《新型小分子类肽氨肽酶N抑制剂的设计、合成与初步活性评价 精灵论文.doc》由会员分享，可在线阅读，更多相关《新型小分子类肽氨肽酶N抑制剂的设计、合成与初步活性评价 精灵论文.doc（9页珍藏版）》请在三一文库上搜索。
1、新型小分子类肽氨肽酶 N 抑制剂的设计、合 成与初步活性评价李荀（山东大学药学院，济南 250012）摘要：目的设计合成 L-异谷氨酰胺类小分子类肽化合物，体外筛选其氨肽酶 N (APN)及 MMP-2 的抑制活性，研究构-效关系。方法通过“拼接原理”，经缩合等反应合成小分子类肽 衍生物，采用体外抑酶活性实验测定目标化合物抑制 APN 与 MMP-2 的活性。结果合成了22 个未见文献报道的化合物，结构经红外光谱、核磁共振氢谱、质谱确证。结论这类化合 物的氨肽酶 N 抑制活性显著高于 MMP-2，为选择性较高的氨肽酶 N 抑制剂。其中，化合物8 (IC50 = 10.2 0.9 M)的活性显著
2、高于阳性对照药 Bestatin (IC50 = 13.1 0.7 M),有望 成为先导物做进一步的研究。关键词： 药物化学；APN/CD13; 抑制剂; Peptidomimetic; AnticancerDesign, Synthesis and Preliminary Activity Evaluation ofNovel Peptidomimetics as Aminopeptidase N/CD13InhibitorsLi Xun(College of Pharmacy, Shandong University, Jinan250012)Abstract: Aim To design
3、 and synthesize novel peptidomimetic analogues evaluated their inhibitory activities against aminopeptidase N (APN/CD13) and matrix metalloproteinase-2 (MMP-2). Methods According to the “combination principles”, peptidomimetic analogues were synthesized through condensation reaction, and their enzym
4、atic inhibitory activities were assayed in vitro. Results 22 peptidomimetic APN inhibitors were prepared which have not been reported in literatures. Their structures were confirmed by IR, 1H-NMR, and MS. Conclusion Most of the compounds displayedselective inhibition against APN as compared with MMP
5、-2, with IC50 values in micromole range. Within this series, compound 8 (IC50 = 10.2 0.9 M) demonstrated comparable APN inhibitory activities as compared with the positive control bestatin (IC50 = 13.1 0.7 M), which may providea promising lead for further molecular optimizations.Key words: Medicinal
6、 Chemistry; APN/CD13; inhibitor; Peptidomimetic; Anticancer0引言氨肽酶 N (APN, EC 3.4.11.2)是一种含锌离子的膜结合型外肽酶, 已经证明其在肿瘤细 胞表面大量表达，对肿瘤的侵袭和血管生成有重要的作用1,2。此外，它还能够表达于抗原 递呈细胞表面，降解多种免疫活性物质，使机体免疫力下降，削弱巨噬细胞和 NK 细胞对肿 瘤细胞的识别和杀伤能力3,4。该酶还可降解细胞外基质（ECM）的主要成分，促进肿瘤细 胞的生长和转移5,6。因此，抑制该酶的活性就可对肿瘤的侵袭和转移以及血管生成进行有 效地控制，同时增强粒细胞的趋化性，
7、提高机体的免疫能力，进而可能成为有效的抗癌或抗 炎药物7-9。截至目前，已经有多种 APN 抑制剂类药物进入临床10，其中，Bestatin 作为抗 白血病的治疗药物已经在日本上市多年11，本实验将采用其作为阳性对照12。基金项目：博士点青年基金（20070422061）；山东省自然基金（Y2008C01）；山东大学自主创新基金(2009TS113)作者简介：李荀（1976.7），女，副教授，抗肿瘤药物的设计合成. E-mail: 传统的以酶为靶点的肽类化合物有很多缺点，例如对蛋白水解酶不稳定、生物利用度低、 作用时间短而排泄快等。因而目前大多数报道的 APN 抑制剂采用了模拟肽（peptid
