Deuterated 2-(azetidine-3-yl) ethanone compound as well as pharmaceutical composition and application thereof

Abstract

The invention discloses a deuterated 2-(azetidine-3-yl) ethanone compound as well as a pharmaceutical composition and application of the deuterated 2-(azetidine-3-yl) ethanone compound. The deuterated 2-(azetidine-3-yl) ethanone compound has the structural characteristics of a formula (I), and the definition of each group in the formula (I) is shown in the specification. The compound shown in the formula (I) can be used as a muscarinic M4 receptor positive allosteric modulator, and compared with Emraclidine (CV-231), the compound shown in the formula (I) has better pharmacokinetic characteristics and higher safety. # imgabs0 #

Description

Technical Field

[0001] This application relates to, but is not limited to, the field of pharmaceutical technology. In particular, this application relates to a deuterated 2-(azetidin-3-yl)ethanone compound, its pharmaceutical composition, and its applications. Background Art

[0002] Muscarinic acetylcholine receptors (mAChRs) are class I GPCRs (G protein-coupled receptors), consisting of 7 transmembrane segments, one extracellular N-terminus, one intracellular C-terminus, and a large intracellular segment between helix 5 and helix 6. mAChRs were first cloned and sequenced by Kubo et al. in 1986 and are encoded by the CHRM1 to CHRM5 genes, which give rise to five functionally distinct subtypes, M1 - M5. Among them, M4 is widely expressed in the striatum, caudate nucleus, and putamen and is co-expressed with dopamine receptors in striatal projection neurons, regulating dopamine release and inhibiting dopamine D1 receptor function in striatal projection neurons. M4 has become a promising target for treating the cognitive and behavioral deficits of neurodegenerative diseases and neuropsychiatric diseases.

[0003] PAMs (Positive allosteric modulations) are a class of allosteric agonists that do not directly activate receptors. By binding to allosteric sites, they increase the affinity of receptors for acetylcholine at orthosteric binding sites, thereby enhancing the response of receptors to acetylcholine. In addition, allosteric agonists do not cause receptor downregulation, probably because they do not bind to the same site as classical agonists. This advantage can effectively avoid receptor desensitization easily caused by classical agonists (Xie Kankan et al., Research progress on the expression of muscarinic receptors in schizophrenia, Neurological Diseases and Mental Health, 2021, Vol.21, No.5). Highly subtype-selective PAMs have attracted much attention because they can avoid the adverse effects caused by the activation of peripheral mAChRs. Currently, PAMs for each subtype, especially those for M4, have shown potential for anti-mental diseases and cognitive improvement in preclinical studies (Gould RW, et al., Cognitive enhancement and antipsychotic-like activity following repeated dosing with the selective M4 PAM VU0467154[J]. Neuropharmacology, 2018, 128: 492 - 502.).

[0004] Currently, there is only one M4 selective PAM in the clinical stage, namely Emraclidine (CVL-231), which is a selective positive allosteric modulator (PAM) of the M4 muscarinic receptor developed by Cerevel Therapeutics for potential oral treatment of mental disorders, including schizophrenia and Alzheimer's disease psychosis. In addition, Vanderbilt University has been working on the research and development of M4 PAMs since 2013. In the past decade, a variety of M4-targeted PAM active molecules have been introduced, such as VU0467154, VU0152099, VU0152100, etc., but they are currently in the preclinical research stage. The publicly available patent applications for selective positive allosteric modulators of the M4 muscarinic receptor include WO2018002760A1, WO2018234953A1, WO2013126856A1, WO2014035829A1, WO2017223290A1, WO2019113179A1, WO2018035444A1, WO2023064588A1, WO2023141511A1, etc.

[0005] In summary, although there is sufficient literature and drug R & D information to prove that PAMs targeting the M4 muscarinic receptor may provide new treatment approaches for the cognitive and behavioral deficits in neurodegenerative diseases and neuropsychiatric diseases, currently, no M4 muscarinic receptor PAMs have been successfully marketed. Therefore, finding compounds that are selective for the M4 muscarinic receptor and have good "drugability" is of great significance for clinical applications. Summary of the Invention

[0006] The present invention provides a new muscarinic M4 receptor positive allosteric modulator with pharmacological activity, improved pharmacokinetic and toxicological properties. The compounds provided by the present invention and their pharmaceutically acceptable salts have better pharmacokinetic characteristics and higher safety compared to Emraclidine.

[0007] In one aspect, the present invention provides a compound represented by the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, and the general formula (I) is:

[0008]

[0009] Wherein,

[0010] R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c, R 4a , R 4b , R 5a , R 5b , R 6a , R 6b , R7, R 8a , R 8b , R 9a , R 9b , R 10 , R 11 , R 12 are each independently selected from H and D, and at least one of them is selected from D.

[0011] In some embodiments, the compound of formula (I), its stereoisomers or its pharmaceutically acceptable salts, wherein R 1a , R 1b , R 1c , R2, R 3a , R 3b , R 3c , R 4a , R 4b , R 5a , R 5b , R 6a , R 6b , R7, R 8a , R 8b , R 9a , R 9b , R 10 , R 11 , R 12 1 - 21 of them are selected from D, for example, 1 - 16 are selected from D, 1 - 14 are selected from D, 1 - 12 are selected from D, 1 - 10 are selected from D, 1 - 8 are selected from D, 1 - 6 are selected from D, 1 - 4 are selected from D, 1 - 2 are selected from D, and the rest are each independently selected from H.

