Pyridone compound as shown in formula (C) and preparation method and application thereof

Abstract

The invention belongs to the technical field of chemical drugs, and relates to a compound as shown in a general formula (C), or a racemate thereof, or an isomer thereof and a pharmaceutically acceptable salt thereof, as a PCSK9 inhibitor, and a method for treating various specific diseases or symptoms by using the PCSK9 inhibitor.

Description

Technical Field

[0001] The present invention belongs to the technical field of chemical drugs, and relates to a pyridone compound, or an isomer, or a racemate, or a pharmaceutically acceptable salt thereof, and a preparation method and application thereof. As a PCSK9 inhibitor, and a method for treating various specific diseases or conditions using the same. Background Art

[0002] Proprotein convertase subtilisin / kexin type 9 (PCSK9), also known as neuro-apoptosis-regulated convertase 1 (NARC-1), is a prohormone-proprotein convertase in the subtilisin (S8) family of serine proteases, which is expressed in cells with proliferation and differentiation capabilities, including hepatocytes, renal interstitial cells, ileal and colonic epithelial cells, and embryonic telencephalic neurons, etc. Studies have found that PCSK9 plays a role in the differentiation of hepatocytes and nerve cells. It can not only specifically act on cholesterol biosynthesis or uptake, but also circulating PCSK9 can directly bind to the low-density lipoprotein receptor (LDLR) on the surface of hepatocytes, and be phagocytosed by hepatocytes together with LDLR, promoting the degradation of LDLR in hepatocytes and hindering the recycling of LDLR, thereby increasing the content of LDL cholesterol (LDL-C) in plasma. The increase in LDL-C expression is closely related to human dyslipidemia and cardiovascular-related diseases.

[0003] Currently, research is being conducted on inhibiting the function of PCSK9 or inhibiting the production of PCSK9. For example, attempts have been reported to inhibit its function with monoclonal antibodies directed against PCSK9 and to inhibit the production of PCSK9 by RNA interference. However, for patients with cardiovascular diseases, effective small molecule inhibitors are needed to inhibit the function of PCSK9. Summary of the Invention

[0004] In view of the problems existing in the prior art, the present application provides a compound represented by the general formula (C), or an isomer, or a racemate, or a pharmaceutically acceptable salt thereof, and a preparation method and application thereof. As a PCSK9 inhibitor, and a method for treating various specific diseases or conditions using the same.

[0005] Specifically, the present invention is achieved through the following technical solutions:

[0006] A compound represented by the general formula (C), or an isomer, or a racemate, or a pharmaceutically acceptable salt thereof, includes:

[0007]

[0008] Wherein, ring A is selected from Among them, X and Y form a 5- to 7-membered saturated or unsaturated ring, and the 5- to 7-membered saturated or unsaturated ring contains 0, 1, or 2 heteroatoms selected from O, N, and S;

[0009] B is selected from

[0010] Q is selected from N or CR1, and R1 is selected from H or a halogen;

[0011] T1 is selected from N or CH;

[0012] Among them, R2 is selected from H, an alkyl group, or a halogen, and there is one or more R2;

[0013] R3 is selected from hydrogen or represents that the hydrogen on the A ring is further substituted by oxo, an alkyl group, a halogen, an alkoxy group, an alkylthio group, a haloalkyl group, a haloalkoxy group, a cycloalkyl group, a cycloalkylalkyl group, or an alkynyl group. There is one or more R3, or adjacent R3s form an alkoxy group

[0014] R4 is selected from hydrogen, a halogen, a hydroxyl group, an alkoxy group, a haloalkoxy group, a substituted or unsubstituted alkyl group, a cyano group, -C(O)-O-alkyl, a phenylalkoxy group, a carboxyl group, a hydroxymethyl group, or a cycloalkyl group. The substituent is selected from a hydroxyl group, an amide group, a halogen, or substituted, and U1, U2, and U4 are independently selected from CH or N, U3 is selected from CH2 or NH, and there is one or more R4;

[0015] And when the A ring is selected from then the B ring is not

[0016] As a preferred technical solution of the present invention, in the general formula (C), the alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, 1-ethylpropyl, 2-methylbutyl, tert-pentyl, 1,2-dimethylpropyl, isopentyl, neopentyl, n-hexyl, isohexyl, sec-hexyl, tert-hexyl, neohexyl, 2-methylpentyl, 1,2-dimethylbutyl, 1-ethylbutyl.

[0017] As a preferred technical solution of the present invention, in the general formula (C), the alkoxy group is selected from methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy.

[0018] As a preferred technical solution of the present invention, in the general formula (C), the cycloalkyl group is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

[0019] As a preferred technical solution of the present invention, in the general formula (C), the halogen is selected from fluorine, chlorine, bromine, and iodine.

[0020] As a preferred technical solution of the present invention, in general formula (C), Q is selected from N.

[0021] As a preferred technical solution of the present invention, in general formula (C), Q is selected from CH, and R2 is selected from F.

[0022] As a preferred technical solution of the present invention, in general formula (C), T1 is selected from N.

[0023] As a preferred technical solution of the present invention, in general formula (C), R3 is selected from hydrogen, methyl, methoxy, cyclopropyl, cyclopropylmethyl, fluorine, chlorine, oxo, CHF2-O-.

[0024] As a preferred technical solution of the present invention, in general formula (C), R4 is selected from hydrogen, methyl, hydroxyl, hydroxymethyl, cyano, F, Cl, Br, -O-CH2-benzene, -COOH, -COOCH2CH3, amide, formamide, CH3-C(O)-, ethynyl, trifluoromethyl, difluoromethoxy, methoxy.

[0025] As a preferred technical solution of the present invention, in general formula (C), ring A is selected from

[0026] Further, as a preferred technical solution of the present invention, in general formula (C), ring A substituted by R3 is selected from:

[0027] Further, as a preferred technical solution of the present invention, in general formula (C), ring B substituted by R4 is selected from:

[0028]

[0029] As a preferred technical solution of the present invention, the said compound, or its isomer, or its racemate, or its pharmaceutically acceptable salt, in general formula (C), is selected from: C1-C13, C15-C95.

[0030] The present invention further provides a pharmaceutical composition, which is characterized in that it contains a therapeutically effective amount of the said compound, or its isomer, or its racemate, or its pharmaceutically acceptable salt and a pharmaceutically acceptable carrier.

[0031] The present invention further provides the pharmaceutical use of the said compound, or its isomer, or its racemate, or its pharmaceutically acceptable salt, specifically, the use in the preparation of a drug for treating a disease, and the said disease is a PCSK9 inhibitor-related disease, specifically selected from diseases such as hypercholesterolemia.

[0032] For clarity, the general terms used in the description of the compounds are defined herein.

[0033] Unless otherwise indicated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered indefinite or unclear if it is not specifically defined, but should be understood in its ordinary meaning. When a trade name appears in this text, it is intended to refer to the corresponding product or its active ingredient. The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions, and / or dosage forms that are within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals, and without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0034] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared from the compounds with specific substituents found in the present invention and pharmaceutically acceptable acids or bases.

[0035] In addition to the salt form, prodrug forms of the compounds provided by the present invention also exist. The prodrugs of the compounds described herein are readily chemically transformed under physiological conditions to convert to the compounds of the present invention. In addition, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in the in vivo environment.

[0036] Certain compounds of the present invention can exist in unsolvated or solvated forms, including hydrate forms. Generally, the solvated forms are equivalent to the unsolvated forms and are all included within the scope of the present invention.

[0037] The compounds of the present invention can exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereoisomers, (D)-isomers, (L)-isomers, atropisomers, and their racemic mixtures and other mixtures, such as enantiomer- or diastereomer-enriched mixtures, all of which mixtures are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and their mixtures are included within the scope of the present invention.

[0038] Optically active (R)- and (S)-isomers, as well as D and L isomers, atropisomers, etc., can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are resolved by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is typically accomplished by using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate formation from an amine).

