Pyridopyrrolic amino acid derivatives and their use in the preparation of leucine-rich repeat kinase 2 inhibitors

Abstract

The application discloses pyridopyrrole amino acid derivatives and application thereof in preparation of leucine-rich repeat kinase 2 inhibitors, which not only have good inhibition effect on LRRK2 activity, but also have structural characteristics of being actively transported by amino acid transporters on BBB, increase the ability of drugs into the central nervous system, and are potential therapeutic drugs for treating central nervous system degenerative diseases (such as Parkinson's disease).

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of chemical drugs, and provides a pyridopyrrole amino acid derivative and application thereof in preparation of leucine-rich repeat kinase 2 inhibitor. BACKGROUND

[0002] Parkinson's disease (PD) as a chronic neurodegenerative disease mainly affecting the elderly population, its incidence is second only to Alzheimer's disease. Since it was first discovered and named in 1817, the history of human struggle with it has been more than two centuries. According to the Parkinson's Disease Treatment Guidelines (Fourth Edition) of China, the prevalence of Parkinson's disease in people over 65 years old is about 1.7%, and more than 4% in people over 80 years old. It is estimated that by 2030, the number of Parkinson's disease patients in China will reach 5 million. At present, the awareness, treatment rate and diagnosis rate of Parkinson's disease are low, and there is no cure. Patients have to bear the burden of motor nervous system disorders such as tremor, limb rigidity, motor function decline and gait abnormalities, as well as non-motor symptoms such as hyposmia, sleep disorders, constipation and so on. The existing drug treatment can only relieve symptoms to a certain extent, but cannot stop the progression of the disease. The commonly used drugs in clinical practice have great deficiencies in meeting the needs of patients with Parkinson's disease in the middle and late stages, and there is an urgent need to develop a class of drugs that can prevent the pathological and biochemical regression of Parkinson's disease. Disease-modifying therapy is the mainstream direction of developing Parkinson's disease treatment drugs, which can affect the initial trigger factor of neuronal degeneration and promote neuronal compensation or reduce pathological transmission and progression. The current mainstream research believes that the aggregation of α-Syn in Lewy body (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.

[0003] And the leucine-rich repeat kinase 2 (Leucine-rich repeat kinase 2, LRRK2) blocks the chaperone-mediated autophagy, resulting in the inability of α-syn to be degraded and producing toxicity. LRRK2 is involved in α-syn-mediated neurotoxicity, and LRRK2 induces mitochondrial damage and endolysosomal dysfunction through an oxidative mechanism, triggering the progression of Parkinson's disease. LRRK2 kinase inhibitors can reduce pathological damage in Parkinson's disease models and improve motor dysfunction in patients. The safety and tolerability IB phase clinical trials of two new LRRK2 kinase inhibitors DNL201 and DNL151 were successful, and DNL151 has carried out IIb / III phase registered clinical research. Therefore, the development of LRRK2 small molecule inhibitors is one of the most potential research directions for developing Parkinson's disease treatment drugs.

[0004] Therefore, developing effective inhibitors of LRRK2 kinase and mutant LRRK2 kinase becomes an important way to treat neurodegenerative diseases. The present application aims to invent a compound that can highly inhibit LRRK2 kinase, so as to further invent a drug that can well treat neurodegenerative diseases.

[0005] Patent US8791112B2 discloses that pyrrolopyridine compounds of Arrien Company are a kind of LRRK2 inhibitors, and the chemical structure general formula is as follows.

[0006]

[0007] Although patent 148 has good LRRK2 inhibitory activity with IC50 activity <0.5 μM, it is found in experiments that the compound has low blood-brain barrier permeability and poor pharmacokinetic properties, and is stopped at the preclinical research stage. It is predicted that better blood-brain barrier permeable compounds need to be found for the target to ensure effective brain drug delivery while reducing drug concentration and side effects in peripheral tissues. SUMMARY

[0008] Therefore, the purpose of the present application is to provide a pyrrolopyridine amino acid derivative and its application in preparing leucine-rich repeat kinase 2 inhibitors.

[0009] To achieve the above purpose, the present application provides the following technical solutions.

