6-OXO-1,6-DIHYDROPYRIDAZINE DERIVATIVE, PREPARATION METHOD THEREFOR AND MEDICAL USE THEREOF
20220073471 · 2022-03-10
Inventors
- Fanglong Yang (Shanghai, CN)
- Nan Yu (Shanghai, CN)
- Jiangtao CHI (Shanghai, CN)
- Feng He (Shanghai, CN)
- Weikang Tao (Shanghai, CN)
Cpc classification
A61P29/00
HUMAN NECESSITIES
International classification
Abstract
A 6-oxo-1,6-dihydropyridazine derivative, a preparation method therefor and medical use thereof, in particular, a 6-oxo-1,6-dihydropyridazine derivative represented by general formula (I), a preparation method therefor, and a pharmaceutical composition containing the derivative, and use thereof as a Nav inhibitor and use thereof in the preparation of a drug for the treatment and/or prevention of pain and pain-related diseases. Each substituent in general formula (I) is the same as defined in the description.
##STR00001##
Claims
1. A compound of formula (I) or a tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof: ##STR00149## wherein: M is selected from the group consisting of O atom, CR.sup.4R.sup.5 and S atom; ring A is an aryl or heteroaryl, wherein the aryl or heteroaryl is optionally fused to a cycloalkyl or heterocyclyl; each R.sup.1 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, deuterated alkyl, deuterated alkoxy, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; each R.sup.2 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, deuterated alkyl, deuterated alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, cycloalkyloxy, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each optionally substituted by one or more substituents selected from the group consisting of alkyl, haloalkyl, halogen, amino, nitro, cyano, hydroxy, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; each R.sup.3 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.4 and R.sup.5 are identical or different and are each independently selected from the group consisting of hydrogen atom, deuterium atom, halogen, alkyl, alkoxy, haloalkyl, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; n is 0, 1, 2, 3 or 4; s is 0, 1, 2, 3 or 4; and t is 0, 1 or 2.
2. The compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein ring A is selected from the group consisting of phenyl, ##STR00150## and pyridyl.
3. The compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein M is selected from the group consisting of O atom, CH.sub.2 and S atom.
4. The compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, being a compound of formula (II) or a tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof: ##STR00151## wherein: R.sup.1, R.sup.2, R.sup.3, n, s and t are as defined in claim 1.
5. The compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, being a compound of formula (III) or a tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof: ##STR00152## wherein: M is selected from the group consisting of O atom, CH.sub.2 and S atom; R.sup.1a is a halogen; R.sup.1b is selected from the group consisting of halogen, alkyl, alkoxy, haloalkyl and haloalkoxy; each R.sup.2 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, deuterated alkyl, deuterated alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, cycloalkyloxy, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each optionally substituted by one or more substituents selected from the group consisting of alkyl, haloalkyl, halogen, amino, nitro, cyano, hydroxy, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; each R.sup.3 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; s is 0, 1, 2, 3 or 4; and t is 0, 1 or 2.
6. The compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein each R.sup.1 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl and haloalkoxy.
7. The compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein each R.sup.2 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, deuterated alkyl, alkoxy, deuterated alkoxy, hydroxy, haloalkyl, haloalkoxy, cycloalkyl and cycloalkyloxy.
8. The compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein s is 2.
9. The compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, being a compound of formula (IV) or a tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof: ##STR00153## wherein: R.sup.1a is a halogen; R.sup.1b is selected from the group consisting of halogen, alkyl, alkoxy, haloalkyl and haloalkoxy; R.sup.2a is an alkoxy or deuterated alkoxy; R.sup.2b is selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy and haloalkoxy; each R.sup.3 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; and t is 0, 1 or 2.
10. The compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.3 is a hydrogen atom.
11. The compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, selected from the group consisting of: ##STR00154## ##STR00155## ##STR00156## ##STR00157## ##STR00158##
12. A compound of formula (IA) or a tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof: ##STR00159## wherein: X is a halogen; M is selected from the group consisting of O atom, CR.sup.4R.sup.5 and S atom; ring A is an aryl or heteroaryl, wherein the aryl or heteroaryl is optionally fused to a cycloalkyl or heterocyclyl; each R.sup.1 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, deuterated alkyl, deuterated alkoxy, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; each R.sup.2 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, deuterated alkyl, deuterated alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, cycloalkyloxy, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each optionally substituted by one or more substituents selected from the group consisting of alkyl, haloalkyl, halogen, amino, nitro, cyano, hydroxy, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; each R.sup.3 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.4 and R.sup.5 are identical or different and are each independently selected from the group consisting of hydrogen atom, deuterium atom, halogen, alkyl, alkoxy, haloalkyl, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; n is 0, 1, 2, 3 or 4; s is 0, 1, 2, 3 or 4; and t is 0, 1 or 2.
13. The compound of formula (IA) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 12, selected from the group consisting of: ##STR00160## ##STR00161##
14. A compound of formula (IB) or a tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof: ##STR00162## wherein: Y is a halogen; each R.sup.1 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, deuterated alkyl, deuterated alkoxy, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; each R.sup.3 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; n is 0, 1, 2, 3 or 4; and t is 0, 1 or 2.
15. The compound of formula (IB) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 14, selected from the group consisting of: ##STR00163##
16. A method for preparing the compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, comprising a step of: ##STR00164## reacting a compound of formula (IA) to obtain the compound of formula (I); wherein: X is a halogen; and M is selected from the group consisting of O atom, CR.sup.4R.sup.5 and S atom; ring A is an aryl or heteroaryl, wherein the aryl or heteroaryl is optionally fused to a cycloalkyl or heterocyclyl; each R.sup.1 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, deuterated alkyl, deuterated alkoxy, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; each R.sup.2 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, deuterated alkyl, deuterated alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, cycloalkyloxy, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each optionally substituted by one or more substituents selected from the group consisting of alkyl, haloalkyl, halogen, amino, nitro, cyano, hydroxy, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; each R.sup.3 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.4 and R.sup.5 are identical or different and are each independently selected from the group consisting of hydrogen atom, deuterium atom, halogen, alkyl, alkoxy, haloalkyl, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; n is 0, 1, 2, 3 or 4; s is 0, 1, 2, 3 or 4; and t is 0, 1 or 2.
17. A method for preparing the compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, comprising a step of: ##STR00165## reacting a compound of formula (IB) and a compound of formula (IC) to obtain the compound of formula (I); wherein: Y is a halogen; M is selected from the group consisting of O atom, CR.sup.4R.sup.5 and S atom; ring A is an aryl or heteroaryl, wherein the aryl or heteroaryl is optionally fused to a cycloalkyl or heterocyclyl; each R.sup.1 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, deuterated alkyl, deuterated alkoxy, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; each R.sup.2 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, deuterated alkyl, deuterated alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, cycloalkyloxy, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each optionally substituted by one or more substituents selected from the group consisting of alkyl, haloalkyl, halogen, amino, nitro, cyano, hydroxy, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; each R.sup.3 is identical or different and each is independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.4 and R.sup.5 are identical or different and are each independently selected from the group consisting of hydrogen atom, deuterium atom, halogen, alkyl, alkoxy, haloalkyl, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; n is 0, 1, 2, 3 or 4; s is 0, 1, 2, 3 or 4; and t is 0, 1 or 2.
18. A pharmaceutical composition, comprising the compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, and one or more pharmaceutically acceptable carriers, diluents or excipients.
19. A method of inhibiting a voltage-gated sodium channel in a subject in need thereof, the method comprising: administering the compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof according to claim 1 to the subject.
20. A method of treating and/or alleviating pain and pain-related diseases, multiple sclerosis, Charcot-Marie-Tooth syndrome, incontinence or cardiac arrhythmia in a subject in need thereof, the method comprising: administering the compound of formula (I) or the tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or mixture thereof of claim 1 to the subject.
Description
DETAILED DESCRIPTION
[0258] The present disclosure will be further described with reference to the following examples, but the examples should not be considered as limiting the scope of the present disclosure.
EXAMPLES
[0259] The structures of the compounds were identified by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS). NMR shifts (6) are given in 10.sup.−6 (ppm). NMR is determined by a Bruker AVANCE-400 machine. The solvents for determination are deuterated-dimethyl sulfoxide (DMSO-d.sub.6), deuterated-chloroform (CDCl.sub.3) and deuterated-methanol (CD.sub.3OD), and the internal standard is tetramethylsilane (TMS).
[0260] MS was determined by a FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Thermo, type: Finnigan LCQ advantage MAX).
[0261] High performance liquid chromatography (HPLC) was determined on an Agilent HPLC 1200DAD, Agilent HPLC 1200VWD and Waters HPLC e2695-2489 high pressure liquid chromatograph.
[0262] Chiral HPLC was determined on an Agilent 1260 DAD high performance liquid chromatograph.
[0263] Preparative chromatography was carried out on Waters 2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP and Gilson-281 preparative chromatographs.
[0264] Chiral preparation was carried out on a Shimadzu LC-20AP preparative chromatograph.
[0265] CombiFlash rapid preparation instrument used was Combiflash Rf200 (TELEDYNE ISCO).
[0266] Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate was used as the thin-layer silica gel chromatography (TLC) plate. The dimension of the silica gel plate used in TLC was 0.15 mm to 0.2 mm, and the dimension of the silica gel plate used in product purification was 0.4 mm to 0.5 mm.
[0267] Yantai Huanghai 200 to 300 mesh silica gel was generally used as a carrier for silica gel column chromatography.
[0268] The average kinase inhibition rates and IC.sub.50 values were determined by a NovoStar microplate reader (BMG Co., Germany).
[0269] The known starting materials of the present disclosure can be prepared by the known methods in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organnics, Aldrich Chemical Company, Accela ChemBio Inc., Dari Chemical Company etc.
[0270] Unless otherwise stated, the reactions were carried out under argon atmosphere or nitrogen atmosphere.
[0271] “Argon atmosphere” or “nitrogen atmosphere” means that a reaction flask is equipped with an argon or nitrogen balloon (about 1 L).
[0272] “Hydrogen atmosphere” means that a reaction flask is equipped with a hydrogen balloon (about 1 L).
[0273] Pressurized hydrogenation reaction was performed on a Parr 3916EKX hydrogenation instrument and a Qinglan QL-500 hydrogen generator or HC2-SS hydrogenation instrument.
[0274] In hydrogenation reactions, the reaction system was generally vacuumed and filled with hydrogen, and the above operation was repeated three times.
[0275] CEM Discover-S 908860 type microwave reactor was used in microwave reactions.
[0276] Unless otherwise stated, the solution refers to an aqueous solution.
[0277] Unless otherwise stated, the reaction temperature is room temperature from 20° C. to 30° C.
[0278] The reaction process in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reactions, the eluent system in column chromatography and the developing solvent system in thin layer chromatography for purification of the compounds included: A: dichloromethane/methanol system, B: n-hexane/ethyl acetate system, C: petroleum ether/ethyl acetate system, D: acetone, E: dichloromethane/acetone system, F: ethyl acetate/dichloromethane system, G: ethyl acetate/dichloromethane/n-hexane, and H: ethyl acetate/dichloromethane/acetone. The ratio of the volume of the solvent was adjusted according to the polarity of the compounds, and a small quantity of alkaline reagent such as triethylamine or acidic reagent such as acetic acid could also be added for adjustment.
Example 1
4,5-Dichloro-2-(4-fluoro-2-methoxyphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide 1
[0279] ##STR00096## ##STR00097##
Step 1
4,5-Dichloro-2-fluorobenzoyl chloride 1b
[0280] Compound 4,5-dichloro-2-fluorobenzoic acid 1a (1.5 g, 7.18 mmol, Accela ChemBio (Shanghai) Inc.) was dissolved in thionyl chloride (10 mL), and the reaction solution was reacted at 80° C. for 16 hours. The reaction solution was concentrated under reduced pressure to obtain the title compound 1b (1.6 g), which was used directly in the next step without purification.
Step 2
4,5-Dichloro-N-(6-chloropyridazin-4-yl)-2-fluorobenzamide 1c
[0281] The crude compound 1b (1.6 g, 7.03 mmol) and 6-chloropyridazine-4-amine (500 mg, 3.86 mmol, Pharmablock Sciences (Nanjing), Inc.) were dissolved in pyridine (10 mL), and the reaction solution was stirred for 16 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 1c (650 mg, yield: 53%) as a white solid.
[0282] MS m/z (ESI): 321.9 [M+1]
Step 3
4,5-Dichloro-N-(6-chloropyridazin-4-yl)-2-(4-fluoro-2-methoxyphenoxy)benzamide 1d
[0283] Compound 1c (100 mg, 0.31 mmol), 4-fluoro-2-methoxyphenol (50 mg, 0.35 mmol, Accela ChemBio (Shanghai) Inc.) and cesium carbonate (153 mg, 0.47 mmol) were added to N,N-dimethylformamide (10 mL), and the reaction solution was reacted at 100° C. for 2 hours. The reaction solution was cooled and filtered through diatomaceous earth. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 1d (100 mg, yield: 72%).
