1,4-二取代-1,2,3-三唑衍生物的合成*

于金蘭, 武欽佩, 張青山, 周智明

(北京理工大學(xué) 化工與環(huán)境學(xué)院,北京 100081)

核苷類(lèi)似物因?yàn)榫哂锌共《尽⒖拱┑纳锘钚砸鹆巳藗兊膹V泛關(guān)注，在核苷類(lèi)似物中，其中無(wú)環(huán)糖基的核苷顯示了良好的抗病毒活性，如:Acyclovir[1], Ganciclovir[2]和Cidofovir[3～5]。盡管無(wú)糖環(huán)類(lèi)核苷有良好的抗病毒活性，但這類(lèi)化合物在臨床使用時(shí)存在低的口服生物利用度和難以轉(zhuǎn)移進(jìn)細(xì)胞等缺點(diǎn)[6,7]。因?qū)@類(lèi)化合物進(jìn)行分子修飾以提高其治療指數(shù)已經(jīng)成為新的抗病毒藥物的研究方向。

近年來(lái)，三唑的藥用價(jià)值越來(lái)越受到關(guān)注，經(jīng)常被作為有效官能團(tuán)介入多肽,DNA, RNA和糖類(lèi)化合物中。如Camarasa研究小組[8]利用1,3-偶極環(huán)加成反應(yīng)，合成了抗HIV-1的TSAO-T三唑衍生物。

本文以Cidofovir為原型，在超聲波輻射下,端基炔與疊氮基[3,3-二甲基-2，4-二氧戊環(huán)基]甲烷(4)通過(guò)1,3-偶極環(huán)加成合成了1-(3,3-二甲基-2，4-二氧戊環(huán)基甲基)-4-芳基-1,2,3-三唑(5a～5e); 5在酸性條件下脫保護(hù)得1-(2,3-二羥基丙基)-4-芳基-1，2，3-三唑(6a～6e),其結(jié)構(gòu)經(jīng)1H NMR,13C NMR， MS和元素分析表征。

CompabcdeAr-F-OMe---CH2N2'ONH6'5'4'O

Scheme1

1 實(shí)驗(yàn)部分

1．1 儀器與試劑

XT4A型熔點(diǎn)儀(溫度計(jì)未校正);Bruker Avance型核磁共振儀(DMSO-d6為溶劑,TMS為內(nèi)標(biāo));Bruker Apex Ⅳ FTMS型質(zhì)譜儀；Yanaco MT 23型CHN元素自動(dòng)分析儀；KQ-3200DB型超聲波清洗器。

端基炔,Alfa Aesar公司；其余所用試劑均為分析純。

1.2 合成

(1) 3,3-二甲基-2，4-二氧雜環(huán)戊甲醇(2)的合成

在反應(yīng)瓶中加入3,3-二甲基-2，4-二氧雜環(huán)戊甲醛(1) 6.00 g(46.15 mmol)的甲醇(15 mL)溶液,冰水浴冷卻,攪拌下于0 ℃緩慢滴加NaBH43.00 g(81.08 mmol)的甲醇(15 mL)溶液，反應(yīng)30 min(TLC監(jiān)測(cè))。加水8 mL，蒸去甲醇，殘余物用乙酸乙酯(3×5 mL)萃取，合并有機(jī)相，用無(wú)水硫酸鈉干燥，旋干溶劑得無(wú)色油狀液體2 5.88 g,收率98%。

