Patent Application
20240132445
The present invention relates to the field of medicinal chemistry, in particular to a phenol derivative, a stereoisomer and pharmaceutically acceptable salt thereof, and the use of the compound of the present invention in the preparation of a medicament for treating central nervous system-related diseases.
A GABAA receptor is a receptor of the chief inhibitory neurotransmitter in the central nervous system. The GABAA receptor is composed of a pentamer of transmembrane polypeptide subunits, and 19 different subunits assemble into various GABAA receptor subtypes. The GABAA receptor is involved in pathogenesis, diagnosis and treatment of various diseases such as anesthesia, depression, anxiety, epilepsy, memory disorder and drug dependence. Accordingly, the GABAA receptor has become a pharmacologically and clinically important drug target. Propofol and its derivatives represent a class of important GABAA-targeting compounds.
Propofol can activate many GABAA receptor subtypes, and is widely used for inducing and maintaining general anesthesia. Propofol shows remarkable pharmacokinetic and pharmacodynamic characteristics in that it rapidly takes effect, acts for a short period, and is quickly reversible. Upon intravenous administration, propofol in the blood rapidly enters hyperperfused areas such as heart, lung and liver, and its high liposolubility allows propofol to easily travel across the blood-brain barrier into the brain for general anesthesia.
With the in-depth clinical application of propofol, many of its limitations and disadvantages have been reported one after another. It has been reported that approximately 70% of patients on propofol injections feel certain pain or discomfort. It has been demonstrated that propofol can lower the systolic pressure, the diastolic pressure and the mean arterial pressure, and thus may clinically cause hypotension. Furthermore, respiratory depression is also an unneglectable risk upon use of propofol. These adverse effects have considerably impeded application of propofol in certain clinical cases, such as cardiovascular diseases, brain injury and chronic hypotension.
Fospropofol disodium is a water-soluble prodrug of propofol, and can be rapidly hydrolysed by alkaline phosphatase to release propofol, phosphate and formaldehyde. Although fospropofol disodium relieves pain at sites of intravenous propofol injection, it still poses risks of respiratory depression and adverse hemodynamic effects because it takes effect in the form of the active compound propofol. In addition, fospropofol disodium may also cause abnormal sensation and itching.
With regard to the limitations and disadvantages of propofol and fospropofol described above, there is a need for developing a novel GABAA receptor agonist with better pharmacokinetic and pharmacodynamic characteristics and fewer side effects.
The patent US 20050032753 A1 describes a phenol derivative useful for anesthesia and sedation, wherein R1 and R2 are independently selected from C1-8 alkyl and C1-8 cycloalkyl; L is selected from a covalent bond or C1-12 hydrocarbylene; and R3 is selected from —C(═O)ORa, wherein Ra is selected from C1-12 hydrocarbylene.
The patent further discloses the following formula, wherein R4 is C1-C5 alkyl, C2-C5 alkenyl or C2-C5 alkynyl; R5 is C1-C6 alkyl or C3-C8 cycloalkyl; R6 is methyl; or R5 and R6 together with the carbon atoms to which they are attached form C3-8 cycloalkyl; and Ra is C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl or C3-C8 cycloalkyl. However, none of the compounds disclosed in the patent is included in the general structural formula of the present invention, and the patent does not disclose pharmacodynamic data for any specific compound. Those skilled in the art would not have been able to know from the patent whether different substituents result in differences in efficacy, and also would not have been motivated by the patent to make further improvement in a specific direction, to obtain specific compound structures with better efficacy. The compound of the patent greatly differs in structure from the compound of the present invention, and the specific descriptions in the patent are not considered as a part of the present invention.
The patent CN 104507899 A discloses a phenol derivative, a preparation method therefor, and the use thereof in the central nervous field. The patent discloses the following general formula. The compound of the patent greatly differs in structure from the compound of the present invention, and the specific descriptions in the patent are not considered as a part of the present invention.
The patent CN 104507898 A discloses a phenol derivative, a preparation method therefor, and the use thereof in the central nervous field. The patent discloses the following general formula. The compound of the patent greatly differs in structure from the compound of the present invention, and the specific descriptions in the patent are not considered as a part of the present invention.
An objective of the present invention is to provide a GABAA receptor agonist phenol derivative that has a novel structure and better efficacy, can effectively reduce side effects, and is safer for clinical use, a stereoisomer thereof, and the use thereof in the central nervous field, thereby providing more and better choices for medicaments for inducing and/or maintaining anesthesia in animal or human bodies, facilitating sedation and hypnosis, and treating and/or preventing anxiety, nausea, vomiting, migraine, convulsion, epilepsy, neurodegenerative diseases, and central nervous system-related diseases.
In one aspect, the present invention provides a compound as represented by general formula (I), or a stereoisomer or pharmaceutically acceptable salt thereof:
—(CH2) mCOOR12,
or C1-10 alkyl, wherein the alkyl is optionally further substituted with one or more R;
In some preferred embodiments provided by the present invention, the compound is selected from a compound as represented by general formula (II):
In another preferred embodiment provided by the present invention, the compound is selected from a compound as represented by general formula (III):
Preferably,
Preferably,
Preferably,
R is selected from F, Cl, Br or I.
Preferably,
In another preferred embodiment of the present invention, the following compounds are included:
In another aspect, the present invention provides a pharmaceutical composition, comprising the compound or the stereoisomer or pharmaceutically acceptable salt thereof described above in the present invention, and one or more pharmaceutically acceptable carriers.
The pharmaceutical composition involved in the present invention is in any one of the pharmaceutically acceptable dosage forms, such as tablets, capsules, dispersible tablets, granules, injections, lipid emulsions, aerosols, inhalation powders, sprays, oral solutions, and oral suspensions.
The present invention also provides the use of the compound or the stereoisomer or pharmaceutically acceptable salt thereof and the pharmaceutical composition thereof described in the present invention in the preparation of a medicament for inducing and/or maintaining anesthesia in animal or human bodies, facilitating sedation and hypnosis in animal or human bodies, and treating and/or preventing anxiety, depression, insomnia, nausea, vomiting, migraine, schizophrenia, convulsion and epilepsy.
Unless stated to the contrary, the terms used in the description and claims have the following meanings.
“Alkyl” refers to a straight or branched saturated aliphatic hydrocarbon group containing 1 to 20 carbon atoms, preferably alkyl containing 1 to 8 carbon atoms, more preferably alkyl containing 1 to 6 carbon atoms, further preferably alkyl containing 1 to 4 carbon atoms. Examples include methyl, ethyl, n-propyl, isopropyl, etc.
“Alkoxy” refers to —O-alkyl. Examples include methoxy, ethoxy, n-propoxy, isopropoxy, etc.
“Cycloalkoxy” refers to a group formed by bonding the above-mentioned cycloalkyl to an oxygen atom. As 3- to 6-membered cycloalkoxy, examples include cyclopropyloxy, cyclohexyloxy, etc.
“Cycloalkyl” refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, and a cycloalkyl ring comprises 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 8 carbon atoms, and most preferably 3 to 6 (for example, 3, 4, 5 or 6) carbon atoms. Non-limiting examples of monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc. Examples of polycyclic cycloalkyl include spiro ring, fused ring and bridged ring cycloalkyl.
“Heterocyclyl” refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, which comprises 3 to 20 ring atoms, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O)m (wherein m is an integer of 0 to 4), but excluding ring moieties of —O—O—, —O—S— or —S—S—, and the remaining ring atoms are carbon. The cyclic hydrocarbon substituent preferably comprises 3 to 12 ring atoms, of which 1-4 are heteroatoms. The cyclic hydrocarbon substituent preferably comprises 3 to 8 ring atoms, of which 1-3 are heteroatoms. The cyclic hydrocarbon substituent preferably comprises 3 to 6 ring atoms, of which 1-3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyl include azetidinyl, pyrrolidinyl, imidazolidinyl, tetrahydrofuryl, tetrahydropyranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuryl, dihydropyrazolyl, dihydropyrrolyl, piperidyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc., preferably tetrahydropyranyl, piperazinyl, and pyrrolidinyl. Examples of polycyclic heterocyclyl include spiro ring, fused ring and bridged ring heterocyclyl.
“ Stereoisomer” refers to an isomer produced as a result of different spatial arrangements of atoms in molecules, including cis-trans isomers, enantiomers and conformational isomers.
“Optional” or “optionally” or “alternative” means that the events or conditions subsequently described may but not necessarily occur, and the description includes the case where the events or conditions occur and do not occur. For example, “heterocyclyl alternatively substituted with alkyl” means that the alkyl may but not necessarily exist, and the description includes the case where the heterocyclyl is substituted with alkyl and the case where the heterocyclyl is not substituted with alkyl.
The implementation process and beneficial effects of the present invention are described in detail below by way of specific examples, which are intended to help those skilled in the art better understand the essence and characteristics of the present invention, and are not intended to limit the scope of implementation of the present invention.
