SULFONYLUREA DERIVATIVE AND MEDICAL USES THEREOF

Abstract

The present invention relates to a sulfonylurea derivative and medical uses thereof, specifically relating to the sulfonylurea derivative shown in general formula (I), a preparation method thereof, a pharmaceutical composition containing same, and a use of same for treating diseases and disorders affected by neuronal damage, for example: cerebral stroke, brain damage, neuropathic pain, migraines, inflammatory pain, chronic pain, or depression. The definition of each group in the general formula (I) is the same as that in the description.

##STR00001##

Claims

1. A compound of general formula (I) or a pharmaceutically acceptable salt or isomer thereof, ##STR00035## wherein, R.sub.1 are each independently selected from the group consisting of deuterium, alkyl optionally substituted with halogen, halogen, hydroxy, mercapto, —NR.sub.iR.sub.j, —C(O)R.sub.k, —C(O)OR.sub.k, nitro, cyano, alkoxy optionally substituted with halogen, and alkylthio; R.sub.2 is selected from the group consisting of alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl and heterocyclyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl and heterocyclyl are optionally further substituted with one or more groups selected from the group consisting of deuterium, alkyl, haloalkyl, halogen, hydroxy, oxo, mercapto, —NR.sub.iR.sub.j, —C(O)R.sub.k, —C(O)OR.sub.k, —SR.sub.i, —S(O)R.sub.i, —SO.sub.2R.sub.i, nitro, cyano, aryl optionally substituted with hydroxy or alkoxy, heteroaryl optionally substituted with oxo or alkyl, heterocyclyl optionally substituted with oxo or alkyl, cycloalkyl optionally substituted with oxo or alkyl, optionally substituted alkenyl, optionally substituted alkynyl, alkoxy, and alkylthio; R.sub.3 are each independently selected from the group consisting of deuterium, alkyl optionally substituted with halogen, halogen, hydroxy, mercapto, —NR.sub.iR.sub.j, —C(O)R.sub.k, —C(O)OR.sub.k, nitro, cyano, alkoxy, and alkylthio; R.sub.i and R.sub.j are each independently selected from the group consisting of hydrogen, hydroxy, alkyl, cycloalkyl, alkoxy, and —C(═NH)—NH.sub.2; R.sub.k is selected from the group consisting of alkyl, alkoxy, aryl, alkenyl and alkynyl, wherein the alkyl, alkoxy, aryl, alkenyl and alkynyl are optionally further substituted with one or more groups selected from the group consisting of aryl, heteroaryl, heterocyclyl, cycloalkyl, alkyl and alkoxy; m is an integer selected from the group consisting of 0 to 5; n is an integer selected from the group consisting of 0 to 10; and R.sub.2 is not ##STR00036##

2. The compound of general formula (I) or the pharmaceutically acceptable salt or isomer thereof according to claim 1, wherein R.sub.1 is selected from the group consisting of halogen, C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkyl optionally substituted with halogen, and cyano.

3. The compound of general formula (I) or the pharmaceutically acceptable salt or isomer thereof according to claim 1, wherein R.sub.1 is selected from the group consisting of Cl, OCH.sub.3, OCD.sub.3, CN, OH, and OCF.sub.3.

4. The compound of general formula (I) or the pharmaceutically acceptable salt or isomer thereof according to claim 1, wherein, R.sub.2 is selected from the group consisting of ##STR00037##

5. The compound of general formula (I) or the pharmaceutically acceptable salt or isomer thereof according to claim 1, wherein n is 0.

6. The compound of general formula (I) or the pharmaceutically acceptable salt or isomer thereof according to claim 1, wherein n is 0, and R.sub.1 is selected from the group consisting of halogen, C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkyl optionally substituted with halogen, and cyano.

7. The compound of general formula (I) or the pharmaceutically acceptable salt or isomer thereof according to claim 1, wherein n is 0, and R.sub.1 is selected from the group consisting of Cl, OCH.sub.3, CN, OH, and OCF.sub.3.

8. The compound of general formula (I) or the pharmaceutically acceptable salt or isomer thereof according to claim 1, wherein n is 0, and R.sub.2 is selected from the group consisting of ##STR00038##

9. The compound of general formula (I) or the pharmaceutically acceptable salt or isomer thereof according to claim 1, wherein, R.sub.1 is selected from the group consisting of halogen, C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkyl optionally substituted with halogen, and cyano, R.sub.2 is selected from the group consisting of ##STR00039## and n is 0.

10. The compound of general formula (I) or the pharmaceutically acceptable salt or isomer thereof according to claim 1, wherein, R.sub.1 is selected from the group consisting of Cl, OCH.sub.3, CN, OH and OCF.sub.3, R.sub.2 is selected from the group consisting of ##STR00040## and n is 0.