[0012] In some embodiments, the compound of formula (I), its stereoisomers or its pharmaceutically acceptable salts, wherein R 1a , R 1b , R 1c , R2, R 3a , R 3b , R 3c , R 4a , R 4b , R 5a , R 5b , R 6a , R 6b , R7, R 8a , R 8b , R 9a , R 9b , R 10, R 11 , R 12 Two of them are selected from D, and the rest are each independently selected from H.

[0013] In some embodiments, the compound of formula (I), its stereoisomers or its pharmaceutically acceptable salts, wherein R 1a , R 1b , R 1c , R2, R 3a , R 3b , R 3c , R 4a , R 4b , R 5a , R 5b , R 6a , R 6b , R7, R 8a , R 8b , R 9a , R 9b , R 10 , R 11 , R 12 Three of them are selected from D, and the rest are each independently selected from H.

[0014] In some embodiments, the compound of formula (I), its stereoisomers or its pharmaceutically acceptable salts, wherein R 1a , R 1b , R 1c , R2, R 3a , R 3b , R 3c , R 4a , R 4b , R 5a , R 5b , R 6a , R 6b , R7, R 8a、 R 8b , R 9a , R 9b , R 10 , R 11 , R 12 Four of them are selected from D, and the rest are each independently selected from H.

[0015] In some embodiments, the compound of formula (I), its stereoisomers or its pharmaceutically acceptable salts, wherein R 1a , R 1b , R 1c , R2, R 3a , R 3b , R 3c , R 4a , R 4b , R5a , R 5b , R 6a , R 6b , R7, R 8a , R 8b , R 9a , R 9b , R 10 , R 11 , R 12 Six of them are selected from D, and the rest are each independently selected from H.

[0016] In some embodiments, the compound represented by the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, wherein R 1a , R 1b , R 1c , R2, R 3a , R 3b , R 3c , R 4a , R 4b , R 5a , R 5b , R 6a , R 6b , R7, R 8a , R 8b , R 9a , R 9b , R 10 , R 11 , R 12 Eight of them are selected from D, and the rest are each independently selected from H.

[0017] In some embodiments, the compound represented by the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, wherein R 1a , R 1b , R 1c , R2, R 3a , R 3b , R 3c , R 4a , R 4b , R 5a , R 5b , R 6a , R 6b , R7, R 8a , R 8b , R 9a , R 9b , R 10 , R 11 , R 12 Ten of them are selected from D, and the rest are each independently selected from H.

[0018] In some embodiments, the compound of the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, wherein R 1a and R 1b and R 1c and R2 and R 3a and R 3b and R 3c and R 4a and R 4b and R 5a and R 5b and R 6a and R 6b and R7 and R 8a and R 8b and R 9a and R 9b and R 10 and R 11 and R 12 among them, 12 are selected from D, and the rest are each independently selected from H.

[0019] In some embodiments, the compound of the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, wherein R 1a and R 1b and R 1c and R2 and R 3a and R 3b and R 3c and R 4a and R 4b and R 5a and R 5b and R 6a and R 6b and R7 and R 8a and R 8b and R 9a and R 9b and R 10 and R 11 and R 12 among them, 14 are selected from D, and the rest are each independently selected from H.

[0020] In some embodiments, the compound of the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, wherein R 1a and R 1b and R 1c and R2 and R 3a and R 3b and R 3c and R 4a and R 4b and R 5a and R 5b and R 6a and R 6b and R7 and R 8a, R 8b , R 9a , R 9b , R 10 , R 11 , R 12 There are 16 selected from D, and the rest are each independently selected from H.

[0021] In some preferred embodiments of the present invention, the compound represented by the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, further the general formula (I) is as shown in the general formula (II):

[0022]

[0023] Wherein,

[0024] R 1a , R 1b , R 1c , R 3a , R 3b , R 3c , R 4a , R 4b , R 5a , R 5b , R 6a , R 6b , R 8a , R 8b , R 9a , R 9b are each independently selected from H and D, and at least 2 of them are D.

[0025] In some preferred embodiments of the present invention, the compound represented by the general formula (II), its stereoisomers or its pharmaceutically acceptable salts, wherein R 4a , R 4b , R 5a , R 5b , R 6a , R 6b , R 8a , R 8b , R 9a , R 9b at least two of them are D, and the rest are each independently selected from H.

[0026] In some preferred embodiments of the present invention, the compound represented by the general formula (II), its stereoisomers or its pharmaceutically acceptable salts, wherein R 1a , R 1b , R 1c , R 3a , R 3b , R 3c at least 3 of them are D, and the rest are each independently selected from H.

[0027] In some preferred embodiments of the present invention, the compound represented by the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, and further the general formula (I) is as shown in the general formula (III):

[0028]

[0029] Wherein,

[0030] R 1a 、R 1b 、R 1c 、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 8a 、R 8b 、R 9a 、R 9b are each independently selected from H and D;

[0031] Preferably, R 1a 、R 1b 、R 1c 、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 8a 、R 8b 、R 9a 、R 9b are each independently H.

