Nitrogen-containing heterocyclic compound, pharmaceutical composition and application thereof

Abstract

The present invention discloses a nitrogen-containing heterocyclic compound, a pharmaceutical composition and application thereof. The present invention specifically discloses a compound of formula I or a pharmaceutically acceptable salt thereof, which has one or more of the following advantages: novel structure, high drug-like properties and excellent metabolic properties. #imgabs0#

Description

Technical Field

[0001] The invention relates to nitrogen-containing heterocyclic compounds, pharmaceutical compositions and applications thereof. Background Art

[0002] Muscarinic acetylcholine receptor-mediated disorders, such as major depressive disorder (MDD), bipolar disorder (BPD), and schizophrenia, continue to be a serious public health problem. Currently available antidepressants, mood stabilizers, and antipsychotics can relieve some of the symptoms of mood disorders in some patients, but they are only partially effective for a large number of patients, and many patients are refractory to existing drug treatments. Patients with major depression typically experience at least two weeks of pervasive low mood, low self-esteem, and loss of interest or pleasure in activities that are usually pleasurable. Patients with bipolar disorder typically experience periods of depression and abnormally elevated mood, each lasting from days to weeks. Patients with schizophrenia often experience active social avoidance, passive social withdrawal, emotional withdrawal, and anxiety. In some cases, supportive therapies are used to help relieve the symptoms of muscarinic acetylcholine receptor-mediated disorders; however, supportive therapies do not directly treat the disorder and are not a long-term solution to meet the medical needs of patients.

[0003] Compounds that modulate muscarinic acetylcholine receptors are useful for treating diseases associated with muscarinic acetylcholine receptor activity, such as major depressive disorder, bipolar disorder, and schizophrenia. Muscarinic acetylcholine receptors are a type of acetylcholine receptor that forms a G protein-coupled receptor complex on the cell membranes of certain neurons and other cells. U.S. Patent 10,604,519 describes certain compounds that are active at the M4 muscarinic acetylcholine receptor, but there is still a need for new compounds with good drug-like properties to treat diseases mediated by muscarinic acetylcholine receptors. Summary of the invention

[0004] The technical problem to be solved by the present invention is that the compounds active on M4 muscarinic acetylcholine receptors in the prior art have the defects of relatively simple structure, low drug-like properties and poor metabolic properties. To this end, the present invention provides a nitrogen-containing heterocyclic compound, a pharmaceutical composition and application thereof, which has one or more of the following advantages: novel structure, high drug-like properties and excellent metabolic properties.

[0005] The present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof,

[0006]

[0007] Among them, R 1 , R 2 and R 3 Independently for C 1-6 alkyl.

[0008] In certain preferred embodiments of the present invention, certain groups of the compound of formula I or its pharmaceutically acceptable salt are defined as follows, and the unmentioned groups are the same as those described in any embodiment of the present invention (referred to as "in a certain embodiment of the present invention"). For other inventions of the present invention, the same abbreviation as the present invention is adopted and no further description is given.

[0009] In one embodiment of the present invention, R 1 , R 2 and R 3 Independently for C 1-3 An alkyl group, such as a methyl group.

[0010] In one embodiment of the present invention, R 1 It is methyl.

[0011] In one embodiment of the present invention, R 2 It is methyl.

[0012] In one embodiment of the present invention, R 3 It is methyl.

[0013] In one embodiment of the present invention, the compound of formula I is .

[0014] The present invention also provides a pharmaceutical composition, which comprises the compound of formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutical excipient.

[0015] The present invention also provides a use of the compound of formula I, a pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of a muscarinic acetylcholine receptor positive allosteric modulator.

[0016] The present invention also provides a use of the compound of formula I, a pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of a drug for treating and / or preventing mental illness;

[0017] The mental illness is major depression, bipolar disorder or schizophrenia.

[0018] The present invention also provides a use of the compound of formula I, a pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of a medicament for treating and / or preventing diseases mediated by muscarinic acetylcholine receptors.

[0019] In one embodiment of the present invention, the muscarinic acetylcholine receptor-mediated disease is an M4 muscarinic acetylcholine receptor-mediated disease.

[0020] In one embodiment of the present invention, the muscarinic acetylcholine receptor-mediated disease is Alzheimer's disease, schizophrenia, major depressive disorder or bipolar disorder.

[0021] On the basis of being in accordance with the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0022] Unless otherwise specified, the terms used in this invention may be defined as follows:

[0023] The term "alkyl" refers to a straight or branched chain alkyl group having a specified number of carbon atoms (e.g., C1-C6). Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, and the like.

[0024] The term "pharmaceutically acceptable salt" refers to a salt prepared from a compound of the present invention and a relatively nontoxic, pharmaceutically acceptable acid or base. When the compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting a neutral form of such compound with a sufficient amount of a pharmaceutically acceptable base in a pure solution or a suitable inert solvent. When the compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting a neutral form of such compound with a sufficient amount of a pharmaceutically acceptable acid in a pure solution or a suitable inert solvent.

