本發(fā)明涉及化學(xué)領(lǐng)域，尤其涉及一種鬼臼毒素4-OH衍生物的制備方法。

背景技術(shù)：

木質(zhì)素類廣泛存在于植物王國之中，作為二級代謝產(chǎn)物來源于苯丙素的氧化二聚，按照傳統(tǒng)的分類方法木質(zhì)素類化合物可以分成兩大類，一類是經(jīng)典木質(zhì)素類即分子內(nèi)含有β-β’(8-8’)連接的木質(zhì)素類化合物，另一類是新型木質(zhì)素類即結(jié)構(gòu)中不含β-β’連接的苯丙素的二聚體，其中經(jīng)典類木質(zhì)素可以進一步細分為芳基四氫萘類(aryltetralin lignin)、二芐基丁烷類(dibenzylbutane lignan)、芳基萘類(arylnaphthalene lignan)及二苯并環(huán)辛二烯類(dibenzocyclooctadiene lignan)，生物活性測試顯示木質(zhì)素類化合物具有非常好的藥用活性，主要包括抗腫瘤、抗感染、免疫抑制、心腦血管保護、抗氧化及抗病毒等，最為著名的木質(zhì)素類化合物當屬鬼臼毒素，早期臨床試驗發(fā)現(xiàn)鬼臼毒素確實對腫瘤的治療非常高效，但遺憾的是其毒副作用太大，從而限制了其作為抗腫瘤藥物使用，而鬼臼毒素4-OH進行衍生化后不僅提高了所需活性，而且毒副作用也大大降低。例如依托泊苷，替尼泊苷，NK-611等，[Botta,B.；Delle Monache,G.；Misiti,D.；Vitali,A.；Zappia,G.Curr.Med.Chem.2001,8,1363]。

其中鬼臼毒素4-OH(1-O-[2-脫氧-2-(二甲氨基)-4:6-O-亞乙基-D-吡喃葡萄糖基]-4'-去甲基-L-表鬼臼毒素)是一種活性非常好的抗腫瘤藥物，已經(jīng)處于臨床二期，具有非常好的藥用前景[Invest New Drugs,1998,16,319]，然而鬼臼毒素4-OH的衍生化是化學(xué)領(lǐng)域的難點，主要是由于該羥基處于富電子苯環(huán)的芐位，非?；顫?，目前只有少數(shù)幾位化學(xué)家完成了鬼臼毒素及其衍生物4-O-糖苷的合成[(a)Helv.Chim.Acta.1968,51,163,Helv.Chim.Acta.1968,51,1631.(b)Tetrahedron Lett.1991,32,1653(c)Tetrahedron Lett.1992,33,4831.(d)J.Org.Chem.1993,58,4175.(e)Eur.J.Med.Chem.2012,47,424.]，但這幾種方法均具有產(chǎn)率低，構(gòu)型不易控制，毒性較大等缺點，我們課題組在前期已經(jīng)探索了利用鬼臼毒素4-OH與糖基炔酯給體制備糖基4-O-糖苷的方法[Org.Lett.2016,18,1294]，但是此方法只能用于合成4-O-糖苷類似物，而對于4-位N,C-等其他取代則無能為力，因此本發(fā)明主要提供一種鬼臼毒素及其類似物4-位炔酯給體與親核試劑進行反應(yīng)制備4-O-糖苷以及NK-611的方法，該方法不僅能用于合成4-O-糖苷鍵，而對于其他4-OH被N,C-等取代的類似物也可以高效地合成。

技術(shù)實現(xiàn)要素：

本發(fā)明的目的在于提供一種鬼臼毒素4-OH衍生物的制備方法，解決了鬼臼毒素及其類似物4-OH被糖基、氨基、碳及烯烴等取代的化合物合成方法缺點大的問題。

所述鬼臼毒素4-OH全名為1-O-[2-脫氧-2-(二甲氨基)-4:6-O-亞乙基-D-吡喃葡萄糖基]-4'-去甲基-L-表鬼臼毒素。

本發(fā)明的制備方法可適用于鬼臼毒素4-OH衍生物的制備，且反應(yīng)條件溫和，綠色環(huán)保，產(chǎn)物的產(chǎn)率和純度均較高。

本發(fā)明是這樣實現(xiàn)的，它包含下列步驟：

將鬼臼毒素類糖基炔酯給體2和親核試劑3進行糖苷化反應(yīng)，即可制得鬼臼毒素4-OH衍生物1；

其中，R₁為本領(lǐng)域常用的羥基保護基，所述的保護基為芐基(Bn，Cbz)，叔丁基二甲基硅基(TBS),甲基的一種；

NuH為具有親核性的羥基，氨基，烷基，烯基或炔基中的一種，當NuH為糖基羥基時，所述的糖基包括β-D-葡萄糖基、α-D-葡萄糖基、β-D-半乳糖基、α-D-半乳糖基、β-D-甘露糖基、α-D-甘露糖基、β-D-木糖基、α-D-木糖基、β-D-2-氨基葡萄糖基、α-D-2-氨基葡萄糖基、α-L-鼠李糖基、β-L-鼠李糖基、α-D-核糖基、β-D-核糖基、α-L-核糖基、β-L-核糖基、α-D-阿拉伯糖基、β-D-阿拉伯糖基、α-L-阿拉伯糖基、β-L-阿拉伯糖基、α-L-巖藻糖基、β-L-巖藻糖基、β-D-葡萄糖醛酸基、α-D-葡萄糖醛酸基、β-D-半乳糖醛酸基、或者α-D-半乳糖醛酸基；

本發(fā)明中，所述的制備的鬼臼毒素4-OH衍生物的方法包含下列步驟：在有機溶劑中、在干燥劑存在下、惰性氣體保護下，在親炔基路易斯酸的作用下，將鬼臼毒素類糖基炔酯給體2和親核試劑3進行糖苷化反應(yīng)，直到反應(yīng)完全為止；

所述的有機溶劑為干燥的二氯甲烷、甲苯、硝基甲烷和乙腈中的一種或多種。

所述的親炔基路易斯酸為AuCl、AuCl₃、AuLOTf、AuLNTf₂、HgOTf和PtCl₂中的一種或多種，其中，L為本領(lǐng)域常規(guī)的膦配體。

