Pyrimidine amino aryl alanine derivative and application thereof as leucine-rich repeat kinase 2 inhibitor

Abstract

The invention discloses a pyrimidine amino aryl alanine derivative and application thereof as a leucine-rich repeat kinase 2 inhibitor, the derivative not only has a good inhibition effect on the activity of LRRK2, but also has the structural characteristic of active transport by amino acid transport protein on BBB, the ability of drugs to enter the center is increased, and the drug activity is improved. The compound is a potential therapeutic drug for treating central nervous degenerative diseases (such as Parkinson's disease).

Description

[0001] This application is a divisional application. The filing date of the original application is December 20, 2022, the application number is 2022116437484, and the name of the invention is: Pyrimidineaminoarylalanine derivatives and their use as leucine-rich repeat kinase 2 inhibitors. Technical Field

[0002] The invention belongs to the technical field of chemical drugs and relates to pyrimidine amino aryl alanine derivatives and applications thereof as leucine-rich repeat kinase 2 inhibitors. Background Art

[0003] Parkinson's disease (PD) is the second most common chronic neurodegenerative disease in the middle-aged and elderly, second only to Alzheimer's disease. Since it was discovered and named in 1817, humans have been fighting against it for more than 200 years. At present, people have low awareness of the disease, low consultation rate, low diagnosis rate, and it is incurable. Patients show lifelong motor nervous system disorders such as tremor, limb stiffness, motor dysfunction and gait abnormalities, as well as non-motor symptoms such as hyposmia, sleep disorders, and constipation. Existing drugs can only relieve the symptoms themselves to varying degrees and cannot control the progression of the disease. The current commonly used clinical drugs are far from meeting the needs of existing patients with Parkinson's disease in the middle and late stages. There is an urgent need for a class of drugs that can prevent Parkinson's pathology and biochemical degeneration. Disease-modifying therapy is the mainstream direction of the development of drugs for the treatment of Parkinson's disease. It can affect the initial triggering factors of neuronal degeneration and promote neuronal compensatory reactions or reduce pathological propagation and progression. Current mainstream research believes that the aggregation of α-Syn in Lewy bodies (LB) is an important cause of Parkinson's disease, and reducing the aggregation of α-Syn is a potential method for treating Parkinson's disease.

[0004] Leucine-rich repeat kinase 2 (LRRK2) blocks chaperone-mediated autophagy, resulting in the inability to degrade α-syn and causing toxicity. LRRK2 is involved in α-syn-mediated neurotoxicity. LRRK2 induces mitochondrial damage and endolysosomal dysfunction through oxidative mechanisms, inducing the progression of Parkinson's disease. LRRK2 kinase inhibitors can reduce pathological damage in Parkinson's disease models and improve patients' motor dysfunction. The safety and tolerability of two new LRRK2 kinase inhibitors, DNL201 and DNL151, were successfully tested in Phase IB clinical trials, and DNL151 has been launched into Phase IIb / III registration clinical studies. Therefore, the development of LRRK2 small molecule inhibitors is one of the most promising research directions for the development of Parkinson's treatment drugs.

[0005] Therefore, developing effective inhibitors of LRRK2 kinase and mutant LRRK2 kinase has become an important approach to treat neurodegenerative diseases. The present invention aims to invent a compound that can highly inhibit LRRK2 kinase, thereby further inventing a drug that can effectively treat neurodegenerative diseases.

[0006] Patent US8802674B2 discloses that Gentech's pyrimidine aminobenzamide compounds are a class of LRRK2 inhibitors, and their chemical structure is shown below. And J.Med.Chem.2012, 55, 9416-9433 discloses Gentech's preferred compound GNE-7915 of this general structure, and its chemical structure is shown below,

[0007]

[0008] Although GNE-7915 has good LRRK2 inhibitory activity and a certain blood-brain barrier penetration ability, animal experiments still found that it had obvious toxic side effects on peripheral tissues (such as kidneys and lungs) and stopped at Phase 1 clinical development, indicating that it is necessary to find better blood-brain barrier permeability compounds for this target in order to ensure effective drug delivery to the brain while reducing drug concentrations and toxic side effects in peripheral tissues.

[0009] The blood-brain barrier (BBB) ​​is composed of brain microvascular endothelial cells, endothelial cell tight junctions, glial cells, astrocytes, pericytes, basement membranes and other components. It usually only allows gas molecules and lipid-soluble molecules with a small relative molecular mass to pass through, so that most drugs for treating central nervous system diseases cannot cross the BBB to reach effective therapeutic concentrations in the brain. Another important feature of the BBB is that it contains a variety of substance transporters on the membrane of brain capillary endothelial cells, which are related to the transmembrane transport of brain nutrients. Amino acid transporters are one of them. Among the main members, L-type amino acid transporters (LATs) are closely related to drug transport. Among them, the most studied LAT1 can transport amino acid central nervous system drugs with similar substrate structures, such as the anti-Parkinson's drug levodopa. The structural characteristics of dopamine limit its blood-brain barrier penetration rate, while levodopa modified by amino acid structure is a LAT1 substrate, which can be transported through the blood-brain barrier by LAT1 and enter the central nervous system to exert therapeutic effects. Gabapentin, a drug used to treat neuralgia and epilepsy, also has an amino acid structure that can be transported into brain tissue through LAT1. Summary of the invention

[0010] In view of this, the object of the present invention is to provide a pyrimidine amino aryl alanine derivative and its use as a leucine-rich repeat kinase 2 inhibitor.

