CN102731610B - 26 sulfo-s or seleno spirostanol saponin, its synthetic method and application - Google Patents

Abstract

The present invention relates to a kind of 26 sulfo-s or seleno spirostanol saponin and chemical synthesis process thereof, the method comprises the steps: (1) 26 sulphonates of pseudo-spirostanol saponin unit and the nucleophilic reagent of sulphur or selenium react, and obtain 26 S or Se substitution products; (2) again with alkali, or reductive agent and acid-respons obtain 26 sulfo-s or seleno spirostanol saponin unit; (3) continue with acyl group, silica-based protection rhamnosyl three chlorimide ester to precursor reactant, obtain chacotriose sulphur glycosides, then through hydrolysis, with Trichloroacetonitrile react be converted into chacotriose three chlorimide esters to body; (4) react with the glycosyl donor of acyl group, alkyl and silica-based protection again, at the chacotriose that its 3 OH introduce full guard, obtain 26 sulfo-s of the full guard of α, β two kinds of configurations or the spirostanol saponin of seleno; (5) slough all protecting groups and obtain 26 sulfo-s or seleno spirostanol saponin.This compounds has stronger anti-tumor activity, can be used for the preparation of antitumor drug.

Description

26-position thio or selenospirosterol saponin, its synthesis method and application

technical field

The invention belongs to the field of drug synthesis, and relates to a thio or selenospirosteroid saponin, a chemical synthesis method thereof and an antitumor application.

technical background

Spirosteroid saponins are widely distributed in plants and have various biological activities: antitumor, antiviral, antibacterial, anti-inflammatory, and immune-stimulating activities. Dioscin is one of the most representative spirostanoid saponins, and has become the most widely used medical saponin [Viviane, SP; Alemandre, TC; Taketa, GG J.Braz.Chem.Soc. 2002, 13 , 135– 139]. A large number of experiments have shown that the activity of spirosterol saponins is determined by the sugar chains and aglycones, the number of sugar chains, the type of sugar groups, the stereo configuration and position of glycosidic bonds, and the isomerization and substitution of hydroxyl groups on sugar groups. And so on will have an important impact on the activity of saponins.

Sulfur and selenium, as the classic bioisosteres of oxygen, are often used in the structure optimization of lead compounds. In addition, organic sulfur and selenium compounds have attracted more and more attention in recent years because of their good biological activities[(a ) Yang Zhannan, Yang Xiaosheng Journal of Guizhou Normal University , 2004, 22 , 104–112; (b) Hu,C.;Zhang,P.;Li,Y.;Liu,B. Chemistry ,2002, 3 ,162–166; (c) Govindasamy, M.; Wolf-Walther, M.; Helmut, S.; Chem. Rev. 2001, 101 , 2125–2179]. Although there have been reports on the chemical synthesis of spirogenin substituted by sulfur or selenium [(a) Uhle, FC; J.Org.Chem. 1962, 27 , 2797–2799; (b) Quan, HJ; Koyanagi.J. ;Ohmori,K.;Uesato,S.;Tsuchido,T.;Saito,S. Eur.J.Med.Chem. 2002, 37 ,659–669], but no information about 26-position thio or selenium Report on the synthesis and application of spirulina saponins.

Contents of the invention

One of the objects of the present invention is to provide a 26-position thio or selenospironin synthesized from pseudospirostanoid sapogenin and potato triose.

Another object of the present invention is to provide a chemical synthesis method of the above-mentioned thio or selenospiroside, and its application in antitumor.

The 26-position thio or selenospirosteroid saponin structure of the present invention is as follows:

R ¹ -R ⁸ is any one of hydrogen, acyl or alkyl;

Potatotriose and its derivatives are linked with aglycon α or β glycosidic bonds;

The acyl group is a C ₂ -C ₆ linear or branched aliphatic acyl group or a C ₆ -C ₁₀ aromatic acyl group;

The alkyl group is methyl (Me), ethyl (Et), n-propyl ( n -Pr) or isopropyl ( i -Pr);

The absolute configuration of the 22-carbon of the spirosterol saponin is R , and the absolute configuration of the 25-carbon is R or S.

The synthetic method of described 26-position thio or selenospiroside comprises the steps:

(1) Synthesis of 26-position thio- or seleno-pseudospirogenin:

In a solvent or in the presence of a phase transfer catalyst, the 26-position sulfonate of pseudospironogenin reacts with a nucleophile of S or Se to obtain a corresponding 26-position S or Se-substituted product;

(2) Synthesis of 26-position thio or selenospirogenin:

Under the condition of 0-150°C, the 26-position S or Se substituted pseudospirogenin reacts with a base, or a reducing agent and an acid to obtain 26-position thio or selenospirogenin;

(3) Preparation of potato trisaccharide donor:

In the presence of a dehydrating agent, using Lewis acid or protonic acid as a promoter, 3,6-di-oxo-benzoyl-β-D-glucoglucoside and acyl, silicon-protected rhamnose trichloroimidate Donor reaction to obtain potato triose glucosinolate, which is then hydrolyzed and reacted with trichloroacetonitrile to be converted into a trichloroimidate donor of potato triose;

(4) Introduction of the 3-position sugar group:

In the presence of a dehydrating agent, with Lewis acid or protonic acid as a promoter, the 26-position thio or selenospironin reacts with the acyl or alkyl and silicon-protected sugar group donors, and the 3-position OH is introduced into the whole Protected potato triose, obtained fully protected 26-position thio- or seleno-spirosteroid saponins in two configurations of α and β;

(5) Synthesis of 26-position thio or selenospirosterol saponins:

In the presence of alkali, all the protecting groups of the fully protected spirostanol saponins are removed respectively to obtain 26-position thio or selenospirostanol saponins or derivatives thereof.

The specific steps are as follows:

(1) Synthesis of 26-position thio- or seleno-pseudospirogenin:

In a solvent or in the presence of a phase transfer catalyst, the sulfonate of 26-position pseudospirogenin reacts with a nucleophile of sulfur or selenium at 0-150°C for 1-10 hours to obtain 26-position S or Se substituted Product; Wherein, the mol ratio of sulfonic acid ester and nucleophile is 1.0:(1.0-5.0);

Described solvent is organic solvent, water or their mixture;

The organic solvent is C ₁ -C ₆ halogenated hydrocarbon, 1,4-dioxane, ether (Et ₂ O), acetonitrile (CH ₃ CN), 2,2,2-trimethylacetonitrile ( One of t -BuCN), tetrahydrofuran (THF), N , N -dimethylformamide (DMF), N , N -dimethylacetamide (DMA), hexamethylphosphoramide (HMPA), toluene or mixtures thereof;

The nucleophile is any one of KSCOCH ₃ , Na ₂ S ₂ , Li ₂ S ₂ , Cs ₂ S ₂ , K ₂ S ₂ or KSeCOCH ₃ , Na ₂ Se ₂ , Li ₂ Se ₂ , Cs ₂ Any one of Se ₂ , K ₂ Se ₂ ;

The phase transfer catalyst is tetrabutylammonium bromide (TBAB), tetrabutylammonium iodide (TBAI), tetrabutylammonium bisulfate (TBAH), 18-crown-6 (18-C-6), One of polyethylene glycol-400 (PEG-400).

