CN113024617A

CN113024617A - N-aryl sulfonamide-N-beta-D-glucopyranose diamide compounds and application thereof

Info

Publication number: CN113024617A
Application number: CN202110280125.4A
Authority: CN
Inventors: 刘洋; 程卯生; 张作鹏; 仲烨
Original assignee: Shenyang Pharmaceutical University
Current assignee: Shenyang Pharmaceutical University
Priority date: 2021-03-16
Filing date: 2021-03-16
Publication date: 2021-06-25
Anticipated expiration: 2041-03-16
Also published as: CN113024617B

Abstract

The invention belongs to the technical field of medicines, and relates to a compound with N¹arylsulfonamide-N⁴A preparation method and medical application of the-beta-D-glucopyranose diamide compound. The compound is shown as a general formula (I), the details of substituent groups are described in the specification, and the compound of the general formula (I) and an optically active body, diastereoisomers and pharmaceutically acceptable salts thereof are applied to preparation of antitumor drugs. The invention designs and synthesizes selective CA IX and XII inhibitors based on the pharmacophore characteristics and subcellular localization of CA IX and XII, selects polyhydroxy high-polarity glucose as a tail end, and passes through the softThe sex fat chain and the rigid aromatic structure introduce a targeting CAS active center classical drug effect group aryl sulfonamide segment, the whole structure can selectively inhibit extracellular CA IX and XII catalytic activity, and the anti-tumor effect is exerted, so that the application prospect is good.

Description

N-aryl sulfonamide-N-beta-D-glucopyranose diamide compounds and application thereof

Technical Field

The inventionBelongs to the technical field of medicine, and relates to a compound with N¹arylsulfonamide-N⁴A preparation method and medical application of the-beta-D-glucopyranose diamide compound.

Background

Carbonic Anhydrases (CAs) are a class of zinc-containing metalloenzymes widely found in living organisms, and their main function is to catalyze CO in living organisms₂And further participate in a series of related physiological activities. Currently, there are 15 subtypes of CAs found in humans, of which carbonic anhydrases IX (CA IX) and XII (CA XII) are hardly expressed in normal tissues or cells, but are highly expressed in various solid tumors, both of which have been identified as tumor-associated proteins due to CA IX and XII involved in microenvironment acidification of tumor tissues, tumor cell invasion and migration (m.y.mboge, et al.top Anti cancer Res,2015,5: 3; a.nocentini, et al.expert Opin Ther Pat,2018,28: 729-. The inhibitor is taken as an anti-tumor drug action target, and the research of the inhibitor is developed into a novel anti-tumor strategy which is concerned.

CA IX and XII are transmembrane proteins, the catalytic domain outside the cell membrane is the key region for the two to exert activity, the catalytic domain of each subtype of CAs has high similarity, and the structural characteristics are about one

Deep conical cavity with a zinc ion (Zn) at the bottom²⁺) With three histidines (His 94, His 96, His 119) and one water molecule/hydroxide ion to form a tetrahedral structure. The high degree of homology and essentially identical biological function of the various subtypes of CAs has contributed greatly to the design of highly selective CA IX and XII inhibitors (m.a. said, et al. eur J Med Chem,2020,185: 111843).

Due to the high similarity of the structures of various CAs in humans, it is extremely difficult to design, develop selective CA IX or CA XII inhibitors based on protein structures. In order to solve the problem, scientists propose a 'non-membrane permeability' theory of 'tail end modification', and according to different CAs subcellular localization, the aim of targeting different CAs subtypes is achieved by carrying out structural modification on the tail end of a pharmacophore. A variety of candidate CAS inhibitory active compounds with high single-tail, double-tail and even triple-tail activity and good selectivity have been obtained by this theoretical approach (M.A.Abdelrahman, et al.Eur J Med Chem,2021,209: 112897; A.Bonardi, et al.J Med Chem,2020,63: 7422-. By applying a non-membrane permeation theory, the reasonable improvement of the polarity of the compound and the reduction of the cell membrane permeability become important means for developing targeted extracellular CA subtype inhibitors. The inhibitor has the characteristics of less influence on the activity of cytoplasmic subtypes, effective reduction of side effects caused by drug off-target and improvement of the drug property of the inhibitor, so that the inhibitor is widely used and favored in the research and development of CAs inhibitors.

Disclosure of Invention

The invention aims to design and synthesize a novel N-containing compound¹arylsulfonamide-N⁴-beta-D-glucopyranose diamide compounds, for pharmaceutical formulation studies with efficacy in the treatment of cancer.

To achieve the purpose of the invention, the following technical scheme can be adopted:

an N-aryl sulfonamide-N-beta-D-glucopyranose diamide compound shown as a general formula (I)

In the formula (I), the compound is shown in the specification,

wherein:

n is an integer of 0-5, R is H, halogen, cyano, hydroxyl, NH₂、NO₂C1-C6 alkyl, C1-C6 alkoxy, C1-C6 halogenated alkyl or benzyl, m is an integer of 0-3, Linker is aryl or carboxylic acid derivative, and ZBG is aryl sulfonamide;

or, the compound shown in the general formula (I) and the optical active body, diastereoisomer and pharmaceutically acceptable salt thereof.

Preferably, N is represented by the general formula (I)¹arylsulfonamide-N⁴In the-beta-D-glucopyranose diamide compound, n is 1-4, R is H, halogen, CN, OH, NH₂Benzyl, CH₃、C₂H₅、C₃H₇、OCH₃、OC₂H₅、OC₃H₇、CF₃Or NO₂M is an integer of 1-3, Linker is aryl, amide or ester group, ZBG is arylsulfonamide; or, the compound shown in the general formula (I) and the optical active body, diastereoisomer and pharmaceutically acceptable salt thereof.

More preferably, in the general formula (I), n is 2 or 3, R is H, halogen, CN, OH, NH₂、Bn、CH₃、C₂H₅、C₃H₇、OCH₃、OC2H₅、OC₃H₇、CF₃Or NO₂M is 1 or 2, Linker is 1,2, 3-triazole, ester or amide, ZBG is benzenesulfonamide or thiazole sulfonamide; or, the compound shown in the general formula (I) and the optical active body, diastereoisomer and pharmaceutically acceptable salt thereof.

More preferably, in the general formula (I), n is 2 or 3, R is H, F, Cl, Br, CH₃、OCH₃2,6-2F, 2,6-2Cl, 2,6-2Br or NO₂The Linker is 1,2, 3-triazole or amide, and the ZBG is benzene sulfonamide or thiazole sulfonamide; or, the compound shown in the general formula (I) and the optical active body, diastereoisomer and pharmaceutically acceptable salt thereof.