[0032] In some preferred embodiments of the present invention, the compound represented by the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, and further the general formula (I) is as shown in the general formula (IV):

[0033]

[0034] Wherein,

[0035] R 1a 、R 1b 、R 1c 、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R6b Each independently selected from H and D.

[0036] In some preferred embodiments of the present invention, the compound represented by the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, further the general formula (I) is as shown in the general formula (V):

[0037]

[0038] Wherein,

[0039] R 1a 、R 1b 、R 1c 、R 3a 、R 3b 、R 3c 、R 6a 、R 6b 、R 8a 、R 8b 、R 9a 、R 9b Each independently selected from H and D.

[0040] In some preferred embodiments of the present invention, the compound represented by the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, further the general formula (I) is as shown in the general formula (VI):

[0041]

[0042] Wherein,

[0043] R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R 8a 、R 8b 、R 9a 、R 9b Each independently selected from H and D.

[0044] In some specific embodiments, the compound of formula (I) or its pharmaceutically acceptable salt, the compound has one of the following structures:

[0045]

[0046] On the other hand, the present invention provides a pharmaceutical composition comprising a compound of the present invention, its stereoisomer or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. In a specific embodiment, the compound of the present invention is provided in the pharmaceutical composition in an effective amount. In a specific embodiment, the compound of the present invention is provided in a therapeutically effective amount. In a specific embodiment, the compound of the present invention is provided in a prophylactically effective amount.

[0047] In some preferred embodiments of the present invention, the pharmaceutical composition is an injection, a capsule, a tablet, a pill, a powder or a granule.

[0048] On the other hand, the present invention provides an application of a compound of the present invention, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the present invention in the preparation of a medicament for the therapeutic and / or prophylactic treatment of diseases or symptoms such as Alzheimer's disease, schizophrenia or psychosis, pain, addiction, sleep disorder, cognitive disorder, depression, Parkinson's disease, Huntington's disease, movement disorder, pulmonary hypertension, chronic obstructive pulmonary disease, asthma, urinary incontinence, glaucoma, cerebral amyloid angiopathy, dementia, head trauma, stroke, pancreatitis, peripheral amyloidosis, diabetes, alcoholic liver, hepatitis and atherosclerosis.

[0049] On the other hand, the present invention provides a compound of the present invention, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the present invention for use in medicine.

[0050] On the other hand, the present invention provides a compound of the present invention, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the present invention for use in the therapeutic and / or prophylactic treatment of diseases or symptoms such as Alzheimer's disease, schizophrenia or psychosis, pain, addiction, sleep disorder, cognitive disorder, depression, Parkinson's disease, Huntington's disease, movement disorder, pulmonary hypertension, chronic obstructive pulmonary disease, asthma, urinary incontinence, glaucoma, cerebral amyloid angiopathy, dementia, head trauma, stroke, pancreatitis, peripheral amyloidosis, diabetes, alcoholic liver, hepatitis and atherosclerosis.

[0051] On the other hand, the present invention provides a method for the therapeutic and / or prophylactic treatment of diseases or symptoms such as Alzheimer's disease, schizophrenia or psychosis, pain, addiction, sleep disorder, cognitive disorder, depression, Parkinson's disease, Huntington's disease, movement disorder, pulmonary hypertension, chronic obstructive pulmonary disease, asthma, urinary incontinence, glaucoma, cerebral amyloid angiopathy, dementia, head trauma, stroke, pancreatitis, peripheral amyloidosis, diabetes, alcoholic liver, hepatitis and atherosclerosis, the method comprising administering to an individual in need thereof a therapeutically effective amount of a compound of the present invention, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the present invention.

[0052] Synthesis route

[0053] Those skilled in the art can prepare the compounds of the present invention by combining Patent WO2018002760A1 and known organic synthesis techniques. The starting materials are commercially available chemicals and / or compounds described in chemical literature. The difference lies in using deuterated starting materials instead of non-deuterated starting materials in the reaction, or obtaining them by performing a one-step deuteration reaction on known compounds.

[0054] Now, the terms used to describe the present invention in the specification and claims of the present invention are defined as follows. For specific terms, if the meaning defined in the present invention is inconsistent with the meaning commonly understood by those skilled in the art, the meaning defined in the present invention shall prevail; if it is not defined in the present invention, it shall have the meaning commonly understood by those skilled in the art.

[0055] In the present invention, the compound name and its structural formula have a corresponding relationship. When the compound name is inconsistent with the structural formula, the structural formula shall prevail, or it can be inferred according to the specific situation of the present invention in combination with the knowledge of those skilled in the art.

[0056] The term "deuterated" used in the present invention means that one or more hydrogens in a compound or group are replaced by deuterium (denoted by D in the general formula). Deuteration can be mono-substitution, di-substitution, multi-substitution or full substitution.

[0057] "Stereoisomer" refers to an isomer generated by the different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and conformational isomers.

[0058] "Pharmaceutically acceptable salt" refers to a pharmaceutically acceptable organic salt or inorganic salt of the compounds of the present invention as defined above, and the salt has the desired pharmacological activity. Such salts include acid addition salts formed with inorganic acids or organic acids. Pharmaceutically acceptable salts also include base addition salts, which can be formed in the presence of acidic protons capable of reacting with inorganic bases or organic bases.