(2) Preparation of 26-position thio or selenospirogenin:

In a solvent at 0-150°C, the above-mentioned S or Se substituted pseudospirogenin is hydrolyzed under the action of alkali and refluxed in an acidic solution, or reduced with a reducing agent in the presence of acetic acid for 0.1-10 hours to obtain the 26-position sulfur Generation or selenogenin; the molar ratio of the substitution product to the base or reducing agent is 1.0:(2.0-20.0):(2.0-5.0);

The base can be an inorganic base, such as one of sodium hydroxide, potassium hydroxide, lithium hydroxide, cesium hydroxide, sodium carbonate, potassium carbonate, and cesium carbonate; it can also be an organic base potassium tert-butoxide, methanol One of sodium, sodium ethoxide, and magnesium methoxide;

The acidic solution is 0.1-1mol/L hydrochloric acid methanol or ethanol solution.

(3) Preparation of potato trisaccharide donor:

In the presence of an organic solvent and a dehydrating agent, at -78-40 ° C, with Lewis acid or protonic acid as a promoter, 3,6-di-oxo-benzoyl-β-D-glucoglucoside and acyl, Silicon-protected rhamnose trichloroimidate donor is reacted for 1-10 hours to obtain potato triose glucosinolate, which is then converted into trisaccharide trichloroimidate donor by hydrolysis and reaction with trichloroacetonitrile;

The molar ratio of 3,6-di-oxy-benzoyl-β-D-glucosinolate to rhamnose donor and accelerator is 1.0:(1.0-5.0):(0.05-0.5); 3,6- The weight ratio of two-oxygen-benzoyl-β-D-glucosinolate and dehydrating agent is 1.0:(3.0-10.0);

The dehydrating agent is 3?, 4?, 5? molecular sieve or pickled 3? molecular sieve (AW-3?) or anhydrous sodium sulfate, anhydrous calcium sulfate, anhydrous copper sulfate, anhydrous magnesium sulfate one or a mixture of them;

The Lewis acid is C ₁ -C ₆ trialkylsilyl trifluoromethanesulfonate, boron trifluoride ether, silver trifluoromethanesulfonate, copper trifluoromethanesulfonate, zinc trifluoromethanesulfonate, Scandium trifluoromethanesulfonate, lanthanum trifluoromethanesulfonate, ytterbium trifluoromethanesulfonate, indium trifluoromethanesulfonate, trifluoromethanesulfonic acid, perchloric acid, tetrafluoroboric acid, tetrakis(pentafluorophenyl) One of boric acid or bis(trifluoromethanesulfonyl)imide;

The acyl group is C ₁ -C ₈ fatty acyl group, C ₆ -C ₁₀ aromatic acyl group, C ₇ -C ₁₀ aromatic halogenated hydrocarbon (such as benzyl chloride, p-methoxybenzyl chloride), and the silicon group is C ₁ -C ₆ trialkylsilyl.

(4) Introduction of 3-bit potato trisaccharides:

In an organic solvent and in the presence of a dehydrating agent, at -78-40 ° C, with a Lewis acid or a protonic acid as a promoter, 26-position thio- and selenogenin and acyl- or / and silicon-protected potato trichloride Imino ester donor reaction for 1-10 hours, introducing fully protected potato triose in its 3-OH, to obtain fully protected 26-position thio or selenospiroside in both α and β configurations;

The molar ratio of the 26-position thio or selenogenin to the glycosyl acceptor and accelerator is 1.0:(1.0-5.0):(0.05-0.5); the 26-position thio and selenogenin The weight ratio 1.0:(3.0-10.0) of element and dehydrating agent;

The dehydrating agent is 3?, 4?, 5? molecular sieve or pickled 3? molecular sieve (AW-3?) or anhydrous sodium sulfate, anhydrous calcium sulfate, anhydrous copper sulfate, anhydrous magnesium sulfate one or a mixture of them;

The Lewis acid is C ₁ -C ₆ trialkylsilyl trifluoromethanesulfonate, boron trifluoride ether, silver trifluoromethanesulfonate, copper trifluoromethanesulfonate, zinc trifluoromethanesulfonate, Scandium trifluoromethanesulfonate, lanthanum trifluoromethanesulfonate, ytterbium trifluoromethanesulfonate, indium trifluoromethanesulfonate, trifluoromethanesulfonic acid, perchloric acid, tetrafluoroboric acid, tetrakis(pentafluorophenyl) One of boric acid or bis(trifluoromethanesulfonyl)imide;

The glycosyl donor is a C ₁ -C ₈ linear or branched aliphatic acyl group, a C ₇ -C ₁₀ aromatic acyl group, and the silicon group is protected by a C ₁ -C ₆ trialkyl silicon group.

(5) Synthesis of 26-position thio or selenospiroside:

In a solvent, under the condition of -20-60°C, fully protected α and/or β two configurations of 26-position thio or selenospirosteroid saponins are reacted in the presence of alkali for 10-60 hours to remove all protection The 26-position thio or selenospirosterol saponin with two configurations of α and/or β; the base is an inorganic or organic base; the molar ratio of the fully protected spiroside to the base or hydrogenation catalyst is 1.0:(0.2-20);

The base can be an inorganic base, such as one of sodium hydroxide, potassium hydroxide, lithium hydroxide, cesium hydroxide, sodium carbonate, potassium carbonate, and cesium carbonate; it can also be an organic base potassium tert-butoxide, methanol One of sodium, sodium ethoxide, and magnesium methoxide.

The method of the present invention is recommended to be carried out under the protection of an inert gas, such as argon, nitrogen and the like.

The application of the 26-position thio or selenospirosteroid saponin in the preparation of antitumor drugs in the present invention.

The invention provides a simple and effective chemical method for synthesizing 26-position thio or selenospirosteroid saponins and their derivatives. These compounds have strong antitumor activity and can be used for the preparation of antitumor drugs.

Description of drawings

Fig. 1 is the synthesis process of 26-position thio or selenium diosgenin.

Fig. 2 is the synthesis process of diosgenin 13β, 14β and its isomers 13α and 14α at 26-position thio or selenium.

Detailed ways

Hereinafter, the present invention will be described in detail by taking the preparation and anti-tumor application of 26-position thio and selenium diosgenin and its α-isomer in conjunction with the accompanying drawings, but the present invention is not limited to the following content.

