仪测定反应溶液在540nm处的吸收度，计算化合物不同浓度下的细胞增值率P％。同时设空白和阳性药物对照组，由此计算化合物保护50％的细胞免于HIV诱导的细胞病变所需浓度(EC 50)，使50％未感染HIV的细胞发生病变的浓度(CC 50)。选择指数的计算：SI＝CC 50/EC 50。
按照上述实验方法对合成的部分五元非芳环并[3,2-d]嘧啶类衍生物进行了细胞水平的抗HIV-1(III B)以及双突变株RES056(K103N/Y181C)的活性筛选，活性结果如表1所示。
表1.部分五元非芳环并嘧啶类类化合物抗HIV活性、毒性及选择指数
Figure PCTCN2018110128-appb-000037
Figure PCTCN2018110128-appb-000038
aEC 50:抑制50％的病毒诱导的致细胞突变效应的化合物浓度或保护50％感染病毒的细胞免于细胞病变的化合物浓度。
bCC 50：使50％未感染HIV的细胞发生病变的浓度。
cSI:选择系数，CC 50/EC 50的比值。

Claims (5)

  1. 一种五元非芳环并嘧啶类HIV-1逆转录酶抑制剂，或其药学上可接受的盐，其特征在于，具有通式I所示的结构：
    Figure PCTCN2018110128-appb-100001
    其中，
    R为：CH 3，CN或者CH＝CHCN；
    X为：C，N，O，S中的一种；
    Y为：C，N，O，S中的一种；
    Z为：C，N，O，S中的一种；
    并且X、Y、Z至少有两个同时为C原子；
    Ar为：苯基或4-吡啶基；或SO 2NH 2，SO 2CH 3，CONH 2，卤素，NO 2，CN，NH 2，CF 3，NHCH 3，OH，COOH，CH 2OH，CO 2Me，OCH 3，NHCOCH 3取代的苯基；取代基为邻、间、对位单取代或多取代。
  2. 如权利要求1所述的五元非芳环并嘧啶类HIV-1逆转录酶抑制剂，其特征在于，是下列化合物之一：
    Figure PCTCN2018110128-appb-100002
    Figure PCTCN2018110128-appb-100003
    Figure PCTCN2018110128-appb-100004
  3. 如权利要求1所述的五元非芳环并嘧啶类HIV-1逆转录酶抑制剂的制备方法，步骤包括：以2,4-二氯取代的五元非芳环并嘧啶1为初始原料，首先在N,N-二甲基甲酰胺溶液中与取代苯酚或苯胺经亲核取代生成中间体2；然后中间体2与N-Boc-4-氨基哌啶发生亲核取代反应生成中间体3，进而在三氟乙酸中脱去Boc保护得到中间体4；最后4经与各种取代氯苄或溴苄的反应生成目标产物5；合成路线如下：
    Figure PCTCN2018110128-appb-100005
    试剂及条件:(i)取代苯酚或苯胺，N,N-二甲基甲酰胺，碳酸钾，室温；(ii)N-Boc-4-氨基哌啶，N,N-二甲基甲酰胺，碳酸钾，100℃；(iii)二氯甲烷，三氟乙酸，室温；(iv)取代氯苄或溴苄，N,N-二甲基甲酰胺，碳酸钾，室温；
    X、Y、Z、R、Ar同权利要求1中通式I所示；
    所述的取代苯酚或苯胺为：均三甲基苯酚，2,6-二甲基-4-氰基苯酚，2,6-二甲基-4-(E)-氰基乙烯基苯酚，均三甲基苯胺，2,6-二甲基-4-氰基苯胺，2,6-二甲基-4-(E)-氰基乙烯基苯胺；
    所述的取代氯苄或溴苄为：邻氯氯苄、间氯氯苄、对氯氯苄、邻溴溴苄、间溴溴苄、对溴溴苄、邻氟氯苄、间氟氯苄、对氟氯苄、2,4-二氟溴苄、3,4-二氟溴苄、邻氰基氯苄、间氰基氯苄、对氰基氯苄、邻硝基氯苄、间硝基氯苄、对硝基氯苄、邻甲氧基氯苄、间甲氧基氯苄、对甲氧基氯苄、对甲磺酰基溴苄、间甲磺酰基溴苄、邻甲磺酰基溴苄、对磺酰胺基溴苄、间磺酰胺基溴苄、邻磺酰胺基溴苄、对甲酰胺基溴苄、间甲酰胺基溴苄、邻甲酰胺基溴苄、4-(溴甲基)苯甲酸甲酯、3-(溴甲基)苯甲酸甲酯、2-(溴甲基)苯甲酸甲酯。
  4. 一种如权利要求1或2所述五元非芳环并嘧啶类HIV-1逆转录酶抑制剂在制备治疗和预防人免疫缺陷病毒(HIV)药物中的应用。
  5. 一种药物组合物，包含权利要求1或2所述五元非芳环并嘧啶类HIV-1逆转录酶抑制剂和一种或多种药学上可接受载体或赋形剂。
PCT/CN2018/110128 2018-04-12 2018-10-12 一种五元非芳环并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用 WO2019196370A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US16/765,095 US11084830B2 (en) 2018-04-12 2018-10-12 Five-membered non aromatic ring-pyrimidine HIV-1 reverse transcriptase inhibitor, preparation method therefor, and uses thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810336405.0 2018-04-12
CN201810336405.0A CN108218890B (zh) 2018-04-12 2018-04-12 一种五元非芳环并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用