所述的膦配體為三丁基膦、三乙基膦、三苯基膦或三金剛烷基膦。

所述的親炔基路易斯酸的用量為化合物2的摩爾量的0.2～0.3倍。

所述的糖苷化反應(yīng)的溫度為25～35℃。

所述的糖苷化反應(yīng)時間為1～4小時。

所述的干燥劑的用量為鬼臼毒素類糖基炔酯給體2的摩爾量的1.0～4.0倍；所述的干燥劑為分子篩、分子篩、分子篩、酸洗的分子篩、酸洗的分子篩、酸洗的分子篩、無水硫酸鈉、無水硫酸鈣、無水硫酸銅和無水硫酸鎂中的一種或多種。

本發(fā)明的技術(shù)效果是：可適用于鬼臼毒素4-OH衍生物的制備，且反應(yīng)條件溫和，綠色環(huán)保，產(chǎn)物的產(chǎn)率和純度均較高。

具體實施方式

下面將結(jié)合實施例1-16詳細說明本發(fā)明所具有的有益效果，旨在幫助閱讀者更好地理解本發(fā)明的實質(zhì)，但不能對本發(fā)明的實施和保護范圍構(gòu)成任何限定。

下述各實施例中涉及到的室溫均為20～35℃。

實施例1

表鬼臼毒素4-O(甲基2,3,4-三-O-苯甲?；?α-D-吡喃葡萄糖)表鬼臼毒素糖苷的制備，

步驟1:

鬼臼毒素4-O-鄰環(huán)丙基乙炔基苯甲酸酯給體的合成：

在氮氣保護下，將鬼臼毒素3.5g，8.5mmol及鄰炔基苯甲酸2.4g12.75mmol溶于干燥的10mLDCM溶液中,然后加入DMAP(1.58g，12.75mmol)和DCC(3.5g，17.02mmol)，室溫攪拌2小時，此時TLC顯示所有起始原料消失，加入DCM(40mL)稀釋反應(yīng)混合物，所得溶液依次用水和飽和鹽水洗滌，蒸發(fā)并減壓濃縮，得到殘余物，將其通過硅膠色譜法(洗脫劑系統(tǒng)：PE：EA＝3:1)進一步純化,得到呈白色固體的(4.8g，97％)，[α]_D²⁵＝-169.1(c 1,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.88(dd,J＝1.2,8.0Hz,1H),7.51-7.43(m,2H),7.35(dt,J＝1.6,7.6Hz,1H),6.95(s,1H),6.57(s,1H),6.46(s,2H),6.20(d,J＝8.0Hz,1H),5.98(dd,J＝1.6,3.6Hz,2H),4.65(d,J＝4.0Hz,1H),4.54(dd,J＝6.0,8.8Hz,1H),4.36(t,J＝8.8Hz,1H),3.79(s,3H),3.77(s,6H),3.12-3.06(m,1H),3.03(dd,J＝4.0,14.4Hz,1H),1.29-1.24(m,1H),0.85-0.79(m,2H),0.78-0.72(m,2H)；¹³C NMR(100MHz,CDCl₃)δ173.3,166.6,152.2,147.7,147.2,136.6,134.4,134.0,132.0,131.5,130.8,129.4,128.0,126.8,124.1,109.3,107.5,107.1,101.2,99.2,73.9,73.7,71.1,60.3,55.7,45.2,43.4,38.3,8.34；HRMS(ESI)calcd for C₃₄H₃₁O₉[M+H]⁺583.19626found 583.19679；

步驟2：

表鬼臼毒素4-O(甲基2,3,4-三-O-苯甲?；?α-D-吡喃葡萄糖)表鬼臼毒素糖苷的制備，

在氮氣保護下,將甲基2,3,4-三-O-苯甲酰基-α-D-吡喃葡萄糖基(30mg，0.05mmol)和鬼臼毒素4-O-鄰-環(huán)丙基乙炔基苯甲酸酯給體(30mg，0.06mmol)溶解于干燥的的CH₂Cl₂(2mL)溶液中并加入加入將所得混合物在室溫下攪拌30分鐘，然后加入Ph₃PAuNTf₂(11mg，0.015mmol)，在室溫下繼續(xù)攪拌4小時(直到通過TLC監(jiān)測)，將混合物過濾，將濾液在減壓下濃縮，得到殘余物，將其通過硅膠柱色譜法(石油醚/乙酸乙酯3:1)進一步純化，得到白色固體41mg，92％，[α]_D²⁵＝+7.0(c 1.05,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.90(dd,J＝1.2,8.0Hz,2H),7.87(dd,J＝1.2,8.0Hz,2H),7.78(dd,J＝1.2,8.0Hz,2H),7.47-7.39(m,2H),7.34-7.26(m,5H),7.21-7.17(m,2H),6.68(s,1H),6.42(s,1H),6.15(s,2H),6.09(t,J＝10.0Hz,1H),5.87(d,J＝1.6Hz,1H),5.81(d,J＝1.6Hz,1H),5.51(t,J＝9.6Hz,1H),5.20(dd,J＝3.6,10.0Hz,1H),5.14(d,J＝3.6Hz,1H),4.50(dd,J＝8.8,10.8Hz,1H),4.47(d,J＝1.6Hz,1H),4.36(d,J＝3.2Hz,1H),4.34(t,J＝8.0Hz,1H),4.12-4.08(m,1H),3.82(dd,J＝2.4,10.4Hz,1H),3.70(s,3H),3.64(s,6H),3.62(dd,J＝4.8,10.4Hz,1H),3.38(dd,J＝5.2,14.0Hz,1H),3.35(s,3H),2.85-2.76(m,1H)；¹³C NMR(100MHz,CDCl₃)δ173.9,164.7(2C),164.3,151.4,147.4,145.5,136.1,134.4,132.5,132.3,132.0,131.5,128.8,128.7,128.6,128.1,127.9,127.8,127.4(2C),127.3,127.2,110.0,108.4,107.2,100.4,96.0,74.4,70.9,69.3,68.6,68.0,66.5,59.6,55.2,54.6,43.0,39.9,37.3。

