CN102731610A - 26-thio or -seleno spirostanol saponin, synthesis method and application thereof - Google Patents

Abstract

The invention relates to 26-thio or -seleno spirostanol saponin and a chemical synthesis method of the 26-thio or -seleno spirostanol saponin. The method comprises the following steps: (1) 26-sulphonate of pseudospirostanol saponin is reacted with nucleophilic reagent of sulphur or selenium to obtain 26-S or -Se substitution product; (2) the substitution product is reacted with base or reducer and acid to obtain 26-thio or -seleno spirostanol sapogenin; (3) the obtained 26-thio or -seleno spirostanol sapogenin is reacted with acyl and silicon group protected rhamnose trichloroimidate donor to obtain potato trisaccharide thioglycoside; then, potato trisaccharide thioglycoside is hydrolyzed and reacted with tritox to be transformed to potato trisaccharide trichloroimidate donor; (4) potato trisaccharide trichloroimidate donor is reacted with acyl, alkyl and silicon group protected glycosyl donor, the fully-protected potato trisaccharide is introduced into 3-OH of the 26-thio or -seleno spirostanol sapogenin to obtain fully-protected 26-thio or seleno spirostanol saponin having alpha and beta configurations; (5) all protecting groups are removed to obtain 26-thio or -seleno spirostanol saponin. The compound has strong anti-tumour activity, and can be used for preparing anti-tumour drugs.

Description

26-position thio or selenospirosteroid 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, one of the most representative spirostanoid saponins, has become the most widely used medical saponin [Viviane, S. P.; Alemandre, T. C.; Taketa, G. G. 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 sulfur- or selenium-substituted spirostangenins [(a) Uhle, F. C.; J. Org. Chem. 1962 , 27 , 2797–2799; (b) Quan, H. J.; Koyanagi. J. ; Ohmori, K.; Uesato, S.; Tsuchido, T.; Saito, S. Eur. J. Med. Chem. 2002 , 37 , 659–669], but no information about the 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 are 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 selenogenin:

Under the condition of 0-150℃, the 26-position S or Se substituted pseudospirogenin reacts with alkali, or reducing agent and acid to obtain 26-position thio or selenogenin;

(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-glucosinolate and acyl- and 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) The 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 selenospiroside:

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 the 26-position pseudospirogenin reacts with a nucleophile of sulfur or selenium at 0-150°C for 1-10 hours to obtain a 26-position S or Se substituted Product; Wherein, the mol ratio of sulfonate 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 ;

Described 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 and at 0-150°C, the above-mentioned S or Se substituted pseudospironogenin 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 alkali 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 0.1-1 mol/L methanolic or ethanol solution of hydrochloric acid.

(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-oxy-benzoyl-β-D-glucoglucoside and acyl, Silicon-protected rhamnose trichloroimidate donor was reacted for 1-10 hours to obtain potato triose glucosinolate, which was then converted into trisaccharide trichloroimidate donor by hydrolysis and reaction with trichloroacetonitrile;

3, The molar ratio of 6-di-oxo-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 di-oxygen-benzoyl-β-D-glucosinolate to 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 spirogenin and acyl- or / and silicon-protected potato trichloride Imino ester donor reaction for 1-10 hours, introducing a 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 selenospirogenin to the glycosyl acceptor and accelerator is 1.0:(1.0-5.0):(0.05-0.5); The weight ratio of yuan to 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 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 the solvent, under the condition of -20 - 60 ℃, the fully protected α and/or β configurations of 26-position thio or selenospirosteroid saponins are reacted for 10 - 60 hours in the presence of alkali 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

Figure 1 is the synthesis process of 26-position thio or selenodiosgenin.

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

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 argon protection, sulfonate 1 (369 mg, 0.65 mmol) [(a) Uhle, F. C. 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 26-position thioacetyl substituted Compound 2 (295 mg, 0.62 mmol, 96%).