Compound 1-1 (10 g, 73.43 mmol, 1.0 eq) and DMAP (897 mg, 9.34 mmol) were added to dichloromethane (200 mL). Acetic anhydride (9.74 g, 95.45 mmol, 1.3 eq) was added dropwise at 15° C. After the dropwise addition was completed, the mixture was slowly warmed to room temperature (15° C.) and reacted for 16 h. The reaction liquid was neutralized with 1 N hydrochloric acid solution to pH=6-7, and then extracted with dichloromethane (150 mL×3). The organic phases were combined and subjected to rotary evaporation to remove the solvent. Column separation was conducted (eluent:VEthyl acetate:VPetroleum ether=1:10) to obtain compound 1-2 as a yellow transparent oil (13.16 g, yield: 99.1%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.34-7.31 (m, 1H), 7.24-7.18 (m, 2H), 7.01-6.98 (m, 1H), 3.05-3.01 (m, 1H), 2.33 (s, 3H), 1.23 (d, J=8.0 Hz, 6H).
Compound 1-2 (13.09 g, 73.45 mmol, 1.0 eq) was mixed with aluminium trichloride (10.58 g, 79.32 mmol, 1.1 eq), and then the reaction mixture was heated to 140° C. and reacted for 5 hours. After TLC detection showed that the reaction was completed, the reaction liquid was poured into saturated NH4Cl (200 mL), and filtration and liquid separation were conducted. The organic phase was subjected to rotary evaporation to remove the solvent, and column separation (eluent:VPetroleum ether:VEthyl acetate=50:1 to 20:1) was conducted to obtain compound 1-3 as a yellow transparent oil (1.61 g, yield: 12.3%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ12.70 (s, 1H), 7.61-7.59 (m, 1H), 7.43-7.41 (m, 1H), 6.89-6.85 (m, 1H), 3.40-3.35 (m, 1H), 2.64 (s, 3H), 1.26-1.23 (m, 6H).
Compound 1-3 (800 mg, 4.49 mmol, 1.0 eq) was added to dry methanol (15 mL), and then NaBH4 (255 mg, 6.73 mmol, 1.5 eq) was added. The mixture was reacted at room temperature for 0.5 hours. The reaction liquid was poured into a saturated ammonium chloride aqueous solution (10 mL), and extracted with DCM (50 mL×4). The organic phases were combined and subjected to rotary evaporation to remove the solvent, so as to obtain compound 1-4 as a yellow oil (809 mg, yield: 100%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ8.13 (s, 1H), 7.15-7.13 (m, 1H), 6.85-6.81 (m, 2H), 5.10-5.04 (m, 1H), 3.41-3.32 (m, 1H), 2.40 (s, 1H), 1.62 (d, J=8.0 Hz, 3H), 1.28-1.20 (m, 6H).
Compound 1-4 (0.75 g, 4.16 mmol) was added to methanethiol (2.40 g, 10% propylene glycol solution, 4.99 mmol, 1.2 eq) under nitrogen protection, and hydrochloric acid (230 mg, 6.24 mmol, 1.5 eq) was added dropwise to the reaction liquid. The reaction system was stirred and reacted at 25° C. for 16 hours. After TLC (VHexane:VEA=5:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with H2O and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated saline and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=15:1 to 12:1) to obtain compound 1-5 as a yellow oil (440 mg, yield: 50.3%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.38 (s, 1H), 7.17-7.15 (m, 1H), 6.92-6.90 (m, 1H), 6.85-6.82 (m, 1H), 4.07-4.02 (m, 1H), 3.40-3.32 (m, 1H), 1.94 (d, J=4.0 Hz, 3H), 1.65 (d, J=8.0 Hz, 3H), 1.26-1.21 (m, 6H).
Under nitrogen protection, compound 1-5 (140 mg, 0.666 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (240 mg, 1.332 mmol, 2.0 eq) was slowly added to the reaction, and the reaction was conducted at 0° C.-5° C. for 1.5 hours. After TLC (VHexane:VEA=2:1) detection showed that the raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (20 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=10:1) to obtain compound 1 as a yellow oil (60.0 mg, yield: 37.3%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.26-7.24 (m, 1H), 7.12-7.10 (m, 1H), 7.01-6.97 (m, 1H), 6.85 (s, 1H), 4.63-4.58 (m, 1H), 3.37-3.30 (m, 1H), 2.76 (s, 3H), 1.83 (d, J=8.0 Hz, 3H), 1.26-1.23 (m, 6H).
Under nitrogen protection, compound 1-5 (100 mg, 0.475 mmol) was dissolved in DCM (5 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (48.62 mg, 0.237 mmol, 0.5 e.q) was slowly added to the reaction, and the reaction was conducted at 0° C.-5° C. for 0.5 hours. After TLC (VHexane:VEA=1:1) detection showed that the reaction of the raw materials was almost completed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (20 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:1) to obtain compound 2 as a colourless oil (28.0 mg, yield: 26.0%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ9.67 (s, 1H), 7.20-7.18 (m, 1H), 6.86-6.82 (m, 1H), 6.80-6.78 (m, 1H), 3.76-3.70 (m, 1H), 3.45-3.38 (m, 1H), 2.45 (s, 3H), 1.89 (d, J=8.0 Hz, 3H), 1.24-1.21 (m, 6H).
Compound 3-1 (5.00 g, 73.43 mmol, 1.0 e.q) and NaOH (2.94 g, 73.42 mmol) were added to DMF (80 mL), and the mixture was stirred at 15° C. for 30 min. 1-chloro-2-butylene (a cis-trans mixture, 4.32 g, 47.74 mmol, 1.3 e.q) was then slowly added dropwise. After the dropwise addition was completed, the system was reacted at 15° C. for additional 16 hours. After TLC detection showed that the reaction was completed, the reaction liquid was slowly poured into ice water (200 mL), and extracted with n-hexane (200 mL×3). The organic phases were combined, and subjected to rotary evaporation to remove the solvent, so as to obtain compound 3-2 as a yellow transparent oil (6.99 g, yield: 100%).
Compound 3-2 (6.99 g, 36.72 mmol, 1.0 eq) was added to a 50-mL single-necked flask. Under nitrogen protection, condensation was conducted with air reflux. The reaction mixture was heated to 210° C.-215° C. and reacted for 3.5 hours. After TLC detection showed that the reaction was completed, the reaction liquid was diluted with ethyl acetate, and subjected to sample stirring and column separation (eluent:VPetroleum ether:VEthyl acetate=50:1/20:1) to obtain compound 3-3 as a yellow transparent oil (1.60 g, yield: 22.9%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ12.70 (s, 1H), 7.63-7.61 (m, 1H), 7.39-7.36 (m, 1H), 6.88-6.85 (m, 1H), 6.05-6.00 (m, 1H), 5.11-5.05 (m, 2H), 3.98-3.97 (m, 1H), 2.64 (s, 3H), 1.34 (d, J=4.0 Hz, 3H).
Under nitrogen protection, a 100-mL three-necked flask was subjected to replacement 3-5 times, and DCM (10 mL) was added. The reaction system was cooled to −5═ C.-0° C., and diethyl zinc (9.46 mL, 2.0 M, 18.92 mmol) was slowly added dropwise to the reaction liquid, with the addition completed in about 10 minutes. Then trifluoroacetic acid (2.16 mg, 18.92 mmol) was added at 5° C.-0° C. (ice ethanol bath). After about 5 minutes, diiodomethane (6.76 g, 25.23 mmol) was dissolved in DCM (5 mL), and the mixture was added to the reaction liquid using a syringe. The system temperature was kept at 5° C.-0° C. After 60 minutes, compound 3-3 (1.2 g, 6.31 mmol) was dissolved in DCM (5 mL), and the mixture was slowly added dropwise to the reaction. After the dropwise addition was completed, the ice bath was removed. The reaction was warmed to 25° C. and stirred for additional 48 hours. After 1HNMR and TLC (VHexane:VEA=10:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (20 mL), and washed with a saturated ammonium chloride solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=60:1 to 15:1) to obtain compound 3-4 as a yellow oil (1.1 g, yield: 85.3%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ12.63 (s, 1H), 7.63-7.60 (m, 1H), 7.56-7.54 (m, 1H), 6.91-6.87 (m, 1H), 2.64 (s, 3H), 2.52-2.48 (m, 1H), 1.34-1.27 (m, 3H), 1.03-0.99 (m, 1H), 0.56-0.50 (m, 1H), 0.38-0.30 (m, 1H), 0.24-0.20 (m, 1H), 0.17-0.15 (m, 1H).
Compound 3-4 (1.1 g, 5.39 mmol, 1.0 eq) was added to dry methanol (15 mL), and then NaBH4 (265 mg, 7.00 mmol, 1.5 eq) was added in portions. The mixture was reacted at room temperature for 0.5 hours. The reaction liquid was poured into a saturated ammonium chloride aqueous solution (10 mL), and extracted with DCM (50 mL×2). The organic phases were combined and concentrated to remove the solvent, so as to obtain compound 3-5 as a yellow transparent oil (1.10 g, yield: 99.1%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ8.08 (s, 1H), 7.26-7.24 (m, 1H), 7.02-6.98 (m, 1H), 6.86-6.83 (m, 1H), 5.28-5.06 (m, 1H), 2.53-2.46 (m, 1H), 1.62-1.60 (m, 3H), 1.31-1.26 (m, 3H), 1.03-0.99 (m, 1H), 0.56-0.54 (m, 1H), 0.39-0.37 (m, 1H), 0.22-0.20 (m, 1H), 0.17-0.15 (m, 1H).