[0039] The atoms of the molecules of the compounds of the present invention are isotopes, and isotope derivatization can generally extend half-life, reduce clearance, stabilize metabolism, and increase in vivo activity. In addition, an embodiment is included in which at least one atom is replaced by an atom having the same atomic number (number of protons) and a different mass number (protons and neutrons). Examples of isotopes included in the compounds of the present invention include hydrogen atoms, carbon atoms, nitrogen atoms, oxygen atoms, phosphorus atoms, sulfur atoms, fluorine atoms, chlorine atoms, which respectively include 2 H. 3 H. 13 C. 14 C. 15 N. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 36 In particular, radioactive isotopes that emit radiation as they decay, such as 3 H or 14 C can be used for local anatomy testing of pharmaceutical preparations or compounds in vivo. Stable isotopes neither decay nor change with their amount nor are they radioactive, so they can be used safely. When the atoms constituting the molecules of the compounds of the present invention are isotopes, the isotopes can be converted according to general methods by replacing the reagents used in the synthesis with reagents containing the corresponding isotopes.

[0040] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compounds may be labeled with radioactive isotopes, such as deuterium ( 2 H), iodine-125( 125 I) or C-14( 14C). Transformations of all isotopic compositions of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

[0041] Further, one or more hydrogen atoms of the compounds of the present invention are replaced by the isotope deuterium ( 2 H). After the compounds of the present invention are deuterated, they have effects such as extended half-life, reduced clearance rate, metabolic stability, and enhanced in vivo activity.

[0042] The preparation methods of the said isotopic derivatives generally include: phase transfer catalysis methods. For example, the preferred deuteration method uses a phase transfer catalyst (for example, tetraalkylammonium salts, NBu4HSO4). Using a phase transfer catalyst to exchange the methylene protons of the diphenylmethane compound results in a higher deuterium incorporation than reduction with borohydride in the presence of an acid (for example, methanesulfonic acid) using a deuterosilane (for example, triethylsilane-d) or with a Lewis acid such as aluminum trichloride.

[0043] The term "pharmaceutically acceptable carrier" refers to any formulation carrier or medium that can deliver an effective amount of the active substance of the present invention, does not interfere with the biological activity of the active substance, and is non-toxic and side-effect-free to the host or patient. Representative carriers include water, oils, vegetables and minerals, paste bases, lotion bases, ointment bases, etc. These bases include suspending agents, thickening agents, transdermal promoters, etc. Their formulations are well-known to those skilled in the art of the cosmetics field or the topical drug field. For other information about carriers, reference can be made to Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the content of which is incorporated herein by reference.

[0044] The term "excipient" generally refers to the carrier, diluent, and / or medium required for formulating an effective pharmaceutical composition.

[0045] For a drug or a pharmacological active agent, the term "effective amount" or "therapeutically effective amount" refers to a sufficient amount of the drug or agent that is non-toxic but can achieve the desired effect. For the oral dosage forms in the present invention, the "effective amount" of an active substance in the composition refers to the amount required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount varies from person to person, depending on the age and general condition of the recipient, and also depends on the specific active substance. The appropriate effective amount in a particular case can be determined by those skilled in the art through routine tests.

[0046] The terms "active ingredient", "therapeutic agent", "active substance" or "active agent" refer to a chemical entity that can effectively treat a target disorder, disease or condition.

[0047] "Optional" or "optionally" means that the subsequently described event or condition may but does not necessarily occur, and this description includes both the case where the described event or condition occurs and the case where the described event or condition does not occur.

[0048] The compounds of the present invention can be prepared by various synthetic methods well-known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by their combination with other chemical synthesis methods, and the equivalent substitution methods well-known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples of the present invention. Specific embodiments

[0049] The present application will be further described in detail below in conjunction with examples, but the embodiments of the present application are not limited thereto.

[0050] Example C1

[0051] Synthesis of 6′-(((1S,3S)-3-((5-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3′-bipyridin]-2-one

[0052]

[0053] Step 1: Synthesis of 5-bromo-2-(difluoromethoxy)pyridine

[0054] At room temperature, 5-bromopyridin-2-ol (10.0 g, 57.5 mmol, 1.00 eq) was dissolved in DMF (200 mL), cesium carbonate (22.5 g, 69.0 mmol, 1.20 eq) was added, and the reaction was stirred at 25 °C for 1.5 hours under nitrogen protection. Then, sodium 2-chloro-2,2-difluoroacetate (26.3 g, 172 mmol, 3.00 eq) was added, and the temperature was raised to 100 °C and stirred for 1.5 hours.

[0055] LCMS showed that the target product MS (RT = 0.537 min, m / z = 223.9 [M+H] + )). The temperature was cooled to room temperature, 600 ml of water was added, and it was extracted with 600 ml * 5 of ethyl acetate. The organic phases were combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Silica gel was used for sample mixing, and column chromatography was used for separation and purification, and the product was eluted with pure petroleum ether. Colorless oily liquid 5-bromo-2-(difluoromethoxy)pyridine (3.01 g, 13.4 mmol, 23.38% yield) was obtained, LCMS (ESI) m / z = 223.9 [M+H] + .

[0056] Step 2: Synthesis of 6-(difluoromethoxy)pyridin-3-amine

[0057] At room temperature, 5-bromo-2-(difluoromethoxy)pyridine (3.00 g, 13.4 mmol, 1.00 eq), ammonia water (2.35 g, 20.1 mmol, 2.58 mL, 30% purity, 1.50 eq), copper(I) iodide (510 mg, 2.68 mmol, 0.2 eq), potassium carbonate (2.78 g, 20.09 mmol, 1.50 eq), and L-proline (616.76 mg, 5.36 mmol, 0.40 eq) were added to N-methylpyrrolidone (30 mL). The temperature was raised to 140 °C in a sealed flask, and the reaction was stirred for 12 hours.

[0058] TLC (petroleum ether / ethyl acetate = 5 / 1) showed that the raw materials had completely reacted. 20 ml of aqueous solution was added, and the mixture was extracted with ethyl acetate (30 ml × 4). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The sample was mixed with silica gel and separated by column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain the yellow oily product 6-(difluoromethoxy)pyridin-3-amine (1.3 g, 8.12 mmol, 60.62% yield).

[0059] Step 3: Synthesis of 5-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-amine

[0060] At 0 °C in an ice bath, 6-(difluoromethoxy)pyridin-3-amine (1.00 g, 6.25 mmol, 1.00 eq) and potassium thiocyanate (4.86 g, 50.0 mmol, 4.84 mL, 8.00 eq) were dissolved in glacial acetic acid (3.00 mL). A solution of bromine (2.99 g, 18.8 mmol, 965 μL, 3.00 eq) in glacial acetic acid (1.00 mL) was slowly added dropwise, keeping the temperature of the reaction solution below 0 °C. After the addition was complete, the temperature was allowed to rise to 20 °C naturally, and the reaction was stirred at 20 °C for 8 hours.

[0061] TLC (petroleum ether / ethyl acetate = 2 / 1) showed that the raw materials had completely reacted. 5 ml of water was added, and the temperature was raised to 85 °C. The mixture was filtered while hot. The filter cake was added with 1.5 ml of acetic acid, and the temperature was raised to 85 °C and filtered while hot. The two filtrates were combined, and the pH was adjusted to 8 with ammonia water in an ice-water bath. A large amount of yellow solid precipitated out. The mixture was filtered under reduced pressure, and the filter cake was dried to obtain the yellow target product 5-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-amine (0.9 g, 4.14 mmol, 66.35% yield).

[0062] Step 4: Synthesis of 2-chloro-5-(difluoromethoxy)thiazolo[5,4-b]pyridine

[0063] At room temperature, 5-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-amine (0.90 g, 4.14 mmol, 1.00 eq) was dissolved in acetonitrile (15 mL). Under nitrogen protection, isoamyl nitrite (728 mg, 6.22 mmol, 837 μL, 1.50 eq) and copper(II) chloride (669 mg, 4.97 mmol, 161 μL, 1.20 eq) were added, and the mixture was stirred at 25 °C for 3 hours.