(1) Synthesis of 26-position thio or selenodiosgenin

As shown in Figure 1, the reagents and conditions in the synthesis reaction of 26-position thio or selenodiosgenin are: (a) KSCOCH ₃ , DMF, 60°C, 96%; (b) (i) KOH, MeOH, H ₂ O; (ii) HCl, EtOH, H ₂ O, reflux, 87%; (c) CsOH·H ₂ O, Se, N ₂ H ₄ ·H ₂ O, DMF, 60℃, 92%; (d ) (i) Zn, CH ₃ COOH, 150 ^o C; (ii) KOH, EtOH, H ₂ O, dioxane, 87%.

The specific experimental process and data are as follows:

Synthesis of Compound 2:

Under the protection of argon, the sulfonate 1 (369mg, 0.65mmol) [(a) Uhle, FC J.Org.Chem. 1962, 97 , 2797–2799; (b) Zha, X.; Sun, H.; Hao, J.; Zhang, Y. Chem. Biodiv . 2007, 4 , 25–31] was dissolved in 5 mL of DMF, and then KSCOCH ₃ (222 mg, 1.95 mmol, 3.0 equiv.) was added. After the reaction was stirred at room temperature for 10 h, TLC showed that the reaction was complete, concentrated under reduced pressure, diluted with CH ₂ Cl ₂ and washed with saturated NaCl, dried with Na ₂ SO ₄ , filtered, concentrated, and flash column chromatography to obtain the 26-position thioacetyl substituted Compound 2 (295 mg, 0.62 mmol, 96%).

[α] =-22.8( c 1.00, CHCl ₃ );

¹ HNMR(600MHz, CDCl ₃ ): δ 5.32(d,1H, J =5.4Hz),4.73-4.69(m,1H),3.51-3.47(m,1H),2.91(dd,1H, J =13.2, 5.4Hz),2.75(dd,1H, J =13.2,7.2Hz),2.44(d,1H, J =10.2Hz),2.30(s,3H),1.56(s,3H),1.00(s,3H) ,0.93(d,3H, J =6.6Hz),0.66(s,3H).

¹³ CNMR (150MHz, CDCl ₃ ): δ 195.9, 151.3, 140.8, 121.3, 103.7, 84.2, 71.5, 64.1, 54.9, 50.0, 43.2, 42.2, 39.4, 37.2, 36.5, 35.6, 34.0, 33.2, 32.8, 32.1, 31.5, 31.2, 30.6, 23.2, 20.9, 19.3, 18.9, 13.9, 11.6.

HRMS (ESI): [M+H] ⁺ , Calcd. for C ₂₉ H ₄₅ O ₃ S: 473.3084; Found: C ₂₉ H ₄₅ O ₃ S 473.3090.

Synthesis of compound 3:

Under argon protection, compound 2 (295 mg, 0.62 mmol) was dissolved in 20 ml of methanol, and then 5 mL of methanol/water (MeOH:H ₂ O=9:1) containing KOH (70 mg, 1.23 mmol, 2.0 equiv.) was added solution. After the mixture was stirred at room temperature for 15 min, TLC showed that the reaction was complete, then 6 mol/L hydrochloric acid aqueous solution (320 μL, 1.87 mmol, 3.0 equiv.) was added, and stirred at room temperature for 30 min, stored at 0°C for 12 h, and the solid was collected by filtration. After the solid was dried, it was dissolved in 20mL of ethanol/water (EtOH:H ₂ O=19:1) mixed solution under the protection of argon, then 0.5mL of 37% concentrated HCl was added, heated to reflux for 5h, cooled, poured Put into ice water, filter, collect the solid, and further purify by flash column chromatography to obtain 26-position S-substituted spirosteroidin 3 (209mg, 0.48mmol, 87%).

[α] =-162.7( c 1.10, CHCl ₃ );

¹ HNMR(600MHz, CDCl ₃ ): δ 5.34(d,1H, J =4,8Hz),4.62(dd,1H, J =15.0,7.2Hz),3.55-3.48(m,1H),2.52(t, 1H, J =13.2Hz), 2.28(d, 2H, J =13.2Hz), 2.22(t, 1H, J =12.6Hz), 1.01(s, 3H), 1.00(d, 3H, J =8.4Hz) ,0.92(d,3H, J =6.6Hz),0.80(s,3H).

¹³ CNMR (150MHz, CDCl ₃ ): δ 140.8, 121.4, 97.5, 81.6, 71.7, 62.8, 56.6, 50.0, 44.4, 42.2, 40.3, 39.7, 38.5, 37.2, 36.6, 33.3, 32.1, 32.0, 31.7, 31.6, 31.41, 31.38, 22.4, 20.8, 19.4, 16.5, 16.2.

HRMS (ESI): [M+H] ⁺ , Calcd. for C ₂₇ H ₄₃ O ₂ S: 431.2978; Found: C ₂₇ H ₄₃ O ₂ S 431.2983.

Synthesis of Compound 4:

Under argon protection, add CsOH·H ₂ O (76mg, 0.45mmol), selenium powder (23.7mg, 0.30mmol) into 2mL DMF, add dropwise N ₂ H ₄ ·H ₂ O (55μl, 0.90mmol) at room temperature , after stirring for 2h, sulfonate 1 (171mg, 0.30mmol) was added, then the temperature was raised to 60°C, stirring was continued for 4h, and then concentrated under reduced pressure. _The residue was diluted with _CH2Cl2 and washed with saturated NaCl. The organic phase was dried over Na ₂ SO ₄ , filtered, concentrated, and subjected to flash silica gel column chromatography to obtain diselenide 4 (132 mg, 0.14 mmol, 92%).

[α] =34.2( c 1.21, CHCl ₃ );

¹ HNMR(600MHz, CDCl ₃ ) δ 5.34(d,2H, J =4.8Hz),4.75-4.70(m,2H),3.53-3.49(m,2H),3.02(dd,2H, J =12.0,5.4 Hz),2.82(dd,2H, J =12.0,7.8Hz),2.45(d,2H, J =10.2Hz),2.30(m,4H),2.22(t,4H, J =10.8Hz)1.58(s ,6H), 1.01(s,6H), 0.99(d,6H, J =6.6Hz), 0.68(s,6H).

¹³ CNMR (150MHz, CDCl ₃ ) δ 151.5, 140.8, 121.3, 103.7, 84.3, 71.6, 64.2, 55.0, 50.0, 43.2, 42.2, 39.5, 39.1, 37.2, 36.6, 34.1, 33.9, 33.8, 32.2, 31.6, 3 , 23.4, 21.0, 19.5, 19.4, 14.0, 11.7.

