Sulfoxide imine compound and application thereof

Abstract

The invention provides a sulfoximine compound and application thereof. The structure of the sulfoximine compound is shown as a formula I: # imgabs0 # formula I. The invention further provides a preparation method of the sulfoximine compound. The compound provided by the invention shows good inhibitory activity, oral absorption and brain permeability on LRRK2 protein kinase, and meets the requirements of development of drugs for neurodegenerative diseases.

Description

Technical Field

[0001] The invention belongs to the field of medicinal chemistry, and specifically relates to a sulfoximine compound and application thereof, and in particular to a sulfoximine compound with good inhibitory effect and application thereof. Background Art

[0002] Leucine-rich repeat kinase 2 (LRRK2) is the protein product of the LRRK2 gene and is a serine / threonine protein kinase. Although the function of LRRK2 protein is not fully understood at present, LRRK2 gene mutations were found to be associated with Parkinson's disease (PD) as early as 2004. LRRK2 gene mutations are common in the kinase active region G2019S and I2020T and the GTPase region R1441G / C / H and Y1699C mutations, all of which can lead to increased LRRK2 protein kinase activity and induce a variety of pathological changes, including increased synuclein and TAU protein phosphorylation, nervous system inflammation, and neuronal mitochondrial dysfunction.

[0003] Parkinson's disease (PD) is the second largest neurodegenerative disease after Alzheimer's disease, with more than 10 million patients worldwide, but no effective treatment has been found so far. 10% of PD patients have a clear family history, of which LRRK2 mutation is the most common cause. This mutation occurs in about 5% of familial PD and 1% of non-familial PD, and the clinical manifestations caused by these mutations are no different from those of sporadic PD. In addition, LRRK2 mutations can also lead to pathological changes in TAU protein, and pathological changes in TAU are a major feature of Alzheimer's disease. LRRK2 is believed to be located in the upstream stage of the pathogenesis of neurodegenerative diseases and also plays an important role in other neurodegenerative diseases. Therefore, the development of LRRK2-specific inhibitors has become one of the effective ways to treat PD and other neurodegenerative diseases.

[0004] Existing LRRK2 protein kinase compounds are not satisfactory in terms of enzyme inhibition activity, or have poor brain penetration, and cannot meet the requirements of drug development. Therefore, there is still an urgent need in the prior art for compounds with good protein kinase inhibition activity and / or good brain penetration. Summary of the invention

[0005] In view of the deficiencies of the prior art, the object of the present invention is to provide a sulfoximine compound and its application, in particular to provide a sulfoximine compound and its application with good inhibitory effect. The compound provided by the present invention exhibits good inhibitory activity, oral absorption and brain permeability against LRRK2 protein kinase, meeting the requirements of drug development for neurodegenerative diseases.

[0006] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides a sulfoximine compound or a stereoisomer, a tautomer, an isotope-labeled substance or a pharmaceutically acceptable salt thereof, wherein the structure of the sulfoximine compound is shown in Formula I:

[0008]

[0009] Wherein, A is selected from hydrogen, substituted or unsubstituted any one of the following groups: C 1～6 Alkyl, C 3～8 Cycloalkyl, C 6～14 aryl, 5-12 membered heteroaryl, 3-8 membered heterocyclic group, 5-10 membered bridged ring group or 5-12 membered spirocyclic group, wherein the substituents substituted are selected from C 1～4 Alkyl, C 1～6 Alkoxy, C 3～6 Cycloalkyl, 3- to 8-membered heterocyclic group, C 1～3 Alkylphosphoryl, C 1～3 Alkylsulfonyl, oxo, cyano or halogen.

[0010] L is selected from -H, -O-, -NH-, -S- or -N(C 1～4 When L is -H, A is absent.

[0011] R 1 Selected from hydrogen, cyano, hydroxyl, C 3～6 Cycloalkyl, substituted or unsubstituted C 1～6 Alkyl, substituted or unsubstituted C 1～6 Alkoxy, wherein the substituted substituent is selected from hydroxy, methylamino, dimethylamino, methoxy, carboxyl, oxo, cyano or halogen.

[0012] R 2 Selected from C 1～4 Alkyl, C 1～6 Alkoxy, C 3～6 Cycloalkyl, cyano or halogen.

[0013] m is selected from 0, 1 or 2. When m is 0, the substituent is absent. When m is 2, two R 2 Same or different.

[0014] X is selected from CH or N; Y and Z are independently selected from C or N.

[0015] R a is selected from hydrogen, cyano, halogen, C 1～3 Alkyl, C 1～3 Alkylamino, C 1～3 Alkylphosphoryl or C 1～3Alkylsulfonyl.

[0016] R b Selected from hydrogen, amino or C 1～3 Alkylamino.

[0017] R a and R b It can exist independently or be linked together with the carbon atoms to form a 5- to 12-membered heteroaromatic ring or a 5- to 8-membered heterocyclic ring.

[0018] The above-mentioned sulfoximine compounds with specific structures exhibit good inhibitory activity, oral absorption and brain permeability against LRRK2 protein kinase, and can achieve effective treatment of LRRK2 protein kinase-related diseases.

[0019] Preferably, the structure of the sulfoximine compound is as shown in Formula II:

[0020]

[0021] Among them, A, L, R 1 , R 2 , m, X, Y, Z, and R a The same limitations as above apply.

[0022] Preferably, the structure of the sulfoximine compound is as shown in Formula III:

[0023]

[0024] Among them, A, L, R 1 , R 2 , m, X, Y and Z have the same limited ranges as above.

[0025] M is selected from C or N. When M is selected from C, R 3 is selected from hydrogen or halogen; when M is selected from N, R 3 Does not exist.

[0026] Preferably, the structure of the sulfoximine compound is as shown in Formula IV:

[0027]

[0028] Among them, A, L, R 1 , R 2 , m, X, Y and Z have the same limited ranges as above.

[0029] R 4 , R 5 are independently selected from hydrogen or fluorine, R 6 Selected from hydrogen or C 1～3 alkyl.

[0030] Preferably, A is selected from the following structures:

[0031]

[0032] Preferably, said L is selected from -O- or -NH-.

[0033] Preferably, the sulfoximine compound is selected from any one of the following structures:

[0034]

[0035]

[0036]

[0037]

[0038]

[0039] In a second aspect, the present invention provides use of the above-mentioned sulfoximine compound or its stereoisomer, tautomer, isotope-labeled substance or pharmaceutically acceptable salt in the preparation of LRRK2 protein kinase inhibitors.

[0040] The above-mentioned LRRK2 protein kinase inhibitor can be used as a drug for LRRK2 protein kinase-related diseases, and can also be used as an inhibitor in scientific research, such as research on related signal pathways.