[0284] MS m/z (ESI): 443.7 [M+1]
Step 4
4,5-Dichloro-2-(4-fluoro-2-methoxyphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide 1
[0285] Compound 1d (100 mg, 0.22 mmol) and potassium acetate (45 mg, 0.46 mmol) were added to acetic acid (5 mL), and the reaction solution was reacted at 120° C. for 1.5 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 1 (70 mg, yield: 73%).
[0286] MS m/z (ESI): 425.8 [M+1]
[0287] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.81 (s, 1H), 10.85 (s, 1H), 7.93 (s, 1H), 7.91 (d, 1H), 7.25-7.20 (m, 1H), 7.11 (dd, 1H), 6.88 (s, 1H), 6.82-6.82 (m, 1H), 5.73 (s, 1H), 3.73 (s, 3H).
Example 2
5-Chloro-2-(4-fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 2
[0288] ##STR00098##
Step 1
5-Chloro-2-fluoro-4-(trifluoromethyl)benzoic acid 2b
[0289] 2,2,6,6-Tetramethylpiperidine (19.2 g, 135.93 mmol, Accela ChemBio (Shanghai) Inc.) was added to tetrahydrofuran (200 mL) under an argon atmosphere. The reaction solution was cooled to 0° C., then n-butyl lithium (1.6 M, 85.1 mL) was added dropwise within about 45 minutes at a controlled temperature below 3° C. The reaction solution was reacted at 0° C. for 1 hour, and then cooled to −78° C. Compound 1-chloro-4-fluoro-2-(trifluoromethyl)benzene 2a (18 g, 90.66 mmol, Shanghai Titan Scientific Co., Ltd.) was added dropwise, and the reaction solution was reacted for 3 hours. Excess dry ice was added, and the reaction solution was naturally warmed up to 0° C., followed by the addition of 150 mL of ice water. The reaction solution was separated into two phases. The aqueous phase was adjusted to pH 5 to 6 with concentrated hydrochloric acid and extracted with ethyl acetate (50 mL), and the organic phase was concentrated under reduced pressure. The crude product was washed with n-hexane (50 mL), then purified by silica gel column chromatography with eluent system A to obtain the title compound 2b (15 g, yield: 68%).
[0290] MS m/z (ESI): 241.1 [M−1]
Step 2
Methyl 5-chloro-2-fluoro-4-(trifluoromethyl)benzoate 2c
[0291] Compound 2b (5 g, 20.61 mmol) was added to thionyl chloride (49.2 g, 413.55 mmol), and the reaction solution was reacted at 80° C. for 2 hours. The reaction solution was concentrated under reduced pressure. The resulting oil was added dropwise to methanol (100 mL), and the reaction solution was reacted at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 2c (2.78 g, yield: 52%).
Step 3
Methyl 5-chloro-2-(4-fluoro-2-methylphenoxy)-4-(trifluoromethyl)benzoate 2d
[0292] Compound 2c (2.78 g, 10.83 mmol), 4-fluoro-2-methyl-phenol (1.5 g, 11.89 mmol, Shanghai Bide Pharmatech Ltd.) and cesium carbonate (6 g, 18.41 mmol) were added to N,N-dimethylformamide (20 mL), and the reaction solution was reacted at 100° C. for 1 hour. The reaction solution was cooled and filtered. The filtrate was concentrated to obtain the target compound 2d (3.92 g), which was used directly in the next step without purification.
[0293] MS m/z (ESI): 363.1 [M+1]
Step 4
5-Chloro-2-(4-fluoro-2-methylphenoxy)-4-(trifluoromethyl)benzoic acid 2e
[0294] Compound 2d (3.92 g, 10.81 mmol) was dissolved in methanol (30 mL), followed by the addition of water (10 mL) and sodium hydroxide (1.3 g, 32.5 mmol), and the reaction solution was reacted for 16 hours. The reaction solution was concentrated, followed by the addition of 10 mL of water, and the pH was adjusted to 1 with concentrated hydrochloric acid. The resulting solution was extracted with ethyl acetate (20 mL×3), and the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 2e (3.67 g, yield: 97%).
[0295] MS m/z (ESI): 346.8 [M−1]
Step 5
5-Chloro-2-(4-fluoro-2-methylphenoxy)-4-(trifluoromethyl)benzoyl chloride 2f
[0296] Compound 2e (3.67 g, 10.52 mmol) was added to thionyl chloride (20 g, 168.1 mmol), and the reaction solution was reacted at 80° C. for 2 hours. The reaction solution was concentrated to obtain the title compound 2f (3.86 g), which was used directly in the next step without purification.
Step 6
5-Chloro-N-(6-chloropyridazin-4-yl)-2-(4-fluoro-2-methylphenoxy)-4-(trifluoromethyl)benzamide 2g
[0297] 4-Dimethylaminopyridine (130 mg, 1.05 mmol) and 6-chloropyridazin-4-amine (1.51 g, 11.57 mmol, Pharmablock Sciences (Nanjing), Inc.) were dissolved in pyridine (40 mL), and the resulting solution was dried over molecular sieves. Compound 2f (3.86 g, 10.51 mmol) was added, and the reaction solution was reacted for 16 hours under an argon atmosphere. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 2g (1.3 g, yield: 39%).
[0298] MS m/z (ESI): 460.0 [M+1]
Step 7
5-Chloro-2-(4-fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 2
[0299] Compound 2g (1.3 g, 2.82 mmol) and potassium acetate (555 mg, 5.65 mmol) were added to acetic acid (20 mL), and the reaction solution was reacted at 130° C. for 3 hours. The reaction solution was concentrated, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 2 (800 mg, yield: 64%).
[0300] MS m/z (ESI): 442.0 [M+1]
[0301] 1H NMR (400 MHz, DMSO-d.sub.6) δ 12.83 (s, 1H), 11.03 (s, 1H), 8.07 (s, 1H), 7.86 (s, 1H), 7.05-7.25 (m, 5H), 2.14 (s, 3H).
Example 3
4,5-Dichloro-2-(4-fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide 3
[0302] ##STR00099##
[0303] In accordance with the synthetic route in Example 1, the starting compound 4-fluoro-2-methoxyphenol in Step 3 was replaced with compound 4-fluoro-2-methylphenol, accordingly, the title compound 3 (20 mg) was prepared.
[0304] MS m/z (ESI): 407.8 [M+1]
[0305] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.82 (s, 1H), 10.92 (s, 1H), 7.99 (s, 1H), 7.90 (d, 1H), 7.21-7.18 (m, 2H), 7.08-7.05 (m, 2H), 6.99 (s, 1H), 2.14 (s, 3H).
Example 4
4,5-Dichloro-2-(4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide 4
[0306] ##STR00100##
[0307] In accordance with the synthetic route in Example 1, the starting compound 4-fluoro-2-methoxyphenol in Step 3 was replaced with compound 4-fluorophenol, accordingly, the title compound 4 (55 mg) was prepared.
[0308] MS m/z (ESI): 395.8 [M+1]
[0309] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.81 (s, 1H), 10.89 (s, 1H), 7.98 (s, 1H), 7.88 (d, 1H), 7.26-7.22 (m, 2H), 7.19 (s, 1H), 7.16-7.13 (m, 3H).
Example 5
2-(4-Fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(perfluoroethyl)benzamide 5
[0310] ##STR00101## ##STR00102##
Step 1
2-Fluoro-4-(perfluoroethyl)benzoic acid 5b
[0311] 4-Bromo-2-fluorobenzoic acid 5a (2.19 g, 10 mmol, J&K Scientific Ltd.) was dissolved in dimethyl sulfoxide (41 mL) under an argon atmosphere. Copper powder (6.36 g, 100 mmol) and pentafluoroiodoethane (17.22 g, 70 mmol, 8.24 mL, Sichuan Shang Fluoro Technology Co., Ltd.) were added, and the reaction solution was sealed in a tube and reacted at 120° C. for 72 hours. The reaction solution was cooled, followed by the addition of 100 mL of water and 100 mL of ethyl acetate, stirred well and filtered. The filtrate was separated into two phases, and the aqueous phase was extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 5b (2.4 g, yield: 93%).
[0312] MS m/z (ESI): 257.0 [M−1]
Step 2
2-Fluoro-4-(perfluoroethyl)benzoyl chloride 5c
[0313] Compound 5b (500 mg, 1.93 mmol) was added to thionyl chloride (8.19 g, 68.8 mmol), and the reaction solution was reacted at 80° C. for 16 hours. The reaction solution was concentrated to obtain the crude title compound 5c (535 mg), which was used directly in the next step without purification.
Step 3
N-(6-Chloropyridazin-4-yl)-2-fluoro-4-(perfluoroethyl)benzamide 5d
[0314] The crude compound 5c (535 mg, 1.93 mmol), 6-chloropyridazine-4-amine (150 mg, 1.16 mmol) and pyridine (916 mg, 11.58 mmol) were dissolved in dichloromethane (5 mL), and the reaction solution was reacted for 16 hours. 50 mL ethyl acetate was added, and the reaction solution was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 5d (664 mg, yield: 93%).
[0315] MS m/z (ESI): 370.0 [M+1]
Step 4
2-Fluoro-N-(6-oxo-1,6-dihydropyridazin)-4-(perfluoroethyl)benzamide 5e
[0316] Compound 5d (400 mg, 1.08 mmol) and potassium acetate (213 mg, 2.17 mmol) were added to acetic acid (5 mL), and the reaction solution was reacted at 130° C. for 5 hours. The reaction solution was concentrated, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 5e (190 mg, yield: 50%).
[0317] MS m/z (ESI): 352.0 [M+1]
Step 5
2-(4-Fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(perfluoroethyl)benzamide 5
[0318] Compound 5e (190 mg, 0.54 mmol), 4-fluoro-2-methylphenol (103 mg, 0.82 mmol) and cesium carbonate (265 mg, 0.81 mmol) were added to N,N-dimethylformamide (3 mL), and the reaction solution was reacted at 100° C. for 2 hours. The reaction solution was cooled, filtered and purified by preparative high performance liquid chromatography (Waters 2767-SQ Detecor2, eluent system: ammonium bicarbonate, water, acetonitrile) to obtain the title compound 5 (89 mg, yield: 36%).
[0319] MS m/z (ESI): 458.1 [M+1]
[0320] .sup.1H NMR (400 MHz, CD.sub.3OD): δ 8.05 (d, 1H), 7.92 (d, 1H), 7.52 (d, 1H), 7.49 (d, 1H), 7.12 (dd, 1H), 6.99-7.07 (m, 2H), 6.93 (s, 1H), 2.21 (s, 3H).
Example 6
2-(2-Methyl-4-(trifluoromethoxy)phenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 6
[0321] ##STR00103## ##STR00104##
Step 1
2-Fluoro-4-(trifluoromethyl)benzoyl chloride 6b
[0322] 2-Fluoro-4-(trifluoromethyl)benzoic acid 6a (1.2 g, 5.76 mmol, Accela ChemBio (Shanghai) Inc.) was added to thionyl chloride (12 mL), and the reaction solution was reacted at 80° C. for 16 hours. The reaction solution was concentrated to obtain the crude title compound 6b (1.3 g), which was used directly in the next step without purification.
Step 2
N-(6-Chloropyridazin-4-yl)-2-fluoro-4-(trifluoromethyl)benzamide 6c
[0323] The crude compound 6b (1.3 g, 5.74 mmol), 6-chloro-4-aminopyridazine (300 mg, 2.31 mmol) and pyridine (916 mg, 11.58 mmol) were dissolved in dichloromethane (5 mL), and the reaction solution was reacted for 16 hours. The reaction solution was concentrated, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 6c (475 mg, yield: 64%).
[0324] MS m/z (ESI): 320.0 [M+1]
Step 3
2-Fluoro-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 6d
[0325] Compound 6c (475 mg, 1.49 mmol) and potassium acetate (292 mg, 2.98 mmol) were added to acetic acid (5 mL), and the reaction solution was reacted at 120° C. for 16 hours. The reaction solution was concentrated, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 6d (300 mg, yield: 67%).
[0326] MS m/z (ESI): 302.0 [M+1]
Step 4
2-(2-Methyl-4-(trifluoromethoxy)phenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 6
[0327] Compound 6d (100 mg, 0.33 mmol), 2-methyl-4-(trifluoromethoxy)phenol (78 mg, 0.4 mmol) and cesium carbonate (216 mg, 0.66 mmol) were added to N,N-dimethylformamide (3 mL), and the reaction solution was reacted at 100° C. for 8 hours. The reaction solution was cooled and filtered. The filtrate was concentrated, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 6 (60 mg, yield: 38%).
[0328] MS m/z (ESI): 473.8 [M+1]
[0329] .sup.1H NMR (400 MHz, CD.sub.3OD): δ 8.05 (d, 1H), 7.92 (d, 1H), 7.52 (d, 1H), 7.49 (d, 1H), 7.12 (dd, 1H), 6.99-7.07 (m, 2H), 6.93 (s, 1H), 2.29 (s, 3H).