[0025] The term "pharmaceutical excipients" refers to all substances contained in pharmaceutical preparations other than active pharmaceutical ingredients, which are generally divided into two categories: excipients and additives. Please refer to Part IV of the Pharmacopoeia of the People's Republic of China (2020 Edition), or Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009 Sixth Edition).

[0026] The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) ameliorating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade leading to or causing the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0027] The term "prevention" refers to a period of time in which a subject remains healthy relative to a disease or condition mentioned herein. It should be understood that the period depends on the amount of the drug compound that has been administered and the individual factors of the subject discussed elsewhere in this specification. It should be understood that prevention may not be effective in all subjects treated with compounds according to the present invention. However, the term requires that a statistically significant portion of a cohort or group of subjects is preferably effectively prevented from suffering from a disease or condition referred to herein or its associated symptoms. Preferably, in this case, it is envisioned that a group or group of subjects, which will usually develop a disease or condition referred to herein, i.e., without taking preventive measures according to the present invention. Those skilled in the art can use various well-known statistical evaluation tools discussed elsewhere in this specification to immediately determine whether a part is statistically significant.

[0028] The reagents and raw materials used in the present invention are commercially available.

[0029] The positive progressive effect of the present invention is that the nitrogen-containing heterocyclic compounds provided by the present invention have one or more of the following advantages: novel structure, high drug-like properties and excellent metabolic properties. DETAILED DESCRIPTION

[0030] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0031] Synthesis of Intermediate A

[0032] Synthesis route:

[0033]

[0034] first step

[0035] A-1 (3.0 g, 10.3 mmol) was dissolved in methanol (10 mL), cooled to 0 °C, sodium methoxide (630 mg, 11.4 mmol) was added, and stirred at room temperature for 18 hours. Water (20 ml) was added to the reaction solution for dilution, and dichloromethane (20 mL × 3) was used for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product containing the target compound, which was purified by silica gel column chromatography (petroleum ether / ethyl acetate, 3 / 2, v / v) to obtain A-2. 1 H NMR (400MHz, Chloroform-d) δ 4.68-4.52 (m, 4H), 4.06 (s, 3H), 1.52 (s, 9H). ESI-MS calculated value [M+H]+ = 286.1, measured value 286.0.

[0036] Step 2

[0037] A-2 (1.6 g, 5.60 mmol), 1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (460 mg, 0.56 mmol), potassium carbonate (2.32 g, 16.8 mmol), A-3 (1.41 g, 11.20 mmol) and 1,4-dioxane (10 mL) and water (1 mL) were heated to 90 °C and stirred for 18 hours under nitrogen protection. The reaction solution was cooled to room temperature, filtered, and the filtrate was diluted with water (20 mL), extracted with ethyl acetate (20 mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product containing the target compound, which was purified by silica gel column chromatography (petroleum ether / ethyl acetate, 1 / 4, v / v) to obtain A-4. 1 H NMR (400 MHz, Chloroform-d) δ 4.68-4.49 (m, 4H), 4.02 (s, 3H), 2.65 (s, 3H), 1.51 (s, 9H). ESI-MS calculated value [M+H] + =266.1, measured value 266.1.

[0038] Step 3

[0039] Dissolve A-4 (1.1 g, 4.15 mmol) in dichloromethane (10 mL), add trifluoroacetic acid (10 mL), and stir at room temperature for 3 hours. The reaction solution is concentrated under reduced pressure to obtain the trifluoroacetate salt of intermediate A. ESI-MS theoretical calculated value [M+H] + = 166.1, measured value 166.1.

[0040] Example 1

[0041] Synthesis of compound 1

[0042] Synthesis route:

[0043]

[0044] first step

[0045] 1-1 (2.42 g, 5.59 mmol, containing 66% trifluoroacetic acid), 2-1 (1 g, 5.32 mmol), cesium carbonate (10.4 g, 31.9 mmol), P,P'-(9,9-dimethyl-9H-xanthene-4,5-diyl)bis[N,N,N',N'-tetraethyl-phosphinate diamide] (297 mg, 0.53 mmol) and tris(dibenzylideneacetone)dipalladium (487 mg, 0.53 mmol) were added to 1,4-dioxane (25 mL) and stirred at 100°C for 16 hours under nitrogen protection. The reaction solution was cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL×3), the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether / ethyl acetate, 5 / 1, v / v) to obtain 2-2. 1 H NMR (400 MHz, DMSO-d6): δ 7.74 (d, J = 5.6 Hz, 1H), 6.05(dd, J = 5.6, 2.0 Hz, 1H), 5.62 (d, J = 2.0 Hz, 1H), 4.10-3.97 (m, 4H), 3.77(s, 3H), 3.60-3.50 (m, 2H), 3.06-2.94 (m, 1H), 2.70 (d, J = 7.6 Hz, 2H), 1.18(t, J = 7.2 Hz, 3H). ESI-MS calculated value [M+H] + = 251.14, measured value 251.1.

