呋喃[2,3-d]嘧啶核苷的化学合成 及抗水痘带状疱疹病毒活性的研究进展

摘要：呋喃[2,3-d]嘧啶核苷是一类新型的、特异性治疗水痘带状疱疹病毒的核苷前药。该文综述了近年来呋喃[2,3-d]嘧啶核苷衍生物的化学合成及其抗水痘带状疱疹病毒(varicella-zoster virus , VZV)活性研究进展。     

吡啶[2,3-d)嘧啶类抗菌物质的研究

本文报道了一系列2,6-取代的5,8-二氢-8-乙基-5-氧吡啶[2,3-d]嘧啶类化合物的合成和抗菌活性,并确证2-氯-4-(2’-甲氧羰基二乙氨基)嘧啶-5-羧酸乙酯Ⅳ经过Dieckmann环化所得的产物为2-氯-7,8-二氢-8-乙基-5-羟基吡啶[2,3-d]嘧啶-6-羧酸甲酯Ⅴ,即以烯醇式存在,而不是以Pesson等氏报道的酮式结构Ⅵ存在。     

7-(4-烃氧基苯基)-4-(4-芳基哌嗪-1-基)-5,6,7,8-四氢苯并[4,5]噻吩并[2,3-d]嘧啶类化合物的合成与抗肿瘤活性研究

目的设计合成一系列7-(4-烃氧基苯基)-4-(4-芳基哌嗪-1-基)-5,6,7,8-四氢苯并[4,5]噻吩并[2,3-d]嘧啶类化合物并进行抗肿瘤活性研究。方法以4-(4-羟基苯基)环己酮、氰乙酰胺、单质硫作为起始原料,通过Gewald反应、改进后的Niementowski喹唑啉缩合反应、三氯氧磷氯代反应、亲核取代反应、脱乙基反应及Williamson反应得到相应的目标化合物,采用MTT法测定目标化合物体外抑制人类肺癌细胞株A549增殖的活性,并初步探究其构效关系。结果与结论目标化合物8a、8c、8f和8k在10μmol·L~(-1)浓度下对肿瘤细胞的抑制率达到30%以上,有进一步研究的价值。     

瑞博西尼的微波辅助合成新工艺

目的寻找抗肿瘤新药瑞博西尼的高效合成新工艺。方法首先采用一锅反应制备带有瑞博西尼C-6和N-7两个取代基团的2-环戊氨基-N,N-二甲基乙酰胺;然后,在微波辅助加热条件下,与起始物4-氯-2-甲硫基-嘧啶-5-醛经过第二个一锅反应构建吡咯并嘧啶骨架中间体;该中间体先后经过氧化、强碱条件下的芳香亲核取代反应和最后脱保护得到目标化合物瑞博西尼。结果该工艺虽然包括7步反应,实际只有5步需要简单后处理或纯化。微波加热提高了反应速率,反应时间明显缩短。合成总收率约15%,终产物HPLC纯度大于99.5%。结论此微波辅助新工艺试剂易得、操作简单、条件温和,可用于瑞博西尼的放量研究,亦适于2,6,7-取代吡咯并嘧啶类似物小分子库的快速建立。     

托法替尼中间体的合成工艺改进

以丙二腈为起始原料,与溴代乙醛缩二乙醇(2)发生α-烷基化反应,后经硫脲环合、脱保护、环合、酸化、脱除巯基、Sandmeyer氯代等反应,得到托法替尼中间体,4-氯-7H-吡咯并[2,3-d]嘧啶(1),总收率49%(以2计),纯度99.10%。该工艺有3点创新：(1)将巯基化合物成盐,直接从反应溶剂乙醇中析出,克服了巯基化合物难以纯化的技术难点,以较高收率(85%)制得4,6-二氨基-5-(2,2-二乙氧基乙基)嘧啶-2-硫醇钾盐(4);(2)一锅三步实现盐酸存在下的脱保护、环合、酸化反应,得到4-氨基-7H-吡咯[2,3-d]嘧啶-2-硫醇(5),简化了工序;(3)成功革除了三氯氧磷、2-甲基-3,3-二氯丙烯腈、2-氯乙醛等有毒试剂的使用,工艺更安全,尤其是三氯氧磷的革除大幅减少了三废,绿色环保。优化后的工艺原料经济易得、反应条件温和、操作简便、收率提高,适合工业化生产。     

