Identification of phase-I metabolites and chronic toxicity study of the Kv1.3 blocker PAP-1 (5-(4-phenoxybutoxy)psoralen) in the rat

1. PAP-1 (5-(4-phenoxybutoxy)psoralen), a potent small-molecule blocker of the voltage-gated potassium Kv1.3 channel, is currently in preclinical development for psoriasis. This study was undertaken to identify the major phase I metabolites of PAP-1 in Sprague-Dawley (SD) rats. 2. Five phase I metabolites, that is 5-(oxybutyric-acid)psoralen (M1), 5-[4-(4-hydroxybutoxy)]psoralen (M2), 5-[4-(4-hydroxyphenoxy)butoxy]psoralen (M3), 5-[4-(3-hydroxyphenoxy)butoxy]psoralen (M4), and 8-hydroxyl-5-(4-phenoxybutoxy)psoralen (M5), were isolated from the bile of rats and identified by mass spectrometry and NMR spectroscopy. The last four metabolites are new compounds. 3. Incubation of PAP-1 with SD rat liver microsomes rendered the same five major metabolites in a nicotinamide adenine dinucleotide phosphate (NADPH)-dependent manner suggesting that cytochrome P450 (CYP) enzymes are involved in PAP-1 metabolism. Inhibitors of rat CYP1A1/2 (alpha-naphthoflavone) and CYP3A (ketoconazole) but not CYP2D6 (quinidine), CYP2E (diethyldithiocarbamate), or CYP2C9 (sulphaphenazole) blocked the metabolism of PAP-1 in rat microsomes. 4. Of the five metabolites M3, M4, and M5 were found to inhibit Kv1.3 currents with nanomolar IC50s, while M1 and M2 were inactive. Our results identified the Kv1.3-inactive M1 as the major phase I metabolite, and suggest that hydroxylation and O-dealkylation are the major pathways of PAP-1 metabolism. 5. We further conducted a 6-month repeat-dose toxicity study with PAP-1 at 50 mg/kg in both male and female Lewis rats and did not observe any toxic effects.     

HPLC-UV法测定皮肤给药后大鼠血浆5-（4-苯氧丁氧基）补骨脂素浓度

目的：建立大鼠血浆中5-（4-苯氧丁氧基）补骨脂素[5-（4-phenoxybutoxy）psoralen,PAP-1]高效液相色谱-紫外（HPLC-UV）测定方法,并用于大鼠皮肤给药后血浆PAP-1浓度的测定。方法：以异欧前胡素为内标,血浆样品经乙酸乙酯提取;应用Ultimate Column AQ-C18色谱柱（150 mm×4.6 mm,5μm）;流动相为甲醇-水（75：25）,流速0.8 mL/min,检测波长311 nm;采用PAP-1与内标峰面积比值进行定量。大鼠皮肤给药PAP-1,检测不同时间血药浓度。结果：本方法最低检测限为5.3μg/L（S/N〉10）,线性范围为5.3～2120μg/L,血浆中PAP-1的日内和日间精密度为2.3%～6.0%,准确度为102%～105%,回收率为83.1%～86.5%。大鼠15 mg/kg皮肤给药后,PAP-1在4 h达峰。雄性和雌性大鼠的AUC分别为256和1209μg.L-1.h,Cmax分别为74和225μg/L。结论：本方法专属性强、简便高效,可用于测定药物代谢动力学实验中PAP-1的浓度。本文首次报道大鼠PAP-1皮肤给药的药物动力学过程,药代参数存在明显的性别差异。     

维生素B1催化合成1-(3-甲氧基-4-丙氧基-5-硝基苯基)-4-(3,4,5-三甲氧基苯基)-1,4-二酮

研究以NaCN和维生素B1为催化剂合成MK-287的关键中间体1-(3-甲氧基-4-丙氧基-5-硝基苯基)-4-(3，4，5-三甲氧基苯基)-1，4-二酮（1）的催化效果。结果表明：维生素B1较文献报道的催化剂ETB具有反应时间短、成本低廉的优点，可代替ETB合成目的化合物。     

Pharmacokinetics and tissue distribution of 3-((5-(6-methylpyridin-2-yl)-4-(quinoxalin-6-yl)-1H-imidazol-2-yl)methyl)benzamide; a novel ALK5 inhibitor and a potential anti-fibrosis drug