= -22.8 (c 1.00, CHCl₃)； [α]

= - 22.8 ( c 1.00, CHCl ₃ );

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

¹³ C NMR (150 MHz, 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, 32.6, 3.3 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 _C29H45O3S : 473.3084 _; Found: _{C29H45O3S 473.3090} .

Synthesis of compound 3 :

Under the protection of argon, 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) 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, filtered Collect solids. After the solid was dried, it was dissolved in 20 mL of ethanol/water (EtOH: H ₂ O = 19: 1) mixed solution under the protection of argon, and then 0.5 mL of 37% concentrated HCl was added, heated to reflux for 5 h, and then cooled , poured into ice water, filtered, collected the solid, and further purified by flash column chromatography to obtain 26-position S-substituted spironogenin 3 (209 mg, 0.48 mmol, 87%).

= -162.7 (c 1.10, CHCl₃)； [α]

= - 162.7 ( c 1.10, CHCl ₃ );

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

¹³ C NMR (150 MHz, 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, 312.0, 32.0, 31.6, 31.41, 31.38, 22.4, 20.8, 19.4, 16.5, 16.2.

_{HRMS (} ESI): [M + H] ⁺ _, Calcd. for _{C27H43O2S :} 431.2978; Found: C27H43O2S _431.2983 .

Synthesis of compound 4 :

Under argon protection, CsOH·H ₂ O (76 mg, 0.45 mmol), selenium powder (23.7 mg, 0.30 mmol) were added to 2 mL DMF, and N ₂ H ₄ ·H ₂ O (55 μl , 0.90 mmol), after stirring for 2 h, sulfonate 1 (171 mg, 0.30 mmol) was added, then the temperature was raised to 60°C, stirring was continued for 4 h, 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%).

= 34.2(c 1.21, CHCl₃)； [α]

= 34.2( c 1.21, CHCl ₃ );

¹ H NMR (600 MHz, CDCl ₃ ) δ 5.34 (d, 2H, J = 4.8 Hz), 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, 7.8 Hz), 2.45 (d, 2H, J = 10.2 Hz), 2.30 (m, 4H), 2.22 (t, 4H, J = 10.8 Hz) 1.58 (s, 6H), 1.01 (s, 6H), 0.99 (d, 6H, J = 6.6 Hz), 0.68 (s, 6H).

¹³ C NMR (150 MHz, 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, 3.2, 33. , 31.2, 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, and then added to 20 mL of 10% KOH in ethanol/water mixed solution (EtOH: H ₂ O = 1: 1), at room temperature After stirring 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%).

= -192.8 (c 1.00, CHCl₃)； [α]

= - 192.8 ( c 1.00, CHCl ₃ );

¹ H NMR (600 MHz, CDCl ₃ ): δ 5.34 (d, 1H, J = 4.8 Hz), 4.64 (dd, 1H, J = 15.6, 7.8 Hz), 3.53-3.50 (m, 1H), 2.58 (t , 1H, J = 12.0 Hz), 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 = 7.2 Hz), 1.01 (s, 3H), 0.96 (d, 3H, J = 6.6 Hz), 0.80 (s, 3H).

¹³ C NMR (150 MHz, 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, 31.0, 32.6, 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% for 11α ; 27% for 11β ;,32% for 12α ; 46% for 12β ; (e) CH ₃ ONa, CH ₃ OH : CH ₂ Cl ₂ = 1 : 1, 99% for 13α ; 95% for 13β ; 90% for 14α ; 78% for 14β .