[0064] TLC (petroleum ether / ethyl acetate = 2 / 1) showed that the raw material reaction was complete. 8 mL of saturated ammonium chloride solution was added and stirred for 10 minutes, then 25 mL of water was added for dilution. The mixture was extracted with 80 mL of ethyl acetate four times, and the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Silica gel was added for mixing, and the product was separated and purified by column chromatography (petroleum ether / ethyl acetate = 1 / 0 - 0 / 1). The white solid target product 2-chloro-5-(difluoromethoxy)thiazolo[5,4-b]pyridine (0.7 g, 2.96 mmol, 71.39% yield) was obtained. LCMS (ESI) m / z = 236.9 [M+H] + 。

[0065] Step 5: Synthesis of 6'-(((1S,3S)-3-((5-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one

[0066] At room temperature, 2-chloro-5-(difluoromethoxy)thiazolo[5,4-b]pyridine (200 mg, 845 μmol, 1 eq), 1-[6-[[[(1S,3S)-3-aminocyclopentyl]amino]-3-pyridinyl]pyridin-2-one hydrochloride (259 mg, 845 μmol, 1.00 eq, HCl) and triethylamine (257 mg, 2.54 mmol, 352 μL, 3.00 eq) were added to DMSO (10.0 mL). Under nitrogen protection, the temperature was raised to 60 °C and the reaction was carried out for 12 hours.

[0067] LCMS showed that the target product MS (RT = 0.423 min, m / z = 471.1 [M+H] + 30 mL of aqueous solution was added to the reaction solution to quench the reaction. The mixture was extracted with 60 mL of ethyl acetate four times, and the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Silica gel was added for mixing, and the product was separated and purified by column chromatography (petroleum ether / ethyl acetate = 1 / 0 - 1 / 1). The yellow solid product 6'-(((1S,3S)-3-((5-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one (124 mg, 264 μmol, 31.18% yield) was obtained.

[0068] LCMS (ESI) m / z = 471.1 [M+H] +

[0069] 1 H NMR (400 MHz, CDCl3): δ 8.41 (d, J = 6.6 Hz, 1H), 7.93 (d, J = 2.6 Hz, 1H), 7.84 - 7.77 (m, 1H), 7.64 - 7.56 (m, 1H), 7.47 (ddd, J = 2.1, 6.8, 9.1 Hz, 1H), 7.44 - 7.37 (m, 1H), 7.02 - 6.88 (m, 2H), 6.53 (d, J = 9.0 Hz, 1H), 6.44 (d, J = 9.1 Hz, 1H), 6.27 (dt, J = 1.2, 6.7 Hz, 1H), 4.47 - 4.26 (m, 2H), 2.26 - 2.11 (m, 2H), 1.96 (tq, J = 6.6, 13.7 Hz, 2H), 1.65 - 1.49 (m, 2H).

[0070] Example C2

[0071] Synthesis of 6′ - (((1S,3S)-3 - ((7 - (difluoromethoxy)-[1,2,4]triazolo[1,5 - a]pyridin - 2 - yl)amino)cyclopentyl)amino)-2H - [1,3′ - bipyridin]-2 - one

[0072]

[0073] Step 1: Synthesis of 2,7 - dibromo - [1,2,4]triazolo[1,5 - a]pyridine

[0074] (7 - Bromo - [1,2,4]triazolo[1,5 - a]pyridin - 2 - amine (2.00 g, 9.39 mmol, 1 eq), copper(II) bromide (3.15 g, 14.1 mmol, 1.5 eq) and tert - butyl nitrite (1.45 g, 14.1 mmol, 1.5 eq) were dissolved in acetonitrile (120 mL) and stirred at 80 °C for 3 h. The MS value of the product was monitored by LCMS (RT = 0.452 min). The reaction mixture was cooled to room temperature, then diluted with water (200 mL) and extracted with ethyl acetate (80 mL × 2). The combined organic phases were washed with saturated brine (200 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated to give 2,7 - dibromo - [1,2,4]triazolo[1,5 - a]pyridine (2.00 g, 7.22 mmol, 76.9% yield). It was used directly in the next step without purification. LCMS (ESI) m / z = 277.9 [M+1] +

[0075] Step 2: Synthesis of 2-bromo-[1,2,4]triazolo[1,5-a]pyridin-7-ol

[0076] Dissolve 2,7-dibromo-[1,2,4]triazolo[1,5-a]pyridine (2.00 g, 7.22 mmol, 1 eq), potassium hydroxide (1.2 g, 21.7 mmol, 3 eq), t-Bu Xphos (675 mg, 1.59 mmol, 0.22 eq) and Pd2(dba)3 (728 mg, 794 μmol, 0.11 eq) in 1,4-dioxane (20 mL) and water (5 mL), and stir at 100 °C for 4 h. The MS value of the product was monitored by LCMS (RT = 0.358 min). Cool the reaction solution to room temperature, then dilute the reaction solution with water (60 mL) and ethyl acetate (40 mL). Discard the separated organic phase. Adjust the pH of the aqueous phase to pH = 5 with dilute hydrochloric acid, and then extract the aqueous phase with ethyl acetate (60 mL × 2). The concentrated organic phase was washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 2-bromo-[1,2,4]triazolo[1,5-a]pyridin-7-ol (860 mg, 3.97 mmol, 54.9% yield, 98.7% purity). Without purification, it was directly used in the next step. LCMS (ESI) m / z = 215.9 [M+1] +

[0077] Step 3: Synthesis of 2-bromo-7-(difluoromethoxy)-[1,2,4]triazolo[1,5-a]pyridine

[0078] Dissolve 2-bromo-[1,2,4]triazolo[1,5-a]pyridin-7-ol (400 mg, 1.84 mmol, 1 eq) and cesium carbonate (721 mg, 2.21 mmol, 1.2 eq) in N,N-dimethylformamide (10 mL), then stir at 20 °C for 1 h. Then add sodium difluorochloroacetate (844 mg, 5.53 mmol, 3 eq) to the reaction solution, and then stir at 100 °C for 3 h. TLC (petroleum ether / ethyl acetate = 3 / 1) showed that the reaction was complete. Cool the reaction solution to room temperature, dilute the reaction solution with water (100 mL), and extract with ethyl acetate (20 mL × 2). The concentrated organic phase was washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by reverse-phase preparation (column: C18 150×30 mm; mobile phase: [water (FA)-ACN]; gradient: 30%-60% B over 7 min) to obtain 2-bromo-7-(difluoromethoxy)-[1,2,4]triazolo[1,5-a]pyridine (230 mg, 871 μmol, 47.2% yield, 100% purity). LCMS (ESI) m / z = 263.9 [M+1]+

[0079] Step 4: Synthesis of 6'-(((1S,3S)-3-((7-(Difluoromethoxy)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one

[0080] Dissolve 2-bromo-7-(difluoromethoxy)-[1,2,4]triazolo[1,5-a]pyridine (100 mg, 379 μmol, 1 eq), 6'-(((1S,3S)-3-aminocyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one (123 mg, 455 μmol, 1.2 eq), Xantphos (17.5 mg, 30.3 μmol, 0.08 eq), sodium phenoxide (66.0 mg, 568 μmol, 1.5 eq) and Pd2(dba)3 (13.9 mg, 15.2 μmol, 0.04 eq) in dioxane (4 mL), then stir at 135 °C for 40 minutes under microwave conditions. The MS value of the product was monitored by LCMS (RT = 0.391 min). Filter the reaction solution, concentrate the obtained filtrate to get the crude product, and purify the crude product by reverse-phase preparation (column: C18 150×30 mm; mobile phase: [water(FA)-ACN]; gradient: 12%-42% B over 7 min) to obtain 6'-(((1S,3S)-3-((7-(difluoromethoxy)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one (49.0 mg, 103 μmol, 27.1% yield, 95.1% purity).

[0081] LCMS (ESI) m / z = 454.2 [M+1] +

[0082] 1 H NMR: (400 MHz, DMSO-d6): δ 8.63 (d, J = 7.3 Hz, 1H), 7.92 (d, J = 2.5 Hz, 1H), 7.63 - 7.22 (m, 4H), 7.14 (d, J = 2.6 Hz, 1H), 6.91 (d, J = 6.9 Hz, 1H), 6.78 - 6.70 (m, 2H), 6.52 (d, J = 9.0 Hz, 1H), 6.44 (d, J = 8.9 Hz, 1H), 6.26 (dt, J = 1.3, 6.7 Hz, 1H), 4.38 - 4.26 (m, 1H), 4.15 (qd, J = 6.8, 13.4 Hz, 1H), 2.22 - 2.09 (m, 2H), 2.02 - 1.82 (m, 2H), 1.64 - 1.43 (m, 2H).

