膽酸酰氧基膦酸酯衍生物的合成及抗腫瘤活性

郭深深,代本才,陳　瑨,霍萃萌,劉曉莉,3,趙永德*

(1.河南大學(xué) 天然藥物與免疫工程重點(diǎn)實(shí)驗(yàn)室, 河南 開(kāi)封 475004；　2.河南省科學(xué)院化學(xué)研究所,河南 鄭州 450002；

3.河南大學(xué) 化學(xué)化工學(xué)院, 河南 開(kāi)封 475004)

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膽酸酰氧基膦酸酯衍生物的合成及抗腫瘤活性

郭深深1,2,代本才2,陳瑨2,霍萃萌2,劉曉莉2,3,趙永德1,2*

(1.河南大學(xué) 天然藥物與免疫工程重點(diǎn)實(shí)驗(yàn)室, 河南 開(kāi)封 475004；2.河南省科學(xué)院化學(xué)研究所,河南 鄭州 450002；

3.河南大學(xué) 化學(xué)化工學(xué)院, 河南 開(kāi)封 475004)

摘要：以膽酸和亞磷酸酯為原料,在縮合劑二環(huán)己基碳二亞胺(DCC)和4-二甲基吡啶(DMAP)的催化下進(jìn)行酯化反應(yīng),合成了10個(gè)未見(jiàn)報(bào)道的膽酸酰氧基膦酸酯衍生物,所有目標(biāo)化合物均經(jīng)過(guò)TG, IR,1H NMR, (31)P NMR和HRMS對(duì)其進(jìn)行了結(jié)構(gòu)確認(rèn). 利用MTT法測(cè)定了目標(biāo)化合物的抗腫瘤活性,結(jié)果顯示：目標(biāo)化合物4H, 4I, 4J對(duì)人肝癌細(xì)胞(HepG2)表現(xiàn)出較好的增殖抑制作用.

關(guān)鍵詞：膽酸；α-羥基膦酸酯；酯化反應(yīng)；抗腫瘤活性

膽酸是在哺乳動(dòng)物肝臟中合成的一種多羥基羧酸雙親性分子，三個(gè)羥基和一個(gè)羧基形成親水的α面，而三個(gè)甲基指向另一面形成憎水的β面[1]. 近年來(lái)，隨著人們對(duì)膽酸的探究更加深入，發(fā)現(xiàn)膽酸具有抗菌[2]、抗病毒[3]、抗腫瘤[4-6]等活性. 由于膽酸的羥基和羧基的活性非常高，很容易進(jìn)行修飾，而由膽酸制備的一些衍生物仍然能夠體現(xiàn)出膽酸的一些特性，所以膽酸在藥物化學(xué)[7]、超分子領(lǐng)域[8]以及生物醫(yī)學(xué)[9-10]等領(lǐng)域有著廣泛的應(yīng)用前景.α-羥基膦酸酯作為含磷化合物，在藥物合成中扮演著重要角色，相關(guān)研究發(fā)現(xiàn)膦酸酯類衍生物表現(xiàn)出抗病毒[11]、殺菌[12]、抗腫瘤[13-14]等生物活性. 為了尋找效果更好的抗腫瘤活性分子，通過(guò)酯化反應(yīng)將α-羥基膦酸酯類化合物引入到膽酸分子結(jié)構(gòu)中，設(shè)計(jì)合成了10個(gè)膽酸酰氧基膦酸酯衍生物，這些化合物均未見(jiàn)報(bào)道. 通過(guò)TG, IR,1H NMR,31P NMR和HRMS對(duì)其結(jié)構(gòu)進(jìn)行了表征，并對(duì)該類化合物進(jìn)行了人白血病細(xì)胞(K-562)和人肝癌細(xì)胞(HepG2)的抗腫瘤活性研究. 化合物合成路線見(jiàn)圖1.