More preferably, the compound is any of A1-A16 and B1-B15

A preparation method of N-aryl sulfonamide-N-beta-D-glucopyranose diamide compounds comprises the following steps:

performing benzoyl protection on glucose to obtain fully benzoylated glucose, performing terminal group bromination and terminal group azide substitution on the fully benzoylated glucose to obtain terminal group azide substituted benzoylated glucose, and reacting the azide substituted benzoylated glucose with corresponding anhydride under an alkaline condition to prepare a sugar donor containing a fatty chain;

when the Linker is a carboxylic acid derivative, after Fmoc protection is carried out on various substituted aminobenzoic acids, generating corresponding acyl chloride under the action of thionyl chloride, condensing with 5-amino-1, 3, 4-thiadiazole-2-sulfonamide obtained by removing acetyl from acetazolamide to obtain a corresponding thiadiazole sulfonamide structure, condensing with the obtained sugar donor in the presence of EDCI after Fmoc protection of thiadiazole sulfonamide is removed, and finally removing benzoyl protection under an alkaline condition to obtain a compound with a general formula (I) of which the Linker is the carboxylic acid derivative;

when the Linker is an aryl group, the Linker is,

diazotizing sulfanilamide to obtain azidobenzene sulfonamide, performing acetylation protection, Sonogashira coupling reaction and deprotection on H various substituted bromoaniline to obtain an ethynyl substituted corresponding structure, preparing a corresponding triazole sulfonamide structure by Click reaction between an alkynyl substituted compound and the azidobenzene sulfonamide under the action of a catalyst, condensing the triazole sulfonamide structure with the obtained sugar donor under the action of EDCI after removing acetyl protection, and finally removing protection under an alkaline condition to obtain the compound with the general formula (I) in which a Linker is an aryl group.

The application of the compound of the general formula (I) and an optically active body, diastereoisomer and pharmaceutically acceptable salt thereof in preparing antitumor drugs.

An anti-tumor composition comprises an active ingredient and a pharmaceutically acceptable carrier; wherein the active ingredient contains the compound of the general formula (I) and an optically active body, diastereoisomer and pharmaceutically acceptable salt thereof.

The present invention has limitations:

the invention designs and synthesizes selective CA IX and XII inhibitors based on the pharmacophore characteristics of CA IX and XII and subcellular localization, selects polyhydroxy high-polarity glucose as the tail end, introduces a classical pharmacophore arylsulfonamide segment of a targeting CAs active center through a flexible fatty chain and a rigid aromatic structure, and can selectively inhibit the catalytic activity of extracellular CA IX and XII by the whole structure to play an anti-tumor role; meanwhile, in the preparation of the intermediate, the one-pot method is adopted, azido reduction and ring opening reaction are completed by terminal group azido sugar at one time, and the preparation of important sugar donor fragments is completed.

Detailed Description

The compounds of the present invention and their preparation will be better understood in connection with the following examples, which are intended to illustrate, but not to limit, the scope of the invention.

Example 1: n is a radical of¹- (4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁴-beta-D-glucopyranose succinamide (A1)

Dissolving glucose (9.0g and 50.0mmol) in 50mL pyridine, adding a catalytic amount of DMAP, slowly dropwise adding benzoyl chloride (34.6mL and 300.0mmol) in ice bath, removing the ice bath after dropwise adding, reacting at room temperature for 5h, adding 200mL water, stirring for 10min, extracting with dichloromethane (3X100mL), washing an organic layer with water (2X100mL), adjusting the pH to subacidity with dilute hydrochloric acid, adjusting the pH to neutrality with saturated sodium bicarbonate solution, and drying over night with anhydrous sodium sulfate. Filtration and removal of the solvent under reduced pressure gave intermediate 2.

Dissolving the crude product in a proper amount of dichloromethane, dropwise adding 25mL of HBr-HOAc solution under an ice bath, removing the ice bath after the dropwise adding is finished, reacting for 4h at room temperature, adding 50mL of water, stirring for 10min, extracting with dichloromethane (3X100mL), washing an organic layer with water (2X100mL), adjusting the pH value to be neutral by using a saturated sodium bicarbonate solution, and drying over night by using anhydrous sodium sulfate. Column chromatography (petroleum ether: ethyl acetate: 5:1(v/v)) gave 22.0g of intermediate 3 in 66.9% yield over the two steps.

Intermediate 3(10.0g,15.2mmol) was weighed and dissolved in an appropriate amount of N, N-dimethylformamide, and NaN was added₃(4.9g,76.0mmol), heating to 50 ℃ for 6h, cooling to room temperature, adding 200mL of water, stirring for 10min, extracting with dichloromethane (2X100mL), washing with saturated sodium chloride solution, and drying over night over anhydrous sodium sulfate. Column chromatography (petroleum ether: ethyl acetate: 6:1(v/v)) afforded 7.2g of intermediate 4, in 76.3% yield.

Dissolving intermediate 4(3.6g,5.8mmol) in 30mL dichloromethane, adding catalytic amount of DMAP and palladium on carbon, adding succinic anhydride (1.2g,11.6mmol) under ice bath, removing ice bath, introducing hydrogen to react at room temperature for 8h, filtering off palladium on carbon, washing the filtrate with 10mL diluted hydrochloric acid, and drying over night with anhydrous sodium sulfate. Column chromatography (dichloromethane: methanol ═ 50:1) afforded 3.3g of succinic anhydride intermediate 5, in 81.8% yield.

Dissolving p-aminobenzoic acid (1.0g,7.3mmol) and sodium bicarbonate (0.9g,10.7mmol) in a mixed solution (20mL) of tetrahydrofuran and water (1: 1), slowly adding a tetrahydrofuran solution of FmocCl (2.8g,10.8mmol) dropwise, reacting at 50 ℃ for 5 hours after the dropwise addition, cooling to room temperature, concentrating the reaction solution, adding ethyl acetate (80mL), separating to obtain an organic layer, washing the organic layer with a saturated sodium chloride solution, and drying over night with anhydrous sodium sulfate. Filtering, decompressing and removing the solvent to obtain a crude intermediate 7.

The crude product was distributed in 20mL of SOCl₂And carrying out reflux reaction at 80 ℃ for 5h, stopping heating after the reaction solution becomes clear, standing and cooling to room temperature, and concentrating to remove the solvent to obtain 2.0g of a crude intermediate 8.

The preparation method comprises the steps of distributing acetazolamide (10.0g,45.0mmol) in 150mL of ethanol, adding 12mL of concentrated hydrochloric acid, performing reflux reaction for 10 hours, cooling to room temperature after reaction liquid becomes clear, concentrating the reaction liquid, adding saturated sodium bicarbonate to dissolve and adjust the pH value to be neutral, performing suction filtration, and collecting filter cakes to obtain 6.9g of intermediate 10, wherein the yield is 85.2%.

Dissolving the intermediate 10(1.8g,10.0mmol) in acetone, adding pyridine (1.9mL,23.2mmol), slowly adding the acetone solution of the intermediate 8 dropwise in ice bath, reacting at room temperature overnight, filtering, and collecting the filter cake to obtain 3.0g of the intermediate 11 with a yield of 57.6%.

Intermediate 11(3.0g,5.8mmol) was dissolved in 15mL of N, N-dimethylformamide, 10mL of diethylamine (97.1mmol) was added, reacted at room temperature for 3h, 50mL of water was added, extracted with ethyl acetate (2X50mL), the organic phases were combined, washed with saturated sodium chloride solution, and dried over anhydrous sodium sulfate overnight. Column chromatography (dichloromethane: methanol ═ 40:1) afforded 0.8g of intermediate 12, yield 46.5%.

