A deuterated carbamate compound and its application in medicine

Abstract

The present invention relates to a deuterated carbamate compound, or a stereoisomer and a deuterated product thereof, and application thereof in medicine.

Description

Technical Field

[0001] The present invention relates to a deuterated carbamate compound, or a stereoisomer and a deuterated product thereof, and application thereof in medicine. Background Art

[0002] Epilepsy, commonly known as "epilepsy" or "epilepsy", is a chronic disease in which brain neurons suddenly discharge abnormally, leading to transient brain dysfunction. Currently, the first-line anti-epileptic drugs are mainly carbamazepine, sodium valproate, levetiracetam, oxcarbazepine, and lamotrigine, but they all have certain side effects, including dizziness, headache, mental tension, mental depression, nausea, etc., and about 30% of patients still cannot effectively control epileptic seizures after taking the drugs. Therefore, there is an urgent need to develop safer and more effective anti-epileptic drugs. Summary of the invention

[0003] The purpose of the present invention is to provide a deuterated carbamate compound or its stereoisomer, deuterated product, a pharmaceutical composition thereof, and use thereof in the preparation of anti-epileptic drugs.

[0004] One or more embodiments of the present application provide a compound or a stereoisomer or deuterated product thereof. The compound of the present application is selected from:

[0005]

[0006] in:

[0007] R1, R2 and R3 are each independently selected from H or D, and R1, R2 and R3 are not all H;

[0008] R4 is selected from H, D, halogen, C1-C6 alkyl or haloalkyl of C1-C6 alkyl;

[0009] R5 is selected from halogen or haloalkyl;

[0010] m is selected from 0, 1, 2, 3 or 4.

[0011] One or more embodiments of the present application provide a compound or a stereoisomer or deuterated product thereof. The compound of the present application is selected from, but not limited to, the following structures:

[0012]

[0013] One or more embodiments of the present invention provide a pharmaceutical composition comprising:

[0014] (1) The compounds of the present invention or their stereoisomers or deuterated derivatives;

[0015] (2) optionally one or more other active ingredients; and

[0016] (3) Pharmaceutically acceptable carriers and / or excipients.

[0017] One or more embodiments of the present invention provide use of the compound of the present invention or its stereoisomer, deuterated substance, or the pharmaceutical composition of the present invention in the preparation of an anti-epileptic drug.

[0018] Unless stated to the contrary, the terms used in the specification and claims have the following meanings.

[0019] The carbon, hydrogen, oxygen, sulfur, nitrogen or F, Cl, Br, I involved in the groups and compounds of the present invention include their isotopes, and the carbon, hydrogen, oxygen, sulfur or nitrogen involved in the groups and compounds of the present invention are optionally further replaced by one or more of their corresponding isotopes, wherein the carbon isotopes include 12 C. 13 C and 14 C, hydrogen isotopes include protium (H), deuterium (D, also called heavy hydrogen), tritium (T, also called super tritium), oxygen isotopes include 16 O. 17 O and 18 O, sulfur isotopes include 32 S. 33 S. 34 S and 36 S, isotopes of nitrogen include 14 N and 15 N, fluorine isotopes include 17 F and 19 F, chlorine isotopes include 35 Cl and 37 Isotopes of Cl and bromine include 79 Br and 81 Br.

[0020] "Pharmaceutical composition" refers to a mixture of one or more compounds described herein, their pharmaceutically acceptable salts or prodrugs and other chemical components, wherein "other chemical components" refers to pharmaceutically acceptable carriers, excipients and / or one or more other therapeutic agents.

[0021] "Carrier" refers to a material that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

[0022] "Excipient" refers to an inert substance added to a pharmaceutical composition to facilitate administration of a compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, and disintegrants.

[0023] "Stereoisomers" refer to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and conformational isomers.

[0024] "Optional" or "optionally" or "selective" or "selectively" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs and instances where it does not occur. For example, "heterocyclyl optionally substituted with alkyl" means that the alkyl group may but need not be present, and the description includes instances where the heterocyclyl group is substituted with alkyl group and instances where the heterocyclyl group is not substituted with alkyl group. DETAILED DESCRIPTION

[0025] The following embodiments illustrate the technical solutions of the present invention in detail, but the protection scope of the present invention includes but is not limited to them.

[0026] The structures of the compounds were determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). NMR shifts (δ) were measured in 10 -6 The unit of (ppm) is given. NMR measurements were performed using Bruker Avance III 400 and Bruker Avance 300 NMR spectrometers, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD) as the measuring solvent, and tetramethylsilane (TMS) as the internal standard;

[0027] MS was determined using Agilent 6120B (ESI) and Agilent 6120B (APCI);

[0028] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) uses a specification of 0.15mm-0.20mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm-0.5mm;

[0029] Column chromatography generally uses Yantai Huanghai Silica Gel 200-300 mesh silica gel as the carrier. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 : Twitch duration and side-lying time in rats.

[0031] Note: Compared with group G1, *** indicates P ≤ 0.001.

[0032] Example 1

[0033] (R)-1-(2-chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3carbamate and (S)-1-(2-chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3carbamate Compounds 1-1 and 1-2

[0034] (R)-1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3 carbamateand(S)-1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3 carbamate

[0035]

[0036] first step

[0037] 1-(2-Chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethan-1-one 1b

[0038] 1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethan-1-one

[0039] In a 1L three-necked flask, add 1a (100g, 429mmol) and acetonitrile (400mL), slowly add tetrazole (36g, 514.8mmol), and finally add potassium carbonate (72g, 514.8mmol). After the addition is completed, heat to 45℃ for 2h, monitor the reaction completion by TLC, and cool to room temperature. Filter to remove potassium carbonate, then add ethyl acetate (300mL×3) for extraction, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE:EA=3:1, v / v) to obtain 1b (32g, yield 33.6%, light yellow solid).

[0040] 1 H NMR (400MHz, DMSO-d6) δ9.11(s,1H),8.10(m,1H),7.67-7.63(m,2H),7.61-7.54(m,1H),6.58(m,2H).