圖1　目標(biāo)化合物4A-4J的合成路線Fig.1　Synthetic routes for target compounds 4A-4J

1實(shí)驗(yàn)部分

1.1儀器和試劑

FTS-60型紅外光譜儀(美國(guó)Bio-Rad公司)，KBr壓片；Bruker DPX-300型核磁共振儀(德國(guó)Bruker公司)，溶劑為CDCl3，內(nèi)標(biāo)TMS；Bruker Esquire-3000型質(zhì)譜儀(德國(guó)Bruker公司)；STA-449C型熱重聯(lián)用分析儀(德國(guó)NETSZCH公司).

氟化鉀、膽酸、二環(huán)己基碳二亞胺(DCC)、4-二甲氨基吡啶(DMAP)、亞磷酸二乙酯和亞磷酸二異丙酯均采購(gòu)于阿拉丁試劑公司；四氫呋喃用鈉和二苯甲酮干燥處理.

1.2實(shí)驗(yàn)步驟

1.2.1中間體3的合成

將10 mmol的亞磷酸二乙酯和12 mmol的苯甲醛加入到10 mL的圓底燒瓶中，攪拌均勻后加入20 mmol 的氟化鉀做催化劑. 室溫?cái)嚢?0 min，重結(jié)晶得白色固體3A. 同樣方法[15]合成中間體3B-3J.

O,O′-二乙基-α-苯基-α-羥基甲基膦酸酯(3A): 白色固體2.39 g,產(chǎn)率98.1%；m.p. 83～84℃；IR (KBr，cm-1): 3 262.3, 2 989.6, 1 492.0, 1 449.7, 1 227.7, 1 049.6, 978.7,701.6, 560.2;1H NMR (CDCl3, 300 MHz)δ：7.43 (d,J=7.8 Hz, 2H, ArH), 7.25～7.35 (m, 3H, ArH), 6.18 (d,J=7.5 Hz, 1H, OH), 4.92 (d,J=13.5 Hz, 1H, CH), 3.33～3.99 (m, 4H, 2OCH2), 1.18 (t,J=2.4 Hz, 6H, 2CH3);31P NMR (40.5 MHz, CDCl3)δ：17.756; HRMS:m/z267.079 0 [M+Na]+(計(jì)算值C11H17O4PNa, 267.076 2).

1.2.2目標(biāo)化合物4的合成

將10 mmol的中間體3和15 mmol的膽酸加入到100 mL的圓底燒瓶中，加入40 mL干燥的四氫呋喃溶解. 冰水浴下15 min內(nèi)滴加催化劑(12 mmol的DCC和3 mmol的DMAP溶于20 mL四氫呋喃中). 45 ℃下回流，TLC監(jiān)測(cè)反應(yīng)進(jìn)程，16 h結(jié)束反應(yīng). 減壓蒸去溶劑，得白色固體. 常壓柱層析[V(乙酸乙酯)∶V(石油醚)=1∶5]分離，即得產(chǎn)物.

O,O′-二乙基-α-苯基-α-(3,7,12-三羥基膽甾烷酰氧基)-甲基膦酸酯(4A): 白色固體4.99 g,產(chǎn)率78.8%；熱分解溫度: 254.8 ℃; IR (KBr，cm-1): 3 435.4, 2 932.5, 1 747.9, 1 637.8, 1 472.8, 1 249.1, 1 026.6;1H NMR (CDCl3, 300 MHz)δ： 7.48 (d,J=7.8 Hz, 2H, ArH), 7.34～7.36 (m, 3H, ArH), 6.15 (d,J=13.5 Hz, 1H, CH), 4.05～4.10 (m, 4H, 2OCH2), 3.96 (s, 1H, OCH), 3.84 (s, 1H, OCH), 3.40～3.55 (m, 1H, OCH), 1.25～2.25 (m, 27H, steroidal H), 1.24 (t,J=2.1 Hz, 6H, 2CH3), 0.97 (s, 3H, CH3), 0.90 (s, 3H, CH3), 0.65 (s, 3H, CH3);31P NMR (40.5 MHz, CDCl3)δ：18.158; HRMS:m/z657.351 4 [M+Na]+(計(jì)算值C35H55O8PNa, 657.353 2).