The authors investigated the pharmacokinetics and metabolism of 3-((5-(6-methylpyridin-2-yl)-4-(quinoxalin-6-yl)-1H-imidazol-2-yl)methyl)benzamide (IN-1130), a novel ALK5 inhibitor, which suppresses renal and hepatic fibrosis, and also exerts anti-metastatic effects on breast cancer-bearing MMTV-cNeu mice model. Plasma half-lives of orally administered IN-1130 were 62.6 min in mice, 76.6?±?10.6 min in dogs, 156.1?±?19.3 min in rats, and 159.9?±?59.9 min in monkeys. IN-1130 showed a high apparent permeability coefficient (P_app) of (45.0?±?2.3)?×?10^?6 cm s^?1 in in vitro permeability tests in a Caco-2 cell monolayer model. The bioavailability of orally administered IN-1130 was 84.9% in dogs and 34.4% in monkeys (oral dose, 5.5 mg kg^?1), 11.4% in rats and 8.95% in mice (oral dose, 50.3 mg kg^?1), respectively. Orally given IN-1130 was readily distributed into liver, kidneys and lungs. The major metabolite of IN-1130 (M1) was detected in the systemic circulation of rat and mouse and was purified and tentatively identified as 3-((4-(3-hydroxyquinoxaline-6-yl)-5-(6-methylpyridine-2-yl)-1H-imidazol-2-yl)methyl)benzamide or 3-((4-(2-hydroxyquinoxalin-6-yl)-5-(6-methylpyridine-2-yl)-1H-imidazol-2-yl)methyl)benzamide. The highest levels of M1 were found in liver. The results of this study suggest that IN-1130 has the potential to serve as an effective oral anti-fibrotic drug.     

LC-MS/MS法研究1-[1-(6-甲氧基-2-萘基)乙基]-2-(4-硝基苄基)-6,7-二甲氧基-1,2,3,4-四氢异喹啉氢溴酸盐在大鼠体内的代谢产物

采用液相色谱-串联质谱法对大鼠灌胃1-［1-(6-甲氧基-2-萘基)乙基］-2-(4-硝基苄基)-6,7-二甲氧基-1,2,3,4-四氢异喹啉氢溴酸盐(编号P91024)后粪便、尿液、胆汁和血浆中的主要代谢产物进行研究。通过比较给药样品和空白样品的全扫描总离子流色谱和选择离子扫描色谱图差别寻找I相代谢产物；根据其一级和二级质谱图，确定I相代谢产物的分子结构。完全提取I相代谢产物后的样品溶液，再用葡糖醛酸酶酶解，得II相结合物的苷元部分，采用与I相代谢产物鉴定同样方法寻找和鉴定II相代谢产物苷元的结构，进而确证II相代谢产物的分子结构。从大鼠粪便中鉴定出P91024的2个I相代谢物，从胆汁中鉴定出1个I相和5个II相代谢产物，从尿液中鉴定出1个I相和3个II相代谢产物，从血浆中鉴定出4个I相和1个II相代谢产物；并分别分析推测出它们的结构。P91024在大鼠体内被代谢转化为多种产物，利用LC-MS/MS可以快速寻找和鉴定。     

Synthesis and antileukemic activity of new 3-(1-phenyl-3-methylpyrazol-5-yl)-2-styrylquinazolin-4(3H)-ones

3-(1-Phenyl-3-methylpyrazol-5-yl)-2-styrylquinazolin-4(3H)-ones 14a-q and 15a-q were synthesized by refluxing in acetic acid the corresponding 2-methylquinazolinones 12 and 13 with the opportune benzoic aldehyde for 12 h. The synthesized styrylquinazolinones 14a-q and 15a-q were tested in vitro for their antileukemic activity against L1210 (murine leukemia), K562 (human chronic myelogenous leukemia) and HL60 (human leukemia) cell lines showing in some cases good activity.     