[0041] Step 2

[0042] 1-(2-Chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethane-1,2,2-d3-1-ol 1c

[0043] 1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethan-1,2,2-d3-1-ol

[0044] Add 1b (1g, 4.48mmol) to a 100mL three-necked flask, dissolve it with 10mL deuterated methanol, add potassium carbonate (1.25g, 8.96mmol), and stir for 10min. Slowly add sodium borodeuteride (564.48mg, 13.44mmol) under ice bath, and react for 2h. LC-MS monitors the complete conversion of raw materials, slowly pour the reaction solution into water (30mL), precipitate solid, wash the solid with water three times (20mL×3) to obtain intermediate 1c, white solid (720mg, yield 69%).

[0045] 1 H NMR (400MHz, DMSO-d6) δ8.97(s,1H),7.65(dd,1H),7.46(dd,1H),7.41(td,1H),7.36(td,1H),6.02(s,1H).

[0046] Step 3

[0047] 1-(2-Chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3 carbamate compound 1

[0048] 1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3 carbamate

[0049] Add 1c (720 mg, 3.2 mmol) and tetrahydrofuran (15 mL) to a 100 mL three-necked flask, cool to -10 ° C, and slowly add sulfonyl chloride isocyanate (543.47 mg, 3.84 mmol). After the addition is completed, continue the reaction for 1 h. TLC monitors the complete conversion of the raw materials, add 10 mL of water, and stir at room temperature for 2 h. Add ethyl acetate (20 mL × 3) and extract with water, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE: EA (v / v) = 2: 1) to obtain compound 1 as a white solid (735 mg, yield 85%).

[0050] 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.54–7.50(m,1H),7.49–7.44(m,1H),7.44–7.41(m,1H),7.41–7.38(m,1H),6.91(s,1H),6.63(s,1H).

[0051] Step 4

[0052] (R)-1-(2-chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3carbamate and (S)-1-(2-chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3carbamate Compounds 1-1 and 1-2

[0053] (R)-1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3 carbamateand(S)-1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3 carbamate

[0054] Compound 1 was separated by SFC to obtain compound 1-1 (273 mg, yield 37.9%, RT = 8.188 min, 99.62% ee) and compound 1-2 (284 mg, yield 39.4%, RT = 7.155 min, 99.18% ee). Chiral HPLC (AS) mobile phase: n-hexane / ethanol = 90 / 10; column temperature: 35 ° C; column pressure: 80 bar; flow rate: 1 mL / min; detector signal channel: 215 nm @ 4.8 nm; diode array detector start and end wavelength: 200 ~ 400 nm.

[0055] Compound 1-1: 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.57–7.52(m,1H),7.48–7.45(m,1H),7.44–7.41(m,1H),7.41–7.38(m,1H),6.94(s,1H),6.66(s,1H).

[0056] Compound 1-2: 1 H NMR (400MHz, DMSO-d6) δ8.96(s,1H),7.55–7.50(m,1H),7.49–7.43(m,1H),7.42–7.37(m,1H),7.36–7.33(m,1H),6.99(s,1H),6.62(s,1H).

[0057] Example 2

[0058] (R)-1-(2-chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate and (S)-1-(2-chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate Compounds 2-1 and 2-2

[0059] (R)-1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate and(S)-1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate

[0060]

[0061] first step

[0062] 1-(2-Chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethane-2,2-d2-1-ol 2b

[0063] 1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethan-2,2-d2-1-ol

[0064] 1b (1.0 g, 4.48 mmol) was added to a 100 mL three-necked flask and dissolved in 10 mL of deuterated methanol. Potassium carbonate (1.25 g, 8.96 mmol) was added, sodium borodeuteride (564.48 mg, 13.44 mmol) was slowly added under ice bath cooling, and the reaction was continued for 2 h. LC-MS monitored that the raw material conversion was complete, and the reaction solution was slowly poured into water (30 mL) to precipitate a solid, which was washed with water three times (20 mL × 3) to obtain intermediate 2b as a white solid (680 mg, yield 69%).

[0065] 1 H NMR (400MHz, DMSO-d6) δ8.97(s,1H),7.64(dd,1H),7.46(dd,1H),7.41(td,1H),7.36(td,1H),6.03(s,1H),5.47(s,1H).

[0066] Step 2

[0067] 1-(2-Chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate compound 2

[0068] 1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate

[0069] Add 2b (720 mg, 3.2 mmol) and 15 mL of tetrahydrofuran to a 100 mL three-necked flask, cool to -10 °C, slowly add sulfonyl chloride isocyanate (543.47 mg, 3.84 mmol), and react for 1 h. TLC shows that the raw material is completely converted. Add 10 mL of water, stir at room temperature for 2 h, add ethyl acetate (20 mL × 3) to extract, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE: EA (v / v) = 2: 1) to obtain compound 2 as a white solid (680 mg, yield 85%).

[0070] 1 H NMR(400MHz,DMSO-d6)δ8.99(s,1H),7.55–7.50(m,1H),7.49–7.44(m,1H),7 .44–7.41(m,1H),7.41–7.38(m,1H),6.91(s,1H),6.65(s,1H),6.37(s,1H).

[0071] Step 3

[0072] (R)-1-(2-chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate and (S)-1-(2-chlorophenyl)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate Compounds 2-1 and 2-2

[0073] (R)-1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate and(S)-1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate

[0074] Compound 2 was separated by SFC to obtain compound 2-1 (258 mg, yield 37.9%, RT = 19.363 min, 99.62% ee) and compound 2-2 (265 mg, yield 38.9%, RT = 16.848 min, 100% ee). Chiral HPLC (AS) mobile phase: n-hexane / ethanol = 95 / 5; column temperature: 35 ° C; column pressure: 80 bar; flow rate: 1 mL / min; detector signal channel: 215 nm @ 4.8 nm; diode array detector start and end wavelength: 200 ~ 400 nm.

[0075] Compound 2-1: 1 H NMR(400MHz,DMSO-d6)δ8.99(s,1H),7.56–7.52(m,1H),7.48–7.44(m,1H),7 .44–7.42(m,1H),7.42–7.38(m,1H),6.92(s,1H),6.63(s,1H),6.37(s,1H).