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. ] (1.73 g, 3.50 mmol), freshly activated 4? molecular sieve (2.30 g) was added to anhydrous CH ₂ Cl ₂ (30 mL),- After stirring at 30°C 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, N. I.; Ohashi, K.; Sakagami, M.; Yoshikawa, M.; Shibuya, Chem. Pharm. Bull. 1989, 37 , 1131–1133; (b) Cheng, M. S.; Wang, Q. L.; Tian, Q.; Song, H. Y.; Liu, Y. X.; X.; Miao, H. D.; Yao, X. S.; Yang, Z. J. Org. Chem. 2003, 68 , 3658–3662.] (4.56 g, 10.49 mmol) in CH ₂ Cl ₂ (5 ml), -30°C After the reaction was stirred for 2 h, TLC detected that 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 reaction was dissolved in 30 mL of acetone/water (9: 1), and NBS (1.16 g, 6.50 mmol) was added under ice-cooling. After stirring for 30 min, TLC showed that the reaction was complete. Add saturated NaHCO ₃ dropwise to quench the reaction, concentrate under reduced pressure to remove acetone, and dilute the residue with CH ₂ Cl ₂ , separate and collect the organic phase, then wash with saturated NaHCO ₃ and saturated NaCl successively, dry over anhydrous NaSO ₄ , and filter , concentration, and flash column chromatography to obtain compound 9 (2.34 g, 2.51 mmol, two-step total yield 72%).

Under argon protection, compound 9 (1.92 g, 2.06 mmol) was dissolved in CH ₂ Cl ₂ (25 mL), DBU (125 μL, 0.82 mmol) was added under ice cooling, and after stirring for 5 min, Cl ₃ CCN was slowly added dropwise (2.06 mL, 20.58 mmol) was reacted at room temperature for 2 h, 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 (2.02 g, 1.87 mmol, 91%).

= 103.2 (c 1.05, CHCl₃)； [α]

= 103.2 ( c 1.05, CHCl ₃ );

¹ H NMR (600 MHz, 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 = 3.6 Hz), 5.89 (t, 1H, J = 10.2 Hz), 5.20 (dd, 1H, J = 9.6, 3.0 Hz), 5.15 (dd, 1H, J = 3.6, 1.8 Hz), 5.11 (dd, 1H, J = 9.6, 3.6 Hz), 4.93-4.87 (m, 3H), 4.83-4.80 (m, 2H), 4.79 (d, 1H, J = 1.2 Hz), 4.51 (dd, 1H, J = 12.6, 3.6 Hz), 4.33-4.30 (m, 1H), 4.10 (t, 1H, J = 9.6 Hz), 4.05 (dd, 1H, J = 10.2, 4.2 Hz), 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 = 6.6 Hz), 0.69 (d, 3H, J = 6.0 Hz).

¹³ C NMR (150 MHz, 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, 99.4, 99.1, 94.1, 90.6, 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, 16.18, 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 ? Trifluoromethylsilyl trifluoromethanesulfonate (TMSOTf ) (7 μL, 0.04 mmol, 0.2 equiv.), after stirring for 10 min, the ester-protected trisaccharide donor 10 (280 mg, 0.26 mmol, 1.30 equiv.), after stirring at -30°C for 4 h, 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%).

Compound 11α :

= 76.6 (c 0.80, CHCl₃)； [α]

= 76.6 ( c 0.80, CHCl ₃ );

¹ H NMR (600 MHz, 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 = 9.6 Hz) , 5.25 (d, 1H, J = 4.2 Hz ), 5.22 (dd, 1H, J = 10.2, 3.6 Hz), 5.17 (dd, 1H, J = 10.2, 3.6 Hz), 5.15-5.11 (m, 1H), 5.05 (d, 1H, J = 3.6 Hz), 4.90 (t, 1H, J = 9.6 Hz), 4.89 (t, 1H, J = 10.2 Hz), 4.85 (d, 1H, J = 1.8 Hz), 4.77- 4.72 (m, 3H), 4.63 (dd, 1H, J = 15.6, 7.2 Hz), 4.52 (dd, 1H, J = 12.6, 5.4 Hz), 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, 3.0 Hz), 3.48-3.42 (m, 1H), 2.53 (t, 1H, J = 13.2 Hz), 2.47 (d , 2H, J = 7.8 Hz), 2.28 (d, 1H, J = 12.6 Hz), 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 = 6.0 Hz), 1.05 (s, 3H), 1.00 (d, 3H, J = 7.2 Hz), 0.92 (d, 3H, J = 6.6 Hz), 0.80 (s, 3H), 0.69 (d, 3H, J = 6.0 Hz).