Intermediate 5(0.5g,0.7mmol), intermediate 12(0.3g,1.0mmol) and EDCI (0.2g, 1.0mmol) were dissolved in 15mL pyridine, reacted at 60 ℃ for 48h, cooled to room temperature and the reaction was concentrated to give intermediate 13.

The intermediate 13 was dissolved in acetone, and 0.5mL of 1N sodium methoxide in methanol was added thereto under ice-cooling, followed by reaction at room temperature for 2 hours. Cation exchange resin was added to adjust pH to neutral, filtration was performed, the filtrate was concentrated, and column chromatography was performed (dichloromethane: methanol 9:1) to obtain 40.0mg of a white solid, with a yield of 10.2% in two steps.

¹H NMR(600MHz,DMSO-d₆)δ13.36(s,1H),10.39(s,1H),8.44(d,J＝9.1Hz,1H),8.35(s,2H),8.13(d,J＝8.7Hz,2H),7.78(d,J＝8.7Hz,2H),4.87(s,1H),4.71(t,J＝9.1Hz,1H),3.67–3.59(m,1H),3.41(dd,J＝11.8,5.1Hz,1H),3.35(s,1H),3.17(t,J＝8.6Hz,1H),3.11–3.01(m,3H),2.65–2.59(m,2H),2.56–2.51(m,2H).ESI-MS(m/z):559.0[M-H]^-。

Example 2: n is a radical of¹- (3- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁴-beta-D-glucopyranose succinamide (A2)

The compound of example 2 was prepared in the same manner as in example 1 except that m-aminobenzoic acid was used instead of p-aminobenzoic acid, to obtain 45.1mg of a white solid with a yield of 11.5%.

¹H NMR(600MHz,DMSO-d₆)δ13.55(s,1H),10.24(s,1H),8.44(d,J＝9.1Hz,1H),8.35(d,J＝18.7Hz,3H),7.94–7.74(m,2H),7.49(t,J＝7.9Hz,1H),4.98(d,J＝4.5Hz,1H),4.87(dd,J＝11.3,5.2Hz,2H),4.71(t,J＝9.1Hz,1H),4.51(t,J＝5.7Hz,1H),3.63(dd,J＝11.5,4.0Hz,1H),3.48–3.37(m,1H),3.17(td,J＝8.6,4.4Hz,1H),3.12–2.99(m,3H),2.66–2.56(m,2H),2.50–2.44(m,2H).ESI-MS(m/z):559.3[M-H]^-。

Example 3: n is a radical of¹- (2-fluoro-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁴-beta-D-glucopyranose succinamide (A3)

The compound of example 3 was prepared in the same manner as in example 1 except that 3-fluoro-4-aminobenzoic acid was used instead of p-aminobenzoic acid, and 26.1mg of a white solid was obtained with a yield of 6.4%.

¹H NMR(600MHz,DMSO-d₆)δ9.95(s,1H),8.43(d,J＝9.1Hz,1H),8.12(t,J＝7.9Hz,1H),7.99–7.88(m,4H),4.97(s,1H),4.86(dd,J＝9.3,5.2Hz,3H),4.75–4.65(m,2H),4.51(t,J＝6.0Hz,2H),3.63(dd,J＝10.3,5.0Hz,2H),3.45–3.38(m,2H),3.21–3.13(m,2H),3.11–2.99(m,5H),2.72–2.62(m,2H),2.46–2.32(m,2H).ESI-MS(m/z):577.2[M-H]^-。

Example 4: n is a radical of¹- (2-chloro-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁴-beta-D-glucopyranose succinamide (A4)

The compound of example 4 was prepared as in example 1 except that 3-chloro-4-aminobenzoic acid was used instead of p-aminobenzoic acid to give 35.4mg of a white solid with a yield of 8.5%.

¹H NMR(600MHz,DMSO-d₆)δ13.26(s,1H),8.42(d,J＝9.0Hz,1H),8.14(d,J＝2.0Hz,1H),7.88(dd,J＝8.6,2.0Hz,1H),6.87(d,J＝8.6Hz,1H),6.43(s,2H),4.65(t,J＝9.0Hz,1H),3.60(d,J＝11.3Hz,1H),3.40(dd,J＝11.8,4.5Hz,1H),3.25–3.10(m,1H),3.03(dd,J＝16.3,7.9Hz,3H),2.61–2.53(m,2H),2.44–2.27(m,2H).ESI-MS(m/z):593.3[M-H]^-。

Example 5: n is a radical of¹- (2-bromo-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁴-beta-D-glucopyranose succinamide (A5)

The compound of example 5 was prepared as in example 1 except that 3-bromo-4-aminobenzoic acid was used instead of p-aminobenzoic acid to give 22.3mg of a white solid with a yield of 5.0%.

¹H NMR(600MHz,DMSO-d₆)δ13.25(s,1H),8.39(d,J＝9.1Hz,1H),8.29(d,J＝2.0Hz,1H),7.90(dd,J＝8.6,2.0Hz,1H),6.85(d,J＝8.6Hz,1H),6.37(s,2H),4.66(t,J＝9.1Hz,1H),3.61(d,J＝10.5Hz,1H),3.39(dd,J＝11.8,4.9Hz,2H),3.15(dd,J＝15.2,6.6Hz,1H),3.10–2.95(m,3H),2.56–2.51(m,2H),2.35(td,J＝16.8,9.6Hz,2H).ESI-MS(m/z):637.5[M-H]^-。

Example 6: n is a radical of¹- (2-methyl-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁴-beta-D-glucopyranose succinamide (A6)

The compound of example 6 was prepared in the same manner as in example 1 except that 3-methyl-4-aminobenzoic acid was used instead of p-aminobenzoic acid, to give 33.5mg of a white solid with a yield of 8.3%.

¹H-NMR(600MHz,DMSO-d₆)δ13.36(s,1H),9.50(s,1H),8.45(d,J＝9.0Hz,1H),8.36(s,1H),8.04(s,1H),7.91(dd,J＝83.8,8.5Hz,2H),4.72(t,J＝9.0Hz,1H),3.64(d,J＝11.0Hz,1H),3.42(dd,J＝11.7,5.1Hz,1H),3.22–3.15(m,3H),3.12–3.02(m,4H),2.75(dd,J＝11.7,4.4Hz,2H),2.68–2.61(m,2H),2.33(s,3H).ESI-MS(m/z):573.1[M-H]^-。

Example 7: n is a radical of¹- (2-methoxy-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁴-beta-D-glucopyranose succinamide (A7)

The compound of example 7 was prepared in the same manner as in example 1 except that 3-methoxy-4-aminobenzoic acid was used instead of p-aminobenzoic acid, to give 40.5mg of a white solid with a yield of 9.8%.

