alpha-Aminophosphonic acids and their derivates, as phosphorous analogs of amino acid, have attracted much attention because they show wide biological activities. In this paper, the title compounds, {phenyl-[(pyridine-4-carbonyl)amino]methyl}phosphonic acid dialkyl esters, were synthesized via Mannich reaction and peptide coupling reaction. Their structures were confirmed by elemental analysis, IR, H-1 NMR, C-13 NMR and MS spectra, and the antibacterial and antitumor activities of these compounds were given in this paper

FANG, Hua

FANG, Mei-Juan

LIU, Xiao-Xia

XU, Peng-Xiang

ZHAO, Yu-Fen

方华

方美娟

留筱厦

许鹏翔

赵玉芬

Chinese Journal of Organic Chemistry

Xiamen University Institutional Repository

 2005年第 25卷 有 机 化 学 Vol. 25, 2005 第 4期, 466～469 Chinese Journal of Organic Chemistry No. 4, 466～469   * E-mail: yfzhao@xmu.edu.cn; Fax: 0592-2185780; Tel.: 0592-2185780. Received June 7, 2004; revised October 10, 2004; accepted November 10, 2004. 福建省重点基金(No. 2001F008)资助项目. ·研究简报· {苯基-[(吡啶-4-羰基)-氨基]-甲基}-膦酸二烷基酯的合成与波谱表征 方  华 a    方美娟 a    留筱厦 a    许鹏翔 a    赵玉芬*,a,b (a厦门大学化学化工学院福建省化学生物学省重点实验室  厦门 361005) (b清华大学生命有机磷化学及化学生物学教育部重点实验室  北京 100084) 摘要  α-氨基膦酸及其衍生物作为天然氨基酸的含磷类似物, 具有多种生物活性, 它的合成与性质的研究引起人们的浓厚兴趣. 合成了一系列含磷化合物, 产物经元素分析、红外光谱、核磁共振、质谱鉴定, 并初步研究这些化合物的抗菌与抗肿瘤生物活性.  关键词  α-氨基膦酸; 合成; 波谱表征; 生物活性 Synthesis and Spectral Characterization of {Phenyl-[(pyridine-4- carbonyl)amino]methyl}phosphonic Acid Dialkyl Ester FANG, Huaa    FANG, Mei-Juana    LIU, Xiao-Xiaa    XU, Peng-Xianga ZHAO, Yu-Fen*,a,b (a Key Laboratory for Chemical Biology of Fujian Province, Department of Chemistry, Xiamen University, Xiamen 361005) (b Key Laboratory for Bioorganic Phosphorus Chemistry and Chemical Biology, Ministry of Education, Department of Chemistry, Tsinghua University, Beijing 100084) Abstract  α-Aminophosphonic acids and their derivates, as phosphorous analogs of amino acid, have at-tracted much attention because they show wide biological activities. In this paper, the title compounds, {phenyl-[(pyridine-4-carbonyl)amino]methyl}phosphonic acid dialkyl esters, were synthesized via Mannich reaction and peptide coupling reaction. Their structures were confirmed by elemental analysis, IR, 1H NMR, 13C NMR and MS spectra, and the antibacterial and antitumor activities of these compounds were given in this paper. Keywords  α-aminophosphonic acid; synthesis; spectral characterization; biological activityα-氨基膦酸与天然氨基酸具有结构上的类似性[1], 显示良好的生物活性如: 除草[2]、植物生长调节、杀菌、抗病毒及酶的抑制剂等[3]. 