[0059] Other features and advantages of the present application will be described in the subsequent specification, and some of them will become obvious from the specification, or be understood by implementing the present application. Other advantages of the present application can be achieved and obtained through the solutions described in the specification. Detailed implementation mode

[0060] To make the purpose, technical solution and advantages of the present application clearer, the embodiments of the present invention will be described in detail below. It should be noted that, without conflict, the embodiments in the present application and the features in the embodiments can be combined arbitrarily with each other.

[0061] The following specific examples are used to describe the implementation process of the present invention and the beneficial effects produced, which are intended to help readers better understand the essence and characteristics of the present invention, and are not intended to limit the scope of the present invention. The following examples are only used to illustrate the present application and should not be regarded as limiting the scope of the present application.

[0062] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10 -6 The units are given. NMR measurements were performed using a Bruker Avance Neo 400 MHz nuclear magnetic resonance spectrometer, the measurement solvent was deuterated dimethyl sulfoxide (DMSO-d6), and the internal standard was tetramethylsilane (TMS).

[0063] LCMS was determined using Waters ACQUITY UPLC.

[0064] High performance liquid chromatography (HPLC) was measured by a Thermo UltiMate 3000 liquid chromatograph using a Venusil ASB C18 (4.6*250 mm, 5 μm) column.

[0065] UPLC-MS / MS was determined using TQ6500+.

[0066] The thin layer chromatography silica gel plate used was West Asia Reagent GF254 silica gel plate.

[0067] Column chromatography used 200-300 mesh silica gel from Qingdao Ocean Chemical Co., Ltd. as the carrier.

[0068] The known starting materials of the present application can be synthesized by methods known in the art, or can be purchased from reagent companies such as Aladdin, Bidex Pharmaceuticals, and WuXi AppTec.

[0069] The abbreviations used herein have the following meanings:

[0070] HPLC: high performance liquid chromatography; LCMS: liquid chromatography-mass spectrometry; TLC: thin layer chromatography; 1 H NMR: hydrogen nuclear magnetic resonance; TCFH: N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate; NMI: N-methylimidazole; BINAP: 1,1′-binaphthyl-2,2′-diyl(diphenylphosphine); Pd2(dba)3: tris(dibenzylideneacetone)dipalladium(0); MeOD: deuterated methanol; DMSO: dimethyl sulfoxide; PEG400: polyethylene glycol 400; Tween-80: Tween-80.

[0071] Example 1

[0072] 1-(2,4-Dimethyl-5,7-dihydro-6H-pyrrolo[3,4-b]pyridin-6-yl)-2-(1-(2-(trifluoromethyl)pyridin-4-yl)azetidin-3-yl)ethyl-1-one-2,2-D2 (Compound 1)

[0073]

[0074] Step 1: Trifluoroacetic acid (2 mL) was added dropwise to a solution of intermediate 1_1 (500 mg, 2.2 mmol) in dichloromethane (5 mL), and the reaction mixture was stirred at room temperature for 4 hours. The solvent was removed by rotary evaporation under reduced pressure to obtain a colorless transparent oily intermediate 1_2 (271 mg, yield: 96%). ESI-MS m / z: 130.1 [M+H] + 。

[0075] Step 2: Intermediate 1_2 (2.1 mmol, 1.0 eq, 271 mg), intermediate 1_3 (5.2 mmol, 2.5 eq, 592 mg), 1,1′-Binaphthalene-2,2'-diyl(diphenylphosphine) (BINAP, 0.83 mmol, 0.4 eq, 288 mg) were added to a reaction flask, dissolved and stirred in toluene (10 mL), then cesium carbonate (10.4 mmol, 5.0 eq, 1.85 g) was added. The mixture was heated in an oil bath at 70 °C. After the temperature rose, tris(dibenzylideneacetone)dipalladium(0) (Pd2(dba)3, 0.4 mmol, 0.2 eq, 205 mg) was added, and the reaction was heated at 70 °C for 16 hours. The reaction was monitored by TLC until completion. Most of the toluene was removed by vacuum concentration, and the residue was diluted with water (3 x 20 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with saturated aqueous sodium chloride solution (2 x 10 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. Column chromatography (SiO2, eluent: cyclohexane / ethyl acetate, gradient: 10:1 to 1:1) gave a colorless transparent oily intermediate 1_4 (518 mg, yield: 90%). ESI-MS m / z: 275.1 [M+H] + 。

[0076] Step 3: Intermediate 1_4 (1.0 eq, 518 mg) was taken, dissolved and stirred in methanol (4.0 mL). The pH of the solution was adjusted to 13 with 1 M sodium hydroxide solution, and the mixture was stirred at room temperature for 10 h. The reaction was monitored by TLC until completion. 1 M dilute hydrochloric acid was added to the reaction solution to adjust the pH to 5, and the solution was concentrated under reduced pressure until completely evaporated. Then the concentrate was placed in a vacuum drying oven for further drying to obtain a white solid powder intermediate 1_5 (127 mg, yield: 26%). ESI-MS m / z: 261.1 [M+H] + 。