Compound 3-5 (1.3 g, 6.30 mmol) was added to acetonitrile (15 mL), and methanethiol (3.94 g, 10% propylene glycol solution, 8.19 mmol, 1.2 e.q.) was added under nitrogen protection. Then hydrochloric acid (344 mg, 9.45 mmol, 1.5 eq) was added dropwise to the reaction liquid. The reaction system was stirred and reacted at 25° C for 4 hours. After TLC (VHexane:VEA=5:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with H2O and extracted with ethyl acetate (50 mL×3) . The organic phases were combined, washed with saturated saline and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=50:1 to 20:1) to obtain compound 3 as a yellow oil (500 mg, yield: 33.6%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.35-7.34 (m, 1H), 7.27-7.25 (m, 1H), 6.93-6.91 (m, 1H), 6.87-6.83 (m, 1H), 4.09-4.02 (m, 1H), 2.57-2.52 (m, 1H), 1.94(d, J=8.0 Hz, 3H), 1.65 (d, J=8.0 Hz, 3H), 1.30-1.24 (m, 3H), 1.04-0.95 (m, 1H), 0.53-0.50 (m, 1H), 0.35-0.30 (m, 1H), 0.20-0.15 (m, 2H).
Under nitrogen protection, compound 3 (100 mg, 0.423 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (165 mg, 0.846 mmol, 2.0 eq) was slowly added to the reaction, and the reaction was conducted at 15° C. for 0.5 hours. After TLC (VHexane:VEA=1:1) detection showed that the raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:1) to obtain compound 4 as a yellow oil (40.0 mg, yield: 35.4%).
Characterization data: 1NMR (400 MHz, CDCl3) δ7.35-7.34 (m, 1H), 7.14-7.12 (m, 1H), 7.02-6.98 (m, 1H), 6.84 (s, 1H), 4.65-4.59 (m, 1H), 2.76 (s, 3H), 2.59-2.54 (m, 1H), 1.83 (d, J=8.0 Hz, 3H), 1.30-1.25 (m, 3H), 1.06-0.99 (m, 1H), 0.60-0.54 (m, 1H), 0.46-0.40 (m, 1H), 0.26-0.13 (m, 2H).
Under nitrogen protection, compound 3 (100 mg, 0.423 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (77.30 mg, 0.381 mmol, 0.9 eq) was slowly added to the reaction, and the reaction was conducted at 0° C.-5° C. for 0.5 hours. After TLC (VHexane:VEA=1:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:1) to obtain compound 5 as a yellow oil (40.0 mg, yield: 37.7%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ9.61 (s, 1H), 7.32-7.30 (m, 1H), 6.88-6.79 (m, 2H), 3.77-3.71 (m, 1H), 2.59-2.56 (m, 1H), 2.45 (s, 3H), 1.88 (d, J=8.0 Hz, 3H), 1.30-1.28 (m, 3H), 1.04-1.01 (m, 1H), 0.53-0.50 (m, 1H), 0.37-0.32 (m, 1H), 0.21-0.16 (m, 2H).
Compound 3-5 (1.0 g, 4.85 mmol) was added to acetonitrile (10 mL), and isopropylthiol (443.0 mg, 5.82 mmol, 1.2 e.q.) was added under nitrogen protection. Then hydrochloric acid (230 mg, 6.30 mmol, 1.3 e.q.) was added dropwise to the reaction liquid. The reaction system was stirred and reacted at 15° C. for 16 hours. After TLC (VHexane:VEA=5:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with H2O and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated saline and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=50:1 to 20:1) to obtain compound 6-1 as a yellow oil (600 mg, yield: 46.8%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.70 (d, J=8.0 Hz, 1H), 7.26-7.24 (m, 1H), 6.92-6.90 (m, 1H), 6.86-6.82 (m, 1H), 4.20-4.15 (m, 1H), 2.70-2.63 (m, 1H), 2.58-2.51 (m, 1H), 1.63 (d, J=8.0 Hz, 3H), 1.30-1.24 (m, 3H), 1.17-1.14 (m, 6H), 1.04-0.99 (m, 1H), 0.56-0.52 (m, 1H), 0.39-0.34 (m, 1H), 0.22-0.15 (m, 2H).
Under nitrogen protection, compound 6-1 (100 mg, 0.378 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (154 mg, 85%, 0.756 mmol, 2.0 eq) was slowly added to the reaction, and the reaction was conducted at 15° C. for 0.5 hours. After TLC (VHexane:VEA=1:1) detection showed that the raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:1) to obtain compound 6 as a yellow oil (50.0 mg, yield: 44.6%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.35-7.31 (m, 1H), 7.10-7.08 (m, 1H), 6.97-6.93 (m, 1H), 4.60-4.58 (m, 1H), 3.18-3.10 (m, 1H), 2.60-2.54 (m, 1H), 1.81(d, J=8.0 Hz, 3H), 1.31-1.27 (m, 6H), 1.26-1.17 (m, 3H), 1.04-0.98 (m, 1H), 0.56-0.52 (m, 1H), 0.37-0.34 (m, 1H), 0.22-0.15 (m, 2H).
Under nitrogen protection, compound 6-1 (100 mg, 0.378 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (69.1 mg, 0.341 mmol, 0.9 eq) was slowly added to the reaction, and the reaction was conducted at 0° C.-5° C. for 0.5 hours. After TLC (VHexane:VEA=1:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:1) to obtain compound 7 as a yellow oil (30.0 mg, yield: 28.3%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.35-7.31 (m, 1H), 7.10-7.08 (m, 1H), 6.97-6.93 (m, 1H), 4.60-4.58 (m, 1H), 2.68-2.63 (m, 1H), 2.45-2.38 (m, 1H), 1.81 (d, J=8.0 Hz, 3H), 1.31-1.27 (m, 6H), 1.26-1.17 (m, 3H), 1.04-0.98 (m, 1H), 0.56-0.54 (m, 1H), 0.39-0.32 (m, 1H), 0.22-0.17 (m, 2H).
Compound 3-5 (1.0 g, 4.85 mmol) was added to acetonitrile (10 mL), and cyclopropylthiol (431.2 mg, 5.82 mmol, 1.2 eq) was added under nitrogen protection. Then concentrated hydrochloric acid (230 mg, 6.30 mmol, 1.3 eq) was added dropwise to the reaction liquid. The reaction system was stirred and reacted at 15° C. for 12 hours. After TLC (VHexane:VEA=5:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with H2O and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated saline and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=50:1 to 20:1) to obtain compound 8-1 as a yellow oil (490 mg, yield: 38.5%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.35-7.34 (m, 1H), 7.27-7.25 (m, 1H), 6.93-6.91 (m, 1H), 6.87-6.83 (m, 1H), 4.09-4.02 (m, 1H), 2.57-2.52 (m, 1H), 2.05-2.00 (m, 1H), 1.94 (d, J=8.0 Hz, 3H), 1.65 (d, J=8.0 Hz, 3H), 1.04-0.95 (m, 2H), 0.74-0.58 (m, 2H), 0.59-0.50 (m, 1H), 0.39-0.30 (m, 2H), 0.24-0.12 (m, 2H).
Under nitrogen protection, compound 8-1 (100 mg, 0.381 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (155 mg, 0.762 mmol, 2.0 eq) was slowly added to the reaction, and the reaction was conducted at 15° C. for 0.5 hours. After TLC (VHexane:VEA=1:1) detection showed that the raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:1) to obtain compound 8 as a yellow solid (60.0 mg, yield: 53.4%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.35-7.34 (m, 1H), 7.14-7.12 (m, 1H), 7.02-6.98 (m, 1H), 6.84 (s, 1H), 4.65-4.59 (m, 1H), 2.59-2.54 (m, 1H), 2.45-2.35 (m, 1H), 1.83 (d, J=8.0 Hz, 3H), 1.30-1.24 (m, 3H), 1.04-0.95 (m, 1H), 0.83-0.80 (m, 1H), 0.65-0.60 (m, 2H), 0.53-0.50 (m, 1H), 0.45-0.40 (m, 1H), 0.35-0.30 (m, 1H), 0.20-0.15 (m, 2H).
Under nitrogen protection, compound 8-1 (100 mg, 0.381 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (69.6 mg, 0.343 mmol, 0.9 eq) was slowly added to the reaction, and the reaction was conducted at 0° C.-5° C. for 0.5 hours. After TLC (VHexane:VEA=1:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:1) to obtain compound 9 as a yellow transparent oil (38.0 mg, yield: 35.8%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ9.63 (s, 1H), 7.35-7.32 (m, 1H), 6.89-6.75 (m, 2H), 3.77-3.71 (m, 1H), 2.59-2.56 (m, 1H), 2.45-2.34 (m, 1H), 1.88 (d, J=8.0 Hz, 3H), 1.30-1.24 (m, 3H), 1.04-0.95 (m, 1H), 0.83-0.80 (m, 1H), 0.65-0.60 (m, 2H), 0.53-0.50 (m, 1H), 0.45-0.40 (m, 1H), 0.350.30 (m, 1H), 0.20-0.15 (m, 2H).