[0083] Example C3

[0084] Synthesis of 6′-(((1S,3S)-3-((6-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3′-bipyridin]-2-one

[0085]

[0086] Step 1: Synthesis of 3-bromo-5-(difluoromethoxy)pyridine

[0087] At room temperature, 3-bromo-5-hydroxypyridine (40.0 g, 230 mmol, 1.00 eq) was dissolved in DMF (800 mL).

[0088] Cs2CO3 (89.9 g, 276 mmol, 1.20 eq) was added, and the reaction was stirred at 25 °C for 1.5 hours under nitrogen protection. Then, sodium 2-chloro-2,2-difluoroacetate (105 g, 690 mmol, 3.00 eq) was added, and the temperature was raised to 100 °C and stirred for 1.5 hrs.

[0089] TLC (petroleum ether / ethyl acetate = 5 / 1) showed that the raw materials had completely reacted and the target product was formed. 60 ml of water was added, and the mixture was extracted with ethyl acetate (50 ml × 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was mixed with silica gel and purified by column chromatography (the product was eluted with pure petroleum ether). Yellow oily 3-bromo-5-(difluoromethoxy)pyridine (6.3 g, 28.1 mmol, 12.23% yield) was obtained.

[0090] Step 2: Synthesis of 5-(difluoromethoxy)pyridin-3-amine

[0091] At room temperature, 3-bromo-5-(difluoromethoxy)pyridine (5.00 g, 22.3 mmol, 1.00 eq), ammonia water (3.91 g, 33.5 mmol, 4.30 mL, 30% purity, 1.50 eq), CuI (850 mg, 4.46 mmol, 0.20 eq), K2CO3 (4.63 g, 33.48 mmol, 1.5 eq), and N,N′-bis(2-furfuryl)oxamide (2.22 g, 8.93 mmol, 0.4 eq) were added to NMP (10 mL). The reaction was carried out in a sealed vessel, the temperature was raised to 145 °C, and the reaction was stirred for 36 hours.

[0092] TLC (petroleum ether / ethyl acetate = 1 / 1) showed that the raw material reaction was complete and a new main spot was formed. 100 mL of water was added to the reaction solution to quench it, and it was extracted with ethyl acetate (100 ml * 4). The organic phases were combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. It was separated and purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1). The target product 5-(difluoromethoxy)pyridin-3-amine (3 g, 18.7 mmol, 83.94% yield) was obtained.

[0093] Step 3: Synthesis of 6-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-amine

[0094] Under an ice bath at 0 °C, 5-(difluoromethoxy)pyridin-3-amine (2.00 g, 12.5 mmol, 1.00 eq) and KSCN (9.71 g, 100 mmol, 9.68 mL, 8.00 eq) were dissolved in AcOH (6 mL), and a solution of Br2 (5.99 g, 37.5 mmol, 1.93 mL, 3.00 eq) in AcOH (2 mL) was slowly added dropwise, keeping the temperature of the reaction solution below 0 °C. After the addition was complete, the temperature was naturally raised to 20 °C and the reaction was stirred for 8 hrs.

[0095] TLC (petroleum ether / ethyl acetate = 2 / 1) showed that the raw material reaction was incomplete and a new spot of the target product was formed. 5 ml of water was added, the temperature was raised to 85 °C, and it was filtered while hot. The filter cake was added with 1.5 ml of acetic acid, the temperature was raised to 85 °C, and it was filtered while hot. The two filtrates were combined, and the pH was adjusted to 8 with ammonia water in an ice-water bath. A large amount of yellow solid precipitated out, and it was filtered under reduced pressure. The filter cake was dried. It was mixed with silica gel and separated and purified by column chromatography (petroleum ether / ethyl acetate = 2 / 1). The yellow solid target product 6-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-amine (460 mg, 2.12 mmol, 16.96% yield) was obtained. LCMS (ESI) m / z = 218.0 [M+H] +

[0096] Step 4: Synthesis of 2-chloro-6-(difluoromethoxy)thiazolo[5,4-b]pyridine

[0097] At room temperature, 6-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-amine (450 mg, 2.07 mmol, 1.00 eq) was dissolved in acetonitrile (10 mL), protected by nitrogen, and isoamyl nitrite (364 mg, 3.11 mmol, 418 μL, 1.50 eq) and CuCl2 (334 mg, 2.49 mmol, 1.20 eq) were added, and the reaction was stirred at 25 °C for 12 hours.

[0098] LCMS showed that the target product MS (RT = 0.524 min, m / z = 236.9 [M+H]+ ) 20 mL of saturated ammonium chloride solution was added and stirred for 10 minutes, then diluted with 100 mL of water. It was extracted with 150 mL×4 of ethyl acetate, and the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. It was separated and purified by column chromatography (petroleum ether / ethyl acetate = 5 / 1). The white solid target product 2-chloro-6-(difluoromethoxy)thiazolo[5,4-b]pyridine (275 mg, 1.16 mmol, 56.09% yield) was obtained. LCMS (ESI) m / z = 236.9 [M+H] +

[0099] Step 5: Synthesis of 6'-(((1S,3S)-3-((6-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one

[0100] At room temperature, 2-chloro-6-(difluoromethoxy)thiazolo[5,4-b]pyridine (100 mg, 423 μmol, 1.00 eq), 1-[6-[[[(1S,3S)-3-aminocyclopentyl]amino]-3-pyridinyl]pyridin-2-one (114 mg, 423 μmol, 1 eq), and TEA (128 mg, 1.27 mmol, 176 μL, 3.00 eq) were added to DMSO (2.00 mL), and the mixture was stirred at 70 °C for 6 hours.

[0101] LCMS showed that the target product had MS (RT = 0.417 min, m / z = 471.2 [M+H] + ) The temperature was lowered to room temperature, diluted with 10 mL of water, extracted with ethyl acetate 60 mL×4, the organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. It was separated and purified by column chromatography (petroleum ether / ethyl acetate = 0 / 1). The light yellow solid target product 6'-(((1S,3S)-3-((6-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one (198 mg, 42 μmol, 99.58% yield) was obtained

[0102] LCMS (ESI) m / z = 471.2 [M+H] +

[0103] 11H NMR (400 MHz, DMSO-d6): δ 8.67 (d, J = 6.8 Hz, 1H), 8.01 (d, J = 2.4 Hz, 1H), 7.93 (d, J = 2.6 Hz, 1H), 7.60 (dd, J = 1.6, 6.8 Hz, 1H), 7.56 (d, J = 2.5 Hz, 1H), 7.51 - 7.07 (m, 3H), 6.97 (d, J = 6.9 Hz, 1H), 6.53 (d, J = 8.9 Hz, 1H), 6.44 (d, J = 8.9 Hz, 1H), 6.27 (dt, J = 1.2, 6.7 Hz, 1H), 4.46 - 4.28 (m, 2H), 2.28 - 2.11 (m, 2H), 2.06 - 1.87 (m, 2H), 1.69 - 1.45 (m, 2H).

[0104] Example C4

[0105] Synthesis of 1-[6-[[[(1S,3S)-3-(5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl]amino]-3-pyridinyl]pyridin-2-one

[0106]

[0107] Step 1: 5,6,7,8-Tetrahydro-[1,2,4]triazolo[1,5-a]pyridin-2-amine

[0108] At 25 °C, [1,2,4]Triazolo[1,5-a]pyridin-2-amine (2.50 g, 18.6 mmol, 1.00 eq), wet palladium on carbon (0.40 g, 10% purity) and HCl (3.65 mL, 36.5% purity, 2.00 eq) were dissolved in ethanol (30 mL). Subsequently, the mixture was purged with hydrogen three times and then reacted under a hydrogen (50 psi) atmosphere for 72 hours.

[0109] LCMS showed the MS value of the detected product. After the reaction was completed, it was filtered and concentrated to obtain 5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyridin-2-amine (2.50 g, 97.1% yield). MS (ESI) m / z = 139.1 [M + 1] +

[0110] Step 2: 2-Iodo-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyridine

[0111] 5,6,7,8 - Tetrahydro - [1,2,4]triazolo[1,5 - a]pyridin - 2 - amine (0.20 g, 1.45 mmol, 1.00 eq), potassium iodide (601 mg, 3.62 mmol, 2.50 eq), p - toluenesulfonic acid (997 mg, 5.79 mmol, 4.00 eq) and sodium nitrite (200 mg, 2.89 mmol, 2.00 eq) were dissolved in acetonitrile (2 mL) and water (0.40 mL), and then the temperature was raised to 50 °C and the reaction was carried out for 2 hours.