7α,17α-二甲基-17β-羟基雄甾-4-烯-3酮的若干A环骈杂环衍生物的合成

将7α-甲基引进甲基睾丸素后,经甲酰化反应制得2-羟次甲基衍生物(Ⅳ),再先后转变为单肟(Ⅴ)、双肟(Ⅵ),环合得70,17α-二甲基-17β-羟基-雄甾-4-烯骈[2,3-c]呋咱(Ⅶ)。也可使(Ⅳ)与水合肼或盐酸羟胺作用,分别制得相应的雄甾-4-烯骈[3,2-c]吡唑(Ⅷ)及雄甾2,4-二烯骈[2,3-d]异(口恶)唑(Ⅸ)。在酸性反应条件下合成(Ⅸ)时,还分离出它的重排脱水产物7α,17α,17β-三甲基雄甾-2,4,13-三烯骈[2,3-d]异(口恶)唑(Ⅹ)。     

吡咯并嘧啶类化合物的合成及其抑制JAK3激酶活性的研究

目的设计合成吡咯并嘧啶类化合物并研究其抑制JAK3激酶的活性。方法以4-氯-7H-吡咯并[2,3-d]嘧啶为原料,经过取代、氨基脱保护和N-酰化反应合成两类(Ⅰa和Ⅰb)吡咯并嘧啶类化合物,经体外细胞试验测定其对JAK3激酶的抑制活性。结果设计并合成了8个新化合物,结构经1H-NMR和HR-MS确证。初步活性测试结果显示Ⅰa-1和Ⅰb-3对JAK3的抑制强度与阳性对照药tofacitinib相近。结论目标化合物对JAK3依赖的DAUDI细胞抑制活性较好,对非JAK3依赖的BT-20细胞抑制作用弱。     

2-取代-4-氨基噻吩并[3,2-d]嘧啶类化合物的合成及抗增殖活性

目的设计并合成2-取代-4-氨基噻吩并[3,2-d]嘧啶类化合物,评价其体外抗增殖活性。方法以3-氨基-2-噻吩甲酸甲酯为起始原料,经6步反应合成目标化合物;以CP-31398为阳性对照药,采用MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]法测定了目标化合物对肿瘤细胞株H-460和HT-29的抗增殖活性。结果与结论合成16个未见文献报道的化合物,其结构经1H-NMR、MS确证;5个化合物显示较好的抗增殖活性,其中,化合物8n活性突出,为CP-31398的4-5倍。     

疟疾防治药物的研究——Ⅶ.2,4-二哌啶(或吡咯啶)基-6-取代氨基喹唑啉衍生物的合成及其抗疟作用

本文报道2,4-二哌啶(或吡咯啶)基-6-取代氨基喹唑啉衍生物的合成及其抗疟作用。该类化合物经伯氏鼠疟原虫(Plasmodium berghei)抑制性治疗和约氏鼠疟原虫(P.yoelii)病因性预防筛选,发现有16个化合物对感染约氏鼠疟原虫的小鼠有病因性预防作用,其中以Ⅵ₁₁,Ⅷ₁,Ⅸ₂及Ⅸ₄四个化合物效果最好,每天口服2.5 mg/kg,连续三天,可使小鼠得到完全保护。此类化合物的合成,是以2,4-二氯-6-硝基喹唑啉与哌啶或吡咯啶缩合,经还原得2,4-二哌啶(或吡咯啶)基-6-氨基喹唑啉,然后与相应的取代苯甲醛缩合成席夫氏碱,再经还原和亚硝化而制得。     