[0010] 1. A pyrrolopyridine amino acid derivative or a pharmaceutically acceptable salt thereof, the derivative having a structure as shown in general formula (1):

[0011]

[0012] In general formula (1),

[0013] R1 is selected from H or C 1-5 alkyl;

[0014] R2 is selected from H or C 1-5 alkoxy.

[0015] Preferably, in general formula (1),

[0016] R1 is selected from H or methyl;

[0017] R2 is selected from H or methoxy.

[0018] Further preferably, in general formula (1),

[0019] R1 is H or methyl;

[0020] R2 is H.

[0021] Preferably, the derivative is (S)-2-amino-3-(3-(3-(cyclopropanecarbonyl)-lH- pyrrolo[2,3-b]pyridin-5-yl)phenyl)propionic acid ethyl ester, whose chemical structural formula is as follows:

[0022]

[0023] Preferably, the derivative is (S)-2-amino-3-(3-(3-(cyclopropanecarbonyl)-lH- pyrrolo[2,3-b]pyridin-5-yl)phenyl)propionic acid ethyl ester, whose chemical structural formula is as follows:

[0024]

[0025] Preferably, the derivative is (S)-2-amino-3-(3-(3-(cyclopropanecarbonyl)-lH- pyrrolo[2,3-b]pyridin-5-yl)phenyl)propionic acid ethyl ester, whose chemical structural formula is as follows:

[0026]

[0027] Preferably, the derivative is (S)-2-amino-3-(3-(3-(cyclopropanecarbonyl)-lH- pyrrolo[2,3-b]pyridin-5-yl)phenyl)propionic acid ethyl ester, whose chemical structural formula is as follows:

[0028]

[0029] Preferably, the derivative is (S)-2-amino-3-(3-(3-(cyclopropanecarbonyl)-lH- pyrrolo[2,3-b]pyridin-5-yl)phenyl)propionic acid ethyl ester, whose chemical structural formula is as follows:

[0030]

[0031] 2. Use of the aforementioned pyridopyrrolo amino acid derivative or a pharmaceutically acceptable salt thereof for the manufacture of a leucine-rich repeat kinase 2 inhibitor.

[0032] 3. Use of the aforementioned pyridopyrrolo amino acid derivative or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for preventing and / or treating a disease by inhibiting the activity of leucine-rich repeat kinase 2.

[0033] 4. Use of the aforementioned pyridopyrrolo amino acid derivative or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating or preventing a chronic neurodegenerative disease.

[0034] 5. Use of the aforementioned pyridopyrrolo amino acid derivative or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating or preventing Parkinson's disease.

[0035] 6. A pharmaceutical composition or preparation comprising the aforementioned pyrrolopyridine amino acid derivative or pharmaceutically acceptable salt thereof.

[0036] Preferably, it further comprises a pharmaceutically acceptable excipient, adjuvant or carrier.

[0037] The beneficial effects of the present application are:

[0038] The applicant introduces an amino acid fragment into the structure of the existing lead compound, aiming to maintain the LRRK2 inhibitory activity while increasing the active transport of the compound into the central nervous system through the amino acid transporters on the BBB, to find potential Parkinson's treatment drugs.

[0039] The present application provides a class of pyrrolopyridine structure amino acid derivatives, which not only have good inhibitory effect on LRRK2 activity, but also have the structural characteristics of being actively transported by amino acid transporters on the BBB, increasing the ability of the drug to enter the central nervous system, and is a potential treatment drug for treating central nervous system degenerative diseases (such as Parkinson's disease).

[0040] Other advantages, objects, and features of the present application will be apparent from the following specification and appended claims, and in part will be apparent to those skilled in the art in view of the foregoing disclosure, or can be learned by the practice of the present application. The advantages and features of the present application can be realized and obtained by means of the instrumentalities and combinations pointed out in the following specification. BRIEF DESCRIPTION OF DRAWINGS

[0041] In order to make the purposes, technical solutions and advantages of the present application clearer, the preferred detailed description of the present application will be combined with the drawings as follows, wherein:

[0042] Figure 1 The synthesis route of the target compound F1 of Example 1 is shown in the following scheme:

[0043] Figure 2 The synthesis route of the target compound F2 of Example 2 is shown in the following scheme:

[0044] Figure 3 The synthesis route of the target compound F3 of Example 3 is shown in the following scheme:

[0045] Figure 4 The synthesis route of the target compound F4 of Example 4 is shown in the following scheme:

[0046] Figure 5 The synthesis route of the target compound F5 of Example 5 is shown in the following scheme. DETAILED DESCRIPTION

[0047] The present application will be further described in combination with the specific embodiments.