Synthesis of Compound 5:

Under the protection of argon, 30 mL of acetic acid was added to the mixture of diselenide 4 (286 mg, 0.30 mol) and zinc powder (59 mg, 0.90 mmol, 3.0 eqiuv.), and heated to reflux for 24 h. TLC showed that the reaction was complete. The obtained residue was concentrated under reduced pressure, dissolved in 25 mL of dioxane under the protection of argon, then added to 20 mL of 10% KOH ethanol/water mixed solution (EtOH:H ₂ O=1:1), and stirred at room temperature for 24 h , the reaction solution was poured into ice water and extracted with CH ₂ Cl ₂ . The organic phase was washed with saturated NaCl, dried over Na ₂ SO ₄ , filtered, concentrated, and subjected to flash column chromatography to obtain compound 5 (250 mg, 0.52 mmol, 87%).

[α] =-192.8( c 1.00, CHCl ₃ );

¹ HNMR(600MHz, CDCl ₃ ): δ 5.34(d,1H, J =4.8Hz),4.64(dd,1H, J =15.6,7.8Hz),3.53-3.50(m,1H),2.58(t,1H , J =12.0Hz),2.37-2.34(m,1H),2.31-2.28(m,1H),2.25-2.20(m,2H),2.04-1.97(m,2H),1.02(d,3H, J =7.2Hz), 1.01(s,3H), 0.96(d,3H, J =6.6Hz), 0.80(s,3H).

¹³ CNMR (150MHz, CDCl ₃ ): δ 140.8, 121.4, 97.9, 82.7, 71.7, 62.9, 56.6, 50.0, 45.4, 42.2, 40.4, 40.1, 39.7, 37.2, 36.6, 34.0, 32.6, 32.0, 31.6, 31.4, 31.1, 24.8, 23.7, 20.8, 19.4, 17.7, 16.6.

(2) Synthesis of potato trisaccharide donor, 26-position thio or selenodioscin and its α-isomer

As shown in Figure 2, the reagents and conditions in the synthesis reaction of potato trisaccharide donor 10, 26-position thio or selenodioscin 13β, 14β and its isomers 13α and 14α are as follows: (a) TMSOTf, CH ₂ Cl ₂ , -30 ^o C; (b) NBS, acetone-H ₂ O, 72% (2-step reaction); (c) Cl ₃ CCN, DBU, CH ₂ Cl ₂ , 91%; (d) 3 or 5 ,4?MS,TMSOTf,CH ₂ Cl ₂ ,46%for11α; 27%for11β;,32%for12α;46%for12β;(e)CH ₃ ONa,CH ₃ OH:CH ₂ Cl ₂ =1:1,99 %for13α;95%for13β;90%for14α;78%for14β.

Synthesis of Compound 10:

Under argon protection, compound 6 [Song, G.; Yang, S.; Zhang, W.; Cao, Y.; Wang, P.; Ding, N.; Zhang, Z.; Guo, Y.; Li , Y. J.Med.Chem . 2009, 52 ,7368–7371.] ( _1.73g , _3.50mmol ), freshly activated 4? After stirring at ℃ for 30 min, TMSOTf (126 μL, 0.70 mmol) was added, and compound 7 was added after stirring for 10 min [(a) Kitagawa, I.; Back, NI; Ohashi, K.; Sakagami, M.; Yoshikawa, M.; Shibuya , Chem.Pharm.Bull. 1989, 37 ,1131–1133; (b) Cheng,MS;Wang,QL;Tian,Q.;Song,HY;Liu,YX;Li,Q.;Xu,X.;Miao , HD; Yao, XS; Yang, Z. J.Org.Chem. 2003, 68 ,3658–3662.] (4.56g, 10.49mmol) in CH ₂ Cl ₂ solution (5ml), stirred at -30°C for 2h After TLC detection, the reaction was complete, and the reaction was quenched by adding Et ₃ N, concentrated under reduced pressure, and subjected to flash column chromatography to obtain a mixture of compound 8 and compound 7 anomeric hydrolyzate.

Under the protection of argon, the product obtained in the previous step reaction was dissolved in 30mL acetone/water (9:1), and NBS (1.16g, 6.50mmol) was added under ice cooling, and after stirring for 30min, TLC showed that the reaction was complete. Saturated NaHCO ₃ quenched the reaction, concentrated under reduced pressure to remove acetone, and the residue was diluted with CH ₂ Cl ₂ , separated and collected the organic phase, then washed with saturated NaHCO ₃ and saturated NaCl successively, dried over anhydrous NaSO ₄ , filtered, and concentrated , and flash column chromatography to obtain compound 9 (2.34g, 2.51mmol, two-step total yield 72%).

Under the protection of argon, compound 9 (1.92g, 2.06mmol) was dissolved in CH ₂ Cl ₂ (25mL), DBU (125μL, 0.82mmol) was added under ice cooling, and after stirring for 5min, Cl ₃ CCN (2.06mL , 20.58mmol) to room temperature for 2h, TLC showed that the reaction was complete, and the reaction solution was concentrated under reduced pressure, followed by flash column chromatography to obtain compound 10 (2.02g, 1.87mmol, 91%).

[α] =103.2( c 1.05, CHCl ₃ );

¹ HNMR(600MHz, CDCl ₃ ): δ 8.74(s,1H),8.05(m,4H),7.59-7.54(m,2H),7.47-7.43(m,4H),6.48(d,1H, J = 3.6Hz),5.89(t,1H, J =10.2Hz),5.20(dd,1H, J =9.6,3.0Hz),5.15(dd,1H, J =3.6,1.8Hz),5.11(dd,1H, J =9.6,3.6Hz),4.93-4.87(m,3H),4.83-4.80(m,2H),4.79(d,1H, J =1.2Hz),4.51(dd,1H, J =12.6,3.6Hz ),4.33-4.30(m,1H),4.10(t,1H, J =9.6Hz),4.05(dd,1H, J =10.2,4.2Hz),3.90-3.87(m,1H),3.78-3.75( m,1H),2.00(s,3H),1.98(s,3H),1.95(s,3H),1.93(s,3H),1.86(s,3H),1.85(s,3H),1.13(d ,3H, J =6.6Hz),0.69(d,3H, J =6.0Hz).

¹³ CNMR (150MHz, CDCl ₃ ): δ 169.92, 169.88, 169.86, 169.82, 169.2, 169.0, 165.7, 165.1, 161.2, 133.2, 133.1, 129.9, 129.7, 129.6, 129.2, 128.3, 99.9, 9, 9 76.3, 72.3, 71.5, 70.8, 70.5, 69.9, 69.3, 68.4, 68.1, 67.5, 67.2, 62.0, 60.3, 20.65, 20.61, 20.45, 20.38, 17.2, 16.8, 14.1.