Publications (1)

Publication Number Publication Date
WO2019196370A1 true WO2019196370A1 (zh) 2019-10-17

Family

ID=62657945

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2018/110128 WO2019196370A1 (zh) 2018-04-12 2018-10-12 一种五元非芳环并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用

Country Status (3)

Country Link
US (1) US11084830B2 (zh)
CN (1) CN108218890B (zh)
WO (1) WO2019196370A1 (zh)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108218890B (zh) * 2018-04-12 2020-03-27 山东大学 一种五元非芳环并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用
CN110066273A (zh) * 2019-06-05 2019-07-30 山东大学 一种含三氮唑环的单芳基嘧啶类hiv-1逆转录酶抑制剂及其制备方法与应用
CN111217833B (zh) * 2020-02-21 2021-03-16 山东大学 噻吩并[2,3-d]嘧啶类HIV-1非核苷类逆转录酶抑制剂及其制备方法和应用
CN111793074B (zh) * 2020-07-23 2021-10-26 山东大学 K-5a2化合物的晶型及其制备方法和应用
CN114014866A (zh) * 2021-11-09 2022-02-08 山东大学 一种5,7-二氢呋喃并[3,4-d]嘧啶类化合物及其制备方法和应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108218890A (zh) * 2018-04-12 2018-06-29 山东大学 一种五元非芳环并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104530078B (zh) * 2015-01-27 2017-03-22 山东大学 一种噻吩并[3,2‑d]嘧啶衍生物及其制备方法与应用
CN104926829A (zh) * 2015-06-08 2015-09-23 山东大学 一种噻吩并嘧啶类衍生物及其制备方法和应用
CN106117242B (zh) * 2016-06-27 2018-08-03 山东大学 四氢噻喃并嘧啶类衍生物及其制备方法与应用

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108218890A (zh) * 2018-04-12 2018-06-29 山东大学 一种五元非芳环并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用

Also Published As

Publication number Publication date
US11084830B2 (en) 2021-08-10
US20200361954A1 (en) 2020-11-19
CN108218890B (zh) 2020-03-27
CN108218890A (zh) 2018-06-29

Similar Documents

Publication Publication Date Title
WO2019196370A1 (zh) 一种五元非芳环并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用
CA2209018C (en) Sulfonamides and their use
TWI304061B (en) Nitrogen-containing aromatic ring derivatives
WO2016197589A1 (zh) 一种噻吩并嘧啶类衍生物及其制备方法和应用
AU2006229993B2 (en) 8-phenyl-7,8-dihydropyrido(2,3-d)pyrimidin-7-one and their use as pharmaceuticals
US20080300245A1 (en) Pyrazolo[1,5-A]Pyridine-3-Carboxylic Acids as Ephb and Vegfr2 Kinase Inhibitors
US20140221371A1 (en) Heterocyclic compounds and their use as binding partners for 5-ht5 receptors
JP2013516420A (ja) 置換されたピロロ−アミノピリミジン化合物
MX2007011883A (es) Compuestos novedosos.
CN106117242B (zh) 四氢噻喃并嘧啶类衍生物及其制备方法与应用
CN108250122A (zh) 磺酰胺-芳基酰胺类化合物及其治疗乙型肝炎的药物用途
JP6779867B2 (ja) ピリミジン化合物およびその使用方法
CN105175414A (zh) 咪唑[4,5-b]吡啶巯乙酰胺类衍生物及其制备方法与应用
WO2019196369A1 (zh) 一种噻唑并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用
CN108440559B (zh) 二芳基噻吩并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用
US20230067045A1 (en) Thiophene[2,3-d]pyrimidine derivative dk6-1 and its preparation method and application
Mishra et al. Search for new pharmacophores for antimalarial activity. Part III: synthesis and bioevaluation of new 6-thioureido-4-anilinoquinazolines
CN109369623B (zh) 一种取代1,2,3三氮唑类二芳基嘧啶衍生物及其制备方法与应用
US20090118277A1 (en) 3 Unsubstituted N-(aryl- or heteroaryl)-pyrazolo[1,5-a]pyrimidines as Kinase Inhibitors
CN108586482A (zh) 一种含三氮唑环的二芳基嘧啶类hiv-1抑制剂及其制备方法和应用
CN108409734A (zh) 吡啶并嘧啶类hiv-1逆转录酶抑制剂及其制备方法和应用
WO2014134750A1 (zh) 2,6,9-三取代嘌呤衍生物及其制备方法与应用
CN108440500B (zh) 一种喹唑啉类hiv-1抑制剂及其制备方法和应用
RU2246496C1 (ru) Вещество с противовирусной и антибактериальной активностью на основе производных 2,8-дитиоксо-1h-пирано[2,3-d, 6,5-d`] дипиримидина и их 10-аза-аналогов
CN106831814B (zh) 一种噻吩并[3,2-d]嘧啶类HIV-1逆转录酶抑制剂及其制备方法和应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18914862

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18914862

Country of ref document: EP

Kind code of ref document: A1

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 11/05/2021)

122 Ep: pct application non-entry in european phase

Ref document number: 18914862

Country of ref document: EP

Kind code of ref document: A1