[0077] Weigh intermediate 1_5 (0.4 mmol, 1.0 eq, 100 mg), add analytical grade acetonitrile (2 mL), stir to dissolve, add N-methylimidazole (NMI, 1.4 mmol, 3.5 eq, 165 mg) and stir to dissolve for 15 min. Weigh intermediate 16 (0.4 mmol, 1.0 eq, 85 mg), add and stir to dissolve. Add N,N,N′,N′-tetramethylchloroformamidinium hexafluorophosphate (TCFH, 0.45 mmol, 1.1 eq, 175 mg), stir to dissolve, and stir at room temperature for 4 h. Drop the reaction solution into purified water (10 mL) for crystallization, let it stand for 10 min and then perform suction filtration. Then wash the filter cake thoroughly with purified water (10 mL, in three portions), dry at 40 °C to obtain yellow solid intermediate 1_7 (100 mg, yield 67%). ESI-MS m / z: 391.2 [M+H] + 。 1 1H NMR (600 MHz, DMSO-d6) δ 8.21 (d, J = 5.7 Hz, 1H), 7.00 (d, J = 4.5 Hz, 1H), 6.71 (d, J = 2.0 Hz, 1H), 6.53 (dd, J = 5.7, 2.2 Hz, 1H), 4.79 (d, J = 27.7 Hz, 2H), 4.77 (s, 1H), 4.77 (s, 1H), 4.57 (d, J = 17.9 Hz, 1H), 4.54 (s, 1H), 4.21 - 4.13 (m, 2H), 3.74 - 3.66 (m, 2H), 3.17 - 3.05 (m, 1H), 2.85 (d, J = 7.6 Hz, 2H), 2.42 (d, J = 1.8 Hz, 3H), 2.23 (d, J = 7.2 Hz, 3H).

[0078] Weigh intermediate 1_7 (0.4 mmol, 1.0 eq, 100 mg), add deuterated methanol (4 mL), stir to dissolve, add cesium carbonate (1.2 mmol, 3.0 eq, 391 mg), and stir at room temperature for 14 h. Concentrate under reduced pressure, purify by column chromatography (SiO2, eluent is cyclohexane / ethyl acetate, gradient is 10:1 to 1:1), and dry to obtain white solid compound 1 (79 mg, yield: 79%). ESI-MS m / z: 393.2 [M+H] + 。 11H NMR (600 MHz, DMSO-d6) δ 8.21 (d, J = 5.7 Hz, 1H), 7.01 (d, J = 5.9 Hz, 1H), 6.72 (d, J = 1.5 Hz, 1H), 6.53 (dd, J = 5.7, 2.1 Hz, 1H), 4.82 (s, 1H), 4.77 (s, 1H), 4.58 (s, 1H), 4.54 (s, 1H), 4.21 - 4.13 (m, 2H), 3.75 - 3.67 (m, 2H), 3.21 - 3.00 (m, 1H), 2.42 (d, J = 2.5 Hz, 3H), 2.23 (d, J = 7.0 Hz, 3H).

[0079] Example 2

[0080]

[0081] Synthesis of Intermediate 2_2: Take 4-bromo-2-(trifluoromethyl)pyridine (1.38 g, 6.11 mmol), Intermediate 2_1 (1 g, 9.12 mmol), cesium fluoride (927 mg, 6.11 mmol), and triethylamine (2.40 g, 23.72 mmol) in a 50 mL round-bottom flask. Add 10 mL of DMSO and stir. Heat to 90 °C under nitrogen protection and react for 12 h. Monitor the reaction by LCMS until it is complete. Cool down, add 50 mL of water to the reaction solution and stir. A large amount of white solid precipitates. Filter by suction, and dry the filter cake in a blast drying oven to obtain 1.01 g of white solid with a yield of 75.88%. ESI-MS m / z: 219.10 [M + H] + .

[0082] Synthesis of Intermediate 2_3: Take Intermediate 2_2 (1 g, 4.59 mmol) and Dess-Martin oxidant (3.9 g, 9.20 mmol) in a 50 mL round-bottom flask. Add 12 mL of dichloromethane and stir evenly. React at room temperature for 10 h. Monitor the reaction by LCMS until it is complete. After the reaction is completed, add 15 mL of dichloromethane and 50 mL of saturated sodium thiosulfate solution to the reaction solution and stir for 30 min. Let it stand and separate the layers. Collect the organic layer, add 50 mL of saturated sodium carbonate and stir for 15 min. Separate the layers. After the organic layer is evaporated to dryness, purify it by column chromatography (pure DCM) to obtain 700 mg of white powder with a yield of 70.71%. ESI-MS m / z: 249.17 [M + CH3OH + H] + .

[0083] Synthesis of Intermediate 2-4: Take Intermediate 2-3 (700 mg, 3.24 mmol), potassium tert-butoxide (1.45 g, 12.96 mmol), and 5 mL of deuterated methanol in a 50 mL round-bottom flask. React at room temperature for 18 h, and monitor the completion of the reaction by LCMS. Adjust the pH of the reaction solution to 7 by dropwise addition of glacial acetic acid, concentrate under vacuum, and purify by column chromatography to obtain 566 mg of a white solid with a yield of 79.41%. ESI-MS m / z: 239.16 [M + H2O + H] + .