[0048] DEFINITIONS AND DESCRIPTIONS

[0049] The following terms and phrases, as used herein, are intended to have the following meanings unless otherwise indicated. A particular term or phrase should not be construed as undefined or unclear without a specific definition, but should be interpreted according to its ordinary meaning. When a trade name appears herein, it is intended to denote the corresponding product or its active ingredient.

[0050] The term "pharmaceutically acceptable" as used herein pertains 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 problem or complication, commensurate with a reasonable benefit / risk ratio.

[0051] The term "pharmaceutically acceptable salt" means a salt of a compound of the present application that is within the scope of sound medical judgment, of a compound of the present application having particular substituents discovered in the present application, with a relatively nontoxic acid or base. Alkali addition salts can be prepared from the neutral forms of the compounds of the present application by contacting these compounds in pure solution or in a suitable inert solvent with a sufficient amount of the base to produce the neutral form of the compound. Pharmaceutically acceptable alkali addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts or similar salts. Acid addition salts can be prepared from the neutral forms of the compounds of the present application by contacting these compounds in pure solution or in a suitable inert solvent with a sufficient amount of the acid to produce the neutral form of the compound. Certain specific compounds of the present application contain both basic and acidic functionalities and can be converted into either alkali or acid addition salts.

[0052] The pharmaceutically acceptable salts of the present application can be synthesized from the parent compound that contains an acidic or basic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of the two.

[0053] The term "therapeutically effective amount" refers to the amount of a compound of the general formula that, when administered to a mammal in need of such treatment, is sufficient to effect treatment. The therapeutically effective amount will vary depending on the particular activity of the therapeutic agent employed, the age, physical condition, the presence of other disease states and nutritional status of the patient. In addition, the determination of the therapeutically effective amount of the therapeutic agent to be administered can be influenced by the other medications the patient can be receiving.

[0054] The term "treatment" means any treatment of a disease in a mammal, including: (i) preventing the disease, i.e., causing the clinical symptoms of the disease not to develop; (ii) inhibiting the disease, i.e., arresting the development of clinical symptoms; and / or (iii) relieving the disease, i.e., causing the regression of clinical symptoms.

[0055] 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. Such media and agents are those used in the art of pharmaceutical preparations, unless otherwise specified. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.

[0056] As used herein, the symbols and conventions used in these processes, schemes and examples are consistent with those used in the contemporary scientific literature, e.g., the Journal of the American Chemical Society or the Journal of Biological Chemistry. Unless otherwise indicated, all starting materials are obtained from commercial suppliers and are used without further purification. For example, the following abbreviations can be used throughout the examples and the specification: 1,4-Dioxane (1,4-dioxane), 4.0 M HC1 in Dioxane (4.0 M hydrochloric acid dioxane), 4.0 M HC1 in EA (4.0 M hydrochloric acid ethyl acetate), AcCl (acetyl chloride), ACN (acetonitrile), AcOK (potassium acetate, anhydrous), Ag2SO4(silver sulfate, anhydrous), AlCl3(aluminum trichloride), aq. (aqueous), (Boc)2O (di-tert-butyl dicarbonate), BPD (pinacol diboronic acid), DCM (dichloromethane), EA (ethyl acetate), FA (formic acid), g (grams), h (hours), HC1 (hydrochloric acid), Hz (hertz), I2(iodine), L (liters), L (microliters), LiOH (lithium hydroxide, anhydrous), m (milliliters), M (moles), mg (milligrams), MHz (megahertz), min (minutes), mM (millimoles), mmol (millimoles), mol (moles), Na2S2O3(sodium thiosulfate, anhydrous), NaHC03(sodium bicarbonate, anhydrous), Pd(dppf)Cl2([l,l-bis(diphenylphosphino) ferrocene] palladium dichloride), Pd(PPh3)4(tetrakis(triphenylphosphine) palladium); Na2CO3(sodium carbonate, anhydrous), r.t. (room temperature), TLC (thin layer chromatography).