11. A compound of general formula (I-1) or a pharmaceutically acceptable salt or isomer thereof, ##STR00041## wherein, R.sub.2 is selected from the group consisting of alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl and heterocyclyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl and heterocyclyl are optionally further substituted with one or more groups selected from the group consisting of deuterium, alkyl, haloalkyl, halogen, hydroxy, oxo, mercapto, —NR.sub.iR.sub.j, —C(O)R.sub.k, —C(O)OR.sub.k, —SR.sub.i, —S(O)R.sub.i, —SO.sub.2R.sub.i, nitro, cyano, aryl optionally substituted with hydroxy and/or alkoxy, heteroaryl optionally substituted with oxo and/or alkyl, heterocyclyl optionally substituted with oxo and/or alkyl, cycloalkyl optionally substituted with oxo and/or alkyl, optionally substituted alkenyl, optionally substituted alkynyl, alkoxy, and alkylthio; R.sub.i and R.sub.j are each independently selected from the group consisting of hydrogen, hydroxy, alkyl, cycloalkyl, alkoxy, and —C(═NH)—NH.sub.2; R.sub.k is selected from the group consisting of alkyl, alkoxy, aryl, alkenyl and alkynyl, wherein the alkyl, alkoxy, aryl, alkenyl and alkynyl are optionally further substituted with one or more groups selected from the group consisting of aryl, heteroaryl, heterocyclyl, cycloalkyl, alkyl and alkoxy; and R.sub.2 is not ##STR00042##

12. The compound of general formula (I-1) or a pharmaceutically acceptable salt or isomer thereof according to claim 11, wherein R.sub.2 is selected from the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, and C.sub.6-C.sub.10 aryl; wherein the C.sub.1-C.sub.6 alkyl is optionally substituted with C.sub.6-C.sub.10 aryl or —NR.sub.iR.sub.j, wherein R.sub.i and R.sub.j are each independently selected from the group consisting of hydrogen and —C(═NH)—NH.sub.2; the C.sub.6-C.sub.10 aryl is optionally substituted with hydroxy and/or C.sub.1-C.sub.6 alkoxy, or the C.sub.6-C.sub.10 aryl is optionally substituted with 5- to 7-membered heterocyclyl, wherein the 5- to 7-membered heterocyclyl is optionally substituted with oxo and/or C.sub.1-C.sub.6 alkyl; and the C.sub.2-C.sub.6 alkenyl is optionally substituted with C.sub.6-C.sub.10 aryl, wherein the aryl is optionally substituted with hydroxy and/or C.sub.1-C.sub.6 alkoxy.

13. A compound as shown below or a pharmaceutically acceptable salt or isomer thereof, ##STR00043##

14. (canceled)

15. A pharmaceutical composition comprising the compound or the pharmaceutically acceptable salt or isomer thereof according to claim 1 and at least one pharmaceutically acceptable carrier, diluent or excipient.

16. A method for preventing or treating a disease or condition affected by neuronal damage in a subject in need thereof, the method comprising: administering to the subject the compound or the pharmaceutically acceptable salt or isomer thereof according to claim 1.

17. A method for treating acute stroke, traumatic brain injury, spinal cord injury, myocardial infarction, shock, organ ischemia, ventricular arrhythmia, ischemic injury, or hypoxia/ischemia in a subject in need thereof, the method comprising: administering to the subject the compound or the pharmaceutically acceptable salt or isomer thereof according to claim 1.

18. A method for preventing or treating a disease or condition affected by neuronal damage in a subject in need thereof, the method comprising: administering to the subject a compound as shown below or a pharmaceutically acceptable salt or isomer thereof, ##STR00044##

19. A method for preventing or treating a disease or condition affected by neuronal damage in a subject in need thereof, the method comprising: administering to the subject a compound as shown below or a pharmaceutically acceptable salt or isomer thereof, ##STR00045##

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0174] FIG. 1: concentration distribution of the compounds of the present disclosure in brain tissue 10 min after intravenous injection.

[0175] FIG. 2: concentration distribution of the compounds of the present disclosure in cerebrospinal fluid 10 min after intravenous injection.

[0176] FIG. 3: brain-to-plasma ratio of the compounds of the present disclosure 10 min after intravenous injection.

[0177] FIG. 4: cerebrospinal fluid-to-plasma ratio of the compounds of the present disclosure 10 min after intravenous injection.

DETAILED DESCRIPTION

[0178] The present disclosure is further described below with reference to examples, which are not intended to limit the scope of the present disclosure.

[0179] Experimental procedures without conditions specified in the examples of the present disclosure are generally conducted according to conventional conditions, or according to conditions recommended by the manufacturers of the starting materials or commercial products. Reagents without specific origins indicated are commercially available conventional reagents.

[0180] NMR shift (δ) is given in a unit of 10.sup.−6 (ppm). NMR spectra are determined using a Bruker AVANCE-400 nuclear magnetic resonance instrument, with deuterated dimethyl sulfoxide (DMSO-d.sup.6), deuterated chloroform (CDCl.sub.3) and deuterated methanol (CD.sub.3OD) as determination solvents, and tetramethylsilane (TMS) as an internal standard.