N-酰基-N′-(2-吲哚甲酰基)肼的合成及其生理活性

2-吲哚甲酰肼与取代基为H,CH₃,ClCH₂,C₂H₅,CH₃(CH₂)₃,4-Br-C₆H₄和3-吡啶基的甲酰氯在DMF中反应时均得双酰肼化合物。但在乙腈中与乙酰氯或丙酰氯反应时,分别得到两种失水产物,经¹³C核磁共振仪分析表明其为吲哚2位(口恶)二唑取代的化合物,其余仍为双酰肼化合物。体外试验显示;N-甲酰基,N-氯代乙酰基-2-吲哚甲酰肼(Ⅰ,Ⅲ)和2-[2-(5-乙基)-1,3,4-(口恶)二唑]-吲哚(Ⅸ)对强毒人型结核菌H₃₇RV有抑制作用。     

Synthesis, cytotoxicity, and antiviral activity of some acyclic analogues of the pyrrolo[2,3-d]pyrimidine nucleoside antibiotics tubercidin, toyocamycin, and sangivamycin

A number of 7-[(1,3-dihydroxy-2-propoxy)methyl]pyrrolo[2,3d-d]pyrimidine derivatives that are structurally related to toyocamycin and sangivamycin and the seco nucleosides of tubercidin, toyocamycin, and sangivamycin were prepared and tested for their biological activity. Treatment of the sodium salt of 4-amino-6-bromo-5-cyanopyrrolo[2,3-d]-pyrimidine with 1,3-bis(benzyloxy)-2-propoxymethyl chloride afforded compound 3, which without isolation was debrominated to obtain 4-amino-5-cyano-7-[[1,3-bis(benzyloxy)-2- propoxy]methyl]pyrrolo[2,3-d]pyrimidine. Although catalytic hydrogenolysis failed, the benzyl ether functionalities of 4 were successfully cleaved by boron trichloride to afford 4-amino-5-cyano-7-[(1,3-dihydroxy-2- propoxy)methyl]pyrrolo[2,3-d]pyrimidine. Conventional functional group transformation of the cyano group of 6 provided a number of novel 5-substituted derivatives. Tubercidin (8a), toyocamycin (8b), and sangivamycin (8c) were treated separately with sodium metaperiodate and then with sodium borohydride to afford the 2',3'-seco derivatives 9a-c, respectively. The acyclic nucleoside 4-chloro-2-(methylthio)-7-[[1,3-bis(benzyloxy)-2- propoxy]methyl]pyrrolo[2,3-d]pyrimidine was aminated, desulfurized with Raney Ni, and then debenzylated to provide the tubercidin analogue 11. Cytotoxicity evaluation against L1210 murine leukemic cells in vitro showed that although the parent compounds tubercidin (8a), toyocamycin (8b), and sangivamycin (8c) were very potent growth inhibitors, the acyclic derivatives 6, 7a-c, and 9a-c had only slight growth-inhibitory activity. Evaluation of compounds 6, 7a, 7b, 7c, 9a, 9b, 9c, 11 for cytoxicity and activity against human cytomegalovirus (HCMV) and herpes simplex virus type 1 (HSV-1) revealed that only the carboxamide (7a) and the thioamide (7c) were active. Compound 7c was the more potent of the two, inhibiting HCMV but not HSV-1 at concentrations producing little cytotoxicity.     

