A novel anti-cancer drug TNBG-glycoside conjugate and preparation and application thereof

Abstract

The application provides a novel anticancer drug TNBG-sugar conjugate and a preparation and application thereof, and belongs to the field of chemical medicines.The conjugate is a compound shown in formula I and formula IV or a salt or a stereoisomer thereof.The anticancer drug TNBG-sugar conjugate and a preparation prepared by using suitable excipients or auxiliary ingredients have a good inhibitory effect on cancer cells such as lung cancer and liver cancer, have low or basically no toxicity to normal cells, can be used for the prevention and treatment of lung cancer, liver cancer and other cancers, and have a good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of chemical medicine and relates to a novel anticancer drug TNBG-glycoconjugate and its preparation and application. Background Technology

[0002] Sugars, also known as carbohydrates, possess advantages such as low toxicity and good biocompatibility, playing a crucial role in biological processes including cell recognition, microbial infection, immune response, and cell proliferation. As a novel therapeutic approach, drug-glycoconjugates have demonstrated high efficacy and specificity. Currently, over 170 drug-glycoconjugates have been successfully approved for marketing by the Food and Drug Administration (FDA) in countries including the United States, including sugars (oligosaccharides / polysaccharides), small molecule glycosides and glycomimetic compounds, glycopeptides and glycoproteins, and sugar-based vaccines. Existing research shows that in anti-tumor therapy, anticancer drug-glycoconjugates can exert their anticancer effects, similar to the binding of drugs to antibodies. This binding mechanism, through sugar modification, can increase drug solubility, in vivo stability, prolong the duration of drug action, and improve therapeutic efficacy. Furthermore, sugars can be used as targeting molecules, recognizing and binding to tumor cells through specific receptors or glycan structures on the cell surface, thereby achieving therapeutic goals. This not only possesses high specificity but also reduces damage to normal cells, improving the safety and effectiveness of treatment. In conclusion, as drug-glycoconjugates become increasingly widely used in the pharmaceutical field, we firmly believe that they will make a greater contribution to human health in the future.

[0003] Tetrazanbigen (TNBG) is a novel azasteroid compound with excellent in vitro and in vivo antitumor activity and no cross-resistance with other commonly used antitumor drugs. Experimental studies have shown that TNBG activates the transcription factor SREBP1 (Sterol-regulated element-binding protein transfer protein) and peroxisome proliferator-activated receptor γ (PPARγ), inhibiting the lipid transport MTTP (Microsomal triglyceride transfer protein), leading to lipid accumulation in tumor cells and ultimately cell death due to lipotoxicity. Therefore, TNBG is a novel non-cytotoxic antitumor drug that specifically induces lipid accumulation in tumor cells through multiple targets, resulting in lipotoxicity. However, TNBG's high lipid solubility and poor water solubility affect absorption, blood concentration, and tissue concentration, severely limiting its drug development potential.

[0004] In summary, the strategy of conjugating the anticancer drug TNBG with glycosides can improve its water solubility and enhance its antitumor activity. Therefore, the design and development of novel anticancer drug TNBG-glycoside conjugates will undoubtedly play a very important role and have significant implications. Summary of the Invention

[0005] This invention is based on the anticancer drug TNBG, and through structural modification, innovatively obtained a series of novel anticancer drug TNBG-glycoconjugates with good drug-like properties.

[0006] One object of the present invention is to provide a method for preparing and using the novel anticancer drug TNBG-glycoconjugates shown in Formulas I and IV.

[0007] Another object of the present invention is to provide a method for making compounds of formula I and IV, or salts thereof, or stereoisomers thereof.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] In a first aspect, the present invention provides an anticancer drug TNBG-glycoconjugate of Formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0010]

[0011] in,

[0012] n is an integer selected from 1 to 6;

[0013] G is selected from monosaccharides (glucose, galactose, mannose, etc.), polysaccharides (glucan, lactose, etc.), etc.

[0014] R1-R8 are selected from hydrogen, halogen, C1-C8 alkyl, C1-C8 alkoxy, hydroxyl, nitro, amino, carboxyl, formula II or formula III, etc.

[0015]

[0016] n1 is an integer selected from 1 to 8, with the sign... Represents a single bond or a double bond;

[0017] R9 and R 10 Selected from hydrogen, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, cyclopropyl or cyclobutyl; and methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, cyclopropyl or cyclobutyl with substituents, etc.

[0018] n2 is an integer selected from 1 to 8, with the sign... Represents a single bond or a double bond;

[0019] n3 is an integer selected from 1 to 2, with the sign... Represents a single bond or a double bond;

[0020] X is selected from oxygen, nitrogen, or carbon, etc.;

[0021] R 11 It is selected from hydrogen, halogen, C1-C8 alkyl, C1-C8 alkoxy, hydroxyl, nitro, amino or carboxyl, etc.

[0022] In some preferred embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein n is usually selected from an integer 3, and G may be selected from D-glucose, D-galactose, etc.

[0023] This invention provides a synthetic route for the anticancer drug TNBG-glycoconjugate shown in Formula I, as follows:

[0024]

[0025] The synthesis of the anticancer drug TNBG-glycoconjugate shown in Formula I has the following characteristics:

[0026] First, the lead compound of the anticancer drug TNBG, namely key intermediate 10, was synthesized by referring to the mature route of the research group. Then, the target compound of formula I was obtained through substitution reaction, glycosylation reaction and deacetylation reaction.

[0027] The substitution reaction is the substitution of hydrogen on a secondary amine with 1-bromoethanol or propanol under strong alkaline conditions (NaH (60%)); the glycosylation reaction is the reaction of a hydroxyl group with a chiral fixed tetraacetyl bromide under the catalysis of a heavy metal salt; the deacetylation reaction is carried out in a methanol solution in an alkaline (potassium carbonate) environment to obtain target compounds I-1 and I-2.

[0028] In a second aspect, the present invention provides an anticancer drug TNBG-glycoconjugate of Formula IV, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0029]

[0030] in,

[0031] n is an integer selected from 1 to 6;

[0032] G is selected from monosaccharides (glucose, galactose, mannose, etc.), polysaccharides (glucan, lactose, etc.), etc.

[0033] R1-R4 are selected from hydrogen, halogen, C1-C8 alkyl, C1-C8 alkoxy, hydroxyl, nitro, amino, carboxyl, formula V or formula VI, etc.

[0034]

[0035] n1 is an integer selected from 1 to 8, with the sign... Represents a single bond or a double bond;

[0036] R5 and R6 are selected from hydrogen, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, cyclopropyl, or cyclobutyl; and methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, cyclopropyl, or cyclobutyl with substituents, etc.

[0037] n2 is an integer selected from 1 to 8, with the sign... Represents a single bond or a double bond;

[0038] n3 is an integer selected from 1 to 2, with the sign... Represents a single bond or a double bond;

[0039] X is selected from oxygen, nitrogen, or carbon, etc.;

[0040] R7 is selected from hydrogen, halogen, C1-C8 alkyl, C1-C8 alkoxy, hydroxyl, nitro, amino or carboxyl, etc.

[0041] In some preferred embodiments, the compound of formula IV or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein n is usually selected from integers 2 or 3; R 1-4 The hydrogen and alkyl groups are preferably methyl groups, the halogens are preferably F and Cl, the alkoxy groups are preferably methoxy groups, etc., and G is preferably D-glucose, D-galactose, etc.

[0042] This invention provides a synthetic route for the anticancer drug TNBG-glycoconjugate shown in Formula IV, as follows:

[0043]

[0044] The synthesis of the anticancer drug TNBG-glycoconjugate shown in Formula IV has the following characteristics:

[0045] The synthesis proceeded through cyclization, chlorination, and cyclization to obtain key intermediate 14, which was then subjected to substitution, glycosylation, and deacetylation to obtain the target compound of formula IV.

[0046] The cyclization reaction is the reaction of the starting material with diethyl oxalate under acidic conditions (dilute hydrochloric acid); the chlorination reaction is the conversion of the hydroxyl group into a chlorine atom using phosphorus oxychloride; the cyclization reaction is the formation of a four-membered ring key intermediate 14 with 2-aminomethylpiperidine under alkaline conditions (potassium carbonate); the substitution reaction is the replacement of the hydrogen atom on the secondary amine with 1-bromoethanol or propanol under strongly alkaline conditions (NaH (60%)); the glycosylation reaction is the reaction of the hydroxyl group with a chiral fixed tetraacetyl bromide under the catalysis of a heavy metal salt; the deacetylation reaction is carried out in a methanol solution in an alkaline (potassium carbonate) environment to obtain the target compounds IV-1 to IV-24.

[0047] Thirdly, the present invention provides the following representative compounds or their pharmaceutically acceptable salts or stereoisomers:

[0048]

[0049]

[0050] Fourthly, the present invention provides specific uses of the compounds of Formula I and Formula IV, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, in the preparation of antitumor drugs;

[0051] The prepared antitumor drug is a drug for treating cancer, characterized in that: preferably, the cancer is lung cancer, liver cancer, etc.;

[0052] A drug is a formulation prepared by using the compounds of Formula I and Formula IV, or their pharmaceutically acceptable salts or stereoisomers as active ingredients, combined with suitable excipients or auxiliary ingredients. Attached Figure Description

[0053] Figure 1 This is a schematic diagram of the general structural formula of this invention patent. Detailed Implementation

[0054] Unless otherwise stated, all reagents and instruments used in this invention are commercially available, and all chemical reagents are AR grade, requiring no further purification. The invention will now be further described in detail with reference to the embodiments, but is not limited to these embodiments.