Compound 1-3 (2.00 g, 11.22 mmol, 1.0 eq) was added to a 100-mL single-necked flask, dry methanol (40 mL) and potassium carbonate (2.33 g, 16.83 mmol, 1.5 eq) were added, and then benzyl chloride (1.85 g, 14.59 mmol, 1.3 eq) was added. The reaction mixture was heated to 30° C. and reacted for 2 h under nitrogen protection. After TLC detection showed that the reaction was completed, the reaction liquid was diluted with ethyl acetate, and subjected to sample stirring and column separation (eluent:VPetroleum ether:VEthyl acetate=100:1 to 50:1) to obtain compound 10-1 as a yellow oil (2.5 g, yield: 83.3%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.66-7.64 (m, 1H), 7.47-7.43 (m, 2H), 7.37-7.35 (m, 3H), 6.88-6.84 (m, 2H), 5.06 (s, 2H), 2.46-2.42 (m, 1H), 2.26 (s, 3H), 1.34-1.21 (m, 6H).
Compound 10-1 (2.0 g, 7.45 mmol, 1.0 eq) was added to a 100-mL single-necked flask, and dry methanol (30 mL) and sodium borohydride (0.31 g, 8.20 mmol, 1.1 e.q) were added. The reaction mixture was reacted at 15° C. for 0.5 hours. After TLC detection showed that the reaction was completed, the reaction liquid was extracted with DCM (100 mL×2), and purified by column chromatography (eluent polarity:VPetroleum ether:VEthyl acetate=15:1 to 5:1) to obtain compound 10-2 as a yellow oil (2.00 g, yield: 100.0%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.65-7.61 (m, 1H), 7.48-7.44 (m, 2H), 7.39-7.35 (m, 3H), 6.88-6.84 (m, 2H), 5.08 (s, 2H), 3.65 (s, 1H), 2.46-2.42 (m, 2H), 1.91 (d, J=8.0 Hz, 3H), 1.34-1.21 (m, 6H).
Compound 10-2 (1.5 g, 5.55 mmol, 1.0 e.q) was added to a 100-mL single-necked flask, dry DCM (30 mL) and DMAP (0.678 g, 5.55 mmol, 1.0 e.q) were added, and then acetic anhydride (736.3 mg, 7.21 mmol, 1.3 e.q) was added. The reaction mixture was reacted at 15° C. for 0.5 hours. After TLC detection showed that the reaction was completed, the reaction mixture was extracted with DCM (100 mL×2). The reaction liquid was washed with hydrochloric acid (1 N), extracted, subjected to liquid separation and dried to obtain compound 10-3 as a yellow transparent oil (1.50 g, yield: 86.7%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.65-7.61 (m, 1H), 7.48-7.44 (m, 2H), 7.39-7.35 (m, 3H), 6.88-6.84 (m, 2H), 6.05-6.00 (m, 1H), 5.08(s, 2H), 2.46-2.42 (m, 1H), 2.12 (s, 3H), 1.93 (d, J=8.0 Hz, 3H), 1.34-1.21 (m, 6H).
Under the protection of a hydrogen balloon (15 psi), compound 10-3 (300 mg, 0.96 mmol, 1.0 eq) was added to dry THF (10 mL), and palladium on carbon (200 g, pd: 10%) was added. The reaction mixture was reacted at 15° C. in the hydrogen balloon (15 psi) for 5 hours. After TLC detection showed that the reaction was completed, the reaction compound was filtered through celite, washed, dried and concentrated to obtain a product, which was purified by column chromatography to obtain compound 10 as a light yellow oil (100 mg, yield: 46.9%).
1H NMR (400 MHz, CDCl3) δ7.64 (s, 1H), 7.19-7.18 (m, 2H), 6.94-6.90 (m, 1H), 6.05-6.00 (m, 1H), 3.40-3.33 (m, 1H), 2.08 (s, 3H), 1.65 (d, J=8.0 Hz, 3H), 1.24-1.21 (m, 6H).
Compound 3-5 (1.0 g, 4.85 mmol) was added to ethanethiol (361 mg, 5.81 mmol, 1.2 eq) under nitrogen protection, and hydrochloric acid (229 mg, 6.30 mmol, 1.3 eq) was added dropwise to the reaction liquid. The reaction system was stirred and reacted at 15° C. for 10 hours. After TLC (VHexane:VEA=5:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with H2O and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated saline and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=50:1 to 20:1) to obtain compound 11-1 as a yellow oil (700 mg, yield: 57.7%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.50 (d, J=8.0 Hz, 1H), 7.27-7.25 (m, 1H), 6.93-6.91 (m, 1H), 6.86-6.83 (m, 1H), 4.18-4.12 (m, 1H), 2.58-2.52 (m, 1H), 2.40-2.34 (m, 2H), 1.64 (d, J=8.0 Hz, 3H), 1.31-1.28 (m, 3H), 1.19-1.15 (m, 3H), 1.05-0.97 (m, 1H), 0.55-0.54 (m, 1H), 0.35-0.30 (m, 1H), 0.20-0.16 (m, 2H).
Under nitrogen protection, compound 11-1 (200 mg, 0.798 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (325 mg, 1.60 mmol, 2.0 eq) was slowly added to the reaction, and the reaction was conducted at 15° C. for 0.5 hours. After TLC (VHexane:VEA=1:1) detection showed that the raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:1) to obtain compound 11 as a light yellow oil (180.0 mg, yield: 82%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.38-7.33 (m, 1H), 7.12-7.10 (m, 2H), 6.99-6.96 (m, 1H), 4.59-4.55 (m, 1H), 2.94-2.88 (m, 2H), 2.58-2.54 (m, 1H), 1.83 (d, J=8.0 Hz, 3H), 1.34-1.32 (m, 3H), 1.30-1.27 (m, 3H), 1.05-1.01 (m, 1H), 0.58-0.56 (m, 1H), 0.42-0.40 (m, 1H), 0.23-0.15 (m, 2H).
Under nitrogen protection, compound 11-1 (200 mg, 0.798 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (146 mg, 0.718 mmol, 0.9 eq) was slowly added to the reaction, and the reaction was conducted at 0° C.-5° C. for 0.5 hours. After TLC (VHexane:VEA=1:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:1) to obtain compound 12 as a yellow oil (40.0 mg, yield: 18.8%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ9.79 (s, 1H), 7.32-7.29 (m, 1H), 6.86-6.79 (m, 2H), 3.82-3.80 (m, 1H), 2.64-2.56 (m, 2H), 2.40 -2.34 (m, 1H), 1.83 (d, J=8.0 Hz, 3H), 1.34-1.29 (m, 3H), 1.24-1.19 (m, 3H), 1.05-1.00 (m, 1H), 0.54-0.51 (m, 1H), 0.38-0.35 (m, 1H), 0.21-0.16 (m, 2H).
Compound 13-1 (10.0 g, 66.59 mmol, 1.0 eq) and NaOH (4.0 g, 99.88 mmol, 1.5 eq) were added to anhydrous DMF (150 mL), and the mixture was stirred at 15° C. for 30 minutes. Chlorobutene (a cis-trans mixture, 7.24 g, 79.91 mmol, 1.3 eq) was then added dropwise at 15° C. After the dropwise addition was completed, the mixture was slowly warmed to room temperature (15° C.) and reacted for 16 hours. The reaction liquid was slowly poured into ice water (500 mL), and extracted with n-hexane (400 mL×3). The organic phases were combined, and subjected to rotary evaporation to remove the solvent. The crude product was subjected to sample stirring, and purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=200:1) to obtain compound 13-2 as a yellow transparent oil (7.7 g, yield: 56.6%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.70-7.66 (m, 1H), 7.43-7.41 (m, 1H), 7.00-6.93 (m, 2H), 5.87-5.71 (m, 2H), 4.68-4.54 (m, 2H), 3.04-2.98 (m, 2H), 1.78 (d, J=8.0 Hz, 3H), 1.18-1.16 (m, 3H).
Compound 13-2 (7.70 g, 37.72 mmol, 1.0 eq) was added to a 50-mL single-necked flask. Under nitrogen protection, condensation was conducted with air reflux. The reaction mixture was heated to 210° C.-215° C. and reacted for 4.0 hours. After TLC detection showed that the reaction was completed, the reaction liquid was diluted with ethyl acetate, and subjected to sample stirring and column separation (eluent:VPetroleum ether:VEthyl acetate=100:1 to 50:1) to obtain compound 13-3 as a yellow oil (6.80 g, yield: 88.3%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ12.81 (s, 1H), 7.66-7.64 (m, 1H), 7.37-7.35 (m, 1H), 6.88-6.84 (m, 1H), 6.05-6.01 (m, 1H), 5.08-5.06 (m, 2H), 4.01-3.97 (m, 1H), 3.06-3.02 (m, 2H), 1.34(d, J=4.0 Hz, 3H), 1.24-1.20 (m, 3H).