¹H NMR(600MHz,DMSO-d₆)δ13.43(s,1H),9.48(s,1H),8.42(d,J＝9.1Hz,1H),8.36(s,2H),8.30(d,J＝8.3Hz,1H),7.85(s,1H),7.78(d,J＝9.9Hz,1H),4.85(s,1H),4.71(t,J＝9.1Hz,1H),3.97(s,3H),3.63(d,J＝10.5Hz,1H),3.60(s,1H),3.41(dd,J＝11.8,5.1Hz,2H),3.17(t,J＝8.7Hz,1H),3.12–3.01(m,3H),2.77(t,J＝6.6Hz,1H),2.75–2.64(m,2H),2.49–2.41(m,2H).ESI-MS(m/z):589.0[M-H]^-。

Example 8: n is a radical of¹- (2-nitro-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁴-beta-D-glucopyranose succinamide (A8)

The compound of example 8 was prepared in the same manner as in example 1 except that 3-nitro-4-aminobenzoic acid was used instead of p-aminobenzoic acid, to give 30.2mg of a white solid with a yield of 7.1%.

¹H NMR(600MHz,DMSO-d₆)δ13.64(s,1H),9.00(d,J＝2.1Hz,1H),8.41(d,J＝9.0Hz,1H),8.09(dd,J＝8.9,2.2Hz,3H),7.13(d,J＝9.0Hz,1H),4.65(t,J＝9.0Hz,1H),3.60(d,J＝10.5Hz,2H),3.39(dd,J＝11.8,4.7Hz,2H),3.21–3.11(m,1H),3.11–2.97(m,3H),2.58–2.51(m,3H),2.43–2.28(m,2H).ESI-MS(m/z):604.1[M-H]^-。

Example 9: n is a radical of¹- (4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (A9)

The compound of example 9 was prepared as in example 1 except that glutaric anhydride was used instead of succinic anhydride to give 46.7mg of a white solid in 11.6% yield.

¹H NMR(600MHz,DMSO-d₆)δ10.05(s,1H),8.35(d,J＝9.1Hz,1H),8.24(s,1H),8.07(d,J＝8.7Hz,2H),7.74(s,2H),7.64(d,J＝8.6Hz,2H),4.72(t,J＝9.1Hz,1H),3.64(dd,J＝11.7,1.8Hz,1H),3.41(dd,J＝11.8,5.4Hz,1H),3.17(t,J＝8.8Hz,1H),3.12–2.99(m,3H),2.36(dd,J＝13.9,6.6Hz,2H),2.18(ddq,J＝22.5,15.0,7.5Hz,2H),1.93–1.72(m,2H).ESI-MS(m/z):573.5[M-H]^-。

Example 10: n is a radical of¹- (3- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (A10)

The compound of example 10 was prepared in the same manner as in example 9 except that m-aminobenzoic acid was used instead of p-aminobenzoic acid to obtain 40.3mg of a white solid with a yield of 10.0%.

¹H NMR(600MHz,DMSO-d₆)δ13.48(s,1H),10.12(s,1H),8.35(d,J＝2.0Hz,2H),8.19(s,2H),7.83(d,J＝7.9Hz,2H),7.45(t,J＝7.9Hz,1H),4.96(d,J＝4.1Hz,1H),4.91–4.81(m,2H),4.72(t,J＝9.1Hz,1H),4.49(t,J＝5.1Hz,1H),3.64(dd,J＝11.0,3.9Hz,1H),3.44–3.38(m,1H),3.17(td,J＝8.7,3.5Hz,1H),3.11–3.01(m,3H),2.37(t,J＝7.4Hz,2H),2.27–2.07(m,2H),1.88–1.78(m,2H).ESI-MS(m/z):573.0[M-H]^-。

Example 11: n is a radical of¹- (2-fluoro-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (A11)

The compound of example 11 was prepared in the same manner as in example 9 except that 3-fluoro-4-aminobenzoic acid was used instead of p-aminobenzoic acid to give 40.1mg of a white solid with a yield of 9.7%.

¹H NMR(600MHz,DMSO-d₆)δ9.77(s,1H),8.36(d,J＝9.1Hz,1H),8.25(s,1H),8.05–7.97(m,1H),7.92(d,J＝8.4Hz,1H),7.86(d,J＝12.1Hz,1H),7.70(s,2H),4.71(dt,J＝13.4,9.1Hz,2H),3.63(d,J＝14.1Hz,2H),3.44–3.37(m,2H),3.22–3.13(m,2H),3.12–3.00(m,4H),2.44(t,J＝7.1Hz,2H),2.24–2.16(m,2H),1.82(dt,J＝14.7,7.3Hz,2H).ESI-MS(m/z):591.3[M-H]^-。

Example 12: n is a radical of¹- (2-chloro-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (A12)

The compound of example 12 was prepared in the same manner as in example 9 except that 3-chloro-4-aminobenzoic acid was used instead of p-aminobenzoic acid to give 33.7mg of a white solid with a yield of 7.9%.

¹H NMR(600MHz,DMSO-d₆)δ12.58(s,1H),8.23(d,J＝9.1Hz,1H),8.10(s,1H),7.85(dd,J＝8.6,1.8Hz,1H),6.84(d,J＝8.6Hz,1H),6.29(s,2H),4.88(d,J＝41.0Hz,3H),4.70(t,J＝9.1Hz,1H),4.54(s,1H),3.62(d,J＝11.7Hz,1H),3.40(d,J＝11.4Hz,3H),3.16(t,J＝8.7Hz,1H),3.11–2.99(m,4H),2.07(dq,J＝14.6,7.4Hz,2H),2.00(t,J＝7.3Hz,2H),1.63(p,J＝7.3Hz,2H).ESI-MS(m/z):607.5[M-H]^-。

Example 13: n is a radical of¹- (2-bromo-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (A13)

The compound of example 13 was prepared as in example 9 except that 3-bromo-4-aminobenzoic acid was used instead of p-aminobenzoic acid to give 24.1mg of a white solid with a yield of 5.3%.

¹H-NMR(600MHz,DMSO-d₆)δ13.23(s,1H),8.35(d,J＝8.9Hz,1H),8.26(d,J＝1.8Hz,1H),7.92(dd,J＝8.2,1.8Hz,1H),6.79(d,J＝8.2Hz,1H),6.35(s,2H),4.65(t,J＝8.5Hz,1H),3.53(d,J＝9.1Hz,1H),3.32(dd,J＝11.0,4.4Hz,2H),3.18(dd,J＝13.5,6.1Hz,1H),3.10–2.95(m,3H),2.17–1.95(m,4H),1.68(d,J＝35.5Hz,2H).ESI-MS(m/z):651.0[M-H]^-。

Example 14: n is a radical of¹- (2-methyl-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (A14)

The compound of example 14 was prepared in the same manner as in example 9 except that 3-methyl-4-aminobenzoic acid was used instead of p-aminobenzoic acid to give 45.8mg of a white solid with a yield of 11.1%.