[0055] The structures of the compounds in the following examples were determined primarily by nuclear magnetic resonance (NMR) or high-resolution mass spectrometry (HRMS). NMR measurements were performed using a Bruker AV-600 NMR spectrometer (USA) with deuterated chloroform (CDCl3) or deuterated dimethyl sulfoxide (DMSO-d6) as solvent and tetramethylsilane (TMS) as internal standard; HRMS measurements were performed using a Waters G2-S QTOF-MS mass spectrometer (USA); melting point data were collected using a WRS-2C melting point apparatus (Shanghai, China).

[0056] Firstly, the synthesis of compounds of formula I:

[0057] Example 1: Synthesis of key intermediate 6

[0058] The synthetic route for key intermediate 6 is as follows:

[0059]

[0060] Reagent and conditions a are: chloroacetyl chloride, sodium carbonate, dichloromethane, 0℃;

[0061] The reagents and conditions b are: potassium phthalimide, N,N-dimethylformamide, and 110℃;

[0062] The reagents and conditions for c are: phosphorus pentoxide, zinc chloride, acetonitrile, and 120℃;

[0063] The reagents and conditions d are: sodium triacetoxyborohydride, methanol, and dichloromethane;

[0064] The reagents and conditions e are: hydrazine hydrate, methanol, dichloromethane, room temperature, and concentrated hydrochloric acid;

[0065] The specific preparation method is as follows:

[0066] (1) Preparation of 2-chloro-N-phenylethylacetamide (compound 2)

[0067] Take a clean three-necked flask, add 60 mL of dichloromethane and anhydrous sodium carbonate (14.4 g, 0.135 mol, 1.5 eq), and add chloroacetyl chloride (12.3 mL, 0.154 mol, 1.7 eq) under stirring in an ice-salt bath. Slowly add a dichloromethane solution of β-phenylethylamine (11 g, 0.090 mol, leq), keeping the temperature below 0°C. Monitor the reaction by TLC until complete. Slowly pour the reaction solution into water, separate the aqueous layer, and extract successively with dichloromethane (2 x 60 mL) and saturated brine (20 mL). Combine the organic layers, dry with anhydrous sodium sulfate, concentrate, and bake dry to obtain a pale yellow solid (compound 2), 16.87 g, yield 94%.

[0068] (2) Preparation of 2-(1,3-dioxoisoindol-2-yl)-N-phenylacetamide (compound 3)

[0069] Compound 2 (16.87 g, 0.085 mol, 1 eq) was dissolved in 70 mL of N,N-dimethylformamide, and potassium phthalimide (20.55 g, 0.111 mol, 1.3 eq) was added with stirring. The mixture was heated to 110 °C and reacted for 5 h. The reaction solution was poured into cold water, and a large amount of white solid precipitated. The solid was filtered and dried to obtain a white solid (compound 3), 25.08 g, with a yield of 95.3%.

[0070] (3) Preparation of 2-((3,4-dihydroisoquinolin-1-yl)methyl)isoindole-1,3-dione (compound 4)

[0071] Take a clean eggplant-shaped flask and add compound 3 (25.08 g, 0.081 mol, 1 eq), phosphorus pentoxide (8.08 g, 0.057 mol, 0.7 eq), zinc chloride (3.88 g, 0.028 mol, 0.35 eq), phosphorus oxychloride (24.94 g, 0.163 mol, 2 eq) and 350 mL of acetonitrile in sequence. Heat to 90 °C with stirring and monitor the reaction until complete by TLC. Concentrate under reduced pressure and pour the reaction solution into ice water. Adjust the pH to weakly alkaline with alkaline water to precipitate a yellow solid. Filter and dry to obtain a yellow solid (compound 4), 23.08 g, yield 97.8%.

[0072] (4) Preparation of 2-((1,2,3,4-tetrahydroisoquinolin-1-yl)methyl)isoindole-1,3-dione (compound 5)

[0073] In a clean, round-bottomed flask, compound 4 (23.08 g, 0.079 mol, leq), 40 mL of methanol, and 100 mL of dichloromethane were added. The mixture was stirred at room temperature, and sodium triacetoxyborohydride (67.4 g, 0.318 mol, 4 eq) was added in four portions. The reaction was monitored by TLC until complete. The reaction solution was poured into water, and the pH was adjusted to alkaline with sodium carbonate. A partial solid was observed. The mixture was filtered, and the filter cake was washed with dichloromethane. The aqueous layer was extracted with dichloromethane (3 x 100 mL), dried over anhydrous sodium sulfate, concentrated, and dried to obtain a black solid (compound 5), 12.84 g, yield 55.2%.

[0074] (5) Preparation of (1,2,3,4-tetrahydroisoquinoline-1-yl)methylamine hydrochloride (compound 6)

[0075] Compound 5 (12.84 g, 0.044 mol, leq) was dissolved in 100 mL of methanol. 80% hydrazine hydrate (6.4 mL, 0.132 mol, 3 eq) was added at 80 °C with stirring. The reaction was monitored by TLC until complete. After cooling, the flocculent material was filtered off. The filter cake was washed with methanol, adjusted to a weakly alkaline state with saturated sodium bicarbonate solution, extracted with dichloromethane (3 x 100 mL), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a black oily substance (compound 6), 5.51 g, yield 77.3%. This was directly added to the next step with compound 9a.

[0076] Example 2: Synthesis of key intermediate 9

[0077] The synthetic route for key intermediate 9 is as follows:

[0078]

[0079] The reagents and conditions f are: diethyl oxalate, 4 mol / L hydrochloric acid, 90℃;

[0080] The reagents and conditions g are: phosphorus oxychloride, N,N-dimethylformamide, and 90℃;

[0081] The specific preparation method is as follows:

[0082] (1) Preparation of 2,3-dihydroxyquinoxaline (compound 8a)

[0083] In a clean, round-bottomed flask, add o-phenylenediamine (10 g, 0.092 mol, 1 eq), diethyl oxalate (20.27 g, 0.139 mol, 1.5 eq), and 100 mL of 4 mol / L hydrochloric acid. Stir and reflux at 90 °C for 2 h. Monitor the reaction by TLC until complete. Cool to room temperature, add 400 mL of water, and a large amount of solid precipitates. Filter, wash with water, and dry to obtain 14.5 g of a light green solid (compound 8a), yield 96.7%.

[0084] (2) Preparation of 2,3-dichloroquinoxaline (compound 9a)

[0085] Take a dry, round-bottomed flask, add compound 8a (14.5 g, 0.089 mol, 1 eq), 100 mL of N,N-dimethylformamide, and phosphorus oxychloride (25 mL, 0.268 mol, 3 eq), stir, reflux at 90 °C, and react for 2 h. Monitor the reaction by TLC until complete. Cool to room temperature, pour the reaction solution into crushed ice, stir until a large amount of solid precipitates, filter, wash with water, and dry to obtain a white solid (compound 9a), 16.17 g, yield 90.9%.

[0086] Example 3: Synthesis of key intermediate 11

[0087] The synthetic route for key intermediate 11 is as follows:

[0088]

[0089] The reagents and conditions h are: potassium carbonate, N,N-dimethylformamide, and 120℃;

[0090] The reagents and conditions were: 3-bromopropanol, sodium hydride, N,N-dimethylformamide, and 45°C.

[0091] The specific preparation method is as follows:

[0092] (1) Preparation of 4b,5,14,15-tetrahydro-6H-isoquinolino[2',1':1,6]pyrazine[2,3-b]quinoxaline (compound 10)

[0093] Compound 6 (5.51 g, 0.034 mol, 1 eq) and compound 9a (6.76 g, 0.034 mol, 1 eq) were dissolved in 120 mL of N,N-dimethylformamide, and potassium carbonate (9.79 g, 0.136 mol, 4 eq) was added. The mixture was heated to 110 °C and refluxed for 12 h. The reaction was monitored by TLC until complete. Then, the mixture was cooled to room temperature, and a large amount of water was added. The mixture was stirred, and a large amount of solid precipitated. The solid was filtered and dried. The solid was recrystallized from the solid using a mixed solvent (isopropanol:tetrahydrofuran = 1:2), filtered, and dried to give a pink solid (compound 10), 3.31 g, with a yield of 33.8%.

[0094] (2) Preparation of 3-(4b,5,14,15-tetrahydro-6H-isoquinolino[2',1':1,6]pyrazine[2,3-b]quinoxalin-6-yl)-1-propanol (compound 11)

[0095] In a clean, round-bottomed flask, compound 10 (1.96 g, 0.007 mol, 1 eq) and 120 mL of N,N-dimethylformamide were added. Sodium hydride (1.13 g, 0.027 mol, 4 eq) was added under ice bath conditions. After stirring for half an hour, 3-bromopropanol (1.13 g, 0.008 mol, 1.2 eq) was added. The mixture was refluxed at 45 °C, and the reaction was monitored by TLC until completion. After cooling to room temperature, 120 mL of water was added. The mixture was filtered to remove insoluble matter, extracted with dichloromethane (3 x 100 mL), washed with 100 mL of saturated brine, and the organic layers were combined. The mixture was dried over anhydrous sodium sulfate, concentrated, and passed through a column (mobile phase: dichloromethane). The concentrated product yielded a golden-yellow solid (compound 11), 0.91 g, with a yield of 38.7%. 1 HNMR(600MHz,DMSO-d6)δ7.48-7.46(m,1H),7.46-7.43(m,1H),7.41-7.37(m,1H),7.30-7.23(m, 3H),7.22-7.17(m,2H),5.01-4.96(m,1H),4.86(dd,J=10.1,3.6Hz,1H),4.65(s,1H),4.10(dd,J= 11.9,3.7Hz,1H),3.88-3.82(m,1H),3.74-3.68(m,1H),3.50(t,J=6.2Hz,2H),3.49-3.45(m,1H) ,3.08(td,J=12.2,3.5Hz,1H),3.02-2.96m,1H),2.89(dt,J=15.9,3.0Hz,1H),1.89-1.81(m,2H). 13C NMR (151MHz, DMSO-d6) δ143.55,143.29,137.33,136.60,135.44,133.88,129.47,127.44,126. 80,126.16,125.28,124.84,124.68,124.01,58.94,52.67,51.90,45.49,39.64,28.89,28.52.