_HRMS (ESI): Calcd ^. for _{C46H52NO22Cl237Cl} : _1077.2012 ; Found: _1077.1991 .

Synthesis of Compound 11α, 11β:

Under the protection of argon, compound 3 (86 mg, 0.20 mmol) was dissolved in 4 mL of anhydrous CH ₂ Cl ₂ , and dry 4? Silyl trifluoromethanesulfonate (TMSOTf) (7μL, 0.04mmol, 0.2equiv.), continue to stir for 10min, then add ester-protected trisaccharide donor 10 (280mg, 0.26mmol, 1.30equiv.), -30 After stirring at ℃ for 4 hours, TLC showed that the reaction was complete, and the reaction was quenched with triethylamine. After filtration, concentration, and flash column chromatography, white solids 11α (125mg, 0.093mmol, 46%) and 11β (73mg, 0.054mmol, 27%) were obtained.

Compound 11α:

[α] =76.6( c 0.80, CHCl ₃ );

¹ HNMR(600MHz, CDCl ₃ ): δ 8.05-8.03(m,4H),7.56-7.51(m,2H),7.43-7.39(m,4H),5.77(t,1H, J =9.6Hz),5.25 (d,1H, J =4.2Hz),5.22(dd,1H, J =10.2,3.6Hz),5.17(dd,1H, J =10.2,3.6Hz),5.15-5.11(m,1H),5.05( d,1H, J =3.6Hz), 4.90(t,1H, J =9.6Hz), 4.89(t,1H, J =10.2Hz), 4.85(d,1H, J =1.8Hz), 4.77-4.72( m,3H),4.63(dd,1H, J =15.6,7.2Hz),4.52(dd,1H, J =12.6,5.4Hz),4.29-4.26(m,1H),3.91-3.86(m,2H) ,3.80-3.75(m,1H),3.72(dd,1H, J =9.6,3.0Hz),3.48-3.42(m,1H),2.53(t,1H, J =13.2Hz),2.47(d,2H , J =7.8Hz),2.28(d,1H, J =12.6Hz),2.01(s,3H),1.98(s,3H),1.95(s,3H),1.93(s,3H),1.87(s ,3H),1.85(s,3H),1.12(d,3H, J =6.0Hz),1.05(s,3H),1.00(d,3H, J =7.2Hz),0.92(d,3H, J = 6.6Hz), 0.80(s,3H), 0.69(d,3H, J =6.0Hz).

¹³ CNMR (150MHz, CDCl ₃ ): δ 170.0, 169.90, 169.87, 169.2, 165.9, 165.2, 140.1, 132.99, 132.96, 129.83, 129.77, 129.71, 129.6, 128.3, 1268.2, 1294.9, 9, 9.9 81.6,79.1,78.7,77.8,72.4,71.1,70.6,70.1,69.5,68.8,68.5,68.3,67.4,66.8,62.8,56.6,49.9,44.3,40.3,40.0,39.7,38.4,37.0,36.7,33.2, 32.0, 31.7, 31.4, 31.3, 27.8, 22.4, 20.73, 20.67, 20.5, 19.3, 17.4, 16.8, 16.5, 16.2.

HRMS (ESI): [M+Na] ⁺ , Calcd. for C ₇₁ H ₉₂ O ₂₃ SNa: 1367.5642; Found: 1367.5691.

Compound 11β:

[α] =34.6( c 0.80, CHCl ₃ );

¹ HNMR(600MHz, CDCl ₃ ): δ 8.04-8.01(m,4H),7.55-7.52(m,2H),7.44-7.38(m,4H),5.60(t,1H, J =9.4Hz),5.33 (d,1H, J =5.4Hz),5.14-5.11(m,2H),5.09(m,1H),4.95-4.93(m,1H),4.89(t,1H, J =10.4Hz),4.85( t,1H, J =10.2Hz),4.83(brs,1H),4.77(d,1H,J=11.52Hz),4.74(brs,1H),4.65(d,1H, J =7.68Hz),4.64- 4.60(m,1H),4.49(dd,1H, J =12.0,5.4Hz),4.35-4.30(m,1H),3.94(t,1H, J =9.6Hz),3.85-3.81(m,1H) ,3.77(t,1H, J =7.2Hz),3.70-3.66(m,1H),3.58-3.53(m,1H),2.52(t,1H, J =11.4Hz),2.38(d,1H, J =10.8Hz),2.28(d,1H, J =11.4Hz),2.23(t,1H, J =12.6Hz),1.96(s,6H),1.92(s,3H),1.90(s,3H), 1.86(s,3H),1.72(s,3H),1.13(d,3H, J =6.0Hz),1.00(d,3H, J =6.6Hz),0.93(s,3H),0.90(d,3H , J =6.0Hz), 0.78(s,3H),0.66(d,3H, J =6.0Hz).

¹³ CNMR (150MHz, CDCl ₃ ): δ 169.93, 169.88, 169.80, 169.6, 168.8, 165.7, 164.9, 140.0, 133.2, 132.9, 132.3, 130.8, 130.0, 129.8, 129.7, 129.1, 128.3, 128.3, 99.4,98.9,98.0,97.4,81.5,79.4,77.3,76.1,75.9,72.9,71.7,71.0,70.4,70.0,69.1,68.7,68.4,67.5,66.4,62.7,56.5,49.8,44.3,40.2,39.6, 38.45,38.36,36.8,36.7,33.2,32.02,31.99,31.7,31.3,29.6,27.6,22.4,20.73,20.68,20.65,20.62,20.5,20.2,19.1,17.1,16.8,16.5,16.1.

HRMS (ESI): [M+Na] ⁺ , Calcd. for C ₇₁ H ₉₂ O ₂₃ SNa: 1367.5642; Found: 1367.5696.

Synthesis of Compound 12α, 12β:

Under the protection of argon, compound 5 (81 mg, 0.17 mmol) was dissolved in 4 mL of anhydrous CH ₂ Cl ₂ , and dry 4? Silyl trifluoromethanesulfonate (TMSOTf) (6μL, 0.034mmol, 0.2equiv.), after stirring for 10min, was added ester-protected trisaccharide donor 10 (238mg, 0.22mmol, 1.30equiv.), -78 After stirring for 1 h at -40 °C for 3 h, TLC showed that the reaction was complete. After the reaction was quenched with triethylamine, it was filtered, concentrated, and flash column chromatography gave white solids 12α (76mg, 0.055mmol, 32%) and 12β (110mg, 0.079mmol, 46%).