[0011] In order to achieve the above object, the present invention provides the following technical solutions:

[0012] 1. A pyrimidine amino aryl alanine derivative or a pharmaceutically acceptable salt thereof, wherein the structure of the derivative is shown in the general formula (1):

[0013]

[0014] In the general formula (1),

[0015] R5 is selected from H, C 1-3 Alkyl, oxo C 1-3 Alkyl, halogenated C 1-3 alkyl;

[0016] X is selected from -C-, or -N-;

[0017] R6 is selected from H, C 1-3 Alkyl, oxo C 1-3 Alkyl, halogenated C 1-3 Alkyl and various common O protecting groups;

[0018] R7 is selected from H, C 1-3 Alkyl, oxo C 1-3 Alkyl, halogenated C 1-3 Alkyl and various common N protecting groups.

[0019] Preferably, the derivative is

[0020] (S)-2-amino-3-(4-((4-((2-hydroxyethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propanoic acid, (S)-2-amino-3-(4-((4-((2-hydroxyethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methox yphenyl)propanoic acid, its chemical structure is as follows:

[0021]

[0022] Preferably, the derivative is

[0023] (S)-2-amino-3-(5-((4-((2-hydroxyethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-4-methoxypyridin-2-yl)propanoic acid,

[0024] (S)-2-amino-3-(5-((4-((2-hydroxyethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-4-methox ypyridin-2-yl)propanoic acid, its chemical structure is as follows:

[0025]

[0026] 2. Use of the aforementioned pyrimidine aminoaryl alanine derivatives or pharmaceutically acceptable salts thereof in the preparation of leucine-rich repeat kinase 2 inhibitors.

[0027] 3. Use of the aforementioned pyrimidine aminoaryl alanine derivatives or pharmaceutically acceptable salts thereof in the preparation of drugs for inhibiting the activity of leucine-rich repeat kinase 2 to prevent and / or treat diseases.

[0028] 4. Use of the aforementioned pyrimidineaminoarylalanine derivatives or pharmaceutically acceptable salts thereof in the preparation of drugs for treating or preventing chronic neurodegenerative diseases.

[0029] 5. Use of the aforementioned pyrimidineaminoarylalanine derivatives or pharmaceutically acceptable salts thereof in the preparation of drugs for treating or preventing Parkinson's disease.

[0030] 6. A pharmaceutical composition or preparation comprising the pyrimidineaminoarylalanine derivative or a pharmaceutically acceptable salt thereof according to claim 1.

[0031] Preferably, it further comprises pharmaceutically acceptable excipients, adjuvants or carriers.

[0032] The beneficial effects of the present invention are:

[0033] The applicant introduced amino acid fragments into the structure of existing lead compounds, aiming to maintain the inhibitory activity on LRRK2 while increasing the property of active transport of the compounds into the central nervous system through amino acid transporters on the BBB, in order to find potential drugs for the treatment of Parkinson's disease.

[0034] The present invention provides a class of amino acid derivatives with pyrimidine aminoaryl structures, which not only have a good inhibitory effect on LRRK2 activity, but also have the structural characteristics of being actively transported by amino acid transporters on the BBB, thereby increasing the ability of drugs to enter the central nervous system. The derivatives are potential therapeutic drugs for treating central nervous system degenerative diseases (such as Parkinson's disease). BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to make the purpose, technical solution and beneficial effects of the present invention clearer, the present invention provides the following drawings for explanation.

[0036] Figure 1 The synthetic route of F001 in Example 1;

[0037] Figure 2 The synthetic route of F002 in Example 2;

[0038] Figure 3 This is the synthetic route of F003 in Example 3. DETAILED DESCRIPTION

[0039] The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0040] Definition and Description

[0041] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered to be uncertain or unclear in the absence of a special definition, but should be understood according to its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or its active ingredient.

[0042] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0043] The term "pharmaceutically acceptable salt" refers to salts of compounds of the present invention, prepared from compounds with specific substituents discovered by the present invention with relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Certain specific compounds of the present invention contain basic and acidic functional groups and can be converted into either base or acid addition salts.

[0044] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, these compounds in free acid or base form are reacted with a stoichiometric amount of an appropriate base or acid to prepare.