Antiviral activity of C-5 substituted tubercidin analogues

The pyrrolo[2,3-d]pyrimidine nucleoside antibiotics tubercidin, toyocamycin, and sangivamycin and the synthetic analogues 5-chloro-, 5,6-dichloro-, 5-bromo-, 6-bromo-, 5,6-dibromo-, 5-iodo-, 5-(1-hydroxyethyl)-, 5-(1-methoxyethyl)-, (E)-5-(2-bromoethenyl)-, (E)-5-(2-cyanoethenyl)-, 5-(2-buten-1-yl)-, 5-(3-hydroxypropyl)-, and 5-butyltubercidin were evaluated for their antiviral properties against six RNA viruses and three DNA viruses in HeLa cell, primary rabbit kidney cell, and Vero cell cultures. Most of the derivatives had substantial activity against the RNA viruses, with the least activity shown by 6-bromo-, 5,6-dichloro-, and 5,6-dibromotubercidin. The C-5 substituted derivatives were quite toxic for the host cells. 5-(1-Hydroxyethyl)-, 5-(1-methoxyethyl)-, and 5-(2-buten-1-yl)tubercidin were more selective against reovirus type 1, parainfluenza virus type 3 and Coxsackie virus B4 than tubercidin and the 5-halotubercidins. When tested for in vivo activity against Coxsackie B4 virus infection in newborn NMRI mice, 5-(1-hydroxyethyl)- and 5-(1-methoxyethyl)tubercidin caused a significant decrease in the mortality rate at a dose level of 100 micrograms per mouse. The inhibitory effects on L-1210 cell growth were also determined, and toyocamycin (ID50 = 0.006 micrograms/mL) was found to be the most active compound. This study demonstrates the significance of structural modification at C-5 and the potential of C-5 substituted analogues of tubercidin as biologically active agents.     

Synthesis and antiviral activity of certain 4- and 4,5-disubstituted 7-[(2-hydroxyethoxy)methyl]pyrrolo[2,3-d]pyrimidines

In vitro evaluation of a series of previously prepared tubercidin analogues revealed that certain 5-halogen-substituted analogues were active against human cytomegalovirus (HCMV) at concentrations lower than those that produced comparable cytotoxicity in uninfected cells. In contrast, tubercidin was cytotoxic at all antiviral concentrations. Even though the antiviral selectivity of the 5-substituted compounds was slight, this observation led us to prepare a series of acyclic analogues. Treatment of the sodium salt of 4-chloropyrrolo[2,3-d]pyrimidine (2) with (2-acetoxyethoxy)methyl bromide (2a) provided the acyclic nucleoside 4-chloro-7-[(2-acetoxyethoxy)methyl]pyrrolo[2,3-d]pyrimidine (3). A nucleophilic displacement of the 4-chloro group with methoxide, methylamine, and dimethylamine yielded the corresponding 4-substituted compounds 4, 5, and 6, respectively, in good yield. Electrophilic substitution (chlorination, bromination, and iodination) was effected at the C-5 position of compound 3 with N-chlorosuccinimide, N-bromosuccinimide, and iodine monochloride, respectively, in methylene chloride. Removal of the acetyl group from these intermediates (7a-9a) with methanolic ammonia at room temperature afforded the 5-chloro (7b), 5-bromo (8b), and 5-iodo (9b) derivatives of 4-chloro-7-[(2-hydroxyethoxy)methyl]pyrrolo[2,3-d]pyrimidine. Treatment of compounds 7b-9b with methanolic ammonia at an elevated temperature produced the corresponding 5-halotubercidin analogues 10, 11, and 12, respectively. An alternate procedure for the preparation of these 4,5-disubstituted 7-[(2-hydroxyethoxy)methyl]pyrrolo[2,3-d]pyrimidines involved an electrophilic substitution prior to the condensation of the heterocycle with 2a. Treatment of 2 with N-chlorosuccinimide and N-bromosuccinimide gave compounds 13a and 13b, respectively. The condensation of 13a and 13b with 2a and subsequent treatment with methylamine and ethylamine furnished the corresponding 5-halo-4-substituted-pyrrolo[2,3-d]pyrimidines 14a, 14b, 14c, and 14d, respectively. Evaluation of the target compounds (4-6, 7b-9b, 10-12, and 14a-14d) for cytotoxicity and activity against HCMV and herpes simplex virus type 1 (HSV-1) revealed that all compounds except the 5-halogen-substituted compounds 10, 11, and 12 were inactive. Compounds 10, 11, and 12 were active against both viruses at noncytotoxic concentrations. The activity of compound 11 was particularly noteworthy, being at least 10-fold more potent than acyclovir.     