[0041] In a third aspect, the present invention provides a pharmaceutical composition comprising at least one sulfoximine compound as described above or a stereoisomer, tautomer, isotope-labeled substance or a pharmaceutically acceptable salt thereof.

[0042] In a fourth aspect, the present invention also provides the use of the above-mentioned sulfoximine compound or its stereoisomer, tautomer, isotope-labeled substance or pharmaceutically acceptable salt or the above-mentioned pharmaceutical composition in the preparation of a drug for treating and / or preventing LRRK2 protein kinase-related diseases.

[0043] Preferably, the disease comprises a neurodegenerative disease, an immune-related disease or a cancer, specifically comprising Parkinson's disease, Alzheimer's disease, Huntington's disease, Crohn's disease, inflammatory bowel disease, colitis, tuberculosis, leprosy, melanoma, non-melanoma skin cancer or breast cancer.

[0044] Compared with the prior art, the present invention has the following beneficial effects:

[0045] The present invention provides a sulfoximine compound with a specific structure, which exhibits good inhibitory activity, oral absorption and brain permeability against LRRK2 protein kinase, and can achieve effective treatment of LRRK2 protein kinase-related diseases. DETAILED DESCRIPTION

[0046] In order to further explain the technical means and effects adopted by the present invention, the technical solution of the present invention is further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0047] The term "alkyl" is used to denote a straight or branched chain saturated hydrocarbon group, for example, C 1～3 Alkyl refers to a saturated hydrocarbon group containing 1 to 3 carbon atoms. 1～4 Alkyl refers to a saturated hydrocarbon group containing 1 to 4 carbon atoms. 1～6 The alkyl group is a saturated hydrocarbon group containing 1 to 6 carbon atoms, including straight-chain, branched or cyclic alkyl groups. The alkyl group herein is preferably C 1～10 Alkyl, C 1～8 Alkyl, C 1～6 Alkyl or C 1～4 alkyl.

[0048] The term "alkoxy" refers to an alkyl-O- group. 1～6 "Alkoxy" is intended to include C 1 , C 2 , C 3 , C 4 , C 5 , C 6 Alkoxy. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), and tert-butoxy. Examples of alkoxy include methoxy, ethoxy, propoxy (e.g., n-propoxy, isopropoxy, and cyclopropoxy), tert-butoxy, and the like. Preferred alkoxy groups herein include C 1～10 Alkoxy, C 1～8 Alkoxy, C 1～6 Alkoxy and C 1～4 Alkoxy.

[0049] The term "cycloalkyl" refers to a cyclized alkyl group based on a non-aromatic carbocyclic ring. Cycloalkyl groups may include monocyclic, bicyclic or polycyclic ring systems. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc. 3～8 Cycloalkyl refers to a cycloalkyl group containing 3 to 8 carbon atoms. The preferred cycloalkyl group herein is C 3～8 Cycloalkyl, C 3～6 Cycloalkyl.

[0050] The term "heterocycloalkyl" or "heterocyclyl" refers to a non-aromatic heterocycloalkyl group in which 1 or 2 or 3 of the ring-forming carbon atoms are replaced by heteroatoms selected from O, N and S atoms. The heterocycloalkyl group preferably has 3, 4, 5, 6, 7 or 8 ring-forming atoms. Preferred heterocyclyl groups herein are C 3～8 Heterocyclic or C 3～6 Heterocyclic group.

[0051] "Aryl" refers to an aromatic carbocyclic group, including monocyclic, bicyclic, tricyclic or polycyclic aromatic hydrocarbons, such as phenyl, naphthyl, anthracenyl, phenanthrenyl, etc. Aryl is preferably a monocyclic, bicyclic or tricyclic ring system with 6 to 14 or 6 to 12 ring members. Preferably, at least one ring in the system is aromatic and each ring in the system contains 3 to 7 ring members. "Substituted aryl" means that at least one hydrogen atom on the phenyl ring of the aryl is replaced by a non-hydrogen moiety. The substituents of the aryl can be halogen, C 1～10 Alkoxy, -CN, -OH, -SH, -NH 2 , C 1～6 Alkyl. Preferred aryl groups include phenyl, biphenyl, indanyl, 1-naphthyl, 2-naphthyl and tetrahydronaphthyl. The term "aralkyl" or "arylalkyl" refers to an alkyl residue attached to an aryl ring. Non-limiting examples include benzyl, phenethyl, etc. The fused aryl group may be attached to another group at a suitable position on the cycloalkyl ring or the aromatic ring.

[0052] The term "heteroaromatic ring" or "heteroaryl" means a 5-12 membered aromatic monocyclic or aromatic bicyclic or aromatic polycyclic heterocyclic ring, which is fully unsaturated, partially unsaturated, and contains carbon atoms and 1, 2, 3 or 4 heteroatoms independently selected from N, O and S. The heteroaryl herein is preferably a 5-12 membered heteroaryl. The heteroaryl herein preferably has 1 or 2 or 3 heteroatoms independently selected from N, O and S. Preferably, the heteroaryl herein includes a 5-, 6- or 7-membered aromatic monocyclic ring or an 8-, 9-, 10-, 11- or 12-membered aromatic bicyclic or aromatic polycyclic heterocyclic ring; preferably, any heterocyclic ring defined above is fused to a benzene ring. The nitrogen and sulfur heteroatoms may be optionally oxidized. The nitrogen atom may be substituted or unsubstituted (i.e., N or NR, wherein R is H or, if defined, another substituent). The heterocyclic ring may be attached to its side group at any heteroatom or carbon atom that results in a stable structure. The heterocyclic groups described herein may be substituted on carbon or nitrogen atoms if the resulting compound is stable. The nitrogen in the heterocyclic ring may be optionally quaternized. Preferably, when the total number of S and O atoms in the heterocyclic ring exceeds 1, these heteroatoms are not adjacent to each other.Examples of aromatic heteroyl groups include, but are not limited to, acridinyl, azetidinyl, azinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiopyranyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, quinolinyl, decahydroquinolinyl, dihydrofuro[2,3-b]tetrahydrofuranyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, imidazopyridinyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isoquinolin ... benzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isothiazolopyridinyl, isoxazolyl, isoxazolopyridinyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolopyridinyl, oxazolidinyl, perylene, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl , 4-piperidinyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolopyridinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2-pyrrolidinone, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinazolinyl, quinuclidinyl, tetrazolyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl , thianthrenyl, thiazolyl, thiopyranyl, thiazolopyridinyl, thiopyranothiazolyl, thiopyranooxazolyl, thiopyranoimidazolyl, thiopyranyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and xanthenyl, quinolyl, isoquinolyl, phthalazinyl, quinazolinyl, indolyl, 1H-indazolyl, benzimidazolyl, 1,2,3,4-tetrahydroquinolyl, 1,2,3,4-tetrahydroisoquinolyl, 5,6,7,8-tetrahydro-quinolyl, 2,3-dihydro-benzofuranyl, 1,2,3,4-tetrahydro-quinazolinyl and 1,2,3,4-tetrahydro-quinazolinyl. The term "heteroaryl" may also include a biaryl structure formed by the above-defined "aryl" and a monocyclic "heteroaryl", such as but not limited to "-phenylbipyridyl-", "-phenylbipyrimidyl-", "-pyridylbiphenyl-", "-pyridylbipyrimidyl-", "-pyrimidylbiphenyl-".