[0046] Step 2

[0047] Dissolve 2-2 (700 g, 2.80 mmol) in tetrahydrofuran (7 mL) and water (2 mL), cool to 0°C, add lithium hydroxide monohydrate (120 mg, 2.94 mmol), and stir at room temperature for 16 hours. After the reaction, adjust the pH to 3-4 with 1 mol / L dilute hydrochloric acid, concentrate under reduced pressure, and purify by high performance liquid chromatography (C18 spherical, 20-35µm, 100A, 80g, mobile phase: acetonitrile-0.1% formic acid aqueous solution, gradient: 10-32%, retention time: 13-19min) to obtain 2-3. 1H NMR (400 MHz, DMSO-d6): δ 12.36 (br s, 1H), 7.82 (d, J =7.2 Hz, 1H), 6.31 (dd, J = 7.2, 2.0 Hz, 1H), 6.02 (d, J = 2.0 Hz, 1H), 4.35-4.25 (m, 2H), 3.99 (s, 3H), 3.93-3.83 (m, 2H), 3.12-3.03 (m, 1H), 2.71 (d, J= 7.6 Hz, 2H). ESI-MS calculated value [M+H] + = 223.11, measured value 223.0.

[0048] Step 3

[0049] Intermediate A (122 mg, 0.74 mmol) was dissolved in N,N-dimethylformamide (3 mL), 2-3 (150 mg, 0.67 mmol) and N,N-diisopropylethylamine (866 mg, 6.70 mmol) were added at 0°C, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (280 mg, 0.74 mmol) was added after 30 minutes. The mixture was heated to 25°C and stirred for 2 hours. After the reaction was completed, water (20 mL) was added for dilution, and ethyl acetate (20 mL×3) was used for extraction. The organic layers were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The final product, compound 1, was purified by high performance liquid chromatography (Waters 3767 / Qda Column: Agilent C18, 19*250 mm, 10 um; mobile phase: acetonitrile-0.1% formic acid aqueous solution, gradient: 12-22%, retention time: 6.0-9.0 min). 1H NMR (400 MHz, DMSO-d6): δ 7.74 (d, J = 5.6 Hz, 1H), 6.05 (dd, J = 5.6, 2.0 Hz,1H), 5.62 (d, J = 1.6 Hz, 1H), 4.80-4.70 (m, 2H), 4.56-4.45 (m, 2H), 4.08-3.98 (m, 2H), 3.96 (s, 3H), 3.74 (s, 3H), 3.59-3.53 (m, 2H), 3.10-3.00 (m,1H), 2.81 (d, J = 7.6 Hz, 2H), 2.55 (s, 3H). ESI-MS calculated value [M+H] + = 370.19, measured value 370.3.

[0050] Activity Test Example 1 Evaluation of the PAM activity of compounds on M4 receptors

[0051] Purpose:

[0052] The activity of the compounds on the M4 receptor was determined using the FLIPR CALCIUM 6 ASSAYKIT kit using a stably transfected cell line expressing the human M4 receptor (M4-Gα15-CHO).

[0053] Experimental Materials:

[0054]

[0055] Experimental equipment:

[0056]

[0057] Cell treatment:

[0058] 1. This study used a CHO cell line that stably expresses the M4-Gα15 receptor, expressing the human CHRM4 and GNA15 genes respectively;

[0059] 2. Cell treatment: M4-Gα15-CHO cells were cultured in F-12 medium containing 10% fetal bovine serum, 0.4 mg / mL hygromycin B, and 0.8 mg / mL geneticin G418 at 37°C and 5% carbon dioxide. Remove the old medium and wash once with phosphate buffer, then add 1 mL of trypsin solution and incubate at 37°C for about 2 minutes. When the cells detach from the bottom of the dish, add about 5 mL of complete medium preheated at 37°C. Gently blow the cell suspension with a pipette to separate the aggregated cells. Transfer the cell suspension to a sterile centrifuge tube and centrifuge for 5 minutes (1000 rpm).