[0076] Compound 2-2: 1 H NMR(400MHz,DMSO-d6)δ8.99(s,1H),7.55–7.52(m,1H),7.48–7.44(m,1H),7 .44–7.42(m,1H),7.42–7.38(m,1H),6.91(s,1H),6.63(s,1H),6.36(s,1H).

[0077] Example 3

[0078] (R)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3carbamate and (S)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3carbamate Compounds 3-1 and 3-2

[0079] (R)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate and(S)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate

[0080]

[0081]

[0082] first step

[0083] 2-Bromo-1-(4-bromo-2-chlorophenyl)ethan-1-one 3b

[0084] 2-bromo-1-(4-bromo-2-chlorophenyl)ethan-1-one

[0085] In a 250mL three-necked flask, 3a (10g, 42.9mmol) and acetonitrile (100mL) were added to dissolve, p-toluenesulfonic acid monohydrate (12.3g, 64.3mmol) was added, and succinimide (9.16g, 51.5mmol) was slowly added. After the addition was completed, the mixture was moved to 80°C and refluxed for 2h. The conversion of the raw materials was monitored by UPLC. The reaction was quenched with potassium bisulfite, and ethyl acetate (200mL×3) was added to extract with water, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The crude product was purified by column chromatography (PE:EA (v / v) = 10:1) to obtain intermediate 3b, a light yellow oil (12.5g, yield 93.9%).

[0086] Step 2

[0087] 1-(4-Bromo-2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethan-1-one 3c

[0088] 1-(4-bromo-2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethan-1-one

[0089] In a 250mL three-necked flask, add 3b (12.5g, 40.3mmol) and acetonitrile (100mL), slowly add tetrazole (3.4g, 48.4mmol), and finally add potassium carbonate (6.68g, 48.4mmol). After the addition is completed, the temperature is raised to 45°C for 2h. The conversion of the raw materials is complete when monitored by TLC, and then cooled to room temperature. Filter to remove potassium carbonate, then add ethyl acetate (200mL×3) for extraction, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v)=3:1) to obtain 3c as a light yellow solid (5.1g, yield 33.6%).

[0090] 1 H NMR (400MHz, DMSO-d6) δ9.10(s,1H),8.30(d,1H),7.85(d,1H),7.61(d,1H),6.58(s,2H).

[0091] Step 3

[0092] 1-(2-Chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethan-1-one 3d

[0093] 1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethan-1-one

[0094] In a 100mL three-necked flask, add 3c (3g, 10mmol) and dimethyl sulfoxide (30mL), then add sodium deuterated formate (760mg, 11mmol), tri-tert-butyl phosphine (202mg, 1mmol), tris(dibenzylidene indeneacetone)dipalladium (460mg, 0.5mmol), and protect with nitrogen. After the addition is completed, the temperature is raised to 80°C for 2h. The conversion of the raw materials is complete when monitored by TLC, and the mixture is cooled to room temperature. Ethyl acetate (100mL×3) is added for extraction, dried over anhydrous sodium sulfate, filtered, and the organic solvent is removed under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v)=3:1) to obtain 3d as a light yellow solid (1.8g, yield 80%).

[0095] 1 H NMR (400MHz, DMSO-d6) δ9.10(s,1H),8.29(d,1H),7.86(d,1H),7.65(d,1H),6.58(s,2H).

[0096] Step 4

[0097] 1-(2-Chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethane-1,2,2-d3-1-ol 3e

[0098] 1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethan-1,2,2-d3-1-ol

[0099] In a 50 mL three-necked flask, add 3d (446 mg, 2 mmol) deuterated methanol (5 mL) to dissolve, add potassium carbonate (560 mg, 4 mmol), stir for 10 min, and slowly add sodium borodeuteride (252 mg, 6 mmol). After the addition is completed, react for 10 min, and LC-MS monitors the complete conversion of the raw materials. Add ethyl acetate (20 mL × 3) to extract, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE: EA (v / v) = 3: 1) to obtain intermediate 3e, a light yellow oil (430 mg, yield 96%).

[0100] LC-MS m / z(ESI)=229.08[M+1].

[0101] Step 5

[0102] 1-(2-Chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3 carbamate compound 3

[0103] 1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3 carbamate

[0104] In a 50mL three-necked flask, add 3e (420mg, 3.2mmol) and tetrahydrofuran (15mL), cool to -10°C, and slowly add sulfonyl chloride isocyanate (543.47mg, 3.84mmol). After the addition is complete, react for 1h, and the reaction is complete when monitored by TLC. Add water (10mL) for hydrolysis, stir at room temperature for 2h, then add ethyl acetate (20mL×3) for water extraction, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v) = 2:1) to obtain compound 3, a white oil (220mg, yield 47%).

[0105] 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.52(d,1H),7.47(d,1H),7.42(dd,1H),6.91(s,1H),6.63(s,1H).

[0106] Step 6

[0107] (R)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3carbamate and (S)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3carbamate Compounds 3-1 and 3-2

[0108] (R)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate and(S)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate

[0109] Compound 3 was separated by SFC to obtain compound 3-1 (95 mg, yield 43.1%, RT = 19.604 min, 100% ee) and compound 3-2 (87 mg, yield 39.5%, RT = 17.280 min, 100% ee). Chiral HPLC (AS) mobile phase: n-hexane / ethanol = 95 / 5; column temperature: 35 ° C; column pressure: 80 bar; flow rate: 1 mL / min; detector signal channel: 215 nm @ 4.8 nm; diode array detector start and end wavelength: 200 ~ 400 nm.

[0110] Compound 3-1: 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.52(d,1H),7.47(d,1H),7.42(dd,1H),6.91(s,1H),6.63(s,1H).

[0111] Compound 3-2: 1 H NMR (400MHz, DMSO-d6) δ8.97(s,1H),7.51(d,1H),7.45(d,1H),7.43(dd,1H),6.91(s,1H),6.63(s,1H).