[0053] The term "C1～3 "Alkylphosphoryl" refers to C 1～3 In the alkyl-substituted phosphoryl group, the two alkyl groups on the phosphoryl group may be the same or different.

[0054] The term "C 1～3 "Alkylsulfonyl" refers to C 1～3 Alkyl substituted sulfonyl.

[0055] The term "halogen" includes fluorine, chlorine, bromine and iodine.

[0056] "Bridged ring group" refers to a polycyclic group with 5 to 18 members, preferably 5 to 10 members, containing two or more cyclic structures, sharing two carbon atoms that are not directly connected to each other. More preferably, it is a 6 to 10-membered, and even more preferably a 7 to 10-membered bridged ring group. According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, pyridone or polycyclic bridged heterocyclic group, preferably a bicyclic, tricyclic or pyridone, and more preferably a bicyclic or tricyclic. The bridged ring group herein allows 1, 2 or 3 of the ring atoms to be selected from N, O, S, SO and / or S(O) 2 of heteroatoms.

[0057] "Spirocyclyl" refers to a polycyclic group of 5 to 18 members, preferably 6 to 12 members, containing two or more cyclic structures, sharing one carbon atom. According to the number of constituent rings, it can be divided into bicyclic and tricyclic, preferably bicyclic. The spirocyclyl herein allows one, two or three of the ring atoms to be selected from N, O, S, SO and / or S(O) 2 of heteroatoms.

[0058] The term "pharmaceutically acceptable salt" or "pharmaceutically acceptable salt" as used herein refers to salts that enable the compounds of the present invention to maintain their original biological activity and are suitable for medical use. The pharmaceutically acceptable salt of the compound represented by formula (I) can be a salt formed by a carboxyl group or an amine group (mainly an amine group) and a suitable base or acid, such as a salt formed with a suitable base (including a metal salt, an ammonium salt), or a salt formed with a suitable acid. The compound of the present invention preferably forms a salt with a suitable acid, and the acid can be selected from: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, trifluoroacetic acid and aspartic acid, preferably methanesulfonic acid or hydrochloric acid.

[0059] As used herein, the term "treat," ...

[0060] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutical substance, composition or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc, magnesium, calcium or zinc stearate or stearic acid) or solvent encapsulating substance, which is involved in carrying or transporting the subject compound from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.

[0061] The term "pharmaceutical composition" means a composition comprising a compound of the present invention and at least one other pharmaceutically acceptable carrier. "Pharmaceutically acceptable carrier" refers to a medium generally accepted in the art for delivering biologically active agents to animals (particularly mammals), including (i.e.) adjuvants, excipients or vehicles, such as diluents, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, aromatics, antibacterial agents, antifungal agents, lubricants and dispersants, depending on the nature of the mode of administration and dosage form.

[0062] The term “TLC” refers to thin layer chromatography;

[0063] In order to better illustrate the technical means and effects adopted by the present invention, the present invention is further described below in conjunction with non-limiting examples. The embodiments of the present invention, including the description provided in the embodiments, are intended to illustrate the embodiments of the present invention, and are not intended to limit the scope of any protection of the present invention. According to the present invention, it will be understood by those skilled in the art that many changes can be made to the disclosed specific embodiments without departing from the spirit and scope of the present invention and still the same or similar results can be obtained.

[0064] Unless otherwise stated, all materials / reagents were obtained from commercial suppliers and used without further purification. The structures of the compounds in the following examples were characterized and confirmed by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS).

[0065] 1 HNMR spectra were recorded on a Bruker Avance 400 MHz spectrometer at room temperature in deuterated dimethyl sulfoxide (d-DMSO), deuterated chloroform (CDCl 3 ), deuterated methanol (CD 3 OD) or deuterated water (D 2 O). Chemical shift values ​​(δ) are in ppm, using tetramethylsilane (TMS) or residual solvent peak as internal standard, and coupling constants (J) are in Hertz (Hz). 1The abbreviations for the multiplicity of peaks in HNMR spectra are as follows: s (singlet), d (doublet), t (triplet), q (quartet), qn (quintet), m (multiplet), br (broad).

[0066] The instrument used for liquid chromatography-mass spectrometry (LC-MS) is Shimadzu LCMS-2020, and the instrument used for preparative high performance liquid chromatography (Prep-HPLC) is Bonna-Agela FLEXA FL-H100G. The thin layer chromatography silica gel plate used for thin layer chromatography (TLC) is Yantai Huanghai HSGF254 thin layer chromatography silica gel plate. The specifications used for reaction monitoring are 2.5×8cm, and the coating thickness is 0.2±0.03mm. The specifications used for separation and purification are 20×20cm, and the coating thickness is 0.4-0.5mm. The silica gel column chromatography method uses Qingdao Marine Silica Gel 100-200 mesh or 200-300 mesh silica gel as the carrier.

[0067] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any method and material similar or equivalent to the described content can be applied to the method of the present invention.

[0068] Example 1

[0069] (1s,4s)-4-(4-((4-(((1r,4r)-4-hydroxy-4-methylcyclohexyl)oxy)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)-1-(methylimino)hexahydro-1λ 6 -Thiopyran 1-oxide

[0070]

[0071] Implementation steps (implementation method 1):

[0072]

[0073] Step 1: Synthesis of Intermediate 1-c: 4-(4-nitrophenyl)-3,6-dihydro-2H-thiopyran

[0074]

[0075] Tetrakis(triphenylphosphine)palladium (487 mg, 0.42 mmol) was added to a solution of 1,4-dioxane (10 mL) and water (2.5 mL) containing 1-iodo-4-nitrobenzene (1-a, 1.05 g, 4.21 mmol), 2-(3,6-dihydro-2H-thiopyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (1-b, 1.0 g, 4.42 mmol) and sodium carbonate (894 mg, 8.42 mmol). The mixture was reacted at 90°C under a nitrogen atmosphere for 16 hours. Stirring was stopped after monitoring the progress of the reaction by TLC. After the mixture was cooled to room temperature, ethyl acetate (20 ml) and water (10 ml) were added to dilute it, and the organic phase was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether containing 5% ethyl acetate as eluent) to give the intermediate 4-(4-nitrophenyl)-3,6-dihydro-2H-thiopyran as a yellow solid (1-c, 590 mg, 60%).