[0045] The term "therapeutically effective dose" refers to an amount of a compound of the formula sufficient to be effective for treatment when administered to a mammal in need of such treatment. The therapeutically effective amount will vary depending on the specific activity of the therapeutic agent used, the age, physiological condition, presence of other disease states, and nutritional status of the patient. In addition, other drug treatments that the patient may be receiving will affect the determination of the therapeutically effective amount of the therapeutic agent to be administered.

[0046] The term "treat" means any treatment of a disease in a mammal, including: (i) preventing the disease, i.e., causing clinical symptoms of the disease not to develop; (ii) inhibiting the disease, i.e., arresting the development of clinical symptoms; and / or (iii) palliating the disease, i.e., causing regression of clinical symptoms.

[0047] The term "pharmaceutically acceptable excipient, adjuvant or carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Unless any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic composition is contemplated. Supplementary active ingredients may also be incorporated into the composition.

[0048] The compounds of the present invention can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art, and preferred embodiments include but are not limited to the embodiments of the present invention. Any intermediate or compound 1 in the synthetic route that can be obtained by other reaction conditions is considered an alternative to the invention.

[0049] Example 1: Synthesis of F-001

[0050] Synthesis route such as Figure 1 shown.

[0051] The specific steps are as follows:

[0052] Step 1: Preparation of intermediate 2

[0053] 2,4-dichloro-5-trifluoromethylpyrimidine (5g, 23.04mmol) and triethylamine (4.66g, 46.08mmol) were added to methanol (50mL) and cooled to -80°C. After cooling, methylamine hydrochloride (1.56g, 23.04mmol) was added to the reaction system in batches, and the reaction was carried out at -80°C for 2h. TLC was used to monitor the progress of the reaction, and the temperature was raised to room temperature after the reaction was completed. The solvent was removed by concentration under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (elution gradient was petroleum ether / ethyl acetate = 10 / 1, volume ratio) to obtain intermediate 2 (1.34g, 27.5%, white solid), [M+H] + =212.

[0054] Step 2: Preparation of intermediate 4

[0055] Add intermediate 3 (10.0 g, 42.0 mmol) and 50 mL of methanol to a 250 mL single-mouth bottle, and stir at room temperature to dissolve. Then weigh sodium methoxide (2.5 g, 46.0 mmol) and dissolve in 50 mL of methanol (exothermic), add the sodium methoxide solution to the reaction solution of intermediate 3, wash with 30 mL of methanol and add to the reaction bottle, stir at room temperature for 1.5 hours, precipitate solid, TLC monitoring raw material unreacted, place the reaction bottle in a 45 ° C oil bath and stir for 1 hour, TLC monitoring raw material reaction is complete. The reaction solution is concentrated to dryness under reduced pressure and extracted with ethyl acetate. The extracted organic phase is washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. Petroleum ether is added to the concentrated residue and slurried for 2 hours. Filter and dry to obtain intermediate 4 (9.65 g, 91.8%, off-white solid).

[0056] Step 3: Preparation of intermediate 5

[0057] Take 120mL sealed tube, add intermediate 4 (4.0g, 16.0mmol), palladium acetate (150mg, 0.67mmol), N,N-dimethylformamide (DMF)) 12mL, triethylamine (TEA) (6.67mL, 48.0mmol) and methyl acrylate (2.88mL, 32.0mmol), after addition, heat to 100℃ sealed tube reaction for 18h. LC-MS and TLC monitoring, the reaction is complete. Filter through diatomaceous earth, and concentrate the filtrate to dryness under reduced pressure at 50℃. Add 200mL of water to the concentrated residue and extract with ethyl acetate (100mL×3). The extracted organic phase is washed with saturated NaCl, dried over anhydrous sodium sulfate, filtered and concentrated to obtain 4.9g of crude product. The crude product is purified by silica gel column chromatography (elution gradient is petroleum ether / ethyl acetate = 5 / 1, volume ratio) to obtain intermediate 5 (3.2g, 78.3%, yellow solid), [M+H] + =256.

[0058] Step 4: Preparation of intermediate 6

[0059] Add intermediate 5 (3.2 g, 12.5 mmol), 10% Pd / C (350 mg), 65 mL of tetrahydrofuran to a 250 mL single-mouth bottle, replace with hydrogen three times, and heat to 40 ° C for 72 h. LC-MS monitoring shows that the reaction is complete. Filter through diatomaceous earth, and concentrate the filtrate under reduced pressure to obtain a crude product. The crude product is purified by silica gel column chromatography (elution gradient is petroleum ether / ethyl acetate = 3 / 1, volume ratio) to obtain intermediate 6 (2.3 g, 80.7%, yellow solid), [M+H] + =228.

[0060] Step 5: Preparation of intermediate 7

[0061] Add intermediate 6 (2.3 g, 10.1 mmol) and 15 mL of methanol to the reaction tube, stir at room temperature to dissolve. Weigh lithium hydroxide (720 mg, 30.3 mmol) and dissolve in 15 mL of water, add to the above reaction tube, and react at room temperature. LC-MS and TLC monitoring, the reaction is complete, add 5% formic acid aqueous solution to adjust the pH to 2, the reaction solution is extracted with dichloromethane, 50 mL × 5), and a small amount of methanol is added each time. The extracted organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated to obtain intermediate 7 (2.1 g, 97.3%, gray solid), which is directly used in the next step. [M+H] + =214.