¹H-NMR(600MHz,DMSO-d₆)δ13.21(s,1H),9.37(s,1H),8.35(d,J＝9.1Hz,1H),8.15(s,2H),8.03(d,J＝1.3Hz,1H),7.96(dd,J＝8.4,1.6Hz,1H),7.70(dd,J＝23.7,8.3Hz,1H),4.97(d,J＝0.5Hz,1H),4.87(dd,J＝11.1,4.8Hz,2H),4.73(t,J＝9.1Hz,1H),4.49(t,J＝5.1Hz,1H),3.64(dd,J＝11.5,2.4Hz,1H),3.45–3.38(m,1H),3.18(t,J＝8.4Hz,1H),3.13–2.98(m,3H),2.43(t,J＝7.1Hz,2H),2.30(s,3H),2.27–2.11(m,2H),1.84(p,J＝7.3Hz,2H).ESI-MS(m/z):588.7[M-H]^-。

Example 15: n is a radical of¹- (2-methoxy-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (A15)

The compound of example 15 was prepared as in example 9 except that 3-methoxy-4-aminobenzoic acid was used instead of p-aminobenzoic acid to give 40.4mg of a white solid with a yield of 9.6%.

¹H NMR(600MHz,DMSO-d₆)δ13.44(s,1H),9.38(s,1H),8.36(s,2H),8.34(d,J＝9.1Hz,1H),8.30(d,J＝8.4Hz,1H),7.85(d,J＝1.8Hz,1H),7.78(dd,J＝8.5,1.8Hz,1H),4.90(s,1H),4.72(t,J＝9.1Hz,1H),3.97(s,3H),3.64(dd,J＝11.7,1.7Hz,1H),3.41(dd,J＝11.8,5.4Hz,1H),3.18(t,J＝8.8Hz,1H),3.14–2.95(m,3H),2.54–2.51(m,2H),2.29–2.01(m,2H),1.91–1.71(m,2H).ESI-MS(m/z):603.1[M-H]^-。

Example 16: n is a radical of¹- (2-nitro-4- ((5-sulfamoyl-1, 3, 4-thiadiazol-2-yl) carbamoyl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (A16)

The compound of example 15 was prepared in the same manner as in example 9 except that 3-nitro-4-aminobenzoic acid was used instead of p-aminobenzoic acid to give 24.6mg of a white solid with a yield of 5.7%.

¹H NMR(600MHz,DMSO-d₆)δ13.55(d,J＝81.3Hz,1H),8.99(dd,J＝6.6,2.1Hz,1H),8.42–8.29(m,2H),8.10(d,J＝9.0Hz,3H),7.14(d,J＝9.0Hz,1H),4.87(s,1H),4.67(t,J＝9.0Hz,1H),3.65–3.57(m,1H),3.40(dd,J＝11.8,4.9Hz,2H),3.16(t,J＝8.6Hz,1H),3.11–2.99(m,2H),2.35(t,J＝7.1Hz,2H),2.10(ddt,J＝30.3,15.2,7.5Hz,2H),1.76–1.60(m,2H).ESI-MS(m/z):619.0[M-H]^-。

Example 17: n is a radical of¹- (4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁴-beta-D-glucopyranose succinamide (B1)

Dispersing sulfanilamide (1.0g,5.8mmol) in appropriate amount of water, adding concentrated hydrochloric acid (2mL) under ice bath condition, stirring until all is dissolved, adding sodium nitrite (0.5g,7.2mmol), stirring at room temperature for 2h, and preparing intermediate 15. Without separation, sodium azide (0.8g,12.3mmol) was added to the solution, reaction was continued for 0.5h, then dichloromethane (50mL) was added, the organic layer was washed successively with water, saturated sodium bicarbonate solution and saturated sodium chloride solution, dried over anhydrous sodium sulfate overnight, filtered, and the solvent was removed under reduced pressure to give crude intermediate 16.

P-bromoaniline (1.0g,5.8mmol) was dissolved in acetic acid (10mL), acetic anhydride (0.7mL,7.5mmol) was added, reaction was carried out at room temperature for 1.5 hours, dichloromethane (30mL) was then added, the organic layer was washed with water, a saturated sodium bicarbonate solution and a saturated sodium chloride solution in this order, dried over anhydrous sodium sulfate overnight, filtered, and the solvent was removed under reduced pressure to give 1.2g of intermediate 18, yield 97.1%.

After dissolving intermediate 18(1.2g,5.6mmol), bis (triphenylphosphine) palladium dichloride (0.2g,0.3mmol) and cuprous iodide (0.1g,0.5mmol) in an appropriate amount of N, N-dimethylformamide, triethylamine (7.2mL,51.6mmol) and trimethylethynyl silicon (1.2mL,8.7mmol) were added, reacted at 50 ℃ under argon atmosphere for 12h, ethyl acetate (50mL) was added, the organic layer was extracted with water and saturated sodium chloride solution in that order, dried over anhydrous sodium sulfate overnight, filtered, the solvent was removed under reduced pressure, and column chromatography (petroleum ether: ethyl acetate ═ 5:1) gave 1.0g of intermediate 19, 77.3% yield.

Intermediate 19(1.0g,4.3mmol) was dissolved in an appropriate amount of N, N-dimethylformamide, 1N tetrabutylammonium fluoride solution 2.0mL was added, reaction was carried out at room temperature for 3 hours, then ethyl acetate (30mL) was added, the organic layer was extracted with water and saturated sodium chloride solution in this order, dried over anhydrous sodium sulfate overnight, filtered, and the solvent was removed under reduced pressure to give 0.5g of intermediate 20 with a yield of 75.2%.

Dissolving the intermediate 16(0.6g,3.1mmol) and 20(0.5g,3.1mmol) in a tert-butanol-water (20mL,1:1) mixed solution, adding copper sulfate pentahydrate (75.0mg,0.3mmol) and sodium ascorbate (59.4mg,0.3mmol), reacting at 50 ℃ for 3h, cooling to room temperature, and concentrating the reaction solution to obtain an intermediate 21.

The crude intermediate 21 was distributed in 20mL of aqueous sodium hydroxide (2N), refluxed for 5 hours, cooled to room temperature, filtered, the filter cake was dissolved, and column chromatography was performed (dichloromethane: methanol: 25: 1) to obtain 0.5g of intermediate 22, with a yield of 51.2% in two steps.

Intermediate 5(0.5g,0.7mmol), intermediate 22(0.3g,1.0mmol) and EDCI (0.2g, 1.0mmol) were dissolved in 15mL pyridine, reacted at 60 ℃ for 48h, cooled to room temperature and the reaction was concentrated to give intermediate 23.

Intermediate 23 was dissolved in acetone, and 1N sodium methoxide in 0.5mL of methanol was added thereto under ice-cooling, followed by reaction at room temperature for 2 hours. Cation exchange resin was added to adjust pH to neutral, filtration was performed, the filtrate was concentrated, and column chromatography was performed (dichloromethane: methanol ═ 9:1) to obtain 45.0mg of a white solid, with a yield of 11.2% in two steps.

¹H NMR(600MHz,DMSO-d₆)δ10.16(s,1H),9.33(s,1H),8.51(d,J＝9.0Hz,1H),8.17(d,J＝8.7Hz,2H),8.06(d,J＝8.7Hz,2H),7.88(d,J＝8.6Hz,2H),7.73(d,J＝8.6Hz,2H),7.54(s,2H),5.10–4.90(m,2H),4.71(t,J＝9.0Hz,1H),4.53(s,1H),3.64(d,J＝10.7Hz,1H),3.45–3.40(m,1H),3.18(dd,J＝11.5,5.5Hz,1H),3.12–3.01(m,3H),2.63–2.56(m,2H),2.47(dd,J＝15.9,7.7Hz,1H).ESI-MS(m/z):575.6[M-H]^-

Example 18: n is a radical of¹- (3- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁴-beta-D-glucopyranose succinamide (B2)

The compound of example 18 was prepared as in example 17 except that m-bromoaniline was used instead of p-bromoaniline, and 40.1mg of a white solid was obtained in 9.9% yield.