實施例2

表鬼臼毒素4-O(甲基2,3,4-三-O-苯甲?；?α-D-吡喃葡萄糖)表鬼臼毒素糖苷的制備，

步驟1：

如實施例1的步驟1所示，[α]_D²⁵＝-50.8(c 1,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.89(dd,J＝1.2,8.0Hz,1H),7.49(dd,J＝1.6,8.0Hz,1H),7.45(td,J＝1.6,7.2Hz,1H),7.33(td,J＝1.6,8.0Hz,1H),7.00(s,1H),6.58(s,1H),6.46(d,J＝3.6Hz,1H),6.32(s,2H),6.00(d,J＝1.2Hz,1H),5.96(d,J＝1.6Hz,1H),4.72(d,J＝5.2Hz,1H),4.45(dd,J＝8.0,8.8Hz,1H),4.17(dd,J＝8.8,10.8Hz,1H),3.82(s,3H),3.76(s,6H),3.53(dd,J＝4.8,14.0Hz,1H),3.14-3.07(m,1H),1.40-1.36(m,1H),0.92-0.89(m,2H),0.81-0.73(m,2H)；¹³C NMR(100MHz,CDCl₃)δ174.0,165.9,152.3,148.5,147.1,137.0,134.3(2C),132.6,131.7,130.4,130.2,127.5,126.9,124.1,109.7,109.5,107.8,101.2,99.3,76.8,74.3,68.6,67.4,60.4,55.9,43.6,41.5,36.6,8.54,8.50；HRMS(ESI)calcd for C₃₄H₃₁O₉[M+H]⁺583.19626found 583.19683；

步驟2：

如實施例1的步驟2所示。

實施例3

表鬼臼毒素4-O(甲基2,3,4-三-O-芐基-α-D-吡喃葡萄)表鬼臼毒素糖苷的制備，

步驟1：

同實施例1中的步驟2所示，得到淡黃色的表鬼臼毒素4-O(甲基2,3,4-三-O-芐基-α-D-吡喃葡萄)表鬼臼毒素糖苷41mg,收率95％，[α]_D²⁵＝-19.8(c 1,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.40-7.23(m,15H),6.81(s,1H),6.52(s,1H),6.22(s,2H),6.00(d,J＝1.6Hz,1H),5.95(d,J＝1.6Hz,1H),5.01(d,J＝10.8Hz,1H),4.90(d,J＝11.2Hz,1H),4.83(d,J＝12.0Hz,1H),4.82(d,J＝10.4Hz,1H),4.68(d,J＝12.4Hz,1H),4.57(d,J＝4.4Hz,1H),4.56(d,J＝1.2Hz,1H),4.55(d,J＝11.2Hz,1H),4.35(d,J＝3.2Hz,1H),4.32-4.24(m,2H),4.01(t,J＝9.2Hz,1H),3.79(s,3H),3.75(m,1H),3.73(s,6H),3.71(m,1H),3.61-3.51(m,1H),3.43-3.31(m,2H),3.34(s,3H),2.85-2.76(m,1H)；¹³C NMR(100MHz,CDCl₃)δ175.0,152.5,148.4,146.7,138.6,138.2,138.1,137.2,135.4,132.5,129.0,128.5(2C),128.4,128.2,128.1(2C),128.0(3C),127.9,127.7,127.6,110.7,109.8,108.3,101.5,98.0,82.0,80.2,77.8,75.8,75.4,74.9,73.5,70.6,69.5,67.5,60.8,56.3,55.3,43.9,41.0,38.5；HRMS(ESI)calcd for C₅₀H₅₂O₁₃Na[M+Na]⁺883.33001 found 883.33188。

實施例4

表鬼臼毒素4-O(甲基-2,3-二-O-苯甲?；?6-O-芐基-α-D-葡萄)表鬼臼毒素糖苷的制備，

步驟1、2同實施例1中步驟1和2所示，得到化合物41.2mg,93％的收率，[α]_D²⁵＝-25.3(c 1.2,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.90(m,2H),7.60(dd,J＝1.2,8.0Hz,2H),7.48-7.40(m,4H),7.37-7.24(m,7H),6.81(s,1H),6.03(s,2H),5.86(d,J＝1.2,1H),5.82(d,J＝1.6Hz,1H),5.69(dd,J＝9.2,10.0Hz,1H),5.60(s,1H),5.25(dd,J＝3.6,10.0Hz,1H),5.15(d,J＝4.0Hz,1H),5.02(d,J＝12.0Hz,1H),4.55(d,J＝12.0Hz,1H),4.34(t,J＝9.6Hz,1H),4.18-4.10(m,3H),3.90-3.87(m,1H),3.80-3.73(m,2H),3.77(s,3H),3.72(s,6H),3.68(dd,J＝2.4,11.2Hz,1H),3.44(dd,J＝5.6,14.0Hz,1H),3.41(s,3H),2.41-2.32(m,1H)；¹³C NMR(100MHz,CDCl₃)δ175.1,165.9,165.2,152.3,147.9,146.1,137.1,136.8,135.9,133.3,132.8,132.3,129.8,129.2(2C),129.1,129.0,128.6,128.5,128.4,127.9,111.0,109.5,108.4,101.2,97.2,74.2,71.8,71.4,71.3,71.1,70.3,67.2,66.8,60.7,56.3,55.5,43.7,39.6,38.1。