二烷基膦酸酯类化合物具有螯合结构, 有良好的配位性能, 易形成双核配合物, 可以用来研究电子在二烷基膦酸酯类物质中的转移能 力[4]. 对 α-氨基膦酸及其衍生物的研究, 主要是将 α-氨基膦酸的母体与其他有某种生物活性的结构单元相连, 合成出结构新颖的 α-氨基膦酸及其衍生物, 希望找到高 活性、低毒的农药和医药新品种[5]. 本论文采用稍加修改的 Ph3P 和 C2Cl6法[6,7]进行酰胺键的缩合(Scheme 1), 此法为一步反应, 中间体活化酯不需提纯, 反应几乎是定量. 它具有原料便宜易得、条件温和、操作方便、易提纯和收率高等特点[8], 是合成酰胺键的有效方法之一. 并发现所合成的含磷化合物都具有一定的抗菌抗肿瘤生物活性, 所以合成这类化合物就具有很重要的意义, 下面就这些化合物的具体合成做系统介绍.    No. 4 方华等：{苯基-[(吡啶-4-羰基)-氨基]-甲基}-膦酸二烷基酯的合成与波谱表征 467   R＝Me (a); Et (b); i-Pr (c) Scheme 1 1  实验 1.1  仪器与试剂 Yanaco 显微熔点仪, 温度计已加校正; 元素分析EA由 Flash EA 1112元素自动分析仪给出; 红外 IR由Nicolet AVATAR 360 FT-IR计记录, 溴化钾压片; 核磁1H NMR和 13C NMR Varian 500 Hz型仪器测定, D2O或CDCl3 为溶剂 , TMS 为内标 ; 质谱 MS 由 Bruker ESQSUIRE-3000 离子阱型质谱仪测定; M3550 型酶标仪(测量波长 570 nm, 参考波长 655 nm).  试剂与溶剂为化学纯或分析纯, 用前要进一步处理.  1.2  实验步骤 1.2.1  α-氨基苄基膦酸二烷基酯的盐酸盐(1a～1c)的制备 冰浴下三氯化磷与相应的醇反应得到相应的二烷基亚磷酸酯. α-氨基苄基膦酸二烷基酯的盐酸盐(1a～1c)根据文献的方法制备, 制备得到的产物 1a～1c 与文献报道一致[9～11]. 1.2.2  {苯基-[(吡啶-4-羰基)-氨基]-甲基}-膦酸二烷基酯的制备 冰浴下, 干燥过的 15 mL l,2-二氯乙烷中加入 Ph3P (3.93 g, 15 mmol)和 C2Cl6 (3.58 g, 15 mmol), 反应 1 h; 反应液慢慢加到 90 mL 溶有 α-吡啶酸(1.23 g, 10 mmol), α-氨基苄基膦酸二烷基酯的盐酸盐(10 mmol)和 Et3N (4 mL)的干燥1,2-二氯乙烷中, 反应24 h, 以上反应均在无水无氧的条件下进行. 反应溶液滤去三乙胺盐, 滤液旋干, 再加入 30 mL乙醇. 此溶液用 0.5 mol/L HCl酸化至pH 为 1～2, 乙醚洗涤两次除去有机相, 水相用 4% NaOH调 pH至 11, 二氯甲烷萃取产物, 浓缩, 干燥, 粗产物在乙酸乙酯和石油醚混合液里重结晶, 即得产物. {苯基-[(吡啶-4-羰基)-氨基]-甲基}-膦酸二甲酯 (2a): 白色晶体, 产率 76.5%, m.p. 134.1～135.5 ℃; 1H NMR (CDCl3) δ: 9.46～9.92 (m, NH), 8.58 (d, J＝5 Hz, 4H, Py), 7.20～7.60 (m, 5H, ArH), 5.42 (dd, JP-CH＝21 Hz, JNH-CH＝9 Hz, 1H, CH), 4.02 (s, 6H, 2OCH3); 13C NMR (CDCl3) δ: 149.44, 123.94, 145.22 (Py), 133.69, 128.90, 127.25, 128.73 (Ar), 168.28 (C＝O), 52.39 (CH), 55.55, 53.93 (2OCH3); IR (KBr) ν: 3402 (N—H), 1662 (C＝O), 1225 (P＝O), 1074 (P—O—C), 1557, 1511, 1495, 1453 (芳环骨架振动) cm－1; MS m/z (%): 210.9 (19.48), 321  (M＋H＋, 100), 641.0 (2M＋H＋, 1.01). Anal. calcd for C15H17N2O4P•H2O: C 53.26, H 5.66, N 8.28; found C 53.55, H 5.54, N 8.09. {苯基-[(吡啶-4-羰基)-氨基]-甲基}-膦酸二乙酯 (2b): 白色晶体, 产率 82.0%, m.p. 107.0～108.1 ℃; 1H NMR (CDCl3) δ: 7.86～8.37 (m, NH), 8.18 (d, J＝5 Hz, 4H, Py), 