[0112] Example 4

[0113] (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3carbamate and (S)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3carbamate Compounds 4-1 and 4-2

[0114] (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate and(S)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate

[0115]

[0116] first step

[0117] 2-Bromo-1-(5-bromo-2-chlorophenyl)ethan-1-one 4b

[0118] 2-bromo-1-(5-bromo-2-chlorophenyl)ethan-1-one

[0119] In a 250mL three-necked flask, add 4a (8g, 34.3mmol) to dissolve in acetonitrile (100mL), add p-toluenesulfonic acid monohydrate (9.78g, 51.45mmol), and slowly add succinimide (7.3g, 41.2mmol). After the addition is completed, transfer to 80℃ reflux for 2h, and UPLC monitors the complete conversion of the raw materials. The reaction is quenched with potassium bisulfite, and ethyl acetate (100mL×3) is added to extract with water, dried over anhydrous sodium sulfate, filtered, and the organic solvent is removed under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v)=10:1) to obtain intermediate 4b, a light yellow oil (9.1g, yield 85%).

[0120] Step 2

[0121] 1-(5-Bromo-2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethan-1-one 4c

[0122] 1-(5-bromo-2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethan-1-one

[0123] In a 250mL three-necked flask, add 4b (9.1g, 29.3mmol) and acetonitrile (100mL), slowly add tetrazole (2.47g, 35.2mmol), and finally add potassium carbonate (4.86g, 35.2mmol). After the addition is completed, heat to 45°C for 2h, TLC monitors the complete conversion of the raw materials, and cool to room temperature. Filter to remove potassium carbonate, then add ethyl acetate (200mL×3) for extraction, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v)=3:1) to obtain 4c as a light yellow solid (3.5g, yield 38%)

[0124] 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.55(d,1H),7.46(d,1H),7.40(d,1H),6.95(s,1H),6.66(s,1H).

[0125] Step 3

[0126] 1-(2-Chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethan-1-one 4d

[0127] 1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethan-1-one

[0128] In a 1L three-necked flask, add 4c (3.1g, 10mmol) and dimethyl sulfoxide (30mL), then add sodium deuterated formate (760mg, 11mmol), tri-tert-butyl phosphine (202mg, 1mmol), tris(dibenzylidene indeneacetone)dipalladium (460mg, 0.5mmol), and protect with nitrogen. After the addition is completed, heat to 80℃ for 2h, monitor the reaction to be complete by TLC, and cool to room temperature. Add ethyl acetate (100mL×3) for extraction, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v)=3:1) to obtain 4d as a light yellow solid (1.1g, yield 44%).

[0129] 1 H NMR (400MHz, DMSO-d6) δ9.10(s,1H),8.29(d,1H),7.86(d,1H),7.65(d,1H),6.58(s,2H).

[0130] Step 4

[0131] 1-(2-Chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethane-1,2,2-d3-1-ol 4e

[0132] 1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethan-1,2,2-d3-1-ol

[0133] In a 50 mL three-necked flask, add 4d (330 mg, 1.5 mmol) deuterated methanol (5 mL) to dissolve, add potassium carbonate (420 mg, 3 mmol), stir for 10 min, and slowly add sodium borodeuteride (252 mg, 6 mmol). After the addition is completed, react for 10 min, and LC-MS monitors the complete conversion of the raw materials. Add ethyl acetate (20 mL × 3) to extract, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE: EA (v / v) = 3: 1) to obtain intermediate 4e, a light yellow oil (250 mg, yield 89%).

[0134] LC-MS m / z(ESI)=229.08[M+1].

[0135] Step 5

[0136] 1-(2-Chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3 carbamate compound 4

[0137] 1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate

[0138] In a 50mL three-necked flask, add 4e (250mg, 1.1mmol) and tetrahydrofuran (10mL), cool to -10°C, and slowly add sulfonyl chloride isocyanate (187mg, 1.32mmol). After the addition is complete, react for 1h, and the reaction is complete when monitored by TLC. Add water (10mL) for hydrolysis, stir at room temperature for 2h, then add ethyl acetate (20mL×3) for water extraction, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v) = 2:1) to obtain compound 4, a white oil (180mg, yield 62%).

[0139] 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.58(dd,1H),7.46(d,1H),7.44(dd,1H),6.91(s,1H),6.63(s,1H).

[0140] Step 6

[0141] (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate and (S)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate Compounds 4-1 and 4-2 (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate and (S)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate

[0142] Compound 4 was separated by SFC to obtain compound 4-1 (78 mg, yield 43.1%, RT = 19.604 min, 100% ee) and compound 4-2 (75 mg, yield 39.5%, RT = 17.280 min, 100% ee). Chiral HPLC (AS) mobile phase: n-hexane / ethanol = 95 / 5; column temperature: 35 ° C; column pressure: 80 bar; flow rate: 1 mL / min; detector signal channel: 215 nm @ 4.8 nm; diode array detector start and end wavelength: 200 ~ 400 nm.

[0143] Compound 4-1: 1H NMR (400MHz, DMSO-d6) δ8.99 (s, 1H), 7.52 (d, 1H), 7.46 (d, 1H), 7.41 (dd, 1H), 6.91 (s, 1H), 6.63 (s, 1H).

[0144] Compound 4-2: 1H NMR (400MHz, DMSO-d6) δ8.99 (s, 1H), 7.50 (d, 1H), 7.48 (d, 1H), 7.43 (dd, 1H), 6.92 (s, 1H), 6.63 (s, 1H).

[0145] Example 5

[0146] (R)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate and (S)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate Compounds 5-1 and 5-2

[0147] (R)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamateand(S)-1-(2-ch lorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate

[0148]

[0149] first step

[0150] 1-(2-Chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethane-2,2-d2-1-ol 5a

[0151] 1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethan-2,2-d2-1-ol

[0152] In a 50mL three-necked flask, add 3d (446mg, 2mmol), dissolve in deuterated methanol (5mL), add potassium carbonate (560mg, 4mmol), stir for 10min, and slowly add sodium borohydride (252mg, 6mmol). After the addition is completed, react for 10min, and LC-MS monitors the complete conversion of the raw materials. Add ethyl acetate (20mL×3) for extraction. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by medium pressure preparation to obtain intermediate 5a as a white solid (280mg, yield 61.5%).

[0153] LC-MS m / z(ESI)=229.08[M+1].