8、emimics） 的结构。它是一种利用人工合成的物质来模拟天然多肽的结构，或者利用构象型模板诱导相 邻的多肽序列而形成的具特异性结构的化合物。因此，设计合成模拟肽类化合物是发现酶抑 制剂类药物的理想策略13,14。本文根据 APN 三维结构特征，结合计算机辅助药物设计，通过药物设计中的“拼接原 理”，将具有抗肿瘤活性的没食子酸和天然 L-异谷氨酸胺利用缩合反应进行对接定向合成， 再引入各种天然氨基酸片段，合成了 22 个未见文献报道的新型小分子类肽化合物，合成路 线见图 1。该法以 DCC 为缩合剂，反应条件温和、操作简单。1合成熔点用 X-4 型数字显微熔点仪测定，温度未校正。IR 谱采用
9、FTIT-8400 型红外分光光 度计测定，KBr 压片。1H-NMR 谱采用 Bruker400 型核磁共振谱仪测定，DMSO-d6 为溶剂， TMS 为内标。生物活性测定用试剂如 APN，MMP-2，L-亮氨酰对硝基苯胺均购自 Sigma 公 司，使用前超低温冷藏保存备用。实验中其余试剂均为市售分析纯或化学纯，薄层色谱用硅 胶 GF254 由青岛海洋生产。OHOOHH3COOOHH3COOClH3COOCOOHNCOOHabcHHOH3COOHOCH3H3COOCH3H3COOCH31234COOH3 O OH3COOdN1HOH3COH3COeH3COOR1NR2NHOCOOCH3OCH
10、3various nucleophilic agentsOCH356a-r, 7HO N COOH OH H H OH3CO H3CON HOCH3N R2 fO COOCH3H3CO H3CON HOCH3N OH NH O R26a R2=H6b R2=CH38 R2=H9 R2=CH3图 1 目标化合物的反应路线示意图Fig. 1 Reaction route of target compounds: a) Me2SO4, 40 % NaOH, then 1 N HCl. b) SOCl2 in benzene. c)L-Glu-Na, Na2CO3, then 2N HCl. d) A
11、c2O, 5560 C. e) L-amino acid methyl ester hydrochloride, anhydrousCH2Cl2, Et3N, rt. f) NH2OK in anhydrous CH3OH, then acetic acid (two steps).1.1 3,4,5-三甲氧基苯甲酸(2)的制备室温下，将没食子酸(1, 5g, 29.4 mmol)溶于 50 mL 4 N NaOH 溶液中，滴加 (CH3)2SO4 (8.9 g, 71 mmol)，在 30-40 C 继续反应 20 分钟, 继续加入没食子酸(5 克) 的 50 mL 4 N NaOH 溶液。
12、将此溶液缓慢升温至 90 C 反应 1 h 后回流 2h，冷却至室温，用 2 N HCl 酸化至 pH 2, 过滤,用蒸馏水洗涤沉淀。粗品再用 50% EtOH 重结晶的白色针状晶体的目标化合 物 2 (40.8 g, 65%)：熔点 168-171 C. 1H NMR 3.97 (s, 9H), 7.23 (s, 2H), 12.11 (s, 1H).1.23,4,5-三甲氧基苯甲酰氯(3)将上述化合物 2 (21.2 g, 100 mmol)溶于 320 mL 苯中，逐滴滴加 SOCl2 (40 mL, 548 mmol)，回流反应 3 h, 减压蒸除溶剂得到黄色油状物 3，继续加入 50
13、 mL 苯后减压整除过量 的 SOCl2，所得的粗品不经纯化直接用于下一步反应中。1.3(R)-2-(3,4,5-三甲氧基苯甲酰氨基)戊二酸(4)-5 C 下，将无水 Na2CO3 (19.6 g, 185 mmol)、L-谷氨酸(16.9 g, 115 mmol) 溶于 150 mL 蒸馏水中，逐滴加入 化合物 3 的 200 mL 苯溶液， 1 h 滴加完毕。反应约 5 小时后，用 2 N HCl 酸化至 pH 5-6, 水相用 2 N HCl 酸化至 pH 3-4, 在冰箱中将有机相（氯仿）放置过夜， 将析出的白色固体用蒸馏水洗涤三次，干燥后的目标化合物 4 (28.6 g, 89.3%
14、)。熔点 120-121C, IR (KBr, cm-1): 3299.7 & 3255.4 (NH), 2942.5 (CH), 1744.7 & 1699.5 (C=O), 1500.7 ( NH),1235.5 & 1127.7 (C-O). 1H NMR (DMSO-d6, ppm): 12.41 (s, 2H, 2-COOH), 8.53 (d, 1H, J =7.2 Hz, NH), 7.22 (s, 2H, Ar-H), 4.41 (m, 1H, CH), 3.83 (s, 6H, 2-OCH3), 3.71 (s, 3H, OCH3),2.35 (m, 2H, CH2), 2.