實施例5

1,2；5,6-二異亞丙基-3-O-鬼臼毒素-α-D-吡喃葡萄糖基和1,2；5,6-二異亞丙基-3-O-表鬼臼毒素-α-D-吡喃葡萄糖基的制備，

同實施例1中步驟1和2所示，得到兩種化合物PPT和EPPT62.3mg(PPT:EPPT＝1.6：1)[α]_D²⁵＝-80.3(c 1.26,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ6.87(s,1H),6.54(s,1H),6.25(s,2H),6.00(d,J＝1.6Hz,1H),5.99(d,J＝3.6Hz,1H),5.98(d,J＝3.6Hz,1H),4.60(d,J＝4.8Hz,1H),4.59(d,J＝3.6Hz,1H),4.54(d,J＝3.2Hz,1H),4.44(dd,J＝8.4,10.8Hz,1H),4.36(t,J＝8.0Hz,1H),4.27-4.22(m,2H),3.85(dd,J＝2.0,10.0Hz,1H),3.80(s,3H),3.74(s,6H),3.72-3.61(m,2H),3.45(dd,J＝5.2,14.0Hz,1H),2.91-2.82(m,1H)；1.49(s,3H),1.39(s,3H),1.36(s,3H),1.33(s,3H)；¹³C NMR(100MHz,CDCl₃)δ175.1,152.5,148.5,146.7,137.2,135.5,132.4,129.0,112.3,110.8,109.7,108.3,106.4,101.5,101.1,84.0,79.5,75.0(2C),71.9,70.7,67.5,60.7,56.3,43.9,40.9,38.4,27.2,26.5,24.0；HRMS(ESI)calcd for C₃₄H₄₁O₁₃[M+H]⁺657.25417 found 657.25374.For the polar EPPT-17:[α]_D²⁵＝-58.2(c 0.9,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ6.98(s,1H),6.55(s,1H),6.22(s,2H),6.00(d,J＝3.6Hz,1H),5.98(d,J＝1.2Hz,1H),5.90(d,J＝4.0Hz,1H),4.73(d,J＝2.8Hz,1H),4.61(d,J＝5.6Hz,1H),4.57(d,J＝3.6Hz,1H),4.39(t,J＝8.0Hz,1H),4.31(dd,J＝8.4,10.8Hz,1H),4.21(d,J＝2.8Hz,1H),4.13-4.06(m,2H),4.03(dd,J＝5.6,8.4Hz,1H),3.95(dd,J＝4.4,5.2Hz,1H),3.80(s,3H),3.74(s,6H),3.52(dd,J＝5.6,14.0Hz,1H),3.00-2.91(m,1H),1.52(s,3H),1.35(s,3H),1.32(s,3H),1.26(s,3H)；¹³C NMR(100MHz,CDCl₃)δ174.7,152.6,148.6,146.7,137.3,135.4,133.0,128.0,112.2,111.0,110.0,109.0,108.3,105.3,101.5,82.8,81.3,80.0,73.3,72.3,67.4,67.2,60.7,56.3,44.0,40.6,38.4,26.8,26.6,26.3,25.1,24.7；HRMS(ESI)calcd for C₃₄H₄₁O₁₃[M+H]⁺657.25417 found 657.25318。

實施例6

表鬼臼毒素4-O膽固醇的制備，

同實施例1中步驟1和2所示，得到化合物56.4mg，收率100％。[α]_D²⁵＝-56.8(c 1,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ6.84(s,1H),6.51(s,1H),6.27(s,2H),6.00(d,J＝1.2Hz,1H),5.95(d,J＝1.2Hz,1H),5.39(d,J＝5.2Hz,1H),4.65(d,J＝3.2Hz,1H),4.60(d,J＝5.2Hz,1H),4.40(t,J＝8.0Hz,1H),4.26(dd,J＝8.4,11.2Hz,1H),3.80(s,3H),3.73(s,6H),3.39(dd,J＝5.2,14.0Hz,1H),3.28-3.22(m,1H),2.88-2.80(m,1H),2.38-2.26(m,2H),2.07-1.79(m,5H),1.68-1.42(m,8H),1.37-1.22(m,5H),1.01(s,3H),0.93(d,J＝6.4Hz,3H),0.88(d,J＝1.6Hz,3H),0.86(d,J＝2.0Hz,3H),0.68(s,3H)；¹³C NMR(100MHz,CDCl₃)δ175.0,152.6,148.2,147.1,140.3,137.2,135.3,132.2,130.5,122.2,110.4,109.3,108.3,101.4,79.2,71.3,67.8,60.7,56.8,56.3,56.2,50.2,43.9,42.4,41.2,39.8,39.5,39.4,38.5,37.2,36.9,36.2,35.8,31.9(2C),29.7,29.4,28.2,28.0,24.3,23.8,22.8,22.6,21.1,19.4,18.7,11.9；HRMS(ESI)calcd for C₄₉H₆₇O₈[M+H]⁺783.48305 found 783.48235。

實施例7

表鬼臼毒素4-O-(2-O-苯甲酰基-3,4,6-三-O-芐基-α/β-D-葡萄糖)表鬼臼毒素糖苷的制備，

同實施例1中步驟1和2所示，得到化合物81mg收率85％(α：β＝6:1)，[α]_D²⁵＝+54.2(c 0.63,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ8.00(dd,J＝1.2,8.0Hz,2H),7.62-7.57(m,1H),7.48(t,J＝7.6Hz,2H),7.42-7.31(m,5H),7.27-7.25(m,3H),7.20-7.07(m,7H),6.49(s,1H),6.21(s,2H),5.96(d,J＝1.6Hz,1H),5.92(d,J＝1.2Hz,1H),5.34(d,J＝4.0Hz,1H),5.28(dd,J＝3.6,10.0Hz,1H),4.80-4.70(m,5H),4.63(d,J＝5.2Hz,1H),4.59(d,J＝12.0Hz,1H),4.46(d,J＝10.4Hz,1H),4.23(dd,J＝4.4,10.8Hz,1H),4.08(t,J＝9.6Hz,1H),4.01(t,J＝J＝8.0Hz,1H),3.84(t,J＝9.2Hz,1H),3.79(s,3H),3.71(s,6H),3.68-3.60(m,3H),3.45(dd,J＝5.6,14.0Hz,1H),2.86-2.77(m,1H)；¹³C NMR(100MHz,CDCl₃)δ174.6,165.7,152.6,148.5,147.0,137.9(2C),137.6,137.3,135.1,133.8,132.5,129.6,129.0,128.9,128.8,128.5,128.4,128.3,128.2,128.0,127.9,127.8(2C),127.7,110.6,110.3,108.3,101.5,98.6,79.7,77.9,76.0,75.6,75.5,73.8,73.7,71.4,68.0,66.6,60.7,56.3,43.9,40.9,38.3；HRMS(ESI)calcd for C₅₆H₅₄O₁₄Cl[M+Cl]^-985.31966 found 985.32226。