Under nitrogen protection, a 100-mL three-necked flask was subjected to replacement 3-5 times, and DCM (20 mL) was added. The reaction system was cooled to −5° C.-0° C., and diethyl zinc (14.69 mL, 2.0 M, 29.37 mmol) was slowly added dropwise to the reaction liquid, with the addition completed in about 10 minutes. Then trifluoroacetic acid (3.35 g, 29.37 mmol) was added in an ice ethanol bath. After about 5 minutes, diiodomethane (10.49 g, 39.16 mmol) was dissolved in DCM (5 mL), and the mixture was added to the reaction liquid using a syringe. The system temperature was kept at −5° C.-0° C. After 60 minutes, compound 13-3 (2.00 g, 9.79 mmol) was dissolved in DCM (5 mL), and the mixture was slowly added dropwise to the reaction. After the dropwise addition was completed, the ice bath was removed. The reaction was stirred at −5° C.-0° C. for 3 hours, and then warmed to 15° C. and stirred for additional 48 hours. After 1H NMR and TLC (VHexane:VEA=10:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (200 mL), and washed with a saturated ammonium chloride solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent:VPetroleum ether:VEthyl acetate=100:1 to 50:1) to obtain compound 13-4 as a yellow oil (1.80 g, yield: 84.1%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ12.73 (s, 1H), 7.66-7.63 (m, 1H), 7.55-7.53 (m, 1H), 6.90 (t, J=8.0 Hz, 1H), 3.08-3.03 (m, 2H), 2.52-2.48 (m, 1H), 1.31 (d, J=4.0 Hz, 3H), 1.24-1.20 (m, 3H), 1.03-0.97 (m, 1H), 0.59-0.53 (m, 1H), 0.41-0.35 (m, 1H), 0.26-0.20 (m, 1H), 0.16-0.12 (m, 1H).
Compound 13-4 (3.30 g, 15.14 mmol, 1.0 eq) was added to dry methanol (50 mL), and then NaBH4 (624 mg, 16.49 mmol, 1.1 eq) was added. The mixture was reacted at room temperature for 0.5 hours. The reaction liquid was poured into a saturated ammonium chloride aqueous solution (10 mL), and extracted with DCM (200 mL×3). The organic phases were combined, washed with saturated saline, and dried over anhydrous sodium sulphate. The crude product was purified by column chromatography (eluent polarity:VPE:VEA=100:1 to 20:1) to obtain compound 13-5 as a yellow oil (2.00 g, yield: 60.1%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ8.10 (s, 1H), 7.25-7.23 (m, 1H), 6.82-6.79 (m, 2H), 4.75 (t, J=8.0 Hz, 1H), 2.52-2.45 (m, 2H), 1.98 -1.84 (m, 2H), 1.30 (dd, J=8.0 Hz, 2.0 Hz, 3H), 1.05-1.01 (m, 1H), 0.98 (t, J=8.0 Hz, 3H), 0.56-0.52 (m, 1H), 0.41-0.35 (m, 1H), 0.22-0.14 (m, 2H).
Compound 13-5 (400 mg, 1.82 mmol) was added to acetonitrile (10 mL), and methanethiol (1.05 g, 10% propylene glycol solution, 2.18 mmol, 1.2 eq) was added under nitrogen protection. Then hydrochloric acid (72.82 mg, 2.00 mmol, 1.1 eq) was added dropwise to the reaction liquid. The reaction system was stirred and reacted at 25° C. for 6 hours. After TLC (VHexane:VEA=5:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with water and extracted with ethyl acetate (30 mL×3) . The organic phases were combined, washed with saturated saline and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:0 to 100:1) to obtain compound 13 as a yellow oil (110 mg, yield: 24.2%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.43 (d, J=4.0 Hz, 1H), 7.26-7.25 (m, 1H), 6.87-6.81 (m, 2H), 3.80 (t, J=8.0 Hz, 1H), 2.58-2.53 (m, 1H), 2.00-1.92 (m, 2H), 1.88 (d, J=2.0 Hz, 3H), 1.30 (dd, J=8.0 Hz, 4.0 Hz, 3H), 1.03-1.00 (m, 1H), 0.98 (t, J=8.0 Hz, 3H), 0.54-0.52 (m, 1H), 0.38-0.35 (m, 1H), 0.20-0.14 (m, 2H).
Under nitrogen protection, compound 13 (100 mg, 0.439 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (164 mg, 0.878 mmol, 2.0 eq) was slowly added to the reaction, and the reaction was conducted at 15° C. for 0.5 hours. After TLC (VHexane:VEA=2:1) detection showed that the raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=10:1) to obtain compound 14 as a yellow oil (85.0 mg, yield: 75.2%).
Characterization data: 1NMR (400 MHz, CDCl3) δ7.35-7.32 (m, 1H), 7.10-7.05 (m, 1H), 7.00-6.96 (m, 1H), 4.48-4.28 (m, 1H), 2.73 (s, 3H), 2.62-2.54 (m, 1H), 2.48-2.42 (m, 1H), 2.32-2.18 (m, 1H), 1.30 (dd, J=8.0, 4.0 Hz, 3H), 1.04-0.99 (m, 1H), 0.95 (t, J=8.0 Hz, 3H), 0.59-0.54 (m, 1H), 0.44-0.42 (m, 1H), 0.24-0.12 (m, 2H).
Compound 13-5 (200 mg, 0.91 mmol) was added to acetonitrile (5 mL), and isopropylthiol (82.97 mg, 1.09 mmol, 1.2 eq) was added under nitrogen protection. Then hydrochloric acid (36.41 mg, 0.998 mmol, 1.1 eq) was added dropwise to the reaction liquid. The reaction system was stirred and reacted at 25° C. for 6 hours. After TLC (VHexane:VEA=10:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with H2O and extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed with saturated saline and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:0 to 100:1) to obtain compound 15-1 as a yellow oil (250 mg, yield: 98.8%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.78 (d, J=8.0 Hz, 1H), 7.26-7.24 (m, 1H), 6.85-6.79 (m, 2H), 3.91 (q, J=12.0, 8.0, 4.0 Hz, 1H), 2.62-2.52 (m, 2H), 1.99-1.84 (m, 2H), 1.30-1.27 (m, 3H), 1.22 (dd, J=8.0, 4.0 Hz, 3H), 1.16 (dd, J=8.0, 4.0 Hz, 3H), 1.05-0.98 (m, 1H), 0.96 (dt, J=8.0 Hz, 3H), 0.56-0.49 (m, 1H), 0.39-0.31 (m, 1H), 0.21-0.13 (m, 2H).
Under nitrogen protection, compound 15-1 (200 mg, 0.719 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (291 mg, 1.438 mmol, 2.0 eq) was slowly added to the reaction, and the reaction was conducted at 15° C. for 0.5 hours. After TLC (VHexane:VEA=3:1) detection showed that the raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=15:1) to obtain compound 15 as a white solid (160 mg, yield: 71.7%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ8.08 (d, J=8.0 Hz, 1H), 7.33-7.31 (m, 1H), 7.00-6.79 (m, 2H), 4.68-4.52 (m, 1H), 3.12-3.06 (m, 1H), 2.63-2.55 (m, 1H), 2.46-2.29 (m, 2H), 1.34-1.24 (m, 9H), 1.02-1.00 (m, 1H), 0.89 (t, J=8.0 Hz, 3H), 0.57-0.53 (m, 1H), 0.42-0.35 (m, 1H), 0.23-0.12 (m, 2H).
Compound 16-1 (4.0 g, 29.37 mmol, 1.0 eq) was added to a 100-mL single-necked flask, dry DCM (60 mL) and cyclopropanecarbonyl chloride (3.68 g, 35.24 mmol, 1.2 eq) were added, and then titanium tetrachloride (6.69 g, 35.24 mmol, 1.2 e.q) was added slowly to the reaction. The reaction mixture was kept at −30° C. and reacted for 2 hours under nitrogen protection. After TLC detection showed that the reaction was completed, the reaction liquid was diluted with ethyl acetate, and subjected to sample stirring and column separation (eluent:VPetroleum ether:VEthyl acetate=100:1 to 50:1) to obtain compound 16-2 as a yellow oil (200 mg, yield: 3.3%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ12.96 (s, 1H), 7.86 (d, J=8.0 Hz, 1H), 7.45 (d, J=8.0 Hz, 1H), 6.93 (t, J=6.0 Hz, 1H), 3.4-3.32 (m, 1H), 2.75-2.68 (m, 1H), 1.32-1.27 (m, 1H), 1.26 (d, J=6.0 Hz, 6H), 1.11-1.06 (m, 1H), 1.05-0.99 (m, 2H).