[0060] Experimental operation:

[0061] 1. Cell plating: Digest and collect M4-Gα15-CHO cells, resuspend and count them, and then inoculate them into 384-well cell plates. The inoculation density is: 1.2×10 4 cells / 25 μL / well. Then place the cell plate in a 37°C, 5% CO2 incubator for about 20 hours;

[0062] 2. After 24 hours, prepare the loading buffer according to the instructions of the FLIPR CALCIUM 6 ASSAY KIT kit: freeze and thaw Component A to room temperature, dilute it with detection buffer and probenecid solution to obtain the loading buffer, and keep it at room temperature for use;

[0063] 3. Remove the culture medium from the cell plate, quickly add 35 μL of loading buffer to each well, centrifuge and incubate the cell plate at 37 °C in the dark for 120 minutes;

[0064] 4. Prepare the working solution of the compound to be tested, transfer 5 μL to the corresponding cell wells, and incubate at 37 °C in the dark for 30 minutes;

[0065] 5. Prepare 30 nM acetylcholine agonist working solution and transfer 20 μL / well to the 384-well sample plate;

[0066] 6. Place the cell plate, sample plate and pipette tip in the corresponding position of the FLIPR instrument, add 10 μL of the agonist diluted in step 5 to each experimental well using FLIPR, and collect data at a wavelength of 515 nm-575 nm;

[0067] 7. Plot the signal value against the compound concentration and use the GraphPad Prism software nonlinear regression method to perform curve fitting and EC 50 calculate.

[0068] Preparation of detection buffer and loading buffer:

[0069] 1. Mix 0.5 mol / L 2-(4-(2-hydroxyethyl)piperazine)ethanesulfonic acid buffer and Hank's balanced salt solution (pH 7.4) at a volume ratio of 1:24 to prepare the detection buffer. Add 10 mL of the detection buffer to a bottle of lyophilized powder CALCIUM 6 ASSAY KIT Component A and store in aliquots at -20°C.

[0070] 2. Dissolve probenecid powder in 1 mol / L sodium hydroxide solution to a final concentration of 250 mol / L. Mix the detection buffer, frozen-thawed Component A, and probenecid solution in a volume ratio of 44:5:1 to prepare the loading buffer.

[0071] Experimental results:

[0072]

[0073] Experimental conclusion:

[0074] The experimental samples (compounds) were prepared from the corresponding examples, and the results are shown in the table above. The compounds of the present application showed agonist activity on the M4 receptor in this test system.

[0075] Activity Test Example 2 Evaluation of the Brain Penetration Properties of Compounds

[0076] Purpose:

[0077] The brain tissue / plasma partition coefficients of the compounds obtained in the examples of the present invention, the compounds in CN117946112A and WO2024130068A1 in CD-1 mice were evaluated.

[0078] Experimental Materials:

[0079]

[0080] Experimental procedures:

[0081] The candidate compound was prepared into a clear solution or suspension with a designated solvent and administered orally to three CD-1 mice. The solvent for oral administration was 5% dimethyl sulfoxide + 95% (10% polyethylene glycol (15)-hydroxystearate aqueous solution, and the drug concentration was 0.5 mg / ml. Whole blood samples and brain tissue samples were collected 1 hour after administration into commercial EDTA2K anticoagulant tubes, centrifuged to obtain upper plasma and brain tissue homogenate samples, added acetonitrile solution containing internal standard to precipitate protein, centrifuged to obtain the supernatant, added an equal volume of water, centrifuged again, and the supernatant was sampled. The drug concentration in plasma samples and brain tissue samples was quantitatively analyzed by LCMS / MS analysis method.

[0082] Experimental methods:

[0083]

[0084] Experimental results:

[0085]

[0086]

[0087] Experimental conclusion:

[0088] The test sample is prepared according to the corresponding embodiment, or prepared by the preparation method disclosed in CN117946112A and WO2024130068A1.

[0089] The results show that the compound of the present application has a larger brain-blood partition coefficient, indicating that it has good brain-penetrating properties and has better therapeutic effects. For neurological diseases, the drug target is located in the brain, and the better the brain-penetrating ability of the molecule, generally speaking, the better the therapeutic effect.

[0090] .

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, characterized in that: The compound of formula I is 2. A pharmaceutical composition comprising the compound of formula I or a pharmaceutically acceptable salt thereof as claimed in claim 1, and a pharmaceutical excipient.

3. Use of the compound of formula I according to claim 1, a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 2 in the preparation of an M4 muscarinic acetylcholine receptor agonist.

4. Use of the compound of formula I according to claim 1, a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 2 in the preparation of a medicament for treating and / or preventing a mental illness; the mental illness is major depressive disorder, bipolar disorder or schizophrenia.

5. Use of a compound of formula I as claimed in claim 1, a pharmaceutically acceptable salt thereof or a pharmaceutical composition as claimed in claim 2 in the preparation of a medicament for treating and / or preventing a disease mediated by muscarinic acetylcholine receptors; The muscarinic acetylcholine receptor mediated disease is an M4 muscarinic acetylcholine receptor mediated disease.

6. The use according to claim 5, characterized in that The M4 muscarinic acetylcholine receptor-mediated disease is Alzheimer's disease, schizophrenia, major depressive disorder or bipolar disorder.

Images