[0062] Step 6: Preparation of intermediate 8

[0063] Take 48mL sealed tube, add intermediate 7 (2.1g, 9.8mmol) and trifluoromethanesulfonic acid (10mL), seal the tube and react at 100℃ for 5h. TLC and LC-MS monitoring, the reaction is complete. Cool to room temperature in an ice-water bath, add saturated NaHCO3 to adjust the pH to neutral, and extract the reaction solution with dichloromethane 4 times. The extracted organic phase is washed with saturated NaCl, dried over anhydrous sodium sulfate, filtered and concentrated to obtain intermediate 8 (900mg, 46.8%, black solid), [M+H] + =196.

[0064] Step 7: Preparation of intermediate 9

[0065] Take a 100mL single-mouth bottle, add intermediate 8 (900mg, 4.6mmol) and 20mL methanol, and stir at room temperature to dissolve. Weigh NaBH4 (520mg, 13.8mmol) and slowly add it to the above reaction bottle. A large amount of gas is released. After the addition is completed, react at room temperature for 2h. LC-MS and TLC monitoring show that the reaction is complete. Add 5% formic acid aqueous solution to adjust the pH to 2, then adjust the pH to neutral with saturated NaHCO3, and extract the reaction mixture with EA (50mL×4). The extracted organic phase is washed with saturated NaCl, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product intermediate 9 (805mg, 88.5%, black oil), which is directly used in the next step. [M+H] + =198, [M+H-18] + =180.

[0066] Step 8: Preparation of intermediate 10

[0067] To the mixture of intermediate 9 (805 mg, 4.1 mmol) and toluene (20 mL), p-toluenesulfonic acid monohydrate (1.5 g, 8.16 mmol) was added. After addition, the temperature was raised to 100 °C for 2 h. The reaction was completed as monitored by LC-MS and TLC. 50 mL of EA was added to the reaction solution, and the pH was adjusted to neutral with saturated NaHCO3. The liquids were separated, and the aqueous phase was extracted with EA (50 mL × 3). The extracted organic layer was washed with saturated NaCl, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain intermediate 10 (460 mg, 62.8%, black oil), which was directly used in the next step. [M+H] + =180.

[0068] Step 9: Preparation of intermediate 11

[0069] Intermediate 10 (460 mg, 2.6 mmol) was added to a mixture of dioxane (10 mL) and water (6 mL) and stirred at room temperature. NaHCO3 (650 mg, 7.7 mmol) and CbzCl (benzyloxycarbonyl chloride, 550 μl, 3.8 mmol) were added to the above mixture in sequence, and the reaction was carried out at room temperature for 3 h. The reaction was completed by LC-MS and TLC monitoring. 20 mL of water was added to the reaction solution, and the product was extracted with EA (30 mL × 2). The extracted organic layer was washed with saturated NaCl, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 880 mg of crude product. The crude product was purified by silica gel column chromatography (elution gradient was petroleum ether / ethyl acetate = 10 / 1, volume ratio) and dried to obtain intermediate 11 (330 mg, 41.0%, yellow oil). [M+H] + =314, [M+H-44] + =270.

[0070] Step 10: Preparation of intermediate 12

[0071] The intermediate 11 (240 mg, 0.76 mmol) was dissolved in DCM (5 mL), and then m-CPBA (meta-chloroperbenzoic acid, 198 mg, 1.15 mmol) and NaHCO3 (193 mg, 2.3 mmol) were added in sequence. After addition, the mixture was reacted at room temperature for 18 h. After monitoring by LC-MS and TLC, 20 mL of DCM and 20 mL of water were added to the reaction solution. The separated DCM layer was washed with 20 mL of 10% sodium thiosulfate, then washed with saturated NaCl, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 420 mg of crude product. The crude product was purified by silica gel column chromatography (elution gradient was petroleum ether / ethyl acetate = 3 / 1, volume ratio) and dried to obtain the intermediate 12 (217 mg, 86.0%, foamy white solid). [M+H] + =330, [M+H-44] + =286.

[0072] Step 11: Preparation of intermediate 13

[0073] Intermediate 12 (200 mg, 0.6 mmol) and ZnI2 (775 mg, 2.4 mmol) were added to 5 mL of toluene and heated to 50 °C under N2 protection for 60 h. After the reaction was completed by LC-MS and TLC monitoring, EA was added to the reaction solution for extraction. The extracted organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (elution gradient: petroleum ether / ethyl acetate = 3 / 1, volume ratio) and dried to obtain intermediate 13 (50 mg, 25.0%, oil). [M+H] + =330, [M+H-44] + =286.