(2) 1-對(duì)甲苯磺酰氧甲基-3，3-二甲基-2，4-二氧雜環(huán)戊烷(3)的合成

在反應(yīng)瓶中加入24.97 g(37.65 mmol)和三乙胺5.60 mL(40.00 mmol) 的二氯甲烷(120 mL)溶液,對(duì)甲苯磺酰氯1.4 mL(15.00 mmol),攪拌下于室溫反應(yīng)5 h(TLC監(jiān)測(cè))。加入飽和碳酸氫鈉溶液(20 mL)，攪拌5 min后分液，有機(jī)相依次用鹽水洗三次，水洗兩次，水相用CH2Cl2(3×40 mL)萃取，合并有機(jī)相，用無(wú)水MgSO4干燥，減壓蒸除溶劑，剩余物經(jīng)柱色譜[洗脫劑A：V(乙酸乙酯) ∶V(石油醚)=1 ∶4]分離得無(wú)色油狀液體3 14.84 g,收率86%；1H NMR(CDCl3)δ: 1.26～1.31(s, 6H, CH3), 2.40(s, 3H, ArCH3), 3.69～3.73(m, 1H, 5-Ha), 3.91～4.01(m, 3H, 5-Hb, 1-H, 6-Ha), 4.20～4.26(m, 1H, 6-Hb), 7.25～7.32(d,J=8.2 Hz, 2H, ArH), 7.74～7.76(d,J=8.3 Hz, 2H, ArH);13C NMR(DMSO-d6)δ: 21.26(3-CH3), 26.35(3-CH3), 26.76(ArCH3), 53.07(C6), 66.08(C5), 74.84(C1), 109.09(C3), 128.27, 128.46, 135.82, 138.41(C in Ar); ESI-MSm/z: Calcd for [M+Na]+293.1, found 293.0; Anal.calcd for C13H18O4S: C 54.53, H 6.34; found: C 54.48, H 6.32。

(3) 4的合成

在反應(yīng)瓶中加入DMF 100 mL,312.38 g(43.29 mmol)和NaN33.90 g(60.00 mmol),攪拌下于105 ℃反應(yīng)4 h。減壓蒸除溶劑，剩余物重用乙酸乙酯溶解，用水洗滌三次，無(wú)水硫酸鎂干燥，減壓濃縮得漿狀物，經(jīng)柱色譜(洗脫劑A)分離得無(wú)色油狀液體4 5.99 g,收率88%;1H NMRδ: 1.26(s, 3H, CH3), 1.41(s, 3H, CH3), 3.23～3.33(m, 1H, 5-Ha), 3.43～3.49(m, 1H, 5-Hb), 3.61～3.69(m, 1H, 1-H), 3.94～3.98(m, 1H, 6-Ha), 4.22～4.28(m, 1H, 6-Hb);13C NMRδ: 26.55(CH3), 26.92(CH3), 52.29(C6), 65.95(C5), 74.53(C1), 109.05(C3); ESI-MSm/z: Calcd for [M+Na]+180.1, found 180.0; Anal.calcd for C6H11N3O2: C 45.85, H 7.05, N 26.74; found: C 45.79, H 7.09, N 26.79。

山洪災(zāi)害問(wèn)題已經(jīng)引起世界各國(guó)和組織的普遍關(guān)注，近年，在山洪災(zāi)害防治方面所開(kāi)展的工作不斷深入，在科研、示范、監(jiān)測(cè)預(yù)警系統(tǒng)建設(shè)、風(fēng)險(xiǎn)管理等方面均取得了很大進(jìn)展。借鑒歐美等發(fā)達(dá)國(guó)家的經(jīng)驗(yàn)，結(jié)合我國(guó)山洪災(zāi)害防治工作現(xiàn)狀和實(shí)際需求，建議在今后繼續(xù)組織開(kāi)展如下幾個(gè)方面的工作，把我國(guó)山洪災(zāi)害防御工作推向一個(gè)新的高度。

(4)5的合成

在反應(yīng)管中加入H2O/t-BuOH 1 mL和4 1 mmol,搖勻后加入銅粉(4 mmol)和CuSO4·5H2O 0.2 mmol，搖勻形成懸浮液，于60 ℃在100 W超聲波下輻射20 min(TLC監(jiān)測(cè))。用乙酸乙酯(2×5 mL)萃取，合并有機(jī)相，用無(wú)水硫酸鎂干燥，濾液減壓蒸除溶劑，剩余物用柱色譜[洗脫劑A或洗脫劑B:V(甲醇) ∶V(二氯甲烷)=1 ∶19]分離得5,實(shí)驗(yàn)結(jié)果見(jiàn)表1。

5a:1H NMRδ: 1.36(s, 3H, CH3), 1.40(s, 3H, CH3), 3.75～3.79(m, 1H, 5-Ha), 4.12～4.15(m,

表 1 合成5的實(shí)驗(yàn)結(jié)果*Table 1 The experamental result of synthesizing 5

*洗脫劑A:V(乙酸乙酯) ∶V(石油醚)=1 ∶4,洗脫劑B:V(甲醇) ∶V(二氯甲烷)=1 ∶19,洗脫劑C:V(甲醇) ∶V(二氯甲烷)=1 ∶49;a括號(hào)內(nèi)數(shù)據(jù)對(duì)應(yīng)的反應(yīng)條件為：無(wú)超聲波輻射，于室溫反應(yīng)11 h，其余反應(yīng)條件同1.2(4)