¹³ C NMR (150 MHz, 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, 128.2, 121.9, 99.4, 99.0, 97.5, 96.2, 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 _C71H92O23SNa : 1367.5642; Found _: 1367.5691.

Compound 11β :

= 34.6 (c 0.80, CHCl₃)； [α]

= 34.6 ( c 0.80, CHCl ₃ );

¹ H NMR (600 MHz, 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 = 9.4 Hz) , 5.33 (d, 1H, J = 5.4 Hz), 5.14-5.11 (m, 2H), 5.09 (m, 1H), 4.95-4.93 (m, 1H), 4.89 (t, 1H, J = 10.4 Hz), 4.85 (t, 1H, J = 10.2 Hz), 4.83 (brs, 1H), 4.77 (d, 1H, J = 11.52 Hz), 4.74 (brs, 1H), 4.65 (d, 1H, J = 7.68 Hz), 4.64-4.60 (m, 1H), 4.49 (dd, 1H, J = 12.0, 5.4 Hz), 4.35-4.30 (m, 1H), 3.94 (t, 1H, J = 9.6 Hz), 3.85-3.81 (m, 1H), 3.77 (t, 1H, J = 7.2 Hz), 3.70-3.66 (m, 1H), 3.58-3.53 (m, 1H), 2.52 (t, 1H, J = 11.4 Hz), 2.38 (d, 1H , J = 10.8 Hz), 2.28 (d, 1H, J = 11.4 Hz), 2.23 (t, 1H, J = 12.6 Hz), 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 = 6.0 Hz), 1.00 (d, 3H, J = 6.6 Hz), 0.93 (s, 3H), 0.90 (d , 3H, J = 6.0 Hz), 0.78 (s, 3H), 0.66 (d, 3H, J = 6.0 Hz).

¹³ C NMR (150 MHz, 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, 129.0, 128.8, 128.34, 128.31, 121.9, 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, 22.73, 20.68, 20.62, 20.2, 19.1, 16.8, 16.5, 16.1.

_HRMS (ESI): [M + Na] ⁺ , Calcd. for _C71H92O23SNa : _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 ? Trifluoromethylsilyl trifluoromethanesulfonate (TMSOTf ) (6 μL, 0.034 mmol, 0.2 equiv.), after stirring for 10 min, the ester-protected trisaccharide donor 10 (238 mg, 0.22 mmol, 1.30 equiv.), stirred at -78°C for 1 h, and continued stirring at -40°C for 3 h, TLC showed that the reaction was complete. After the reaction was quenched with triethylamine, 12α (76 mg, 0.055 mmol, 32%) and 12β (110 mg, 0.079 mmol, 46%) were obtained as white solids after filtration, concentration, and flash column chromatography.

Compound 12α :

= -1.9 (c 1.40, CHCl₃)； [α]

= - 1.9 ( c 1.40, CHCl ₃ );

¹ H NMR (600 MHz, CDCl ₃ ): δ 8.06 (d, 2H, J = 4.2 Hz), 8.04 (d, 2H, J = 4.2 Hz), 7.57-7.52 (m, 2H), 7.44-7.40 (dd , 4H, J = 13.8, 7.8 Hz), 5.78 (t, 1H, J = 9.6 Hz), 5.26 (d, 1H, J = 4.2 Hz), 5.23 (dd, 1H, J = 10.2, 3.0 Hz), 5.18 (dd, 1H, J = 9.6, 3.6 Hz), 5.15-5.12 (m, 1H), 5.06 (d, 1H, J = 3.6 Hz), 4.93-4.88 (m, 2H), 4.86 (brs, 1H), 4.78-4.76 (m, 1H), 4.74 (d, 1H, J = 13.2 Hz), 4.66 (dd, 1H, J = 15.6, 7.2 Hz), 4.53 (dd, 1H, J = 12.0, 4.8 Hz), 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, 3.6 Hz), 3.48-3.43 (m, 1H), 2.59 (t, 1H, J = 12.0 Hz), 2.48 (d, 2H, J = 7.8 Hz), 2.37 (d, 1H, J = 11.4 Hz), 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 = 6.0 Hz), 1.06 ( s, 3H), 1.02 (d, 3H, J = 7.2 Hz), 0.96 (d, 3H, J = 6.6 Hz), 0.81 (s, 3H), 0.69 (d, 3H, J = 6.0 Hz).