Example 7
2-(4-Fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-5-(trifluoromethyl)benzamide 7
[0330] ##STR00105## ##STR00106##
Step 1
2-Fluoro-5-(trifluoromethyl)benzoyl chloride 7b
[0331] 2-Fluoro-5-(trifluoromethyl)benzoic acid 7a (382 mg, 1.83 mmol, Shanghai Bide Pharmatech Ltd.) was added to thionyl chloride (5 mL), and the reaction solution was reacted at 80° C. for 16 hours. The reaction solution was concentrated to obtain the crude title compound 7b (400 mg), which was used directly in the next step without purification.
Step 2
N-(6-Chloropyridazin-4-yl)-2-fluoro-5-(trifluoromethyl)benzamide 7c
[0332] The crude compound 7b (393 mg, 1.73 mmol), 6-chloro-4-aminopyridazine (150 mg, 1.15 mmol) and pyridine (458 mg, 5.79 mmol) were dissolved in dichloromethane (5 mL), and the reaction solution was reacted for 24 hours. 50 mL ethyl acetate was added, and the reaction solution was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 7c (90 mg, yield: 24%).
[0333] MS m/z (ESI): 320.0 [M+1]
Step 3
N-(6-Chloropyridazin-4-yl)-2-(4-fluoro-2-methylphenoxy)-5-(trifluoromethyl)benzamide 7d
[0334] Compound 7c (90 mg, 0.28 mmol), 4-fluoro-2-methylphenol (43 mg, 0.34 mmol) and cesium carbonate (183 mg, 0.56 mmol) were added to N,N-dimethylformamide (3 mL), and the reaction solution was reacted at 100° C. for 1 hour. The reaction solution was filtered, and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 7d (46 mg, yield: 37%).
[0335] MS m/z (ESI): 426.1 [M+1]
Step 4
2-(4-Fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-5-(trifluoromethyl)benzamide 7
[0336] Compound 7d (46 mg, 0.11 mmol) and potassium acetate (21 mg, 0.21 mmol) were added to acetic acid (1 mL), and the reaction solution was reacted at 130° C. for 4 hours. The reaction solution was concentrated, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 7 (16 mg, yield: 36%).
[0337] MS m/z (ESI): 407.9 [M+1]
[0338] .sup.1H NMR (400 MHz, CD.sub.3OD): δ 8.08-8.07 (m, 2H), 7.77 (dd, 1H), 7.52 (d, 1H), 7.14-7.11 (m, 2H), 7.05-7.00 (m, 1H), 6.89 (d, 1H), 2.21 (s, 3H).
Example 8
2-(4-Fluoro-2-methoxyphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 8
[0339] ##STR00107##
[0340] In accordance with the synthetic route in Example 6, the starting compound 2-methyl-4-(trifluoromethoxy)phenol in Step 3 was replaced with compound 4-fluoro-2-methoxyphenol, accordingly, the title compound 8 (20 mg) was prepared.
[0341] MS m/z (ESI): 424.1 [M+1]
[0342] .sup.1H NMR (400 MHz, CD.sub.3OD): δ 8.07 (d, 1H), 7.94 (d, 1H), 7.53-7.49 (m, 2H), 7.26 (q, 1H), 7.03 (dd, 1H), 6.97 (s, 1H), 6.83-6.79 (m, 1H), 2.89 (s, 3H).
Example 9
2-(4-Fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 9
[0343] ##STR00108##
[0344] In accordance with the synthetic route in Example 6, the starting compound 2-methyl-4-(trifluoromethoxy)phenol in Step 3 was replaced with compound 4-fluoro-2-methylphenol, accordingly, the title compound 9 (65 mg) was prepared.
[0345] MS m/z (ESI): 408.1 [M+1]
[0346] .sup.1H NMR (400 MHz, CD.sub.3OD): δ 8.05 (d, 1H), 7.90 (d, 1H), 7.54 (d, 1H), 7.49 (d, 1H), 7.11 (dd, 1H), 6.99-7.07 (m, 2H), 6.97 (s, 1H), 2.22 (s, 3H).
Example 10
N-(6-Oxo-1,6-dihydropyridazin-4-yl)-2-(4-(trifluoromethoxy)phenoxy)-4-(trifluoromethyl)benzamide 10
[0347] ##STR00109##
[0348] In accordance with the synthetic route in Example 6, the starting compound 2-methyl-4-(trifluoromethoxy)phenol in Step 3 was replaced with compound 4-trifluoromethoxyphenol, accordingly, the title compound 10 (18 mg) was prepared.
[0349] MS m/z (ESI): 460.0 [M+1]
[0350] .sup.1H NMR (400 MHz, CD.sub.3OD): δ 8.03 (d, 1H), 7.93 (d, 1H), 7.65-7.63 (m, 1H), 7.43 (d, 1H), 7.35-7.32 (m, 3H), 7.19-7.16 (m, 2H).
Example 11
5-Chloro-2-(4-fluoro-2-methoxyphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 11
[0351] ##STR00110##
Step 1
5-Chloro-2-fluoro-4-(trifluoromethyl)benzoyl chloride 11a
[0352] Compound 2b (5.00 g, 20.6 mmol) was added to 15 mL of thionyl chloride, and the reaction solution was reacted at 80° C. for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the crude title compound 11a (5.38 g), which was used directly in the next step without purification.
Step 2
5-Chloro-2-fluoro-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 11b
[0353] 5-Aminopyridazin-3-one (3.06 g, 24.8 mmol, Shanghai Medicilon Inc.) was dissolved in 40 mL of N-methylpyrrolidone. The resulting solution was cooled to 0° C., and sodium hydride (2.06 g, 51.5 mmol, purity: 60%) was slowly add in batches. The reaction solution was stirred at 0° C. for 30 minutes. Compound 11a (5.38 g, 20.6 mmol) was dissolved in 3 mL of N-methylpyrrolidone, and the resulting solution was slowly added dropwise to the above reaction solution, which was then stirred at room temperature overnight. The reaction solution containing the title compound 11b was used directly in the next step without purification.
Step 3
5-Chloro-2-(4-fluoro-2-methoxyphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 11c
[0354] 4-Fluoro-2-methoxyphenol (2.34 g, 16.5 mmol, Tokyo Chemical Industry (Shanghai) Co., Ltd.) and cesium carbonate (6.71 g, 20.6 mmol, Accela ChemBio (Shanghai) Inc.) were added directly to the reaction solution containing compound 11b. The reaction solution was reacted at 60° C. overnight, and then cooled to room temperature. Ethyl acetate (250 mL) was added, and the reaction solution was washed with water (100 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 11c (3.0 g, yield: 32%).
[0355] MS m/z (ESI): 458.1 [M+1]
[0356] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 12.87 (s, 1H), 11.03 (s, 1H), 8.05 (s, 1H), 7.92 (s, 1H), 7.27 (dd, 1H), 7.22 (s, 1H), 7.15 (dd, 1H), 7.00 (s, 1H), 6.87-6.82 (m, 1H), 3.71 (s, 3H).
Example 12
5-Chloro-2-(2-cyclopropoxy-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 12
[0357] ##STR00111##
Step 1
1-Bromo-2-cyclopropoxy-4-fluorobenzene 12b
[0358] 2-Bromo-5-fluorophenol 12a (2 g, 10.5 mmol, Accela ChemBio (Shanghai) Inc.), cyclopropyl bromide (5 g, 41.3 mmol, Shanghai Titan Scientific Co., Ltd.), cesium carbonate (7 g, 21.5 mmol, Accela ChemBio (Shanghai) Inc.) and potassium iodide (180 mg, 1.1 mmol) were added to N,N-dimethylformamide (10 mL). The reaction solution was reacted in a microwave reactor at 130° C. for 1.5 hours, and then cooled to room temperature. Ethyl acetate (20 mL) was added, and the reaction solution was washed with water (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 12b (3.0 g, yield: 70%).
Step 2
2-Cyclopropoxy-4-fluorophenol 12c
[0359] Compound 12b (1.85 g, 8 mmol) and triisopropyl borate (1.96 g, 10.4 mmol, Shanghai Titan Scientific Co., Ltd.) were added to 20 mL of tetrahydrofuran. The air in the reaction flask was replaced with argon. The reaction solution was cooled to −78° C., then n-butyl lithium (1.6 M, 7.5 mL, 12 mmol) was slowly added dropwise within 20 minutes. The reaction solution was naturally warmed up to room temperature and stirred overnight. The reaction solution was cooled to 0° C. in an ice bath. 50 mL of methanol was added, and hydrogen peroxide (30 wt %, 11 mL) and 10% sodium hydroxide solution (50 mL) were added dropwise. After completion of the addition, 400 mL of saturated sodium chloride solution was added, and the reaction solution was extract with ethyl acetate (200 mL×3). The organic phase was washed with saturated sodium bicarbonate solution (150 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 12b (1.0 g, yield: 74%).
Step 3
5-Chloro-2-(2-cyclopropoxy-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 12
[0360] Compound 11b (700 mg, 2.1 mmol), compound 12c (300 mg, 1.78 mmol) and cesium carbonate (700 mg, 2.1 mmol, Accela ChemBio (Shanghai) Inc.) were added to 7 mL of N-methylpyrrolidone. The reaction solution was reacted at 80° C. for 1 hour, and then cooled to room temperature. Ethyl acetate (20 mL) was added, and the reaction solution was washed with water (10 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 12 (300 mg, yield: 30%).
[0361] MS m/z (ESI): 484.0 [M+1]
[0362] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 12.84 (s, 1H), 10.99 (s, 1H), 8.01 (s, 1H), 7.87 (s, 1H), 7.30-7.19 (m, 3H), 6.96 (s, 1H), 6.85-6.83 (m, 1H), 3.90-3.88 (m, 3H), 0.74-0.70 (m, 1H), 0.40-0.38 (m, 1H).
Example 13
5-Chloro-2-(4-fluoro-2-methylbenzyl)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 13
[0363] ##STR00112## ##STR00113##
Step 1
(4-Fluoro-2-methylphenyl)magnesium bromide 13b
[0364] A polished magnesium bar (760 mg, 31.7 mmol, Shanghai Sinopharm Chemical Reagent Co., Ltd.) was cut into small pieces and add to tetrahydrofuran (80 mL) under an argon atmosphere. Trimethylchlorosilane (345 mg, 3.17 mmol, Accela ChemBio (Shanghai) Inc.) was added dropwise at room temperature, followed by the addition of 1-bromo-4-fluoro-2-methylbenzene 13a (1.5 g, 7.9 mmol, Accela ChemBio (Shanghai) Inc.). After the reaction was initiated by heating, additional compound 13a (4.5 g, 23.7 mmol, Accela ChemBio (Shanghai) Inc.) was added. The reaction solution was heated to 45° C. and reacted for 1 hour. The magnesium bar disappeared completely, and a grey homogeneous liquid was obtained, i.e., a solution of the title compound 13b (0.4 M, 80 mL), which was used directly in the next step without purification.
Step 2
2-Bromo-4-chloro-5-(trifluoromethyl)benzaldehyde 13d
[0365] Tetrahydrofuran (100 mL) and lithium hexamethyldisilazide (1 M, 120 mL, 120 mmol, Titan Scientific Co., Ltd.) were cooled to −78° C. under an argon atmosphere. 4-Bromo-2-chloro-1-(trifluoromethyl)benzene 13c (25 g, 96.36 mmol, Accela ChemBio (Shanghai) Inc.) was added dropwise, and the reaction solution was kept at this low temperature and reacted for 2 hours. N,N-Dimethylformamide (14.1 g, 192.9 mmol, J&K Scientific Ltd.) was added dropwise, and the reaction solution was gradually warmed up to room temperature and reacted for 16 hours. Water was added, and the reaction solution was extracted with ethyl acetate (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 13d (3.14 g, yield: 28%).
[0366] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 10.32 (s, 1H), 8.23 (s, 1H), 7.87 (s, 1H).
Step 3
(2-Bromo-4-chloro-5-(trifluoromethyl)phenyl)(4-fluoro-2-methylphenyl)methanol 13e
[0367] Compound 13d (900 mg, 3.13 mmol) was dissolved in tetrahydrofuran (10 mL). A freshly prepared solution of compound 13b (7.97 mmol, 19.92 mL) was added dropwise, and the reaction solution was reacted at room temperature for 1 hour. Saturated ammonium chloride solution was added, and the reaction solution was extracted with ethyl acetate (10 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 13e (900 mg, yield: 72%).
[0368] 1HNMR (400 MHz, CD.sub.3OD) δ 8.04 (s, 1H), 7.93 (s, 1H), 7.0-7.03 (m, 1H), 6.87-6.90 (m, 2H), 6.17 (s, 1H), 2.5 (s, 3H).