[0057] The compounds of the present application can be prepared by a variety of synthetic processes known to those skilled in the art, including the specific embodiments set forth below, embodiments formed by the combination of the embodiments set forth with other chemical synthetic processes known to those skilled in the art, and equivalents thereof known to those skilled in the art, preferred embodiments including but not limited to the examples of the present application. Any intermediate or compound in a synthetic route that can be obtained by other reaction conditions is considered an alternative to the present application.

[0058] Example 1: Synthesis of target compound F1

[0059] The synthetic route is shown as Figure 1

[0060] The specific steps are as follows:

[0061] Step 1: Preparation of intermediate 3

[0062] First, AlCl3(54.1 g, 405.6 mmol) was dissolved in DCM (950 ml), then 5-bromo-7- azaindole (CAS: 183208-35-7, source: Shanghai Titan Science and Technology Co., Ltd., 10 g, 50.7 mmol) was added, stirred at room temperature 25℃ for 1 h, and finally cyclopropylcarbonyl chloride (CAS No.: 4023-34-1, source: Shanghai Titan Science and Technology Co., Ltd., 7.43 g, 71.0 mmol) was added, moved to an oil bath pot at 45℃ and reacted overnight. The reaction process was monitored by TLC plate. After the reaction was completed, the unreacted AlCl3 was filtered off with diatomite, and the filter cake was washed with DCM. The filtrate was poured into water and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (elution gradient: volume fraction of ethyl acetate / petroleum ether = 0-30%) to obtain intermediate 3 (8.44 g, 62%) as a white solid. [M+H] + = 265; [M+H] + = 267;

[0063] Step 2: Preparation of intermediate 4

[0064] Intermediate 3 (6 g, 22.63 mmol), BPD (5.75 g, 22.63 mmol), Pd(dppf)Cl2(828 mg, 1.13 mmol), AcOK (4.44 g, 45.26 mmol) were added to a 150 ml sealed tube, then 1,4-Dioxane (60 ml) was added, and the reaction was carried out in a nitrogen atmosphere at 110℃ oil bath for 2 h. The reaction process was monitored by TLC plate. After the reaction was completed, the reaction liquid was extracted with ethyl acetate, and the extracted organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (elution gradient: volume fraction of methanol / dichloromethane = 0-10%) to obtain intermediate 4 (2.61 g, 36%) as a white solid. [M+H] + = 313 (boron lipid); [M+H] + = 231 (boric acid).

[0065] Step 3: Preparation of intermediate 6​

[0066] (S)-3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid ethyl ester (CAS No.: 1159502-91-6, source: Shanghai Titan Science and Technology Co., Ltd., 230 mg, 0.61 mmol), intermediate 4 (231.45 mg, 0.74 mmol), Pd(PPh3)4(70.49 mg, 0.061 mmol), Na2CO3(129.31 mg, 1.22 mmol) were added to a 15 ml sealed tube, then 1,4-Dioxane (2 ml) / water (400 μl) was added, and the reaction was carried out under nitrogen protection in a 100 °C oil bath for 2 h. The reaction process was monitored by TLC plate. After the reaction was completed, the reaction liquid was extracted with ethyl acetate, and 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: volume fraction of methanol / dichloromethane = 0-10%) to obtain intermediate 6 (167 mg, 56.61%) as a white solid. [M+H] + = 478; [M-56] + = 422; [M-Boc] + = 378.