O,O′-二異丙基-α-苯基-α-(3,7,12-三羥基膽甾烷酰氧基)-甲基膦酸酯(4B):白色固體5.05 g,產(chǎn)率76.3%；熱分解溫度: 227.8 ℃; IR (KBr，cm-1): 3 434.0, 2 933.2, 1 748.1, 1 638.1, 1 453.4, 1 248.2, 999.5;1H NMR (CDCl3, 300 MHz)δ：7.48 (d,J=6.6 Hz, 2H, ArH), 7.34～7.36 (m, 3H, ArH), 6.08 (d,J=13.8 Hz, 1H, CH), 4.58～4.66 (m, 2H, 2OCH), 3.95 (s, 1H, OCH), 3.84 (s, 1H, OCH), 3.44～3.65 (m, 1H, OCH), 1.34～2.38 (m, 27H, steroidal H), 1.28 (d,J=6.3 Hz, 6H, 2CH3), 1.23 (d,J=6.3 Hz, 3H, CH3), 1.09 (d,J=6 Hz, 3H, CH3), 0.97 (s, 3H, CH3), 0.89 (s, 3H, CH3), 0.64 (s, 3H, CH3);31P NMR (40.5 MHz,CDCl3)δ：16.248; HRMS:m/z685.381 5 [M+Na]+(計(jì)算值C37H59O8PNa, 685.384 5).

O,O′-二乙基-α-(4-甲基苯基)-α-(3,7,12-三羥基膽甾烷酰氧基)-甲基膦酸酯(4C)：白色固體4.57 g,產(chǎn)率70.6%；熱分解溫度: 238.6 ℃; IR (KBr，cm-1): 3 433.7, 2 933.1, 1 747.8, 1 637.7, 1 446.6, 1 249.8, 1 023.9;1H NMR (CDCl3, 300 MHz)δ：7.38 (q,J1=8.1 Hz,J2=1.8 Hz, 2H, ArH), 7.17 (d,J=8.1 Hz, 2H, ArH), 6.11 (d,J=12.9 Hz, 1H, CH), 4.04～4.10 (m, 4H, 2OCH2), 3.96 (s, 1H, OCH), 3.84 (s, 1H, OCH), 3.40～3.55 (m, 1H, OCH), 2.34 (s, 3H, CH3), 1.30～2.25 (m, 27H, steroidal H), 1.24 (t,J=2.1 Hz, 6H, 2CH3), 0.97 (s, 3H, CH3), 0.89 (s, 3H, CH3), 0.65 (s, 3H, CH3);31P NMR (40.5 MHz, CDCl3)δ：18.319; HRMS:m/z671.368 0 [M+Na]+(計(jì)算值C35H57O8PNa, 671.368 0).

O,O′-二異丙基-α-(4-甲基苯基)-α-(3,7,12-三羥基膽甾烷酰氧基)-甲基膦酸酯(4D)：白色固體5.10 g,產(chǎn)率75.4%；熱分解溫度: 240.8 ℃; IR (KBr，cm-1): 3 434.3, 2 931.9, 1 747.5, 1 637.8, 1 451.3, 1 375.9, 1 249. 0, 1 144.4, 998.7;1H NMR(CDCl3, 300 MHz)δ： 7.37 (q,J1=8.1 Hz,J2=1.8 Hz, 2H, ArH), 7.15 (d,J=8.1 Hz, 2H, ArH), 6.05 (d,J=14.1 Hz, 1H, CH), 4.51～4.60 (m, 2H, OCH), 3.97 (s, 1H, OCH), 3.86 (s, 1H, OCH), 3.83～3.85 (m, 1H, OCH), 2.34 (s, 3H, CH3), 1.34～1.94 (m, 27H, steroidal H), 1.28 (d,J=6.3 Hz, 6H, 2CH3), 1.24 (d,J=6.3 Hz, 3H, CH3), 1.10 (d,J=6.3 Hz, 3H, CH3), 0.97 (s, 3H, CH3), 0.89 (s, 3H, CH3), 0.65 (s, 3H, CH3);31P NMR (40.5 MHz, CDCl3)δ： 16.438；HRMS:m/z699.399 6 [M+Na]+(計(jì)算值C38H61O8PNa, 699.400 2).