Step 4
1-Bromo-5-chloro-2-(4-fluoro-2-methylbenzyl)-4-(trifluoromethyl)benzene 13f
[0369] Compound 13e (4 g, 10.1 mmol) was dissolved in dichloromethane (50 mL). The resulting solution was cooled to 0° C., trifluoroacetic acid (10 mL, Titan Scientific Co., Ltd.) was added, and then triethylsilane (6 mL, Accela ChemBio (Shanghai) Inc.) was added dropwise. The reaction solution was reacted at 0° C. for 1 hour. Water was added, and the reaction solution was extracted with ethyl acetate (10 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 13f (3.2 g, yield: 83%).
Step 5
Methyl 5-chloro-2-(4-fluoro-2-methylbenzyl)-4-(trifluoromethyl)benzoate 13g
[0370] Compound 13f (3.3 g, 8.64 mmol) was dissolved in methanol (60 mL). Palladium acetate (388.31 mg, 1.73 mmol, J&K Scientific Ltd.), 1,1′-bis(diphenyphosphino)ferrocene (960 mg, 1.73 mmol, Accela ChemBio (Shanghai) Inc.) and triethylamine (2.63 g, 25.94 mmol, Shanghai Sinopharm Chemical Reagent Co., Ltd.) were added. The reaction system was connected to a carbon monoxide balloon, and reacted at 60° C. for 16 hours. The reaction solution was filtered through diatomaceous earth. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 13g (2.2 g, yield: 71%).
[0371] MS m/z (ESI): 359.1[M−1]
Step 6
5-Chloro-2-(4-fluoro-2-methylbenzyl)-4-(trifluoromethyl)benzoic acid 13h
[0372] Compound 13g (2.2 g, 6.1 mmol) was dissolved in methanol (40 mL) and water (20 mL), followed by the addition of sodium hydroxide solution (5 M, 6 mL, 30 mmol). The reaction solution was warmed up to 40° C. and reacted for 3 hours. The reaction solution was cooled, adjusted to pH 2 with 4 M hydrochloric acid, and extracted with dichloromethane (10 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain the crude title compound 13h (2.1 g), which was used directly in the next step without purification.
[0373] MS m/z (ESI): 345.1[M−1]
Step 7
5-Chloro-N-(6-chloropyridazin-4-yl)-2-(4-fluoro-2-methylbenzyl)-4-(trifluoromethyl)benzamide 13i
[0374] Compound 13h (300 mg, 0.87 mmol) was dissolved in dichloromethane (15 mL), followed by the addition of one drop of N,N-dimethylformamide. Thionyl chloride (2 mL, Shanghai Sinopharm Chemical Reagent Co., Ltd.) was added dropwise in an ice bath. The reaction solution was reacted at room temperature overnight, and then concentrated under reduced pressure. The resulting residue was dissolved in pyridine (2 mL), followed by the addition of 4-amino-6-chloropyridazine (168 mg, 1.3 mmol, Accela ChemBio (Shanghai) Inc.). The reaction solution was reacted at room temperature overnight, and then concentrated under reduced pressure. Water (20 mL) was added, and the resulting solution was extracted with dichloromethane (10 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain the crude compound 13i (260 mg), which was used directly in the next step without purification.
[0375] MS m/z (ESI): 458.1[M+1]
Step 8
5-Chloro-2-(4-fluoro-2-methylbenzyl)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 13
[0376] Compound 13i (260 mg, 0.57 mmol) was dissolved in acetic acid (5 mL), followed by the addition of potassium acetate (112 mg, 1.14 mmol), and the reaction solution was reacted at 130° C. overnight. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by preparative high performance liquid chromatography (Waters 2767-SQ Detecor2, eluent system: ammonium bicarbonate, water, acetonitrile) to obtain the title compound 13 (10 mg, yield of two steps: 2.6%).
[0377] MS m/z (ESI): 440.1 [M+1]
[0378] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.90-7.91 (m, 1H), 7.81 (m, 1H), 7.59 (m, 1H), 7.39 (m, 1H), 6.95-6.97 (m, 1H), 6.85-6.88 (m, 1H), 6.78-6.79 (m, 1H), 4.63 (m, 1H), 4.19 (s, 2H), 2.20 (s, 3H).
Example 14
5-Chloro-2-(2-fluoro-4-(trifluoromethoxy)phenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 14
[0379] ##STR00114##
Step 1
2-Fluoro-1-nitro-4-(trifluoromethoxy)benzene 14b
[0380] 1-Fluoro-3-(trifluoromethoxy)benzene 14a (5 g, 27.76 mmol, Accela ChemBio (Shanghai) Inc.) was dissolved in sulfuric acid (20 mL), and the resulting solution was cooled in an ice bath. Potassium nitrate (7 g, 69.2 mmol) was added in batches, and the reaction solution was naturally warmed up to room temperature and reacted overnight. The reaction solution was poured into ice water, stirred for 30 minutes, extracted with ethyl acetate three times, dried over sodium sulfate, and concentrated to dryness by rotary evaporation to obtain a crude mixture (5.8 g) containing compound 14b.
Step 2
2-Fluoro-4-(trifluoromethoxy)aniline 14c
[0381] The crude mixture (5.8 g, 25.7 mmol) containing compound 14b was dissolved in methanol (80 mL). The resulting solution was purged with nitrogen, and Pd/C catalyst (1.54 g, 14.47 mmol) was added. The resulting solution was purged with hydrogen three times, and the hydrogenation reaction was carried out at room temperature overnight. The reaction solution was filtered, and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography with eluent system A to obtain a crude mixture (3.5 g) containing the title compound 14c.
Step 3
1-Bromo-2-fluoro-4-(trifluoromethoxy)benzene 14d
[0382] The mixture (3.4 g, 17.4 mmol) containing compound 14c was dissolved in acetonitrile (40 mL), followed by the addition of copper bromide (4.67 g, 20.9 mmol, Accela ChemBio (Shanghai) Inc.) and tert-butyl nitrite (2.16 g, 20.9 mmol). The reaction solution was reacted at 60° C. for 0.5 hour, and then filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain a mixture (1.3 g) containing the title compound 14d.
Step 4
2-(2-Fluoro-4-(trifluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane 14e
[0383] The mixture (3 g, 11.6 mmol) containing compound 14d was dissolved in 1,4-dioxane (50 mL), followed by the addition of pinacol diborate (4.4 g, 17.3 mmol, Accela ChemBio (Shanghai) Inc.), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (424 mg, 0.58 mmol) and potassium acetate (3.41 g, 34.7 mmol). The reaction solution was purged with argon three times, and reacted at 100° C. overnight. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with eluent system A to obtain a mixture (950 mg) containing the title compound 14e.
Step 5
2-Fluoro-4-(trifluoromethoxy)phenol 14f
[0384] The mixture (950 mg, 3.1 mmol) containing compound 14e was dissolved in tetrahydrofuran (20 mL), followed by the addition of sodium hydroxide solution (621 mg, 15.5 mmol, 5 mL). The resulting solution was cooled in an ice bath. Hydrogen peroxide (3.1 mL) was added dropwise, and the reaction solution was reacted at room temperature overnight. The organic phase was washed with saturated sodium bicarbonate solution (50 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain a mixture (220 mg) containing the title compound 14f.
Step 6
Methyl 5-chloro-2-(2-fluoro-4-(trifluoromethoxy)phenoxy)-4-(trifluoromethyl)benzoate 14g
[0385] Compound 2c (300 mg, 1.2 mmol) and the mixture (229 mg, 1.2 mmol) containing compound 14f were added to N,N-dimethylformamide (5 mL), followed by the addition of cesium carbonate (571 mg, 1.8 mmol). The reaction solution was reacted at 100° C. for 1 hour, cooled and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain a mixture (440 mg) containing the title compound 14g.
Step 7
5-Chloro-2-(2-fluoro-4-(trifluoromethoxy)phenoxy)-4-(trifluoromethyl)benzoic acid 14h
[0386] Compound 14g (400 mg, 0.92 mmol) was dissolved in tetrahydrofuran (5 mL), followed by the addition of lithium hydroxide solution (194 mg, 4.6 mmol, 1 mL). The reaction solution was reacted at room temperature for 4 hours, concentrated under reduced pressure, followed by the addition of water (5 mL). The resulting solution was adjusted to pH=4 with diluted hydrochloric acid, and extracted with ethyl acetate three times (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a mixture (340 mg) containing the title compound 14h, which was used directly in the next step.
Step 8
5-Chloro-2-(2-fluoro-4-(trifluoromethoxy)phenoxy)-4-(trifluoromethyl)benzoyl chloride 14i
[0387] Compound 14h (80 mg, 0.19 mmol) was dissolved in thionyl chloride (2 mL), and the reaction solution was reacted at 80° C. for 2 hours. The reaction solution was concentrated under reduced pressure to obtain a mixture (80 mg) containing the title compound 14i, which was used directly in the next step.
Step 9
5-Chloro-N-(6-chloropyridazin-4-yl)-2-(2-fluoro-4-(trifluoromethoxy)phenoxy)-4-(trifluoromethyl)benzamide 14j
[0388] 4-Amino-6-chloropyridazine (31 mg, 0.24 mmol, Accela ChemBio (Shanghai) Inc.) was dissolved in tetrahydrofuran (3 mL), and the resulting solution was cooled in an ice bath. Sodium hydride (13 mg, 0.34 mmol, purity: 60%) was added, and the reaction solution was reacted for 30 minutes. A solution (2 mL) of compound 14i in tetrahydrofuran was added dropwise, and the reaction solution was reacted at room temperature overnight under an argon atmosphere. Water (10 mL) and ethyl acetate (20 mL) were added, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with eluent system B to obtain a mixture (65 mg) containing the title compound 14j.
Step 10
5-Chloro-2-(2-fluoro-4-(trifluoromethoxy)phenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 14
[0389] The mixture (65 mg, 0.12 mmol) containing compound 14j was dissolved in acetic acid (3 mL), followed by the addition of potassium acetate (60 mg, 0.61 mmol), and the reaction solution was reacted at 130° C. for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by preparative high performance liquid chromatography (Waters 2767-SQ Detecor2, eluent system: ammonium bicarbonate, water, acetonitrile) to obtain the title compound 14 (8 mg, yield: 11%).
[0390] MS m/z (ESI): 512.0 [M+1]
[0391] H NMR (400 MHz, CD.sub.3OD): δ 8.02 (d, 1H), 8.00 (s, 1H), 7.43 (d, 1H), 7.40 (s, 1H), 7.37-7.27 (m, 2H), 7.19 (d, 1H).
Example 15
5-Chloro-2-(4-fluoro-2-(trifluoromethoxy)phenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 15
[0392] ##STR00115## ##STR00116##
Step 1
1-Methoxy-2-(trifluoromethoxy)benzene 15b
[0393] 2-(Trifluoromethoxy)phenol 15a (5 g, 28.1 mmol, Accela ChemBio (Shanghai) Inc.) was dissolved in N,N-dimethylformamide (50 mL), followed by the addition of methyl iodide (4.78 g, 33.7 mmol) and potassium carbonate (7.75 g, 56.1594 mmol). The reaction solution was reacted at 80° C. for 1 hour. Ethyl acetate and water were added to the reaction solution, and the aqueous phase was extracted with ethyl acetate three times. The organic phases were combined, washed with water four times, dried over sodium sulfate, and concentrated to dryness by rotary evaporation to obtain the title compound 15b (5.1 g, yield: 94%).
Step 2
1-Methoxy-4-nitro-2-(trifluoromethoxy)benzene 15c
[0394] Sodium nitrate (2.26 g, 26.6 mmol) was dissolved in trifluoroacetic acid (50 mL). The resulting solution was cooled in an ice bath, and a solution (10 mL) of compound 15b (5.1 g, 26.5 mmol) in trifluoroacetic acid was added. The reaction solution was reacted in the ice bath for 1 hour, and at room temperature for 3 hours. Water (50 mL) and ethyl acetate (50 mL) were added to the reaction solution. The organic phase was washed with saturated sodium bicarbonate solution to be neutral, dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the title compound 15c (5.3 g, yield: 84%).
Step 3
4-Methoxy-3-(trifluoromethoxy)aniline 15d
[0395] Compound 15c (5.3 g, 22.3 mmol) was dissolved in methanol (100 mL), followed by the addition of concentrated hydrochloric acid (0.5 mL), and Pd/C catalyst (500 mg). The resulting solution was purged with hydrogen three times, and the hydrogenation reaction was carried out at room temperature for 4 hours. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 15d (3.1 g, yield: 67%).
Step 4
4-Fluoro-1-methoxy-2-(trifluoromethoxy)benzene 15e
[0396] Compound 15d (800 mg, 3.9 mmol) was added to a mixed solution of water (15 mL), fluoroboric acid (1.51 g, 7.0 mmol, purity: 40%) and hydrochloric acid (687 mg, 7.0 mmol). The resulting solution was cooled in an ice bath. Sodium nitrite solution (280 mg, 4.06 mmol, 2 mL) was added dropwise, and the reaction solution was reacted in the ice bath for 2 hours. The reaction solution was filtered, and the filter cake was dried. The solid was heated to 130° C. under a nitrogen atmosphere, gradually melted and turned red, and reacted for 1 hour. After the reaction system was cooled, the resulting product was dissolved in dichloromethane, dried over sodium sulfate, and concentrated under reduced pressure to obtain the title compound 15e (600 mg, yield: 74%).