[0074] Step 12: Preparation of intermediate 14

[0075] Intermediate 13 (48 mg, 0.146 mmol) was dissolved in a mixture of ethanol (2 mL) and water (2 mL), followed by the addition of ammonium bicarbonate (230 mg, 2.91 mmol), cesium fluoride (67 mg, 0.44 mmol), and trimethylsilyl cyanide (55 ul, 0.44 mmol). After the addition was completed, the temperature was raised to 50 ° C for 24 h. The reaction was completed by LC-MS and TLC monitoring. 5 mL of water was added to the reaction solution, and the product was extracted with EA (5 mL × 3). The extracted organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 40 mg of crude product. The crude product was separated by preparative plate to obtain intermediate 14 (15 mg, 25.7%, oil). [M+H]+ =400, [M+H-44] + =356.

[0076] Step 13: Preparation of intermediate 15

[0077] Intermediate 14 (19 mg, 0.048 mmol) and palladium carbon (5 mg, palladium content 10%) were added to 2 mL of methanol, replaced with hydrogen three times, and reacted at room temperature for 2 h. LC-MS and TLC monitoring showed that the reaction was complete. The reaction solution was filtered and concentrated to obtain Intermediate 15 (17 mg, 134.7%, oil). [M+H] + =266.

[0078] Step 14: Preparation of target compound F-001

[0079] To a solution of intermediate 15 (9 mg, 0.034 mmol) and intermediate 2 (7.2 mg, 0.034 mmol) in tert-butanol (1.5 mL) was added glacial acetic acid (15 μL), and the temperature was raised to 90°C and refluxed for 20 h. The reaction was completed by LC-MS and TLC monitoring. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated by a preparative plate to obtain the target compound F-001 (6 mg, 40.1%, off-white solid). [M+H] + =441.1H NMR (400MHz, DMSO-d6) δ10.84(s,1H),8.48(s,1H),8.30(s,1H),8.20(s,1H),8.00(d,J=11.2Hz,1H),7.28(s,1H),3.76(s, 3H), 3.46 (d, J = 16.8Hz, 1H), 3.31 (d, J = 16.5Hz, 1H), 3.20 (d, J = 16.8Hz, 1H), 3.07 (d, J = 16.5Hz, 1H), 2.92 (d, J = 4.4Hz, 3H).

[0080] Example 2: Synthesis of target compound F-002

[0081] Synthesis Figure 2 shown.

[0082] The specific steps are as follows:

[0083] Step 1: Preparation of intermediate 2

[0084] 4-Bromo-2-methoxyaniline (8g, 39.80mmol) and sodium bicarbonate (8.69g, 103.48mmol) were added to a mixture of 1,4-dioxane (80mL) / water (48mL). After the addition was completed, the temperature was lowered to 0°C. Benzyl chloroformate (10.86g, 63.68mmol) was slowly added dropwise to the above reaction system. After the addition was completed, the reaction was allowed to react at room temperature for 0.5h. The reaction process was monitored by TLC plate. After the reaction was completed, the reaction solution was extracted with ethyl acetate, the extracted organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (elution gradient was petroleum ether / ethyl acetate = 3 / 1, volume ratio) to obtain intermediate 2 (12.6g, 94.7%, white solid). [M+H] + =336; [M+H] + =338; [M-44] + =292; [M-44] + =294.

[0085] Step 2: Preparation of intermediate 3

[0086] Under nitrogen protection, add Zn powder (9.37g, 143.28mmol) and dry DMF (10mL) to the first dried three-necked flask (100mL). After the addition, cool to 0℃. Weigh elemental iodine (4.55g, 17.91mmol) and dissolve it in ultra-dry DMF (2mL). Slowly add the iodine solution to the Zn powder solution with a syringe. After the addition, stir at 0℃ for 0.5h. Under nitrogen protection, add (R)-N-tert-butyloxycarbonyl-3-iodoalanine methyl ester (17.68g, 53.73mmol) and 10mL ultra-dry DMF to the second dried three-necked flask (100mL). Then draw out the mixed solution with a syringe and inject it into the first three-necked flask. React at room temperature for 2h. During the reaction, use a TLC plate to detect whether (R)-N-tert-butyloxycarbonyl-3-iodoalanine methyl ester has reacted. Under nitrogen protection, 2-dicyclohexylphosphine-2',6'-dimethoxybiphenyl (734.85 mg, 1.79 mmol), tris(dibenzylideneacetone)dipalladium (824.12 mg, 0.90 mmol), intermediate 2 (6 g, 17.91 mmol) and 20 mL of ultra-dry DMF were added to a third dried three-necked flask (100 mL) and heated to 60° C. The reaction solution in the first three-necked flask was slowly added dropwise to the third three-necked flask. After the addition was completed, the mixture was kept at 60° C. for 2 h.