[0076] 1 HNMR (400 MHz, CDCl 3 ): δ8.20-8.17 (m, 2H), 7.50-7.47 (m, 2H), 6.37-6.34 (m, 1H), 3.40-3.37 (m, 2H), 2.91 (t, J=5.8Hz, 2H), 2.74-2.70 (m, 2H).

[0077] Step 2: Synthesis of Intermediate 1-d: 4-(4-nitrophenyl)-3,6-dihydro-2H-thiopyran 1-oxide

[0078]

[0079] Sodium periodate (572 mg, 2.67 mmol) was added in batches to a solution of methanol (12 ml) and water (3 ml) containing 4-(4-nitrophenyl)-3,6-dihydro-2H-thiopyran (1-c, 590 mg, 2.67 mmol) under ice bath conditions. The mixture was gradually warmed to room temperature and reacted for 8 hours. Stirring was stopped after monitoring the progress of the reaction by TLC. The mixture was filtered through diatomaceous earth and the filtrate was collected. The filter cake was washed with methanol (10 mL × 2) and the filtrate was combined. After concentration under reduced pressure, it was diluted with water (10 ml) and dichloromethane (10 ml x 2), the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was slurried with a petroleum ether solution containing 10% ethyl acetate to obtain the intermediate 4-(4-nitrophenyl)-3,6-dihydro-2H-thiopyran 1-oxide as a light yellow solid (1-d, 540 mg, 85%);

[0080] LC-MS:(ES +):m / z 238.10[M+H] + .

[0081] 1 HNMR (400 MHz, CDCl 3 ): δ8.22 (d, J=8.8Hz, 2H), 7.54 (d, J=8.8Hz, 2H), 6.12-6.10 (m, 1H), 3.6 4-3.51 (m, 2H), 3.33-3.17 (m, 2H), 2.95-2.88 (m, 1H), 2.79-2.72 (m, 1H).

[0082] Step 3: Synthesis of intermediate 1-e: (1s, 4s)-4-(4-aminophenyl)tetrahydro-2H-thiopyran 1-oxide

[0083]

[0084] Platinum dioxide (100%, 300 mg) was added to a methanol (10 ml) solution containing 4-(4-nitrophenyl)-3,6-dihydro-2H-thiopyran 1-oxide (1-d, 300 mg, 1.26 mmol), and the reaction mixture was stirred under a hydrogen atmosphere at room temperature for 18 hours. Stirring was stopped after monitoring the progress of the reaction by TLC. The reaction mixture was filtered through diatomaceous earth and the filtrate was collected. The filter cake was washed with methanol (10 mL × 2) and the filtrate was combined and concentrated under reduced pressure to obtain a crude product (1s, 4s)-4-(4-aminophenyl)tetrahydro-2H-thiopyran 1-oxide as a yellow solid (1-e, 200 mg, 75%).

[0085] 1 HNMR (400MHz, d-DMSO): δ6.88 (d, J=8.4Hz, 2H), 6.51 (d, J=8.4Hz, 2H), 4.87 (s, 2H), 2.93- 2.87 (m, 2H), 2.76-2.68 (m, 2H), 2.59-2.54 (m, 1H), 2.25-2.14 (m, 2H), 1.66-1.62 (m, 2H).

[0086] Step 4: Synthesis of Intermediate 1-f: tert-Butyl (4-((1s,4s)-1-oxytetrahydro-2H-thiopyran-4-yl)phenyl)carbamate

[0087]

[0088] Di-tert-butyl dicarbonate (521 mg, 2.39 mmol) was added dropwise to a solution of tetrahydrofuran (3 ml) and water (2 ml) containing compound (1s, 4s)-4-(4-aminophenyl)tetrahydro-2H-thiopyran 1-oxide (1-e, 200 mg, 0.95 mmol) and sodium carbonate (304 mg, 2.87 mmol) at room temperature, and the mixture was stirred at room temperature for 6 hours. Stirring was stopped after monitoring the progress of the reaction by TLC. Ethyl acetate (10 ml) and water (10 ml) were added to the reaction solution for dilution, and the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain crude product (tert-butyl 4-((1s, 4s)-1-oxidotetrahydro-2H-thiopyran-4-yl)phenyl)carbamate as a light yellow solid (1-f, 175 mg, 59%).

[0089] 1 HNMR (400MHz, d-DMSO): δ9.26 (s, 1H), 7.38 (d, J=8.4Hz, 2H), 7.12 (d, J=8.8Hz, 2H), 2. 98-2.88(m, 2H), 2.79-2.65(m, 3H), 2.33-2.20(m, 2H), 1.73-1.62(m, 2H), 1.47(s, 9H).

[0090] Step 5: Intermediate 1-g: (4-((1s,4s)-1-Imino-1-oxyhexahydro-1λ 6 Synthesis of tert-butyl (4-thiopyran-4-yl)phenyl)carbamate

[0091]

[0092] To a methanol (1 mL) solution containing tert-butyl (4-((1s,4s)-1-oxytetrahydro-2H-thiopyran-4-yl)phenyl)carbamate (1-f, 120 mg, 0.39 mmol) and ammonium carbamate (121 mg, 1.55 mmol) was added iodobenzene diacetate (375 mg, 1.15 mmol). The mixture was reacted at room temperature for 0.5 hours. After monitoring the progress of the reaction by TLC, stirring was stopped. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent solution containing 5% methanol in dichloromethane) to obtain (4-((1s,4s)-1-imino-1-oxyhexahydro-1λ 6 -butyl thiopyran-4-yl)phenyl)carbamate was obtained as a white solid (1-g, 82 mg, 65%).

[0093] 1HNMR (400MHz, d-DMSO): δ9.27 (s, 1H), 7.37 (d, J=8.4Hz, 2H), 7.11 (d, J=8.4Hz, 2H), 3.72 (s, 1 H), 3.17-3.10(m, 2H), 3.01-2.97(m, 2H), 2.82-2.75(m, 1H), 2.08-1.90(m, 4H), 1.46(s, 9H).