實施例8

表鬼臼毒素-2-O-苯甲?；?3-O-芐基-4,6-O-亞芐基-α/β-D-葡萄糖苷的制備，

同實施例1中步驟1和2所示，得到化合物70.3mg，收率82％(α：β＝1.25:1)，[α]_D²⁵＝+32.9(c 0.87,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.99(dd,J＝1.2,8.4Hz,2H),7.79(dd,J＝1.2,8.4Hz,1.6H),7.64-7.36(m,14.4H),7.23-7.09(m,9H),6.94(s,1H),6.66(s,0.8H),6.52(s,1H),6.27(s,0.8H),6.21(s,2H),6.13(s,1.6H),5.984(d,J＝1.6Hz,1H),5.976(d,J＝1.2Hz,1H),5.87(d,J＝1.6Hz,0.8H),5.69(d,J＝1.6Hz,0.8H),5.64(s,0.8H),5.60(s,1H),5.40(d,J＝3.6Hz,1H),5.26(dd,J＝7.2,9.2Hz,1H),5.25(dd,J＝4.0,9.2Hz,0.8H),4.89-4.80(m,3H),4.75-4.69(m,2.8H),4.66(d,J＝5.2Hz,1H),4.47-4.39(m,1.6H),4.32(d,J＝5.2Hz,0.8H),4.26(t,J＝8.0Hz,0.8H),4.20-4.06(m,3H),3.97(t,J＝8.0Hz,1H),3.91-3.79(m,4H),3.78(s,3H),3.76(s,2.4H),3.71(s,6H),3.69(s,4.8H),3.58-3.50(m,1H),3.42(dd,J＝5.2,14.0Hz,1H),3.16(dd,J＝5.6,14.0Hz,0.8H),2.86-2.76(m,1.8H)；¹³C NMR(100MHz,CDCl₃)δ174.5(2C),165.8,164.9,152.6,152.5,148.6,148.4,146.9,146.6,137.9,137.8,137.3,137.2,137.1,137.0,135.2,135.1,133.9,132.9,132.6,129.7,129.6,129.5,129.2,129.1,128.8(2C),128.3(3C),128.2(2C),128.0,127.9,127.8,127.7,127.6,126.0,125.9,110.8,110.5,109.9,108.7,108.3,108.2,101.6,101.4,101.3(2C),100.0,99.1,82.2,81.7,75.4,74.8,73.9,73.8,73.4,68.6,68.5,67.6,66.5,66.4,63.1,60.7(2C),56.3,43.9,43.7,40.8,40.7,38.2,37.6；HRMS(ESI)calcd for C₄₉H₄₇O₁₄[M+H]⁺859.29603 found 859.29484。

實施例9

表鬼臼毒素-4-O-金剛烷基的制備，

同實施例1中步驟1和2所示，得到化合物44.4mg,收率81％，[α]_D²⁵＝-53.2(c 1.26,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ6.96(s,1H),6.48(s,1H),6.25(s,2H),5.98(d,J＝1.6Hz,1H),5.94(d,J＝1.2Hz,1H),4.93(d,J＝3.6Hz,1H),4.59(d,J＝5.6Hz,1H),4.38(dd,J＝8.0,10.8Hz,1H),4.28(t,J＝8.0Hz,1H),3.80(s,3H),3.74(s,6H),3.41(dd,J＝5.2,13.6Hz,1H),2.84-2.75(m,1H),2.22(bs,3H),1.86(s,6H),1.71(q,J＝12.0Hz,6H)；¹³C NMR(100MHz,CDCl₃)δ175.3,152.5,147.7,146.9,137.2,135.6,132.6,131.9,110.4,109.6,108.4,101.3,74.7,69.0,64.8,60.7,56.3,43.9,43.4,41.1,38.5,36.2,30.8。

實施例10

甲基-N-二苯基亞甲基-L-(4-O-表鬼臼毒素)苯基丙氨酸的制備，

同實施例1中步驟1和2所示，得到化合物70.2mg,收率93％，[α]_D²⁵＝-124(c 1.26,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.62(d,J＝8.8Hz,2H),7.42-7.29(m,6H),7.02(d,J＝8.4Hz,2H),6.78(d,J＝8.8Hz,2H),6.70(d,J＝7.2Hz,2H),6.64(s,1H),6.57(s,1H),6.30(s,2H),5.96(d,J＝1.2Hz,1H),5.92(d,J＝1.2Hz,1H),5.40(d,J＝3.6Hz,1H),4.69(d,J＝5.2Hz,1H),4.35(t,J＝8.0Hz,1H),4.28(dd,J＝4.4,9.2Hz,1H),4.13(dd,J＝8.4,10.8Hz,1H),3.81(s,3H),3.74(s,9H),3.46(dd,J＝5.2,14.0Hz,1H),3.27(dd,J＝4.4,13.6Hz,1H),3.20(m,1H),3.08-3.00(m,1H)；¹³C NMR(100MHz,CDCl₃)δ174.5,172.2,170.9,157.0,152.7,148.8,147.2,139.3,137.3,136.0,134.8,132.4,132.2,131.7,131.3,130.4,130.0,128.8(2C),128.6,128.3,128.2,128.1,127.6,115.7,110.2,109.3,108.2,101.6,73.4,67.5,67.3,60.8,56.3,52.3,43.8,41.6,38.9,38.1；HRMS(ESI)calcd for C₄₅H₄₁NO₁₀Na[M+Na]⁺778.26227found778.26218。