[0087] After the reaction is completed, the reaction solution is placed in an ice-water bath, and water is slowly added to quench the Zn reagent. Filter to remove excess zinc powder, and extract the filtrate with ethyl acetate. The extracted organic phase is washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product is purified by silica gel column chromatography (elution gradient is dichloromethane / methanol = 10 / 1, volume ratio) to obtain intermediate 3 (5.2 g, 63.4%, light yellow oil). [M+H] + =459; [M-56] + =403; [M-Boc] + =359; [M-44] + =415.

[0088] Step 3: Preparation of intermediate 4

[0089] Add intermediate 3 (2.5 g, 5.36 mmol), 10% palladium carbon (5.7 g, 5.36 mmol) and tetrahydrofuran (25 mL) into a 100 mL round-bottom flask, replace the hydrogen and react at room temperature for 2 h. If the reaction is not complete after 2 h, add 0.1 equivalent of glacial acetic acid for catalysis. After the reaction is completed, the reaction solution is filtered and concentrated under reduced pressure to obtain intermediate 4 (1.5 g, 88.2%, light red oil). [M+H] + =325; [M-56] + =269; [M-Boc] + =225.

[0090] Step 4: Preparation of intermediate 7

[0091] 2,4-Dichloro-5-trifluoromethylpyrimidine (3 g, 13.83 mmol) and TEA (2.80 g, 27.66 mmol) were added to EtOH (30 mL) and cooled to -80 °C. Ethanolamine (844.74 mg, 13.83 mmol) was added to the above reaction solution, and the reaction was carried out at -80 °C for 2 h. TLC was used to monitor the reaction progress. After the reaction was completed, the temperature was raised to room temperature. The crude product was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (elution gradient was petroleum ether / ethyl acetate = 10 / 1, volume ratio) to obtain intermediate 7 (1.16 g, 34.8%, white solid), [M+H] + =242.

[0092] Step 5: Preparation of intermediate 8

[0093] Intermediate 4 (200 mg, 0.62 mmol), intermediate 7 (178.35 mg, 0.74 mmol) and acetic acid (18.62 mg, 0.31 mmol) were added to t-BuOH (2 mL), and the temperature was raised to 80 ° C under nitrogen protection for 16 h. After the reaction was completed, the reaction solution was extracted with ethyl acetate. The extracted organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (elution gradient was dichloromethane / methanol = 10 / 1, volume ratio) to obtain intermediate 8 (300 mg, 92.0%, brown oil). [M+H] + =530; [M-56] + =474; [M-Boc] + =430.

[0094] Step 6: Preparation of intermediate 9

[0095] Intermediate 8 (300 mg, 0.57 mmol) and lithium hydroxide (27.06 mg, 1.13 mmol) were added to a mixture of MeOH (4 mL) / H2O (1 mL). After addition, the mixture was reacted at room temperature for 2 h. After the reaction was completed, the pH of the reaction solution was adjusted to weak acidity using an aqueous formic acid solution (1:10), and then ethyl acetate was added for extraction. The extracted organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain intermediate 9 (252 mg, 86.30%, brown oil). [M+H] + =516; [M-56] + =460; [M-Boc] + =416.

[0096] Step 7: Preparation of target compound F-002

[0097] Intermediate 9 (250 mg, 0.49 mmol) was added to 3 mL of a 1,4-dioxane solution (3.0 M) of hydrochloric acid and reacted at room temperature for 2 h. After the reaction was completed, the crude product was filtered.

[0098] 1 mL of isopropanol was added to the crude product, and the temperature was raised to 80°C to dissolve the product. After the product was dissolved, it was transferred to room temperature and slowly stirred for 1 hour. The product precipitated, filtered, and dried to obtain the target compound F-002 (141.3 mg, 64.55%) as a white solid with a purity of 97.11%. [M+H] +=416.1H NMR(400MHz,DMSO-d6)δ9.85(s,1H),8.81–8.13(m,5H),7.86(s,1H),7.17(s,1H),6.88(dd ,J=8.1,1.7Hz,1H),4.24–4.12(m,1H),3.86(s,3H),3.61–3.46(m,4H),3.28–3.06(m,2H).

[0099] Example 3: Synthesis of target compound F-003

[0100] Synthesis route such as Figure 3 shown.

[0101] The specific steps are as follows:

[0102] Step 1: Preparation of intermediate 2

[0103] Under nitrogen protection, add Zn powder (1.57 g, 24 mmol) and ultra-dry DMF (10 mL) to the first three-necked flask (100 mL) that has been dried. After the addition is complete, cool to 0°C. Weigh elemental iodine (761 mg, 3 mmol) and dissolve it in ultra-dry DMF (5 mL). Slowly drop the iodine solution into the Zn powder solution. After the addition is complete, stir at 0°C for 0.5 h. Then add a DMF solution (5 mL, ultra-dry) of (R)-N-tert-butyloxycarbonyl-3-iodoalanine methyl ester (2.96 g, 9 mmol) to the above reaction system. After the addition is complete, react at room temperature for 2 h. During the reaction, use a TLC plate to detect whether the (R)-N-tert-butyloxycarbonyl-3-iodoalanine methyl ester has reacted.