[0094] Step 6: Intermediate 1-h: (4-((1s,4s)-1-(methylimino)-1-oxohexahydro-1λ 6 Synthesis of tert-butyl (4-thiopyran-4-yl)phenyl)carbamate

[0095]

[0096] To the containing (4-((1s,4s)-1-imino-1-oxyhexahydro-1λ 6 To a solution of tert-butyl 2-((1-thiopyran-4-yl)phenyl)carbamate (1-g, 80 mg, 0.24 mmol), paraformaldehyde (paraformaldehyde, 59 mg, 1.97 mmol), and triethylsilane (86 mg, 0.74 mmol) in acetonitrile (5 ml) was added trifluoroacetic acid (87 mg, 0.76 mmol), and the reaction solution was stirred at room temperature for 16 hours. Stirring was stopped after monitoring the progress of the reaction by TLC. The mixture was distributed between ethyl acetate (10 ml) and saturated sodium carbonate solution (10 ml). The organic phase was collected, and the aqueous phase was extracted with ethyl acetate (10 ml x 2) and the organic phases were combined and washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative TLC (dichloromethane elution solution containing 5% methanol) to obtain (4-((1s, 4s)-1-(methylimino)-1-oxyhexahydro-1λ 6 tert-Butyl 4-thiopyran-4-yl)phenyl)carbamate was obtained as a white solid (1-h, 53 mg, 63%).

[0097] LC-MS:(ES + ):m / z 339.15[M+H] + .

[0098] 1 HNMR (400MHz, d-DMSO): δ9.27 (s, 1H), 7.38 (d, J = 8.4Hz, 2H), 7.12 (d, J = 8.4Hz, 2H), 3.24-3.1 4(m, 2H), 3.07-3.00(m, 2H), 2.87-2.77(m, 1H), 2.67(s, 3H), 2.07-1.94(m, 4H), 1.46(s, 9H).

[0099] Step 7: Fragment 1-i: (1s, 4s)-4-(4-aminophenyl)-1-(methylimino)hexahydro-1λ 6 -Synthesis of Thiopyran 1-oxide

[0100]

[0101] To the containing (4-((1s,4s)-1-imino-1-oxyhexahydro-1λ 6 To dichloromethane (2mL) of tert-butyl (1-h, 50mg, 0.15mmol) (1-thiopyran-4-yl)phenyl)carbamate (1-h, 50mg, 0.15mmol) was added trifluoroacetic acid (1mL), and the mixture was reacted at room temperature for 0.5 hours. Stirring was stopped after monitoring the progress of the reaction by TLC. The reaction solution was concentrated under reduced pressure, water (10ml) was added, the pH was adjusted to 8-9 with saturated sodium carbonate solution, and extracted with dichloromethane (10ml×2). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative TLC (dichloromethane elution solution containing 10% methanol) to obtain (1s,4s)-4-(4-aminophenyl)-1-(methylimino)hexahydro-1λ 6 -Thiopyran 1-oxide was obtained as a white solid (1-i, 33 mg, 94%).

[0102] LC-MS:(ES + ):m / z 239.15[M+H] + .

[0103] Step 8: Compound 1: (1s, 4s)-4-(4-((4-(((1r, 4r)-4-hydroxy-4-methylcyclohexyl)oxy)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)-1-(methylimino)hexahydro-1λ 6 -Synthesis of Thiopyran 1-oxide

[0104]

[0105] To the intermediate (1s,4s)-4-(4-aminophenyl)-1-(methylimino)hexahydro-1λ 6-Thiopyran 1-oxide (1-i, 30 mg, 0.12 mmol) and fragment (1r, 4r)-4-((2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)oxy)-1-methylcyclohexyl-1-ol (1-j, 47 mg, 0.15 mmol), 1,1'-bis(diphenylphosphino)ferrocene (14 mg, 0.025 mmol) and cesium carbonate (163 mg, 0.5 mmol) in 1,4-dioxane (1 ml) solution were added with tetrakis(triphenylphosphine)palladium (14.5 mg, 0.012 mmol), the reaction solution was replaced with nitrogen three times, heated to 110°C and stirred for 16 hours. After monitoring the progress of the reaction by TLC, stirring was stopped. After the reaction solution was concentrated under reduced pressure, the residue was diluted with ethyl acetate (10 mL) and saturated water (10 mL), the organic phase was collected, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative TLC (eluent containing 5% methanol in dichloromethane) to give (1s,4s)-4-(4-((4-(((1r,4r)-4-hydroxy-4-methylcyclohexyl)oxy)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenyl)-1-(methylimino)hexahydro-1λ 6 -Thiopyran 1-oxide was obtained as a white solid (Compound 1, 26 mg, 60%).

[0106] LC-MS:(ES + ):m / z 513.20[M+H] + .

[0107] 1 HNMR (400 MHz, CD 3 OD): δ8.35 (s, 1H), 7.61 (d, J=8.8Hz, 2H), 7.25 (d, J=8.4Hz, 2H), 5.43 (brs, 1H), 3.37-3.31 (m, 2H), 3.20-3.12 (m, 2H), 2 .94-2.87(m, 1H), 2.81(s, 3H), 2.31-2.17(m, 4H), 2.06-1.99(m, 2H), 1.86-1.71(m, 4H), 1.56-1.51(m, 2H), 1.22(s, 3H).

[0108] Example 2

[0109] 1-(Ethylimino)-4-(5-((4-(((1r,4r)-4-hydroxy-4-methylcyclohexyl)oxy)-5-(trifluoromethyl)pyrimidin-2-yl)amino)pyridin-2-yl)-1λ 6 -Thiomorpholine 1-oxide

[0110]

[0111] Implementation steps (implementation method 2):

[0112]

[0113] Step 1: Synthesis of Intermediate 2-b: 4-(5-nitropyridin-2-yl)thiomorpholine 1-oxide

[0114]

[0115] Sodium periodate (323 mg, 1.51 mmol) was added to a solution of anhydrous methanol (10 mL) containing 4-(5-nitropyridin-2-yl)thiomorpholine (2-a, 285 mg, 1.26 mmol). The mixture was stirred at room temperature for 5 hours. Stirring was stopped after monitoring the progress of the reaction by TLC. The reaction mixture was filtered through diatomaceous earth and the filtrate was collected. The filter cake was washed with methanol (10 mL × 2) and the filtrate was combined and concentrated under reduced pressure to obtain the crude product 4-(5-nitropyridin-2-yl)thiomorpholine 1-oxide as a yellow solid (2-b, 300 mg, 98%).

[0116] LC-MS:(ES + ):m / z 242.25[M+H] + .

[0117] 1 HNMR (400 MHz, CDCl 3 )δ9.09 (d, J=2.5Hz, 1H), 8.31 (dd, J=9.4, 2.8Hz, 1H), 6.72 (d, J=9.4Hz, 1H), 4. 46 (d, J=14.8Hz, 2H), 4.32–4.19 (m, 2H), 3.02–2.86 (m, 2H), 2.85–2.69 (m, 2H).