[0181] MS spectra are determined using a Shimadzu 2010 Mass Spectrometer or Agilent 6110A MSD Mass Spectrometer.

[0182] High-Performance Liquid Chromatography (HPLC) analysis is performed using Shimadzu LC-20A systems, Shimadzu LC-2010HT series, or Agilent 1200 LC high-performance liquid chromatograph (Ultimate XB-C18 3.0×150 mm chromatographic column or Xtimate C18 2.1×30 mm chromatographic column).

[0183] Chiral HPLC analysis is performed using the following chromatographic columns: Chiralpak IC-3 100×4.6 mm I.D., 3 μm, Chiralpak AD-3 150×4.6 mm I.D., 3 μm, Chiralpak AD-3 50×4.6 mm I.D., 3 μm, Chiralpak AS-3 150×4.6 mm I.D., 3 μm, Chiralpak AS-3 100×4.6 mm I.D., 3 μm, ChiralCel OD-3 150×4.6 mm I.D., 3 μm, Chiralcel OD-3 100×4.6 mm I.D., 3 μm, ChiralCel OJ-H 150×4.6 mm I.D., 5 μm, Chiralcel OJ-3 150×4.6 mm I.D., 3 μm.

[0184] Huanghai HSGF254 or Qingdao GF254 silica gel plates of specifications 0.15 mm to 0.2 mm are adopted for thin layer chromatography (TLC) analysis and 0.4 mm to 0.5 mm for TLC separation and purification.

[0185] Yantai Huanghai silica gel of 100-200 mesh, 200-300 mesh or 300-400 mesh is generally used as a carrier in column chromatography.

[0186] Chiral HPLC preparation is performed using a DAICEL CHIRALPAK IC (250×30 mm, 10 μm) or Phenomenex-Amylose-1 (250×30 mm, 5 μm) column.

A CombiFlash rapid preparation instrument used is Combiflash Rf150 (TELEDYNE ISCO).

[0187] The mean inhibition rate of kinase and the IC.sub.50 value are determined using a NovoStar microplate reader (BMG, Germany).

[0188] Known starting materials described herein may be synthesized using or according to methods known in the art, or may be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc., Chembee Chemicals, and other companies.

[0189] In the examples, the reactions can be performed in an argon atmosphere or a nitrogen atmosphere unless otherwise specified.

[0190] The argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to a balloon containing about 1 L of argon or nitrogen.

[0191] The hydrogen atmosphere means that the reaction flask is connected to a balloon containing about 1 L of hydrogen.

[0192] Parr 3916EKX hydrogenator, Qinglan QL-500 hydrogenator or HC2-SS hydrogenator was used in the pressurized hydrogenation reactions.

[0193] The hydrogenation reactions usually involve 3 cycles of vacuumization and hydrogen purge.

[0194] A CEM Discover-S 908860 microwave reactor is used in the microwave reactions.

[0195] In the examples, a solution refers to an aqueous solution unless otherwise specified.

[0196] In the examples, the reaction temperature is room temperature, i.e., 20° C. to 30° C., unless otherwise specified.

[0197] The monitoring of the reaction progress in the examples is conducted by thin layer chromatography (TLC). The developing solvent for reactions, the eluent system of column chromatography for compound purification and the developing solvent system of thin layer chromatography include: A: dichloromethane/methanol system, B: n-hexane/ethyl acetate system, C: petroleum ether/ethyl acetate system, and D: petroleum ether/ethyl acetate/methanol system. The volume ratio of the solvents is adjusted according to the polarity of the compound, or by adding a small amount of basic or acidic reagents such as triethylamine and acetic acid.

[0198] The abbreviations used in the following experiments have the following meanings: [0199] EtOAc: ethyl acetate; DCM: dichloromethane; DIPEA: N,N-diisopropylethylamine; PPTS: pyridinium p-toluenesulfonate; Boc: tert-butoxycarbonyl, MeOH: methanol.

Example 1: Preparation of 5-chloro-N-(4-(N-(cyclohexylcarbamoyl)sulfamoyl)phenethyl)-2-hydroxybenzamide

[0200] ##STR00028##

[0201] To a 50 mL reaction flask were added compound 1-1 (986 mg, 2 mmol, purchased from Cadila pharmaceuticals limited) and DCM (10 mL) under nitrogen atmosphere. The mixture was stirred and cooled to 0° C. under an ice bath, and a solution of BBr.sub.3 in DCM (3 mL, 3 mmol) was slowly added dropwise over a period of about 10 min. The mixture was stirred for 30 min while maintaining the ice bath. The ice bath was removed, and the mixture was naturally warmed to room temperature and stirred overnight. The reaction was quenched by dropwise addition of methanol (2 mL) under an ice bath. The reaction was concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography to give compound 1 (560 mg, 99.08% purity, 58.4% yield).