[0084] Synthesis of Intermediate 2-5: Take Intermediate 2-4 (566 mg, 2.57 mmol) and ethoxycarbonylmethylenetriphenylphosphine (896 mg, 2.57 mmol) in a 50 mL round-bottom flask. Add 6 mL of dichloromethane and stir. React at room temperature for 17 h, and monitor the completion of the reaction by LCMS. Concentrate the reaction solution under vacuum. Add water (40 mL × 3) and 50 mL of dichloromethane to the residue for extraction. Collect the organic phase, dry by evaporation, and purify by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 100:1) to obtain 456 mg of a white solid with a yield of 61.21%. ESI-MS m / z: 291.16 [M + H] + .

[0085] Synthesis of Intermediate 2-6: Take Intermediate 2-5 (456 mg, 1.57 mmol) and 160 mg of Pd / C in a 50 mL round-bottom flask. Add 6 mL of dichloromethane and stir. React at room temperature for 19 h under hydrogen protection, and monitor the completion of the reaction by LC-MS. Filter the reaction solution, and dry the filtrate by evaporation to obtain 176 mg of a white powder with a yield of 38.22%. LCMS (ESI) m / z = 293.16 [M + H] + .

[0086] Synthesis of Intermediate 2-7: Take Intermediate 2-6 (176 mg, 0.60 mmol) and 6 mL of methanol in a round-bottom flask. Slowly add 1 mol / L NaOH dropwise to adjust the pH of the solution to 13 - 14. React at room temperature for 3 h, and monitor the completion of the reaction by LCMS. Concentrate the reaction solution under vacuum. Add 60 mL of water and dichloromethane (60 mL × 3) to the residue for extraction. Discard the organic phase. Adjust the pH of the aqueous phase to around 5 with 1 mol / L dilute hydrochloric acid, and lyophilize to obtain 600 mg of an intermediate containing sodium chloride. ESI-MS m / z: 265.13 [M + H] + .

[0087] Synthesis of Compound 2: Take intermediate 2_7 (105 mg, 0.40 mmol), intermediate 1_6 (88 mg, 0.40 mmol), N-methylimidazole (114 mg, 1.39 mmol, NMI), 3 mL of acetonitrile and stir evenly in a round-bottom flask. Finally, weigh N,N,N′,N′-tetramethylchloroformamidinium hexafluorophosphate (123 mg, 0.44 mmol, TCFH) and add it to the reaction solution, then react at room temperature for 3 h. Monitor the reaction by LCMS until it is complete. Concentrate the reaction solution under reduced pressure. Add 50 mL of water and dichloromethane (50 mL×3) to the residue for extraction. Combine the organic phases, dry over anhydrous sodium sulfate, and evaporate to dryness. Purify by thin-layer chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 32 mg of the final product, a white powder, with a yield of 20.43%. ESI-MS m / z: 197.82 [M / 2+1]. 1 H NMR (400 MHz, DMSO-d6) δ 8.20 (d, J = 5.7 Hz, 1H), 7.06 - 6.96 (m, 1H), 6.71 (d, J = 2.3 Hz, 1H), 6.53 (dd, J = 5.7, 2.3 Hz, 1H), 4.79 (d, J = 18.9 Hz, 2H), 4.56 (d, J = 17.5 Hz, 2H), 3.18 - 3.08 (m, 1H), 2.98 - 2.75 (m, 2H), 2.42 (s, 3H), 2.23 (d, J = 4.8 Hz, 3H).

[0088] Biological Activity Test

[0089] Biological Test Example 1: Test on the Allosteric Modulatory Activity of the Compounds of the Present Invention on Acetylcholine-Activated M4 Receptors

[0090] Experimental Purpose: Apply the FLIPR detection technology to detect the allosteric modulation of the test compound on the intracellular calcium ion release effect induced by acetylcholine-activated M4 receptors at the cellular level.

[0091] Main Experimental Materials and Sources:

[0092] DMEM (culture medium) - gibco-11965-092

[0093] Fluo-4Direct TM Kit - Invitrogen-F10471

[0094] 384-well polylysine-coated cell plate - Greiner-781090

[0095] Vi-cell XR Cell Viability Analyzer - Beckman Coulter

[0096] Compound Preparation ECHO (Acoustic Liquid Handling System) - Greiner - 781280

[0097] Experimental Protocol:

[0098] 1. Cell Preparation: Take Chinese hamster ovary cells (CHO) in logarithmic growth phase stably expressing M4 muscarinic acetylcholine receptor (M4mAChR), wash them with DPBS (Dulbecco's Phosphate Buffered Saline) buffer, add an appropriate amount of 3 mL of EDTA (Ethylenediaminetetraacetic acid) - trypsin, place them in an incubator at 37°C and 5% carbon dioxide for digestion, take out the cells after 1 - 2 minutes and add medium to terminate digestion. Disperse the cells by repeated pipetting and then centrifuge to collect the cells. Suspend the cells at a concentration of 10x10 5 cells / mL in growth medium; add 20 μL of cell suspension (20K / well) to each well of a 384 - well plate; place the cells in an incubator at 37°C and 5% CO2 overnight.

[0099] 2. FLIPR Experiment Preparation: Prepare probenecid in FLIPR assay buffer; prepare 2X (8 μM) Fluo - 4 Direct TM loading buffer (per 10 mL). Dilute it in 10 - fold steps, and then use the ECHO liquid handling system to transfer 900 nL of the compound into a 384 - well compound plate.