[0118] Step 2: Intermediate 2-c: 1-Imino-4-(5-nitropyridin-2-yl)-1λ 6 -Synthesis of Thiomorpholine 1-Oxide

[0119]

[0120] Iodobenzene diacetate (1.2 g, 3.72 mmol) was added to a methanol (10 mL) solution containing 4-(5-nitropyridin-2-yl)thiomorpholine 1-oxide (2-b, 300 mg, 1.24 mmol) and ammonium carbamate (290 mg, 3.72 mmol). The mixture was reacted at room temperature for 2.5 hours. Stirring was stopped after monitoring the progress of the reaction by TLC. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent solution containing 5% methanol in dichloromethane) to obtain 1-imino-4-(5-nitropyridin-2-yl)-1λ 6 -Thiomorpholine 1-oxide was obtained as a yellow solid (2-c, 256 mg, 81%).

[0121] LC-MS:(ES + ):m / z 257.00[M+H] + .

[0122] 1 HNMR (400 MHz, CDCl 3 ): δ8.44 (d, J=5.4Hz, 1H), 7.43 (d, J=1.4Hz, 1H), 7.40 (dd, J=5.4, 1.7Hz, 1H), 4.31 (ddd, J=14.4, 9.6, 4.8Hz, 2H), 4.18 (dt, J=11.0, 5.2Hz, 2H), 3.12 (d, J=5.2Hz, 4H).

[0123] Step 3: Intermediate 2-d: 1-(ethylimino)-4-(5-nitropyridin-2-yl)-1λ 6 -Synthesis of Thiomorpholine 1-Oxide

[0124]

[0125] To contain 1-imino-4-(5-nitropyridin-2-yl)-1λ 6-Thiomorpholine 1-oxide as a yellow solid (2-c, 256 mg, 1.00 mmol), acetaldehyde (220 mg, 5.00 mmol), triethylsilane (348 mg, 3.00 mmol) in acetonitrile (10 ml) solution was added with trifluoroacetic acid (342 mg, 3.00 mmol), and the reaction solution was stirred at room temperature for 16 hours. After monitoring the progress of the reaction by TLC, stirring was stopped. The mixture was distributed between ethyl acetate (10 ml) and saturated sodium carbonate solution (10 ml). The organic phase was collected, and the aqueous phase was extracted with ethyl acetate (10 ml x 2) and then the organic phases were combined and washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative TLC (dichloromethane elution solution containing 5% methanol) to obtain 1-(ethylimino)-4-(5-nitropyridine-2-yl)-1λ 6 -Thiomorpholine 1-oxide was obtained as a yellow solid (2-d, 260 mg, 91%).

[0126] LC-MS:(ES + ):m / z 285.10[M+H] + .

[0127] 1 HNMR (400 MHz, CDCl 3 )δ8.37 (d, J=5.4Hz, 1H), 7.36 (d, J=1.4Hz, 1H), 7.33 (dd, J=5.4, 1.7Hz, 1H), 4.22 (ddd, J=14.4, 7.1, 2.9Hz, 2H), 4.09 (d dd, J=14.6, 7.8, 2.5Hz, 2H), 3.23 (dd, J=15.1, 4.6Hz, 2H), 3.14 (q, J=7.2Hz, 2H), 3.10–3.01 (m, 2H), 1.22–1.18 (m, 3H).

[0128] Step 4: Intermediate 2-e: 4-(5-aminopyridin-2-yl)-1-(ethylimino)-1λ 6 -Synthesis of Thiomorpholine 1-Oxide

[0129]

[0130] To contain 1-(ethylimino)-4-(5-nitropyridin-2-yl)-1λ 6Pd / C (10%, 50 mg) was added to a solution of -thiomorpholine 1-oxide (2-d, 50 mg, 0.18 mmol) in methanol (4 ml), and the reaction mixture was stirred under a hydrogen atmosphere at room temperature for 12 hours. Stirring was stopped after monitoring the progress of the reaction by TLC. The reaction mixture was filtered through diatomaceous earth and the filtrate was collected. The filter cake was washed with methanol (10 mL × 2) and the filtrate was combined and concentrated under reduced pressure to obtain a crude product of 4-(5-aminopyridin-2-yl)-1-(ethylimino)-1λ 6 -Thiomorpholine 1-oxide was obtained as a yellow solid (2-e, 13 mg, 29%).

[0131] LC-MS:(ES + ):m / z 255.05[M+H] + .

[0132] Step 5: Compound 2: 1-(ethylimino)-4-(5-((4-(((1r,4r)-4-hydroxy-4-methylcyclohexyl)oxy)-5-(trifluoromethyl)pyrimidin-2-yl)amino)pyridin-2-yl)-1λ 6 -Synthesis of Thiomorpholine 1-Oxide

[0133]

[0134] To the intermediate 4-(5-aminopyridin-2-yl)-1-(ethylimino)-1λ 6 -Thiomorpholine 1-oxide (2-e, 12.8 mg, 0.05 mmol) and fragment (1r, 4r)-4-((2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)oxy)-1-methylcyclohexyl-1-ol (1-j, 15.5 mg, 0.05 mmol), 1,1'-bis(diphenylphosphino)ferrocene (5.6 mg, 12 μmol) and potassium carbonate (13.8 mg, 0.10 mmol) in 1,4-dioxane (1 ml) solution were added with tetrakis(triphenylphosphine)palladium (5.8 mg, 5 μmol), the reaction solution was replaced with nitrogen three times, heated to 100°C and stirred for 12 hours. After monitoring the progress of the reaction by TLC, stirring was stopped. After the reaction solution was concentrated under reduced pressure, the residue was diluted with ethyl acetate (10 mL) and saturated water (10 mL), the organic phase was collected, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative TLC (eluent containing 5% methanol in dichloromethane) to give 1-(ethylimino)-4-(5-((4-(((1r,4r)-4-hydroxy-4-methylcyclohexyl)oxy)-5-(trifluoromethyl)pyrimidin-2-yl)amino)pyridin-2-yl)-1λ 6 -Thiomorpholine 1-oxide was a light yellow solid (Compound 2, 26 mg, 60%).

[0135] LC-MS:(ES + ):m / z 529.25[M+H] + .

[0136] 1 H NMR (400 MHz, CDCl 3 )δ8.54 (d, J=0.8Hz, 1H), 8.37 (d, J=5.4Hz, 1H), 7.36 (d, J=1.4Hz, 1H), 7.33 (dd, J=5.4, 1.7Hz, 1H), 5.59-5.53 (m, 1H), 4.22 (ddd, J=14.4, 7.1, 2.9Hz, 2H), 4.09 (ddd, J=14.6, 7.8, 2.5Hz, 2H), 3.23 (dd, J=15.1, 4.6Hz, 2H), 3.14 (q, J=7.2Hz, 2H), 3.10-3.01 (m, 2H), 2.10-1.99 (m, 2H), 1.89 (dd, J=14.0, 4.2Hz, 2H), 1.80-1.71(m, 2H), 1.56-1.54(m, 2H), 1.30(s, 3H), 1.22-1.18(m, 3H).