[0202] .sup.1HNMR (d.sup.6-DMSO, 400 MHz) δ1.06-1.30 (m, 5H), 1.47-1.65 (m, 5H), 2.96 (t, J=6.4 Hz, 2H), 3.32 (bs, 1H), 3.55-3.58 (m, 2H), 6.33 (d, J=7.6 Hz, 1H), 6.93 (d, J=8.8 Hz, 1H), 7.42-7.49 (m, 3H), 7.81-7.89 (m, 3H), 8.96 (s, 1H), 10.31 (bs, 1H), 12.46 (bs, 1H).

Example 2: Preparation of 5-chloro-N-(4-(N-(cyclohexylcarbamoyl)sulfamoyl)phenethyl)-2-(methoxy-d.SUB.3.)benzamide (2)

[0203] ##STR00029##

Step 1: preparation of methyl-d.SUB.3 .5-chloro-2-(methoxy-d.SUB.3.)benzoate (2-2)

[0204] To a 100 mL reaction flask were added 5-chloro-2-hydroxybenzoic acid (2-1) (5.18 g, 30.0 mmol), iodomethane (7.5 mL, 75.0 mmol), potassium carbonate (8.4 g, 61.0 mmol) and DMF (15 mL) at room temperature under nitrogen atmosphere. The mixture was heated to 60° C. and stirred for about 20 h. After the reaction was completed as monitored by LCMS, water (50 mL) and methyl tert-butyl ether (100 mL) were added to the reaction solution, and the aqueous phase was extracted with methyl tert-butyl ether (2×100 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was dissolved in toluene (50 mL) and washed with 5% sodium hydroxide solution (30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give crude product 2-2 (6.20 g), which was directly used in the next step.

Step 2: preparation of 5-chloro-2-(methoxy-d.SUB.3.)benzoic acid (2-3)

[0205] To a 100 mL flask were added compound 2-2 (6.20 g, 30 mmol), ethanol (30 mL) and 10% sodium hydroxide solution (30 mL) at room temperature. The mixture was heated to reflux and stirred for 2 h. After the reaction was substantially completed as monitored, the reaction solution was cooled to room temperature, and concentrated under reduced pressure to remove ethanol. To the resulting aqueous solution was added 5% dilute hydrochloric acid (60 mL) to form a suspension, which was then stirred for 1 h and filtered. The solid was dried in vacuum to give crude product 2-3 (5.5 g, 96.7% yield over two steps, 97% purity) as an off-white solid.

Step 3: preparation of 5-chloro-2-(methoxy-d.SUB.3.)-N-(4-sulfamoylphenethyl)benzamide (2-5)

[0206] To a 50 mL reaction flask were added compound 2-3 (3.79 g, 20.0 mmol), 4-(2-aminoethyl)benzenesulfonamide (2-4) (4.01 g, 20.0 mmol) and MeCN (15 mL) under nitrogen atmosphere. The mixture was stirred and cooled under an ice bath. Pyridine (2.5 mL, 40.0 mmol) and EDCI (3.84 g, 20.0 mmol) were added, and the mixture was stirred for 30 min while maintaining the ice bath. The ice bath was removed, and the mixture was naturally warmed to room temperature and stirred overnight. The reaction solution was concentrated under reduced pressure, and water (400 mL) was added at room temperature. The resulting mixture was stirred for 15 min. The reaction solution was filtered. The solid was collected and dried to give 2-5 (6.2 g, 99% purity, 83% yield) as an off-white solid, which was directly used in the next step without purification.

Step 4: preparation of 5-chloro-N-(4-(N-(cyclohexylcarbamoyl)sulfamoyl)phenethyl)-2-(methoxy-d.SUB.3.)benzamide (2)

[0207] Compound 2-5 (2.23 g, 6.0 mmol) and isocyanatocyclohexane (2-6) (1.28 g, 10.2 mmol) were dissolved in DMF (16 mL). The mixture was then cooled under an ice-water bath and a solution of potassium tert-butoxide (7.8 mL, 7.8 mmol, 1 M in THF) was slowly added dropwise over a period of about 5 min. After the addition, the mixture was warmed to room temperature and stirred overnight. Water (50 mL) was added and the suspension was stirred for an additional about 10 min. The resulting material was collected and purified by preparative HPLC to give compound 2 (1.9 g, 63.8% yield, 99.6% purity).

[0208] .sup.1HNMR (d.sup.6-DMSO, 400 MHz) δ 1.07-1.23 (m, 5H), 1.46-1.65 (m, 5H), 2.93 (t, J=6.4 Hz, 2H), 3.28 (bs, 1H), 3.54-3.57 (m, 2H), 6.33 (d, J=7.6 Hz, 1H), 7.14 (d, J=8.8 Hz, 1H), 7.45-7.64 (m, 4H), 7.84 (d, J=8.0 Hz, 2H), 8.27 (s, 1H), 10.31 (bs, 1H).