[0154] Step 2

[0155] 1-(2-Chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate compound 5

[0156] 1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate

[0157] In a 50mL three-necked flask, add 5a (280mg, 1.23mmol) and tetrahydrofuran (5mL) under nitrogen protection, cool to minus 10°C, and slowly add sulfonyl chloride isocyanate (208mg, 1.48mmol). After the addition is complete, react for 1h, and the reaction is complete when monitored by TLC. Add water (10mL) for hydrolysis, stir at room temperature for 2h, and then add ethyl acetate (20mL×3) for extraction. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by medium pressure preparation (water: acetonitrile = 1:1) to obtain compound 5 as a white solid (220mg, yield 47%).

[0158] 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.52(d,1H),7.46(d,1H),7.40(dd,1H),6.92(s,1H),6.63(s,1H),6.37(s,1H).

[0159] Step 3

[0160] (R)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate and (S)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate Compounds 5-1 and 5-2

[0161] (R)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamateand(S)-1-(2-chlorophenyl-4-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate

[0162] Compound 5 was separated by SFC to obtain compound 5-1 (108 mg, yield 48.1%, RT = 19.256 min, 99.52% ee) and compound 5-2 (104 mg, yield 47.4%, RT = 16.653 min, 100% ee). Chiral HPLC (AS) mobile phase: n-hexane / ethanol = 95 / 5; column temperature: 35 ° C; column pressure: 80 bar; flow rate: 1 mL / min; detector signal channel: 215 nm @ 4.8 nm; diode array detector start and end wavelength: 200-400 nm.

[0163] Compound 5-1: 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.52(d,1H),7.47(d,1H),7.42(dd,1H),6.92(s,1H),6.63(s,1H),6.37(s,1H).

[0164] Compound 5-2: 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.52(d,1H),7.47(d,1H),7.42(dd,1H),6.92(s,1H),6.63(s,1H),6.36(s,1H).

[0165] Example 6

[0166] (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate and (S)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate Compound 6-1 and Compound 6-2

[0167] (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamateand(S)-1-(2-ch lorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate

[0168]

[0169] first step

[0170] 1-(2-Chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethane-2,2-d2-1-ol 6a

[0171] 1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethan-2,2-d2-1-ol

[0172] In a 50mL three-necked flask, add 4d (446mg, 2mmol), dissolve in deuterated methanol (5mL), add potassium carbonate (560mg, 4mmol), stir for 10min, and slowly add sodium borohydride (252mg, 6mmol). After the addition is completed, react for 10min, and LC-MS monitors the complete conversion of the raw materials. Add water (10mL) to quench, and add ethyl acetate (20mL×3) to extract. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by medium pressure preparation to obtain intermediate 6a as a white solid (280mg, yield 61.5%).

[0173] LC-MS m / z(ESI)=229.08[M+1].

[0174] Step 2

[0175] 1-(2-Chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate compound 6

[0176] 1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate

[0177] In a 50mL three-necked flask, add 6a (280mg, 1.23mmol) and tetrahydrofuran (5mL) under nitrogen protection, cool to -10°C, and slowly add sulfonyl chloride isocyanate (208mg, 1.48mmol). After the addition is complete, react for 1h, and the reaction is complete when monitored by TLC. Add water (10mL) for hydrolysis, stir at room temperature for 2h, and then add ethyl acetate (20mL×3) for extraction. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is prepared by medium pressure (water: acetonitrile = 1:1) and purified to obtain compound 6 as a white solid (220mg, yield 47%).

[0178] 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.52(d,1H),7.46(d,1H),7.40(dd,1H),6.92(s,1H),6.63(s,1H),6.37(s,1H).

[0179] Step 3

[0180] (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate and (S)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethyl-2,2-d2carbamate Compound 6-1 and Compound 6-2

[0181] (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamateand(S)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate

[0182] Compound 6 was separated by SFC to obtain compound 6-1 (84.5 mg, yield 42.3%, RT = 19.213 min, 99.88% ee) and compound 6-2 (86.2 mg, yield 43.1%, RT = 16.739 min, 100% ee). Chiral HPLC (AS) mobile phase: n-hexane / ethanol = 95 / 5; column temperature: 35 ° C; column pressure: 80 bar; flow rate: 1 mL / min; detector signal channel: 215 nm @ 4.8 nm; diode array detector start and end wavelength: 200 ~ 400 nm.

[0183] Compound 6-1: 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.52(d,1H),7.46(d,1H),7.40(dd,1H),6.92(s,1H),6.63(s,1H),6.37(s,1H).

[0184] Compound 6-2: 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.52(d,1H),7.46(d,1H),7.40(dd,1H),6.92(s,1H),6.63(s,1H),6.37(s,1H).

[0185] Example 7

[0186] (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2carbamate and (S)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate

[0187] (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamateand(S)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate

[0188]

[0189]

[0190] first step

[0191] 1-(2-Chlorophenyl-5-d)-2-(2H-tetrazolyl-2-yl)ethane-2,2-d2-1-ol

[0192] 1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethan-2,2-d2-1-ol

[0193] In a 50mL three-necked flask, add 5d (446mg, 2mmol), dissolve in deuterated methanol (5mL), add potassium carbonate (560mg, 4mmol), stir for 10 minutes, and slowly add sodium borohydride (252mg, 6mmol). After the addition is completed, react for 10 minutes, and LC-Ms monitors the complete conversion of the raw materials. Add water (10mL) to quench, and add ethyl acetate (20mL×3) to extract. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by medium pressure preparation to obtain intermediate 7a as a white solid (280mg, yield 61.5%).

[0194] LC-MS m / z(ESI)=228.08[M+1].

[0195] Step 2

[0196] (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2carbamate and (S)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate

[0197] (R)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamateand(S)-1-(2-chlorophenyl-5-d)-2-(2H-tetrazol-2-yl)ethyl-2,2-d2 carbamate

[0198] In a 50mL three-necked flask, add 7a (280mg, 1.23mmol) and tetrahydrofuran (5mL) under nitrogen protection, cool to -10℃, and slowly add sulfonyl chloride isocyanate (208mg, 1.48mmol). After the addition is completed, react for 1h, and the reaction is complete when monitored by TLC. Add water (10mL) for hydrolysis, stir at room temperature for 2h, and then add ethyl acetate (20mL×3) for extraction. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by medium pressure preparation (water: acetonitrile) = 1:1 to obtain compound 7 as a white solid (200mg, yield 43%).