Compound 16-2 (200 mg, 0.979 mmol, 1.0 eq) was added to a 100-mL single-necked flask, and dry methanol (8 mL) and sodium borohydride (40.75 mg, 1.08 mmol, 1.1 e.q) were added. The reaction mixture was reacted at 15° C. for 0.5 hours. After TLC detection showed that the reaction was completed, the reaction liquid was extracted with DCM (30 mL×2), and purified by column chromatography (eluent polarity:VPetroleum ether:VEthyl acetate=15:1 to 5:1) to obtain compound 16-3 as a reddish-brown oil (200 mg, yield: 99.0%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ8.29 (s, 1H), 7.19 (, J=8.0 Hz, 1H), 6.96 (d, J=8.0 Hz, 1H), 6.85 (t, J=8.0 Hz, 1H), 4.14 (d, J=8.0 Hz, 1H), 3.39 (m, 1H), 2.47 (s, 1H), 1.52-1.41 (m, 1H), 1.27 (d, J=6.0 Hz, 6H), 0.74-0.66 (m, 2H), 0.48-0.41 (m, 2H).
Compound 16-3 (200 mg, 0.97 mmol) was added to methanethiol (0.606 g, 10% propylene glycol solution, 1.26 mmol, 1.3 eq) under nitrogen protection, and concentrated hydrochloric acid (42.4 mg, 1.16 mmol, 1.2 eq) was added dropwise to the reaction liquid. The reaction system was stirred and reacted at 25° C. for 4 hours. After TLC (VHexane:VEA=5:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with H2O and extracted with ethyl acetate (10 mL×3). The organic phases were combined, washed with saturated saline and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=50:1 to 20:1) to obtain compound 16-4 as a yellow oil (50 mg, yield: 21.8%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.56 (s, 1H), 7.20-7.19 (m, 1H), 6.98-6.95 (m, 1H), 6.89-6.85 (m, 1H), 3.42-3.38 (m, 1H), 3.22-3.19 (m, 1H), 1.94 (s, 3H), 1.48-1.41 (m, 1H), 1.28-1.25 (m, 6H), 0.79-0.73 (m, 1H), 0.65-0.60 (m, 1H), 0.46-0.41 (m, 1H), 0.36-0.30 (m, 1H).
Under nitrogen protection, compound 16-4 (50 mg, 0.210 mmol) was dissolved in DCM (8 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (84.9 mg, 0.42 mmol, 2.0 eq) was slowly added to the reaction, and the reaction was stirred at 15° C. for 0.5 hours. After TLC (VHexane:VEA=2:1) detection showed that the raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:1) to obtain compound 16 as a yellow solid (20.1 mg, yield: 35.1%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.24-7.22 (m, 2H), 7.03 (t, J=8.0 Hz, 1H), 6.85-6.82 (m, 1H), 3.72 (d, J=8.0 Hz, 1H), 3.37-3.31 (m, 1H), 2.92 (s, 3H), 1.75-1.66 (m, 1H), 1.25-1.22 (m, 6H), 1.06-1.02 (m, 1H), 0.78-0.74 (m, 2H), 0.25-0.20 (m, 1H).
Compound 17-1 (5.0 g, 33.29 mmol, 1.0 eq) was added to a 200-mL three-necked flask under nitrogen protection, dry THF (60 mL) was added, and ethylmagnesium bromide (19.98 mL, 2.0 M in THF, 39.95 mmol, 1.2 e.q) was added slowly to the reaction at −15° C. The reaction mixture was reacted at −15° C. for 2 hours. After TLC detection showed that the reaction was completed, the reaction liquid was diluted with ethyl acetate, and subjected to sample stirring and column separation (eluent:VPetroleum ether:VEthyl acetate=100:1 to 50:1) to obtain compound 17-2 as a yellow oil (4.0 g, yield: 74.6%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.63 (s, 1H), 7.24-7.16 (m, 2H), 6.97-6.94 (m, 1H), 6.76-6.73 (m, 1H), 6.06-6.00 (m, 1H), 2.06-2.00 (m, 5H), 1.06-1.02 (m, 3H).
Under hydrogen (15 Psi) protection, compound 17-2 (4.0 g, 24.66 mmol, 1.0 eq) was added to a 250-mL single-necked flask, dry methanol (60 mL) was added, and palladium on carbon (800 mg, Pd: 10%) was added to the reaction at 15° C. The reaction mixture was reacted in hydrogen (15 Psi) at 15° C. for 12 hours. After TLC detection showed that the reaction was completed, the reaction liquid was filtered through celite, washed with methanol, dried and concentrated to obtain compound 17-3 as a grey oil (4.0 g, yield: 95.0%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.67 (s, 1H), 7.24-7.16 (m, 2H), 6.97-6.94 (m, 1H), 6.76-6.73 (m, 1H), 2.45-2.42 (m, 1H), 2.06-2.00 (m, 4H), 1.06-1.02 (m, 6H).
Compound 17-3 (4.0 g, 24.35 mmol, 1.0 eq) was added to a 250-mL single-necked flask under nitrogen protection, dry DCM (60 mL) was added, and triethylamine (3.7 g, 36.53 mmol, 1.5 e.q.) and acetic anhydride (2.98 g, 29.22 mmol, 1.2 e.q.) were added to the reaction at 15° C. The reaction mixture was reacted at 15° C. for 12 hours. After TLC detection showed that the reaction was completed, the reaction liquid was neutralized with dilute hydrochloric acid to pH=6-7, washed with saturated saline, dried over anhydrous sodium sulphate, and concentrated to obtain compound 17-4 as a colourless oil (4.5 g, yield: 89.6%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.24-7.16 (m, 2H), 6.97-6.94 (m, 1H), 6.76-6.73 (m, 1H), 2.45-2.42 (m, 1H), 2.36 (s, 3H), 2.06-2.00 (m, 4H), 1.06-1.02 (m, 6H).
Compound 17-4 (4.5 g, 21.81 mmol, 1.0 eq) was added to a 100-mL single-necked flask under nitrogen protection, dry DCM (60 mL) was added, and aluminium trichloride (3.78 g, 28.36 mmol, 1.3 e.q.) was added in portions to the reaction at 15° C. The reaction mixture was heated to 140° C. and reacted for 4 hours. After TLC detection showed that the reaction was completed, the reaction liquid was extracted with DCM. The organic phase was washed with saturated saline, dried and concentrated. The crude product was purified by column chromatography to obtain compound 17-5 as a yellow oil (2.26 g, yield: 50.1%).
1H NMR (400 MHz, CDCl3) δ7.37 (s, 1H), 7.24-7.16 (m, 1H), 6.97-6.94 (m, 1H), 6.76-6.73 (m, 1H), 2.45-2.42 (m, 1H), 2.18 (s, 3H), 2.06-2.02 (m, 4H), 1.08-1.04 (m, 6H).
Compound 17-5 (2.0 g, 9.70 mmol, 1.0 eq) was added to dry methanol (30 mL) under nitrogen protection, and sodium borohydride (0.440 g, 11.61 mmol, 1.2 e.q.) was added in portions to the reaction at 15° C. The reaction mixture was reacted at 20° C. for 1 hour. After TLC detection showed that the reaction was completed, the reaction liquid was extracted with DCM. The organic phase was washed with saturated saline, dried and concentrated to obtain compound 17-6 as a yellow oil (2.01 g, yield: 99.0%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.37 (s, 1H), 7.24-7.17 (m, 1H), 6.97-6.94 (m, 1H), 6.76-6.73 (m, 1H), 2.54 (s, 1H), 2.45-2.42 (m, 1H), 2.06-1.98 (m, 4H), 1.91 (s, 3H), 1.24-1.17 (m, 6H).
Compound 17-6 (2.0 g, 9.60 mmol) was added to methanethiol (5.54 g, 10% propylene glycol solution, 11.52 mmol, 1.2 eq) under nitrogen protection, and hydrochloric acid (420 mg, 11.52 mmol, 1.2 eq) was added dropwise to the reaction liquid. The reaction system was stirred and reacted at 15° C. for 12 hours. After TLC (VHexane:VEA=5:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with H2O and extracted with ethyl acetate (100 mL×3). The organic phases were combined, washed with saturated saline and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=80:1 to 25:1) to obtain compound 17-7 as a light yellow oil (1.1 g, yield: 48.1%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.63 (s, 1H), 7.26-7.21 (m, 1H), 7.12-7.06 (m, 1H), 6.96-6.86 (m, 1H), 4.05-4.00 (m, 1H), 2.45-2.42 (m, 1H), 2.06-1.98 (m, 4H), 1.94 (s, 3H), 1.63 (d, J=8.0 Hz, 3H), 1.21-1.14 (m, 6H).
Under nitrogen protection, compound 17-7 (400 mg, 1.68 mmol) was dissolved in DCM (12 mL), and the system temperature was kept at 5° C.-0° C. Meta-chloroperoxybenzoic acid (681.3 mg, 3.36 mmol, 2.0 eq) was slowly added to the reaction, and the reaction was conducted at 15° C. for 0.5 hours. After TLC (VHexane:VEA=2:1) detection showed that the raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (30 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:1) to obtain compound 17 as a yellow oil (100 mg, yield: 22.0%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.65 (s, 1H), 7.26-7.18 (m, 1H), 7.12-7.06 (m, 1H), 6.96-6.86 (m, 1H), 4.62-4.58 (m, 1H), 2.78 (s, 3H), 2.35-2.30 (m, 1H), 2.06-1.98 (m, 4H), 1.83 (d, J=8.0 Hz, 3H), 1.21-1.14 (m, 6H).