Example 3: Preparation of 5-chloro-N-(4-(N-(cyclohexylcarbamoyl)sulfamoyl)phenethyl)-2-(trifluoromethoxy)benzamide (3)

[0209] ##STR00030##

Step 1: preparation of 5-chloro-N-(4-sulfamoylphenethyl)-2-(trifluoromethoxy)benzamide (3-2)

[0210] To a 50 mL reaction flask were added 5-chloro-2-(trifluoromethoxy)benzoic acid (3-1) (288.7 g, 1.2 mmol), 4-(2-aminoethyl)benzenesulfonamide (2-4) (240.3 g, 1.2 mmol) and MeCN (2 mL) under nitrogen atmosphere. The mixture was stirred and cooled under an ice bath. Pyridine (0.2 mL, 2.4 mmol) and EDCI (230 mg, 1.2 mmol) were added, and the mixture was stirred for 30 min while maintaining the ice bath. The ice bath was removed, and the mixture was naturally warmed to room temperature and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography to give compound 3-2 (280 mg, 55% yield, 98.8% purity) as an off-white solid.

Step 2: preparation of 5-chloro-N-(4-(N-(cyclohexylcarbamoyl)sulfamoyl)phenethyl)-2-(trifluoromethoxy)benzamide (3)

[0211] Compound 3-2 (280 mg, 6.0 mmol) and isocyanatocyclohexane (2-6) (140 mg, 1.12 mmol) were dissolved in DMF (2.5 mL). The mixture was then cooled under an ice-water bath and a solution of potassium tert-butoxide (0.86 mL, 0.86 mmol, 1 M in THF) was slowly added dropwise over a period of about 5 min. After the addition, the mixture was warmed to room temperature and stirred overnight. Water (5 mL) was added and no solid was precipitated. The mixture was extracted with ethyl acetate (3×10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC to give compound 3 (240 mg, 66.4% yield, 99.69% purity).

[0212] .sup.1HNMR (d.sup.6-DMSO, 400 MHz) δ 1.05-1.25 (m, 5H), 1.46-1.66 (m, 5H), 2.91 (t, J=6.4 Hz, 2H), 3.32 (bs, 1H), 3.48-3.52 (m, 2H), 6.33 (d, J=7.6 Hz, 1H), 7.46-7.55 (m, 4H), 7.64 (d, J=8.8 Hz, 1H), 7.82 (d, J=8.0 Hz, 2H), 8.67 (s, 1H), 10.31 (bs, 1H).

Example 4: Preparation of 5-chloro-N-(4-(N-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)sulfamoyl)phenethyl)-2-methoxybenzamide (4)

[0213] ##STR00031##

[0214] (1r,4r)-4-aminocyclohexan-1-ol (4-2) (250 mg, 2.2 mmol) and ethyl ((4-(2-(5-chloro-2-methoxybenzamido)ethyl)phenyl)sulfonyl)carbamate (4-1) (880 mg, 2.0 mmol, synthesized according to the method described in the literature (Bioorganic & Medicinal Chemistry, 2003, 11, 2099-2113)) were added to toluene (30 mL), and the mixture was heated to reflux and stirred for about 3 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by preparative HPLC to give compound 4 (420 mg, 41.2% yield, 98.85% purity).

[0215] 1HNMR (d.sup.6-DMSO, 400 MHz) δ 1.02-1.24 (m, 4H), 1.66-1.76 (m, 4H), 2.08 (s, 1H), 2.94 (t, J=6.4 Hz, 2H), 3.22 (bs, 1H), 3.54-3.57 (m, 2H), 3.80 (s, 3H), 4.52 (bs, 1H), 6.30 (d, J=6.0 Hz, 1H), 7.15 (d, J=8.4 Hz, 1H), 7.47-7.51 (m, 3H), 7.65 (s, 1H), 7.85 (d, J=7.2 Hz, 2H), 8.28 (s, 1H), 10.35 (bs, 1H).

Example 5: Preparation of 5-chloro-N-(4-(N-((cyclohexyl-4,4-d.SUB.2.)carbamoyl)sulfamoyl)phenethyl)-2-methoxybenzamide (5)

[0216] ##STR00032##

[0217] Ethyl ((4-(2-(5-chloro-2-methoxybenzamido)ethyl)phenyl)sulfonyl)carbamate (4-1) (137 mg, 1.36 mmol) and cyclohexan-4,4-d.sub.2-1-amine (5-1) (500 mg, 1.13 mmol) were added to toluene (30 mL), and the mixture was heated to reflux and stirred for about 6 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by preparative HPLC to give compound 5 (380 mg, 67.9% yield, 100% HPLC purity).

[0218] .sup.1HNMR (d.sup.6-DMSO, 400 MHz) δ 1.04-1.23 (m, 4H), 1.54-1.66 (m, 4H), 2.93 (t, J=6.4 Hz, 2H), 3.28 (bs, 1H), 3.53-3.56 (m, 2H), 3.79 (s, 3H), 6.33 (d, J=6.4 Hz, 1H), 7.15 (d, J=8.8 Hz, 1H), 7.46-7.50 (m, 3H), 7.63 (s, 1H), 7.84 (d, J=7.6 Hz, 2H), 8.27 (bs, 1H), 10.31 (bs, 1H).