[0067] Step 4: Preparation of compound S9

[0068] Intermediate 6 (160 mg, 0.49 mmol) was added to a reaction test tube, then 4.0M HCl in EA solution (2 ml) was added, and the reaction was carried out at room temperature 25 °C for 2 h. The reaction process was monitored by TLC plate. After the reaction was completed, the crude product F1 was purified again by preparative chromatography column (elution gradient: volume fraction of ACN / 0.1% FA = 30%-70%), and then freeze-dried by a freeze dryer to obtain compound F1 (51.1 mg, 36.85%) as a white solid. [M+H] + = 378; HPLC = 95.01%; 1H NMR (600 MHz, DMSO-d6) δ 12.60 (s, 1H), 8.71 (s, 1H), 8.66 (d, J = 2.3 Hz, 1H), 8.59 (d, J = 2.2 Hz, 1H), 8.16 (s, 1H), 7.54 (dt, J = 7.9, 1.3 Hz, 1H), 7.51 (d, J = 1.9 Hz, 1H), 7.39 (t, J = 7.6 Hz, 1H), 7.20 (dt, J = 7.6, 1.4 Hz, 1H), 4.11 - 3.99 (m, 2H), 3.75 (t, J = 6.8 Hz, 1H), 2.95 (qd, J = 13.6, 6.8 Hz, 2H), 2.79 (tt, J = 7.9, 4.6 Hz, 1H), 1.09 (t, J = 7.1 Hz, 3H), 0.99 (p, J = 3.3 Hz, 2H), 0.91 (dt, J = 8.0, 3.4 Hz, 2H).

[0069] Example 2: Synthesis of target compound F2

[0070] The synthetic route is shown as Figure 2

[0071] The specific steps are as follows:

[0072] Step 1: Preparation of compound F2

[0073] Compound F1 (184 mg, 0.44 mmol) was added to a round-bottom flask, then ethanol (4 ml) / water (1 ml) was added, and finally lithium hydroxide (85.15 mg, 3.55 mmol) was added, and the reaction was carried out at room temperature 25°C for 2h, and the reaction process was monitored by TLC plate. After the reaction was completed, the pH of the reaction solution was adjusted to weakly acidic with aqueous formic acid (volume ratio of formic acid to water was 1:10); ethyl acetate was extracted, and the extracted organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The residual impurities in the crude product were filtered off by filter membrane, and the filtrate was evaporated to dryness to obtain the crude product. The crude product F2 was purified again by preparative chromatography column (elution gradient was ACN / 0.1% FA volume fraction = 30% to 70%), and compound F2 (71.6 mg, 46%) was obtained as a white solid by freeze-drying machine. [M+H] + = 350; HPLC = 98.85%; 1 ​H NMR (600 MHz, DMSO-d6) δ 8.72 - 8.67 (m, 2H), 8.64 - 8.59 (m, 1H), 7.59 (d, J = 1.9 Hz, 1H), 7.54 (dd, J = 8.4, 7.0 Hz, 1H), 7.39 (t, J = 7.6 Hz, 1H), 7.27 (d, J = 7.5 Hz, 1H), 3.44 (dd, J = 8.4, 4.3 Hz, 1H), 3.22 (dd, J = 14.3, 4.4 Hz, 1H), 2.93 (dd, J = 14.3, 8.2 Hz, 1H), 2.79 (ddd, J = 12.3, 7.9, 4.6 Hz, 1H), 1.05 - 0.96 (m, 2H), 0.90 (dq, J = 7.1, 3.3 Hz, 2H).

[0074] Example 3: Synthesis of F3

[0075] The synthetic route is shown as Figure 3

[0076] The specific steps are as follows:

[0077] Step 1: Preparation of intermediate 8

[0078] (S)-3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid ethyl ester (CAS: 591249-54-6, source: Shanghai Haohong Biomedical Technology Co., Ltd., 300 mg, 0.80 mmol), intermediate 4 (301.90 mg, 0.97 mmol), Pd(PPh3)4(93.12 mg, 0.08 mmol), Na2CO3(170.83 mg, 1.61 mmol) were added to a 15 ml sealed tube, then 1,4-Dioxane (5 ml) / water (1 ml) was added, and the reaction was carried out under nitrogen protection in a 100°C oil bath for 2 h. The reaction process was monitored by TLC plate. After the reaction was completed, the reaction liquid was extracted with ethyl acetate, and the extracted organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (elution gradient: volume fraction of methanol / dichloromethane = 0-10%) to obtain intermediate 8 (106.7 mg, 27.72%) as a white solid. [M+H] + = 478; [M-56] + = 422; [M-Boc] + = 378.