¹³ C NMR (150 MHz, 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, 121.9, 99.4, 99.0, 97.9, 96.2, 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 : 1415.5087 _{for C71H92O23SeNa} ; Found: 1415.5119.

Compound 12β :

= -44.5 (c 1.70, CHCl₃)； [α]

= - 44.5 ( c 1.70, CHCl ₃ );

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

¹³ C NMR (150 MHz, CDCL ₃ ): Δ 169.93, 169.87, 169.80, 169.5, 168.8, 165.7, 164.9, 140.0, 132.9, 130.0, 129.7, 129.34, 128.29, 99.4, 98.9, 98.9, 98.9, 98.9, 98.9, 98.9, 98.9, 98.9, 98.9, 98.9, 98.9, 98.9, 98.9, 98.9, 98.9, 99.4, 98.9, 99.9, 99.4, 98.9, 99.4, 98.9, 99.4, 98.9, 99.4, 98.9, 99.4. 98.0, 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 _{C71H92O23SeNa} : 1415.5087; found: 1415.5144.

Synthesis of Compound 13α :

Under argon protection, compound 11α (40 mg, 0.0297 mmol) was dissolved in a mixed solvent of 2 ml methanol/dichloromethane (MeOH: CH ₂ Cl ₂ = 1: 1), and then sodium methoxide (24 mg, 0.44 mmol, 14.6 equiv.) in methanol solution (230 μL), stirred at room temperature for 48 h, 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 (26 mg, 0.0294 mmol, 99%) as a white solid.

= -46.0 (c 0.09, CHCl₃ : 含有8%水的MeOH  = 1 : 1)； [α]

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

¹ H NMR (600 MHz, pyridine- d ₅ ): δ 5.89 (s, 1H), 5.81 (s, 1H), 5.46 (d, 1H, J = 3.0 Hz), 5.27 (d, 1H, J = 3.6 Hz ), 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 = 10.2 Hz), 4.21 (d, 1H, J = 12.0 Hz), 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 = 10.8 Hz), 1.66 (d, 3H, J = 6.0 Hz), 1.64 (d, 3H, J = 6.0 Hz), 1.07 (d, 3H, J = 6.0 Hz), 0.82 (m, 6H), 0.78 (s, 3H).

¹³ C NMR (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, 6, 61.2, 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, 18.3, 18.4.

HRMS (ESI): [ _M + Na] ⁺ , Calcd. for _C45H72O15SNa , _907.4484 ; Found: 907.4505.

Synthesis of Compound 13β :

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

= -97.4 (c 0.85, CHCl₃ : 含有8%水的MeOH  = 1 : 1)； [α]

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

¹ H NMR (600 MHz, 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 = 12.6 Hz), 3.86-3.80 (m, 1H), 3.60 (d, 1H, J = 7.8 Hz), 2.74 (d, 1H, J = 12.0 Hz), 2.68 (t, 1H, J = 12.0 Hz), 2.60 (t, 1H, J = 12.6 Hz), 2.30 (d, 1H, J = 12.0 Hz), 1.72 (d, 3H, J = 6.0 Hz), 1.58 (d, 3H, J = 6.0 Hz) , 1.07 (d, 3H, J = 7.2 Hz), 1.01 (s, 3H), 0.79 (m, 6H).

¹³ C NMR (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, 72.2, 6 , 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, 22.2, 20.8, 18.19

HRMS (ESI): [ _M + Na] ⁺ , Calcd. for _C45H72O15SNa , _907.4484 ; Found: 907.4501.