Synthesis and antiviral activity of certain 4-substituted and 2,4-disubstituted 7-[(2-hydroxyethoxy)methyl]pyrrolo[2,3-d]pyrimidines

Treatment of the sodium salt of 4-chloro-2-(methylthio)pyrrolo[2,3-d]pyrimidine (2) with (2-acetoxyethoxy)methyl bromide (3) has provided 4-chloro-2-(methylthio)-7[(2-acetoxyethoxy)methyl]pyrrolo[2,3- d]pyrimidine (4). Ammonolysis of 4 at room temperature gave 4-chloro-2-(methylthio)-7-[(2-hydroxyethoxy)methyl]pyrrolo[2,3- d]pyrimidine (5). However, ammonolysis of 5 at 130 degrees C furnished 4-amino-2-(methylthio)-7-[(2-hydroxyethoxy)methyl]-pyrrolo[2,3- d]pyrimidine (6), which on desulfurization with Raney Ni yielded 4-amino-7-[(2-hydroxyethoxy)-methyl]pyrrolo[2,3-d]pyrimidine (7) (acyclic analogue of tubercidin). The oxidation of 6 with m-chloroperbenzoic acid provided the sulfone derivative 8. A nucleophilic displacement of the 2-methylsulfonyl group from 8 with methoxide anion provided 4-amino-2-methoxy-7-[(2-hydroxyethoxy)methyl]pyrrolo[2,3-d]pyrimidine (9). Demethylation of 9 with iodotrimethylsilane gave 4-amino-2-hydroxy-7-[(2-hydroxyethoxy)methyl]pyrrolo[2,3-d]pyrimidine (10). Treatment of 2,4-dichloropyrrolo[2,3-d]pyrimidine (11) with 3 gave the protected acyclic compound 12, which on deacetylation and ammonolysis under controlled reaction conditions gave 2,4-dichloro-7-[(2-hydroxyethoxy)-methyl]pyrrolo[2,3-d]pyrimidine (13) and 4-amino-2-chloro-7-[(2-hydroxyethoxy)methyl]pyrrolo[2,3- d]pyrimidine (14), respectively. The condensation of 2-acetamido-4-chloropyrrolo[2,3-d]pyrimidine (15) with 3 gave the protected acyclic compound 16, which on concomitant deacetylation and ammonolysis with methanolic ammonia at an elevated temperature yielded 2,4-diamino-7-[(2-hydroxyethoxy)methyl]pyrrolo[2,3-d]pyrimidine (17) in moderate yield. In tests involving human cytomegalovirus (HCMV) and herpes simplex virus type 1 (HSV-1), only slight activity and cytotoxicity were observed. The most active compounds (12 and 13) were slightly more active against HCMV than acyclovir, but both compounds were inactive against HSV-1. The activity against HCMV, however, was not well separated from cytotoxicity leading to the conclusion that these compounds did not merit further study.     

3,4-二氢-海南新碱类似物的合成及抗溃疡作用

为考察3,4-二氢-海南新碱类似物C₁取代基对化合物抗溃疡活性的影响,设计并合成了13对共26个新的3,4-二氢-海南新碱类似物,A,B互为非对映异构体,大部分化合物在大鼠冷应激溃疡模型中有一定的抗溃疡活性,其中IX₃A,IX₇A,IX₈A,IX₁₂A,IX₁₂B,IX₁₃A6个化合物表现较强活性,超过西咪替丁。对构效关系进行了初步分析。     

Synthesis, antiproliferative, and antiviral activity of certain 4-aminopyrrolo[2,3-d]pyridazine nucleosides: an entry into a novel series of adenosine analogues.