Example 6: Preparation of 5-chloro-N-(4-(N-(((1R,2R)-2-cyanocyclohexyl)carbamoyl)sulfamoyl)phenethyl)-2-methoxybenzamide (6)

[0219] ##STR00033##

[0220] (1R,2R)-2-aminocyclohexane-1-carbonitrile (6-1) (0.1 g, 0.80 mmol) was dissolved in dried toluene (4 mL), and ethyl ((4-(2-(5-chloro-2-methoxybenzamido)ethyl)phenyl)sulfonyl)carbamate (4-1) (0.44 g, 0.96 mmol) was added. The mixture was refluxed at 120° C. for 4 h. The reaction solution was cooled to room temperature and concentrated under reduced pressure to remove the solvent. The residue was purified by pre-HPLC to give compound 6 (43 mg, 96.6% HPLC purity, 10% yield).

[0221] .sup.1HNMR (d6-DMSO, 400 MHz) δ 1.03-1.24 (m, 3H), 1.45-1.58 (m, 4H), 1.95-1.99 (m, 1H), 2.80 (t, J=6.4 Hz, 1H), 2.92 (t, J=6.4 Hz, 2H), 3.52-3.56 (m, 3H), 3.79 (s, 3H), 6.79 (d, J=7.6 Hz, 1H), 7.15 (d, J=8.8 Hz, 1H), 7.44-7.51 (m, 3H), 7.64 (s, 1H), 7.84 (d, J=7.2 Hz, 2H), 8.26 (bs, 1H), 10.93 (bs, 1H).

Example 7: Preparation of 5-cyano-N-(4-(N-(cyclohexylcarbamoyl)sulfamoyl)phenethyl)-2-methoxybenzamide (7)

[0222] ##STR00034##

Step 1: preparation of 5-cyano-2-methoxy-N-(4-sulfamoylphenethyl)benzamide (7-2)

[0223] To a 25 mL reaction flask were added 5-cyano-2-methoxybenzoic acid (7-1) (500 mg, 2.82 mmol), 4-(2-aminoethyl)benzenesulfonamide (2-4) (565.8 mg, 2.82 mmol) and MeCN (2.8 mL) under nitrogen atmosphere. The mixture was stirred and cooled under an ice bath. Pyridine (697 mg, 8.81 mmol) and EDCI (541 mg, 2.82 mmol) were added, and the mixture was stirred for 30 min while maintaining the ice bath. The ice bath was removed, and the mixture was naturally warmed to room temperature and stirred overnight. The reaction solution was concentrated under reduced pressure, and water (2.5 mL) was added. The resulting mixture was stirred for 30 min. The reaction solution was filtered. The solid was collected and dried to give compound 7-2 (934 mg, 92% reaction yield) as an off-white solid, which was directly used in the next step without purification.

Step 2: preparation of 5-cyano-N-(4-(N-(cyclohexylcarbamoyl)sulfamoyl)phenethyl)-2-methoxybenzamide (7)

[0224] To a 25 mL flask were added compound 7-2 (567 mg, 1.578 mmol), isocyanatocyclohexane (2-6) (316 mg, 2.525 mmol), and DMF (5 mL) at room temperature under nitrogen atmosphere. The mixture was dissolved with stirring and cooled under an ice bath, and a solution of KOtBu in THF (2.4 mL, 2.4 mmol) was added dropwise. The resulting mixture was stirred for 10 minutes while maintaining the ice bath. The ice bath was removed, and the mixture was naturally warmed to room temperature and stirred overnight. The reaction system was diluted with water (15 mL), stirred for 5 min and filtered, and the residue was diluted with ethyl acetate, stirred for 30 min and filtered. The solid was washed once with a small amount of ethyl acetate, collected, slurried with DCM:MeOH=10:1 (4 mL) at room temperature for 30 min, and filtered. The solid was collected to give compound 7 (220 mg, 95.9% purity, 28.8% yield).

[0225] .sup.1HNMR (d.sup.6-DMSO, 400 MHz) δ 1.05-1.23 (m, 5H), 1.46-1.65 (m, 5H), 2.93 (t, J=6.4 Hz, 2H), 3.28 (bs, 1H), 3.52-3.56 (m, 2H), 3.87 (s, 3H), 6.34 (d, J=7.6 Hz, 1H), 7.30 (d, J=8.0 Hz, 1H), 7.49 (d, J=7.6 Hz, 2H), 7.84 (d, J=8.0 Hz, 2H), 7.92-7.97 (m, 2H), 8.32 (bs, 1H), 10.32 (bs, 1H).