[0100] 3. Add Fluo - 4: Take out the cell plate from the incubator, gently remove the medium, and add 20 μL of assay buffer and 20 μL of 2X Fluo - 4 Direct TM wash - free loading buffer to the 384 - well cell culture plate using a pipette. The final volume in the cell culture plate is 40 μL.

[0101] 4. Incubation: Incubate at 37°C and 5% CO2 for 50 minutes, and then incubate at room temperature for 10 minutes.

[0102] 5. Testing: Take out the cell plate from the incubator, place it in the FLIPR, run the FLIPR instrument software, according to the set program, add 10 μL of experimental buffer solution, and read the fluorescence signal. Then add 10 μL of agonist reference compound (acetylcholine), read the fluorescence signal, and calculate EC 20 and prepare an agonist at 6×EC 20 concentration. Gradient - dilute the test compound (the compound of the embodiment of the present invention) 10 - fold in 3 steps using ECHO and transfer 900 nL to the compound plate. Add 40 μL of FLIPR buffer solution to the compound plate. Run the FLIPR instrument software, according to the set program, add 10 μL of the test compound and the reference compound to the cell plate, and read the fluorescence signal. Then add 10 μL of 6×EC 20Agonist at a concentration was added to the cell plate, and the fluorescence signal was read.

[0103] 6. Experimental evaluation method and result analysis: In the study, the functional drug screening system FLIPR was used to detect the change in intracellular calcium ion concentration. After fitting the data, the concentration value corresponding to 50% activation of the maximum calcium current (EC 50 ) and %Activity@Max Dose were calculated based on the dose-response curve to determine the in vitro cell activity of the compounds of the present invention. The experimental results were processed using the statistical software GraphPad 5.0. Fitting was performed according to the obtained data to obtain the dose-response curve and EC 50 , and the results are shown in Table 1 below

[0104] Table 1. Allosteric regulation activity results of the compounds of the present invention on acetylcholine-activated M4 receptor

[0105] Test substance <![CDATA[EC 50 (nM)]]> % Activity at maximum dose CVL-231 (Intermediate 1_7) 168.0 79.77 Compound 1 152.9 72.42

[0106] Biological test example 2: Pharmacokinetic PK test of the compounds of the present invention in mice

[0107] Experimental purpose: By administering the test substance to mice by single-dose intravenous injection and gavage, the concentration of the test substance in rat plasma was measured to evaluate the pharmacokinetic characteristics and bioavailability of the test substance in mice.

[0108] Experimental procedure (1) Preparation of experimental samples:

[0109] A) Preparation process of CVL-231 intravenous injection sample:

[0110] a) Weigh 1.6 mg of CVL-231 into a 10 mL vial;

[0111] b) Add 0.4 mL of DMSO, 2.4 mL of PEG 400, and 0.4 mL of Tween-80 to the vial, vortex evenly and then sonicate until clear and transparent;

[0112] c) Add 4.8 mL of physiological saline solution to the vial and vortex evenly;

[0113] d) Filter the prepared solution through a 0.22 μm filter membrane. Set aside.

[0114] B) Preparation process of CVL-231 gavage sample:

[0115] a) Weigh 4.8 mg of CVL-231 into a 100 mL vial;

[0116] b) Add 0.8 mL of DMSO, 4.8 mL of PEG400, and 0.8 mL of Tween-80 to the vial, vortex evenly and then sonicate until clear and transparent;

[0117] c) Add 9.6 mL of saline solution to the bottle and vortex to mix well. Set aside.

[0118] (2) Drug administration process:

[0119] a) Before drug administration, mice were taken out of the cage, weighed, and placed on a fixture;

[0120] b) Calculate the dosage based on the animal's body weight;

[0121] c) After receiving the compound, shake it well and use a syringe to pipette the sample before administration to mix it evenly;

[0122] d) draw a specified volume of intravenous (iv) sample;

[0123] e) Administration was completed via gavage, and 1 ml of air was used to flush the gavage tube to ensure that the sample was completely administered to the animal.

[0124] (3) Blood collection process:

[0125] Blood was collected directly from the eye socket using a capillary tube. After collection, the blood was placed in a vacuum tube and transferred on warm ice and centrifuged. The centrifugation time was recorded on a centrifugation table. The blood samples were used for UPLC-MS / MS (TQ6500+) analysis and detection. PK parameters were analyzed using WinNonlin 6.1 non-compartmental model.

[0126] The preparation and experimental operation of the compound 1 of the present invention were carried out in parallel with the above operation. The test results are shown in Table 2 below.

[0127] Table 2. Results of mouse PK experiments on the compounds of the present invention

[0128]

[0129] As can be seen from the data in Tables 1 and 2, the in vitro activity of the compound of the present invention is comparable to that of CV1-231, while the in vivo PK data of mice show that the bioavailability of the compound of the present invention by oral administration (po) is superior to that of CV1-231. The deuterated compound provided by the present invention can improve the oral bioavailability of the control compound, thereby hopefully overcoming the defects of individual differences in clinical medication and ultimately benefiting patients.