[0079] Step 2: Preparation of compound F3

[0080] ​Intermediate 8 (100 mg, 0.21 mmol) was added to the reaction tube, then 4.0 M HC1 in EA solution (2 ml) was added, and the reaction was allowed to proceed at room temperature 25 °C for 2 h, during which the reaction process was monitored by TLC plate. After the reaction was completed, the crude product was concentrated under reduced pressure. The crude product F3 was purified again by preparative chromatography column (elution gradient: volume fraction of ACN / 0.1% FA = 30% to 70%), and then freeze-dried by a freeze dryer to obtain compound F3 (18 mg, 20.93%) as a white solid. [M+H] + = 378; HPLC = 96.27%; 1 HNMR (600 MHz, DMSO-d6) δ 12.59 (s, 1H), 8.71 (s, 1H), 8.65 (d, J = 2.3 Hz, 1H), 8.59 (d, J = 2.2 Hz, 1H), 8.14 (s, 1H), 7.64 - 7.59 (m, 2H), 7.34 - 7.24 (m, 2H), 4.12 - 4.00 (m, 2H), 3.82 (t, J = 6.8 Hz, 1H), 3.00 - 2.91 (m, 2H), 2.79 (tt, J = 7.9, 4.6 Hz, 1H), 1.11 (t, J = 7.1 Hz, 3H), 1.01 - 0.94 (m, 2H), 0.90 (dq, J = 10.2, 3.3 Hz, 2H).

[0081] Example 4: Synthesis of target compound F4

[0082] Synthesis as shown in Figure 4 .

[0083] The specific steps are as follows:

[0084] Step 1: Preparation of compound F4

[0085] Compound F3 (150 mg, 0.36 mmol) was added to a round-bottom flask, followed by ethanol (4 ml) / water (1 ml), and then lithium hydroxide (69.44 mg, 2.89 mmol) was added, and the reaction was allowed to proceed at room temperature 25 °C for 2 h, during which the reaction process was monitored by TLC plate. After the reaction was completed, the reaction solution was adjusted to weakly acidic by using aqueous formic acid (volume ratio of formic acid to water = 1:10); ethyl acetate was used for extraction, and the extracted organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The residual impurities were filtered off by filter membrane, and the filtrate was evaporated to dryness to obtain the crude product. The crude product F4 was purified again by preparative chromatography column (elution gradient: volume fraction of ACN / 0.1% FA = 30% to 70%), and then freeze-dried by a freeze dryer to obtain compound F4 (43.1 mg, 34%) as a white solid. [M+H] + = 350; HPLC = 98.08%1 H NMR (600 MHz, DMSO-d6) δ 8.70 (s, 1H), 8.65 (d, J = 2.3 Hz, 1H), 8.58 (d, J = 2.3 Hz, 1H), 7.64 - 7.58 (m, 2H), 7.41 - 7.35 (m, 2H), 3.44 (dd, J = 8.2, 4.5 Hz, 1H), 3.18 (dd, J = 14.4, 4.6 Hz, 1H), 2.90 (dd, J = 14.3, 8.2 Hz, 1H), 2.78 (ddd, J = 12.5, 7.9, 4.6 Hz, 1H), 0.98 (p, J = 3.5 Hz, 2H), 0.90 (dt, J = 8.2, 3.5 Hz, 2H).

[0086] Example 5: Synthesis of F5

[0087] The synthetic route is shown as Figure 5

[0088] The specific steps are as follows:

[0089] Step 1: Preparation of intermediate 10

[0090] 4-methoxy-L-phenylalanine (CAS No.: 6230-11-1, source: Shanghai Titan Science and Technology Co., Ltd., 500 mg, 2.56 mmol) was dissolved in methanol (7 mL), AcCl (1 g, 12.8 mmol) was added dropwise to the reaction system at 0 °C, and after the addition was completed, it was continued to stir for 30 minutes, and then moved to an oil bath pot to heat and reflux for 4 hours. After the reaction was completed, the solvent was spun dry, and then ethanol (6 mL), NaHCO3(430 mg, 5.12 mmol), and (Boc)2O (559 mg, 2.56 mmol) were added, and after reaction at room temperature 25 °C for 5 hours, the reaction process was monitored by TLC plate. After the reaction was completed, the solvent was spun dry, poured into water with coarse filtration and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (elution gradient: volume fraction of ethyl acetate / petroleum ether = 0-10%), to obtain intermediate 10 (783 mg, 99%) as a colorless transparent liquid. [M+H] + = 310.