¹H NMR(600MHz,DMSO-d₆)δ10.14(s,1H),9.37(s,1H),8.49(d,J＝8.9Hz,1H),8.32(s,1H),8.21(d,J＝8.7Hz,2H),8.05(d,J＝8.7Hz,2H),7.56(dd,J＝11.0,9.7Hz,4H),7.42(t,J＝7.9Hz,1H),4.95(d,J＝22.5Hz,3H),4.72(t,J＝9.0Hz,1H),4.50(s,1H),3.64(d,J＝11.5Hz,1H),3.42(d,J＝10.3Hz,1H),3.22–3.14(m,1H),3.07(q,J＝9.4Hz,3H),2.69–2.56(m,2H).ESI-MS(m/z):575.4[M-H]^-。

Example 19: n is a radical of¹- (2-fluoro-4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁴-beta-D-glucopyranose succinamide (B3)

The compound of example 19 was prepared as in example 17 except 2-fluoro-4-bromoaniline was used instead of para-bromoaniline to give 37.2mg of a white solid in 8.9% yield.

¹H NMR(600MHz,DMSO-d₆)δ9.93(s,1H),9.44(s,1H),8.54(d,J＝8.9Hz,1H),8.16(d,J＝8.6Hz,2H),8.07(d,J＝8.6Hz,3H),7.79(d,J＝11.8Hz,1H),7.74(d,J＝8.4Hz,1H),7.56(s,2H),5.07(s,2H),4.98(s,1H),4.71(t,J＝9.0Hz,1H),4.54(s,1H),3.63(d,J＝11.2Hz,1H),3.42(d,J＝10.8Hz,1H),3.18(t,J＝8.5Hz,1H),3.13–3.01(m,3H),2.72–2.58(m,2H),2.49–2.40(m,2H).ESI-MS(m/z):594.3[M-H]^-。

Example 20: n is a radical of¹- (2-chloro-4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁴-beta-D-glucopyranose succinamide (B4)

The compound of example 20 was prepared as in example 17 except that 2-chloro-4-bromoaniline was used instead of para-bromoaniline, and 30.0mg of a white solid was obtained in 7.0% yield.

¹H NMR(600MHz,DMSO-d₆)δ12.29(s,1H),9.26(s,1H),8.36(d,J＝9.1Hz,1H),8.15(dd,J＝24.2,8.8Hz,4H),7.77(d,J＝1.8Hz,1H),7.62(dd,J＝8.3,1.8Hz,1H),6.90(d,J＝8.4Hz,1H),5.64(s,2H),4.95(d,J＝4.3Hz,1H),4.84(dd,J＝19.3,5.1Hz,2H),4.65(t,J＝9.1Hz,1H),4.49(t,J＝5.6Hz,1H),3.60(dd,J＝11.7,4.0Hz,1H),3.42–3.36(m,1H),3.14(td,J＝8.7,4.0Hz,1H),3.08–2.96(m,3H),2.47–2.39(m,2H),2.37–2.23(m,2H).ESI-MS(m/z):609.1[M-H]^-。

Example 21: n is a radical of¹- (2, 6-difluoro-4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl)) Phenyl) -N⁴-beta-D-glucopyranose succinamide (B5)

The compound of example 21 was prepared as in example 17 except 2, 6-difluoro-4-bromoaniline was used instead of para-bromoaniline to give 20.2mg of a white solid in 4.7% yield.

¹H NMR(600MHz,DMSO-d₆)δ9.25(d,J＝43.4Hz),8.39–8.17(m),8.02(ddd,J＝36.2,32.2,8.6Hz),7.54(s),7.48(d,J＝7.4Hz),5.49(d,J＝17.4Hz),4.96(s),4.85(s),4.72–4.61(m),4.52(s),3.67–3.56(m),3.43–3.37(m),3.16(dd,J＝17.5,8.7Hz),3.05(dd,J＝16.8,7.4Hz),2.45–2.17(m).ESI-MS(m/z):611.3[M-H]^-。

Example 22: n is a radical of¹- (2-methyl-4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁴-beta-D-glucopyranose succinamide (B6)

The compound of example 22 was prepared as in example 17 except that 2-methyl-4-bromoaniline was used instead of p-bromoaniline, and 29.6mg of a white solid was obtained in 7.2% yield.

¹H NMR(600MHz,DMSO-d₆)δ9.39(d,J＝18.3Hz,2H),8.51(d,J＝9.0Hz,1H),8.18(d,J＝8.7Hz,2H),8.06(d,J＝8.7Hz,2H),7.80(s,1H),7.73(d,J＝9.3Hz,1H),7.61(d,J＝8.2Hz,1H),7.55(s,3H),4.74(s,1H),3.64(d,J＝10.6Hz,1H),3.42(dd,J＝11.8,5.2Hz,2H),3.19(t,J＝8.7Hz,1H),3.14–2.99(m,3H),2.67–2.57(m,2H),2.30(s,4H).ESI-MS(m/z):589.4[M-H]^-。

Example 23: n is a radical of¹- (2-methoxy-4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁴-beta-D-glucopyranose succinamide (B7)

The compound of example 23 was prepared as in example 17 except 2-methoxy-4-bromoaniline was used instead of para-bromoaniline to give 37.8mg of a white solid in 8.9% yield.

¹H NMR(600MHz,DMSO-d₆)δ9.41(s,1H),9.29(s,1H),8.59(d,J＝8.8Hz,1H),8.17(d,J＝8.6Hz,2H),8.14(d,J＝8.2Hz,1H),8.07(d,J＝8.6Hz,2H),7.60(s,1H),7.56(s,2H),7.50(d,J＝9.5Hz,1H),5.18(s,2H),5.04(s,1H),4.70(t,J＝9.0Hz,1H),4.56(s,1H),3.95(s,3H),3.63(d,J＝11.0Hz,1H),3.42(d,J＝11.5Hz,1H),3.18(t,J＝8.4Hz,1H),3.10–3.01(m,3H),2.66(dd,J＝25.0,17.2Hz,2H),2.46(dd,J＝16.1,8.2Hz,2H).ESI-MS(m/z):605.5[M-H]^-。

Example 24: n is a radical of¹- (4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (B8)

The compound of example 24 was prepared as in example 17 except that glutaric anhydride was used instead of succinic anhydride to give 23.9mg of a white solid in 5.8% yield.