7.23～7.62 (m, 5H, ArH), 5.76 (dd, JCH-NH＝9 Hz, JP-CH＝21 Hz, 1H, CH), 3.98 (t, JCH2-CH3＝8 Hz, 4H, 2CH2- CH3), 1.22 (d, JCH2-CH3＝8 Hz, 6H, 2CH2CH3); 13C NMR (CDCl3) δ: 150.51, 121.19, 140.84 (Py), 134.64, 128.47, 128.17, 128.78 (Ar), 165.09 (C＝O), 51.40 (CH), 63.72, 63.20 (2CH2CH3), 16.44, 16.10 (2CH2CH3); IR (KBr) ν: 3440 (N—H), 1664 (C＝O), 1244 (P＝O), 1032 (P—O—C), 1596, 1546, 1500, 1456 (芳环骨架振动) cm－1; MS m/z (%): 211.0 (17.77), 348.9 (M＋H＋, 100), 696.7 (2M＋H＋, 10.16). Anal. calcd for C17H21N2O4P: C 58.62, H 6.08, N 8.04; found C 58.44, H 6.10, N 7.95. {苯基-[(吡啶-4-羰基)-氨基]-甲基}-膦酸二异丙基酯 (2c): 白色晶体, 产率 85.1%, m.p. 134.3～135.7 ℃; 1H NMR (CDCl3) δ: 7.58～8.26 (m, NH), 8.18 (d, J＝5 Hz, 4H, Py), 7.18～7.72 (m, 5H, ArH), 5.72 (dd, JP-CH＝21 Hz, JNH-CH＝9 Hz, 1H, CH), 4.50 [d, JCH-CH3＝6 Hz, 2H, 2CH(CH3)2], 1.20 [d, JCH-CH3＝6 Hz, 12H, 2CH(CH3)2]; 13C NMR (CDCl3) δ: 150.26, 121.42, 141.15 (Py), 135.13, 128.61, 128.15, 128.65 (Ar), 165.39 (C＝O), 51.27 (CH), 72.48, 71.96 [2CH(CH3)2], 24.12, 24.01, 23.86, 23.81 [2CH(CH3)2]; IR (KBr) ν: 3448 (N—H), 1676 (C＝O), 1231 (P＝O), 1009 (P—O—C), 1661, 1600, 1537, 1491 (芳环骨架振动) cm－1; MS m/z (%): 211.0 (40.38), 377.0 (M＋H＋, 100), 752.8 (2M＋H＋, 22.63). Anal. calcd for C19H25N2O4P: C 60.63, H 6.69, N 7.44; found C 60.49, H 6.74, N 7.33. 2  结果与讨论 2.1  合成及反应机理 根据文献报道[9]的方法, 用类 Mannich 反应[12,13]制  468 有 机 化 学 Vol. 25, 2005  备得到 α-氨基苄基膦酸二烷基酯的盐酸盐. {苯基-[(吡啶-4-羰基)-氨基]-甲基}-膦酸二烷基酯的制备是由三苯基膦和六氯乙烷法在无氧无水的条件下由稍加修改的文献方法进行, 根据文献报道[6,7], 其反应的机理可能按照 Scheme 2所描述的反应机理进行.  Scheme 2 此反应的主要特点有: 反应条件温和、副产物少、产品易提纯、原料便宜易得、产率高等诸多优点, 是缩合酰胺键的重要方法之一.  2.2  IR光谱 2a～2c 这三个化合物的红外光谱在 1660～1676  cm－1(s)有强的吸收峰, 是羰基的伸展振动吸收峰(νC＝O); 在 3402～3448 cm－1 (s)有强吸收, 是酰胺的 N—H伸展振动吸收峰(νN—H); 1a～1c, 2a～2c 的化合物在 1596～1604 (s)和1511～1546 cm－1 (s)有强吸收, 是芳环碳骨架伸展振动吸收峰(ν芳环); 在 1225～1246 cm－1 (s)有强吸收, 是磷氧双键伸展振动吸收峰(νP＝O); 在 1009～1074 cm－1 (s)有强的双峰吸收峰, 是 P—O—C的伸展振动吸收峰(νP—O—C). 这就表明这些化合物里既有芳环, 羰基, 又有酰胺键、磷氧双键、磷氧键.  2.3  1H NMR谱 化合物 1a～1c的 1H NMR在 δ 5.0附近都显示有双重裂峰, 由于 NH2上的氢被氘代掉, 无法对苄甲基上的氢进行偶合裂分, 因此苄甲基上的氢只被磷裂分成二重, 其偶合常数 JP-CH 为 18 Hz. 化合物 2a～2c 的 1H NMR 在 δ 5.5 附近有一四重裂峰, 从 1H-1H 相关谱和1H-31P 相关谱可以知道: 苄甲基上的氢先被酰胺上的氢裂分, 其偶合常数 JNH-CH是 9 Hz, 然后再被磷裂分, 其偶合常数 JP-CH是 21 Hz, 故在氢谱上看到四重裂峰, 倘若用氘代水取代氘代氯仿, 由于酰胺上的活泼氢被氘代掉, 在氢谱里苄甲基只能观测到被磷裂分的双重峰.  