[0104] Under nitrogen protection, bis(triphenylphosphine)palladium dichloride (60 mg, 0.15 mmol), intermediate 1 (566 mg, 3 mmol) and 10 mL of ultra-dry DMF were added to another three-necked flask (100 mL) and heated to 70° C. The organozinc reagent prepared in the first three-necked flask was slowly added dropwise to the mixed solution of intermediate 2 above, and the mixture was kept at 70° C. for 2 h.

[0105] After the reaction is completed, the reaction solution is placed in an ice-water bath, and a saturated ammonium chloride solution is slowly added to quench the zinc reagent. Filter to remove excess zinc powder, and extract the filtrate with ethyl acetate. The extracted organic phase is washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product is purified by silica gel column chromatography (elution gradient is petroleum ether / ethyl acetate = 3 / 1, volume ratio) to obtain intermediate 2 (496 mg, 46.3%, light yellow oil), [M+H] + =356.

[0106] Step 2: Preparation of intermediate 3

[0107] Intermediate 2 (480 mg, 1.35 mmol) was dissolved in 10 mL of anhydrous ethanol. After the solution was clear, iron powder (277 mg, 4.05 mmol), ammonium chloride (360 mg, 6.8 mmol) and 3 mL of water were added to the reaction solution. After the addition was complete, the temperature was raised to 50 ° C and stirred for 6 h. After the reaction was completed, the temperature was lowered to room temperature. After filtration, the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (elution gradient was petroleum ether / ethyl acetate = 3 / 1, volume ratio) to obtain intermediate 3 (356.9 mg, 81.2%, light yellow oil), [M+H] + =325.

[0108] Step 3: Preparation of intermediate 5

[0109] 2,4-Dichloro-5-trifluoromethylpyrimidine (239.5 mg, 1.1 mmol) and DIPEA (N,N-diisopropylethylamine, 237.8 mg, 1.84 mmol) were added to methanol (10 mL) and stirred at room temperature. Intermediate 3 (300 mg, 0.92 mmol) was added to the above reaction solution, and after addition, the temperature was raised to 50 ° C for 4 h. The reaction was monitored by TLC and the temperature was raised to room temperature. The crude product was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (elution gradient was petroleum ether / ethyl acetate = 3 / 1, volume ratio) to obtain intermediate 5 (167.9 mg, 36%, white solid), [M+H] + =506.

[0110] Step 4: Preparation of intermediate 7

[0111] Intermediate 5 (101 mg, 0.2 mmol), ethanolamine (50 mg, 0.8 mmol) and triethylamine (80 mg, 0.8 mmol) were added to acetonitrile (2 mL). After the addition was complete, the mixture was heated to 50 °C and reacted for 2 h. After the reaction was completed by TLC monitoring, the mixture was heated to room temperature. The crude product was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (elution gradient was petroleum ether / ethyl acetate = 1 / 1, volume ratio) to obtain intermediate 7 (71 mg, 67.0%, white solid), [M+H] + =531.

[0112] Step 5: Preparation of intermediate 8

[0113] Intermediate 7 (58 mg, 0.11 mmol) and lithium hydroxide (5.3 mg, 0.22 mmol) were added to a mixture of MeOH (0.5 mL) / H2O (2 mL). After addition, the mixture was reacted at room temperature for 2 h. After the reaction was completed, the pH of the reaction solution was adjusted to weak acidity using formic acid aqueous solution (1:10), and then ethyl acetate was added for extraction five times (the product has a high polarity and requires multiple extractions). The extracted organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain intermediate 8 (32.7 mg, 58.1%, white solid), [M+H] + =517.

[0114] Step 6: Preparation of target compound F-003

[0115] Add 2 mL of 3.0 M 1,4-dioxane hydrochloride solution to intermediate 8 (30 mg, 0.058 mmol) and react at room temperature for 2 h. After the reaction is complete, filter to obtain a crude product.

[0116] 0.5 mL of isopropanol was added to the crude product, and the temperature was raised to 80°C to dissolve the product. After the product was dissolved, the mixture was transferred to room temperature and slowly stirred for 1 h. The product precipitated, filtered, and dried to obtain the target compound F-003 (11 mg, 42.3%, white solid) with a purity of 95%, [M+H] + =417.1H NMR(400MHz,DMSO-d6)δ9.37–9.19(m,2H),9.12–8.42(m,2H),8.33(s,1H),7. 76(s,1H),7.47(s,1H),4.62(t,J=7.3Hz,1H),4.11(s,3H),3.70–3.46(m,6H).