[0137] The following compounds in Table 1 were synthesized using a similar method as described above:

[0138] Table 1

[0139]

[0140]

[0141]

[0142]

[0143]

[0144]

[0145]

[0146]

[0147] Biological activity test example:

[0148] Biological test example-1: LRRK2 kinase activity inhibition experiment:

[0149] The ADP-Glo ​​method was used to detect the inhibitory effect of the compound on the activity of LRRK2 kinase. The compound was diluted with 1× kinase buffer, and 3μL of 1mM compound DMSO storage solution was added to 57μL of 1× kinase buffer D, and the compound was diluted 5 times with 1× kinase buffer. 1μL of the compound diluted with kinase buffer was added to a Greiner 384-well plate, and a DMSO blank control group and a buffer background group were set up. Use 1× kinase buffer kinase solution (kinase concentration titration and S / N=10 to determine the kinase dilution ratio), add 2μL of 2.5 times the final concentration of kinase solution, centrifuge the plate at 1000g for 30 seconds, and place at room temperature for 15 minutes. Prepare a mixed solution of ATP and kinase substrate at 2.5 times the final concentration in 1× kinase buffer, add 2μL of 2.5 times the final concentration of ATP and kinase substrate, and start the reaction. Centrifuge the plate at 1000g for 30 seconds, seal the plate, and place at room temperature for 1 hour. Add 5 μL ADP-Glo ​​reagent and incubate at room temperature for 40 minutes; then add 10 μL detection reagent and incubate at room temperature for 40 minutes. Detect the luminescent signal value on the Biotek Synergy H1 microplate reader. Conversion inhibition rate % = 100-(compound signal value-background signal value) / (blank control signal value-background signal value)×100. The dose response is obtained by fitting the log[inhibitor] vs.response-Variable slope curve to obtain IC 50 value.

[0150] The IC values ​​of some compounds in the LRRK2 kinase activity assay are provided in Table 2 below: 50 Range:

[0151] (IC 50 Range: A: <0.1μM; B: 0.1-0.5μM; C: >0.5μM):

[0152] Table 2 IC of compounds in LRRK2 kinase activity assay 50

[0153] Compound <![CDATA[IC 50 ]]> Compound <![CDATA[IC 50 ]]> Compound <![CDATA[IC 50 ]]> 1 A 2 A 3 A 4 A 5 A 6 A 7 A 8 A 9 A 10 B 11 A 12 A 13 A 14 C 15 B 16 A 17 A 18 A 19 A 20 C 21 A 22 B 23 A 24 A 25 A 26 A 27 A 28 A 29 A 30 A

[0154] Biological test example-2: Inhibition rate detection of mutant phosphorylated LRRK2 (S935) in transfected cells:

[0155] Cell transfection

[0156] HEK293T cells were seeded in 12-well plates with DMEM medium at 1×10^6 cells per well and placed in a 37°C, 5% carbon dioxide cell culture incubator overnight. After 24 hours of culture, the cells were replaced with OPTI-MEM medium, which is conducive to transfection. Each well of cells was transfected with 2.5ug of pCMV-LRRK2-G2019S plasmid, and the ratio of transfection reagent PEI to plasmid was 3:1. After fully mixing, it was allowed to stand for about 15 minutes, and the mixture was added dropwise to the 12-well plate. After 6-8 hours of transfection, the OPTI-MEM medium was removed and replaced with DMEM medium. It was placed in a cell culture incubator for about 24 hours, and then 2 microliters of the compound's DMSO storage solution or DMSO (volume ratio of 0.1%) was added, mixed, and continued to be cultured for 16-18 hours.

[0157] Protein extraction

[0158] Add 50 μL AlphaLISA 1x lysis buffer to each sample, lyse the cells on ice for 20 minutes, centrifuge at 12000 rpm at 4°C for 30 minutes, take the supernatant, measure the concentration of the cell lysate by BCA, and normalize the concentrations of all samples.

[0159] Determination of pS935

[0160] Add 10 μL of sample of the same concentration to a 384-well plate, then add 5 μL of Acceptor Mix, seal the plate with a sealing film and cover with tin foil, mix well, incubate at room temperature for 2 hours, then add 5 μL of Donor Mix, mix well, incubate in the dark for 16-18 hours, and analyze the data after reading with a PE microplate reader.

[0161] Table 3 below provides the inhibitory activity IC values ​​of some compounds on LRRK2 (S935) in cells: 50 Range:

[0162] (IC 50 Range: A: <0.1μM; B: 0.1-0.5μM; C: >0.5μM)

[0163] Table 3 IC of the inhibitory activity of the compounds against LRRK2 (S935) in cells 50

[0164] Compound <![CDATA[IC 50 ]]> Compound <![CDATA[IC 50 ]]> Compound <![CDATA[IC 50 ]]> 1 A 2 B 3 A 4 A 5 A 6 A 7 A 8 A 9 A 10 C 11 A 12 A 13 C 14 C 15 C 16 A 17 A 18 A 19 A 20 C 21 A 22 B 23 A 24 A 25 A 26 C 27 A 28 A 29 C 30 C

[0165] Biological test example-3: Oral blood drug concentration test and brain permeability test in mice

[0166] The C57BL / 6JNifdc mice used in this experiment were provided by Zhejiang Weitong Lihua Experimental Animal Technology Co., Ltd. The compound was dissolved in DMSO / Solutol / Saline (v / v / v=10 / 10 / 80) as a stock solution (3 mg / mL). The stock solution was orally administered to 3 C57BL / 6JNifdc mice at a dose of 20 mg / kg. Blood was collected from the orbital venous plexus at 0.5 hours and 1 hour after oral administration. The mice were euthanized after 1 hour of blood collection, and the whole brain tissue was collected.

[0167] Plasma processing:

[0168] About 200 μL of blood sample was collected into EDTAK2 blood collection tube, and then immediately centrifuged at 10,000 rpm for 5 minutes. 66 μL of supernatant plasma was taken, 198 μL of acetonitrile (from Anage) was added and vortexed for 5 minutes, and then immediately centrifuged at 13,000 rpm for 8 minutes. 200 μL of supernatant was taken to detect blood drug concentration.

[0169] Brain tissue processing:

[0170] The collected whole brain tissue samples were weighed and placed in a pre-cooled grinding tube, 1 mL of acetonitrile was added, and the samples were ground evenly using a freezing grinder. The samples were then ultrasonicated for 20 minutes and centrifuged at 13,000 rpm for 20 minutes. 200 μL of the supernatant was taken and placed in a sample injection vial for testing.