[0096] Example 4: Synthesis of a five-membered ring (AE ring) TNBG-glucose / galactose conjugate

[0097] The synthetic route for the five-membered ring anticancer drug TNBG-glycoconjugate is as follows:

[0098]

[0099] The reagents and conditions were: 2,3,4,6-tetraacetoxy-α-D-pyranose / galactose bromide, dichloromethane, silver trifluoromethanesulfonate (I), 4A molecular sieve, and room temperature.

[0100] The reagents and conditions k are: potassium carbonate, methanol, and room temperature.

[0101] The specific preparation method is as follows:

[0102] Preparation of (1) (2R,3R,4S,5R,6R)-2-(acetoxymethyl)-6-(3-(4b,5,14,15-tetrahydro-6H-isoquinoline[2',1':1,6]pyrazine[2,3-b]quinoxalin-6-yl)propoxy)tetrahydro-2H-pyran-3,4,5-triacetate (compound 12a)

[0103] Compound 11 (0.66 g, 1.91 mmol, 1 eq) was dissolved in 22 mL of dry dichloromethane. 3 g of 4A molecular sieve was added and the mixture was stirred for 10 min. Then, 2,3,4,6-tetraacetoxy-α-D-glucopyranose bromide (1.25 g, 3.05 mmol, 1 eq) was added. The mixture was stirred for 1 h under nitrogen protection. Silver trifluoromethanesulfonate (I) (0.78 g, 3.05 mmol, 1.6 eq) was added under ice bath. The mixture was stirred overnight at room temperature under nitrogen protection. The reaction was monitored by TLC until completion. The diatomaceous earth was filtered, concentrated, diluted with 20 mL of dichloromethane, and washed successively with water (2 x 20 mL), saturated sodium bicarbonate solution (2 x 20 mL), and saturated brine (20 mL). It was dried over anhydrous sodium sulfate, concentrated, and passed through a column (mobile phase: petroleum ether: ethyl acetate = 1:1). The concentrated product yielded a yellow oil (compound 12a), 0.41 g, with a yield of 31.8%, which was directly used for the next step.

[0104] (2) Preparation of 2R,3S,4S,5R,6R)-2-(acetoxymethyl)-6-(3-(4b,5,14,15-tetrahydro-6H-isoquinoline[2',1':1,6]pyrazine[2,3-b]quinoxalin-6-yl)propoxy)tetrahydro-2H-pyran-3,4,5-triacetate (compound 12b)

[0105] The preparation method is the same as in "Example 4-(1)", and the yellow oil (compound 12b) was concentrated to obtain 0.38 g, with a yield of 43.2%.

[0106] (3) Preparation of (2R,3R,4S,5R,6R)-2-(hydroxymethyl)-6-(3-(4b,5,14,15-tetrahydro-6H-isoquinoline[2',1':1,6]pyrazine[2,3-b]quinoxalin-6-yl)propoxy)tetrahydro-2H-pyran-3,4,5-triol (I-1)

[0107]

[0108] 15 mL of anhydrous methanol was added to compound 12a, the pH was adjusted to 9 with potassium carbonate, and the mixture was stirred at room temperature. The reaction was monitored by TLC until completion. The mixture was filtered through diatomaceous earth, concentrated, and passed through a column (mobile phase: dichloromethane:methanol = 30:1-10:1). The concentrate yielded a pale yellow solid (I-1), 80 mg, yield 26.0%, melting point 129.1-133.1 °C. 1 H NMR (600MHz, DMSO-d6) δ7.53-7.46(m,1H),7.46-7.41(m,2H),7.30-7.23(m,3H),7.22-7.16(m,2H),5.06(dd,J=37.4,4 .8Hz,1H),5.01-4.97(m,1H),4.95(dd,J=4.9,1.6Hz,1H),4.91-4.87(m,2H),4.47(q,J=6.1Hz,1H),4.19(dd,J=10.9,7 .8Hz,1H),4.15-4.11(m,1H),3.94-3.89(m,1H),3.84-3.76(m,2H),3.68-3.63(m,1H),3.60-3.54(m,1H),3.52-3.47(m ,1H),3.45-3.37(m,1H),3.19-3.14(m,1H),3.12-2.96(m,5H),2.92-2.87(m,1H),2.07-1.99(m,1H),1.99-1.91(m,1H). 13CNMR(151MHz,DMSO-d6)δ143.57,143.17,137.47,136.66,135.44,133.96,129.43,127.40,126.83,126.21,125.25,125.06,124 .61,124.01,103.41,77.42,77.20,74.05,70.69,67.02,66.60,61.63,52.72,52.17,45.87,28.54,26.94.HRMS(ESI):m / z[M+H] + Calcd forC 27 H 33 N4O6:509.2400; Found:509.2402.

[0109] (4) Preparation of (2R,3S,4S,5R,6R)-2-(hydroxymethyl)-6-(3-(4b,5,14,15-tetrahydro-6H-isoquinoline[2',1':1,6]pyrazine[2,3-b]quinoxalin-6-yl)propoxy)tetrahydro-2H-pyran-3,4,5-triol (I-2)

[0110]

[0111] The preparation method is the same as in "Example 4-(3)". A pale yellow solid (I-2) was obtained by column chromatography, 110 mg, yield 38.5%, melting point 101.0-103.9℃. 1 HNMR(600MHz,DMSO-d6)δ7.53-7.50(m,1H),7.46-7.40(m,2H),7.30-7.23(m,3H),7.22-7.16(m,2H),5.02-4.97(m,1H),4 .94-4.85(m,2H),4.71(t,J=5.3Hz,1H),4.55-4.49(m,1H),4.36(dd,J=30.6,4.5Hz,1H),4.16-4.10(m,2H),3.93-3.86(m, 1H),3.84-3.72(m,2H),3.65(q,J=3.5Hz,1H),3.59-3.51(m,2H),3.51-3.44(m,2H),3.3-3.33(m,2H),3.31-3.27(m,1H), 3.12-3.05(m,1H),2.99(td,J=13.7,11.5,5.3Hz,1H),2.89(dt,J=15.9,3.0Hz,1H),2.07-1.99(m,1H),1.98-1.90(m,1H). 13C NMR(151MHz,DMSO-d6)δ143.56,143.17,137.47,136.65,135.33,133.93,129.40,127.40,126.93,126.49,126.30,125.24,124 .59,123.99,103.89,75.75,73.95,71.11,68.61,66.84,66.45,60.87,52.69,52.30,45.65,28.53,26.92.HRMS(ESI):m / z[M+H] + Calcd forC 27 H 33 N4O6:509.2400; Found:509.2402.

[0112] Secondly, the synthesis of compounds of formula IV:

[0113] Example 5: Synthesis of key intermediate 15

[0114] The synthetic route for key intermediate 15 is as follows:

[0115]

[0116] The reagents and conditions h are: potassium carbonate, N,N-dimethylformamide, and 120℃;

[0117] The reagents and conditions were: 3-bromopropanol, sodium hydride, N,N-dimethylformamide, and 45°C.

[0118] The specific preparation method is as follows:

[0119] (1) Preparation of 2,3,4,4a,5,6-hexahydro-1H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxaline (compound 14a)

[0120] Compound 9a (16.60 g, 0.083 mol, 1 eq) was dissolved in 200 mL of N,N-dimethylformamide. Compound 13 (14.29 g, 0.125 mol, 1.5 eq) and potassium carbonate (34.58 g, 0.250 mol, 3 eq) were added sequentially. The mixture was heated to 110 °C and refluxed for 3 h. The reaction was monitored by TLC until complete. After cooling to room temperature, a large amount of water was added, and the mixture was stirred. A large amount of solid precipitated, which was filtered and dried. Recrystallization was performed using a mixed solvent (dichloromethane:methanol = 1:2), filtered, and dried to finally obtain a pale yellow solid (compound 14a), 19.99 g, yield 99.8%. 1 H NMR (600MHz, Chloroform- d)δ7.58-7.54(m,1H),7.44-7.39(m,1H),7.28-7.22(m,2H),6.65(s,1H),4.95-4.89(m,1H),3.57-3.50(m,1H),3.41-3.32(m,2H),2.7 2(td,J=13.0,3.1Hz,1H),1.91-1.84(m,2H),1.81-1.77(m,1H),1.62-1.54(m,4.3,3.0Hz,1H),1.53-1.45(m,1H),1.41-1.34(m,1H). 13 C NMR (151MHz, Chloroform-d) δ144.24,144.17,137.77,136.78,125.68,124.69,124.30,124.06,52.99,45.35,44.33,30.09,24.53,23.38.

[0121] (2) Preparation of 9-chloro-2,3,4,4a,5,6-hexahydro-1H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxaline (compound 14f)

[0122] The preparation method was the same as in Example 5-(1), and a pale purple-red solid (compound 14f) was obtained by column chromatography with a yield of 40.8%. 1 H NMR(600MHz,Chloroform-d)δ7.46(d,J=8.7Hz,1H),7.40(d,J=2.4Hz,1H),7.1 9(dd,J=8.7,2.4Hz,1H),5.96(s,1H),4.91-4.86(m,1H),3.57-3.52(m,1H),3.4 2-3.36(m,2H),2.73(td,J=13.0,3.0Hz,1H),1.94-1.90(m,1H),1.89-1.85(m,1 H),1.84-1.80(m,1H),1.62-1.55(m,1H),1.55-1.47(m,1H),1.43-1.36(m,1H). 13 C NMR (151MHz, Chloroform-d) δ144.16,144.08,137.25,136.28,129.68,126.58,124.84,123.44,52.91,45.34,44.31,30.05,24.46,23.30.