[0112] TLC (methylene chloride:methanol = 20:1) showed that the raw material (R f = 0.24) was consumed completely.

[0113] The reaction solution was concentrated, then diluted with ethyl acetate (50 mL) and water (70 mL), separated, and the aqueous phase was extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 50:1 to 1:1) to obtain the product 2 - iodo - 5,6,7,8 - tetrahydro - [1,2,4]triazolo[1,5 - a]pyridine (250 mg, 69.4% yield).

[0114] Step 3: 1 - [6 - [[[(1S,3S) - 3 - (5,6,7,8 - tetrahydro - [1,2,4]triazolo[1,5 - a]pyridin - 2 - yl)amino]cyclopentyl]amino] - 3 - pyridinyl]pyridin - 2 - one

[0115] 2 - Iodo - 5,6,7,8 - tetrahydro - [1,2,4]triazolo[1,5 - a]pyridine (200 mg, 1.00 eq), 1 - [6 - [[[(1S,3S) - 3 - aminocyclopentyl]amino] - 3 - pyridinyl]pyridin - 2 - one (217 mg, 1.00 eq), sodium tert - butoxide (232 mg, 2.41 mmol, 3.00 eq) and t - BuXPhos Pd G3 (63.8 mg, 0.10 eq) were dissolved in dioxane (15 mL), purged with nitrogen three times, and then the temperature was raised to 90 °C and the reaction was carried out for 12 hours.

[0116] LCMS showed that the MS value of the product was detected.

[0117] The reaction solution was concentrated to obtain the crude product. The crude product was purified by reverse-phase preparation (column: C18 150×30 mm; mobile phase: [water(FA)-ACN]; gradient: 2%-32% B over 7 min), and the product 1-[6-[[[(1S,3S)-3-(5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl]amino]-3-pyridinyl]pyridin-2-one was obtained (45 mg, 13.80% yield, 96.4% purity).

[0118] LCMS(ESI) m / z = 392.2 [M+1] +

[0119] 1 1H NMR (400 MHz, CD3OD): δ 7.98 - 7.89 (m, 1H), 7.66 - 7.54 (m, 2H), 7.49 - 7.36 (m, 1H), 6.61 (dd, J = 5.2, 8.8 Hz, 2H), 6.47 (dt, J = 1.2, 6.8 Hz, 1H), 4.40 - 4.27 (m, 1H), 4.06 (quin, J = 6.4 Hz, 1H), 3.94 (t, J = 6.0 Hz, 2H), 2.74 (t, J = 6.4 Hz, 2H), 2.66 (s, 1H), 2.35 - 2.16 (m, 2H), 2.09 - 1.87 (m, 5H), 1.66 - 1.51 (m, 2H).

[0120] Example C5

[0121] Synthesis of 1-[6-[[[(1S,3S)-3-[(7-cyclopropyl-6,8-dihydro-5H-[1,2,4]triazolo[1,5-a]pyrazin-2-yl)amino]cyclopentyl]amino]-3-pyridinyl]pyridin-2-one

[0122]

[0123] Step 1: 2-bromo-[1,2,4]triazolo[1,5-a]pyrazine

[0124] At 0 °C, [1,2,4]triazolo[1,5-a]pyrazin-2-amine (7.00 g, 51.8 mmol, 1.00 eq) and hydrobromic acid (29.3 mL, 48% purity, 5.00 eq) were dissolved in acetic acid (40 mL). Subsequently, a solution of sodium nitrite (7.15 g, 2.00 eq) dissolved in water (40 mL) was added dropwise to the reaction solution. After the addition, the reaction was kept at a certain temperature for 2 hours.

[0125] LCMS showed the detected MS value of the product.

[0126] The reaction mixture was concentrated, then diluted with water (100 mL) and ethyl acetate (60 mL), and the aqueous phase was extracted with ethyl acetate (20 mL * 3). The concentrated organic phase was washed with saturated brine (30 mL * 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 20:1 to 1:1) to obtain the product 2-bromo-[1,2,4]triazolo[1,5-a]pyrazine (3.50 g, 33.9% yield). LCMS (ESI) m / z = 200.9 [M+1] +

[0127] Step 2: 2-Bromo-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine

[0128] At 20 degrees, 2-bromo-[1,2,4]triazolo[1,5-a]pyrazine (3.00 g, 1.00 eq) was dissolved in ethanol (20 mL), and then lithium borohydride (2.00 M, 30.2 mL, 4.00 eq) was added to the solution. After the addition, the reaction solution was heated to 50 degrees and reacted for 5 hours.

[0129] LCMS showed that the product was detected.

[0130] After the reaction, the reaction mixture was quenched with 1M hydrochloric acid (800 mL) and then washed with ethyl acetate (30 mL x 2). The organic phase was discarded. The aqueous phase was adjusted to pH 9 with saturated sodium carbonate solution and then extracted with dichloromethane (40 mL x 5). The organic phase from the second extraction was washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by reverse phase chromatography (column: Waters Xbridge 150 x 25 mm 5 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 1% to 30% over 9 min) to obtain the product, 2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine (1.60 g, 52.3% yield). MS (ESI) m / z = 203.0 [M+1]. +

[0131] Step 3: 2-Bromo-7-cyclopropyl-6,8-dihydro-5H-[1,2,4]triazolo[1,5-a]pyrazine

[0132] Dissolve 2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine (0.70 g, 3.45 mmol, 1.00 eq), cyclopropylboronic acid (4441 mg, 5.17 mmol, 1.50 eq), copper(II) acetate (1.25 g, 6.90 mmol, 2.00 eq) and N,N-diisopropylethylamine (891 mg, 1.20 mL, 2.00 eq) in dichloromethane (2 mL), and then react at 20 °C for 12 hours.

[0133] LCMS showed the MS of the detected product.

[0134] Dilute the reaction solution with dichloromethane (50 mL) and water (30 mL), and then extract the aqueous phase with dichloromethane (10 mL × 4). The concentrated organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 20:1 to 1:1) to obtain the product 2-bromo-7-cyclopropyl-6,8-dihydro-5H-[1,2,4]triazolo[1,5-a]pyrazine (160 mg, 19.09% yield). MS (ESI) m / z = 243.0 [M+1] +

[0135] Step 4: 1-[6-[[[(1S,3S)-3-[(7-cyclopropyl-6,8-dihydro-5H-[1,2,4]triazolo[1,5-a]pyrazin-2-yl)amino]cyclopentyl]amino]-3-pyridinyl]pyridin-2-one

[0136] Dissolve 2-bromo-7-cyclopropyl-6,8-dihydro-5H-[1,2,4]triazolo[1,5-a]pyrazine (140 mg, 1.00 eq), 1-[6-[[[(1S,3S)-3-aminocyclopentyl]amino]-3-pyridinyl]pyridin-2-one (156 mg, 1.00 eq), sodium tert-butoxide (166 mg, 3.00 eq) and t-BuXPhos Pd G3 (45.7 mg, 0.10 eq) in dioxane (12 mL), displace with nitrogen three times, and then heat to 90 °C and react for 12 hours.

[0137] LCMS showed the MS value of the detected product.

[0138] The reaction solution was concentrated to obtain the crude product, and the crude product was purified by preparative reverse-phase chromatography (column: C18 150×30 mm; mobile phase: [water(FA)-ACN]; gradient: 5%-35% B over 7 min and column: C18 150×30 mm; mobile phase: [water(FA)-ACN]; gradient: 5%-35% B over 7 min), to obtain the product 1-[6-[[[(1S,3S)-3-[(7-cyclopropyl-6,8-dihydro-5H-[1,2,4]triazolo[1,5-a]pyrazin-2-yl)amino]cyclopentyl]amino]-3-pyridinyl]pyridin-2-one (20 mg, 8.03% yield).