2.4  13C NMR谱 化合物 2a～2c的 13C NMR在 δ 168～165显示吸收峰, 可归属于酰胺键 O＝C—N—H 碳原子, 在 δ 128～133 的峰归属于苯环上的碳, 而吡啶环上的碳其化学环境相差较大, 靠近N端的碳在 δ 149.5～151.5有吸收峰, 与羰基相连的碳在 δ 121～123有吸收峰. 此三个化合物它们的苯环和吡啶环的碳化学环境相差很小, 其吸收峰的化学位移几乎没有改变.  2.5  主要 MS数据分析 表 1 中的数据显示: 这些化合物都有分子离子峰 (M＋H＋)和分子自身二聚峰(2M＋H＋), 化合物 2a～2c的二级质谱都有m/z＝211, m/z＝106的碎片峰, 以 2b化合物为例, 裂解方式可能如 Scheme 3进行. 表 1  化合物 1a～1c和 2a～2c的主要质谱数据  Table 1  The main mass spectral data of compounds 1a～1c and 2a～2c Compound m/z (%) 1a 106.1 (10.64) 215.9 (M＋H＋, 21.60) 430.7 (2M＋H＋, 100) 1b 106.1 (44.21) 243.9 (M＋H＋, 95.44) 486.7 (2M＋H＋, 100 ) 1c 106.1 (17.16) 271.9 (M＋H＋, 66.53) 544.5 (2M＋H＋, 100) 2a 210.9 (1 9.48) 321.0 (M＋H＋, 100) 641.0 (2M＋H＋, 1.01) 2b 211.1 (17.77) 348.9 (M＋H＋, 100) 696.7 (2M＋H＋, 10.16) 2c 211.0 (40.38) 377.0 (M＋H＋, 100) 752.8 (2M＋H＋, 22.63)  Scheme 3    No. 4 方华等：{苯基-[(吡啶-4-羰基)-氨基]-甲基}-膦酸二烷基酯的合成与波谱表征 469  2.6  生物活性分析 2.6.1  抗菌活性 抗菌活性用滤纸片法[14]测定溶解于甲醇的化合物对指示菌的拮抗性, 化合物 1a, 1b, 1c, 2a, 2b对金黄色葡萄球菌(Staphylococcus allreus)有抑菌活性(抑菌圈 d＝7 mm),化合物(1c)对枯草芽孢杆菌(Bzcillus subtilis)有强的抑菌活性(抑菌圈 d＝12 mm).  2.6.2  抗肿瘤活性 抗肿瘤活性采用细胞毒 MTT 法测定[15,16]: 测得2a～2c化合物都具有一定的抗KB(人口腔上皮癌细胞)癌细胞的生物活性, 它们的 IC50值分别为 114.1, 68.5, 51.8 µg/mL, 其活性随着 R 取代基团的增大而增强, 为寻找具有更高活性的含磷化合物指明了方向. References 1 Paolo, F.; Salvatore, F. Phosophorus, Sulfur Silicon Relat. Elem. 1996, 110, 189.  2 Baylis, E. K.; Campbell, C. D.; Dingwall, J. G. J. Chem. Soc., Perkin Trans. 1 1984, 2845.  3 Kowalik, J.; Sawka, D. W.; Gowiak, T. J. Chem. Soc., Chem. Commun. 1984, 446.  4 Livingstone, S. E.; Mihkelson, A. E. Inorg. Chem. 1970, 9 (11), 2545.  5 Chen, R.-Y.; Liu, L.-Z. Study on Organic Phosphorus Chemistry, Higher Education Press, Beijing, 2001, p. 3 (in Chinese). (陈茹玉, 刘纶祖, 有机磷化学研究, 高等教育出版社, 北京, 2001. p. 3.) 6 Appel, R.; Halstenbery, J. Chem. Ber. 1977, 110, 2374.  7 Appel, R.; Heinfried, S. Chem. Ber. 1977, 110, 2382.  8 Appel, R.; Roland, M. Chem. Ber. 1977, 110, 2385.  9 Takahashi, H.; Yoshioka, M.; Imai, N.; Onimura, K. Syn-thesis 1994, 763.  10 Yuan, C.-Y.; Wang, G.-H.; Chen, S.-J. Synthesis 1990, 522.  11 Yuan, C.-Y.; Xi, Y.-M. Acta Chim. Sinica 1986, 44(3), 280 (in Chinese).  (袁承业, 漆又毛, 化学学报, 1986, 44(3), 280.) 12 Chen, R.-Y.; Qing, D. Chin. Chem. Lett. 1995, 6(3), 181.  13 House, H. O.; Trost, B. M. J. Org. Chem. 1964, 29, 1339.  