[0123] (3) The preparation method of intermediate 14b-14e is the same as that in “Example 5-(1)”.

[0124] (4) Preparation of 2-(1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)-1-ethanol (compound 15a)

[0125] Compound 14a (2.50 g, 0.010 mol, 1 eq) and 70 mL of N,N-dimethylformamide were added to a clean, round-bottom flask. 60% sodium hydride (1.66 g, 0.042 mol, 4 eq) was added under ice bath conditions. After stirring for half an hour, 2-bromoethanol (1.95 g, 0.016 mol, 1.5 eq) was added. The mixture was refluxed at 45 °C, and the reaction was monitored by TLC until completion. After cooling to room temperature, 100 mL of water was added. The mixture was filtered to remove insoluble matter, washed with dichloromethane (3 x 100 mL) and saturated brine, and the organic layers were combined. The mixture was dried over anhydrous sodium sulfate, concentrated, and passed through a column (mobile phase: dichloromethane). The concentrated product yielded a pale yellow solid (compound 15a), 1.80 g, with a yield of 60.9%. 1 H NMR(600MHz,Chloroform-d)δ7.56-7.52(m,1H),7.50-7.45m,1H),7.25-7.22(m ,2H),5.42(s,1H),4.87-4.83m,1H),3.96-3.89m,2H),3.83-3.76(m,2H),3.46( d,J=6.3Hz,2H),3.39-3.33(m,1H),2.70(td,J=13.0,3.1Hz,1H),1.93-1.84(m, 2H),1.82-1.77(m,1H),1.62-1.53(m,1H),1.53-1.45(m,1H),1.38-1.30m,1H). 13 C NMR(151MHz,Chloroform-d)δ144.29,144.29,137.15,135.99,125.43,124. 87,124.51,124.34,62.67,53.67,53.36,52.38,44.44,29.93,24.39,23.22.

[0126] (5) Preparation of 3-(1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)-1-propanol (compound 15b)

[0127] The preparation method was the same as in "Example 5-(4)". The pale yellow solid (compound 15b) was obtained by column chromatography with a yield of 74.1%. 1H NMR(600MHz,Chloroform-d)δ7.56-7.52(m,1H),7.48-7.44(m,1H),7.26-7.21m,2H) ,5.48(s,1H),4.88-4.82(m,1H),3.90-3.85(m,1H),3.80-3.74(m,1H),3.58-3.53(m ,1H),3.53-3.48(m,1H),3.43-3.38(m,1H),3.37-3.30(m,2H),2.71(td,J=13.1,3.1 Hz,1H),1.94-1.78(m,5H),1.64-1.56(m,1H),1.54-1.46(m,1H),1.39-1.32(m,1H). 13 CNMR(151MHz,Chloroform-d)δ143.88,143.73,136.80,136.26,125.50,125.06, 124.26,124.07,57.66,52.07,51.33,44.66,44.38,30.18,29.99,24.31,23.25.

[0128] (6) The preparation method of intermediate 15c-15l is the same as that in “Example 5-(4)”.

[0129] Example 6: Synthesis of a four-membered ring (ABCE ring) TNBG-glucose / galactose conjugate

[0130] The synthetic route for the four-membered ring (ABCE ring) anticancer drug TNBG-glycoconjugate is as follows:

[0131]

[0132] The reagents and conditions were: 2,3,4,6-tetraacetoxy-α-D-pyranose / galactose bromide, dichloromethane, silver trifluoromethanesulfonate (I), 4A molecular sieve, and room temperature.

[0133] The reagents and conditions k are: potassium carbonate, methanol, and room temperature.

[0134] The specific preparation method is as follows:

[0135] Preparation of (1) (2R,3R,4S,5R,6R)-2-(acetoxymethyl)-6-(2-(1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)tetrahydro-2H-pyran-3,4,5-triacetate (compound 16a)

[0136] Compound 15a (1.3 g, 4.57 mmol, 1 eq) was dissolved in 18 mL of dry dichloromethane. 0.36 g of activated 4A molecular sieve was added and the mixture was stirred for 10 min. Then, 2,3,4,6-tetraacetoxy-α-D-pyranoselure bromide (2.26 g, 5.49 mmol, 1.2 eq) was added. Under nitrogen protection, the mixture was stirred for 1 h. Silver trifluoromethanesulfonate (I) (1.41 g, 5.49 mmol, 1.2 eq) was added under ice bath. Under nitrogen protection, the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until completion. The diatomaceous earth was filtered, concentrated, diluted with 20 mL of dichloromethane, and washed successively with water (2 x 20 mL), saturated sodium bicarbonate solution (2 x 20 mL), and saturated brine (20 mL). It was dried over anhydrous sodium sulfate, concentrated, and passed through a column (mobile phase: petroleum ether: ethyl acetate = 1:1) to obtain a pale yellow solid (compound 16a), 0.63 g, yield 22.4%, which was directly added to the next step.

[0137] (2) Preparation of (2R,3S,4S,5R,6R)-2-(acetoxymethyl))-6-(3-(9-chloro-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)tetrahydro-2H-pyran-3,4,5-triacetate (compound 16x)

[0138] The preparation method is the same as in "Example 6-(1)". A pale yellow solid (compound 16x) was obtained by column chromatography, 0.90 g, yield 86.5%, and proceeded directly to the next step.

[0139] (3) The preparation method of compound 16b-16w is the same as that in “Example 6-(1)”.

[0140] (4) Preparation of (2R,3R,4S,5R,6R)-2-(2-(1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-1)

[0141]

[0142] Compound 16a (0.63 g, 1.02 mmol, 1 eq) was dissolved in 15 mL of anhydrous methanol, the pH was adjusted to 9 with potassium carbonate, and the mixture was stirred at room temperature. The reaction was monitored by TLC until completion. The solution was filtered through diatomaceous earth, concentrated, and passed through a column (mobile phase: dichloromethane:methanol = 30:1-10:1). The concentrate yielded a white solid (IV-1), 150 mg, yield 32.8%, melting point 94.1-96.0 °C. 1H NMR (600MHz, DMSO-d6) δ7.39(dd,J=12.8,7.8Hz,2H),7.21-7.13(m,2H),5.03(t,J=5.1Hz,1H),4.93(t,J=4.4Hz,1H ),4.88(d,J=5.3Hz,1H),4.68(d,J=13.3Hz,1H),4.48-4.41(m,1H),4.19(d,J=7.8Hz,1H),4.04-3.97(m,1H),3.94- 3.86(m,1H),3.79-3.71(m,2H),3.69-3.57(m,2H),3.54-3.39(m,3H),3.15-3.12(m,1H),3.11-3.07(m,1H),3.07-3 .02(m,1H),2.97(q,J=7.5Hz,1H),2.67(t,J=12.1Hz,1H),1.84-1.76(m,3H),1.50-1.42(m,2H),1.30-1.25(m,1H). 13 C NMR(151MHz,DMSO-d6)δ144.32,143.23,137.32,136.82,125.37,124.93,124.63,123.94,103.64,77.36, 77.17,73.95,70.55,66.65,61.55,52.15,51.89,47.89,44.44,29.61,24.44,23.22.HRMS(ESI):m / z[M+H] + Calcd for C 22 H 31 N4O6:447.2244; Found:447.2245.

[0143] (5) Preparation of (2R,3S,4S,5R,6R)-2-(2-(1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-2)

[0144]

[0145] The preparation method is the same as in "Example 6-(4)". The white solid (IV-2) was obtained by column chromatography, 60 mg, yield 18.4%, melting point 51.8-54.1℃. 1H NMR(600MHz,DMSO-d6)δ7.42-7.36(m,2H),7.20-7.14(m,2H),4.89-4.86m,1H),4.70-4.65(m,2H),4. 54-4.50(m,1H),4.35-4.32(m,1H),4.13(d,J=7.4Hz,1H),4.02-3.96(m,1H),3.92-3.84(m,1H),3.81

[0146] -3.64(m,3H),3.64-3.61(m,1H),3.54-3.41(m,3H),3.39-3.34(m,2H),3.31-3.28(m,1H),3. 28-3.24(m,1H),2.71-2.63(m,1H),1.84-1.76(m,3H),1.50-1.41(m,2H),1.29-1.22(m,1H). 13 C NMR(151MHz,DMSO-d6)δ144.32,143.20,137.32,136.81,125.37,124.93,124.62,123.91,104.25,75.65, 73.89,71.06,68.59,66.52,60.83,52.11,51.91,47.95,44.42,29.62,24.43,23.21.HRMS(ESI):m / z[M+H] + Calcd for C 22 H 31 N4O6:447.2244; Found:447.2248.

[0147] (6) Preparation of (2R,3R,4S,5R,6R)-2-(3-(1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)-(6-hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-3)

[0148]

[0149] The preparation method is the same as in "Example 6-(4)". The white solid (IV-3) was obtained by column chromatography, 450 mg, yield 43.9%, melting point 105.6-108.3℃. 1H NMR(600MHz,DMSO-d6)δ7.39(d,J=7.6Hz,2H),7.21-7.12(m,2H),5.02(dd,J=12.9,4.8Hz,1H),4.99-4.86(m,2H),4.68 (d,J=13.3Hz,1H),4.48(t,J=6.0Hz,1H),4.15(dd,J=7.8,2.9Hz,1H),3.91-3.81(m,1H),3.72-3.64(m,3H),3.63-3.57( m,1H),3.56-3.50(m,1H),3.46-3.41(m,1H),3.40-3.36(m,2H),3.15(t,J=8.8Hz,1H),3.12-3.08(m,1H),3.08-3.03(m ,1H),3.02-2.95(m,1H),2.71-2.63(m,1H),1.98-1.87(m,2H),1.84-1.75(m,3H),1.50-1.41(m,2H),1.29-1.24(m,1H). 13 C NMR (151MHz, DMSO-d6) δ144.27,143.34,137.45,136.77,125.33,124.94,124.57,123.85,103.32,77.35,77. 19,73.97,70.58,66.77,61.61,52.15,51.12,45.25,44.40,29.68,26.87,24.40,23.23.HRMS(ESI):m / z[M+H] + Calcd for C 23 H 33 N4O6:461.2400; Found:461.2403.