[0139] LCMS(ESI) m / z = 433.2 [M+1] +

[0140] 1 1H NMR (400 MHz, CDCl3): δ 8.03 (d, J = 2.4 Hz, 1H), 7.52 (dd, J = 2.4, 8.8 Hz, 1H), 7.39 (ddd, J = 2.4, 6.8, 9.2 Hz, 1H), 7.30 (dd, J = 2.0, 7.2 Hz, 1H), 6.65 (d, J = 9.2 Hz, 1H), 6.44 (d, J = 8.8 Hz, 1H), 6.23 (dt, J = 1.2, 6.8 Hz, 1H), 4.77 (br dd, J = 1.2, 5.6 Hz, 1H), 4.28 - 4.12 (m, 2H), 4.07 - 3.95 (m, 3H), 3.79 (s, 2H), 3.12 (t, J = 5.6 Hz, 2H), 2.39 - 2.23 (m, 2H), 2.11 - 1.97 (m, 2H), 1.95 - 1.86 (m, 1H), 0.62 - 0.55 (m, 2H), 0.54 - 0.48 (m, 2H).

[0141] Example C6

[0142] Synthesis of 1-[5-[[[(1S,3S)-3-(5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl]amino]pyrazin-2-yl]pyridin-2-one

[0143]

[0144] Step 1: 1-[6-[[[(1S,3S)-3-[(7-cyclopropyl-6,8-dihydro-5H-[1,2,4]triazolo[1,5-a]pyrazin-2-yl)amino]cyclopentyl]amino]-3-pyridinyl]pyridin-2-one

[0145] Dissolve 2-iodo-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyridine (300 mg, 1.00 eq), 1-[5-[[[(1S,3S)-3-aminocyclopentyl]amino]pyrazin-2-yl]pyridin-2-one (327 mg, 1.00 eq), sodium tert-butoxide (347 mg, 3.61 mmol, 3.00 eq) and t-BuXPhos Pd G3 (95.7 mg, 0.10 eq) in dioxane (10 mL), displace with nitrogen three times, then heat to 90 °C and react for 12 hours.

[0146] LCMS shows the MS value of the detected product.

[0147] Concentrate the reaction solution to obtain the crude product. The crude product is purified by reverse-phase preparation (column: Waters Xbridge 150*25mm*5um; mobile phase: [water(ammonia hydroxide v / v)-ACN]; gradient: 0%-25% B over 10 min) to obtain the product 1-[6-[[[(1S,3S)-3-[(7-cyclopropyl-6,8-dihydro-5H-[1,2,4]triazolo[1,5-a]pyrazin-2-yl)amino]cyclopentyl]amino]-3-pyridinyl]pyridin-2-one (30 mg, 6.35% yield).

[0148] LCMS(ESI) m / z = 391.1 [M-1] +

[0149] 1 1H NMR: (400 MHz, CDCl3): δ 7.76 (d, J = 1.2 Hz, 1H), 7.63 (dd, J = 2.0, 6.8 Hz, 1H), 7.38 (ddd, J = 2.0, 6.8, 9.2 Hz, 1H), 6.64 (d, J = 9.2 Hz, 1H), 6.32 - 6.22 (m, 1H), 4.93 (br d, J = 6.8 Hz, 1H), 4.41 - 4.29 (m, 1H), 4.24 - 4.12 (m, 1H), 4.04 (br d, J = 6.8 Hz, 1H), 3.97 (t, J = 6.0 Hz, 2H), 2.78 (t, J = 6.4 Hz, 2H), 2.42 - 2.25 (m, 2H), 2.12 - 1.99 (m, 4H), 1.96 - 1.87 (m, 2H), 1.60 - 1.51 (m, 2H).

[0150] Example C7

[0151] Synthesis of 6′-(((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl)amino)-2H-[1,3′-bipyridin]-2-one

[0152]

[0153] Step 1: Synthesis of 2-bromo-[1,2,4]triazolo[1,5-a]pyridine

[0154] Dissolve [1,2,4]triazolo[1,5-a]pyridin-2-amine (1.90 g, 14.2 mmol, 1.0 eq), copper(II) bromide (4.75 g, 21.3 mmol, 1.5 eq) and tert-butyl nitrite (2.19 g, 21.3 mmol, 1.50 eq) in acetonitrile (30 mL), and stir at 70 °C for 4 h. TLC (petroleum ether / ethyl acetate = 1 / 1) shows that the reaction is complete. Cool the reaction mixture to room temperature, then dilute it with water (50 mL) and extract with ethyl acetate (30 mL × 2). Combine the organic phases, wash with saturated brine (100 mL × 2), dry over anhydrous sodium sulfate, filter, and concentrate to obtain 2-bromo-[1,2,4]triazolo[1,5-a]pyridine (2.17 g, 11.0 mmol, 77.4% yield). Use it directly in the next step without purification. LCMS MS (ESI) m / z = 200.0 [M+1] + 。

[0155] Step 2: Synthesis of 6'-(((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl)amino)-2H-[1,3′-bipyridin]-2-one

[0156] 2-Bromo-[1,2,4]triazolo[1,5-a]pyridine (100 mg, 505 μmol, 1.0 eq), 6′-(((1S,3S)-3-aminocyclopentyl)amino)-2H-[1,3′-bipyridin]-2-one (150 mg, 556 μmol, 1.1 eq), Xantphos (23.4 mg, 40.4 μmol, 0.08 eq), sodium phenoxide (87.9 mg, 758 μmol, 1.5 eq) and Pd2(dba)3 (18.5 mg, 20.2 μmol, 0.04 eq) were dissolved in dioxane (4 mL), and then stirred at 140 °C for 1 h under microwave conditions. The MS value of the product was monitored by LCMS (RT = 0.347 min). The reaction solution was filtered, and the obtained filtrate was concentrated to obtain a crude product. The crude product was first purified by reverse-phase preparation (column: Waters Xbridge 150*25 mm*5um; mobile phase: [water(NH4HCO3)-ACN]; gradient: 12%-42% B over 9 min) to obtain 6′-(((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl)amino)-2H-[1,3′-bipyridin]-2-one (4 batches were run in parallel by microwave, 83.1 mg, 211 μmol, 10.5% yield, 98.4% purity).

[0157] LCMS MS(ESI) m / z = 388.2 [M+1] + 。

[0158] 1 H NMR, (400 MHz, DMSO-d6): 88.61 - 8.55 (m, 1H), 7.92 (d, J = 2.8 Hz, 1H), 7.60 (dd, J = 1.6, 6.8 Hz, 1H), 7.47 (ddd, J = 2.1, 6.7, 9.1 Hz, 1H), 7.44 - 7.34 (m, 2H), 6.92 (d, J = 6.9 Hz, 1H), 6.85 (dt, J = 1.6, 6.8 Hz, 1H), 6.64 (d, J = 7.3 Hz, 1H), 6.53 (d, J = 8.8 Hz, 1H), 6.44 (d, J = 8.8 Hz, 1H), 6.26 (dt, J = 1.3, 6.7 Hz, 1H), 4.40 - 4.26 (m, 1H), 4.18 (sxt, J = 6.7 Hz, 1H), 2.21 - 2.09 (m, 2H), 2.00 - 1.84 (m, 2H), 1.63 - 1.43 (m, 2H).

[0159] Among them, the deuterated compound C7A of compound 7 was prepared by combining this preparation method with the deuteration preparation method, and the structure is as follows:

[0160] The identification data is as follows: 1 H NMR (400MHz, DMSO-d6) δ8.59 (s, 1H), 7.92 (d, J=2.7Hz, 1H), 7.61 (dd, J=6.8, 2.1Hz, 1H), 7.53-7.4 6 (m, 1H), 7.46-7.33 (m, 3H), 6.94 (d, J = 6.9Hz, 1H), 6.66 (d, J = 7.3Hz, 1H), 6.53 (d, J = 8.9Hz, 1H), 6 .49-6.40 (m, 1H), 6.27 (td, J=6.7, 1.4Hz, 1H), 4.33 (q, J=6.6Hz, 1H), 4.19 (q, J=6.7Hz, 1H), 2.15 ( ddt, J=12.1, 7.0, 4.5Hz, 2H), 1.97 (dt, J=13.4, 6.8Hz, 1H), 1.93-1.83 (m, 1H), 1.65-1.40 (m, 2H).