¹H NMR(600MHz,DMSO-d₆)δ10.13(s),9.36(s),8.18(d,J＝8.2Hz),8.06(d,J＝8.3Hz),7.88(d,J＝7.9Hz),7.75(d,J＝7.6Hz),7.55(s),5.01(s),4.90(dd,J＝12.8,4.3Hz),4.72(t,J＝8.5Hz),4.51(s),3.64(dd,J＝10.6,4.3Hz),3.42(d,J＝6.1Hz),3.18(s),3.07(dd,J＝9.7,4.7Hz),3.03–2.93(m),2.64–2.53(m),2.41–2.33(m),2.20(ddd,J＝27.9,14.0,6.9Hz),1.89–1.78(m).ESI-MS(m/z):589.4[M-H]^-。

Example 25: n is a radical of¹- (3- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (B9)

The compound of example 25 was prepared as in example 24 except that m-bromoaniline was used instead of p-bromoaniline to give 29.0mg of a white solid in 7.0% yield.

¹H NMR(600MHz,DMSO-d₆)δ10.06(s,1H),9.38(s,1H),8.50–8.25(m,2H),8.21(d,J＝8.5Hz,2H),8.06(d,J＝8.5Hz,2H),7.58(dd,J＝18.9,11.5Hz,4H),7.43(t,J＝7.8Hz,1H),4.98(s,1H),4.88(s,2H),4.73(t,J＝9.0Hz,1H),4.48(s,1H),3.64(d,J＝8.6Hz,1H),3.48–3.38(m,1H),3.19(t,J＝8.4Hz,1H),3.13–2.97(m,3H),2.38(t,J＝7.1Hz,2H),2.20(dd,J＝14.2,7.1Hz,2H),1.96–1.74(m,2H).ESI-MS(m/z):589.0[M-H]^-。

Example 26: n is a radical of¹- (2-fluoro-4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (B10)

The compound of example 26 was prepared as in example 24 except that 2-fluoro-4-bromoaniline was used instead of para-bromoaniline, and 32.0mg of a white solid was obtained in 7.5% yield.

¹H NMR(600MHz,DMSO-d₆)δ9.87(d,J＝51.7Hz),9.44(s),8.16(d,J＝8.7Hz),8.07(d,J＝8.6Hz),7.79(d,J＝13.2Hz),7.75(d,J＝8.2Hz),7.55(s),4.98(d,J＝3.9Hz),4.91–4.83(m),4.73(td,J＝9.1,5.4Hz),4.50(dt,J＝11.3,5.8Hz),3.64(dd,J＝11.4,5.4Hz),3.48–3.39(m),3.18(td,J＝8.7,4.3Hz),3.14–3.01(m),2.71–2.59(m),2.44(t,J＝7.3Hz),2.19(ddt,J＝22.5,15.1,7.5Hz),1.89–1.77(m).ESI-MS(m/z):607.8[M-H]^-。

Example 27: n is a radical of¹- (2-chloro-4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (B11)

The compound of example 27 was prepared as in example 24 except 2-chloro-4-bromoaniline was used in place of para-bromoaniline to give 29.1mg of a white solid in 6.7% yield.

¹H NMR(600MHz,DMSO-d₆)δ12.21(s,1H),9.25(s,1H),8.26(d,J＝9.1Hz,1H),8.16(q,J＝8.9Hz,4H),7.77(d,J＝1.8Hz,1H),7.62(dd,J＝8.4,1.8Hz,1H),6.90(d,J＝8.4Hz,1H),5.62(s,2H),4.92(s,1H),4.81(d,J＝15.1Hz,2H),4.68(t,J＝9.0Hz,1H),4.44(s,1H),3.62(d,J＝11.3Hz,1H),3.40(dd,J＝11.5,4.2Hz,1H),3.23–3.12(m,2H),3.05(dt,J＝17.8,9.2Hz,3H),2.28(t,J＝7.3Hz,2H),2.07(d,J＝8.0Hz,2H),1.73–1.53(m,2H).ESI-MS(m/z):623.1[M-H]^-。

Example 28: n is a radical of¹- (2, 6-difluoro-4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (B12)

The compound of example 28 was prepared as in example 24, except 2, 6-difluoro-4-bromoaniline was used instead of para-bromoaniline, and 20.9mg of a white solid was obtained in 4.8% yield.

¹H NMR(600MHz,DMSO-d₆)δ12.25(s),9.31(s),8.28(d,J＝9.1Hz),8.21–8.03(m),7.49(dd,J＝7.4,2.1Hz),5.51(s),4.95(d,J＝4.4Hz),4.86(d,J＝5.2Hz),4.82(d,J＝5.0Hz),4.68(t,J＝9.1Hz),4.47(t,J＝5.4Hz),3.62(dd,J＝11.4,3.2Hz),3.45–3.36(m),3.15(td,J＝8.7,4.1Hz),3.11–3.05(m),3.02(td,J＝9.1,5.1Hz),2.27(t,J＝7.4Hz),2.16–1.97(m),1.63(p,J＝7.4Hz).ESI-MS(m/z):624.9[M-H]^-。

Example 29: n is a radical of¹- (2-methyl-4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (B13)

The compound of example 29 was prepared as in example 24 except 2-methyl-4-bromoaniline was used instead of para-bromoaniline to give 39.9mg of a white solid in 9.4% yield.

¹H-NMR(400MHz,DMSO-d₆)δ9.31(s),8.17(d,J＝8.7Hz),8.07(d,J＝8.7Hz),7.81(s),7.74(d,J＝9.6Hz),7.58(d,J＝8.2Hz),4.74(d,J＝9.0Hz),3.65(d,J＝11.5Hz),3.42(dd,J＝11.8,5.5Hz),3.21(t,J＝8.8Hz),3.14(dd,J＝8.3,6.6Hz),3.07(dd,J＝19.1,10.1Hz),2.41(t,J＝7.0Hz),2.30(s),2.23(dd,J＝12.3,7.2Hz),1.92–1.80(m).ESI-MS(m/z):603.5[M-H]^-。

Example 30: n is a radical of¹- (2-methoxy-4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (B14)

The compound of example 30 was prepared as in example 24 except that 2-methoxy-4-bromoaniline was used instead of p-bromoaniline, and 47.1mg of a white solid was obtained in 10.8% yield.

¹H NMR(600MHz,DMSO-d₆)δ9.41(s,1H),9.18(s,1H),8.37(t,J＝24.4Hz,1H),8.17(d,J＝8.8Hz,2H),8.13(d,J＝8.2Hz,1H),8.07(d,J＝8.8Hz,2H),7.60(d,J＝1.7Hz,1H),7.55(s,2H),7.51(dd,J＝8.2,1.7Hz,1H),4.98(d,J＝4.4Hz,1H),4.88(dd,J＝10.1,5.4Hz,2H),4.73(t,J＝9.1Hz,1H),4.50(t,J＝5.8Hz,1H),3.95(s,3H),3.68–3.60(m,1H),3.41(dt,J＝11.5,5.7Hz,1H),3.18(td,J＝8.7,3.8Hz,1H),3.14–2.99(m,3H),2.44(t,J＝7.1Hz,2H),2.26–2.08(m,2H),1.82(p,J＝7.5Hz,2H).ESI-MS(m/z):619.6[M-H]^-。

Example 31: n is a radical of¹- (2-nitro-4- (1- (4-sulfamoylphenyl) -1H-1,2, 3-triazol-4-yl) phenyl) -N⁵-beta-D-glucopyranose glutaramide (B14)

The compound of example 31 was prepared as in example 24 except 2-nitro-4-bromoaniline was used instead of para-bromoaniline to give 19.9mg of a white solid in 4.5% yield.