[0150] (7) Preparation of (2R,3S,4S,5R,6R)-2-(3-(1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)-6-hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-4)

[0151]

[0152] The preparation method is the same as in "Example 6-(4)". A pale yellow solid (IV-4) was obtained by column chromatography, 115 mg, yield 39.2%, melting point 150.2-152.9℃. 1HNMR(600MHz,DMSO-d6)δ7.42-7.36(m,2H),7.21-7.11(m,2H),4.86(dd,J=11.9,4.6Hz,1H),4.70(d,J=5.4Hz,1 H),4.70-4.65(m,1H),4.58-4.52(m,1H),4.35(dd,J=4.6,2.8Hz,1H),4.10(dd,J=7.4,3.8Hz,1H),3.86-3.80(m, 1H),3.75-3.59(m,4H),3.56-3.49(m,2H),3.49-3.45(m,1H),3.41-3.34(m,3H),3.32-3.29(m,1H),3.29-3.26( m,1H),2.67(td,J=12.8,3.2Hz,1H),1.97-1.86(m,2H),1.85-1.75(m,3H),1.51-1.42(m,2H),1.30-1.24(m,1H). 13 C NMR (151MHz, DMSO-d6) δ144.26,143.34,137.46,136.76,125.33,124.95,124.57,123.83,103.81,75.65,73. 92,71.06,68.59,66.37,60.89,52.16,51.20,45.25,44.39,29.66,26.93,24.41,23.23.HRMS(ESI):m / z[M+H] + Calcd for C 23 H 33 N4O6:461.2400; Found:461.2404.

[0153] (8) Preparation of (2R,3R,4S,5R,6R)-2-(2-(9,10-dimethyl-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-5)

[0154]

[0155] The preparation method is the same as in "Example 6-(4)". The yellow solid (IV-5) was obtained by column chromatography, 160 mg, yield 27.8%, melting point 159.6-162.4℃. 1HNMR (600MHz, DMSO-d6) δ7.20 (s, 1H), 7.19 (s, 1H), 5.03 (d, J = 5.8Hz, 1H), 4.99-4.91 (m, 1H), 4.91-4.86 (m, 1H), 4. 65-4.60(m,1H),4.46(d,J=11.9Hz,1H),4.18(d,J=7.8Hz,1H),4.03-3.96(m,1H),3.90-3.80(m,1H),3.80-3.69(m, 2H),3.69-3.57(m,2H),3.49-3.40(m,2H),3.32-3.28(m,1H),3.14(t,J=8.8Hz,1H),3.11-3.07(m,1H),3.07-3.02( m,1H),2.99-2.94(m,1H),2.66-2.59(m,1H),2.26(s,6H),1.83-1.74(m,3H),1.48-1.39(m,2H),1.27-1.22(m,1H). 13 C NMR(151MHz,DMSO-d6)δ143.96,142.85,135.46,134.96,133.42,132.63,125.36,124.92,103.47,77.10,76.84, 73.77,70.36,66.65,61.37,55.20,52.17,47.81,44.45,29.57,24.38,23.16,19.79,19.76.HRMS(ESI):m / z[M+H] + Calcd for C 24 H 35 N4O6:475.2557; Found:475.2557.

[0156] (9) Preparation of (2R,3S,4S,5R,6R)-2-(2-(9,10-dimethyl-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-6)

[0157]

[0158] The preparation method is the same as in "Example 6-(4)". A pale yellow solid (IV-6) was obtained by column chromatography, 140 mg, yield 23.4%, melting point 216.6-219.1℃. 1HNMR(600MHz,DMSO-d6)δ7.20(s,1H),7.19(s,1H),4.87(t,J=4.4Hz,1H),4.71-4.66(m,1H),4.66-4 .60(m,1H),4.55-4.49(m,1H),4.36-4.32(m,1H),4.12(dd,J=7.4,1.1Hz,1H),4.01-3.94(m,1H),3. 88-3.80(m,1H),3.79-3.72(m,1H),3.72-3.67(m,1H),3.64-3.57(m,2H),3.54-3.40(m,3H),3.33-3 .24(m,4H),2.66-2.59(m,1H),2.26(s,6H),1.82-1.74(m,3H),1.48-1.39(m,2H),1.27-1.19(m,1H). 13 C NMR(151MHz,DMSO-d6)δ143.95,142.85,135.59,135.05,133.29,132.47,125.47,125.05,104.25,75.65,73.87, 71.07,68.59,66.51,60.83,55.38,52.15,47.94,44.38,29.58,24.47,23.23,19.83,19.80.HRMS(ESI):m / z[M+H] + Calcd for C 24 H 35 N4O6:475.2557; Found:475.2560.

[0159] (10) Preparation of (2R,3R,4S,5R,6R)-2-(3-(9,10-dimethyl-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-7)

[0160]

[0161] The preparation method is the same as in "Example 6-(4)". The white solid (IV-7) was obtained by column chromatography, 190 mg, yield 42.6%, melting point 116.8-119.2℃. 1HNMR(600MHz,DMSO-d6)δ7.21(s,1H),7.19(s,1H),5.00(dd,J=13.6,4.8Hz,1H),4.97-4.83(m,2H),4.64(d,J=1 2.7Hz,1H),4.50-4.41(m,1H),4.15(dd,J=7.8,2.6Hz,1H),3.88-3.81(m,1H),3.70-3.63(m,3H),3.63-3.48(m, 3H),3.47-3.48(m,2H),3.17-3.13(m,1H),3.12-3.07(m,1H),3.07-3.02(m,1H),2.97(t,J=8.7Hz,1H),2.66-2. 60(m,1H),2.26(s,3H),2.25(s,3H),1.95-1.84(m,2H),1.82-1.75(m,3H),1.48-1.42(m,2H),1.29-1.23(m,1H). 13 C NMR (151MHz, DMSO-d6) δ143.87,143.01,135.75,135.03,133.22,132.38,125.45,125.08,103.21,77.33,77.21,73. 97,70.59,66.65,61.61,52.18,51.27,45.24,44.36,29.60,26.87,24.45,23.26,19.82,19.80.HRMS(ESI):m / z[M+H] + Calcd for C 25 H 37 N4O6:489.2713; Found:489.2717.

[0162] (11) Preparation of (2R,3S,4S,5R,6R)-2-(3-(9,10-dimethyl-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-8)

[0163]

[0164] The preparation method is the same as in "Example 6-(4)". The white solid (IV-8) was obtained by column chromatography, 50 mg, yield 14.6%, melting point 186.9-190.0℃. 1HNMR(600MHz,DMSO-d6)δ7.20(s,1H),7.19(s,1H),4.83(s,1H),4.73-4.61(m,2H),4.54(s,1H) ),4.38-4.28(m,1H),4.10(dd,J=7.3,3.4Hz,1H),3.85-3.80(m,1H),3.70-3.60(m,3H),3.60-3 .56(m,1H),3.55-3.50(m,2H),3.49-3.45(m,1H),3.32-3.25(m,5H),2.66-2.60(m,1H),2.26(s ,3H),2.25(s,3H),1.95-1.85(m,2H),1.83-1.75(m,3H),1.50-1.40(m,2H),1.29-1.24(m,1H). 13 C NMR (151MHz, DMSO-d6) δ143.89,142.99,135.55,134.88,133.36,132.55,125.31,124.93,103.78,75.46,73.68,70. 95,68.47,66.81,60.80,52.22,51.24,45.20,44.44,29.62,26.85,24.37,23.18,19.77,19.74.HRMS(ESI):m / z[M+H] + Calcd for C 25 H 37 N4O6:489.2713; Found:489.2716.

[0165] (12) Preparation of (2R,3R,4S,5R,6R)-2-(2-(9,10-difluoro-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-9)

[0166]

[0167] The preparation method is the same as in "Example 6-(4)". A pale yellow solid (IV-9) was obtained by column chromatography, 100 mg, yield 14.9%, melting point 114.8-117.6℃. 1HNMR (600MHz, DMSO-d6) δ7.37-7.33(m,1H),7.33-7.29(m,1H),5.03(t,J=4.7Hz,1H),4.94(t,J=4.3Hz,1H),4.89(d,J=5.3Hz ,1H),4.62(d,J=13.3Hz,1H),4.48-4.42(m,1H),4.18(d,J=7.8Hz,1H),4.02-3.95(m,1H),3.92-3.84(m,1H),3.81-3.75(m,1 H),3.74-3.70(m,1H),3.70-3.62(m,2H),3.52-3.43(m,1H),3.43-3.40(m,1H),3.39-3.36(m,1H),3.15-3.11(m,1H),3.11-3 .07(m,1H),3.06-3.01(m,1H),2.99-2.93(m,1H),2.71-2.64(m,1H),1.84-1.74m,3H),1.49-1.40(m,2H),1.28-1.21(m,1H). 13 CNMR(151MHz,DMSO-d6)δ147.95,146.36,144.49,143.39,133.94,133.37,111.61,111.15,103.46,77.36, 77.15,73.93,70.55,66.51,61.57,52.00,51.70,47.95,44.34,29.55,24.36,23.11.HRMS(ESI):m / z[M+H] + Calcd for C 22 H 29 N4O6F2:483.2055; Found:483.2059.