[0171] Standard sample preparation:

[0172] Accurately weigh the drug and add acetonitrile to dissolve to 100 μg / mL. Use a pipette to draw 50 μL of the solution added to 4.95 mL of acetonitrile / water (v / v=3 / 1) to obtain a 1000 ng / mL mother solution. Take 650 μL of the mother solution and dilute the mother solution with acetonitrile / water (v / v=3 / 1) solution by 2 times gradient to obtain standard samples with concentrations of 500 ng / mL, 250 ng / mL, 125 ng / mL, 62.5 ng / mL, and 31.25 ng / mL, respectively.

[0173] Sample concentration monitoring:

[0174] The standard solutions with concentrations of 1000ng / mL, 500ng / mL, 250ng / mL, 125ng / mL, 62.5ng / mL, and 31.25ng / mL were detected by LC-MS (LC-MS2020 and LC-2030Plus of Shimadzu Corporation) in SIM mode to obtain the calibration curve. The treated blood samples were also detected by this method, and the calibration curve was used for quantitative analysis to obtain the drug concentration data of each treated blood sample.

[0175] Data processing:

[0176] Export the data and calculate the average blood drug concentration at each sampling time point:

[0177] Plasma drug concentration = drug concentration in treated blood sample × 4 / drug molecular weight; average blood drug concentration is the average of drug concentration in blood at each sampling time point.

[0178] Drug concentration in brain = drug concentration in treated brain tissue sample / weight of brain tissue; average blood drug concentration is the average of blood drug concentration at each sampling time point.

[0179] Table 4 below provides the drug concentrations in plasma and brain of some compounds after oral administration in mice:

[0180] Table 4 Drug concentrations of compounds when orally administered to mice

[0181]

[0182]

[0183] It can be seen from the above results that the compounds of the present invention have good oral plasma drug concentration and brain drug concentration, and can be used as oral drugs to treat neurodegenerative diseases.

[0184] The applicant declares that the present invention illustrates the sulfoximine compound and its application through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0185] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0186] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A sulfoximine compound or its stereoisomer, tautomer, isotope-labeled substance or pharmaceutically acceptable salt, characterized in that: The structure of the sulfoximine compound is shown in Formula I: Wherein, A is selected from hydrogen, substituted or unsubstituted any one of the following groups: C 1～6 Alkyl, C 3～8 Cycloalkyl, C 6～14 aryl, 5-12 membered heteroaryl, 3-8 membered heterocyclic group, 5-10 membered bridged ring group or 5-12 membered spirocyclic group, wherein the substituents substituted are selected from C 1～4 Alkyl, C 1～6 Alkoxy, C 3～6 Cycloalkyl, 3- to 8-membered heterocyclic group, C 1～3 Alkylphosphoryl, C 1～3 Alkylsulfonyl, oxo, cyano or halogen; L is selected from -H, -O-, -NH-, -S- or -N(C 1～4 alkyl)-, when L is -H, A does not exist; R1 is selected from hydrogen, cyano, hydroxyl, C 3～6 Cycloalkyl, substituted or unsubstituted C 1～6 Alkyl, substituted or unsubstituted C 1～6 Alkoxy, wherein the substituted substituent is selected from hydroxy, methylamino, dimethylamino, methoxy, carboxyl, oxo, cyano or halogen; R2 is selected from C 1～4 Alkyl, C 1～6 Alkoxy, C 3～6 Cycloalkyl, cyano or halogen; m is selected from 0, 1 or 2, when m is 0, the substituent is absent; when m is 2, the two R2 are the same or different; X is selected from CH or N; Y and Z are independently selected from C or N; R a is selected from hydrogen, cyano, halogen, C 1～3 Alkyl, C 1～3 Alkylamino, C 1～3 Alkylphosphoryl or C 1～3 Alkylsulfonyl; R b Selected from hydrogen, amino or C 1～3 Alkylamino; R a and R b It can exist independently or be linked together with the carbon atoms to form a 5- to 12-membered heteroaromatic ring or a 5- to 8-membered heterocyclic ring.

2. The sulfoximine compound or its stereoisomer, tautomer, isotope-labeled substance or pharmaceutically acceptable salt according to claim 1, characterized in that: The structure of the sulfoximine compound is shown in Formula II: Among them, A, L, R1, R2, m, X, Y, Z and R a It has the same definition as claim 1.

3. The sulfoximine compound or its stereoisomer, tautomer, isotope-labeled substance or pharmaceutically acceptable salt according to claim 1, characterized in that: The structure of the sulfoximine compound is shown in Formula III: wherein A, L, R1, R2, m, X, Y and Z have the same defined ranges as in claim 1; M is selected from C or N. When M is selected from C, R3 is selected from hydrogen, fluorine or chlorine; when M is selected from N, R3 is absent.

4. The sulfoximine compound or its stereoisomer, tautomer, isotope-labeled substance or pharmaceutically acceptable salt according to claim 1, characterized in that: The structure of the sulfoximine compound is shown in Formula IV: wherein A, L, R1, R2, m, X, Y and Z have the same defined ranges as in claim 1; R4, R5 are independently selected from hydrogen or fluorine, R6 is selected from hydrogen or C 1～3 alkyl.

5. The sulfoximine compound or its stereoisomer, tautomer, isotope-labeled substance or pharmaceutically acceptable salt according to any one of claims 1 to 4, characterized in that: The A is selected from the following structures: Preferably, said L is selected from -O- or -NH-.

6. The sulfoximine compound or its stereoisomer, tautomer, isotope-labeled substance or pharmaceutically acceptable salt according to any one of claims 1 to 4, characterized in that: The sulfoximine compound is selected from any one of the following structures:

7. Use of the sulfoximine compound or its stereoisomer, tautomer, isotope-labeled substance or pharmaceutically acceptable salt according to any one of claims 1 to 6 in the preparation of an LRRK2 protein kinase inhibitor.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises at least one sulfoximine compound according to any one of claims 1 to 6 or a stereoisomer, a tautomer, an isotope-labeled substance or a pharmaceutically acceptable salt thereof.

9. Use of the sulfoximine compound or its stereoisomer, tautomer, isotope-labeled substance or pharmaceutically acceptable salt according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 8 in the preparation of a drug for treating and / or preventing LRRK2 protein kinase-related diseases.

10. The use according to claim 9, characterized in that: The disease includes a neurodegenerative disease, an immune-related disease or a cancer, specifically including Parkinson's disease, Alzheimer's disease, Huntington's disease, Crohn's disease, inflammatory bowel disease, colitis, tuberculosis, leprosy, melanoma, non-melanoma skin cancer or breast cancer.

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