[0199] 1H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.52(d,1H),7.46(d,1H),7.40(dd,1H),6.92(s,1H),6.63(s,1H),6.37(s,1H).

[0200] LC-MS m / z(ESI)=271.07[M+1].

[0201] Compound 7 was separated by SFC to obtain compound 7-1 (84.5 mg, yield 42.3%, RT = 19.213 min, 99.88% ee) and compound 7-2 (86.2 mg, yield 43.1%, RT = 16.739 min, 100% ee). Chiral HPLC (AS) mobile phase: n-hexane / ethanol = 95 / 5; column temperature: 35 ° C; column pressure: 80 bar; flow rate: 1 mL / min; detector signal channel: 215 nm @ 4.8 nm; diode array detector start and end wavelength: 200 ~ 400 nm.

[0202] Compound 7-1: 1H NMR (400MHz, DMSO-d6) δ8.99 (s, 1H), 7.52 (d, 1H), 7.46 (d, 1H), 7.40 (dd, 1H), 6.92 (s, 1H), 6.63 (s, 1H), 6.37 (s, 1H).

[0203] Compound 7-2: 1H NMR (400 MHz, DMSO-d6) δ 8.99 (s, 1H), 7.52 (d, 1H), 7.46 (d, 1H), 7.40 (dd, 1H), 6.92 (s, 1H), 6.63 (s, 1H), 6.37 (s, 1H).

[0204] Example 8

[0205] (R)-1-(2-chlorophenyl-4,5-d2)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate and (S)-1-(2-chlorophenyl-4,5-d2)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate

[0206] (R)-1-(2-chlorophenyl-4,5-d2)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate and(S)-1-(2-chlorophenyl-4,5-d2)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate

[0207]

[0208] first step

[0209] 1-(5-Bromo-2-chloro-4-methoxyphenyl)ethan-1-one

[0210] 1-(5-bromo-2-chloro-4-methoxyphenyl)ethan-1-one

[0211] In a 250mL three-necked flask, add 8a (2.2g, 10mmol), dissolve in dichloromethane (20mL), and slowly add acetyl chloride (863.5mg, 11mmol) under nitrogen protection. After the addition is completed, react at 0℃ for 2h, and the plate is monitored to monitor the complete conversion of the raw materials. The reaction is quenched with saturated sodium hydroxide solution (10mL). Ethyl acetate (40mL×3) is added for extraction. The organic phases are combined, dried over anhydrous sodium sulfate, filtered, and the organic solvent is removed under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v) = 10:1) to obtain intermediate 8b as a white solid (1.8g, yield 68%).

[0212] 1H NMR (400MHz, DMSO-d6) δ8.02(s,1H),7.27(s,1H),3.94(s,3H),2.57(s,3H).

[0213] Step 2

[0214] 2-Bromo-1-(5-bromo-2-chloro-4-methoxyphenyl)ethan-1-one

[0215] 2-bromo-1-(5-bromo-2-chloro-4-methoxyphenyl)ethan-1-one

[0216] In a 100mL three-necked flask, add 8b (1.8g, 6.77mmol), dissolve in acetonitrile (20mL), add p-toluenesulfonic acid monohydrate (1.54g, 8.12mmol), and slowly add succinimide (1.45g, 8.12mmol). After the addition is completed, move to 80℃ for 2h, and UPLC monitors the complete conversion of the raw materials. The reaction is quenched with saturated potassium bisulfite aqueous solution (10mL), and ethyl acetate (30mL×3) is added for extraction. The organic phases are combined, dried over anhydrous sodium sulfate, filtered, and the organic solvent is removed under reduced pressure. The crude product is purified by silica gel column chromatography (PE:EA (v / v) = 10:1) to obtain intermediate 8c as a white solid (1g, yield 47%).

[0217] Step 3

[0218] 1-(5-Bromo-2-chloro-4-methoxyphenyl)-2-(2H-tetrazolyl-2-yl)ethan-1-one

[0219] 1-(5-bromo-2-chloro-4-methoxyphenyl)-2-(2H-tetrazol-2-yl)ethan-1-one

[0220] In a 50mL three-necked flask, add 8c (1g, 3mmol) and acetonitrile (20mL), slowly add tetrazole (252mg, 3.6mmol), and finally add potassium carbonate (496mg, 3.6mmol). After the addition is completed, heat to 45℃ for 2h, TLC monitors the complete conversion of the raw materials, and cool to room temperature. Filter to remove potassium carbonate, then add ethyl acetate (30mL×3) for extraction. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by silica gel column chromatography (PE:EA (v / v) = 3:1) to obtain 8d as a light yellow solid (300mg, yield 34%)

[0221] 1H NMR (400MHz, DMSO-d6) δ9.07(s,1H),8.38(s,1H),7.38(s,1H),6.57(s,2H),3.99(s,3H).

[0222] Step 4

[0223] 1-(5-Bromo-2-chloro-4-hydroxyphenyl)-2-(2H-tetrazolyl-2-yl)ethan-1-one

[0224] 1-(5-bromo-2-chloro-4-hydroxyphenyl)-2-(2H-tetrazol-2-yl)ethan-1-one

[0225] In a 10mL reaction bottle, add 8d (300mg, 1mmol) and N,N-dimethylformamide (5mL), and then add trimethylsilyl iodide (2g, 10mmol). After the addition is completed, heat to 120℃ and react for 24h. TLC monitors the complete conversion of the raw materials and cool to room temperature. Add water (10mL) to quench, add ethyl acetate (10mL×3) to extract. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v) = 3:1) to obtain 8e as a white solid (200mg, yield 80%)

[0226] Step 5

[0227] 4-(2-(2H-tetrazol-2-yl)acetyl)-2-bromo-5-chlorophenyl trifluoromethanesulfonate

[0228] 4-(2-(2H-tetrazol-2-yl)acetyl)-2-bromo-5-chlorophenyltrifluoromethanesulfonate

[0229] In a 50mL three-necked flask, add 8e (200mg, 0.63mmol) and tetrahydrofuran (10mL) under nitrogen protection, cool to 0℃, slowly add pyridine (99.54mg, 1.26mmol), and finally add trifluoromethanesulfonic anhydride (270.7mg, 0.96mmol). After the addition is completed, react for 1h, and the reaction is complete when monitored by TLC. Add water to quench, add ethyl acetate (20mL×3) to extract. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v) = 3:1) to obtain intermediate 8f, a white oil (200mg, yield 47%).