1H, 5-Hb), 4.44～4.58(m, 2H, 1-H, 6-Ha), 4.61(dd,J=12.1 Hz, 2.2 Hz, 1H, 6-Hb), 7.31～7.35(m, 1H, ArH), 7.41～7.44(m, 2H, ArH), 7.83～7.85(m, 2H, ArH), 7.90(s, 1H, 7-H);13C NMRδ: 25.40(CH3), 26.82(CH3), 52.43(C6), 66.60(C5), 74.28(C1), 110.40(C3), 121.12(C7), 147.90(C8), 125.80, 128.31, 129.00, 130.74(C in Ar); Anal.calcd for C14H17N3O2: C 64.85, H 6.61, N 16.20; found C 64.90, H 6.59, N 16.17。

5b:1H NMR(DMSO-d6)δ: 1.26(s, 3H, CH3), 1.30(s, 3H, CH3), 3.77～3.80(m, 1H, 5-Ha), 4.07～4.11(m, 1H, 5-Hb), 4.46～4.53(m, 2H, 1-H, 6-Ha), 4.63(dd,J=12.1 Hz, 2.2 Hz, 1H, 6-Hb), 7.26～7.30(m, 2H, ArH), 7.88～7.92(m, 2H, ArH), 8.53(s, 1H, 7-H);13C NMR(DMSO-d6)δ: 25.12(CH3), 26.54(CH3), 52.00(C6), 65.90(C5), 73.82(C1), 109.17(C3), 122.20(C7), 145.34(C8), 115.90, 127.31, 160.54, 163.00(C in Ar); Anal.calcd for C14H16N3O2F: C 60.64, H 5.82, N 15.15; found C 60.70, H 5.83, N 15.19。

5c:1H NMRδ: 1.26(s, 3H, CH3), 1.30(s, 3H, CH3), 3.76～3.78(m, 1H, 5-Ha), 3.80(s, 3H, OCH3), 4.06～4.10(m, 1H, 5-Hb), 4.44～4.50(m, 2H, 1-H, 6-Ha), 4.56～4.59(dd,J=12.1 Hz, 2.2 Hz, 1H, 6-Hb), 6.99～7.03(m, 2H, ArH), 7.76～7.79(m, 2H, ArH), 8.42(s, 1H, 7-H);13C NMRδ: 25.24(CH3), 26.57(CH3), 52.08(C6), 55.12(OCH3), 65.96(C5), 73.87(C1), 109.10(C3), 121.31(C7), 146.14(C8), 114.34, 123.36, 126.50, 159.03(C in Ar); Anal.calcd for C15H19N3O4: C 62.27, H 6.62, N 14.52; found C 62.32, H 6.63, N 14.56。

5d:1H NMRδ: 1.26(s, 3H, CH3), 1.30(s, 3H, CH3), 2.33(s, 3H, ArCH3), 3.76～3.80(m, 1H, 5-Ha), 4.06～4.10(m, 1H, 5-Hb), 4.44～4.51(m, 2H, 1-H, 6-Ha), 4.57～4.60(dd,J=15.4 Hz, 1.5 Hz, 1H, 6-Hb), 7.24～7.26(m, 2H, ArH), 7.73～7.75(m, 2H, ArH), 8.46(s, 1H, 7-H);13C NMRδ: 20.81(ArCH3), 25.23(CH3), 26.51(CH3), 52.04(C6), 66.03(C5), 73.82(C1), 109.13(C3), 121.84(C7), 146.21(C8), 125.10, 128.04, 129.43, 137.26(C in Ar); Anal.calcd for C15H19N3O2: C 65.91, H 7.01, N 15.37; found C 65.87, H 7.03, N 15.40。

5e:1H NMRδ: 1.22(s, 3H, CH3), 1.23(s, 3H, CH3), 1.74(s, 3H, ArCH3), 3.68～3.72(dd,J=8.8 Hz, 5.4 Hz, 1H, 5-Ha), 4.02～4.06(dd,J=8.8 Hz, 6.2 Hz, 1H, 5-Hb), 4.41～4.47(m, 2H, 1-H, 6-Ha), 4.51～4.57(dd,J=16.2 Hz, 6.3 Hz, 1H, 6-Hb), 4.90(s, 2H, NCH2), 7.63(s, 1H, 6′-H), 8.03(s, 1H, 7-H), 11.33(s, 1H, NH);13C NMRδ: 12.06(ArCH3), 25.30(CH3), 26.52(CH3), 42.25(C6), 51.87(C5), 65.96(NCH2), 73.86(C1), 108.96(C5′), 109.18(C3), 124.61(C7), 141.25(C6′), 142.47(C8), 150.84(C2′) , 164.37(C4′); Anal.calcd for C14H19N5O4: C 52.33, H 5.96, N 21.79; found C 52.37, H 5.95, N 21.76。