實施例11

表鬼臼毒素4-O-雌酚酮的制備，

同實施例1中步驟1和2所示，得到化合物55.3mg,收率83％，[α]_D²⁵＝-42.2(c 1.1,CHCl₃)；¹H NMR(400MHz,CDCl₃)7.24-7.22(d,J＝8.4Hz,2H),6.76-6.72(m,2H),6.70-6.68(M,1H),6.57(s,1H),6.32(s,2H),5.98(d,J＝1.6 1Hz 1H),5.94(d,J＝1.6 1H),5.46(d,J＝3.6Hz 1H),4.7(d,J＝4.8Hz,1H),4.36-4.30(t,J＝8.0Hz,1H),4.18-4.10(dd,J＝8.4,10.8Hz,1H),3.8-3.78(t,4H),3.7(s,6H),3.48-3.42(dd,J＝4.8,10.0Hz,1H),3.1.-3.02(m,1H),2.92-2.88(m,2H),2.56-2.48(m,1H),2.44-2.38(m,1H),2.30-2.22(m,1H),2.20-1.94(m,4H),1.7-1.42(m,7H)；¹³C NMR(100MHz,CDCl₃)δ174.6,156.3,152.6,148.7,147.2,138.4,137.3,134.8,133.5,132.2,128.9,126.7,115.9,112.8,110.1,109.3,108.2,101.5,77.2,72.7,67.7,60.77,56.30,50.42,47.9,44.0,43.8,41.6,38.2,35.8,31.5,29.6,26.4,25.8,21.6,13.8。

實施例12

4-氟-苯胺-鬼臼毒素和4-氟-苯胺-表鬼臼毒素的制備，

同實施例1中步驟1和2所示，得到化合物50.7mg，收率100％,(EPPT/PPT＝5:1)。[α]_D²⁵＝-104.4(c 0.66,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.15(s,0.25H),6.96-6.89(m,2.5H),6.76(s,1H),6.71(dd,J＝4.0,8.8Hz,0.5H),6.53(s,0.25H),6.52(2 1H),6.50(dd,J＝4.0,8.8Hz,2H),6.38(s,0.5H),6.32(s,2H),5.96-5.93(m,2.5H),4.64-4.58(m,2.5H),4.40(t,J＝8.0Hz,1H),4.26(dd,J＝7.2,8.8Hz,0.25H),4.01(dd,J＝8.4,10.4Hz,1.25H),3.80(s,3.75H),3.77(s,1.5H),3.75(s,6H),3.18(dd,J＝4.8,14.0Hz,1H),3.04-2.95(m,1H),2.91(dd,J＝4.8,14.0Hz,0.25H),2.75-2.65(m,0.25H),1.26(s,1.25H)；¹³C NMR(100MHz,CDCl₃)δ174.7,174.3,157.3,155.0,152.6(2C),148.3,147.7,147.6,143.8,137.5,137.3,135.7,135.1,131.7,131.5,130.6,129.0,128.2,125.3,116.3,116.2,116.1,116.0,113.1,113.0,110.1,116.2,116.1,116.0,113.1,113.0,110.1,109.9,109.1,108.7,108.4,106.9,101.6,101.4,71.2,68.8,60.7,56.4,56.3,53.3,46.4,44.0,43.6,41.8,38.7。

實施例13

4-脫氧-4-烯丙基-表鬼臼毒素的制備，

步驟1：

在氮氣保護下，將乙烯基三甲基硅烷和鬼臼毒素4-O-鄰-環(huán)丙基乙炔基苯甲酸酯給體溶在干燥的CH₂Cl₂中，并且加入分子篩(酒精噴燈稍微除水即可)，攪拌30min后，加入PPh₃AuNTf₂，在常溫下反應(yīng)四個小時，TLC監(jiān)控，得到化合物37mg,收率84％，[α]_D²⁵＝-67.7(c 1,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ6.73(s,1H),6.47(s,1H),6.29(s,2H),5.95(d,J＝1.2Hz,1H),5.94(d,J＝1.2Hz,1H),5.84-5.78(m,1H),5.16-5.10(m,2H),4.57(d,J＝5.2Hz,1H),4.28-4.25(m,2H),3.80(s,3H),3.74(s,6H),3.31-3.26(m,1H),3.10(dd,J＝5.2,14.0Hz,1H),3.02-2.94(m,1H),2.62-2.54(m,1H),2.46-2.39(m,1H)；¹³C NMR(100MHz,CDCl₃)δ175.0,152.4,147.1,146.9,137.1,136.7,136.1,133.1,130.9,116.9,110.1,108.7,108.4,101.2,69.0,60.7,56.2,44.1,42.3,38.5,37.7,36.2；HRMS(ESI)calcd for C₂₅H₂₇O₇[M+H]⁺439.17513 found439.17474。

實施例14

4-脫氧-4-苯甲酰甲基表鬼臼毒素的制備，

步驟1：

同實施例13中步驟1所示，得到化合物51mg,收率100％，[α]_D²⁵＝-27.9(c 1.26,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.98(dd,J＝1.2,8.4Hz,2H),7.63-7.59(m,1H),7.51(t,J＝7.6Hz,2H),6.70(d,1H),6.50(d,1H),6.32(s,2H),5.96(d,J＝1.6Hz,1H),5.94(d,J＝1.2Hz,1H),4.60(d,J＝4.8Hz,1H),4.35(dd,J＝7.6,9.2Hz,1H),4.07(qd,J＝2.0,6.0Hz,1H),3.81(s,3H),3.76(s,6H),3.60-3.49(m,2H),3.24(dd,J＝2.0,19.2Hz,1H),3.14-3.04(m,1H),2.94(dd,J＝4.8,14.0Hz,1H)；¹³C NMR(100MHz,CDCl₃)δ198.0,174.8,152.5,147.5,147.1,137.1,135.9,135.8,133.9,133.0,131.1,128.9,128.0,110.2,108.4,108.3,101.4,70.2,60.8,56.2,44.1,42.9,41.9,35.2,34.2；HRMS(ESI)calcd for C₃₀H₂₉O₈[M+H]⁺517.18569 found 517.18507。