[0230] Step 6

[0231] 1-(2-Chlorophenyl-4,5-d2)-2-(2H-tetrazolyl-2-yl)ethan-1-one

[0232] 1-(2-chlorophenyl-4,5-d2)-2-(2H-tetrazol-2-yl)ethan-1-one

[0233] In a 100mL three-necked flask, add 8f (200mg, 0.45mmol) and dimethyl sulfoxide (5mL) under nitrogen protection, and then add sodium deuterated formate (61.6mg, 0.9mmol), tri-tert-butyl phosphine (18.2mg, 0.09mmol), tris(dibenzylidene indene acetone) dipalladium (41mg, 0.045mmol). After the addition is completed, the temperature is raised to 80℃ for 2h. The reaction is complete when monitored by TLC, and then cooled to room temperature. Water (80mL) is added to quench, and ethyl acetate (100mL×3) is added for extraction. The organic phases are combined, dried over anhydrous sodium sulfate, filtered, and the organic solvent is removed under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v) = 3:1) to obtain 8g, a light yellow solid (90mg, yield 73%).

[0234] Step 7

[0235] 1-(2-Chlorophenyl-4,5-d2)-2-(2H-tetrazolyl-2-yl)ethyl-1,2,2-d3-1-ol

[0236] 1-(2-chlorophenyl-4,5-d2)-2-(2H-tetrazol-2-yl)ethan-1,2,2-d3-1-ol

[0237] In a 50mL three-necked flask, add 8f (90mg, 0.41mmol) deuterated methanol (5mL) to dissolve, add potassium carbonate (115mg, 0.82mmol), stir for 10 minutes, and slowly add sodium borodeuteride (69mg, 1.64mmol). After the addition is completed, react for 10 minutes, and LC-Ms monitors the complete conversion of the raw materials. Add ethyl acetate (20mLX3) for extraction. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and remove the organic solvent under reduced pressure. The crude product is purified by column chromatography (PE:EA (v / v) = 3:1) to obtain 8g of the intermediate, a light yellow oil (70mg, yield 90%). LC-MS m / z (ESI) = 230.1 [M+1].

[0238] Step 8

[0239] (R)-1-(2-chlorophenyl-4,5-d2)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate and (S)-1-(2-chlorophenyl-4,5-d2)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate

[0240] (R)-1-(2-chlorophenyl-4,5-d2)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate and(S)-1-(2-chlorophenyl-4,5-d2)-2-(2H-tetrazol-2-yl)ethyl-1,2,2-d3carbamate

[0241] Compound 8-1 and Compound 8-2 were obtained by referring to the synthetic methods of Compound 5-1 and Compound 5-2.

[0242] Compound 8-1: 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.52(d,1H),7.47(d,1H),6.92(s,1H),6.63(s,1H),6.37(s,1H).

[0243] Compound 8-2: 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.52(d,1H),7.47(d,1H),6.92(s,1H),6.63(s,1H),6.36(s,1H).

[0244] Reference Example 1

[0245] (R)-1-(2-Chlorophenyl)-2-(2H-tetrazol-2-yl)ethylcarbamate

[0246] (R)-1-(2-chlorophenyl)-2-(2H-tetrazol-2-yl)ethyl carbamate

[0247]

[0248] The compound of Reference Example 1 was prepared by referring to the synthesis method of patent WO2010150946.

[0249] Biological Testing

[0250] Experiment on acute epilepsy induced by pentylenetetrazol in mice

[0251] 1. Purpose of the experiment

[0252] The anti-epileptic efficacy of the compounds was evaluated using the acute epilepsy model induced by pentylenetetrazol in mice.

[0253] 2. Experimental Materials

[0254] 1) Experimental animals

[0255] ICR mice, 6-8 weeks old, 18-22 g, half male and half female, supplied by Chengdu Dashuo Experimental Animal Co., Ltd., license number SCXK(Chuan)2013-182.

[0256] 2) Reagents

[0257] Pentylenetetrazol was purchased from Sigma-Aldrich with a batch number of MKBX1160V. Before use, it was prepared with physiological saline to a solution of the required concentration.

[0258] 3) Test drug

[0259] Weigh the compound accurately before use, add DMSO, solutol HS-15 and saline as solvents to each test group, vortex mix, and prepare the required concentration solution. The final concentrations of DMSO and solutol HS-15 are 5% and 10%, respectively. Intravenous administration, the administration volume is 10mL / kg.

[0260] 3. Experimental Methods

[0261] The day before the test, the animals were fasted but not watered. On the day of the test, the animals were randomly divided into a group of the compound of the present invention, a group of the compound of Reference Example 1, and a model control group to which only the above-mentioned solvent was added, with 10 animals in each group, half male and half female. Each group of animals was first intravenously administered with a test drug solution of 40 mg / kg or a solvent, and the effect of the test drug on the animal's autonomous activity ability within 5 minutes after administration was observed and recorded. Five minutes after administration, pentylenetetrazol (95 mg / kg) was intraperitoneally administered. The observation index adopted the Racine standard (Table 1), and the seizure level of the animal was recorded. The compound was considered to be effective against epilepsy if the animal had an epileptic seizure of grade 0 to III, and the effective rate was calculated. The test results are shown in Table 2.

[0262] Table 1 Racine criteria for epileptic seizures

[0263] Seizure level Performance Level 0 No response Level I Rhythmic twitching of the corners of the mouth, ears, or facial muscles Level II Head nodding or tail flicking, with more severe facial tics Level III Forelimb clonus Level IV Multiple limb clonus or rigidity V-Class Generalized tonic-clonic seizures, even falls

[0264] Table 2 Antiepileptic drug efficacy

[0265] Compound Efficiency Autonomous activity ability 3-1 100％ No abnormality Reference Example 1 80％ 10 hind limb weakness

[0266] Conclusion: The compound of the present invention has significant anti-epileptic effect on the acute epilepsy model of mice induced by pentylenetetrazol, and has no effect on the ability of autonomous activities.