Biological Assay

Test Example 1: Binding Activity of the Compounds of the Present Disclosure for SUR1 Receptor

[0226]

TABLE-US-00001 Reagent Supplier Cat. No. SUR1 RECEPTOR MEMBRANE JIANGSU HENGRUI MEDICINE CO., LTD. Glibenclamide, Glyburide, [Cyclohexyl- Perkin Elmer NET1024250UC 2,3-3H(N)]-(9.25 MBq) Glipizide MCE HY-B0254 ULTIMA GOLD Perkin Elmer 77-16061 96-well round-bottom deep-well plate, Perkin Elmer P-DW-11-C 1.1 mL 384-well round-bottom plate corning 3657 UNIFILTER-96 GF/B filter plate Perkin Elmer 6005177 Polyethyleneimine (PEI) sigma 408727 Centrifuge tube (15 mL, 50 mL) BD 352096, 352070 Loading slot Jet biofil LTT001050 TIP (10 μL, 200 μL, 1 ml) AXYGEN T-300-R-S T-200-Y-R-S T-1000-B-R-S MgC1.sub.2 sigma 7786-30-3 HEPES sigma 7365-45-9 5′-adenosine triphosphate disodium salt sigma A2383 hydrate/ATP

TABLE-US-00002 Instruments Supplier Model Vortex mixer IKA MS3 BASIC Electric thermostatic Shanghai Yiheng DHP-9032 incubator Microplate shaker VWR 12620-928 TopCount Perkin Elmer NTX Universal Harvester Perkin Elmer UNIFILTER-96

TABLE-US-00003 Reaction buffer Stock solution Final Name concentration Mass/volume concentration HEPES 1M 15 mL 30 mM  MgCl.sub.2 1M 1 mL 2 mM ATP-Na.sub.2 Powder 0.55 g 2 mM H.sub.2O — Making up with water — to 500 mL (500 mL) pH 7.4

TABLE-US-00004 Washing solution Stock solution Final Name concentration Mass/volume concentration NaCl Powder 13.5 g 0.9% 1.5 L

[0227] Procedures: [0228] a) 100 μL of reaction buffer was added to each well of a 96-well deep-well plate. [0229] b) 5 μL of diluted test compound (1% DMSO) was added to each well of the 96-well deep-well plate. [0230] c) 30 μL of SUR1 membrane protein and 270 μL of reaction mixture were added to each well, and the plate was shaken at 600 rpm for 5 min. [0231] d) A mixed solution of 100 μL of reaction buffer and [3H]-Glibenclamide (final concentration of 2 nM) was added to the reaction system, and the plate was shaken at 600 rpm for 5 min and incubated at 37° C. for 1 h. [0232] e) A UNIFILTER-96 GF/B plate was pretreated with 0.5% PEI, and 150 μL of 0.5% PEI was added to each well. The plate was incubated at 4° C. for 1 h. [0233] f) The UNIFILTER-96 GF/C and UNIFILTER-96 GF/B plates were washed twice with Universal Harvester, each time with 50 mL of washing solution. [0234] g) The SUR1 receptor reaction system was transferred to a UNIFILTER-96 GF/B plate with Universal Harvester, and 900 μL of washing solution was added to each well to wash the GF/B plate 4 times. The washed UNIFILTER-96 GF/B plate was dried in an oven at 55° C. for 10 min. [0235] h) 40 μL of ULTIMA GOLD scintillation solution was added to each well, and the plate was read using Microbeta.

[0236] Results: the binding activity of the compounds of the present disclosure for the SUR1 receptor is shown in Table 1 below.

TABLE-US-00005 TABLE 1 Binding activity of the compounds of the present disclosure for the SUR1 receptor Compound IC.sub.50 (nM) Glipizide 229.592 Glibenclamide 6 Compound 1 12.695 Compound 2 9 Compound 3 159 Compound 4 8 Compound 5 9 Compound 6 37 Compound 7 6

Test Example 2: Assay on In Vivo Pharmacokinetics and Brain Tissue Distribution of Different Compounds Administered by Intravenous Injection in SD Rats

Preparation of Test Sample

[0237] 1. An appropriate amount of compound 1, compound 4, compound 7 or glibenclamide was accurately weighed, and an appropriate volume of 5% DMSO+10% Solutol+85% normal saline was added thereto. The mixture was stirred or sonicated until completely dissolved to obtain a clear solution at a concentration of 1 mg/mL for administration by intravenous injection.

[0238] 2. An appropriate amount of compound 3 or compound 6 was accurately weighed, and an appropriate volume of 5% DMSO+20% PG+20% PEG400+55% PBS (pH 8.0) was added thereto. The mixture was stirred or sonicated until completely dissolved to obtain a clear solution at a concentration of 1 mg/mL for administration by intravenous administration.

[0239] Experimental Animals

[0240] SPF grade SD rats, source: animals were transferred from the laboratory animal reserve bank (999M-017), Shanghai Sippe-Bk Lab Animal Co., Ltd.