Compound 12α:

[α] =-1.9( c 1.40, CHCl ₃ );

¹ HNMR(600MHz, CDCl ₃ ): δ 8.06(d,2H, J =4.2Hz),8.04(d,2H, J =4.2Hz),7.57-7.52(m,2H),7.44-7.40(dd,4H , J =13.8,7.8Hz),5.78(t,1H, J =9.6Hz),5.26(d,1H, J =4.2Hz),5.23(dd,1H, J =10.2,3.0Hz),5.18(dd ,1H, J =9.6,3.6Hz),5.15-5.12(m,1H),5.06(d,1H, J =3.6Hz),4.93-4.88(m,2H),4.86(brs,1H),4.78- 4.76(m,1H),4.74(d,1H, J =13.2Hz),4.66(dd,1H, J =15.6,7.2Hz),4.53(dd,1H, J =12.0,4.8Hz),4.32-4.26 (m,1H),3.91-3.89(m,2H),3.80-3.77(m,1H),3.73(dd,1H, J =9.6,3.6Hz),3.48-3.43(m,1H),2.59(t ,1H, J =12.0Hz),2.48(d,2H, J =7.8Hz),2.37(d,1H, J =11.4Hz),2.26-2.20(m,1H),2.02(s,3H),1.99 (s,3H),1.96(s,3H),1.94(s,3H),1.88(s,3H),1.86(s,3H),1.13(d,3H, J =6.0Hz),1.06(s, 3H), 1.02(d,3H, J =7.2Hz),0.96(d,3H, J =6.6Hz),0.81(s,3H),0.69(d,3H, J =6.0Hz).

¹³ CNMR (150MHz, CDCl ₃ ): δ 170.05, 169.93, 169.90, 169.2, 166.0, 165.2, 140.1, 133.02, 132.99, 129.88, 129.80, 129.76, 129.6, 128.34, 128.28, 990.9, 2, 9, 9 82.7,79.2,78.7,77.8,72.5,71.1,70.7,70.2,69.5,68.9,68.6,68.4,67.4,66.9,62.9,56.6,49.9,45.4,40.4,40.1,39.7,37.0,36.7,34.0,32.6, 32.0, 31.3, 31.1, 27.9, 24.8, 23.7, 20.8, 20.5, 19.3, 17.7, 17.4, 16.8, 16.6.

HRMS (ESI): [M+Na] ⁺ , Calcd. for _{C71H92O23SeNa} : _1415.5087 ; found: _1415.5119 .

Compound 12β:

[α] =-44.5( c 1.70, CHCl ₃ );

¹ HNMR(600MHz, CDCl ₃ ): δ 8.03(d,2H, J =8.4Hz),8.01(d,2H, J =8.4Hz),7.57-7.52(m,2H),7.45-7.39(m,4H ),5.60(t,1H, J =9.0Hz),5.33(d,1H, J =5.4Hz),5.15-5.12(m,2H),5.10-5.09(m,1H),4.95(dd,1H, J =3.6,1.8Hz),4.89(t,1H, J =9.6Hz),4.85(t,1H, J =9.6Hz),4.83(d,1H, J =1.8Hz),4.77(dd,1H, J =12.0,1.8Hz),4.74(d,1H, J =1.2Hz),4.65(d,1H, J =7.8Hz),4.64(dd,1H, J =12.4,7.8Hz),4.49(dd,1H , J =12.6,5.4Hz),4.34-4.31(m,1H),3.95(t,1H, J =9.6Hz),3.88-3.82(m,1H),3.77(t,1H, J =8.4Hz) ,3.71-3.67(m,1H),3.57-3.53(m,1H),2.57(t,1H, J =12.0Hz),2.39-2.34(m,2H),2.24-2.18(m,2H),1.96 (s,6H),1.93(s,3H),1.90(m,3H),1.86(s,3H),1.72(s,3H),1.13(d,3H, J =6.6Hz),1.01(d, 3H, J =6.6Hz),0.95(d,3H, J =7.8Hz),0.94(s,3H),0.78(s,3H),0.66(d,3H, J =6.6Hz).

¹³ CNMR (150MHz, CDCl ₃ ): δ 169.93,169.87,169.80,169.5,168.8,165.7,164.9,140.0,133.2,132.9,130.0,129.8,129.7,129.1,128.34,128.28,99,121.9 97.8,82.6,79.4,77.3,76.1,75.9,72.9,71.0,70.4,70.0,69.1,68.7,68.4,67.5,66.4,62.8,62.7,56.4,49.8,45.4,40.3,40.1,39.6,38.4,36.8, 36.7, 33.9, 32.6, 32.0, 31.3, 31.1, 29.6, 24.8, 23.6, 20.73, 20.67, 20.62, 20.5, 20.2, 19.1, 17.6, 17.1, 16.8, 16.5.

HRMS (ESI): [M+Na] ⁺ , Calcd. for C ₇₁ H ₉₂ O ₂₃ SeNa: 1415.5087; Found: 1415.5144.

Synthesis of Compound 13α:

Under argon protection, compound 11α (40mg, 0.0297mmol) was dissolved in a mixed solvent of 2ml of methanol/dichloromethane (MeOH:CH ₂ Cl ₂ =1:1), and then sodium methoxide (24mg, 0.44mmol, 14.6 equiv.) in methanol solution (230 μL), stirred at room temperature for 48 h, and TLC showed that the reaction was complete. The reaction was neutralized with an acidic resin and filtered to remove the solid. The filtrate was concentrated and flash column chromatographed to give 13α as a white solid (26 mg, 0.0294 mmol, 99%).

[α] =-46.0 ( c 0.09, CHCl ₃ :MeOH=1:1 containing 8% water);

¹ HNMR(600MHz,pyridine- d ₅ ): δ 5.89(s,1H),5.81(s,1H),5.46(d,1H, J =3.0Hz),5.27(d,1H, J =3.6Hz), 4.89-4.83(m,1H),4.81-4.76(m,2H),4.69-4.65(m,1H),4.62-4.55(m,2H),4.52-4.48(m,1H),4.42-4.37(m ,2H),4.33-4.28(m,2H),4.26(d,1H, J =10.2Hz),4.21(d,1H, J =12.0Hz),4.14-4.11(m,2H),3.73-3.68( m,1H),2.64-2.55(m,2H),2.49-2.42(m,1H),2.29(d,1H, J =10.8Hz),1.66(d,3H, J =6.0Hz),1.64(d ,3H, J =6.0Hz), 1.07(d,3H, J =6.0Hz),0.82(m,6H),0.78(s,3H).

¹³ CNMR(150MHz,pyridine- d ₅ ) δ 140.8,121.3,103.9,102.7,97.9,97.5,81.7,79.3,78.9,78.4,73.6,72.6,72.45,72.40,72.3,72.0,70.2,70.0,63.1,6 ,56.4,49.9,44.4,40.5,40.2,39.5,38.7,37.0,36.6,33.3,32.1,32.0,31.9,31.4,31.3,28.4,22.4,20.8,19.0,18.4,18.3,16.3.