Step 5
4-Fluoro-2-(trifluoromethoxy)phenol 15f
[0397] Compound 15e (300 mg, 1.4 mmol) was dissolved in dichloromethane (5 mL), and the resulting solution was cooled in an ice bath. Boron tribromide solution (1 M, 7 mL) was added, and the reaction solution was naturally warmed up to room temperature and reacted overnight. The reaction solution was added dropwise to methanol, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 15f (100 mg, yield: 36%).
Step 6
5-Chloro-2-(4-fluoro-2-(trifluoromethoxy)phenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 15
[0398] Compound 15f (300 mg, 0.89 mmol) was added to N-methylpyrrolidone (3 mL), followed by the addition of compound 11b (140 mg, 0.71 mmol) and cesium carbonate (291 mg, 0.9 mmol). The reaction solution was reacted at 80° C. for 1 hour. Water (10 mL) and ethyl acetate (20 mL) were added to the reaction solution, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by preparative high performance liquid chromatography (Waters 2767-SQ Detecor2, eluent system: ammonium bicarbonate, water, acetonitrile) to obtain the title compound 15 (40 mg, yield: 11%).
[0399] MS m/z (ESI): 512.0 [M+1]
[0400] H NMR (400 MHz, CD.sub.3OD): δ 8.03 (d, 1H), 8.01 (s, 1H), 7.46 (d, 1H), 7.37-7.35 (m, 1H), 7.32-7.22 (m, 3H).
Example 16
2-(4-Fluoro-2-methoxyphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-5-(trifluoromethyl)benzamide 16
[0401] ##STR00117##
[0402] In accordance with the synthetic route in Example 2, the starting compound 5-chloro-2-fluoro-4-(trifluoromethyl)benzoic acid in Step 2 was replaced with compound 2-fluoro-5-(trifluoromethyl)benzoic acid, and the starting compound 4-fluoro-2-methyl-phenol in Step 3 was replaced with 4-fluoro-2-methoxy-phenol, accordingly, the title compound 16 (27 mg) was prepared.
[0403] MS m/z (ESI): 424.1[M+1]
[0404] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 312.81 (s, 1H), 10.90 (s, 1H), 7.95 (d, 2H), 7.76 (d, 1H), 7.21-7.33 (m, 2H), 7.15 (d, 1H), 6.72-6.90 (m, 2H), 3.71 (s, 3H).
Example 17
5-Chloro-2-((4-fluoro-2-methoxyphenyl)thio)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 17
[0405] ##STR00118##
[0406] In accordance with the synthetic route in Example 2, the starting compound 2-methyl-4-fluorophenol in Step 3 was replaced with 2-methoxybenzenethiol, accordingly, the title compound 17 (35 mg) was prepared.
[0407] MS m/z (ESI): 474.0 [M+1]
[0408] 1H NMR (400 MHz, DMSO-d.sub.6) δ 12.90 (s, 1H), 11.06 (s, 1H), 8.12 (s, 1H), 7.97 (s, 1H), 7.59-7.55 (m, 1H), 7.25 (s, 1H), 7.16-7.13 (m, 2H), 6.93-6.89 (m, 1H), 3.74 (s, 3H).
Example 18
4-Chloro-5-fluoro-2-(4-fluoro-2-methoxyphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide 18
[0409] ##STR00119##
[0410] In accordance with the synthetic route in Example 1, the starting compound 4,5-dichloro-2-fluorobenzoic acid in Step 1 was replaced with compound 4-chloro-2,5-difluorobenzoic acid, accordingly, the title compound 18 (22 mg) was prepared.
[0411] MS m/z (ESI): 408.0 [M+1]
[0412] 1H NMR (400 MHz, DMSO-d.sub.6) 312.81 (s, 1H), 10.83 (s, 1H), 7.90-7.89 (d, 1H), 7.77-7.75 (d, 1H), 7.19-7.16 (m, 2H), 7.10-7.06 (m, 1H), 6.90-6.81 (m, 1H), 6.81-6.77 (m, 1H), 3.71 (s, 3H).
Example 19
4-Chloro-5-fluoro-2-(4-fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide 19
[0413] ##STR00120##
[0414] In accordance with the synthetic route in Example 1, the starting compound 4,5-dichloro-2-fluorobenzoic acid in Step 1 was replaced with compound 4-chloro-2,5-difluorobenzoic acid, and the starting compound 2-methoxy-4-fluorophenol in Step 3 was replaced with 2-methyl-4-fluorophenol, accordingly, the title compound 19 (9 mg) was prepared as a white solid.
[0415] MS m/z (ESI): 392.1 [M+1]
[0416] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 312.81 (s, 1H), 10.89 (s, 1H), 7.87-7.86 (d, 1H), 7.82-7.80 (d, 1H), 7.18-7.14 (m, 2H), 7.04-7.03 (m, 2H), 7.00-6.93 (m, 1H), 2.14 (s, 3H).
Example 20
4-Chloro-2-(4-fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide 20
[0417] ##STR00121##
[0418] In accordance with the synthetic route in Example 2, the starting compound 5-chloro-2-fluoro-4-(trifluoromethyl)benzoic acid in Step 2 was replaced with compound 4-chloro-2-fluorobenzoic acid, accordingly, the title compound 20 (40 mg) was prepared.
[0419] MS m/z (ESI): 374.1 [M+1]
[0420] 1H NMR (400 MHz, DMSO-d.sub.6) 312.79 (s, 1H), 10.83 (s, 1H), 7.89 (s, 1H), 7.67 (d, 1H), 7.30 (d, 1H), 7.14-7.22 (m, 2H), 7.00-7.12 (m, 2H), 6.74 (s, 1H), 2.13 (s, 3H).
Example 21
4-Chloro-2-(4-fluoro-2-methoxyphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide
[0421] ##STR00122##
[0422] In accordance with the synthetic route in Example 2, the starting compound 5-chloro-2-fluoro-4-(trifluoromethyl)benzoic acid in Step 2 was replaced with compound 4-chloro-2-fluorobenzoic acid, and the starting compound 2-methyl-4-fluorophenol in Step 3 was replaced with compound 2-methoxy-4-fluorophenol, accordingly, the title compound 21 (40 mg) was prepared.
[0423] MS m/z (ESI): 390.1 [M+1]
[0424] 1H NMR (400 MHz, DMSO-d.sub.6) 312.79 (s, 1H), 10.75 (s, 1H), 7.92 (s, 1H), 7.63 (d, 1H), 7.18-7.30 (m, 3H), 7.10 (dd, 1H), 6.78-6.88 (m, 1H), 6.64 (s, 1H), 3.72 (s, 3H).
Example 22
2-(4-Fluoro-2-methoxyphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-5-(trifluoromethoxy)benzamide 22
[0425] ##STR00123##
[0426] In accordance with the synthetic route in Example 1, the starting compound 4,5-dichloro-2-fluorobenzoic acid in Step 1 was replaced with compound 2-fluoro-5-(trifluoromethoxy)benzoic acid, accordingly, the title compound 22 (5 mg) was prepared.
[0427] MS m/z (ESI): 440.1 [M+1]
[0428] .sup.1H NMR (400 MHz, CD.sub.3OD): δ 8.05 (d, 1H), 7.74 (d, 1H), 7.54 (d, 1H), 7.41-7.38 (m, 1H), 7.26 (q, 1H), 7.03-7.00 (m, 1H), 6.89 (d, 1H), 6.81-6.77 (m, 1H), 3.81 (s, 3H).
Example 23
5-Chloro-2-(4-fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide 23
[0429] ##STR00124##
[0430] In accordance with the synthetic route in Example 1, the starting compound 4,5-dichloro-2-fluorobenzoic acid in Step 1 was replaced with compound 5-chloro-2-fluorobenzoic acid, accordingly, the title compound 23 (14 mg) was prepared.
[0431] MS m/z (ESI): 374.1 [M+1]
[0432] .sup.1H NMR (400 MHz, CD.sub.3OD): δ 8.05 (d, 1H), 7.76 (d, 1H), 7.49-7.46 (m, 2H), 7.07 (d, 1H), 7.01-6.98 (m, 2H), 6.78 (d, 1H), 2.22 (s, 3H).
Example 24
2-(4-Fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-5-(trifluoromethoxy)benzamide 24
[0433] ##STR00125##
[0434] In accordance with the synthetic route in Example 1, the starting compound 4,5-dichloro-2-fluorobenzoic acid in Step 1 was replaced with compound 2-fluoro-5-(trifluoromethoxy)benzoic acid, accordingly, the title compound 24 (7 mg) was prepared.
[0435] MS m/z (ESI): 424.1 [M+1]
[0436] .sup.1H NMR (400 MHz, CD.sub.3OD): δ 8.05 (d, 1H), 7.71 (d, 1H), 7.50 (d, 1H), 7.44-7.41 (m, 1H), 7.11-6.97 (m, 3H), 6.87 (d, 1H), 2.23 (s, 3H).
Example 25
5-Fluoro-2-(4-fluoro-2-methoxyphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide 25
[0437] ##STR00126##
[0438] In accordance with the synthetic route in Example 1, the starting compound 4,5-dichloro-2-fluorobenzoic acid in Step 1 was replaced with compound 2,5-difluorobenzoic acid, accordingly, the title compound 25 (20 mg) was prepared.
[0439] MS m/z (ESI): 374.0 [M+1]
[0440] 1H NMR (400 MHz, DMSO-d.sub.6) δ 12.78 (s, 1H), 10.77 (s, 1H), 7.91 (s, 1H), 7.45-7.55 (m, 1H), 7.00-7.32 (m, 4H), 6.65-6.85 (m, 2H), 3.70 (s, 3H).
Example 26
5-Fluoro-2-(4-fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide 26
[0441] ##STR00127##
[0442] In accordance with the synthetic route in Example 1, the starting compound 4,5-dichloro-2-fluorobenzoic acid in Step 1 was replaced with compound 2,5-difluorobenzoic acid, and the starting compound 2-methoxy-4-fluorophenol in Step 3 was replaced with 2-methyl-4-fluorophenol, accordingly, the title compound 26 (20 mg) was prepared.
[0443] MS m/z (ESI): 358.1 [M+1]
[0444] 1H NMR (400 MHz, DMSO-d.sub.6) δ 12.79 (s, 1H), 10.85 (s, 1H), 7.88 (s, 1H), 7.50-7.60 (m, 1H), 7.30-7.40 (m, 1H), 7.08-7.19 (m, 2H), 6.95-7.05 (m, 1H), 6.80-6.95 (m, 2H), 2.14 (s, 3H).
Example 27
4-Chloro-2-(4-fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-5-(trifluoromethyl)benzamide 27
[0445] ##STR00128## ##STR00129##
Step 1
1-Chloro-5-fluoro-4-nitro-2-(trifluoromethyl)benzene 27b
[0446] 2-Chloro-4-fluoro-1-(trifluoromethyl)benzene 27a (3 g, 15.1 mmol, Adamas Reagent, Co., Ltd.) was dissolved in sulfuric acid (30 mL), and the resulting solution was cooled to −10° C. Potassium nitrate (1.83 g, 18.1 mmol) was added in batches, and the reaction solution was reacted at −10° C. for 1 hour, and at room temperature overnight. The reaction solution was poured into ice and extract with ethyl acetate (50 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain the title compound 27b (3.46 g, yield: 94%).
Step 2
1-Chloro-5-(4-fluoro-2-methylphenoxy)-4-nitro-2-(trifluoromethyl)benzene 27c
[0447] Compound 27b (1 g, 4.1 mmol) was dissolved in N,N-dimethylformamide (10 mL), followed by the addition of 4-fluoro-2-methylphenol (518 mg, 4.1 mmol) and potassium phosphate (2.61 g, 12.3 mmol). The reaction solution was reacted at 80° C. for 1 hour. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 27c (1.27 g, yield: 88%).
Step 3
4-Chloro-2-(4-fluoro-2-methylphenoxy)-5-(trifluoromethyl)aniline 27d
[0448] Compound 27c (1.2 g, 3.4 mmol) was dissolved in ethanol (20 mL) and water (10 mL), followed by the addition of reduced iron powder (1.15 g, 20.6 mmol) and ammonium chloride (1.10 g, 20.6 mmol). The reaction solution was reacted at 80° C. for 3 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 27d (900 mg, yield: 82%).