15、21 (m, 1H, CH), 1.96 (m, 1H, CH). ESI-MS: m/z (rel intensity) 341.8.1.4(R)-N-(2,6-二氧-四氢-2H-吡喃-3-基)-3,4,5-三甲氧基苯甲酰胺(5)在 55-60C 下，将 10 g (2.9 mmol)化合物 4 和 80 mL 重蒸过的醋酸酐置于 250-mL 的圆 底烧瓶中反应 5 h。 然后快速趁热过滤掉体系中的不溶物，然后加入 10 mL 无水乙醚，滤 液冷却至室温，析出白色固体，过滤，干燥后得化合物 5 (5.2 g, 55%). 熔点 150-152 C, IR (KBr, cm-1): 3310
16、.0 (NH), 2945.1 (CH), 1777.0 & 1640.7 (O=C-O-C=O), 1504.2 ( NH), 1239.6& 1129.6 (C-O). 1H NMR (DMSO-d6, ppm): 6.99 (s, 2H, Ar-H), 8.13 (d, 1H, J = 7.2 Hz, NH),4.45 (m, 1H, CH), 3.73 (s, 9H, 3-OCH3), 1.96-2.25 (m, 2H, CH2), 2.18-2.28 (m, 2H, CH2). ESI-MS: m/z (rel intensity) 323.8.1.5L-氨基酸甲酯盐酸盐以甘氨酸甲酯
17、盐酸盐为例：0C 下，甘氨酸(15.0 g, 200 mmol)和 150 mL 无水甲醇的混 悬液通入干燥的 HCl 气体直至混悬液澄清。将此溶液置于冰箱中过夜，蒸出溶剂并加入少 量无水甲醇带走多余的 HCl 气体，将所得的白色固体用 1:4 的无水甲醇/乙醚重结晶得纯品20g, 80%。熔点 175-176 C, IR (KBr, cm-1): 3215.0-3523.0 (m, NH), 2935.2 (CH), 1676.3 (C=O),1129.3 (C-O).1.6(R)-5-(2-甲氧基-2-氧基乙氨基)-5-氧-4-(3,4,5-三甲氧基苯甲酰胺基)戊酸(6a)将化合物 5 (
18、3.23 g, 10 mmol) 和氨基酸甲酯盐酸盐(2.5 g, 20 mmol)溶于 30 mL CH2Cl2 中，加入 mL Et3N，室温下反应直至反应物消失，蒸出溶剂，所得物倒入 50 mL 冰水中， 用 2 N HCl 酸化至 pH 2。置于冰箱中过夜，过滤出所得的白色鳞片状晶体，用冰水洗涤， 干燥，得化合物 6a (0.94 g, 74%). 熔点 169-172 C, IR (KBr, cm-1): 3325.1 (NH), 2939.6 (CH),1745.7 & 1714.5 (O=C-NH), 1637.8 (O=C-O), 1584.3 (C=C), 1131.3 (C
19、-O). 1H NMR (DMSO-d6, ppm): 8.45 (d, 1H, J = 7.5 Hz, NH), 8.38 (t, 1H, J = 5.4 Hz, NH), 7.23 (s, 2H, Ar-H), 4.47 (m,1H, CH), 3.88 (d, 2H, J = 5.4 Hz, CH2), 3.83 (s, 6H, 2-OCH3), 3.69 (s, 3H, OCH3), 3.62 (s, 3H, COOCH3), 2.34 (t, 2H, J = 8.0 Hz, CH2), 2.06, 1.92 (m, 2H, CH2). ESI-MS: m/z (rel intensi
20、ty)412.8.1.7(R)-5-(S)-1-甲氧基-1-氧基丙烷-2-氨基)-5-氧-4-(3,4,5-三甲氧基苯甲酰胺基)戊酸(6b)制备方法同 6a，产率 35.0%, 熔点 153-155 C, IR (KBr, cm-1): 3304.8 (NH), 2933.5 (CH),1751.1 & 1719.9 (O=C-NH), 1647.7 (O=C-O), 1584.2 (C=C), 1129.9 (C-O). 1H NMR (DMSO-d6, ppm): 8.74 (d, 1H, J = 7.8 Hz, NH), 8.33 (d, 1H, J = 6.0 Hz, NH), 7.2