(5)6的合成

在反應(yīng)瓶中加入5的THF(3 mL)溶液，2 mol·L-1鹽酸1 mL,攪拌下于室溫反應(yīng)(TLC監(jiān)測(cè))。減壓蒸除溶劑，剩余物用柱色譜(洗脫劑B)分離得6。實(shí)驗(yàn)結(jié)果見(jiàn)表2。

表 2 合成6的實(shí)驗(yàn)結(jié)果*Table 2 The experamental result of synthesizing 6

*Rf=0.32,洗脫劑：V(甲醇) ∶V(二氯甲烷)=1 ∶4

6a:1H NMRδ: 3.18～3.38(m, 1H, 3-Ha), 3.41～3.46(m, 1H, 3-Hb), 3.86～3.91(m, 1H, 2-H), 4.24～4.30(dd,J=13.8 Hz, 8.2 Hz, 1H, 1-Ha), 4.52～4.57(dd,J=13.8 Hz, 3.5 Hz, 1H, 1-Hb), 4.86(t,J=5.5 Hz, 1H, 3-OH), 5.19(d,J=5.4 Hz, 1H, 2-OH), 7.30～7.34(m, 1H, ArH), 7.42～7.46(m, 2H, ArH), 7.85～7.87(m, 2H, ArH), 8.48(s, 1H, 5-H);13C NMRδ: 53.12(C1), 63.36(C3), 70.41(C2), 122.22(C5), 147.08(C4), 125.13, 127.70, 128.94, 130.95(C in Ar); ESI-MSm/z: Calcd for [M+Na]+242.1, found 242.1; Anal.calcd for C11H13N3O2: C 60.26, H 5.98, N 19.17; found C 60.21, H 5.97, N 19.21。

6b:1H NMRδ: 3.36～3.38(m, 1H, 3-Ha), 3.41～3.46(m, 1H, 3-Hb), 3.86～3.90(m, 1H, 2-H), 4.24～4.29(dd,J=13.8 Hz, 8.2 Hz, 1H, 1-Ha), 4.52～4.57(dd,J=13.8 Hz, 3.8 Hz, 1H, 1-Hb), 4.86(t,J=5.5 Hz, 1H, 3-OH), 5.19(d,J=5.4 Hz, 1H, 2-OH), 7.25～7.31(m, 2H, ArH), 7.87～7.91(m, 2H, ArH), 8.48(s, 1H, 5-H);13C NMRδ: 53.12(C1), 63.36(C3), 70.48(C2), 122.22(C5), 15．10(C4), 115.73, 115.90, 127.13, 127.58, 160.56, 162.90(C in Ar); ESI-MSm/z: Calcd for [M+Na]+260.1, found 260.1; Anal.calcd for C11H12N3O2F: C 55.69, H 5.10, N 17.71; found C 55.63, H 5.11, N 17.73。

6c:1H NMR(DMSO-d6)δ: 3.33～3.37(dd,J=11.0 Hz, 6.4 Hz, 1H, 3-Ha), 3.41～3.46(dd,J=11.0 Hz, 5.1 Hz, 1H, 3-Hb), 3.78(s, 3 H, OCH3), 3.87～3.90(m, 1 H, 2-H), 4.22～4.28(dd,J=13.8 Hz, 8.1 Hz, 1H, 1-Ha), 4.50～4.55(dd,J=13.8 Hz, 3.5 Hz, 1H, 1-Hb), 4.87(t,J=5.5 Hz, 1H, 3-OH), 5.19(d,J=5.4 Hz, 1H, 2-OH), 6.98～7.02(m, 2H, ArH), 7.76～7.79(m, 2 H, ArH), 8.37(s, 1H, 5-H);13C NMRδ: 53.02(C1), 55.14(OCH3), 63.32(C3), 70.51(C2), 121.32(C5), 145.94(C4), 114.33, 123.68, 126.59, 158.97(C in Ar); ESI-MSm/z: Calcd for [M+Na]+272.1, found 272.1; Anal.calcd for C12H15N3O3: C 57.82, H 6.07, N 16.86; found C 57.86, H 6.08, N 16.88。