Step 4
1-Bromo-4-chloro-2-(4-fluoro-2-methylphenoxy)-5-(trifluoromethyl)benzene 27e
[0449] Compound 27d (1 g, 3.1 mmol) was dissolved in acetonitrile (10 mL), followed by the addition of copper bromide (839 mg, 3.7 mmol, Adamas Reagent, Co., Ltd.) and tert-butyl nitrite (387 mg, 3.7 mmol). The reaction solution was refluxed for 2 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound 27e (1 g, yield: 83%).
Step 5
Methyl 4-chloro-2-(4-fluoro-2-methylphenoxy)-5-(trifluoromethyl)benzoate 27f
[0450] Compound 27e (400 mg, 1.04 mmol) was dissolved in methanol (10 mL), followed by the addition of palladium acetate (47 mg, 0.21 mmol), 1,1′-bis(diphenyphosphino)ferrocene (116 mg, 0.21 mmol) and triethylamine (317 mg, 3.1 mmol). The resulting solution was purged with carbon monoxide three times. The reaction solution was reacted at 70° C. overnight under a carbon monoxide atmosphere. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 27f (400 mg, yield: 42%).
Step 6
4-Chloro-2-(4-fluoro-2-methylphenoxy)-5-(trifluoromethyl)benzoic acid 27g
[0451] Compound 27f (480 mg, 1.3 mmol) was dissolved in tetrahydrofuran (5 mL) and water (1 mL), followed by the addition of lithium hydroxide monohydrate (170 mg, 4.0 mmol). The reaction solution was reacted at room temperature overnight. The reaction solution was concentrated under reduced pressure. The resulting residue was dissolved by adding a small amount of water, and the pH was adjusted to 4 with dilute hydrochloric acid. The resulting solution was extracted with ethyl acetate (20 mL×3), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the title compound 27g (430 mg, yield: 93%).
Step 7
4-Chloro-2-(4-fluoro-2-methylphenoxy)-5-(trifluoromethyl)benzoyl chloride 27h
[0452] Compound 27g (200 mg, 0.57 mmol) was added to thionyl chloride (2 mL), and the reaction solution was reacted at 80° C. for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the crude title compound 27h (210 mg), which was used directly in the next step.
Step 8
4-Chloro-N-(6-chloropyridin-4-yl)-2-(4-fluoro-2-methylphenoxy)-5-(trifluoromethyl)benzamide 27i
[0453] Compound 27h (230 mg, 0.73 mmol) was dissolved in pyridine (3 mL), followed by the addition of 4-amino-6-chloropyridazine (95 mg, 0.73 mmol) and 4-dimethylaminopyridine (10 mg, 0.08 mmol). The reaction solution was reacted at room temperature overnight. The reaction solution was concentrated, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 27i (60 mg, yield: 20%).
Step 9
4-Chloro-2-(4-fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-5-(trifluoromethyl)benzamide 27
[0454] Compound 27i (60 mg, 0.13 mmol) was dissolved in acetic acid (2 mL), followed by the addition of potassium acetate (25 mg, 0.25 mmol). The reaction solution was reacted at 130° C. for 4 hours, and then concentrated under reduced pressure. The resulting residue was purified by preparative high performance liquid chromatography (Waters 2767-SQ Detecor2, eluent system: ammonium bicarbonate, water, acetonitrile) to obtain the title compound 27 (14 mg, yield: 24%).
[0455] MS m/z (ESI): 442.1 [M+1]
[0456] 1H NMR (400 MHz, CD.sub.3OD): δ 8.18 (s, 1H), 8.06 (d, 1H), 7.52 (d, 1H), 7.18-7.15 (m, 2H), 7.09-7.06 (m, 1H), 6.87 (s, 1H), 2.21 (s, 3H).
Example 28
5-Chloro-2-(4-fluoro-2-methylphenoxy)-4-methyl-N-(6-oxo-1,6-dihydropyridazin-4-yl)benzamide 28
[0457] ##STR00130##
[0458] In accordance with the synthetic route in Example 2, the starting compound 5-chloro-2-fluoro-4-(trifluoromethyl)benzoic acid in Step 2 was replaced with compound 5-chloro-2-fluoro-4-methylbenzoic acid, accordingly, the title compound 28 (3 mg) was prepared.
[0459] MS m/z (ESI): 388.2 [M+1]
[0460] .sup.1H NMR (400 MHz, CD.sub.3OD): δ 8.03 (d, 1H), 7.79 (s, 1H), 7.48 (d, 1H), 7.09-7.06 (m, 1H), 7.00-6.96 (s, 2H), 6.72 (s, 1H), 2.33 (s, 3H), 2.22 (s, 3H).
Example 29
2-(4-Fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4,6-bis(trifluoromethyl)benzamide 29
[0461] ##STR00131##
[0462] In accordance with the synthetic route in Example 1, the starting compound 5-chloro-2-fluoro-4-(trifluoromethyl)benzoic acid in Step 1 was replaced with compound 2-fluoro-4,6-bis(trifluoromethyl)benzoic acid, accordingly, the title compound 29 (2.4 mg) was prepared.
[0463] MS m/z (ESI): 476.0 [M+1]
[0464] 1H NMR (400 MHz, CD.sub.3OD) δ 8.02 (s, 1H), 7.82 (s, 1H), 7.50 (s, 1H), 7.00-7.20 (m, 4H), 2.19 (s, 3H).
Example 30
5-Chloro-N-(6-oxo-1,6-dihydropyridazin-4-yl)-2-(o-tolyloxy)-4-(trifluoromethyl)benzamide 30
[0465] ##STR00132##
[0466] In accordance with the synthetic route in Example 2, the starting compound 2-methyl-4-fluorophenol in Step 3 was replaced with 2-methylphenol, accordingly, the title compound 30 (75 mg) was prepared.
[0467] MS m/z (ESI): 424.1 [M+1]
[0468] 1H NMR (400 MHz, DMSO-d.sub.6) 312.87 (s, 1H), 11.08 (s, 1H), 8.11 (s, 1H), 7.88 (d, 1H), 7.34-7.32 (m, 1H), 7.27-7.23 (m, 1H), 7.17-7.13 (m, 3H), 7.02-7.00 (m, 1H), 2.16 (s, 3H).
Example 31
5-Chloro-2-(2-cyclopropyl-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 31
[0469] ##STR00133##
Step 1
2-Cyclopropyl-4-fluorophenol 31b
[0470] 2-Bromo-4-fluorophenol 31a (1.77 g, 9.26 mmol, Shanghai Bide Pharmatech Ltd.), tripotassium phosphate (6.89 g, 32.46 mmol), tricyclohexylphosphine (260 mg, 0.93 mmol) and cyclopropylboronic acid (1.20 g, 13.97 mmol, Shanghai Bide Pharmatech Ltd.) were added to a mixed solution of toluene (40 mL)/water (2 mL). The resulting solution was purged with argon three times. Palladium acetate (105 mg, 0.46 mmol) was added, and the reaction solution was purged with argon three times and reacted at 100° C. overnight. The reaction solution was cooled, followed by the addition of ethyl acetate (50 mL), and washed with water (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain the crude title compound 31b (1.41 g).
[0471] MS m/z (ESI): 151.1 [M−1]
Step 2
5-Chloro-2-(2-cyclopropyl-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 31
[0472] Compound 11b (1.03 g, 3.06 mmol), compound 31b (0.47 g, 3.08 mmol) and cesium carbonate (1.01 g, 3.09 mmol) were added to N-methylpyrrolidone (10 mL). The reaction solution was reacted at 60° C. overnight. The reaction solution was cooled, followed by the addition of ethyl acetate (150 mL), and washed with water (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative high performance liquid chromatography (Waters 2767-SQ Detecor2, eluent system: ammonium bicarbonate, water, acetonitrile) to obtain the title compound 31 (350 mg, yield: 24%).
[0473] MS m/z (ESI): 468.0 [M+1]
[0474] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 12.87 (s, 1H), 11.08 (s, 1H), 8.10 (s, 1H), 7.90 (d, 1H), 7.21 (s, 1H), 7.14 (dd, 1H), 7.08-7.03 (m, 2H), 6.87 (dd, 1H), 2.01-1.94 (m, 1H), 0.87-0.83 (m, 2H), 0.71-0.67 (m, 2H).
Example 32
5-Chloro-2-(2-ethoxy-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 32
[0475] ##STR00134##
[0476] In accordance with the synthetic route in Example 12, the starting compound cyclopropyl bromide in Step 1 was replaced with compound iodoethane to react at room temperature, accordingly, the title compound 32 (200 mg) was prepared.
[0477] MS m/z (ESI): 472.1 [M+1]
[0478] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 312.83 (s, 1H), 10.98 (s, 1H), 8.02 (s, 1H), 7.89 (s, 1H), 7.00-7.28 (m, 4H), 6.73-6.83 (m, 1H), 3.99 (q, 2H), 1.07 (t, 3H).
Example 33
5-Bromo-2-(4-fluoro-2-methylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 33
[0479] ##STR00135##
[0480] In accordance with the synthetic route in Example 11, the starting compound 5-chloro-2-fluoro-4-(trifluoromethyl)benzoic acid in Step 1 was replaced with compound 5-bromo-2-fluoro-4-(trifluoromethyl)benzoic acid, accordingly, the title compound 33 (126 mg) was prepared.
[0481] MS m/z (ESI): 486.1 [M+1]
[0482] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 312.84 (s, 1H), 11.03 (s, 1H), 8.19 (s, 1H), 7.86-7.85 (d, 1H), 7.21-7.16 (m, 2H), 7.07-7.06 (d, 1H), 2.13 (s, 3H).
Example 34
2-(4-Fluoro-2-methylphenoxy)-5-methyl-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 34
[0483] ##STR00136##
[0484] In accordance with the synthetic route in Example 11, the starting compound 5-chloro-2-fluoro-4-(trifluoromethyl)benzoic acid in Step 1 was replaced with compound 5-methyl-2-fluoro-4-(trifluoromethyl)benzoic acid, and the starting compound 2-methoxy-4-fluorophenol in Step 3 was replaced with 2-methyl-4-fluorophenol, accordingly, the title compound 34 (15 mg) was prepared.
[0485] MS m/z (ESI): 422.0 [M+1]
[0486] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.83 (s, 1H), 11.97 (s, 1H), 7.89-7.88 (d, 1H), 7.74 (d, 1H), 7.19-7.18 (m, 2H), 7.08-7.04 (m, 1H), 7.00-6.97 (m, 2H), 2.44 (s, 3H), 2.15 (s, 3H).
Example 35
5-Chloro-2-(4-fluoro-2-(methoxy-d.SUB.3.)phenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 35
[0487] ##STR00137##
Step 1
1-Bromo-4-fluoro-2-(methoxy-d.SUB.3.)benzene 35b
[0488] 2-Bromo-5-fluorophenol 12a (1 g, 5.2 mmol, Accela ChemBio (Shanghai) Inc.), deuterated methyl iodide (911 mg, 6.3 mmol, Sun Chemical Technology (Shanghai) Co., Ltd.) and potassium carbonate (1.45 g, 10.5 mmol) were added to N,N-dimethylformamide (10 mL). The reaction solution was stirred to react for 6 hours. The reaction solution was cooled to room temperature. Ethyl acetate (20 mL) was added, and the reaction solution was washed with water (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 35b (840 mg, yield: 71%).
[0489] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 7.49-7.45 (m, 1H), 6.66-6.57 (m, 2H).
Step 2
4-Fluoro-2-(methoxy-d.SUB.3.)phenol 35c
[0490] Compound 35b (840 mg, 4 mmol) and triisopropyl borate (987 mg, 5.25 mmol, Shanghai Titan Scientific Co., Ltd.) were added to a mixed solution of tetrahydrofuran/toluene (150 mL/30 mL). The air in the reaction flask was replaced with argon. The reaction solution was cooled to −78° C., then n-butyl lithium (1.6 M, 3.8 mL, 6.1 mmol) was slowly added dropwise within 20 minutes. The reaction solution was naturally warmed up to room temperature and stirred overnight. The reaction solution was cooled to 0° C. in an ice bath. Methanol (50 mL) was added, and hydrogen peroxide (30 wt %, 10 mL) and 10% sodium hydroxide solution (40 mL) were added dropwise. The reaction solution was stirred at room temperature for 1 hour. Saturated sodium thiosulfate solution (50 mL) was slowly added dropwise, and the reaction solution was extracted with ethyl acetate (200 mL×3). The organic phase was washed with saturated sodium bicarbonate solution (150 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 35c (570 mg, yield: 97%).
[0491] MS m/z (ESI): 144.0 [M−1]
[0492] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.89 (s, 1H), 6.85-6.82 (m, 1H), 6.76-6.72 (m, 1H), 6.59-6.54 (m, 1H).
Step 3
5-Chloro-2-(4-fluoro-2-(methoxy-d.SUB.3.)phenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 35
[0493] Compound 11b (1 g, 2.98 mmol), compound 35c (433 mg, 2.98 mmol) and cesium carbonate (1.02 g, 3.13 mmol, Accela ChemBio (Shanghai) Inc.) were added to N-methylpyrrolidone (10 mL). The reaction solution was reacted at 80° C. for 3 hours, and cooled to room temperature. Ethyl acetate (20 mL) was added, and the reaction solution was washed with water (10 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 35 (280 mg, yield: 20%).