21、4 (s, 2H, Ar-H), 4.36 (m,1H, CH), 4.23 (m, 1H, CH), 3.85 (s, 6H, 2-OCH3), 3.73 (s, 3H, OCH3), 3.60 (s, 3H, COOCH3),2.23 (t, 2H, J = 7.8 Hz, CH2), 2.06, 1.83 (m, 2H, CH2), 1.23 (d, 3H, J = 7.2 Hz, CH3). ESI-MS:m/z (rel intensity) 425.1.1.8(R)-5-(S)-1-甲氧基-1-氧基丙烷-2-氨基)-5-氧-4-(3,4,5-三甲氧基苯甲酰胺基)戊酸(6c)制备方法
22、同 6a，产率 28.5%, 熔点 105-107 C, IR (KBr, cm-1): 3305.2 (NH), 2942.2 (CH),1746.1 & 1717.9 (O=C-NH), 1644.7 (O=C-O), 1586.7 (C=C), 1127.6 (C-O). 1H NMR (DMSO-d6, ppm): 8.73 (d, 1H, J = 7.8 Hz, NH), 8.37 (d, 1H, J = 6.0 Hz, NH), 7.00 (s, 2H, Ar-H), 4.36 (m,1H, CH), 4.27 (m, 1H, CH), 3.73 (s, 6H, 2-OCH3),
23、3.67 (s, 3H, OCH3), 3.82 (s, 3H, COOCH3),2.23 (t, 2H, J = 7.0 Hz, CH2), 2.06, 1.83 (m, 2H, CH2), 1.92 (m, 2H, CH2), 0.98 (d, 3H, J = 7.2 Hz, CH3). ESI-MS: m/z (rel intensity) 439.4.1.9(R)-5-(S)-1-甲氧基-1-氧基丙烷-2-氨基)-5-氧-4-(3,4,5-三甲氧基苯甲酰胺基)戊酸(6d)制备方法同 6a，产率 23.4%, 熔点 110-112 C, IR (KBr, cm-1): 3307.8 (N
24、H), 2940.6 (CH),1755.4 & 1718.1 (O=C-NH), 1640.9 (O=C-O), 1589.2 (C=C), 1125.5 (C-O). 1H NMR (DMSO-d6, ppm): 8.88 (d, 1H, J = 7.8 Hz, NH), 8.24 (d, 1H, J = 8.1 Hz, NH), 6.98 (s, 2H, Ar-H), 4.33 (m,1H, CH), 4.53 (m, 1H, CH), 3.73 (s, 6H, 2-OCH3), 3.62 (s, 3H, OCH3), 3.72 (s, 3H, COOCH3),2.10 (t, 2H,
25、J = 15.3 Hz, CH2), 2.11, 1.79 (m, 2H, CH2), 1.32 (m, 2H, CH2), 1.90 (m, 2H, CH2),0.97 (d, 3H, J = 8.4 Hz, CH3). ESI-MS: m/z (rel intensity) 453.3.1.10(R)-5-(S)-1-甲氧基-1-氧基丙烷-2-氨基)-5-氧-4-(3,4,5-三甲氧基苯甲酰胺 基)戊酸(6e)制备方法同 6a，产率 61.2%, 熔点 99-101 C, IR (KBr, cm-1): 3321.4 & 3256.9 (NH),2956.2 (CH), 1724.1 (O