6d:1H NMRδ: 2.32(s, 3H, ArCH3), 3.34～3.37(m, 1H, 3-Ha), 3.41～3.45(m, 1H, 3-Hb), 3.87～3.89(m, 1H, 2-H), 4.23～4.28(dd,J=13.8 Hz, 8.2 Hz, 1H, 1-Ha), 4.50～4.55(dd,J=13.8 Hz, 3.8 Hz, 1H, 1-Hb), 4.86(t,J=5.5 Hz, 1H, 3-OH), 5.18(d,J=5.4 Hz, 1H, 2-OH), 7.23～7.25(m, 2H, ArH), 7.73～7.75(m, 2H, ArH), 8.41(s, 1H, 5-H);13C NMRδ: 20.82(ArCH3), 53.05(C1), 63.37(C3), 70.51(C2), 121.83(C5), 146.01(C4), 125.04, 128.27, 129.43, 136.90(C in Ar); ESI-MSm/z: Calcd for [M+Na]+256.1, found 256.0; Anal.calcd for C11H12N3O2F: C 55.69, H 5.10, N 17.71; found C 55.63, H 5.11, N 17.73。

6e:1H NMRδ: 1.75(s, 3H, ArCH3), 3.26～3.32(m, 1H, 3-Ha), 3.36～3.41(m, 1H, 3-Hb), 3.78～3.83(m, 1H, 2-H), 4.18～4.24(dd,J=13.8 Hz, 8.1 Hz, 1H, 1-Ha), 4.50～4.49(dd,J=13.8 Hz, 3.4 Hz, 1H, 1-Hb), 4.81～4.84(t,J=5.6 Hz, 1H, 3-OH), 4.89(s, 2H, NCH2), 5.12(d,J=5.4 Hz, 1H, 2-OH), 7.61(s, 1H, 6′-H), 8.00(s, 1H, 5-H), 11.28(s, 1H, NH);13C NMRδ: 12.06(ArCH3), 42.25(C1), 52.90(NCH2), 63.37(C4), 70.45(C1), 108.96(C5′), 124.47(C5), 141.25(C6′), 142.19(C5), 150.84(C2′), 164.37(C4′); ESI-MSm/z: Calcd for [M+Na]+304.1, found 304.0; Anal.calcd for C12H15N3O2: C 61.79, H 6.48, N 18.01; found C 61.84, H 6.49, N 17.97。

2 結(jié)果與討論

2.1 1,3-偶極環(huán)加成反應(yīng)

以4a為例,反應(yīng)條件同1.2(4),于60 ℃考察超聲反應(yīng)時(shí)間，反應(yīng)30 min考察超聲反應(yīng)溫度對(duì)1,3-偶極環(huán)加成反應(yīng)的影響,實(shí)驗(yàn)結(jié)果見(jiàn)表3。由表3可見(jiàn)，宜選擇于60 ℃反應(yīng)20 min。

表 3 反應(yīng)時(shí)間和反應(yīng)溫度對(duì)1,3-偶極環(huán)加成反應(yīng)的影響*Table 3 Effect of reaction time and termperature on 1,3-dipolar cycloaddition

*于60 ℃考察反應(yīng)時(shí)間;反應(yīng)30 min考察反應(yīng)溫度;其余反應(yīng)條件同1.2(4)

2.2 5的脫保護(hù)

我們?cè)褂么姿?水、三氟乙酸-水和氨基磺酸-甲醇等體系在加熱條件下進(jìn)行5的脫丙酮基反應(yīng)，TLC監(jiān)測(cè)表明副產(chǎn)物較多，不易分離，最后選擇2 mol·L-1鹽酸-THF體系在室溫下脫保護(hù)得6。

2.3 6的抗癌活性

6具有開(kāi)環(huán)核苷的類(lèi)似結(jié)構(gòu)，以三唑基代替了核酸堿基。對(duì)一系列癌細(xì)胞[human hepatoma cell line(HepG2), human adenocarcinoma cell lines(A549), human lung adenocarcinoma cells line(LAC) and human Cervical Carcinoma cell lines(Hela)]的抗癌活性評(píng)價(jià)結(jié)果表明，當(dāng)c(6)<100μmol·L-1時(shí)，均未顯示有意義的抗癌活性。

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