[0494] MS m/z (ESI): 461.0 [M+1]
[0495] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 12.87 (s, 1H), 11.03 (s, 1H), 8.06 (s, 1H), 7.93 (d, 1H), 7.29-7.23 (m, 2H), 7.16-7.13 (m, 1H), 7.01 (s, 1H), 6.88-6.83 (m, 1H).
Example 36
5-Chloro-2-(2-ethyl-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 36
[0496] ##STR00138##
Step 1
4-Fluoro-2-(1-hydroxyethyl)phenol 36b
[0497] Compound 1-(5-fluoro-2-hydroxyphenyl)ethan-1-one 36a (3 g, 19.5 mmol, Accela ChemBio (Shanghai) Inc.) was dissolved in anhydrous methanol (20 mL). Sodium borohydride (1.1 g, 29.1 mmol) was slowly added in batches, and the reaction solution was reacted at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and ethyl acetate and water were added. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain the crude title compound 36b (3.2 g, yield: 100%).
[0498] MS m/z (ESI): 155.1[M−1]
Step 2
2-Ethyl-4-fluorophenol 36c
[0499] Compound 36b (1.5 g, 9.6 mmol) was dissolved in dichloromethane (15 mL), followed by the addition of trifluoroacetic acid (11 g, 96.5 mmol, 7.2 mL). Triethylsilane (11.2 g, 96.3 mmol, 15.4 mL) was added dropwise, and the reaction solution was reacted at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and dichloromethane and water were added. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain the crude title compound 36c (1.38 g, yield: 100%).
[0500] MS m/z (ESI): 139.1[M−1]
Step 3
5-Chloro-2-(2-ethyl-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 36
[0501] Compound 11b (100 mg, 0.3 mmol), compound 36c (42 mg, 0.3 mmol) and cesium carbonate (100 mg, 0.33 mmol) were added to N-methylpyrrolidone (2 mL). The reaction solution was reacted at 60° C. overnight. The reaction solution was cooled, followed by the addition of ethyl acetate (150 mL), and washed with water (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative high performance liquid chromatography (Waters 2767-SQ Detecor2, eluent system: ammonium bicarbonate, water, acetonitrile) to obtain the title compound 36 (18 mg, yield: 13%).
[0502] MS m/z (ESI): 456.0[M+1]
[0503] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.88 (s, 1H), 11.09 (s, 1H), 8.12 (s, 1H), 7.90-7.90 (d, 1H), 7.25-7.20 (m, 2H), 7.14 (m, 1H), 7.11-7.10 (m, 2H), 2.58-2.54 (m, 2H), 1.10-1.06 (m, 3H).
Example 37
5-Chloro-2-(2-(difluoromethoxy)-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 37
[0504] ##STR00139##
Step 1
2-(Difluoromethoxy)-4-fluoro-1-methoxybenzene 37b
[0505] 5-Fluoro-2-methoxyphenol 37a (1 g, 7.03 mmol, Accela ChemBio (Shanghai) Inc.), sodium difluorochloroacetate (2.68 g, 17.57 mmol, Accela ChemBio (Shanghai) Inc.) and cesium carbonate (4.58 g, 14.05 mmol) were added to a mixed solvent of N,N-dimethylformamide (14 mL)/water (1.5 mL). The reaction solution was reacted at 100° C. overnight. The reaction solution was cooled, followed by the addition of dichloromethane (100 mL), and washed with water (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain the crude title compound 37b (1.35 g).
Step 2
2-(Difluoromethoxy)-4-fluorophenol 37c
[0506] Sodium iodide (5.27 g, 35.15 mmol, Shanghai Sinopharm Chemical Reagent Co., Ltd.) was dissolved in acetonitrile (20 mL), followed by the addition of trimethylchlorosilane (3.82 g, 35.16 mmol, Shanghai Sinopharm Chemical Reagent Co., Ltd.). The reaction solution was reacted at room temperature for 20 minutes. Compound 37b (1.35 g, 7.0263 mmol) was added, and the reaction solution was reacted at 80° C. overnight. The reaction solution was cooled, followed by the addition of water (50 mL), and extracted with ethyl acetate (50 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 37c (540 mg, yield: 43%).
[0507] MS m/z (ESI): 177.1 [M−1]
Step 3
5-Chloro-2-(2-(difluoromethoxy)-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 37
[0508] Compound 11b (1.02 g, 3.03 mmol), compound 37c (0.54 g, 3.03 mmol) and cesium carbonate (0.99 g, 3.13 mmol) were added to N-methylpyrrolidone (10 mL). The reaction solution was reacted at 60° C. overnight. The reaction solution was cooled, followed by the addition of ethyl acetate (150 mL), and washed with water (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative high performance liquid chromatography (Waters 2767-SQ Detecor2, eluent system: ammonium bicarbonate, water, acetonitrile) to obtain the title compound 37 (450 mg, yield: 30%).
[0509] MS m/z (ESI): 493.9 [M+1]
[0510] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 12.86 (s, 1H), 11.06 (s, 1H), 8.11 (s, 1H), 7.89 (d, 1H), 7.38 (dd, 1H), 7.33 (dd, 1H), 7.27 (s, 1H), 7.23 (t, 1H), 7.22-7.17 (m, 2H).
Example 38
5-Chloro-2-(4-fluoro-2-hydroxyphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 38
[0511] ##STR00140##
[0512] Compound 11 (100 mg, 0.22 mmol) was added to dichloromethane (2 mL). The resulting solution was cooled to −78° C., and boron tribromide (1 M, 1.09 mL, Adamas Beta (Shanghai) Reagent, Co., Ltd.) was added dropwise. The reaction solution was warmed up to 0° C. and reacted for 3 hours. Additional boron tribromide (1 M, 1.09 mL) was added at 0° C., and the reaction solution was reacted at room temperature overnight. The reaction solution was concentrated, and 10 mL of methanol was added dropwise to the resulting residue. The resulting solution was stirred well and concentrated. The resulting residue was purified by preparative high performance liquid chromatography (Waters 2767-SQ Detecor2, eluent system: ammonium bicarbonate, water, acetonitrile) to obtain the title compound 38 (50 mg, yield: 51%).
[0513] MS m/z (ESI): 444.1 [M+1]
[0514] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 12.88 (s, 1H), 10.99 (s, 1H), 10.47 (s, 1H), 8.04 (s, 1H), 7.94 (s, 1H), 7.26-7.22 (m, 2H), 6.97 (s, 1H), 6.80 (dd, 1H), 6.75-6.70 (m, 1H).
Example 39
5-Chloro-2-((7-fluoro-2,3-dihydro-1H-inden-4-yl)oxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 39
[0515] ##STR00141##
Step 1
4-Fluorophenyl 3-chloropropanoate 39b
[0516] The starting compound 4-fluorophenol 39a (15 g, 133.8 mmol, Accela ChemBio (Shanghai) Inc.) was dissolved in dichloromethane (80 mL), followed by the addition of pyridine (12 g, 151.7 mmol). 3-Chloropropionyl chloride (18.9 g, 134 mmol, Accela ChemBio (Shanghai) Inc.) was slowly added dropwise under an ice bath. After completion of the addition, the reaction solution was stirred at room temperature for 1 hour. Saturated sodium bicarbonate solution (60 mL) was added, and the reaction solution was extracted with ethyl acetate (80 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 39b (25 g, yield: 86%).
[0517] MS m/z (ESI): 203.1[M+1]
Step 2
4-Fluoro-7-hydroxy-2,3-dihydro-1H-inden-1-one 39c
[0518] Compound 39b (10 g, 49.4 mmol) and aluminum trichloride (20 g, 150 mmol, Sinopharm Chemical Reagent Co., Ltd.) were mixed well, and the reaction mixture was heated to 100° C., and stirred for 15 minutes. The reaction mixture was heated to 180° C. and reacted for 3 hours. The reaction mixture was cooled to room temperature, and slowly added to ice water. Ethyl acetate was added, and the resulting solution was stirred for 2 hours. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 39c (3 g, yield: 37%).
[0519] MS m/z (ESI): 167.3 [M+1]
Step 3
7-Fluoro-2,3-dihydro-1H-inden-4-ol 39d
[0520] Compound 39c (8 g, 48.2 mmol) was dissolved in trifluoroacetic acid (80 mL), followed by the addition of triethylsilane (14 g, 120.4 mmol). The reaction solution was stirred at 80° C. overnight. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 39d (6 g, yield: 82%).
[0521] MS m/z (ESI): 151.3 [M−1]
Step 4
5-Chloro-2-((7-fluoro-2,3-dihydro-1H-inden-4-yl)oxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 39
[0522] Compound 11b (1.02 g, 3.03 mmol), compound 39d (0.54 g, 3.03 mmol) and cesium carbonate (0.99 g, 3.13 mmol) were added to N-methylpyrrolidone (10 mL). The reaction solution was reacted at 60° C. overnight. The reaction solution was cooled, followed by the addition of ethyl acetate (150 mL), and washed with water (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative high performance liquid chromatography (Waters 2767-SQ Detecor2, eluent system: ammonium bicarbonate, water, acetonitrile) to obtain the title compound 39 (400 mg, yield: 29%).
[0523] MS m/z (ESI): 468.0 [M+1]
[0524] 1H NMR (400 MHz, DMSO-d.sub.6) δ 12.86 (s, 1H), 11.03 (s, 1H), 8.08 (s, 1H), 7.874-7.868 (d, 1H), 7.27 (m, 1H), 7.15 (m, 1H), 7.04-7.00 (m, 1H), 6.92-6.89 (m, 1H), 2.92-2.88 (m, 2H), 2.77-2.73 (m, 2H), 2.05-1.99 (m, 2H).
Example 40
5-Chloro-2-(2-(cyclopentyloxy)-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 40
[0525] ##STR00142##
[0526] In accordance with the synthetic route in Example 35, the starting compound deuterated methyl iodide in Step 1 was replaced with compound bromocyclopentane, accordingly, the title compound 40 (30 mg) was prepared.
[0527] MS m/z (ESI): 511.9[M+1].
[0528] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 312.87 (s, 1H), 10.99 (s, 1H), 8.05 (s, 1H), 7.95 (s, 1H), 7.30-7.35 (m, 1H), 7.29 (s, 1H), 7.11 (dd, 1H), 6.98 (s, 1H), 6.77-6.86 (m, 1H), 4.80-4.90 (m, 1H), 1.72-1.85 (m, 2H), 1.35-1.55 (m, 4H), 1.15-1.35 (m, 2H).
Example 41
5-Chloro-2-(2-cyclobutoxy-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 41
[0529] ##STR00143##
[0530] In accordance with the synthetic route in Example 35, the starting compound deuterated methyl iodide in Step 1 was replaced with compound bromocyclobutane, accordingly, the title compound 41 (110 mg) was prepared.
[0531] MS m/z (ESI): 498.0 [M+1]
[0532] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 12.87 (s, 1H), 11.02 (s, 1H), 8.06 (s, 1H), 7.94 (s, 1H), 7.29-7.23 (m, 2H), 7.09 (s, 1H), 6.95-6.92 (m, 1H), 6.92-6.83 (m, 1H), 4.73-4.70 (m, 1H), 2.37-2.33 (m, 2H), 1.79-1.72 (m, 2H), 1.68-1.52 (m, 2H).
Example 42
5-Chloro-2-(4-fluoro-2-isopropylphenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 42
[0533] ##STR00144##
[0534] In accordance with the synthetic route in Example 35, the starting compound deuterated methyl iodide in Step 1 was replaced with compound iodoisopropane, accordingly, the title compound 42 (100 mg) was prepared.
[0535] MS m/z (ESI): 485.9 [M+1]
[0536] 1H NMR (400 MHz, DMSO-d.sub.6): δ 12.87 (s, 1H), 11.00 (s, 1H), 8.05 (s, 1H), 7.95 (s, 1H), 7.32-7.16 (m, 3H), 7.01 (s, 1H), 6.86-6.82 (m, 1H), 4.67-4.62 (m, 1H), 1.08 (s, 3H), 1.06 (s, 3H).
Example 43
2-(2-Bromo-4-fluorophenoxy)-5-chloro-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 43
[0537] ##STR00145##
[0538] In accordance with the synthetic route in Example 11, the starting compound 2-methoxy-4-fluorophenol in Step 3 was replaced with 2-bromo-4-fluorophenol, accordingly, the title compound 43 (100 mg) was prepared.
[0539] MS m/z (ESI): 505.7 [M+1]
[0540] 1H NMR (400 MHz, DMSO-d.sub.6) 312.86 (s, 1H), 11.08 (s, 1H), 8.14 (s, 1H), 7.90 (d, 1H), 7.75 (dd, 1H), 7.36-7.26 (m, 3H), 7.17 (s, 1H).