26、=C-NH), 1647.6 (O=C-O), 1584.3 (C=C), 1130.5 (C-O). 1H NMR (DMSO-d6, ppm): 7.82 (d, 1H, J = 7.6 Hz, NH), 7.15 (s, 1H, Ar-H), 7.12 (s, 1H, Ar-H), 6.44 (d,1H, J = 7.2 Hz, NH), 4.93 (m, 1H, CH), 4.60 (m, 1H, CH), 3.92 (s, 6H, 2-OCH3), 3.80 (s, 3H,OCH3), 3.68 (s, 3H, COOCH3), 2.28 (t, 2H, J = 7.2 Hz, CH
27、2), 2.06 (m, 2H, CH2), 1.92 (m, 2H, CH2), 1.29 (br, 4H, 2-CH2), 0.87 (t, 3H, J = 7.8 Hz, CH3). ESI-MS: m/z (rel intensity) 467.1.1.11 (R)-5-(S)-1-甲氧基-1-氧基-3-苯基丙烷-2-氨基)-5-氧-4-(3,4,5-三甲氧基苯甲 酰胺基)戊酸(6f)制备方法同 6a，产率 56.3%, 熔点 141-144 C, IR (KBr, cm-1): 3416.0 & 3303.7 (NH),2939.9 (CH), 1743.7 & 1718.8 (O=
28、C-NH), 1646.4 (O=C-O), 1584.1 & 1536.7 (C=C), 1128.9 (C-O). 1H NMR (DMSO-d6, ppm): 8.69 (d, 1H, J = 4.3 Hz, NH), 8.38 (d, 1H, J = 6.8 Hz, NH),7.24 (m, 2H, Ar-H), 7.18 (m, 5H, Ar-H), 4.44 (m, 1H, CH), 4.34 (m, 1H, CH), 3.85 (s, 6H,2-OCH3), 3.70 (s, 3H, OCH3), 3.57 (s, 3H, COOCH3), 2.18 (t, 2H, J = 7.
29、6 Hz, CH2), 2.11 (d, 2H, J= 7.4 Hz, CH2), 1.98, 1.76 (2m, 2H, CH2). ESI-MS: m/z (rel intensity) 501.1.1.12(R)-5-(S)-3-(4-氟苯基)-1-甲氧基-1-氧基丙烷-2-氨基)-5-氧-4-(3,4,5-三甲氧 基苯甲酰胺基)戊酸(6g)制备方法同 6a，产率 61.4%, 熔点 107-111 C, IR (KBr, cm-1): 3296.3 (NH), 2942.1 (CH),1743.0 & 1720.4 (O=C-NH), 1648.6 (O=C-O), 1584.2 &
30、1537.3 (C=C), 1127.6 (C-O). 1H NMR (DMSO-d6, ppm): 7.78 (d, 1H, J = 6.6 Hz, NH), 7.26 (s, 2H, Ar-H), 7.12 (m, 2H, Ar-H), 6.95 (m,2H, Ar-H), 6.47 (d, 1H, J = 8.0 Hz, NH), 4.78 (m, 1H, CH), 4.57 (m, 1H, CH), 3.93 (s, 6H,2-OCH3), 3.86 (s, 3H, OCH3), 3.68 (s, 3H, COOCH3), 3.10 (d, 2H, J = 8.2 Hz, CH2),
31、2.27 (t, 2H, J= 7.2 Hz, CH2), 1.89 (m, 2H, CH2). ESI-MS: m/z (rel intensity) 519.2.1.13(R)-5-(S)-3-(4-氯苯基)-1-甲氧基-1-氧基丙烷-2-氨基)-5-氧-4-(3,4,5-三甲氧 基苯甲酰胺基)戊酸(6h)制备方法同 6a，产率 67.2%, 熔点 143-145 C, IR (KBr, cm-1): 3300.1 (NH), 2942.3 (CH),1742.9 & 1718.6 (O=C-NH), 1647.0 (O=C-O), 1584.0 & 1536.9 (C=C), 1129.