[0267] In vivo pharmacokinetic study in mice

[0268] 1. Experimental Animals:

[0269] Species: ICR mice, SPF. Source: Chengdu Dashuo Experimental Animal Co., Ltd. Number of animals required for the experiment: 18 male mice. Animal selection: No random grouping.

[0270] 2. Experimental plan:

[0271] Eighteen ICR male mice were fasted overnight (free access to water) and divided into an intravenous (iv) administration group (9 mice) and an intragastric (ig) administration group (9 mice). 0.1 mL of blood was collected from the jugular venous plexus at 0 min, 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h after administration (3 animals at each time point). The plasma was separated by centrifugation at 4 ° C for 5 min after anticoagulation with EDTA-K2 and stored at -80 ° C for testing. The concentration of the original drug in the plasma was determined by LC / MS / MS.

[0272] 3. Drug configuration:

[0273] Weigh the drug to be tested into a 5 mL centrifuge tube, add 5% dimethyl sulfoxide and 95% 30% HP-β-CD in sequence, and after addition, mix by ultrasonication and vortexing to obtain a transparent clear solution (iv administration concentration: 0.1 mg / mL, ig administration concentration: 1 mg / mL).

[0274] 4. Preparation of standard curve:

[0275] Take 25 μL of blank mouse plasma, add 25 μL of the prepared standard solution, 25 μL of internal standard, and 225 μL of methanol, vortex mix for 2 min, centrifuge at 3200 rpm at 4°C for 20 min, and take the supernatant for LC-MS / MS analysis.

[0276] 5. Blood sample collection:

[0277] Blood was collected from the submandibular vein or other suitable veins according to the blood collection time point. About 0.03 mL of blood was collected for each sample. The samples were anticoagulated with sodium heparin and placed on ice for future use.

[0278] 6. Plasma sample processing:

[0279] Blood samples must be centrifuged to separate plasma within 1 hour (centrifugation conditions: 6800g, 6min, 2-8℃). Plasma samples must be stored in a -80℃ refrigerator before analysis.

[0280] 7. Analysis of plasma samples:

[0281] Take 25 μL of mouse plasma, 25 μL of internal standard, and 250 μL of methanol, vortex mix for 2 min, centrifuge at 3200 rpm at 4°C for 20 min, and take the supernatant for LC-MS / MS analysis.

[0282] 8. Data processing:

[0283] Detect the blood drug concentration of the test substance and draw the plasma drug concentration-time curve. When drawing the drug-time curve and calculating the parameters (may appear in C max Before and after the peak drug concentration (before and after the peak drug concentration (before and after the peak drug concentration (C )), BLQ (including “No peak”) was calculated as 0. The pharmacokinetic parameters were calculated using Phoenix WinNonlin 7.0 based on the blood drug concentration data at different time points, such as peak drug concentration (C max ), area under the curve (AUC 0-t ), half-life (T 1 / 2 ), oral bioavailability (F%), apparent clearance (CL) and steady-state distribution volume (Vss), etc. As shown in Table 3, the patented compound of the present invention has better pharmacokinetic characteristics than the reference compound.

[0284] Table 3 Pharmacokinetic parameters in mice

[0285]

[0286] Pharmacokinetic study in rats

[0287] Healthy adult SD rats weighing 180-220g were fasted overnight (free drinking water) and divided into tail vein and gavage administration groups. In the tail vein administration group, 0.1mL of blood was collected from the orbital venous plexus 5 minutes, 15 minutes, 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours before and after administration, respectively. The plasma was separated by centrifugation at 4℃ for 5 minutes and stored at -20℃ for testing. In the gavage administration group, 0.1ml of blood was collected from the orbital venous plexus 5 minutes, 15 minutes, 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours before and after administration, and the treatment method was the same as that of the intravenous injection administration group. The plasma prototype drug concentration was determined by LC-MS / MS. As shown in Table 4, the patented compounds of the present invention have better pharmacokinetic characteristics than the reference compounds.

[0288] Table 4 Pharmacokinetic parameters in rats

[0289]

[0290] Drug efficacy in rat MES model

[0291] After the SD rats were grouped, they were anesthetized with isoflurane, and cotton patches soaked in conductive liquid were attached to both ears, and the electrical stimulation wires of the experimental animals were clamped on both ears. The grouping information is detailed in Table 5. After the rats were fully awake, electrical stimulation was used to induce the rat MES model (electrical stimulation parameters: 1000mA, 50Hz, 1s). 15min before modeling, the positive compound (sodium valproate 200mg / kg) was injected into the tail vein or different doses of the compound (3mg / kg, 30mg / kg) were gavage once 15min before modeling. By observing the twitching time of the rats' limbs and tails after electrical stimulation, and the time required for the rats to stand up after the twitching ended, the results are shown in Figure 1 .

[0292] Table 5 Grouping of rats in MES experiment

[0293] Group Dosage Dosing concentration G1: Model Group iv —— G2: Sodium valproate iv 200mg / kg G3：3-2 po 30mg / kg G4：3-2 po 3mg / kg

[0294] The results show that after oral administration, the patented compound can reduce the side-lying time of rats and significantly reduce the convulsion time of rats, and has a good anti-electrical stimulation effect.

[0295] The specification of the present invention describes the specific implementation scheme in detail. Those skilled in the art should recognize that the above implementation scheme is exemplary and cannot be understood as limiting the present invention. For those skilled in the art, without departing from the principle of the present invention, by making several improvements and modifications to the present invention, the technical solutions obtained by these improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. A compound selected from:

2. A pharmaceutical composition, comprising: (1) The compound according to claim 1; (2) one or more other active ingredients; as well as (3) A pharmaceutically acceptable carrier.

3. Use of the pharmaceutical composition according to claim 2 or the compound according to claim 1 in the preparation of anti-epileptic drugs.

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