Compound 13-5 (200 mg, 0.91 mmol) was added to acetonitrile (5 mL), and ethanethiol (67.7 mg, 1.09 mmol, 1.2 eq) was added under nitrogen protection. Then hydrochloric acid (36.41 mg, 0.998 mmol, 1.1 eq) was added dropwise to the reaction liquid. The reaction system was stirred and reacted at 25° C. for 6 hours. After TLC (VHexane:VEA=10:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with H2O and extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed with saturated saline and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=1:0 to 100:1) to obtain compound 18-1 as a yellow oil (210 mg, yield: 87.5%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.58 (d, J=4.0 Hz, 1H), 7.26-7.24 (m, 1H), 6.86-6.80 (m, 2H), 3.90 (d, J=8.0 Hz, 1H), 2.58-2.53 (m, 1H), 2.34-2.28 (m, 2H), 1.99-1.88 (m, 2H), 1.30 (dd, J=8.0 , 4.0 Hz, 3H), 1.16-1.12 (m, 3H), 1.04-0.98 (m, 1H), 0.96 (t, J=8.0 Hz, 3H), 0.55-0.51 (m, 1H), 0.39-0.33 (m, 1H), 0.21-0.13 (m, 2H).
Under nitrogen protection, compound 18-1 (200 mg, 0.820 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (306 mg, 1.64 mmol, 2.0 eq) was slowly added to the reaction, and the reaction was conducted at 15° C. for 0.5 hours. After TLC (VHexane:VEA=2:1) detection showed that the raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=15:1) to obtain compound 18 as a yellow oil (200 mg, yield: 89.3%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.34-7.31 (m, 1H), 7.03-7.00 (m, 1H), 6.97-6.93 (m, 1H), 4.47-4.05 (m, 1H), 2.90-2.83 (m, 2H), 2.64-2.53 (m, 1H), 2.46-2.28 (m, 2H), 1.31-1.27 (m, 6H), 1.05-0.98 (m, 1H), 0.92 (t, J=8.0 Hz, 3H), 0.59-0.52 (m, 1H), 0.43-0.40 (m, 1H), 0.23-0.10 (m, 2H).
Compound 3-5 (1.0 g, 4.85 mmol) was added to acetonitrile (10 mL), and trifluoroethylthiol (674 mg, 5.81 mmol, 1.2 eq) was added under nitrogen protection. Then hydrochloric acid (229 mg, 6.30 mmol, 1.3 eq) was added dropwise to the reaction liquid. The reaction system was stirred and reacted at 15° C. for 10 hours. After TLC (VHexane:VEA=5:1) detection showed that a majority of raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with H2O and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated saline and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=50:1 to 20:1) to obtain compound 19-1 as a yellow oil (800 mg, yield: 65.7%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.50 (d, J=8.0 Hz, 1H), 7.27-7.25 (m, 1H), 6.93-6.91 (m, 1H), 6.86-6.83 (m, 1H), 4.16-4.08 (m, 1H), 2.93-2.82 (m, 2H), 2.58-2.52 (m, 1H), 1.64 (d, J=8.0 Hz, 3H), 1.31-1.28 (m, 3H), 1.05-0.97 (m, 1H), 0.59-0.54 (m, 1H), 0.38-0.33 (m, 1H), 0.20-0.16 (m, 2H).
Under nitrogen protection, compound 19-1 (200 mg, 0.657 mmol) was dissolved in DCM (10 mL), and the system temperature was kept at −5° C.-0° C. Meta-chloroperoxybenzoic acid (265.9 mg, 1.314 mmol, 2.0 eq) was slowly added to the reaction, and the reaction was conducted at 15° C. for 0.5 hours. After TLC (VHexane:VEA=1:1) detection showed that the raw materials were consumed, the reaction was stopped. The reaction liquid was diluted with dichloromethane (10 mL), and washed with a saturated sodium bicarbonate solution. The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulphate, filtered and concentrated. The crude product was purified by column chromatography (eluent polarity:Vn-Hexane:VEthyl acetate=15:1) to obtain compound 19 as a colourless oil (185.0 mg, yield: 83.7%).
Characterization data: 1H NMR (400 MHz, CDCl3) δ7.63-7.59 (m, 1H), 7.32-7.27 (m, 2H), 7.10-7.06 (m, 1H), 4.75-4.68 (m, 1H), 3.94-3.88 (m, 2H), 2.56-2.54 (m, 1H), 1.92(d, J=8.0 Hz, 3H), 1.35-1.24 (m, 3H), 1.05-1.00 (m, 1H), 0.59-0.54 (m, 1H), 0.42-0.40 (m, 1H), 0.28-0.16 (m, 2H).
Characterization data: 1H NMR (400 MHz, Chloroform-d) δ7.36 (dd, J=7.6, 1.7 Hz, 1H), 7.12 (dd, J=7.7, 1.7 Hz, 1H), 7.10 (s, 1H), 7.00 (t, J=7.7 Hz, 1H), 4.60 (q, J=7.3 Hz, 1H), 2.93 (q, J=7.5 Hz, 2H), 2.65-2.53 (m, 1H), 1.85 (d, J=7.2 Hz, 3H), 1.38-1.28 (m, 6H), 1.08-0.99 (m, 1H), 0.62-0.56 (m, 1H), 0.49-0.38 (m, 1H), 0.28-0.22 (m, 1H), 0.20-0.14 (m, 1H).
Characterization data: 1H NMR (400 MHz, Chloroform-d) δ7.36 (d, J=7.6 Hz, 1H), 7.13 (d, J=8.0 Hz, 1H),7.11 (s, 1H), 7.01 (t, J=7.7 Hz, 1H), 4.60 (q, J=7.3 Hz, 1H), 2.94 (q, J=7.4 Hz, 2H), 2.69-2.52 (m, 1H), 1.85 (d, J=7.4 Hz, 3H), 1.36-1.30 (m, 6H), 1.09-1.00 (m, 1H), 0.62-0.56 (m, 1H), 0.49-0.37 (m, 1H), 0.26-0.21 (m, 1H), 0.20-0.15 (m, 1H).
Characterization data: 1H NMR (400 MHz, Chloroform-d) δ7.37 (dd, J=7.6, 1.7 Hz, 1H), 7.13 (dd, J=7.7, 1.7 Hz, 1H),7.11 (s, 1H), 7.00 (t, J=7.7 Hz, 1H), 4.60 (q, J=7.3 Hz, 1H), 2.93 (q, J=7.5 Hz, 2H), 2.74-2.46 (m, 1H), 1.85 (d, J=7.3 Hz, 3H), 1.41-1.25 (m, 6H), 1.10-0.97 (m, 1H), 0.62-0.56 (m, 1H), 0.47-0.40 (m, 1H), 0.28-0.22 (m, 1H), 0.20-0.14 (m, 1H).
Characterization data: 1H NMR (400 MHz, Chloroform-d) δ7.36 (dd, J=7.7, 1.7 Hz, 1H), 7.13 (dd, J=7.8, 1.6 Hz, 1H), 7.11 (s, 1H), 7.00 (t, J=7.7 Hz, 1H), 4.60 (q, J=7.3 Hz, 1H), 2.93 (q, J=7.5 Hz, 2H), 2.66-2.54 (m, 1H), 1.85 (d, J=7.3 Hz, 3H), 1.39-1.23 (m, 6H), 1.08-1.00 (m, 1H), 0.62-0.56 (m, 1H), 0.47-0.41 (m, 1H), 0.27-0.21 (m, 1H), 0.20-0.14 (m, 1H).
Preparation method of Examples 20-23: Chiral isomer 2-(1-cyclopropylethyl)-6-(1-(ethylsulfonyl)ethyl)phenol (compound 11) (1.5 g, 5.3 mmol) was subjected to chiral resolution by an HPLC method with preparation equipment and a chiral column.
Separation conditions: chiral column: CHIRALCEL OJ-H, mobile phase: Vn-Hexane:VEthanol=90:10, flow rate: 25 mL/min, UV=214 nm, and column temperature: 35° C.
Components with retention times of 6.908 min, 10.044 min and 11.829 min were collected respectively, and concentrated under reduced pressure,
SPF-grade ICR mice, each weighing 18-22 g, half male and half female, were used. A well-established mouse anesthesia model was used to study the general anesthesia effect of the compound of the present invention. The compounds were formulated with solvents of 10% DMSO, 15% solutol (HS15) and 75% saline to the desired concentration, for further use. After adapting to the experimental environment, the experimental animals were fasted with water available for 12 h, and were administered via intravenous injection at an administration volume of 10 mL/kg. Then the anesthesia induction time (the time period from drug administration to disappearance of the righting reflex) and the anesthesia maintenance time (the time period from disappearance of the righting reflex to recovery of the righting reflex) were recorded. Indicators including the median effective dose (ED50), median lethal dose (LD50), therapeutic index (TI, i.e. LD50/ED50), anesthesia induction time, anesthesia maintenance time and maximum tolerated dose were used to evaluate the effect and safety of anesthesia. Among them, control group 1 (propofol), control group 2 (ciprofol, a racemate), and control group 3 (ciprofol, R-configuration) respectively had the following structural formulas. The specific experimental results were shown in Table 1.