[0091] Step 2: Preparation of intermediate 11

[0092] ​Intermediate 10 (310 mg, 1.0 mmol) was dissolved in methanol (15 mL), at 0 °C, Ag2SO4(342 mg, 1.1 mmol), I2(280 mg, 1.1 mmol) were added to the reaction system, after stirring for 30 minutes, the reaction process was monitored by TLC plate. The reaction was completed, quenched with saturated Na2S2O3solution, and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (elution gradient: volume fraction of ethyl acetate / petroleum ether = 0-10%) to give intermediate 11 (431 mg, 90%) as a colorless transparent liquid. [M+H] + = 436.

[0093] Step 3: Preparation of intermediate 12

[0094] Intermediate 11 (340 mg, 0.78 mmol), intermediate 4 (390 mg, 1.25 mmol), Pd(dppf)Cl2(57 mg, 0.078 mmol), Na2CO3(207 mg, 1.95 mmol) were added to a 15 ml sealed tube, then 1,4-Dioxane (4 ml) / water (800 μl) was added, and the reaction was carried out under nitrogen protection in a 100 °C oil bath for 2 h. The reaction process was monitored by TLC plate. After the reaction was completed, the reaction liquid was extracted with ethyl acetate, and the extracted organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (elution gradient: volume fraction of ethyl acetate / petroleum ether = 0-20%) to give intermediate 12 as a white solid. [M+H] + = 494.

[0095] Step 4: Preparation of compound F5

[0096] Intermediate 12 (230 mg, 0.466 mmol) was dissolved in methanol / water (4 / 1 mL), then LiOH (22 mg, 0.932 mmol) was added to the reaction system, and stirred at room temperature 25 °C for 2 h. The reaction process was monitored by TLC plate. After the reaction was completed, the pH was adjusted to about 4-5 with 10% acetic acid aqueous solution, and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product which was used directly for the next step without purification.

[0097] The obtained crude product was added into a 10 mL vial, and 4.0 M HC1 in Dioxane solution (4 mL) was added, stirred at room temperature 25 °C for 2 hours, the reaction process was monitored by LC-Ms. After the reaction was completed, the solvent was dried and separated and purified by preparative chromatograph (elution gradient was ACN / 0.1% FA in volume fraction = 30% ~ 70%), and compound F5 (22 mg, 12.46%) was obtained as a white solid by lyophilizer freeze-drying. + = 380; HPLC = 95.01%; 1 H NMR (400 MHz, DMSO-d6) δ 12.68 (s, 1H), 8.68 (s, 1H), 8.57 (s, 1H), 8.40 (s, 1H), 7.25 (s, 2H), 7.02 (d, J = 8.6 Hz, 1H), 3.72 (s, 3H), 3.48 (dd, J = 7.8, 4.4 Hz, 1H), 3.14 (dd, J = 14.4, 4.3 Hz, 1H), 2.92 (dd, J = 14.3, 7.8 Hz, 1H), 2.82 - 2.72 (m, 1H), 0.98 (t, J = 3.9 Hz, 2H), 0.89 (dt, J = 7.1, 3.9 Hz, 2H).