O,O′-二乙基-α-(2-氟苯基)-α-(3,7,12-三羥基膽甾烷酰氧基)-甲基膦酸酯(4E):白色固體4.10 g,產(chǎn)率62.9%；熱分解溫度: 235.1 ℃; IR (KBr，cm-1): 3 431.1, 2 935.1, 2 867.7, 1 751.5, 1 617.1, 1 492.7, 1 236.2, 1 028.2;1H NMR (CDCl3, 300 MHz)δ： 7.52～7.55 (m, 1H, ArH), 7.21～7.22 (m, 1H, ArH), 7.19～7.20 (m, 1H, ArH), 7.06～7.07 (m, 1H, ArH), 6.47 (d,J=13.8 Hz, 1H, CH), 3.97～4.18 (m, 4H, 2OCH2), 3.92 (s, 1H, OCH), 3.87 (s, 1H, OCH), 3.40～3.55 (m, 1H, OCH), 1.25～2.21 (m, 27H, steroidal H), 1.23 (t,J=2.1 Hz, 6H, 2CH3), 0.97 (s, 3H, CH3), 0.90 (s, 3H, CH3), 0.66 (s, 3H, CH3);31P NMR (40.5 MHz, CDCl3)δ: 16.438; HRMS:m/z675.343 7 [M+Na]+(計(jì)算值C35H54FO8PNa, 675.343 8).

O,O′-二異丙基-α-(2-氟苯基)-α-(3,7,12-三羥基膽甾烷酰氧基)-甲基膦酸酯(4F)：白色固體3.95 g,產(chǎn)率58.1%；熱分解溫度: 232.3 ℃; IR (KBr，cm-1): 3 430.3, 2 934.4, 2 868.3, 1 751.7, 1 617.4, 1 492.9, 1 384.9, 1 236.6;1H NMR (CDCl3, 300 MHz)δ： 7.56～7.58 (m, 1H, ArH), 7.26～7.28 (m, 1H, ArH), 7.16 (t,J=7.5Hz, 1H, ArH), 7.06 (t,J=9.6Hz, 1H, ArH), 6.39 (d,J=14.1Hz, 1H, CH), 4.74～4.75 (m, 2H, 2OCH), 4.60～4.61 (m, 1H, OCH), 3.94 (s, 1H, OCH), 3.83 (s, 1H, OCH), 1.33～2.46 (m, 27H, steroidal H), 1.27 (m, 6H, 2CH3), 1.24 (d,J=6.3 Hz, 3H, CH3), 1.10 (d,J=6.3 Hz, 3H, CH3), 0.97 (s, 3H, CH3), 0.88 (s, 3H, CH3), 0.64 (s, 3H, CH3);31P NMR (40.5 MHz, CDCl3)δ： 15.581；HRMS:m/z703.373 4 [M+Na]+(計(jì)算值C37H58FO8PNa, 703.375 1).

O,O′-二乙基-α-(3-氟苯基)-α-(3,7,12-三羥基膽甾烷酰氧基)-甲基膦酸酯(4G)：白色固體4.45 g,產(chǎn)率68.2%；熱分解溫度: 240.3 ℃;IR (KBr，cm-1): 3 433.5, 2 934.7, 2 867.6, 1 750.9, 1 592.7, 1 448.7, 1 248.3, 1 142.8, 1 026.0;1H NMR (CDCl3, 300 MHz)δ： 7.33～7.35 (m, 1H, ArH), 7.21～7.27 (m, 2H, ArH), 7.03 (t,J=8.1 Hz, 1H, ArH), 6.13 (d,J=13.8 Hz, 1H, CH), 4.02～4.11 (m, 4H, 2OCH2), 3.96 (s, 1H, OCH), 3.84 (s, 1H, OCH), 3.40～3.55 (m, 1H, OCH), 1.25～2.25 (m, 27H, steroidal H), 1.25 (t,J=2.1 Hz, 6H, 2CH3), 0.97 (s, 3H, CH3), 0.89 (s, 3H, CH3), 0.65 (s, 3H, CH3);31P NMR (40.5 MHz, CDCl3)δ：17.413; HRMS:m/z675.343 3 [M+Na]+(計(jì)算值C35H54FO8PNa, 675.343 8).