1-(4-溴苄基)-1-(4-溴苄氧基)脲药代动力学和体外代谢的研究

目的研究1-(4-溴苄基)-1-(4-溴苄氧基)脲(HY-D11)在大鼠体内药代动力学过程及其在大鼠肝脏中的代谢途径。方法建立HY-D11的UV-HPLC生物样本检测方法,灌胃给予HY-D11 66.7 mg·kg-1后,观察HY-D11在大鼠体内的药代动力学过程。研究CYP1A抑制剂(α-萘黄酮)、CYP2B抑制剂(舍曲林)、CYP2C抑制剂(槲皮素)、CYP2D抑制剂(育亨宾)、CYP2E抑制剂(双硫仑)及CYP3A抑制剂(醋竹桃霉素)对HY-D11在大鼠肝微粒体温孵液中代谢的影响。结果 HY-D11在大鼠体内的药代动力学过程符合二室模型。药代动力学参数t max为(3.67±0.52)h,C max为(1.61±0.13)mg·L-1,t1/2z为(3.41±3.91)h,平均滞留时间MRT0-∞为(7.05±0.29)h。HY-D11在大鼠微粒体温孵液中呈现明显的酶促动力学特征。醋竹桃霉素组与对照组相比,对HY-D11代谢有明显抑制作用(P<0.01);而与对照组相比,萘黄酮组、舍曲林组、槲皮素组、育亨宾组、双硫仑组对HY-D11的代谢均无抑制作用(P>0.05)。结论 HYD11在大鼠体内吸收、代谢速度较快,在大鼠体内代谢可能主要由CYP3A介导。     

1-(5-硝基吲哚-2-羰基)-4-[3-(1-甲基乙胺基)-2-吡啶基]哌嗪的制备

地拉韦定(delavirdine)是由美国Pharmacia&Upjohn公司研制的非核苷HIV-1逆转录酶抑制剂(NNRTIs),临床与其它抗HIV药联合使用,用于治疗获得性免疫缺陷综合征[1].1-(5-硝基吲哚-2-羰基)-4-[3-(1-甲基乙胺基)-2-吡啶基]哌嗪(1)是其重要中间体.文献[2-4]以5-硝基吲哚-2-羧酸(2)和1-[3-(1-甲基乙胺基)-2-吡啶基]哌嗪(3)为原料,在1-(3-二甲胺基丙基)-3-乙基碳二亚胺(EDC)作用下缩合制得.EDC价昂,后处理用氯仿作提取溶剂,且需通过柱色谱纯化,不适于规模化生产.     

Synthesis and cytotoxic activity of 1-(1-benzoylindoline-5-sulfonyl)-4-phenylimidazolidinones

The novel 1-(1-benzoylindoline-5-sulfonyl)-4-phenyl-4,5-dihydroimidazolones 2 shows highly potent and broad cytotoxicities. Their cytotoxicities against human lung carcinoma A549, human chronic myelogenous leukemia K562, and human ovarian adenocarcinoma SK-OV-3 are compatible with doxorubicin. Compound 2p (1-[(4-aminobenzoyl)indoline-5-sulfonyl])-4-phenyl-4,5-dihydroimidazolone) exhibits a cytotoxicity that is far more potent than doxorubicin and also exhibits highly effective antitumour activities against murine (3LL, Colon 26) and human xenograft (NCI-H23, SW620) tumor models.     

6-(1-溴乙基)-4-氯-5-氟嘧啶合成新方法

以氟乙酸乙酯为起始原料,改进抗真菌药物伏立康唑中间体6-(1-溴乙基)-4-氯-5-氟嘧啶的合成方法.该方法简化了生产工艺,降低了反应成本,反应条件温和,适合工业化生产,总收率为41.7%.     

Synthesis and Antimicrobial Activity of Some New 2-(3-(4-Aryl)-1-phenyl-1H-pyrazol-4-yl) Chroman-4-ones

Seven new 2-(3-(4-aryl)-1-phenyl-1H-pyrazol-4-yl) chroman-4-ones (4a-4g) have been synthesized by cyclization of 2-hydroxychalcone analogues of pyrazole 3a-3g using conc. HCl in acetic acid. The structures of the compounds 4a-4g were established by the combined use of (1)HNMR, IR and mass spectra. All the seven compounds were tested in vitro for their antibacterial activity against two Gram positive bacteria namely Staphylococcus aureus and Bacillus subtilis and two Gram negative bacteria Escherichia coli and Pseudomonas aeruginosa. The compounds 4b, 4c, 4e, 4f, 4g have displayed good antibacterial activity when compared with commercially available antibiotic, ciprofloxacin. These compounds also were screened for their antifungal activity against two ear pathogenic fungi, namely Aspergillus Niger and A. flavus. The compounds 4a, 4c, 4d, 4g exhibited good antifungal activity when compared with commercially available antifungal, fluconazole.     