The preparation of novel heterocyclic base modified adenosine analogues, the 4-aminopyrrolo[2,3-d]pyridazine nucleosides, is described. Crucial to their successful preparation was the use of the pyrrole glycoside intermediates 3-cyano-2-formyl-1-(2,3,5-tri-O-benzyl-beta-D-ribofuranosyl)pyrrole (11) and 3-cyano-2-formyl-1-(2,3,5-tri-O-benzyl-beta-D-arabinofuranosyl)pyrrole (17). Treatment of 11 and 17 with hydrazine dihydrochloride followed by treatment with boron trichloride provided 4-amino-1-beta-D-ribofuranosylpyrrolo[2,3-d]pyridazine (2) and 4-amino-1-beta-D-arabinofuranosylpyrrolo[2,3-d]pyridazine (3), respectively. 4-Amino-3-bromo-1-beta-D-ribofuranosylpyrrolo[2,3-d]-pyridazine (4) was prepared by a bromination of 4-amino-1-(2,3,5-tri-O-benzyl-beta-D-ribofuranosyl)pyrrolo[2,3-d]pyri daz ine (12) and subsequent removal of the benzyl groups with boron trichloride. Compounds 2-4 were evaluated for antiproliferative and antiviral activity. The tubercidin analogue (2) and its arabinosyl derivative (3) were virtually inactive in all assays. In contrast, the 3-bromo analogue 4 inhibited growth of L1210 and H. Ep. 2 cells. Compound 4 was also active against human cytomegalovirus and herpes simplex virus type 1, but the antiviral activity was not completely separated from cytotoxicity.     

白花丹参化学成分的研究

从白花丹参(Salvia miltiorrhiza f.alba)中分离出14个二萜醌类化合物,其中2个为新化合物,由光谱分析推定为1,2,15,16-四氢丹参醌(Ⅰ)和丹参醛(Ⅱ);另一个化合物Ⅲ与R₀-090680的结构相同。化合物Ⅰ和Ⅲ对淋巴白血病细胞P₃₈₈均有较强的抑制作用。     

Synthesis and antimicrobial activity of new substituted thienopyrimidines, their tetrazolyl and sugar derivatives

A series of substituted 5,6,7,8-tetrahydro-3H-benzo[4,5]thieno[2,3-d]pyrimidine-4-one derivatives were newly synthesized starting from 5,6,7,8-tetrahydro-3H-benzo[4,5]thieno[2,3-d]pyrimidine-4-one derivatives (1). Furthermore, their derived tetrazolyl as well as the N-substituted derivatives were also prepared. The antimicrobial activity of the prepared compounds against Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Aspergillus niger and Candida albicans were evaluated. The substituted thienopyrimidine derivatives 4 and 6 as well as the arylidine 10 were the highly active compounds.     

Synthesis and properties of the derivatives of 2-alkylthio-4-oxo-3,4- (and -1,4)-dihydropyrido-[2,3-d]pyrimidine-5- and -6-carboxylic acids

Condensation of diethyl 2-amino-6-methylpyridine-3,4-dicarboxylate (I) with the corresponding isothiocyanates afforded derivatives of ethyl 4-oxo-2-thioxo-1,2,3,4-tetrahydropyrido [2,3-d]pyrimidine-5-carboxylate (V-VII). Alkylation of (V), (VI) and (XI) gave the corresponding derivatives of ethyl 2-alkylthio-4-oxo-3,4-(and 1,4)-dihydropyrido[2,3-d]pyrimidine-5- and -6- carboxylate [(XII-XVI), (XX-XXII)]. Some of the obtained compounds were active pharmacologically.     

A new biological role of sangivamycin; inhibition of protein kinases

During the screening for the inhibitors of protein kinase C (PKC), we found that a streptomycete produced an inhibitor in our bleb-forming assay (Osada et al., J. Antibiotics 41: 925, 1988). The inhibitor was isolated and identified as sangivamycin (4-amino-5-carboxamide-7-(D-ribofuranosyl)pyrrolo[2,3-d]pyrimidine). Biological activity of sangivamycin was compared with that of other 7-deazaadenosine group antibiotics, tubercidin and toyocamycin. Sangivamycin showed a strong inhibitory activity against bleb-formation of K562 cells and PKC. On the other hand, tubercidin and toyocamycin had only weak activities in both assays. This paper deals with a new biological activity of sangivamycin, that of an inhibitor of protein kinases, especially PKC.