32、0 (C-O). 1H NMR (DMSO-d6, ppm): 7.74 (d, 1H, J = 6.4 Hz, NH), 7.29 (s, 2H, Ar-H), 7.22 (m, 2H, Ar-H), 7.10 (m,2H, Ar-H), 6.54 (d, 1H, J = 7.8 Hz, NH), 4.79 (m, 1H, CH), 4.55 (m, 1H, CH), 3.93 (s, 6H,2-OCH3), 3.89 (s, 3H, OCH3), 3.72 (s, 3H, COOCH3), 3.12 (d, 2H, J = 7.6 Hz, CH2), 2.31 (t, 2H, J= 7.6
33、 Hz, CH2), 1.95 (m, 2H, CH2). ESI-MS: m/z (rel intensity) 535.1.1.14(R)-5-(S)-3-(4-溴苯基)-1-甲氧基-1-氧基丙烷-2-氨基)-5-氧-4-(3,4,5-三甲氧 基苯甲酰胺基)戊酸(6i)制备方法同 6a，产率 50.7%, 熔点 166-168 C, IR (KBr, cm-1): 3303.4 (NH), 2946.1 (CH),1744.2 & 1719.0 (O=C-NH), 1647.4 (O=C-O), 1583.8 & 1534.6 (C=C), 1128.7 (C-O). 1H NMR (DM
34、SO-d6, ppm): 7.77 (d, 1H, J = 6.8 Hz, NH), 7.26 (s, 2H, Ar-H), 7.18 (m, 2H, Ar-H), 7.05 (m,2H, Ar-H), 6.62 (d, 1H, J = 4.5 Hz, NH), 4.83 (m, 1H, CH), 4.57 (m, 1H, CH), 3.91 (s, 6H,2-OCH3), 3.80 (s, 3H, OCH3), 3.77 (s, 3H, COOCH3), 3.01 (d, 2H, J = 8.4 Hz, CH2), 2.39 (t, 2H, J= 5.8 Hz, CH2), 1.98 (m,
35、 2H, CH2). ESI-MS: m/z (rel intensity) 580.3.1.15(R)-5-(S)-3-(4-氢苯基)-1-甲氧基-1-氧基丙烷-2-氨基)-5-氧-4-(3,4,5-三甲氧 基苯甲酰胺基)戊酸(6j)制备方法同 6a，产率 49.0%, 熔点 87-89 C, IR (KBr, cm-1): 3355.2 & 3300.7 (NH), 2952.3 (CH), 1744.1 (O=C-NH), 1647.7 (O=C-O), 1585.9 & 1498.7 (C=C), 1257.7 & 1126.5 (C-O). 1H NMR (DMSO-d6, ppm)
36、: 8.72 (d, 1H, J = 7.0 Hz, NH), 8.33 (d, 1H, J = 6.4 Hz, NH), 7.23 (d, 2H, J = 3.4 Hz, Ar-H), 6.95 (m, 2H, Ar-H), 6.64 (m, 2H, Ar-H), 4.34 (m, 1H, CH), 3.83 (s, 6H,2-OCH3), 3.70 (s, 3H, OCH3), 3.56 (s, 3H, COOCH3), 3.49 (m, 1H, CH), 2.20 (t, 2H, J = 7.5 Hz, CH2), 2.11 (d, 2H, J = 6.8 Hz, CH2), 2.00,