[0161] Example C8

[0162] Synthesis of 6′-(((1S,3S)-3-((7-methoxy-[1,2,4]triazolo[1,5-a]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3′-bipyridyl]-2-one

[0163]

[0164] Step 1: Synthesis of 2,7-dibromo-[1,2,4]triazolo[1,5-a]pyridine

[0165] 7-Bromo-[1,2,4]triazolo[1,5-a]pyridin-2-amine (2.00 g, 9.39 mmol, 1.0 eq), copper bromide (3.15 g, 14.1 mmol, 1.5 eq), and tert-butyl nitrite (1.45 g, 14.1 mmol, 1.5 eq) were dissolved in acetonitrile (120 mL) and stirred at 80°C for 3 hours. LCMS (EW45820-76-P1A) detected the product (RT = 0.452 min). The reaction mixture was cooled to room temperature, diluted with water (200 mL), and extracted with ethyl acetate (80 mL x 2). The combined organic phases were washed with saturated brine (200 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to afford 2,7-dibromo-[1,2,4]triazolo[1,5-a]pyridine (2.00 g, 7.22 mmol, 76.9% yield). The product was used directly in the next step without purification. LCMS MS (ESI) m / z = 277.9 [M+1] + .

[0166] Step 2: Synthesis of 2-bromo-[1,2,4]triazolo[1,5-a]pyridin-7-ol

[0167] Dissolve 2,7-dibromo-[1,2,4]triazolo[1,5-a]pyridine (2.00 g, 7.22 mmol, 1.0 eq), potassium hydroxide (1.20 g, 21.7 mmol, 3.0 eq), t-Bu Xphos (675 mg, 1.59 mmol, 0.22 eq) and Pd2(dba)3 (728 mg, 794 μmol, 0.11 eq) in 1,4-dioxane (20 mL) and water (5 mL), and stir at 100 °C for 4 h. The MS value of the product was monitored by LCMS (RT = 0.358 min). Cool the reaction solution to room temperature, then dilute the reaction solution with water (60 mL) and ethyl acetate (40 mL), separate and discard the organic phase, adjust the pH of the aqueous phase to pH = 5 with dilute hydrochloric acid, and then extract the aqueous phase with ethyl acetate (60 mL × 2). The concentrated organic phase was washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 2-bromo-[1,2,4]triazolo[1,5-a]pyridin-7-ol (860 mg, 3.97 mmol, 54.9% yield, 98.7% purity). Without purification, it was directly used in the next step. LCMS MS (ESI) m / z = 215.9 [M+1]+.

[0168] Step 3: Synthesis of 2-bromo-7-methoxy-[1,2,4]triazolo[1,5-a]pyridine

[0169] Dissolve 2-bromo-[1,2,4]triazolo[1,5-a]pyridin-7-ol (1.30 g, 6.07 mmol, 1.0 eq) and cesium carbonate (4.95 g, 15.2 mmol, 2.5 eq) in N,N-dimethylformamide (20 mL), then add methyl iodide (1.72 g, 12.2 mmol, 2.0 eq) to the reaction solution, and stir at 40 °C for 2 h after addition. TLC (petroleum ether / ethyl acetate = 3 / 1) showed that the reaction was complete. Cool the reaction solution to room temperature, dilute the reaction solution with water (100 mL), and extract with ethyl acetate (30 mL × 2). The concentrated organic phase was washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was purified by silica gel column to obtain 2-bromo-7-methoxy-[1,2,4]triazolo[1,5-a]pyridine (1.00 g, 4.39 mmol, 72.2% yield).

[0170] Step 4: Synthesis of 6'-(((1S,3S)-3-((7-methoxy-[1,2,4]triazolo[1,5-a]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one

[0171] Dissolve 2-bromo-7-methoxy-[1,2,4]triazolo[1,5-a]pyridine (100 mg, 439 μmol, 1 eq), 6′-(((1S,3S)-3-aminocyclopentyl)amino)-2H-[1,3′-bipyridin]-2-one (130 mg, 482 μmol, 1.1 eq), Xantphos (20.3 mg, 35.1 μmol, 0.08 eq), sodium phenoxide (76.4 mg, 658 μmol, 1.5 eq) and Pd2(dba)3 (16.1 mg, 17.5 μmol, 0.04 eq) in dioxane (5 mL), and then stir at 140 °C for 1 h under microwave conditions. The MS value of the product was monitored by LCMS (RT = 0.366 min). Filter the reaction solution, and concentrate the obtained filtrate to get the crude product. The crude product was first purified by reverse-phase preparation (acidic conditions) (column: Phenomenex luna C18 150*40 mm*15 um; mobile phase: [water(FA)-ACN]; gradient: 20%-50% B over 15 min), and then purified by reverse-phase preparation (basic conditions) (column: Waters Xbridge 150*25 mm*5 um; mobile phase: [water(ammonia hydroxide v / v)-ACN]; gradient: 0%-30% B over 10 min) to obtain 6′-(((1S,3S)-3-((7-methoxy-[1,2,4]triazolo[1,5-a]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3′-bipyridin]-2-one (8 batches were run in parallel by microwave, 69.9 mg, 166 μmol, 4.72% yield, 98.9% purity).

[0172] LCMS MS(ESI) m / z = 418.2 [M+1] + 。 11H NMR, (400 MHz, DMSO-d6): δ 8.39 (d, J = 7.4 Hz, 1H), 7.91 (d, J = 2.6 Hz, 1H), 7.59 (dd, J = 1.7, 6.8 Hz, 1H), 7.47 (ddd, J = 2.0, 6.7, 9.1 Hz, 1H), 7.39 (dd, J = 2.6, 8.9 Hz, 1H), 6.89 (d, J = 6.9 Hz, 1H), 6.81 (d, J = 2.6 Hz, 1H), 6.58 - 6.38 (m, 4H), 6.26 (dt, J = 1.3, 6.7 Hz, 1H), 4.31 (sxt, J = 6.4 Hz, 1H), 4.13 (sxt, J = 6.7 Hz, 1H), 3.82 (s, 3H), 2.19 - 2.08 (m, 2H), 1.99 - 1.82 (m, 2H), 1.62 - 1.44 (m, 2H).

[0173] Example C9

[0174] Synthesis of 1-(5-((((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl)amino)pyrazin-2-yl)pyridin-2(1H)-one

[0175]

[0176] Step 1: Synthesis of 2-bromo-[1,2,4]triazolo[1,5-a]pyridine

[0177] Dissolve [1,2,4]triazolo[1,5-a]pyridin-2-amine (1.90 g, 14.2 mmol, 1.0 eq), copper(II) bromide (4.75 g, 21.3 mmol, 1.5 eq) and tert-butyl nitrite (2.19 g, 21.3 mmol, 1.50 eq) in acetonitrile (30 mL), stir at 70 °C for 4 h. TLC (petroleum ether / ethyl acetate = 1 / 1) shows the reaction is complete. Cool the reaction mixture to room temperature, then dilute it with water (50 mL) and extract with ethyl acetate (30 mL × 2). Wash the combined organic phases with saturated brine (100 mL × 2), dry over anhydrous sodium sulfate, filter, and concentrate to obtain 2-bromo-[1,2,4]triazolo[1,5-a]pyridine (2.17 g, 11.0 mmol, 77.4% yield). Use it directly in the next step without purification. LCMS MS (ESI) m / z = 200.0 [M+1] + 。

[0178] Step 2: Synthesis of 1-(5-((((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl)amino)pyrazin-2-yl)pyridin-2(1H)-one

[0179] 2-Bromo-[1,2,4]triazolo[1,5-a]pyridine (300 mg, 1.51 mmol, 1.0 eq), 1-(5-((((1S,3S)-3-aminocyclopentyl)amino)pyrazin-2-yl)pyridin-2(1H)-one (411 mg, 1.51 mmol, 1.0 eq), sodium tert-butoxide (437 mg, 4.54 mmol, 3.0 eq) and tBuXPhos PdG3 (120 mg, 152 μmol, 0.10 eq) were dissolved in dioxane (10 mL), and then stirred at 100 °C for 12 h. The MS value of the product was monitored by LCMS (RT = 0.394 min). The reaction solution was filtered, and the obtained filtrate was concentrated to obtain a crude product, which was purified by reverse-phase preparation (column: Phenomenex luna C18 150*40 mm*15um; mobile phase: [water(FA)-ACN]; gradient: 10%-40% B over 15 min) to obtain 1-(5-((((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl)amino)pyrazin-2-yl)pyridin-2(1H)-one (79.6 mg, 201 μmol, 13.2% yield, 97.9% purity).