[0130] Although the embodiments disclosed in this application are as above, the contents described are only embodiments adopted to facilitate understanding of this application and are not intended to limit this application. Any technician in the field to which this application belongs can make any modifications and changes in the form and details of implementation without departing from the spirit and scope disclosed in this application, but the scope of protection of this application shall still be based on the scope defined in the attached claims.

Claims

1. A compound of the general formula (I), its stereoisomers or its pharmaceutically acceptable salts, wherein the general formula (I) is: Wherein, R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R7、R 8a 、R 8b 、R 9a 、R 9b 、R 10 、R 11 、R 12 Each independently selected from H and D, and at least one of them is selected from D.

2. The compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein, R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R7、R 8a 、R 8b 、R 9a 、R 9b 、R 10 、R 11 、R 12 Among them, 1 to 21 are selected from D. For example, 1 to 16 are selected from D, 1 to 14 are selected from D, 1 to 12 are selected from D, 1 to 10 are selected from D, 1 to 8 are selected from D, 1 to 6 are selected from D, 1 to 4 are selected from D, 1 to 2 are selected from D, and the rest are each independently selected from H.

3. The compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 2, wherein, R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R7、R 8a 、R 8b 、R 9a 、R 9b 、R 10 、R 11 、R 12 There are two selected from D, and the rest are each independently selected from H; or R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R7、R 8a 、R 8b 、R 9a 、R 9b 、R 10 、R 11 、R 12 Three of them are selected from D, and the rest are each independently selected from H; or R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R7、R 8a 、R 8b 、R 9a 、R 9b 、R 10 、R 11 、R 12 Four of them are selected from D, and the rest are each independently selected from H; or R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R7、R 8a 、R 8b 、R 9a 、R 9b 、R 10 、R 11 、R 12 Six of them are selected from D, and the rest are each independently selected from H; or R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R7、R 8a 、R 8b 、R 9a 、R 9b 、R 10 、R 11 、R 12 Eight of them are selected from D and the rest are each independently selected from H; or R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R7、R 8a 、R 8b 、R 9a 、R 9b 、R 10 、R 11 、R 12 There are 10 selected from D, and the rest are each independently selected from H; or R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R7、R 8a 、R 8b 、R 9a 、R 9b 、R 10 、R 11 、R 12 Twelve of them are selected from D, and the rest are each independently selected from H; or R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R7、R 8a 、R 8b 、R 9a 、R 9b 、R 10 、R 11 、R 12 There are 14 selected from D, and the rest are each independently selected from H; or R 1a 、R 1b 、R 1c 、R2、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R7、R 8a 、R 8b 、R 9a 、R 9b 、R 10 、R 11 、R 12 There are 16 selected from D, and the rest are each independently selected from H.

4. The compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein, The general formula (I) is as shown in the general formula (II): Wherein, R 1a 、R 1b 、R 1c 、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R 8a 、R 8b 、R 9a 、R 9b are each independently selected from H and D, and at least two of them are D; or R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R 8a 、R 8b 、R 9a 、R 9b At least two of them are D, and the rest are each independently selected from H; or R 1a 、R 1b 、R 1c 、R 3a 、R 3b 、R 3c At least 3 of them are D, and the rest are each independently selected from H.

5. The compound according to claim 1, its stereoisomer or its pharmaceutically acceptable salt, wherein, The general formula (I) is as shown in the general formula (III): Wherein, R 1a 、R 1b 、R 1c 、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 8a 、R 8b 、R 9a 、R 9b each independently selected from H and D; or R 1a 、R 1b 、R 1c 、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 8a 、R 8b 、R 9a 、R 9b Each independently is H.

6. The compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein, The general formula (I) is as shown in the general formula (IV): Wherein, R 1a 、R 1b 、R 1c 、R 3a 、R 3b 、R 3c 、R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b Each independently selected from H and D.

7. The compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein, The general formula (I) is as shown in the general formula (V): Wherein, R 1a 、R 1b 、R 1c 、R 3a 、R 3b 、R 3c 、R 6a 、R 6b 、R 8a 、R 8b 、R 9a 、R 9b Each independently selected from H and D.

8. The compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein, The general formula (I) is as shown in the general formula (VI): Wherein, R 4a 、R 4b 、R 5a 、R 5b 、R 6a 、R 6b 、R 8a 、R 8b 、R 9a 、R 9b are each independently selected from H and D.

9. The compound, its stereoisomers or its pharmaceutically acceptable salts according to claim 1, having one of the following structures:

10. A pharmaceutical composition comprising the compound, its stereoisomers or its pharmaceutically acceptable salts according to any one of claims 1 to 9 and a pharmaceutically acceptable excipient.

11. The compound, its stereoisomers or its pharmaceutically acceptable salts according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10, in the preparation of a drug for the therapeutic and / or prophylactic treatment of diseases or symptoms such as Alzheimer's disease, schizophrenia or psychosis, pain, addiction, sleep disorder, cognitive disorder, depression, Parkinson's disease, Huntington's disease, movement disorder, pulmonary hypertension, chronic obstructive pulmonary disease, asthma, urinary incontinence, glaucoma, cerebral amyloid angiopathy, dementia, head trauma, stroke, pancreatitis, peripheral amyloidosis, diabetes, alcoholic liver, hepatitis and atherosclerosis.

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