37、 1.78 (2m, 2H, CH2). ESI-MS: m/z (rel intensity) 517.1.1.16(R)-5-(S)-3-羟基-1-甲氧基-1-氧基丙烷-2-氨基)-5-氧-4-(3,4,5-三甲氧基苯 甲酰胺基)戊酸(6k)制备方法同 6a，产率 60.0%, 熔点 85-87 C, IR (KBr, cm-1): 3266.1 (NH), 2941.7 (CH),1744.2 (O=C-NH), 1632.2 (O=C-O), 1584.3 (C=C), 1129.3 (C-O). 1H NMR (DMSO-d6, ppm): 8.76 (d, 1H, J = 6.5
38、Hz, NH), 8.26 (d, 1H, J = 6.8 Hz, NH), 7.24 (m, 2H, Ar-H), 5.02 (m, 1H, CH),4.33 (m, 1H, CH), 3.82 (s, 6H, 2-OCH3), 3.71 (s, 3H, OCH3), 3.61 (s, 3H, COOCH3), 3.05 (d, 2H, J = 5.5 Hz, CH2), 2.28 (t, 2H, J = 6.0 Hz, CH2), 2.05, 1.83 (2m, 2H, CH2). ESI-MS: m/z (rel intensity) 441.1.1.17(R)-5-(S)-1-甲氧基-
39、4-(甲基硫基)-1-氧基丁基-2-氨基)-5-氧-4-(3,4,5-三甲氧 基苯甲酰胺基)戊酸(6l)制备方法同 6a，产率 59.1%, 熔点 124-127 C, IR (KBr, cm-1): 3289.3 (NH), 2941.8 (CH),1750.9 & 1717.8 (O=C-NH), 1647.2 (O=C-O), 1584.5 (C=C), 1129.7 (C-O). 1H NMR (DMSO-d6, ppm): 8.72 (d, 1H, J = 5.7 Hz, NH), 8.32 (d, 1H, J = 7.3 Hz, NH), 7.24 (s, 2H, Ar-H), 4
40、.47 (m,1H, CH), 4.37 (m, 1H, CH), 3.78 (s, 6H, 2-OCH3), 3.70 (s, 3H, OCH3), 3.61 (s, 3H, COOCH3),2.26 (t, 2H, J = 7.8 Hz, CH2), 2.13 (t, 2H, J = 7.4 Hz, CH2), 2.06 (m, 4H, 2-CH2), 1.83 (s, 3H, CH3). ESI-MS: m/z (rel intensity) 486.1.1.18(R)-5-(R)-1-甲氧基-1-氧基-3-(苯基硫基)丙烷-2-氨基)-5-氧-4-(3,4,5-三甲氧 基苯甲酰胺基)戊
41、酸(6m)制备方法同 6a，产率 54.0%, 熔点 77-79 C, IR (KBr, cm-1): 3272.9 (NH), 2940.6 (CH),1746.5 & 1722.0 (O=C-NH), 1648.5 (O=C-O), 1584.0 (C=C), 1128.4 (C-O). 1H NMR (DMSO-d6, ppm): 8.77 (d, 1H, J = 6.8 Hz, NH), 8.46 (d, 1H, J = 7.2 Hz, NH), 7.34 (d, 2H, J = 4.1 Hz, Ar-H),7.30 (m, 5H, Ar-H), 4.47 (m, 1H, CH), 4
42、.20 (m, 1H, CH), 3.83 (s, 6H, 2-OCH3), 3.75 (s, 3H, OCH3), 3.60 (s, 3H, COOCH3), 3.32 (s, 2H, CH2), 3.06 (d, 2H, J = 6.8 Hz, CH2), 2.26 (t, 2H, J =7.8 Hz, CH2), 2.07 & 1.84 (2m, 2H, CH2). ESI-MS: m/z (rel intensity) 547.1.1.19(R)-5-(S)-5-(苄氧羰基)-1-甲氧基-1-氧基戊基-2-氨基)-5-氧-4-(3,4,5-三甲氧 基苯甲酰胺基)戊酸(6n)制备方法同