實施例15

4-脫氧-4-(2-甲酰乙基)表鬼臼毒素的制備，

步驟1：

同實施例13中步驟1所示，得到化合物18mg,收率82％，[α]_D²⁵＝-55.5(c 0.64,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ9.80(s,1H),6.63(s1H),6.47δ6.28(s,2H),5.95(d,J＝1.2Hz,1H),5.94(d,J＝1.2Hz,1H),4.56(d,1H),4.34-4.28(dd,1H),3.88-3.82(m,1H),3.74(s,6H),3.62-3.56(dd,J＝9.2,11.6 1H),3.10-2.97(m,2H),2.85-2.75(m,2H)；¹³C NMR(100MHz,CDCl₃)δ199.5,174.5,152.5,147.5,147.1,137.2,135.6,132.2,131.0,110.2,108.3,108.0,101.4,77.3,77.2,77.0,76.7,69.7,60.7,56.45,48.09,44.483,41.92,34.92,32.89。

實施例16

鬼臼毒素4-OH的制備，

步驟1：化合物所需糖基受體的制備，

(1)

在氮氣保護下將異頭位TBS保護，二位疊氮保護，其他位置羥基裸露的β-D-葡萄糖500mg,1.57mmol和對甲苯磺酸27mg,0.16mmol溶解在10ml干燥的乙腈溶液中，攪拌30min后，加入0.5mL,4.71mmol二甲氧基乙烷，常溫下反應(yīng)4h,TLC監(jiān)控原料反應(yīng)完，用40ml乙酸乙酯稀釋，用飽和NaHCO₃洗，水洗，飽和食鹽水洗，無水硫酸鈉干燥，減壓濃縮柱層析(PE:EA＝4:1)得化合物471mg，收率87％。[α]_D²⁵＝-7(c 1.26,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ4.60(q,J＝4.8Hz,1H),4.45(d,J＝7.6Hz,1H),3.98(dd,J＝5.2,10.4Hz,1H),3.43(d,J＝10.4Hz,1H),3.38(d,J＝9.2Hz,1H),3.22(t,J＝9.2Hz,1H),3.15-3.09(m,3H),1.23(d,J＝5.2Hz,3H),0.77(s,9H),0.00(s,3H),-.002(s,3H)；¹³C NMR(100MHz,CDCl₃)δ99.8,97.5,80.2,71.5,69.1,68.0,66.3,25.5,20.2,17.9,-4.4,-5.2；HRMS(ESI)calcd for C₁₄H₂₈N₃O₅Si[M+H]⁺346.17928 found 346.17955、

(2)

將(1)所得產(chǎn)物56mg,0.16mmol和AZMBOH57mg,0.32mmol溶解在3ml干燥的DCM中，然后加入DMAP39mg,0.32mmol和DCC66mg,0.32mmol，常溫下反應(yīng)4h,TLC監(jiān)控原料反應(yīng)完，用40ml乙酸乙酯稀釋，用飽和NaHCO₃洗，水洗，飽和食鹽水洗，無水硫酸鈉干燥，減壓濃縮柱層析(PE:EA＝6:1)得化合物71mg，收率88％。[α]_D²⁵＝-21.3(c 1,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.84(dd,J＝1.6,8.0Hz,1H),7.43(td,J＝1.2,7.6Hz,1H),7.36(dd,J＝1.6,8.0Hz,1H),7.28(td,J＝1.6,7.6Hz,1H),5.10(t,J＝10.0Hz,1H),4.67-4.56(m,2H),4.52(q,J＝5.2Hz,1H),4.00(dd,J＝4.8,10.4Hz,1H),3.45-3.32(m,3H),3.26-3.19(m,1H),1.13(d,J＝5.2Hz,3H),0.76(s,9H),0.00(s,3H),-0.01(s,3H)；¹³C NMR(100MHz,CDCl₃)δ165.6,137.3,132.9,130.8,129.8,128.7,128.2,99.8,97.7,78.4,71.8,68.0,67.2,66.6,53.5,52.9,25.5,20.3,17.9,-4.4,-5.2；HRMS(ESI)calcd C₂₂H₃₃N₆O₆Si[M+H]⁺505.22254 found 505.22266、

(3)

將(2)所得產(chǎn)物128.4mg,0.255mmol,溶解在2ml干燥的吡啶中，反應(yīng)在塑料反應(yīng)器里面進行，在0℃條件下緩慢滴加HF-Pyr溶液0.32mL,3.57mmol，在0℃條件下攪拌3h,TLC監(jiān)控原料反應(yīng)完全。用40ml乙酸乙酯稀釋，用飽和1M HCl洗，飽和NaHCO₃洗，飽和食鹽水洗，無水硫酸鈉干燥，減壓濃縮柱層析(PE:EA＝3:1)得化合物94.5mg，收率95％。[α]_D²⁵＝+16.9(c 1,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ8.02(d,J＝4.4Hz,1H),8.00(d,J＝4.0Hz,1H),7.62-7.42(m,6H),5.81(t,J＝10.0Hz,1H),5.44(d,J＝3.2Hz,1H),5.34(t,J＝10.0Hz,1H),4.89(d.J＝8.0Hz,1H),4.85-4.70(m,6H),4.23(dd,J＝4.4,10.4Hz,1H),4.17-4.10(m,2H),3.63-3.45(m,6H),3.44(dd,J＝3.6,10.4Hz,1H),1.34(d,J＝4.8Hz,3H),1.32(d,J＝4.8Hz,3H)；¹³C NMR(100MHz,CDCl₃)δ166.1,165.7,137.3,133.0,130.8,129.8,128.6,128.1,99.9,96.8,93.2,79.1,78.2,72.0,69.7,68.3,68.0,66.0,65.7,62.6,62.2,52.7,20.2HRMS(ESI)calcd C₁₆H₁₈N₆O₆Na[M+Na]⁺413.11800 found 413.11818；