14 Li, G.-L. Microbiology 2001, 28(6), 64 (in Chinese).  15 Zhang, Y.-J.; Wang, J.-F.; Huang, Y.-J.; Zheng, Z.-H.; Su, W.-J. J. Xiamen Univ. (Nat. Sci. Ed.) 2002, 41(6), 804 (in Chinese). (张永杰, 王建锋, 黄耀坚, 郑忠辉, 苏文金, 厦门大学学报(自然科学版), 2002, 41(6), 804.) 16 Mosmann, F. J. Immunol. Methods 1983, 65, 55. (Y0406073  ZHAO, X. J.; ZHENG, G. C.)   VI Chinese Journal of Organic Chemistry Vol. 25, 2005  Synthesis and Spectral Characterization of {Phenyl-[(pyridine-4-carbonyl)amino]- methyl}phosphonic Acid Dialkyl Ester  FANG, Hua; FANG, Mei-Juan; LIU, Xiao- Xia; XU, Peng-Xiang; ZHAO, Yu-Fen* Chin. J. Org. Chem. 2005, 25(4), 466   Compounds 1a～1c and 2a～2c were synthesized via Mannich reaction and peptide coupling reaction. Their structures were confirmed by elemental analysis, IR, 1H NMR, 13C NMR and MS spectra, and the antibacterial and antitumor activities of these com-pounds were given. Study on the Synthesis of Silsesquiox-ane WEN, Yong-Xiang; LIU, An-Hua* Chin. J. Org. Chem. 2005, 25(4), 470   Epoxide silicate substances were derived by creatively using the reaction of functional halogen-containing compounds with reactive pentacoordinate silicon complexes. Bycontrolling the hydrolysis of the epoxide silicate substance and tetraethyl orthocarbon-ate, a novel type of epoxy-functionalized silsesquioxane was prepared. And the synthe-sized product was characterized and analyzed by X-ray diffractometry, fourier transform infrared spectroscopy and differential scanning calorimetry. Nonclassical Condensed Thiophene: Progress in the Study of Thieno[3,4-c]- thiophene GENG, Rong-Xia Chin. J. Org. Chem. 2005, 25(4), 475        Thieno[3,4-c]thiophenes have non-classical structures, the 10π-electron system with sulfur in the formal +4 oxidation state is of interest from both the synthetic and theoretical viewpoints. It has been the object of chemical investigations for almost four decades.  

Synthesis and spectral characterization of {phenyl-[(pyridine-4carbonyl)amino]methyl}phosphonic acid dialkyl ester

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Synthesis and spectral characterization of {phenyl-[(pyridine-4carbonyl)amino]methyl}phosphonic acid dialkyl ester

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