HRMS (ESI): [M+Na] ⁺ , Calcd. for _C45H72O15SNa : _907.4484 ; found: _907.4505 .

Synthesis of Compound 13β:

According to the same method for compound 13α, compound 11β (40 mg, 0.0297 mmol) was deprotected to obtain white solid 13β (25 mg, 0.0282 mmol, 95%).

[α] =-97.4 ( c 0.85, CHCl ₃ : MeOH=1:1 containing 8% water);

¹ HNMR(600MHz,pyridine- d ₅ ): δ 6.36(s,1H),5.82(s,1H),5.27(s,1H),4.93-4.86(m,3H),4.79-4.75(m,2H) ,4.64(brs,1H),4.59-4.56(m,1H),4.51-4.48(m,1H),4.35-4.28(m,3H),4.19-4.17(m,3H),4.05(d,1H, J =12.6Hz),3.86-3.80(m,1H),3.60(d,1H, J =7.8Hz),2.74(d,1H, J =12.0Hz),2.68(t,1H, J =12.0Hz) ,2.60(t,1H, J =12.6Hz),2.30(d,1H, J =12.0Hz),1.72(d,3H, J =6.0Hz),1.58(d,3H, J =6.0Hz),1.07 (d, 3H, J =7.2Hz), 1.01(s, 3H), 0.79(m, 6H).

¹³ CNMR(150MHz,pyridine- d ₅ ): δ 140.5,121.5,102.6,101.8,100.0,97.5,81.7,78.2,77.8,77.7,77.5,76.7,73.8,73.6,72.6,72.4,72.2,70.1,69.2, 63.1, 61.0, 56.4, 50.0, 44.4, 40.2, 40.2, 39.5, 38.7, 37.2, 36.8, 33.3, 32.1, 32.0, 31.9, 31.45, 31.39, 29.9, 22.4, 20.8, 19.1, 18.4, 18.2, 16.3.

HRMS (ESI): [M+Na] ⁺ , Calcd. for C ₄₅ H ₇₂ O ₁₅ SNa: 907.4484; Found: 907.4501.

Synthesis of Compound 14α:

According to the same method of synthesizing compound 13α, white solid 14α (24 mg, 0.0258 mmol, 90%) was obtained from compound 12α (40 mg, 0.0287 mmol) through deprotection.

[α] =-87.4 ( c 0.85, CHCl ₃ : MeOH=1:1 containing 8% water);

¹ HNMR(600MHz,pyridine- d ₅ ) δ 5.92(s,1H),5.83(s,1H),5.48(d,1H, J =3.0Hz),5.27(d,1H, J =3.6Hz),4.91 -4.85(m,1H),4.81-4.76(m,2H),4.68-4.65(m,1H),4.62-4.57(m,2H),4.52(dd,1H, J =9.0,3.0Hz),4.42 -4.37(m,2H),4.33-4.28(m,2H),4.26(d,1H, J =10.2Hz),4.21(d,1H, J =12.0Hz),4.14-4.11(m,2H), 3.73-3.68(m,1H),2.66-2.60(m,2H),2.48(t,1H, J =11.4Hz),2.35(d,1H, J =12.0Hz),2.26-2.22(m,1H) ,2.14(d,1H, J =12.0Hz),2.00-1.97(m,1H),1.93-1.89(m,2H),1.66(d,3H, J =6.0Hz),1.64(d,3H, J =6.0Hz), 1.07(d,3H, J =6.0Hz), 0.82(m,6H), 0.78(s,3H).

¹³ CNMR(150MHz, Pyridine- d ₅ ): δ 140.8,121.3,103.9,102.7,98.0,97.9,82.8,79.3,79.0,78.4,73.6,72.6,72.5,72.4,72.3,72.0,70.2,70.1,63.2, 61.2, 56.4, 49.9, 45.4, 40.5, 40.3, 39.5, 37.0, 36.7, 34.0, 32.8, 32.0, 31.3, 31.1, 28.4, 24.7, 23.6, 20.8, 19.0, 18.4, 18.3, 17.7, 16.4.

HRMS (ESI): [M+Na] ⁺ , Calcd. for _{C45H72O15SeNa} : _955.3929 ; found: _955.3950 .

Synthesis of Compound 14β:

According to the same method of synthesizing compound 13α, white solid 14β (21 mg, 0.0225 mmol, 78%) was obtained from compound 12β (40 mg, 0.0287 mmol) through deprotection.

[α] =-148.7( c 0.80, CHCl ₃ :MeOH=1:1 containing 8% water);

¹ HNMR(600MHz,pyridine- d ₅ ) δ 6.40(s,1H),5.86(s,1H),5.28(d,1H, J =5.4Hz),4.97-4.92(m,3H),4.83-4.78( m,2H),4.67(brs,1H),4.62(dd,1H, J =9.0,3.6Hz),4.53(dd,1H, J =9.6,3.6Hz),4.41-4.30(m,3H),4.22 -4.18(m,3H),4.07(dd,1H, J =12.6,3.6Hz),3.88-3.83(m,1H),3.62(d,1H, J =9.6Hz),2.80-2.75(m,1H ),2.72-2.69(d,1H, J =11.4Hz),2.66(t,1H, J =10.8Hz),2.37(d,1H, J =11.4Hz),2.27-2.23(m,1H),2.04 -2.00(m,2H),1.75(d,3H, J =6.0Hz),1.62(d,3H, J =6.6Hz),1.07(d,3H, J =6.6Hz),1.02(s,3H) ,0.83(d,3H, J =6.6Hz),0.79(s,3H).

¹³ CNMR(150MHz,pyridine- d ₅ ) δ 140.5,121.5,102.6,101.8,100.0,97.8,82.8,78.2,77.7,77.6,77.5,76.6,73.8,73.6,72.5,72.4,72.2,70.1,62.2,63. ,61.0,56.3,50.0,45.4,40.3,39.4,38.6,37.2,36.8,34.0,32.8,32.0,31.3,31.2,29.9,24.7,23.6,20.8,19.1,18.4,18.2,17.7,16.4.

HRMS (ESI): [M+H] ⁺ , Calcd. for C ₄₅ H ₇₃ O ₁₅ Se: 933.4109; Found: 933.4137.

(3) Antitumor activity test of compounds

Compounds 13β, 13α, 14β and 14α and diosgenin were tested by MTT method [(a) Deng, S.; Yu, B.; Hui, Y. Tetrahedron Lett. 1998, 39 , 6511–6514; (b) Tao , HC; Yu, B. Tetrahedron Lett. 2001, 42 , 2405–2407] for the inhibitory activity of tumor cell lines A-549, Hela and K562, the results are shown in Table 1. Compared with diosgenin, the activity of the 26-position thio analogue 13β was increased, but the activity of the 26-position selenium analogue 14β was slightly lower than that of diosgenin, while the activities of 13α and 14α-type isomers were significantly decreased. This indicates that for spirosteroid saponins, the β-glycosidic bond between the 3-position sugar chain and the aglycone plays a key role in its antitumor activity.