[0117] Biological activity test

[0118] Protein binding assay:

[0119] Reagents and consumables:

[0120] LRRK2 G2019S enzyme (Thermo Fisher), substrate (LRRKtide) (Thermo Fisher), ATP (Thermo Fisher), TR-FRET diluent (Thermo Fisher), pLRRKtide antibody (Thermo Fisher), 384-well plate (PE) DMSO (Solarbio)

[0121] Experimental process:

[0122] All test compounds (including positive control and test samples) were diluted to 1 mM with DMSO to obtain corresponding test compound solutions. 35 μL of positive compound (structural formula is shown in Table 1, similar structural compounds disclosed by Genentech in the literature (Strada AA, Liu X, Baker-Glenn C, et.al. Discovery of highly potent, selective, and brain-penetrable leucine-rich repeat kinase 2 (LRRK2) small molecule inhibitors. J Med Chem. 2012 Nov 26; 55(22): 9416-33., and synthesized with reference to the literature) solution, 35 μL of test compound solution, and 35 μL of blank solution were added to a 384-well plate, and the plate was centrifuged at 2500 rpm for 1 minute. With 1 mM as the initial concentration, 3-fold gradient dilution was performed for 10 points, and 100 nL of positive compound, test compound, and blank well solution were added to another 384-well assay plate, three replicates, and the plate was centrifuged at 2500 rpm for 1 minute and sealed in foil for use.

[0123] Enzyme reaction: Dilute the LRRKtide substrate and LRRK2G2019S kinase mixed working solution (final concentration of LRRKtide substrate: 400nM and LRRK2 G2019S kinase: 580ng / mL) with assay buffer (Thermo Fisher TR-FRET Dilution buffer) and add it to all sample wells of the above 384 assay plate, 5μL per well, and incubate the 384 assay plate at 23°C for 20 minutes. After incubation, dilute the 2×ATP working solution (134μM) with assay buffer and add it to each well, 5mL per well, and incubate the 384 assay plate at 23°C for 60 minutes.

[0124] Detection: Dilute EDTA and pLRRKtide antibody with assay buffer (TR-FRET Dilution buffer) to obtain a mixed working solution (final concentration of EDTA: 10mM, pLRRKtide antibody: 2nM). Then add 10μL of the antibody mixed working solution to each well of the above 384 assay plate and incubate at 23°C for 60 minutes. Read the plate in the TE-FRET mode with 340nm excitation light, 520nm fluorescence emission light and 490nm terbium emission light.

[0125] Method reference: J. DeBicente Fidalgo et al. Compounds, compositions and methods: CN113939294A[P].2022-01-14.

[0126] The activity data of each compound are shown in Table 1.

[0127] Table 1. Compound activity data

[0128]

[0129] As shown in Table 1, the series of new compounds provided in the examples of the present invention have a strong inhibitory effect on the kinase LRRK2 G2019S. Compared with the inhibitory activity of the positive structure on LRRK2 G2019S, especially the compound F-001 provided in Example 1 of the present invention and the compound F-003 provided in Example 3 have comparable or better inhibitory effects on the mutant kinase LRRK2 G2019S. They have potential application value in the preparation of drugs for preventing and / or treating diseases related to increased gene LRRK2 activity in vivo.

[0130] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined by the claims of the present invention.

Claims

1. A pyrimidine amino aryl alanine derivative or a pharmaceutically acceptable salt thereof, characterized in that: The structure of the derivative is shown in the general formula (1): In the general formula (1), R5 is selected from H, C 1-3 Alkyl, oxo C 1-3 Alkyl, halogenated C 1-3 alkyl; X is selected from -C-, or -N-; R6 is selected from H, C 1-3 Alkyl, oxo C 1-3 Alkyl, halogenated C 1-3 Alkyl and various common O protecting groups; R7 is selected from H, C 1-3 Alkyl, oxo C 1-3 Alkyl, halogenated C 1-3 Alkyl and various common N protecting groups.

2. The pyrimidine amino aryl alanine derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The derivative is (S)-2-amino-3-(4-((4-((2-hydroxyethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methoxyphenyl)propanoic acid, and its chemical structure is as follows:

3. The pyrimidine amino aryl alanine derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The derivative is (S)-2-amino-3-(5-((4-((2-hydroxyethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-4-methoxypyridin-2-yl)propanoic acid, and its chemical structure is as follows:

4. Use of the pyrimidine aminoaryl alanine derivative or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a leucine-rich repeat kinase 2 inhibitor.

5. Use of the pyrimidine aminoaryl alanine derivatives or pharmaceutically acceptable salts thereof according to claim 1 in the preparation of drugs for inhibiting the activity of leucine-rich repeat kinase 2 to prevent and / or treat diseases.

6. Use of the pyrimidine amino aryl alanine derivatives or pharmaceutically acceptable salts thereof according to claim 1 in the preparation of drugs for treating or preventing chronic neurodegenerative diseases.

7. Use of the pyrimidine aminoaryl alanine derivatives or pharmaceutically acceptable salts thereof according to claim 1 in the preparation of drugs for treating or preventing Parkinson's disease.

8. A pharmaceutical composition or preparation comprising the pyrimidineaminoarylalanine derivative or a pharmaceutically acceptable salt thereof according to claim 1.

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