¹H-NMR(400MHz,DMSO-d₆)δ12.32(s,1H),9.38(s,1H),8.55(d,J＝1.7Hz,1H),8.24(d,J＝9.0Hz,1H),8.08(q,J＝8.6Hz,4H),7.98(dd,J＝8.8,1.7Hz,1H),7.64(s,2H),7.17(d,J＝8.8Hz,1H),4.92(d,J＝3.1Hz,1H),4.83(d,J＝3.8Hz,2H),4.68(t,J＝9.0Hz,1H),4.46(s,1H),3.62(d,J＝11.4Hz,1H),3.39(dd,J＝12.2,3.5Hz,2H),3.15(d,J＝12.8Hz,2H),3.10–2.93(m,4H),2.16(s,2H),2.06(dd,J＝13.1,7.1Hz,2H),1.70–1.51(m,2H).ESI-MS(m/z):634.2[M-H]^-。

Pharmacological study of the products of the invention

(1) The inhibitory effect of each of the compounds A1-16 and B1-15 on human carbonic anhydrases XI and XII (hCA IX and XII) was determined by enzymatic hydrolysis. The substrate 4-nitrophenylacetate was diluted to 2mM with assay buffer, the enzyme was diluted to 20 ng/. mu.L with formulated assay buffer (125mM Tris,75mM NaCl), 50. mu.L of 20 ng/. mu.L enzyme was added to a 96-well plate, each of the compounds prepared above was added as an inhibitor, incubated at room temperature for 15 minutes, 25. mu.L of 2mM substrate was added to initiate the reaction, and the absorbance value at 405nm was read by a microplate reader after 10 minutes. Testing the inhibition rate of 8 different concentrations to obtain IC₅₀The value, inhibition ratio calculation formula is as follows:

the results obtained for% inhibition (1-sample activity/total activity) X100 are given in the following table:

n.t.not test.

(2) the cell viability inhibitory effects of preferred compounds A7, A9, A10, A14, B1, B2 and B6 on human colon cancer cells HT-29 were determined using the MTT method. Taking the above cells in logarithmic growth phase, adjusting cell density to 2X10⁴Embedding the wells in 96-well plate, culturing at 37 deg.C for 24 hr, setting three parallel wells in each group except negative control group, adding corresponding medicine, and keeping at 37 deg.CAfter 24 hours of incubation, MTT (0.5mg/mL) solution was added to each well for 3 hours, the supernatant was discarded, after washing the residual drug with PBS, 100. mu. LDMSO was added to each well, and the mixture was shaken in a micro shaker for 10 minutes until the crystals were completely dissolved, followed by measuring the absorbance at 540nM of each well with an Emax Microplate Reader (Molecular Devices, Sunnyvale, Calif., USA), and the cell viability inhibition was calculated from the negative control:

Inhibitory ratio(％)＝(A_540,control-A_{540,experiment})/(A_540,control-A_540,blank) X100 the test results obtained are given in the following table:

the pharmacological research shows that the compound of the invention shows obvious inhibition activity to the tumor-related protein CA IX in the pharmacological test of the enzyme level in vitro, and the half inhibition concentration obtained by another tumor-related protein CA XII activity inhibition experiment carried out on the partial compound is 41-256 nM; subsequent tumor cell viability inhibition experiments with the preferred compounds tested for enzyme levels showed that the compounds inhibited human colon cancer cells HT-29 by 22.2-43.1%.

In conclusion, according to the pharmacophore characteristics and subcellular distribution characteristics of tumor-related proteins CA IX and CA XII, terminal group azido sugar is prepared through multi-step reactions, and then azido is reduced through a one-pot method and subjected to ring-opening reaction to synthesize a sugar donor, so that the preparation of the N-arylsulfonamide-N-beta-D-glucopyranose diamide compound is completed; enzyme level pharmacological activity tests show that the glucopyranose diamide compounds designed by the invention have good CA IX and XII inhibitory activity, wherein a compound with an amide structure in Linker shows better inhibitory activity than triazole derivatives, and halogen substitution can cause activity reduction, and cell experiment results show that the compounds designed by the invention show a certain inhibitory effect on human colon cancer cells HT-29 and have potential application prospects.

Claims

1. An N-aryl sulfonamide-N-beta-D-glucopyranose diamide compound is characterized in that: the compound is shown as the general formula (I)

In the formula (I), the compound is shown in the specification,

wherein:

2. The N-arylsulfonamide-N- β -D-glucopyranose diamide compound of claim 1, wherein: n is represented by the general formula (I)¹arylsulfonamide-N⁴In the-beta-D-glucopyranose diamide compound, n is 1-4, R is H, halogen, CN, OH, NH₂Benzyl, CH₃、C₂H₅、C₃H₇、OCH₃、OC₂H₅、OC₃H₇、CF₃Or NO₂M is an integer of 1-3, Linker is aryl, amide or ester group, ZBG is arylsulfonamide; or, the compound shown in the general formula (I) and the optical active body, diastereoisomer and pharmaceutically acceptable salt thereof.

3. The N-arylsulfonamide-N- β -D-glucopyranose diamide compound of claim 2, wherein: in the formula

n is 2 or 3, R is H, halogen, CN, OH, NH₂、Bn、CH₃、C₂H₅、C₃H₇、OCH₃、OC2H₅、OC₃H₇、CF₃Or NO₂M is 1 or 2, Linker is 1,2, 3-triazole, ester or amide,ZBG is benzenesulfonamide or thiazole sulfonamide; or, the compound shown in the general formula (I) and the optical active body, diastereoisomer and pharmaceutically acceptable salt thereof.

4. The N-arylsulfonamide-N- β -D-glucopyranose diamide compound of claim 3, wherein: in the formula, n is 2 or 3, R is H, F, Cl, Br, CH₃、OCH₃2,6-2F, 2,6-2Cl, 2,6-2Br or NO₂The Linker is 1,2, 3-triazole or amide, and the ZBG is benzene sulfonamide or thiazole sulfonamide; or, the compound shown in the general formula (I) and the optical active body, diastereoisomer and pharmaceutically acceptable salt thereof.

5. The N-arylsulfonamide-N- β -D-glucopyranose diamide compound of claim 4, wherein: the compound is any one of compounds A1-A16 and B1-B15

6. A method for preparing the N-arylsulfonamide-N- β -D-glucopyranose diamide compound of claim 1, characterized in that:

when the Linker is an aryl group, the Linker is,

7. Use of a compound according to claim 1, wherein: use of compounds of general formula (I) according to any of claims 1 to 5, as well as their optically active forms, diastereoisomers and pharmaceutically acceptable salts, for the preparation of antitumor medicaments.

8. An anti-tumor composition, comprising: the active ingredients of the composition and a pharmaceutically acceptable carrier; wherein the active ingredient comprises a compound of the general formula (I) as defined in any one of claims 1 to 5, and optically active forms, diastereomers and pharmaceutically acceptable salts thereof.