O,O′-二異丙基-α-(3-氟苯基)-α-(3,7,12-三羥基膽甾烷酰氧基)-甲基膦酸酯(4H)：白色固體5.03 g,產(chǎn)率73.9%；熱分解溫度: 238.8 ℃; IR (KBr，cm-1): 3 437.8, 2 865.3，1 751.3, 1 592.6, 1 450.1, 1 382.6, 1 245.9, 1 142.6, 999.4;1H NMR (CDCl3, 300 MHz)δ： 7.21～7.33 (m, 3H, ArH), 7.09 (t,J=7.5 Hz, 1H, ArH), 6.06 (d,J=14.1 Hz, 1H, CH), 4.63～4.66 (m, 2H, 2OCH), 3.85～3.96 (m, 2H, 2OCH), 3.44～3.46 (m, 1H, OCH), 1.34～2.38 (m, 27H, steroidal H), 1.29 (d,J=6.3 Hz, 6H, 2CH3), 1.23 (d,J=6.3 Hz, 3H, CH3), 1.14 (d,J=6.3 Hz, 3H, CH3), 0.97 (s, 3H, CH3), 0.89 (s, 3H, CH3), 0.65 (s, 3H, CH3);31P NMR (40.5 MHz, CDCl3)δ： 15.237; HRMS:m/z703.374 7 [M+Na]+(計(jì)算值C37H58FO8PNa, 703.375 1).

O,O′-二乙基-α-(4-氟苯基)-α-(3,7,12-三羥基膽甾烷酰氧基)-甲基膦酸酯(4I)：白色固體4.59 g,產(chǎn)率70.4%；熱分解溫度: 229.8 ℃; IR (KBr，cm-1): 3 427.0, 2 934.2, 2 867.8, 1 749.3, 1 606.4, 1 510.3, 1 376.1, 1 248.5, 1 027.0;1H NMR (CDCl3, 300 MHz)δ： 7.47 (t,J=1.8Hz, 2H, 2OCH), 7.05 (t,J=8.7Hz, 2H, ArH), 6.11 (d,J=13.5Hz, 1H, CH), 4.00～4.12 (m, 4H, 2OCH2), 3.93 (s, 1H, OCH), 3.84 (s, 1H, OCH), 3.43～3.49 (m, 1H, OCH), 1.25～2.25 (m, 27H, steroidal H), 1.28 (t,J=2.1 Hz, 6H, 2CH3), 0.98 (s, 3H, CH3), 0.89 (s, 3H, CH3), 0.65 (s, 3H, CH3);31P NMR (40.5 MHz, CDCl3)δ： 17.546；HRMS:m/z675.340 7 [M+Na]+(計(jì)算值C35H54FO8PNa, 675.343 8).

O,O′-二異丙基-α-(4-氟苯基)-α-(3,7,12-三羥基膽甾烷酰氧基)-甲基膦酸酯(4J)：白色固體5.13g, 產(chǎn)率75.5%；熱分解溫度: 240.6 ℃; IR (KBr，cm-1): 3 436.0, 2 934.6, 2 868.1, 1 749.5, 1 606.5, 1 510.3, 1 466.1, 1 376.1, 1 230.3, 999.2;1H NMR (CDCl3, 300 MHz)δ： 7.47 (q,J1=6.6Hz,J2=1.5Hz, 2H, ArH), 7.04 (t,J=9.3Hz, 2H, ArH), 6.04 (d,J=14.1Hz, 1H, CH), 4.60～4.70 (m, 2H, 2OCH), 3.95 (s, 1H, OCH), 3.85 (s, 1H, OCH), 3.43～3.47 (m, 1H, OCH), 1.34～2.38 (m, 27H, steroidal H), 1.29 (m, 6H, 2CH3), 1.24 (d,J=6.3 Hz, 3H, CH3), 1.09 (d,J=6.3 Hz, 3H, CH3), 0.98 (s, 3H, CH3), 0.90 (s, 3H, CH3), 0.65 (s, 3H, CH3);31P NMR (40.5 MHz, CDCl3)δ： 16.004; HRMS:m/z703.374 7 [M+Na]+(計(jì)算值C37H58FO8PNa, 703.375 1).