[0098] Biological activity test

[0099] Protein binding experiment:

[0100] Reagent consumables:

[0101] LRRK2 G2019S enzyme (Thermo), substrate (LRRKtide) (Thermo), ATP (Thermo) TR-FRET diluent (Thermo), pLRRKtide antibody (Thermo), 384-well plate (PE) DMSO (Solebp)

[0102] Experimental process:

[0103] All test compounds (including positive control and test samples) were diluted to 1 mM with DMSO to obtain the corresponding test compound solution. 35 μL of positive compound (see Table 1 for structure, similar structure compound disclosed in patent US8791112B2 of Arrien company, and synthesized according to the literature), 35 μL of test compound solution, 35 μL of blank solution were added to a 384-well plate, the plate was centrifuged at 2500 rpm for 1 minute, 10 points were diluted with 3-fold gradient, and the initial concentration was 1 mM. According to 100 nL of positive compound, test compound, and blank well solution per well, they were added to another 384 test plate, 3 replicates, the plate was centrifuged at 2500 rpm for 1 minute and sealed in the foil for use.

[0104] Enzyme reaction: Dilute LRRKtide substrate and LRRK2 G2019S kinase mix working solution (final concentration of LRRKtide substrate: 400 nM and LRRK2 G2019S kinase: 580 ng / mL) with assay buffer (Thermo TR-FRET Dilution buffer) and add 5 μL per well to all sample wells of the above 384 assay plate, and incubate the 384 assay plate at 23℃ for 20 minutes. After incubation, dilute 2x ATP working solution (134 μM) with assay buffer and add 5 μL per well, and incubate the 384 assay plate at 23℃ for 60 minutes.

[0105] Detection: Dilute EDTA and pLRRKtide antibody with assay buffer (TR-FRET Dilution buffer) to obtain a mixed working solution (final concentration of EDTA: 10 mM, pLRRKtide antibody: 2 nM). Then add 10 μL of the antibody mixed working solution to each well of the above 384 assay plate, and incubate at 23℃ for 60 minutes. Read the plate in TE-FRET mode with excitation at 340 nm and fluorescence emission at 520 nm and terbium emission at 490 nm.

[0106] Method reference: J De Bente Fidalgo et al. Compounds, compositions and methods: CN113939294A[P]. 2022-01-14.

[0107] The activity data of each compound is shown in Table 1.

[0108] Table 1. Compound activity data table

[0109]

[0110]

[0111] As can be seen from Table 1, the series of new compounds provided by the embodiments of the present application have strong inhibitory effect on the kinase LRRK2 G2019S. Compared with the inhibitory activity of the positive structure on LRRK2 G2019S, especially the compounds F2 provided in Example 2 and the compound F4 provided in Example 4, all show better inhibitory effect on the mutant kinase LRRK2 G2019S. The compound F1 provided in Example 1 and the compound F3 provided in Example 3 also have better inhibitory effect on the mutant kinase LRRK2 G2019S. The compound F5 provided in Example 5 shows comparable inhibitory effect on the mutant kinase LRRK2 G2019S as the positive structure. It has potential application value in the preparation of drugs for preventing and / or treating diseases related to the increase of LRRK2 activity in vivo.

[0112] Finally, it is to be explained that the above embodiments are only used to illustrate the technical solutions of the present application but not to limit the present application. Although the present application is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present application can be modified or equivalently replaced without departing from the purpose and scope of the technical solutions, and all should be covered in the scope of the claims of the present application.

Claims

1. A pyrrolopyridine amino acid derivative or a pharmaceutically acceptable salt thereof, characterized by, The structure of the derivative is shown in general formula (1): General formula (1) In general formula (1), R1is selected from H or C 1-5 alkyl; R2is selected from H or C 1-5 alkoxy.

2. The pyrrolopyridine amino acid derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized by In general formula (1), R1 is selected from H or methyl; R2 is selected from H or methoxy.

3. The pyrrolopyridine amino acid derivative or a pharmaceutically acceptable salt thereof according to claim 2, characterized by In general formula (1), R1 is H or methyl; R2 is H.

4. Use of the pyrrolopyridine amino acid derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 for the manufacture of a medicament for the treatment or prevention of Parkinson's disease.

5. A pharmaceutical composition or formulation, characterized in that, A pharmaceutical composition comprising the pyrrolopyridine amino acid derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3.

6. The pharmaceutical composition or formulation according to claim 5, wherein Further comprising a pharmaceutically acceptable adjuvant.

7. The pharmaceutical composition or formulation according to claim 5, wherein Further comprising a pharmaceutically acceptable carrier.

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