Deacetylation of 4-(5-acetylamino-2-furyl)thiazole and formation of 1-(4-thiazolyl)-3-cyano-1-propanone by rat liver tissues

The reductive metabolism of the rat carcinogen 4-(5-nitro-2furyl)thiazole (NFT) to 1-4-thiazolyl)-3-cyano-1-propanone (TCP) is reported. Formation of TCP from NFT involved furan ring fission. This could have occurred through involvement of either aminofuran or N-hydroxylaminofuran as precursors. To examine if 4-(5-amino-2-furyl)thiazole is a precursor for TCP, a stable model compound, 4-(5-acetylamino-2-furyl)thiazole (AAFT), was prepared and subjected to enzymatic deacetylation, using rat liver tissue homogenates. AAFT was synthesized by catalytic hydrogenation of NFT with 5% palladium on activated carbon, followed by acetylation with acetic anhydride. AAFT, a white crystalline powder, melted at 168–170°, had an extinction coefficient of 17.9 mM^?1 cm^?1 at 293 nm in ethyl acetate, and exhibited spectroscopic and mass spectral characteristics consistent with the assigned structure. Incubation with rat liver 10,000 g supernatant preparations resulted in the biotransformation of AAFT as evidenced by a decrease in absorption at 290 nm. Incubation of ¹⁴C-labeled AAFT followed by extraction with chloroform-diethyl ether (1:1) resulted in the recovery of a major portion (56%) of the radioactivity in the organic phase when the label was at the 2-position of the thiazole ring, while the major amount (82%) of radioactivity was recovered in the aqueous phase when the 1-¹⁴C-acetyl group was labeled. The radioactivity from the aqueous phase was extractable into the organic phase following acidification to pH 1, an observation consistent with deacetylation. Furthermore, the deacetylation product exhibited a mass spectrum, and retention times in gas and high pressure liquid chromatography, similar to those of synthetic TCP. These data establish 4-(5-amino-2-furyl)thiazole, derived from AAFT by deacetylation, as a precursor for TCP.     

4-(4-苯基哌嗪-1-基)-3-O-甲基-D-肌醇的制备及镇咳活性

右旋肌醇甲醚经缩酮制化得3-O-甲基-1，2：5，6-二-O-异亚丙基-D-肌醇，经甲磺酰化、水解、与4-苯基哌嗪缩合制得左羟丙哌嗪类似物4-（4-苯基哌嗪-1-基）-3-O-甲基-D-肌醇，总收率38％。镇咳药效实验表明，目标物对致咳小鼠的镇咳作用明显，但不及左羟丙哌嗪。     

3-(5-甲氧基-1,5-二氧代戊基)-(4S)-苯基噁唑烷-2-酮的制备

依折麦布(ezetimibe),化学名为(3R,4S)-1-(4-氟苯基)-3-[(3S)-3-(4-氟苯基)-3-羟丙基]-4-(4-羟苯基)-2-氮杂环丁酮,是由先灵葆雅和默克公司联合研发的新型胆固醇吸收抑制剂,2002年11月首次在德国上市,2007年8月在我国上市,临床主要用于治疗高胆固醇血症~[1].3-(5-甲氧基-1,5-二氧代戊基)-(4S)-苯基噁唑烷-2-酮(1)是合成依折麦布的重要中间体.     

4-叔丁基-5-(12,4,-三唑-1-基)-2-苄亚氨基噻唑的体外抗癌活性研究

目的研究4-叔丁基-5-(1,2,4-三唑-1-基)-2-苄亚氨基噻唑类衍生物对宫颈癌细胞系Hela、肝癌细胞系Bel 7402、鼻咽癌细胞系CNE 2的体外抗癌活性。方法细胞抑制率采用四甲基偶氮唑蓝(MTT)法测定。每种试样设置5个浓度梯度(0.025、0.05、0.10、.25、0.5μmol/ml),每个浓度取4个平行样,并设置无药对照实验,用半数抑制浓度(IC50)评价目标化合物的抗癌活性。结果被测化合物对3种癌细胞均表现出一定的抑制作用,其中化合物5对Hela癌细胞和Bel 7402癌细胞的IC50值分别为0.076 1、0.062 8μmol/ml;化合物6对CNE 2癌细胞的IC50值为0.050 8μmol/ml。结论生物活性测试表明该类型氨基噻唑衍生物具有较好的抗癌活性,值得进一步研究和关注。     