TABLE-US-00006 Design of experiment Volume for Test Number Dose Concentration administration Route of Group compounds Male** (mg/kg) (mg/mL) (mL/kg) administration Sample type 1 Compound 1 6 5 1 5 IV Blood and tissue samples 2 Compound 3 6 5 1 5 IV Blood and tissue samples 3 Compound 4 6 5 1 5 IV Blood and tissue samples 4 Compound 6 6 5 1 5 IV Blood and tissue samples 5 Compound 7 6 5 1 5 IV Blood and tissue samples 6 Glibenclamide 6 5 1 5 IV Blood and tissue samples

[0241] Collection Time Point

[0242] 3 rats/time point was set, blood samples were collected from the first 3 rats in each group, and cerebrospinal fluid and brain tissue were collected from the last 3 rats in each group. Blood sample: before administration and 5 min, 0.25 h, 0.5 h, 1 h, 1.5 h, 2 h, 4 h, 6 h, 8 h, 12 h and 24 h after administration. Tissue sample: cerebrospinal fluid and brain tissue were collected 10 min after administration.

[0243] Sample Collection and Processing

[0244] Blood sample: blood was collected via jugular vein or other suitable means, and about 0.20 mL of blood was collected from each rat and added into an EDTA-K2 anticoagulation tube, which was then placed on ice after collection and centrifuged within 2 h for plasma separation (centrifugation conditions: 6800 g, 6 min, 2-8° C.). The collected plasma samples were stored in a refrigerator at −70° C. before analysis, and the residual plasma samples after analysis were still stored in the refrigerator at −70° C. for subsequent processing according to the requirements of the sponsor.

[0245] Tissue sample: cerebrospinal fluid and brain tissues were collected from animals used for tissue collection 10 min after administration, and the brain tissues were separately washed with normal saline to avoid cross contamination, dried with filter paper, and weighed. The tissues were then placed in marked tubes (one tube for each tissue), and the samples were temporarily placed on ice prior to storage in a refrigerator at −70° C.

[0246] The collected plasma samples and the collected tissue samples were stored in a refrigerator at −70° C. before analysis, and the residual plasma samples and the residual tissue samples after analysis were still stored in the refrigerator at −70° C. for subsequent processing according to the requirements of the sponsor.

[0247] Biological Analysis and Data Processing

[0248] The concentration of each test compound in the plasma and tissue samples was determined. The accuracy of quality control samples was evaluated while analyzing the samples, and more than 66% of the quality control samples were required to have an accuracy between 80% and 120%.

[0249] The pharmacokinetic parameters such as AUC(0-t), T½, Cmax, Tmax and MRT were calculated from plasma concentration data at different time points using WinNonlin.

[0250] When plasma drug concentration-time curves were plotted, BLQ was recorded as 0. When the pharmacokinetic parameters were calculated, the concentration before administration was calculated as 0; BLQ before Cmax (including “No peak”) was calculated as 0; and BLQ that occurs after Cmax (including “No peak”) was excluded from the calculation.

TABLE-US-00007 TABLE 2 Changes in the concentration of the compounds in plasma after a single intravenous injection Compound 1 Compound 3 Compound 4 Compound 6 Compound 7 Glibenclamide T.sub.1/2 (h)  3.9 ± 0.6  2.6 ± 0.1 3.1 ± 3.4 5.4 ± 2.1  1.9 ± 0.2  2.5 ± 0.1 T.sub.max (h) 0.1 ± 0  0.1 ± 0  0.1 ± 0.sup.  0.1 ± 0.sup.  0.1 ± 0   0.1 ± 0 C.sub.max 64998.1 ± 3633.1 32785.3 ± 1528.9 39579.7 ± 4734.8  11788.2 ± 1063.3  42446.6 ± 8443.2 36629.6 ± 682.2 (ng/ml) AUC.sub.(0-t) .sup. 28194 ± 5043.5 35659.2 ± 1128.5 12941.9 ± 1457.5  4655.9 ± 259.3  28485.9 ± 7925.3 23608.2 ± 688.1 (h* ng/ml) AUC.sub.(0-∞) 28737.1 ± 4700.2 36400.5 ± 1168.2  12980 ± 1478.6 4981.9 ± 288   28561.4 ± 7915.1 23829.9 ± 705.3 (h* ng/ml) MRT.sub.(0-t)(h)  0.9 ± 0.3  1.9 ± 0.1 0.3 ± 0.1 0.7 ± 0.1  1 ± 0  1.1 ± 0.1 MRT.sub.(0-∞)  1.2 ± 0.1  2.2 ± 0.1 0.3 ± 0.1 3.6 ± 4.8 1.1 ± 0   1.2 ± 0.1 (h) Vss (ml/kg) 219.1 ± 26.3 303.5 ± 14.1 118.6 ± 12.3  3439.6 ± 4529.2 192.7 ± 46.9 253.4 ± 20  Vz (ml/kg) 1000.9 ± 202.4 509.8 ± 30.7 1655.1 ± 1733.9 14571.4 ± 19059.9  499.9 ± 160.3  768.9 ± 16.8 Note: the dose is 5 mg/kg for all the test compounds; and the lower limit of quantitation varies from 5 to 30 ng/ml.