[0168] (13) Preparation of (2R,3S,4S,5R,6R)-2-(2-(9,10-difluoro-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-10)

[0169]

[0170] The preparation method is the same as in "Example 6-(4)". The white solid (IV-10) was obtained by column chromatography, 100 mg, yield 16.4%, melting point 245.3-248.0℃. 1HNMR(600MHz,DMSO-d6)δ7.36-7.33(m,1H),7.33-7.28(m,1H),4.87(t,J=4.4Hz,1H),4.74-4.66(m,1H),4.65-4.60(m,1H) ,4.52(t,J=5.7Hz,1H),4.37-4.30(m,1H),4.13(d,J=7.3Hz,1H),4.01-3.94(m,1H),3.90-3.82(m,1H),3.79-3.74(m,1H),3 .74-3.70(m,1H),3.70-3.65(m,1H),3.64-3.61(m,1H),3.54-3.46(m,2H),3.45-3.41(m,1H),3.40-3.35(m,1H),3.32-3.3 0(m,1H),3.30-3.27(m,1H),3.27-3.24(m,1H),2.71-2.64(m,1H),1.83-1.75(m,3H),1.49-1.40(m,2H),1.29-1.20(m,1H). 13 C NMR(151MHz,DMSO-d6)δ147.90,146.30,144.48,143.38,133.74,133.21,111.56,111.10,103.79,75.39, 73.54,70.87,68.43,66.29,60.73,51.91,51.65,47.96,44.41,29.50,24.26,22.94.HRMS(ESI):m / z[M+H] + Calcd for C 22 H 29 N4O6F2:483.2055; Found:483.2059.

[0171] (14) Preparation of (2R,3R,4S,5R,6R)-2-(3-(9,10-difluoro-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-11)

[0172]

[0173] The preparation method is the same as in "Example 6-(4)". The white solid (IV-11) was obtained by column chromatography, 90 mg, yield 17.3%, melting point 163.4-165.8℃. 1HNMR (600MHz, DMSO-d6) δ7.36-7.28(m,2H),5.01(dd,J=12.9,4.9Hz,1H),4.94(dd,J=4.8,1.0Hz,1H),4.89(d,J=5.1Hz,1H),4. 63(dd,J=13.2,2.6Hz,1H),4.46(t,J=5.8Hz,1H),4.14(dd,J=7.8,3.7Hz,1H),3.87-3.82(m,1H),3.73-3.66(m,2H),3.66-3.63 (m,1H),3.63-3.59(m,1H),3.54-3.49(m,1H),3.46-3.38(m,2H),3.38-3.34(m,1H),3.17-3.12(m,1H),3.11-3.07(m,1H),3.06 -3.02(m,1H),3.00-2.95(m,1H),2.71-2.65(m,1H),1.96-1.84(m,2H),1.83-1.75(m,3H),1.50-1.40(m,2H),1.29-1.22(m,1H). 13 CNMR(151MHz,DMSO-d6)δ147.89,146.30,144.46,143.54,134.08,133.32,111.67,111.26,103.30,77.36,77. 20,73.97,70.60,66.66,61.62,52.00,50.93,45.30,44.37,29.64,26.81,24.33,23.13.HRMS(ESI):m / z[M+H] + Calcd for C 23 H 31 N4O6F2:497.2212; Found:497.2215.

[0174] Preparation of (15) (2R,3S,4S,5R,6R)-2-(3-(9,10-difluoro-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-12)

[0175]

[0176] The preparation method is the same as in "Example 6-(4)". The white solid (IV-12) was obtained by column chromatography, 92 mg, yield 20.5%, melting point 108.2-111.2℃. 1HNMR(600MHz,DMSO-d6)δ7.35-7.30(m,2H),4.86(dd,J=12.4,4.6Hz,1H),4.70(d,J=5.4Hz,1H) ,4.66-4.60(m,1H),4.56-4.52(m,1H),4.34(dd,J=4.6,2.9Hz,1H),4.09(dd,J=7.4,4.3Hz,1H) ,3.86-3.79(m,1H),3.74-3.59(m,4H),3.55-3.44(m,3H),3.42-3.35(m,2H),3.33-3.25(m,3H) ,2.71-2.65m,1H),1.95-1.85(m,2H),1.84-1.76(m,3H),1.51-1.40(m,2H),1.28-1.22(m,1H). 13 C NMR (151MHz, DMSO-d6) δ147.85,146.16,144.40,143.49,133.87,133.06,111.39,111.08,103.62,75.35,73. 56,70.93,68.45,66.43,60.79,51.95,50.86,45.28,44.45,29.54,26.67,24.22,22.98.HRMS(ESI):m / z[M+H] + Calcd for C 23 H 30 N4O6F2Na:519.2031; Found:519.2033.

[0177] Preparation of (16) (2R,3R,4S,5R,6R)-2-(2-(9,10-dichloro-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-13)

[0178]

[0179] The preparation method is the same as in "Example 6-(4)". A yellow solid (IV-13) was obtained by column chromatography, 210 mg, yield 34.4%, melting point 141.2-144.1℃. 1HNMR (600MHz, DMSO-d6) δ7.54(s,1H),7.52(s,1H),5.03(dd,J=5.1,3.3Hz,1H),4.94(dd,J=4.9,3.7Hz,1H) ,4.89(dd,J=5.3,1.0Hz,1H),4.67-4.62(m,1H),4.47-4.43(m,1H),4.18(dd,J=7.8,1.1Hz,1H),4.02-3.96( m,1H),3.93-3.86(m,1H),3.82-3.73(m,2H),3.73-3.63(m,2H),3.46-3.38(m,3H),3.15-3.07(m,2H),3.06- 3.01(n,1H),2.98-2.93(m,1H),2.74-2.66(m,1H),1.85-1.76(m,3H),1.48-1.41(m,2H),1.28-1.23(m,1H). 13 CNMR(151MHz,DMSO-d6)δ144.88,143.78,136.85,136.44,125.97,125.61,125.26,125.21,103.41,77.12, 76.82,73.74,70.37,66.49,61.38,51.99,51.57,48.08,44.43,29.49,24.29,22.97.HRMS(ESI):m / z[M+H] + Calcd for C 22 H 29 N4O6Cl2:515.1464; Found:515.1468.

[0180] Preparation of (17) (2R,3S,4S,5R,6R)-2-(2-(9,10-dichloro-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-14)

[0181]

[0182] The preparation method is the same as in "Example 6-(4)". A pale yellow solid (IV-14) was obtained by column chromatography, 110 mg, yield 22.4%, melting point 206.8-209.7℃. 1HNMR (600MHz, DMSO-d6) δ7.54(s,1H),7.52(s,1H),4.87(dd,J=4.8,3.3Hz,1H),4.69(dd,J=5.3,1.5Hz,1 H),4.67-4.62(m,1H),4.53-4.50(m,1H),4.34(t,J=4.3Hz,1H),4.13(d,J=7.3Hz,1H),4.02-3.95(m,1H) ,3.89-3.83(m,1H),3.80-3.67(m,3H),3.64-3.60(m,1H),3.54-3.45(m,2H),3.44-3.39(m,2H),3.32-3. 27(m,2H),3.27-3.24(m,1H),2.73-2.66(m,1H),1.84-1.76(m,3H),1.49-1.40(m,2H),1.29-1.23(m,1H). 13 CNMR(151MHz,DMSO-d6)δ144.91,143.80,136.97,136.52,125.95,125.70,125.29,125.21,104.18,75.63, 73.85,71.04,68.56,66.34,60.80,51.97,51.67,48.16,44.37,29.53,24.34,23.04.HRMS(ESI):m / z[M+H] + Calcd for C 22 H 28 N4O6Cl2Na:537.1284; Found:537.1288.

[0183] Preparation of (18) (2R,3R,4S,5R,6R)-2-(3-(9,10-dichloro-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-15)

[0184]

[0185] The preparation method is the same as in "Example 6-(4)". The light yellow solid (IV-15) was obtained by column chromatography, 260 mg, yield 31.3%, melting point 145.7-148.5℃. 1H NMR (600MHz, DMSO-d6) δ7.54(d,J=1.3Hz,2H),5.03(dd,J=14.0,4.9Hz,1H),4.97-4.93(m,1H),4.91(d,J=5. 2Hz,1H),4.68-4.63(m,1H),4.47(t,J=5.9Hz,1H),4.14(dd,J=7.8,4.0Hz,1H),3.87-3.81(m,1H),3.74-360 (m,4H),3.54-3.49(m,1H),3.48-3.39(m,3H),3.17-3.12(m,1H),3.11-3.07(m,1H),3.06-3.01(m,1H),3.00 -2.95(m,1H),2.73-2.68(m,1H),1.95-1.86(m,2H),1.85-1.76(m,3H),1.49-1.41(m,2H),1.29-1.24(m,1H). 13 C NMR (151MHz, DMSO-d6) δ144.84,143.95,137.11,136.50,125.91,125.65,125.31,125.14,103.31,77.36,77. 19,73.96,70.59,66.65,61.61,52.00,50.85,45.48,44.37,29.64,26.78,24.33,23.06.HRMS(ESI):m / z[M+H] + Calcd for C 23 H 31 N4O6Cl2:529.1621; Found:529.1622.