Example 44
5-Chloro-2-(4-fluoro-2-(methyl-d.SUB.3.)phenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 44
[0541] ##STR00146##
Step 1
4-Fluoro-1-methoxy-2-(methyl-d.SUB.3.)benzene 44b
[0542] Compound 1-fluoro-4-methoxybenzene (5 g, 39.6 mmol, Accela ChemBio (Shanghai) Inc.) was dissolved in tetrahydrofuran (50 mL). The resulting solution was cooled to −10° C., and n-butyllithium (2.5 M, 43.7 mmol, 17.5 mL) was added dropwise. The reaction solution was naturally warmed up to room temperature and reacted for 1 hour. Iodomethane-d3 (5.8 g, 40 mmol, Accela ChemBio (Shanghai) Inc.) was slowly added dropwise under an ice bath, and the reaction solution was reacted at room temperature for 2 hours. Water (50 mL) was slowly added, and the reaction solution was extracted with ethyl acetate (20 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 44b (1.5 g, yield: 26%).
[0543] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.86-6.85 (m, 2H), 6.73-6.69 (m, 1H), 3.78 (s, 3H).
Step 2
4-Fluoro-2-(methyl-d.SUB.3.)phenol 44c
[0544] Compound 44a (1.5 g, 10.5 mmol) was added to dichloromethane (30 mL). A solution of boron tribromide in dichloromethane (1 M, 21.2 mmol, 21.2 mL) was added dropwise at room temperature, and the reaction solution was reacted at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 44c (900 mg, yield: 67%).
Step 3
5-Chloro-2-(4-fluoro-2-(methyl-d.SUB.3.)phenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 44
[0545] Compound 11b (100 mg, 0.3 mmol), compound 44c (39 mg, 0.3 mmol) and cesium carbonate (100 mg, 0.33 mmol) were added to N-methylpyrrolidone (2 mL). The reaction solution was reacted at 60° C. overnight. The reaction solution was cooled, followed by the addition of ethyl acetate (150 mL), and washed with water (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative high performance liquid chromatography (Waters 2767-SQ Detecor2, eluent system: ammonium bicarbonate, water, acetonitrile) to obtain the title compound 44 (21 mg, yield: 16%).
[0546] MS m/z (ESI): 445.0[M+1]
[0547] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.09 (s, 1H), 7.89-7.88 (d, 1H), 7.23-7.21 (d, 1H), 7.18-7.18 (dd, 1H), 7.13 (m, 1H), 7.10-7.08 (m, 2H).
Example 45
5-Chloro-2-(2-chloro-4-fluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 45
[0548] ##STR00147##
[0549] In accordance with the synthetic route in Example 11, the starting compound 2-methoxy-4-fluorophenol in Step 3 was replaced with 2-chloro-4-fluorophenol, accordingly, the title compound 45 (40 mg) was prepared.
[0550] MS m/z (ESI): 461.8 [M+1]
[0551] 1H NMR (400 MHz, DMSO-d.sub.6) δ 12.87 (s, 1H), 11.08 (s, 1H), 8.14 (s, 1H), 7.89 (d, 1H), 7.64 (dd, 1H), 7.33-7.26 (m, 3H), 7.17 (s, 1H).
Example 46
5-Chloro-2-(2,4-difluorophenoxy)-N-(6-oxo-1,6-dihydropyridazin-4-yl)-4-(trifluoromethyl)benzamide 46
[0552] ##STR00148##
[0553] In accordance with the synthetic route in Example 11, the starting compound 2-methoxy-4-fluorophenol in Step 3 was replaced with 2,4-difluorophenol, accordingly, the title compound 46 (70 mg) was prepared.
[0554] MS m/z (ESI): 445.8 [M+1]
[0555] 1H NMR (400 MHz, DMSO-d.sub.6) 312.87 (s, 1H), 11.10 (s, 1H), 8.12 (s, 1H), 7.89 (d, 1H), 7.53-7.47 (m, 1H), 7.38-7.32 (m, 2H), 7.18-7.12 (m, 2H).
Biological Assay
[0556] The present disclosure will be further described with reference to the following test examples, but the examples should not be considered as limiting the scope of the present disclosure.
Test Example 1. Determination of the Inhibitory Activity of the Compounds of the Present Disclosure on Nav1.8
[0557] The purpose of the experiment is to investigate the effect of the compounds on Nav1.8 ion channel in an in vitro experiment, wherein the Nav1.8 ion channel is stably expressed on HEK293 cells. After the Nav1.8 current becomes stable, the Nav1.8 currents before and after the administration of the compound are compared so as to obtain the effect of the compound on the Nav1.8 ion channel.
[0558] 1. Experimental Materials and Instruments
[0559] 1) Patch clamp amplifier: patch clamp PC-505B (WARNER instruments)/MultiClamp 700A (Axon instrument)
[0560] 2) Digital-to-analog converter: Digidata 1440A (Axon CNS)/Digidata 1550A (Axon instruments)
[0561] 3) Micro-manipulator: MP-225 (SUTTER instrument)
[0562] 4) Inverted microscope: TL4 (Olympus)
[0563] 5) Glass microelectrode puller: PC-10 (NARISHIGE)
[0564] 6) Microelectrode glass capillary: B12024F (Wuhan Weitan Scientific Instrument Co., Ltd.)
[0565] 7) Dimethyl sulfoxide (DMSO) D2650 (Sigma-Aldrich)
[0566] 8) TTX AF3014 (Affix Scientific)
[0567] 2. Experimental Procedures
[0568] 2.1 Formulation of the Compounds
[0569] Except for NaOH and KOH used for acid titration and base titration, all the compounds used for formulating the extracellular fluid and intracellular fluid were purchased from Sigma (St. Louis, Mo.). Extracellular fluid (mM): NaCl, 137; KCl, 4; CaCl.sub.2), 1.8; MgCl.sub.2, 1; HEPES, 10; glucose, 10; pH 7.4 (NaOH titration). Intracellular fluid (mM): aspartic acid, 140; MgCl.sub.2, 2; EGTA, 11; HEPES, 10; pH 7.2 (CsOH titration). All solutions of test compound and control compound contained 1 μM TTX.
[0570] The test compound was dissolved in dimethyl sulfoxide (DMSO) at a stock concentration of 9 mM. The stock solution of the test compound was dissolved in the extracellular fluid on the day of the test and formulated into the required concentration.
[0571] 2.2 Test Process of the Manual Patch Clamp
[0572] 1) The compound was formulated into solutions with specified concentrations, the solutions were added to the pipelines respectively in order from low to high concentration, and the pipelines were marked.
[0573] 2) The cell was transferred to the perfusion tank. A positive pressure was applied to the electrode. The tip of the electrode touched the cell. The three-way valve of the air extracting device was adjusted to a three-way state. A negative pressure was applied to the electrode, so that a high-resistance seal was formed between the electrode and the cell.
[0574] The negative pressure was applied continuously, thereby causing the cell membrane to rupture and forming a current path.
[0575] 3) After the current for rupturing the cell membrane became stable, perfusion of different concentrations was carried out in sequence. Once the current was stable for at least one minute, perfusion of the next concentration was carried out. The duration of the perfusion of each concentration did not exceed five minutes.
[0576] 4) The perfusion tank was cleaned. The perfusion tank was rinsed with the drug solutions in order from high to low concentration, and the rinse duration for each concentration of drug solution was 20 seconds. The perfusion tank was finally rinsed with the extracellular fluid for 1 minute.
[0577] 2.3 Test Voltage Equation (Resting) and Results
[0578] The cell was clamped at −80 mV. The cell was depolarized to 10 mV with a square wave lasting 10 milliseconds to obtain the Nav1.8 current. This procedure was repeated every 5 seconds. The maximum current caused by the square wave was measured. After the current became stable, the test compound was perfused. After the response became stable, the blocking intensity was calculated.
[0579] 3. Data Analysis
[0580] The data was stored in the computer system for analysis. Data collection and analysis were carried out by pCLAMP 10 (Molecular Devices, Union City, Calif.), and the analysis results were reviewed by the administrator. Stable current means that the current changes within a limited range over time. The magnitude of stable current was used to calculate the effect of the compound at the concentration.
[0581] The inhibitory activity of the compounds of the present disclosure on Nav1.8 was determined by the above test, and the resulting IC.sub.50 values are shown in Table 1.
TABLE-US-00003 TABLE 1 IC.sub.50 of the compounds of the present disclosure on inhibiting the Nav1.8 channel activity Example No. IC.sub.50 (nM) 1 1.6 2 1.3 3 3.3 4 14.1 5 23.7 6 24.4 7 30.8 8 45.8 9 83.4 11 1.3 12 0.26 13 2.3 14 2.5 15 4.7 16 17.9 17 18.0 18 22.2 19 31.7 20 58.5 21 59.4 22 87.8 23 92.3 27 5.7 28 5.2 29 90.4 30 16.2 31 0.94 32 0.37 33 0.86 34 11.5 35 0.54 36 1.54 37 2.38 39 0.24 40 0.32 41 1.86 42 2.98 43 3.36 44 4.04 45 5.08 46 8.95
[0582] Conclusion: The compounds of the present disclosure have a significant inhibitory effect on the Nav1.8 channel activity.
Pharmacokinetics Evaluation
Test Example 2. Pharmacokinetics Assay of the Compounds of the Present Disclosure
[0583] 1. Abstract
[0584] Rats were used as test animals. The drug concentration in plasma at different time points was determined by LC/MS/MS method after intragastrical administration of the compounds of Example 2, Example 11, Example 12, Example 15, Example 31 and Example 33 to rats. The pharmacokinetic behavior of the compounds of the present disclosure was studied and evaluated in rats.
[0585] 2. Test Protocol
[0586] 2.1 Test Compounds
[0587] Compounds of Example 2, Example 11, Example 12, Example 15, Example 31 and Example 33.
[0588] 2.2 Test Animals
[0589] Twenty-four healthy adult SD rats (half male and half female, equally divided into six groups, four rats per group) were purchased from Shanghai Jiesijie Laboratory Animal Co., LTD.
[0590] 2.3 Preparation of the Test Compound
[0591] A certain amount of the test compound was weighed, to which 5% of DMSO, 5% of tween 80 and 90% of normal saline were added to prepare a 0.2 mg/mL colorless, clear and transparent solution.
[0592] 2.4 Administration
[0593] After an overnight fast, SD rats were intragastrically administered the test compound at an administration dose of 2.0 mg/kg and an administration volume of 10.0 mL/kg.
[0594] 3. Process
[0595] The rats were intragastrically administered the compounds of Example 2, Example 11, Example 12, Example 15, Example 31 and Example 33. 0.2 ml of blood was taken from the orbit before the administration and at 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 11.0 and 24.0 hours after the administration. The samples were stored in heparinized tubes, and centrifuged for 10 minutes at 10000 rpm and 4° C. to separate the blood plasma. The plasma samples were stored at −20° C. The rats were fed 2 hours after the administration.
[0596] The content of the test compound in the plasma of rats after intragastrical administration of the test compound at different concentrations was determined: 25 μL of rat plasma at each time point after the administration was taken, to which 30 μL of the internal standard solution and 175 μL of acetonitrile were added. The resulting solution was vortex-mixed for 5 minutes, and centrifuged for 10 minutes (3700 rpm). 0.5 μL of the supernatant was taken from the plasma samples for LC/MS/MS analysis.
[0597] 4. Results of Pharmacokinetic Parameters
[0598] Pharmacokinetic parameters of the compounds of the present disclosure are shown below.
TABLE-US-00004 Pharmacokinetics assay (2 mg/kg) Plasma Area under Apparent Bio- concentration curve Residence Clearance distribution avail- Cmax AUC Half-life time CL/F volume ability No. (ng/mL) (ng/mL * h) T½ (h) MRT(h) (ml/min/kg) Vz/F (ml/kg) F (%) 2 399 ± 153 5963 ± 4279 47 ± 54.6 68.2 ± 78.3 6.24 ± 6.29 4711 ± 1293 149 11 218 ± 103 1234 ± 621 2.3 ± 1.14 4..36 ± 1.46 32.3 ± 15.2 5332 ± 720 98 12 435 ± 64 5981 ± 1126 13.2 ± 7.8 19.8 ± 10.4 4.3 ± 1.65 4174 ± 587 111 15 295 ± 44 5689 ± 552 56.7 ± 6.6 82.4 ± 10.2 1.46 ± 0.11 7154 ± 682 94 31 360 ± 65 5619 ± 1233 33.3 ± 34.8 48.3 ± 50.3 3.77 ± 2.70 5436 ± 705 66 33 382 ± 88 5554 ± 3021 18.6 ± 17.5 27.7 ± 24.2 6.4 ± 5.57 3990 ± 788 99 35 392 ± 43 4328 ± 910 7.5 ± 1.92 10.6 ± 2.3 7.93 ± 1.47 5008 ± 875 107
[0599] Conclusion: The compound of the present disclosure is well absorbed, and has a significant pharmacokinetic advantage.