1.2.3目標(biāo)化合物的抗腫瘤活性測(cè)試

通過(guò)MTT法測(cè)試目標(biāo)化合物對(duì)人白血病細(xì)胞(K-562)和人肝癌細(xì)胞(HepG2)的抗腫瘤活性，對(duì)照實(shí)驗(yàn)為安菲奈特.

將對(duì)數(shù)生長(zhǎng)期的細(xì)胞制成單細(xì)胞懸液，調(diào)整細(xì)胞個(gè)數(shù)為5×103個(gè)/mL，加到96孔培養(yǎng)板中使其貼壁培養(yǎng)過(guò)夜，24 h后加入不同濃度的樣品液，48 h后加入MTT溶液，每孔100 μL；在37 ℃條件下于培養(yǎng)箱中培養(yǎng)4 h，再除去細(xì)胞上清液，將每孔細(xì)胞結(jié)晶用150 μL DMSO溶解. 分別將樣品組、對(duì)照組(不加樣品)和空白組(只有培養(yǎng)基,無(wú)細(xì)胞)，在酶標(biāo)儀上測(cè)量波長(zhǎng)570 nm處得的光密度(OD)值. 測(cè)得光密度值后由下面的公式計(jì)算出不同樣品濃度下的腫瘤細(xì)胞抑制率.

腫瘤抑制率=(對(duì)照組OD值-樣品組OD值)/

2結(jié)果與討論

2.1目標(biāo)化合物的合成

由于α-羥基膦酸酯是一種活性較弱的親核試劑，空間位阻較大，使得酯化反應(yīng)不易進(jìn)行. 因此，采用不同的催化體系和溶劑來(lái)探索最佳反應(yīng)條件. 以4A為例：分別采用三種催化體系(DMAP、DCC、DCC/DMAP)來(lái)催化反應(yīng). 結(jié)果表明：以DCC/DMAP做催化劑時(shí)產(chǎn)率高達(dá)78.8%(見(jiàn)表1). 同時(shí)又探索了不同溶劑對(duì)該反應(yīng)的影響：分別采用環(huán)己烷、氯仿、四氫呋喃和甲苯四種不同溶劑進(jìn)行反應(yīng)，發(fā)現(xiàn)四氫呋喃和氯仿做溶劑時(shí)產(chǎn)率較高(見(jiàn)表2). 從表2中可以看出，采用中等極性溶劑，反應(yīng)產(chǎn)率要大于較大或較小極性的溶劑.

表1　不同催化體系對(duì)4A產(chǎn)率的影響

表2　不同溶劑對(duì)4A產(chǎn)率的影響

2.2譜圖解析

在IR譜中，酯基中C=O伸縮振動(dòng)吸收接近1 735 cm-1，苯環(huán)在1 450～1 600 cm-1之間有中等強(qiáng)度的骨架振動(dòng)吸收峰，P=O在1 220～1 600 cm-1之間有有較強(qiáng)的伸縮振動(dòng)吸收峰. 通過(guò)1H NMR發(fā)現(xiàn)，在7.0～7.6之間有四個(gè)或五個(gè)氫，符合取代苯的質(zhì)子吸收規(guī)律；另外，與磷原子相連的次甲基上的H由于受到磷原子的影響，在6.0～6.5之間出現(xiàn)一個(gè)標(biāo)準(zhǔn)的雙峰；而磷譜中P的化學(xué)位移出現(xiàn)在16.0～18.5之間，這與文獻(xiàn)報(bào)道一致. HRMS進(jìn)一步證明了目標(biāo)化合物的結(jié)構(gòu).

2.3目標(biāo)化合物的抗腫瘤活性

從表3可以看出，目標(biāo)化合物對(duì)人白血病細(xì)胞(K-562)表現(xiàn)出較弱的增殖抑制活性，但是生長(zhǎng)抑制率并不隨著目標(biāo)物濃度的增高而明顯的增高；且不難發(fā)現(xiàn)，伴隨著目標(biāo)化合物官能團(tuán)的改變，癌細(xì)胞生長(zhǎng)抑制率沒(méi)有明顯的變化.