43、6a，产率 43.0%, 熔点 108-112 C, IR (KBr, cm-1): 3333.2 & 3276.9 (NH),2945.9 (CH), 1745.0 & 1718.6 (O=C-NH), 1688.4, 1584.1 (C=C), 1128.2 (C-O). 1H NMR (DMSO-d6, ppm): 8.74 (d, 1H, J = 5.6 Hz, NH), 8.28 (d, 1H, J = 6.9 Hz, NH), 7.34 (m, 5H, Ar-H), 7.24 (s, 2H, Ar-H), 4.99 (s, 2H, CH2), 4.36 (m, 1H, CH), 4
44、.19 (m, 1H, CH), 3.83 (s, 6H,2-OCH3), 3.71 (s, 3H, OCH3), 3.59 (s, 3H, COOCH3), 2.96 (t, 2H, J = 5.2 Hz, CH2), 2.25 (t, 2H, J= 7.8 Hz, CH2), 2.06 & 1.83 (2m, 2H, CH2), 1.66 & 1.54 (2m, 2H, CH2), 1.45 (m, 2H, CH2). ESI-MS: m/z (rel intensity) 602.1.1.20(R)-5-(S)-6-(苄氧羰基)-1-甲氧基-1-氧基己基-2-氨基)-5-氧-4-(3,4
45、,5-三甲氧 基苯甲酰胺基)戊酸(6o)制备方法同 6a，产率 48.0%, 熔点 147-150 C, IR (KBr, cm-1): 3325.6 (NH), 2940.1 (CH),1720.9 (O=C-NH), 1648.8 (O=C-O), 1584.8 & 1498.2 (C=C), 1234.6 & 1126.5 (C-O). 1H NMR (DMSO-d6, ppm): 8.72 (t, 1H, J = 6.0 Hz, NH), 8.24 (t, 1H, J = 7.2 Hz, NH), 7.33 (m, 5H, Ar-H), 7.24 (s, 2H, Ar-H), 7.20
46、 (s, 1H, Ar-H), 4.99 (s, 2H, CH2), 4.37 (m, 1H, CH), 4.17 (m, 1H, CH), 3.83 (s, 6H, 2-OCH3), 3.74 (s, 3H, OCH3), 3.61 (s, 3H, COOCH3), 2.94 (t, 2H, J = 6.0 Hz, CH2), 2.27 (t, 2H, J = 7.8 Hz, CH2), 2.13 (s, 3H, CH3), 2.05 (m, 1H, CH), 1.85 (m, 1H, CH), 1.56 (m, 2H, CH2), 1.36 (m, 2H, CH2), 1.26 (m, 2
47、H, CH2). ESI-MS: m/z (rel intensity) 616.1.1.21(R)-5-(S)-1-甲氧基-5-(3-硝基胍基)-1-氧基戊基-2-氨基)-5-氧-4-(3,4,5-三甲 氧基苯甲酰胺基)戊酸(6p)制备方法同 6a，产率 52.2%, 熔点 123-126 C, IR (KBr, cm-1): 3307.4 (NH), 2943.8 (CH),1739.8 (O=C-NH), 1649.1 (O=C-O), 1584.7 & 1499.7 (C=C), 1236.4 & 1127.5 (C-O). 1H NMR (DMSO-d6, ppm): 8.66 (d, 1H, J = 6.6 Hz, NH), 8.22 (t, 1H, J = 6.0 Hz, NH), 7.24 (s, 2H, Ar-H),4.37 (m, 1H, CH), 4.24 (m, 1H, CH), 3.84 (s, 6H, 2-O