Synthesis of Compound 14α :

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

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

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

¹³ C NMR (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, 30.2.0, 7 , 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, 17.3.

HRMS (ESI): [M + Na] ⁺ , Calcd. for C ₄₅ H ₇₂ O ₁₅ SeNa, 955.3929; Found: 955.3950.

Synthesis of Compound 14β :

According to the same method as compound 13α , compound 12β (40 mg, 0.0287 mmol) was deprotected to obtain white solid 14β (21 mg, 0.0225 mmol, 78%).

= -148.7 (c 0.80, CHCl₃ : 含有8%水的MeOH = 1 : 1)； [α]

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

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

¹³ C NMR (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, 72.2, 9, 7 63.2, 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, 16.4, 18.4.

HRMS ( _ESI ): [M + H] ⁺ , Calcd. for _C45H73O15Se , _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, H. C.; Yu, B. Tetrahedron Lett. 2001 , 42 , 2405–2407] inhibitory activity on 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 (10)

1. The 26-position thio or selenospirosterol saponin is characterized in that its general structural formula is as follows:

in, 5 and 6 are single or double bonds (Δ ⁵ ); X = S or Se; Y = H and H, O or OH and H; R ¹ - R ⁸ are any one of hydrogen, acyl or alkyl; Potatotriose and its derivatives are connected with aglycon by α or β glycosidic bonds; The absolute configuration of the 22-position carbon is R , and the absolute configuration of the 25-position carbon is R or S. 2. The spirosteroid saponin according to claim 1, characterized in that the acyl group is a C ₂ -C ₆ linear or branched aliphatic acyl group or a C ₆ -C ₁₀ aromatic acyl group. 3. The spirosteroid saponin as claimed in claim 1, characterized in that said alkyl group is methyl, ethyl, n-propyl or isopropyl. 4. the synthetic method of 26 thios as claimed in claim 1 or selenospirulina saponin, it is characterized in that the method comprises the steps: (1) Synthesis of 26-position thio- or seleno-pseudospironin: In a solvent or in the presence of a phase transfer catalyst, the 26-position p-toluenesulfonate of pseudospirogenin reacts with a sulfur or selenium nucleophile at 0-150°C for 1-10 hours to obtain the corresponding sulfur or selenium Substitution product; wherein, the molar ratio of tosylate and nucleophile is 1:(1.0 - 5.0); the nucleophile is KSCOCH ₃ , Na ₂ S ₂ , Li ₂ S ₂ , Cs ₂ S ₂ , K ₂ S ₂ or one of KSeCOCH ₃ , Na ₂ Se ₂ , Li ₂ Se ₂ , Cs ₂ Se ₂ , K ₂ Se ₂ ; the phase transfer catalyst is tetrabutylammonium bromide , one of tetrabutylammonium iodide, tetrabutylammonium bisulfate, polyethylene glycol-400; (2) Preparation of 26-position thio or selenogenin: In the solvent and at 0-150℃, the 26-position thio- or seleno-psirosteroid saponin reacts with a base or a reducing agent, and then reacts with an acid for 0.1-10 hours to obtain the 26-position thio- or seleno-spirulina Saponin; the mol ratio of 26-position thio or seleno-pseudospirosteroid saponin to base or reducing agent is 1.0:(2.0-20.0):(2.0-5.0); the base is an organic base or an inorganic base; acid is an organic or inorganic acid; (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 was reacted for 1-10 hours to obtain potato trisaccharide glucosinolate, which was then converted into trisaccharide trichloroimidate donor through hydrolysis and reaction with trichloroacetonitrile; 3, The molar ratio of 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-di- The weight ratio of oxygen-benzoyl-β-D-glucosinolate to dehydrating agent is 1.0:(3.0 - 10.0); (4) The introduction of the 3-position