Experimental conclusion: compared with control group 1, control group 2 and control group 3, the compound of the present invention showed higher therapeutic index and safety coefficient, and had a broader therapeutic window. The compound of the present invention had lower ED50 value, indicating that the compound had a lower minimum effective dose and higher activity. The compound of the present invention showed a low free concentration in the aqueous phase of corresponding formulations, and was expected to have an effect of avoiding pain on injection.
SD rats, half male and half female, were fasted with water available for 12 h, and were administered via intravenous injection at an administration volume of 10 mL/kg. Then the anesthesia induction time (the time period from drug administration to disappearance of the righting reflex), the anesthesia maintenance time (the time period from disappearance of the righting reflex to recovery of the righting reflex and revival) and the anesthesia recovery time (the time period from revival to complete recovery) were recorded. Indicators including the median effective dose (ED50), median lethal dose (LD50), therapeutic index (TI, i.e. LD50/ED50), anesthesia induction time, anesthesia maintenance time and maximum tolerated dose were used to evaluate the effect and safety of anesthesia.
Experimental conclusion: compared with control group 3, compound 20 of the present invention showed higher therapeutic index and safety coefficient, and had a broader therapeutic window. Compound 20 of the present invention had lower ED50 value, indicating that the compound had a lower minimum effective dose than that of control group 3 and higher activity.
Gamma-aminobutyric acid (GABA) was an important inhibitory amino acid neurotransmitter in the central nervous system, and functioned by binding to GABA receptors. The GABA receptors were divided into three subtypes: GABAA, GABAB and GABAC, among which the GABAA receptor was the most important. The GABAA receptor was an anion-selective ion channel that could enhance chloride ion permeability and thus reduce neuronal excitability. The GABAA receptor was involved in regulating general anesthesia and was also closely related to diseases such as neurological and psychiatric disorders, e.g., depression, insomnia, anxiety and epilepsy. A whole-cell patch-clamp technique was used to study the allosteric regulation effect of the compound of the present invention on the GABAA (α1β2γ2) receptor.
HEK293 cell lines stably expressing the GABAA (α1β2γ2) receptor were used. The GABAA (α1β2γ2) receptor gene information was as follows: GABA-α1: NM_000806; GABA-β2: NM_021911; GABA-γ2: NM_198904. The voltage stimulus of GABA receptor current recorded by a whole-cell patch-clamp technique was as follows: when whole-cell sealing was formed, the cell membrane voltage was clamped at −70 mV. The peak value of current was recorded after sequentially spraying test compounds from low concentration to high concentration and 100 μM GABA onto the cell surface in a Gap-free mode. The mode of administration of test compounds was as follows: for each concentration, the test compound was administered 1-2 times; the cells were washed with extracellular fluid for 1 min before detection was performed on the test compound at another concentration; and finally, 100 μM GABA was given as the control. The experimental data was collected by an EPC-10 amplifier (HEKA) and stored in PatchMaster (HEKA) software. A microelectrode puller was used to pull capillary glass tubes into a recording electrode. A microelectrode manipulator was manipulated under an inverted microscope to contact the recording electrode with cells, and negative pressure suction was applied to form a GΩ seal. After the GΩ seal was formed, rapid capacitance compensation (pF) was conducted, and then negative pressure was continued to break cell membranes, forming a whole-cell recording mode. Then slow capacitance compensation was conducted, and the membrane capacitance (pF) and series resistance were recorded. No electric leakage compensation was provided. The cover glass spread with cells was placed in a recording bath of the inverted microscope, and a working solution of the test compound and the extracellular fluid without the compound sequentially flowed through the recording bath from low concentration to high concentration by gravity perfusion to act on the cells. During the recording, a vacuum pump was used for liquid exchange. Multiple cells were independently and repeatedly detected. All electrophysiological experiments were conducted at room temperature.
The allosteric regulation effect of the compound of the present invention on the GABAA (α1β2γ2) receptor was detected in three independent repeated experiments, and the semi-activated concentration (EC50) of the sample on the GABAA (α1β2γ2) receptor was calculated by means of fitting. The experimental results were as follows:
Experimental conclusion: the semi-activated concentration (EC50) of compound 20 of the present invention having allosteric regulation on GABAA (α1β2γ2) was 1.556 μM, and the EC50 of control group 3 was 10.40 μM. The results indicated that compared with control group 3, compound 20 of the present invention had a stronger allosteric regulation effect on GABAA (α1β2γ2), and could exert anesthesia effects by agonizing GABAA (α1β2γ2).
Twelve male SD rats (200-300 g) were randomized into 2 groups (n=6): control group 3 and compound 20. The rats were administered via intravenous injection at 1 mg/kg (control group 3) and 1 mg/kg (compound 20), respectively, with an administration volume of 5 mL/kg and a vehicle of 5% DMSO+10% solutol (HS15)+85% Saline. Blood samples were taken before administration and at 2 min, 4 min, 8 min, 12 min, 15 min, 30 min, 1 h, 1.5 h and 2 h after administration. Plasma samples were taken by centrifugation, and stored in a −80° C. refrigerator until LC-MS analysis to be tested. After sample processing, quantitative analysis of substances in plasma was conducted using LCMS/MS to detect the concentration of prototype compounds in the plasma. The plasma concentration/time curve obtained in this way was used to calculate pharmacokinetic parameters by a validated pharmacokinetic computer program. The experimental results were shown in Table 4.
Experimental conclusion: the pharmacokinetic properties of compound 20 of the present invention were similar to those of control group 3. Metabolism clearance in rats was faster, which was consistent with the anesthesia effect. After anesthesia, the rats could wake up quickly, while avoiding toxicity accumulation in the bodies, suggesting that the compound had good pharmacokinetic properties.
Six male Beagle dogs (6-10 kg) were randomized into 2 groups (n=3): control group 3 and compound 20. The rats were administered via intravenous injection at 0.5 mg/kg (control group 3) and 0.5 mg/kg (compound 20), respectively, with an administration volume of 1 mL/kg and a vehicle of 5% DMSO+10% solutol (HS15)+85% Saline. Blood samples were taken before administration and at 2 min, 5 min, 10 min, 20 min, 30 min, 1 h, 1.5 h, 2 h and 3 h after administration. Plasma samples were taken by centrifugation, and stored in a −80° C. refrigerator until LC-MS analysis to be tested. After sample processing, quantitative analysis of substances in plasma was conducted using LCMS/MS to detect the concentration of prototype compounds in the plasma. The plasma concentration/time curve obtained in this way was used to calculate pharmacokinetic parameters by a validated pharmacokinetic computer program. The experimental results were shown in Table 5.
Experimental conclusion: the pharmacokinetic properties of compound 20 of the present invention were similar to those of control group 3. Metabolism clearance in Beagle dogs was faster, and toxicity accumulation in the bodies was avoided, suggesting that the compound had good pharmacokinetic properties.
The experiment used an emka PACK 4G telemetry system to detect the effects of intravenous injection of control group 3 and compound 20 on electrocardiogram, blood pressure and body temperature in conscious Beagle dogs, providing reference information for evaluating the safety of clinical use.
Four Beagle dogs were subjected to crossover administration, with a vehicle control (5% DMSO+10% solutol+85% Saline), control group 3 (2 mg/kg), compound 20 (0.5 mg/kg) and compound 20 (1 mg/kg) for each intravenous injection, with an administration volume of 2 mL/kg. Observations were respectively conducted before administration and within 1 h after administration on the day of administration. Anal temperature was measured before administration and at 30 min, 1 h, 2 h and 3 h after administration during each administration period; and electrocardiogram parameters were continuously collected from 0.5 h before administration to 3 h after administration during each administration period. Blood pressure was collected at 15 min after collection was started, and was collected at 2 min, 5 min, 10 min, 20 min, 30 min, 1 h, 2 h and 3 h after administration. The electrocardiogram, blood pressure data, body temperature, and clinical observation results on the day of administration at various time points including before administration (at 30 min after collection was started) and at 2 min, 5 min, 10 min, 20 min, 30 min, 1 h, 2 h and 3 h after administration were analysed and evaluated. The experimental results were shown in
Experimental conclusion: the experimental results showed a slow down in heart rate of animals within 1 h after administration of control group 3 and compound 20, and a recovery in heart rate after 1 h. After administration, although all indicators of blood pressure of animals in each group decreased, the amplitude was not significant, among which the systolic pressure decreased unapparently, and was still within the normal range. After administration, there were no abnormalities in the body temperature indicator of animals in each group. The results indicated that compound 20 of the present invention had little effect on the heart rate, blood pressure and body temperature of Beagle dogs, and therefore, it was expected that compound 20 of the present invention had good safety.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202110117767.2 | Jan 2021 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2022/073472 | 1/24/2022 | WO |