步驟2：

4'-脫甲基-4'-O-(芐氧羰基)表鬼臼毒素4-O-鄰-環(huán)丙基乙炔基苯甲酸酯

將4’去甲氧基表鬼臼毒素10mg,0.1mmol溶于干燥的DCM中，加入三乙胺0.02ml，0.15mmol,加入氯甲酸芐酯0.02ml,o.15mmol,室溫反應(yīng)3h,TLC監(jiān)控反應(yīng)結(jié)束，柱層析得到化合物，然后在氮氣保護下，將得到的化合物及鄰炔基苯甲酸516mg,1.87mmol溶于干燥的4mLDCM溶液中,然后加入DMAP(229mg,1.87mmol)和DCC(516mg,2.5mmol)，室溫攪拌2小時，此時TLC顯示所有起始原料消失，加入DCM(40mL)稀釋反應(yīng)混合物，所得溶液依次用水和飽和鹽水洗滌，減壓濃縮，得到殘余物，將其通過硅膠色譜法(洗脫劑系統(tǒng)：PE：EA＝4:1)進一步純化,得到呈白色固體的(577.6mg,66％兩步總收率)；

步驟3：

糖苷鍵的構(gòu)建

同實施例1中步驟2所示，得到化合物33.6mg,收率74％，[α]_D²⁵＝-17.2(c 0.53,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ8.00(dd,J＝1.6,8.0Hz,1H),7.62(td,J＝1.6,7.6Hz,1H),7.53(d,J＝7.6Hz,1H),7.46-7.32(m,6H),6.83(s,1H),6.57(s,1H),6.27(s,2H),6.03(d,J＝1.2Hz,1H),6.01(d,J＝1.2Hz,1H),5.29(t,J＝10.0Hz,1H),5.26(s,2H),5.00(d,J＝3.2Hz,1H),4.84-4.70(m,4H),4.68(d,J＝5.2Hz,1H),4.44(dd,J＝8.8,10.8Hz,1H),4.32-4.23(m,2H),3.67(s,6H),3.65-3.55(m,3H),3.49-3.43(m,1H),3.40(dd,J＝5.6,14.0Hz,1H),2.96-2.88(m,1H)；¹³C NMR(100MHz,CDCl₃)δ174.2,165.7,153.0,151.5,149.0,147.4,138.1,137.2,134.9,132.9,132.4,130.9,129.9,128.5,127.3,111.1,108.8,107.8,101.8,100.5,100.0,77.1,73.5,71.7,70.3,68.0,67.4,66.6,65.0,56.2,43.8,40.9,37.5,20.2HRMS(ESI)calcd C₄₅H₄₃N₆O₁₅[M+H]⁺907.27809 found 907.27910；

步驟4：

保護基的脫除，

將化合物4'-脫甲基-4'-O-芐氧羰基)表鬼臼毒素4-O-2“-脫氧-2”-疊氮基-3“-O-(疊氮甲基)苯甲酰-4'，6”-二-O-亞乙基-β-D-吡喃葡萄糖苷20mg溶解在2ml的甲醇溶液中，然后加入20mgPd(OH)₂/C,在反應(yīng)容器浸沒后，將容器抽真空，然后用H₂再填充,這個過程重復(fù)三次，然后在氫氣保護下反應(yīng)12h，TLC檢測，得到化合物10.9mg,收率85％，[α]_D²⁵＝-78.4(c 0.5,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ¹H NMR(400MHz,C₅D₅N)δ7.33(s,1H),6.8(s,1H),6.78(s,2H),5.96(s,1H),5.71(s,1H),5.06-5.02(m,4H),4.91-4.86(m,2H),4.72-4.66(m,1H),4.38-4.32(m,2H),4.09-4.07(t,J＝9.2Hz 1H),3.77(s,6H),3.75-3.59(m,5H),3.30-3.22(m 2H),1.45-1.40(d,J＝4.80H),¹³C NMR(100MHz,C₅D₅N)δ175.1,148.3,147.5,137.1,133.3,130.1,110.6,109.0,105.2,101.9,99.8,82.0,74.54,73.45,68.28,67.1,59.1,56.22,44.1,41.8,38.0,20.4HRMS(ESI)calcd C₂₉H₃₄NO₁₂[M+H]⁺588.20755 found 588.20679；

步驟5：

鬼臼毒素4-OH的制備，

操作步驟：在氮氣保護下，將4'-脫甲基-表鬼臼毒素4-O-2“-脫氧-2”-氨基-4“，6”-二-O-亞乙基-β-D-吡喃葡萄糖苷56mg溶解在2ml甲醇溶液中，加入20mg氰基硼氫化鈉，然后再加入甲醛水溶液0.084ml，之后反應(yīng)體系在常溫下攪拌4h，TLC監(jiān)控，反應(yīng)結(jié)束后，減壓濃縮，柱層析(CH₂Cl₂:MeOH＝20:1)得到化合物52mg,收率84％。[α]_D²⁵＝-32.3(c 1.26,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ6.73(s,1H),6.47(s,1H),6.29(s,2H),5.95(d,J＝1.2Hz,1H),5.94(d,J＝1.2Hz,1H),5.42(s,1H),5.0(d,J＝2.8Hz 1H),4.9-4.86(d,J＝8.4Hz,1H),4.81-4.76(dd,J＝4.8,10.0Hz 1H),4.66-4.63(d,J＝5.6Hz 1H),4.40-4.34(dd,J＝8.8,10.4Hz 1H),4.3-4.24(t,J＝8.0,8.4 1H),4.22-4.16(dd,J＝4.8,10.4Hz,1H),3.76(s,1H),3.68-3.58(m,2H),3.44-3.30(m,3H),2.95-2.87(m 1H),2.52-2.44(m 1H)2.32(s 6H)；¹³C NMR(100MHz,CDCl₃)δ174.6,148.8,147.1,146.5,134.3,133.1,130.5,127.6,111.2,108.8,107.7,101.7,99.84,97.8,80.77,77.0,71.4,68.1,67.6,66.5,56.4,43.7,41.1,37.4,20.4；HRMS(ESI)calcd C₃₁H₃₇NO₁₂Na[M+Na]⁺638.22080 found 638.21931。

以上所述的實施例僅僅是對本發(fā)明的優(yōu)選實施方式進行描述，并非對本發(fā)明的范圍進行限定，在不脫離本發(fā)明設(shè)計精神的前提下，本領(lǐng)域普通技術(shù)人員對本發(fā)明的技術(shù)方案作出的各種變形和改進，均應(yīng)落入本發(fā)明權(quán)利要求書確定的保護范圍內(nèi)。