[0186] Preparation of (19) (2R,3S,4S,5R,6R)-2-(3-(9,10-dichloro-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (IV-16)

[0187]

[0188] The preparation method is the same as in "Example 6-(4)". A yellow solid (IV-16) was obtained by column chromatography, 40 mg, yield 11.8%, melting point 106.8-109.6℃. 1H NMR(600MHz,DMSO-d6)δ7.53(d,J=0.9Hz,2H),4.85(dd,J=13.4,4.6Hz,1H),4.70(d,J=5.3 Hz,1H),4.66(dd,J=13.8,3.5Hz,1H),4.55-4.52(m,1H),4.34(dd,J=4.6,2.9Hz,1H),4.10( dd,J=7.4,4.4Hz,1H),3.85-3.80(m,1H),3.76-3.59(m,4H),3.56-3.49(m,2H),3.49-3.42 (m,2H),3.40-3.35(m,1H),3.34(m,1H),3.31-3.25(m,2H),2.74-2.68(m,1H),1.96-1.85(m 2H),1.85-1.76(m,3H),1.51-1.40(m,2H),1.30-1.23(m,1H). 13 C NMR (151MHz, DMSO-d6) δ144.87,143.94,137.12,136.49,125.91,125.66,125.30,125.12,103.94,75.66,73. 93,71.06,68.59,66.57,60.90,52.01,50.92,45.49,44.35,29.56,26.83,24.33,23.05.HRMS(ESI):m / z[M+H] + Calcd forC 23 H 30 N4O6Cl2Na:551.1440; Found:551.1443.

[0189] Preparation of (20) (2R,3R,4S,5R,6R)-2-(hydroxymethyl)-6-(2-(9-methoxy-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)tetrahydro-2H-pyran-3,4,5-triol (IV-17)

[0190]

[0191] The preparation method is the same as in "Example 6-(4)". The white solid (IV-17) was obtained by column chromatography, 88 mg, yield 17.8%, melting point 181.9-183.9℃. 1HNMR (600MHz, DMSO-d6) δ7.32(dd,J=8.8,0.8Hz,1H),6.87(d,J=2.8Hz,1H),6.80(dd,J=8.8,2.8Hz,1H),5.04(t,J=4.7Hz,1H),4.94(t,J=4. 4Hz,1H),4.89(dd,J=5.3,1.5Hz,1H),4.60-4.55(m,1H),4.47-4.43(m,1H),4.19(dd,J=7.8,1.6Hz,1H),4.04-3.97(m,1H),3.95-3.84(m,1H ),3.82-3.79(m,1H),3.78(s,3H),3.77-3.73(m,1H),3.73-3.67(m,1H ),3.67-3.62(m,1H),3.52-3.40(m,2H),3.31-3.25(m,1H),3.16-3.11 (m,1H),3.11-3.07(m,1H),3.07-3.02(m,1H),2.99-2.95(m,1H),2.65 -2.57(m,1H),1.86-1.72(m,3H),1.50-1.38(m,2H),1.28-1.19(m,1H). 13 C NMR (151MHz, DMSO-d6) δ157.10,143.44,143.05,138.31,131.32,126.26,113.90,105.98,103.64,77.37,77. 17,73.95,70.56,66.67,61.56,55.67,52.19,52.03,47.82,44.50,29.53,24.44,23.20.HRMS(ESI):m / z[M+H] + Calcd for C 23 H 33 N4O7:477.2349; Found:477.2353.

[0192] (21) Preparation of (2R,3S,4S,5R,6R)-2-(hydroxymethyl)-6-(2-(9-methoxy-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)tetrahydro-2H-pyran-3,4,5-triol (IV-18)

[0193]

[0194] The preparation method is the same as in "Example 6-(4)". The white solid (IV-18) was obtained by column chromatography, 100 mg, yield 24.2%, melting point 182.9-185.4℃. 1 H NMR (600MHz, DMSO-d6) δ7.32(d,J=8.8Hz,1H),6.87(d,J=2.8Hz,1H),6.80(dd,J=8.8,2.8Hz,1H),4.89(t,J=4.4Hz,1H) ,4.70(dd,J=5.4,2.2Hz,1H),4.60-4.55(m,1H),4.55-4.51(m,1H),4.35(dd,J=4.6,2.9Hz,1H),4.13(d,J=7.4Hz,1H),4 .03-3.95(m,1H),3.93-3.83(m,1H),3.78(s,3H),3.80-3.78(m,1H),3.77-3.73(m,1H),3.73-3.68(m,1H),3.64-3.61(m ,1H),3.54-3.41(m,3H),3.33-3.24(m,4H),2.64-2.57(m,1H),1.83-1.74(m,3H),1.49-1.38(m,2H),1.27-1.19(m,1H). 13 C NMR (151MHz, DMSO-d6) δ157.04,143.47,143.11,138.11,131.18,126.18,113.83,105.88,103.97,75.40,73. 55,70.88,68.45,66.56,60.75,55.65,52.10,51.98,47.87,44.55,29.46,24.31,23.07.HRMS(ESI):m / z[M+H] + Calcd for C 23 H 33 N4O7:477.2349; Found:477.2351.

[0195] (22) Preparation of (2R,3R,4S,5R,6R)-2-(hydroxymethyl)-6-(3-(9-methoxy-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)tetrahydro-2H-pyran-3,4,5-triol (IV-19)

[0196]

[0197] The preparation method is the same as in "Example 6-(4)". A yellow solid (IV-19) was obtained by column chromatography, 140 mg, yield 21.4%, melting point 101.8-104.7℃. 1 H NMR (600MHz, DMSO-d6) δ7.31(d,J=8.8Hz,1H),6.87(d,J=2.8Hz,1H),6.79(dd,J=8.8,2.8Hz,1H),5.01(dd,J=12.6,4.9Hz,1H),4.93(dd, J=4.9,1.0Hz,1H),4.89(d,J=5.2Hz,1H),4.61-4.57(m,1H),4.46(t,J=5.9Hz,1H),4.16(dd,J=7.8,3.4Hz,1H),3.88-3.83(m,1H),3.78( s,3H),3.72-3.65(m,3H),3.61-3.56(m,1H),3.55-3.50(m,1H),3.48-3.40(m,2H),3.32-3.28(m,1H),3.17-3.13(m,1H),3.12-3.08(m,1 H),3.07-3.02(m,1H),3.01-2.95(m,1H),2.65-2.59(m,1H),1.97-1.86(m,2H),1.82-1.76(m,3H),1.47-1.42(m,2H),1.27-1.20(m,1H). 13 C NMR(151MHz,DMSO-d6)δ157.06,143.58,143.02,138.35,131.21,126.19,113.79,105.93,103.17,77.17,76.90, 73.79,70.44,66.80,61.45,55.67,52.00,51.33,45.19,44.50,29.60,26.81,24.35,23.18.HRMS(ESI):m / z[M+H] + Calcd for C 24 H 35 N4O7:491.2506; Found:491.2508.

[0198] Preparation of (23) (2R,3S,4S,5R,6R)-2-(hydroxymethyl)-6-(3-(9-methoxy-1,2,3,4,4a,5-hexahydro-6H-pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)tetrahydro-2H-pyran-3,4,5-triol (IV-20)

[0199]

[0200] The preparation method is the same as in "Example 6-(4)". A yellow solid (IV-20) was obtained by column chromatography, 130 mg, yield 21.8%, melting point 69.8-72.4℃. 1 H NMR (600MHz, DMSO-d6) δ7.31(d,J=8.8Hz,1H),6.87(d,J=2.8Hz,1H),6.79(dd,J=8.8,2.8Hz,1H),4.86(dd,J=12.0,4.6Hz ,1H),4.70(d,J=5.4Hz,1H),4.61-4.56(m,1H),4.56-4.53(m,1H),4.34(dd,J=4.6,2.5Hz,1H),4.11(dd,J=7.4,3.8Hz,1H ),3.85v3.81(m,1H),3.78(s,3H),3.74-3.62(m,3H),3.61-3.56(m,1H),3.56-3.50(m,2H),3.50-3.45(m,1H),3.37-3.35 (m,1H),3.34-3.26(m,4H),2.64-2.58(m,1H),1.97-1.86(m,2H),1.83-1.75(m,3H),1.49-1.40(m,2H),1.27-1.20(m,1H). 13 C NMR(151MHz,DMSO-d6)δ157.08,143.57,142.98,138.45,131.27,126.22,113.81,105.98,103.81,75.64,73.94, 71.10,68.60,66.67,60.91,55.67,52.04,51.42,45.23,44.45,29.56,26.95,24.41,23.23.HRMS(ESI):m / z[M+H] + Calcd for C 24 H 35 N4O7:491.2506; Found:491.2505.

[0201] (24) Preparation of (2R,3R,4S,5R,6R)-2-(2-(9-chloro-1,2,3,4,4a,5-6H-hexahydro[1',2':1,6]pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)-6-(hydroxymethyl)tetrahydro-2Hpyran-3,4,5-triol (IV-21)

[0202]

[0203] The preparation method is the same as in "Example 6-(4)". The white solid (IV-21) was obtained by column chromatography, 110 mg, yield 29.7%, melting point 130.7-106.6℃. 1 H NMR (600MHz, DMSO-d6) δ7.38(d,J=8.6Hz,1H),7.36(d,J=2.4Hz,1H),7.15(dd,J=8.6,2.4Hz,1H),5.03(dd,J=5.1,3.8Hz,1H),4.93(dd,J= 4.9,3.9Hz,1H),4.89(dd,J=5.3,1.2Hz,1H),4.67-4.62(m,1H),4.48-4.42(m,1H),4.19(dd,J=7.8,1.3Hz,1H),4.03-3.96(m,1H),3.94-3. 86(m,1H),3.83-3.76(m,1H),3.76-3.73(m,1H),3.73-3.68(m,1H),3 .67-3.63(m,1H),3.46-3.39(m,2H),3.39-3.36(m,1H),3.15-3.11(m ,1H),3.11-3.07(m,1H),3.07-3.01(m,1H),3.00-2.93(m,1H),2.71- 2.65(m,1H),1.85-1.75(m,3H),1.50-1.40(m,2H),1.28-1.21(m,1H). 13 C NMR(151MHz,DMSO-d6)δ144.44,143.68,137.86,135.35,128.40,126.57,123.94,123.42,103.27,76.89, 76.65,73.67,70.30,66.49,61.30,51.95,51.70,47.94,44.46,29.48,24.23,22.94.HRMS(ESI):m / z[M+H] + Calcd for C 22 H 30 N4O6Cl:481.1854; Found:481.1856.