5-Substituted pyrimidine acyclic nucleoside analogues 1-cyanomethyl- and 1-(4-cyanobutyl)-5-substituted uracils as candidate antitumor agents

A number of 5-substituted pyrimidine acyclic nucleosides were synthesized and tested for in vitro cytotoxicity against four cell lines (J-82 cell, P-388 cell, FM-3A cell and U-938 cell lines). Synthesis of 1-cyanomethyl-5-substituted pyrimidines (1a-e) and 1-(4-cyanobutyl)-5-substituted pyrimidines (2a-e) was accomplished from the series of alkylation reactions of 5-substituted uracils with the corresponding chloroacetonitrile and 5-chlorovaleronitile in DMSO under 50 degrees C temperature. These 5-substituted pyrimidine acylic nucleosides (1a-e and 2a-e) exhibited moderate to significant activity against four cell lines.     

N-(双膦羧次甲基)-6-(5-氟-2,4-二氧代-3,4-二氢-2H-嘧啶-1-基)-6-氧代-己酰胺的合成及其初步的骨靶向性研究

目的合成N-（双膦羧次甲基）-6（5-氟-2，4-二氧代3，4-二氢-2H-嘧啶-1-基）-6-氧代-己酰胺，并进行初步体外骨靶向性实验。方法以5-氟尿嘧啶为原料，经硅烷化、缩合、氢解3步合成6-（5-氟-2，4-二氧代-3，4-二氢-2H-嘧啶-1-基）-6-氧代-己酸（2），用二氯亚砜氯化后再与含氨基的偕二膦酸酯偶联，最后再用溴代三甲基硅烷特异性解离掉膦酸酯得到目标化合物L，并采用羟磷灰石晶体吸附实验考察目标物的骨靶向性。结果合成了目标物L，并利用^-1H—NMR、IR和MS进行了结构确证。结论体外骨靶向性实验结果显示目标物L有较好的骨靶向性。     

Synthesis of 4-(2,4-dioxo-5-pyrimidyl)-1,4-dihydropyridine derivatives

Hantzsch synthesis of 5-formyluracil (1), methyl acetoacetate (2) and methyl 3-aminocrotonate (3) gave 2,6-dimethyl-4-(2,4-dioxo-5-pyrimidyl)-1, 4-dihydropyridine-3, 5-dicarboxylic acid dimethylester (4a) in 54.6% yield. As the same procedure, 1,3-dimethyl-5-formyl-uracil (6) gave 2,6-dimethyl-4-(1,3-dimethyl-2,4-dioxo-5-pyrimidyl)-1, 4-dihydropyridine-3,5-dicarboxylic acid dimethyl ester (7a) in 52.2% yield.4a was methylated to afford7a also in 52% yield.     

血吸虫病化学治疗的研究 ⅩⅩⅩⅥ.4-苯基(或烯丙基)-5-(吡嗪-2)-1,2,4-三唑-3-硫酮衍生物的合成

本文报导28个4-苯基(或烯丙基)-5-(吡嗪-2)-1,2,4-三唑-3-硫酮衍生物的合成。这类化合物的合成是以吡嗪甲酸乙酯与水合肼反应得2-吡嗪甲酰肼,再与不同的异硫氰酸酯作用后,在2N氢氧化钠溶液中环合而得Ⅱ或Ⅲ,然后经烷化、酰化及Mannich反应,分别制得相应的化合物。其中Ⅲ₁和Ⅲ₂对感染日本血吸虫小白鼠有明显肝移作用。     

Synthesis and antifungal activity of new N-(1-phenyl-4-carbetoxypyrazol-5-yl)-, N-(indazol-3-yl)- and N-(indazol-5-yl)-2-iodobenzamides

N-(1-Phenyl-4-carbetoxypyrazol-5-yl)-, N-(indazol-3-yl)- and N-(indazol-5-yl)-2-iodobenzamides 6, with a Benodanil-like structure, were synthesized by refluxing in acetic acid the corresponding benzotriazinones 5 with potassium iodide for 1 h in order to study the role on the antifungal activity of the N-substitution with an aromatic heterocyclic system on benzamide moiety. Among the tested iododerivatives, compounds 6d,f,g,h possess interesting activities toward some phytopathogenic fungal strains.