Table 1: Antitumor activity (IC50) of 13α, 13β, 14α, 14β and diosgenin

compound A-549 Hela K562 Dioscin 4.02 7.86 5.12 13β 3.72 6.68 4.1 13α >10 >10 >10 14β 5.0 >10 4.96 14α >10 >10 >10

Claims (5)

1. A method for synthesizing 26-thio or seleno spirostanin with a structural general formula shown as the following,

wherein,

double bonds (Δ) in positions 5, 6⁵) (ii) a X = S or Se; y ═ H and H;

R¹-R⁸is hydrogen;

the potato trisaccharide and the derivatives thereof are connected with the aglycone by a beta glycosidic bond;

the absolute configuration of the carbon at each of the 22-and 25-positions isR；

The method is characterized by comprising the following steps:

(1) synthesizing 26-thio or seleno pseudo spirostanol sapogenin:

in a solvent or in the presence of a phase transfer catalyst, reacting 26-para-toluenesulfonate of the pseudo-spirostanol sapogenin with a sulfur or selenium nucleophile for 1 to 10 hours at the temperature of between 0 and 150 ℃ to obtain a corresponding sulfur or selenium substituted product; wherein the molar ratio of the p-toluenesulfonate to the nucleophile is 1 (1.0-5.0); the nucleophilic reagent is KSCOCH₃、Na₂S₂、Li₂S₂、Cs₂S₂、K₂S₂One of them or KSeCOCH₃、Na₂Se₂、Li₂Se₂、Cs₂Se₂、K₂Se₂One of (1); the phase transfer catalyst is one of tetrabutylammonium bromide, tetrabutylammonium iodide, tetrabutylammonium hydrogen sulfate and polyethylene glycol-400;

(2) preparing 26-thio or seleno spirostanol sapogenin:

reacting 26-thio-or seleno-pseudo-spirostanol sapogenin with alkali or a reducing agent in a solvent at 0-150 ℃, and then reacting with acid for 0.1-10 hours to obtain 26-thio-or seleno-spirostanol sapogenin; the molar ratio of the 26-position thio-or seleno-pseudo-spirostanol sapogenin to the alkali is 1.0 (2.0-20.0), and the molar ratio of the 26-position thio-or seleno-pseudo-spirostanol sapogenin to the reducing agent is 1.0 (2.0-5.0); the alkali is organic alkali or inorganic alkali; the acid is an organic acid or an inorganic acid;

(3) preparation of potato trisaccharide donor:

in the presence of an organic solvent and a dehydrating agent, 3, 6-di-oxygen-benzoyl-beta-D-glucosinolate reacts with acyl and silicon-based protected rhamnose trichloroimino ester donor for 1 to 10 hours at the temperature of between 78 ℃ below zero and 40 ℃ by taking Lewis acid or protonic acid as an accelerating agent to obtain potato trisaccharide glucosinolate, and then the potato trisaccharide glucosinolate is hydrolyzed and reacted with trichloroacetonitrile to be converted into trisaccharide trichloroimino ester donor; the mol ratio of the 3, 6-di-oxygen-benzoyl-beta-D-glucothioglycoside to the rhamnose donor and the accelerator is 1.0 (1.0-5.0) to 0.05-0.5; the weight ratio of the 3, 6-di-oxygen-benzoyl-beta-D-glucosinolate to the dehydrating agent is 1.0 (3.0-10.0);

(4) introduction of a 3-position sugar group:

in the presence of an organic solvent and a dehydrating agent, under the conditions of-78-40 ℃, Lewis acid or protonic acid is used as an accelerating agent, 26-position thio or seleno spirost sapogenin reacts with acyl and silicon-based protected glycosyl donor for 1-10 hours, and fully-protected potato trisaccharide is introduced into 3-position OH of the 26-position thio or seleno spirost sapogenin to obtain fully-protected spirost saponin with beta configuration; wherein, the molar ratio of the 26-thio or seleno spirostanol sapogenin to the glycosyl donor and the accelerator is 1.0 (1.0-5.0) to 0.05-0.5; the weight ratio of the 26 th thio or seleno spirostanol sapogenin to the dehydrating agent is 1.0 (3.0-10.0);

(5) synthesis of 26-thio or seleno spirosteroid saponin:

reacting the beta-configuration fully-protected spirostanin with alkali for 10-60 hours at-20-60 ℃ in a solvent, and removing all protecting groups to obtain the beta-configuration 26-thio-or seleno-spirostanin; the alkali is inorganic or organic alkali; the molar ratio of the fully-protected spirostanin to the alkali is 1.0 (0.2-20);

the solvent is organic solvent, water or their mixture;

the above organic solvent is C₁- C₆Halogenated hydrocarbon of (2), 1, 4-dioxane, C₁- C₆One or the mixture of alkyl alcohol, diethyl ether, acetonitrile, 2,2, 2-trimethyl acetonitrile, tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide, hexamethylphosphoramide, toluene and benzene;

the dehydrating agent is 3A, 4A, 5A molecular sieve or acid-washed 3A molecular sieve or one of anhydrous sodium sulfate, anhydrous calcium sulfate, anhydrous copper sulfate and anhydrous magnesium sulfate or a mixture thereof;

the above inorganic base means a hydroxide, hydride or carbonate of a monovalent metal; the organic base is potassium tert-butoxide, sodium tert-butoxide, tert-butyllithium, sodium methoxide, sodium ethoxide or magnesium methoxide;

the inorganic acid is HCl orH₂SO₄(ii) a The organic acid is acetic acid or formic acid;

the pseudospirostanol aglycone is pseudodiosgenin [. DELTA. ]⁵, Y = H, H, 25(R)]。

2. The method of claim 1, wherein the reducing agent is zinc/acetic acid solution or indium/acetic acid or indium/ammonium chloride aqueous solution.

3. The method of claim 1, wherein the lewis acid is one of trialkylsilyl triflate, boron trifluoride etherate, silver triflate, copper triflate, zinc triflate, scandium triflate, lanthanum triflate, ytterbium triflate, indium triflate, trifluoromethanesulfonate, perchloric acid, tetrafluoroboric acid, tetrakis (pentafluorophenyl) borate, or bis (trifluoromethanesulfonyl) imide.

4. The method of claim 1, wherein said acyl group is C₂- C₈Fatty acyl radicals or C₆- C₁₀An aromatic acyl group.

5. The method of claim 3, wherein said trialkylsilyl group is C₁- C₆Trialkyl silicon base.