sugar group: In the presence of an organic solvent and a dehydrating agent, at -78 - 40°C, with a Lewis acid or a protonic acid as a promoter, the 26-position thio or selenospironin and the acyl or silicon-protected glycosyl donor React for 1 - 10 hours, introduce fully protected potato triose in its 3 OH position, and obtain fully protected spirostanoid saponins in α or/and β configurations; among them, the 26-position thio or selenospironin The molar ratio of glycosyl donor and accelerator is 1.0:(1.0 - 5.0):(0.05 - 0.5); the weight ratio of 26-position thio or selenogenin to dehydrating agent is 1.0:(3.0 - 10.0) ; (5) Synthesis of 26-position thio or selenospiroside: In a solvent at - 20 - 60°C, fully protected spirostanoid saponins in the α or/and β configurations are reacted with the base for 10 - 60 hours respectively, and all protective groups are removed to obtain the α or/and β configurations The 26-position thio or selenospirulina saponin; the base is an inorganic or organic base; the molar ratio of the fully protected spirostanol to base is 1.0:(0.2-20); Above-mentioned solvent is organic solvent, water or their mixture; The above-mentioned organic solvents are C ₁ - C ₆ halogenated hydrocarbons, 1, 4-dioxane, C ₁ - C ₆ alkyl alcohols, diethyl ether, acetonitrile, 2, 2, 2-trimethylacetonitrile, tetrahydrofuran , N, N-dimethylformamide, N, N-dimethylacetamide, hexamethylphosphoramide, toluene, benzene or a mixture thereof; The dehydrating agent is one of 3?, 4?, 5? molecular sieve or pickled 3? molecular sieve or anhydrous sodium sulfate, anhydrous calcium sulfate, anhydrous copper sulfate, anhydrous magnesium sulfate or their mixture ; The above-mentioned inorganic bases refer to hydroxides, hydrides or carbonates of monovalent metals; the organic bases are potassium tert-butoxide, sodium tert-butoxide, tert-butyllithium, sodium methylate, sodium ethylate or magnesium methoxide; The aforementioned inorganic acid is HCl or H ₂ SO ₄ ; the organic acid is acetic acid or formic acid. 5. synthetic method as claimed in claim 4, is characterized in that described pseudospirogenin is pseudodiosgenin [△ ⁵ , Y=H, H, 25 ( R )]; Pseudoyam saponin [ △ ⁵ , Y = H, H, 25( S )]; Pseudotacogenin [5α-H, Y = H, H, 25( R )]; Pseudotacogenin [5α-H, Y = H, H, 25( S )]; Pseudohecogenin [5α-H, Y = O, 25( R )]; Pseudohecogenin [5α-H, Y = O, 25( S )]; Pseudosmilagenin [5β-H, Y = H, H, 25( R )]; Pseudotimogenin [5β H, Y = H, H, 25( S )]; Pseudoportogenin [△ ⁵ , Y = O, 25( R )]; pseudoneopergenin [△ ⁵ , Y = O, 25( S )]; pseudoisoziapaggenin [△ ⁵ , Y = β – OH, α -H, 25( S )]. 6. synthetic method as claimed in claim 4, is characterized in that described reducing agent is zinc/acetic acid solution or indium/acetic acid or indium/ammonium chloride aqueous solution. 7. the synthetic method as claimed in claim 4 is characterized in that described Lewis acid is trialkylsilyl trifluoromethanesulfonate, boron trifluoride ether, silver trifluoromethanesulfonate, trifluoromethanesulfonate copper triflate, zinc triflate, scandium triflate, lanthanum triflate, ytterbium triflate, indium triflate, triflate, perchloric acid, tetrafluoromethanesulfonate One of boric acid, tetrakis(pentafluorophenyl)boronic acid or bis(trifluoromethanesulfonyl)imide. 8. The synthesis method according to claim 4, characterized in that the acyl group is a C ₂ -C ₈ fatty acyl group or a C ₆ -C ₁₀ aromatic acyl group. 9. The synthesis method according to claim 4, characterized in that the trialkylsilyl group is a C ₁ -C ₆ trialkylsilyl group. 10. The use of the 26-position thio or selenospiroside as claimed in claim 1 in the preparation of antitumor drugs.

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