[0204] Preparation of (25) (2R,3S,4S,5R,6R)-2-(2-(9-chloro-1,2,3,4,4a,5-hexahydro-6H-hexahydro[1',2':1,6]pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)ethoxy)-6-(hydroxymethyl)tetrahydro-2Hpyran-3,4,5-triol (IV-22)

[0205]

[0206] The preparation method is the same as in "Example 6-(4)". A pale yellow solid (IV-22) was obtained by column chromatography, 120 mg, yield 27.3%, melting point 202.2-205.0℃. 1 H NMR (600MHz, DMSO-d6) δ7.38(d,J=8.7Hz,1H),7.36(d,J=2.4Hz,1H),7.14(dd,J=8.6,2.5Hz,1H),4.90-4.85(m,1 H),4.70(dd,J=5.4,2.1Hz,1H),4.67-4.62(m,1H),4.54-4.51(m,1H),4.36-4.33(m,1H),4.13(d,J=7.3Hz,1H),4 .01-3.95(m,1H),3.91-3.84(m,1H),3.80-3.66(m,3H),3.63-3.61(m,1H),3.53-3.40(m,3H),3.40-3.35(m,1H), 3.33-3.27(m,2H),3.27-3.24(m,1H),2.70-2.64(m,1H),1.83-1.76(m,3H),1.50-1.38(m,2H),1.28-1.21(m,1H). 13 C NMR(151MHz,DMSO-d6)δ144.45,143.66,137.96,135.42,128.34,126.63,123.87,123.47,103.97,75.40, 73.54,70.88,68.42,66.32,60.71,51.94,51.77,48.06,44.43,29.48,24.27,22.99.HRMS(ESI):m / z[M+H] + Calcd forC 22 H 29 N4O6ClNa:503.1673; Found:503.1670.

[0207] (26) Preparation of (2R,3R,4S,5R,6R)-2-(3-(9-chloro-1,2,3,4,4a,5-6H-hexahydro[1',2':1,6]pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)-6-(hydroxymethyl)tetrahydro-2Hpyran-3,4,5-triol (IV-23)

[0208]

[0209] The preparation method is the same as in "Example 6-(4)". A pale yellow solid (IV-23) was obtained by column chromatography, 100 mg, yield 13.0%, melting point 118.8-121.5℃. 1 H NMR (600MHz, DMSO-d6) δ7.37(d,J=3.9Hz,1H),7.36(d,J=2.2Hz,1H),,7.13(dd,J=8.6,2.4Hz,1H),5.01(dd,J=13.7,4.9Hz,1H), 4.93(dd,J=4.9,1.2Hz,1H),4.89(d,J=5.2Hz,1H),4.67-4.63(m,1H),4.46(t,J=5.9Hz,1H),4.15(dd,J=7.8,3.9Hz,1H),3.88-3. 82(m,1H),3.73-3.60(m,4H),3.55-3.50(m,1H),3.46-3.42(m,1H),3.42-3.35(m,2H),3.17-3.13(m,1H),3.11-3.08(m,1H),3.0 7-3.02(m,1H),3.00-2.95(m,1H),2.71-2.65(m,1H),1.98-1.85(m,2H),1.84-1.75(m,3H),1.51-1.40(m,2H),1.29-1.22(m,1H). 13 C NMR(151MHz,DMSO-d6)δ144.38,143.77,138.09,135.36,128.32,126.58,123.80,123.48,103.05,76.97,76.76, 73.76,61.36,66.62,61.36,51.94,50.89,45.36,44.46,39.02,29.58,26.66,24.24,23.00.HRMS(ESI):m / z[M+H] + Calcd for C 23 H 32 N4O6Cl:495.2010; Found:495.2012.

[0210] (27) Preparation of (2R,3S,4S,5R,6R)-2-(3-(9-chloro-1,2,3,4,4a,5-6H-hexahydro[1',2':1,6]pyrido[1',2':1,6]pyrazino[2,3-b]quinoxalin-6-yl)propoxy)-6-(hydroxymethyl)tetrahydro-2Hpyran-3,4,5-triol (IV-24)

[0211]

[0212] The preparation method is the same as in "Example 6-(4)". The white solid (IV-24) was obtained by column chromatography, 80 mg, yield 11.9%, melting point 83.5-86.3℃. 1 H NMR (600MHz, DMSO-d6) δ7.38-7.37(m,1H),7.37-7.36(m,1H),7.13(dd,J=8.6,2.4Hz,1H),4.85(dd,J=13 .0,4.6Hz,1H),4.69(d,J=5.4Hz,1H),4.67-4.63(m,1H),4.55-4.52(m,1H),4.35-4.33(m,1H),4.10(dd, J=7.4,4.4Hz,1H),3.85-3.80(m,1H),3.75-3.60(m,4H),3.56-3.45(m,3H),3.42-3.35(m,2H),3.33-3.2 6(m,3H),2.71-2.65(m,1H),1.97-1.86(m,2H),1.84-1.76(m,3H),1.51-1.40(m,2H),1.29-1.21(m,1H). 13 CNMR(151MHz,DMSO-d6)δ144.39,143.79,138.29,135.51,128.26,126.66,123.70,123.65,103.80,75.66,73. 93,71.10,68.60,66.29,60.90,51.94,51.11,45.41,,45.39,29.61,26.86,24.36,23.12.HRMS(ESI):m / z[M+H] + Calcd for C 23 H 32 N4O6Cl:495.2010; Found:495.2015.

[0213] Thirdly, the following specific experimental examples demonstrate the beneficial effects of the present invention;

[0214] Experimental Example 1: Detection of the antitumor activity of the anticancer drug TNBG-glycoconjugate by CCK-8 assay

[0215] The antitumor activity of TNBG-glycoconjugates (Formulas I and IV) against the human lung adenocarcinoma cell line A549 was tested using the CCK-8 assay, with TNBG and sorafenib as positive controls. A549 cells were seeded in DMEM medium containing 10% FBS (fetal bovine serum) and cultured in a cell incubator at 37°C and 5% CO2. Cells were centrifuged, resuspended during the logarithmic growth phase, and then seeded in 96-well plates. After complete cell adhesion and growth, the specified concentration of TNBG-glycoconjugate was added to the wells and incubated for 72 h. The original medium was discarded, and 10% CCK-8 working solution was added. After incubation for 40 min, the OD value was measured at 450 nm. Data analysis was performed using SPSS 21.0 software. Inhibition rate was used as the response frequency, and drug concentration was used as the dependent variable. The concentration was transformed using Logit, and the total number of observations was set to 100. Probit regression analysis was then performed to calculate the IC50. 50 The final test result is expressed as IC value. 50 (Inhibition concentration 50%) is expressed as "mean ± standard deviation". The antitumor activity of the anticancer drug TNBG-glycoconjugate of this invention against A549 cells is shown in Table 1.

[0216] Table 1. Antitumor activity of the anticancer drug TNBG-glycoconjugate against A549 cell line.

[0217]

[0218] Note: "a" represents the average of three repeated trials.

[0219] As shown in Table 1, the anticancer drugs TNBG-glycoconjugates IV-12, IV-14 and IV-15 all exhibited good antitumor activity and have good application prospects.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, Formula I in, n is an integer selected from 1 to 6; G is selected from monosaccharides, wherein the monosaccharides are selected from glucose, galactose, and mannose; R1-R8 are selected from hydrogen, halogen, C1-C8 alkyl, C1-C8 alkoxy, hydroxyl, nitro, amino, and carboxyl.

2. The compound of formula I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, n is selected from the integer 3, and G is selected from D-glucose and D-galactose.

3. The compound represented by Formula II or a pharmaceutically acceptable salt thereof, Formula II in, n is an integer selected from 1 to 6; G is selected from monosaccharides, wherein the monosaccharides are selected from glucose, galactose, and mannose; R1-R4 are selected from hydrogen, halogen, C1-C8 alkyl, C1-C8 alkoxy, hydroxyl, nitro, amino, and carboxyl.

4. The compound of formula II according to claim 3, or a pharmaceutically acceptable salt thereof, wherein, n is selected from integers 2 and 3; R 1-4 The alkyl group is selected from hydrogen, the alkyl group is selected from methyl, the halogen is selected from F and Cl, the alkoxy group is selected from methoxy, and G is selected from D-glucose and D-galactose.

5. The compound according to claims 1 and 3, wherein the compound is selected from the following compounds: I-1 I-2 Ⅱ-1 Ⅱ-2 Ⅱ-3 Ⅱ-4 Ⅱ-5 Ⅱ-6 Ⅱ-7 Ⅱ-8 Ⅱ-9 Ⅱ-10 Ⅱ-11 Ⅱ-12 Ⅱ-13 Ⅱ-14 Ⅱ-15 Ⅱ-16 Ⅱ-17 Ⅱ-18 Ⅱ-19 Ⅱ-20 Ⅱ-21 Ⅱ-22 Ⅱ-23 Ⅱ-24。 6. Use of the compound of claim 5 or a pharmaceutically acceptable salt thereof in the preparation of a medicament beneficial for the treatment and / or prevention of cancer.

7. The use according to claim 6, characterized in that: The cancers mentioned are lung cancer and liver cancer.

8. A medicament comprising a formulation prepared by means of the compound of claims 1 to 5 or a pharmaceutically acceptable salt thereof as an active ingredient, and in combination with suitable excipients or auxiliary ingredients.

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