表4顯示，目標(biāo)化合物對(duì)人肝癌細(xì)胞(HepG2)表現(xiàn)出了一定的增殖抑制活性. 首先， 隨著目標(biāo)化合物濃度的增大，對(duì)HepG2細(xì)胞的生長(zhǎng)抑制率也相應(yīng)的增高；其次，隨著取代基R1的變化，不同目標(biāo)化合物表現(xiàn)出不同的抑制活性. 顯然，含F(xiàn) 的化合物如4E、4F、4G、4H、4I和4J顯示出了更好的抗腫瘤活性. 它對(duì)HepG2細(xì)胞的生長(zhǎng)抑制率已經(jīng)接近甚至超過(guò)對(duì)照藥安菲奈特的效果. 通過(guò)4A與4B，4C與4D，4E與4F，4G與4H，4I與4J的細(xì)胞生長(zhǎng)抑制率的對(duì)比，官能團(tuán)R2為異丙基的目標(biāo)化合物抗腫瘤活性要高于乙基.

總的來(lái)說(shuō)，從表3和表4對(duì)比可以得出，目標(biāo)化合物對(duì)K-562細(xì)胞沒(méi)有明顯的腫瘤細(xì)胞抑制活性. 但是對(duì)HepG2細(xì)胞則體現(xiàn)出一定的增殖抑制活性.

表3　目標(biāo)化合物對(duì)K-562細(xì)胞的生長(zhǎng)抑制率

表4　目標(biāo)化合物對(duì)HepG2細(xì)胞的生長(zhǎng)抑制率

3結(jié)論

以膽酸和中間體α-羥基膦酸酯類化合物為原料，采用DCC/DMAP的催化體系進(jìn)行酯化反應(yīng)，合成了10個(gè)結(jié)構(gòu)新穎的膽酸酰氧基膦酸酯類衍生物. 考察了催化劑，溶劑等條件對(duì)反應(yīng)的影響，得到合適簡(jiǎn)便的反應(yīng)路線,并且研究了目標(biāo)化合物對(duì)腫瘤細(xì)胞的抑制活性,結(jié)果 4H、4I、4J顯示出了較好的抗腫瘤活性.

致謝:感謝新鄉(xiāng)醫(yī)學(xué)院檢驗(yàn)學(xué)院王明永老師及其團(tuán)隊(duì)在目標(biāo)化合物抗腫瘤活性檢測(cè)方面給予的幫助.

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[責(zé)任編輯：任鐵鋼]

Synthesis and antitumor activity of cholic acid-phosphonate derivatives

GUO Shenshen1,2, DAI Bencai2, CHEN Jin2, HUO Cuimeng2, LIU Xiaoli2,3, ZHAO Yongde1,2*

(1.KeyLaboratoryofNaturalMedicineandImmuneEngineering,HenanUniversity,Kaifeng475004,Henan,China;2.InstituteofChemistry,HenanAcademyofSciences,Zhengzhou450002,Henan,China;3.CollegeofChemistryandChemicalEngineering,HenanUniversity,Kaifeng475004,Henan,China)

Abstract:10 cholic acid-phosphonate derivatives based on cholic acid and α-hydroxyphosphonate have been synthesized by using DCC/DMAP condition. These compounds were reported for the first time and confirmed by TG, IR,1H NMR, (31)P NMR and HRMS. And the antitumor activity of the cholic acid derivatives have been measured by MTT assay method. The results showed that the target compounds exhibited certain antitumor activity against HepG2, especially 4H, 4I, 4J.

Keywords:cholic acid; α-hydroxyphosphonate; esterification reaction; antitumor activity

文章編號(hào)：1008-1011(2016)02-0183-06

中圖分類號(hào)：O629.2

文獻(xiàn)標(biāo)志碼：A

作者簡(jiǎn)介：郭深深(1990-)，男，碩士生，研究方向?yàn)樗幬锖铣? *通訊聯(lián)系人, E-mail：963339210@qq.com.

收稿日期：2015-10-25.