[0180] LCMS MS(ESI) m / z = 389.2 [M+1] + 。 1 H NMR, (400 MHz, DMSO-d6): δ 8.62 - 8.54 (m, 1H), 8.25 (d, J = 1.4 Hz, 1H), 7.87 (d, J = 1.3 Hz, 1H), 7.72 (dd, J = 1.5, 6.9 Hz, 1H), 7.56 - 7.32 (m, 4H), 6.87 (dt, J = 1.5, 6.8 Hz, 1H), 6.66 (d, J = 7.3 Hz, 1H), 6.47 (d, J = 9.0 Hz, 1H), 6.32 (dt, J = 1.3, 6.7 Hz, 1H), 4.41 - 4.15 (m, 2H), 2.25 - 2.11 (m, 2H), 2.06 - 1.86 (m, 2H), 1.67 - 1.47 (m, 2H).

[0181] Example C10

[0182] Synthesis of 1-[5-[[[(1S,3S)-3-[(4-oxo-6,7-dihydro-5H-[1,2,4]triazolo[5,1-b][1,3]thiazin-2-yl)amino]cyclopentyl]amino]pyrazin-2-yl]pyridin-2-one

[0183]

[0184] Step 1: 2-Bromo-6,7-dihydro-5H-[1,2,4]triazolo[5,1-b][1,3]thiazine

[0185] Dissolve compound 10-1 (3.00 g, 13.2 mmol, 1.00 eq), 3-chloropropane-1-thiol (1.76 g, 15.8 mmol, 1.55 mL, 1.20 eq), and K2CO3 (3.60 g, 26.5 mmol, 2.00 eq) in DMF (20 mL), and stir the reaction solution at 50 °C for 5 hours.

[0186] The target product was detected by LCMS. Quench the reaction solution by adding 200 ml of water, then extract with ethyl acetate (100 * 3 mL). Combine the organic phases, wash the concentrated organic phase with saturated brine (100 mL * 2), dry over anhydrous sodium sulfate, filter, and concentrate. Purify the solid organic phase reaction solution using a normal-phase silica gel column (petroleum ether:ethyl acetate = 1:1) to obtain compound 10-2 (800 mg, 3.63 mmol, yield: 27.5%) as a white solid.

[0187] Step 2: Synthesis of 1-[5-[[[(1S,3S)-3-(6,7-dihydro-5H-[1,2,4]triazolo[5,1-b][1,3]thiazin-2-ylamino)cyclopentyl]amino]pyrazin-2-yl]pyridin-2-one

[0188] Under nitrogen protection, dissolve compound 10-3 (300 mg, 1.11 mmol, 1.00 eq), compound 10-2 (4988 mg, 2.27 mmol, 2.05 eq), t-BuONa (318 mg, 3.32 mmol, 3.00 eq), [2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladium; di-tert-butyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphine (175 mg, 221 μmol, 0.20 eq) in 1,4-dioxane (20 mL), and stir the reaction solution at 90 °C for 16 hours.

[0189] The target product was detected by LCMS and HPLC. The reaction mixture was quenched by adding 200 mL of water, and then extracted with ethyl acetate (100 * 3 mL). The organic phases were combined, and the combined organic phase was washed with saturated brine (100 mL * 2), dried over anhydrous sodium sulfate, filtered and concentrated. The organic phase was purified by reverse preparation (column: Phenomenex Luna C18 150 * 25 mm * 10 um; mobile phase: [water (TFA)-ACN]; gradient: 12%-42% B over 11 min) to obtain the target compound (60.0 mg, 146 μmol, yield: 13.2%) as a pale yellow gummy compound. LC-MS: [M + H]+ = 411.

[0190] 1H NMR (400 MHz, Chloroform-d) δ 8.52 (s, 1H), 7.73 (d, J = 1.6 Hz, 1H), 7.62 (dd, J = 7.2, 2.1 Hz, 1H), 7.41 - 7.33 (m, 1H), 6.62 (d, J = 9.3 Hz, 1H), 6.33 - 6.21 (m, 1H), 5.01 (d, J = 6.8 Hz, 1H), 4.31 (q, J = 6.6 Hz, 1H), 4.22 - 4.04 (m, 4H), 3.23 - 3.06 (m, 2H), 2.43 - 2.22 (m, 4H), 2.07 - 1.99 (m, 2H), 1.63 - 1.49 (m, 3H).

[0191] Example C11

[0192] Synthesis of 1-[5-[[[(1S,3S)-3-[(4-oxo-6,7-dihydro-5H-[1,2,4]triazolo[5,1-b][1,3]thiazin-2-yl)amino]cyclopentyl]amino]pyrazin-2-yl]pyridin-2-one

[0193]

[0194] Compound 10 (20.0 mg, 48.7 μmol, 1.00 eq) was dissolved in THF (8.00 mL), and then NaIO4 (10.4 mg, 48.7 μmol, 2.70 μL, 1.00 eq) dissolved in H2O (3 mL) was added. The reaction mixture was stirred at 70 °C for 12 hours.

[0195] The target product was detected by LCMS and HPLC. The reaction solution was directly concentrated under reduced pressure. The compound was purified by reverse preparation (column: Phenomenex Luna C18 150*25mm*10um; mobile phase: [water(TFA)-ACN]; gradient: 10%-40% B over 11min) to obtain the target compound (18.0 mg, 42.2 μmol, yield: 86.6%) as a white solid. LC-MS: [M-H]- = 427.1.

[0196] 1 1H NMR, (CHLOROFORM-d, 400 MHz): 8.54 (dd, 1H, J = 1.0, 4.3 Hz), 8.04 (d, 1H, J = 5.6 Hz), 7.7 - 7.8 (m, 1H), 7.52 (ddd, 1H, J = 1.9, 6.9, 9.0 Hz), 6.81 (d, 1H, J = 9.1 Hz), 6.46 (dt, 1H, J = 0.9, 6.8 Hz), 4.3 - 4.4 (m, 2H), 4.1 - 4.2 (m, 2H), 3.42 (dd, 1H, J = 5.9, 12.6 Hz), 2.9 - 3.2 (m, 2H), 2.3 - 2.5 (m, 3H), 2.0 - 2.2 (m, 2H), 1.6 - 1.8 (m, 2H).

[0197] Example C12

[0198] Referring to the preparation methods of the foregoing Examples C1 and C3, compounds C12, C12A, and C12B were prepared. The synthetic route is as follows:

[0199]

[0200] Compound 12 was subjected to chiral resolution (chromatographic column model: CHIRALCEL OD-H 5um 10mm*250mm; mobile phase: n-hexane / ethanol = 70:30; flow rate: 5 ml / min; column temperature was 30 °C) to obtain white solids C12A and C12B:

[0201] Compound C12A, 11H NMR: (400 MHz, DMSO-d6): δ 8.59 - 8.33 (m, 1H), 8.08 (dd, J = 1.6, 4.8 Hz, 1H), 7.92 (d, J = 2.4 Hz, 1H), 7.66 (dd, J = 1.4, 8.0 Hz, 1H), 7.55 - 7.48 (m, 2H), 7.40 (dd, J = 2.8, 8.9 Hz, 1H), 7.24 (dd, J = 4.8, 8.0 Hz, 1H), 6.96 (d, J = 6.9 Hz, 1H), 6.53 (d, J = 8.9 Hz, 1H), 6.31 (t, J = 6.8 Hz, 1H), 5.06 (br s, 1H), 4.79 - 4.62 (m, 1H), 4.37 (dt, J = 6.4, 13.8 Hz, 2H), 2.27 - 2.12 (m, 2H), 2.04 - 1.90 (m, 2H), 1.65 - 1.48 (m, 2H), 1.26 (d, J = 6.4 Hz, 3H);

[0202] Compound C12B, 1 1H NMR: (400 MHz, DMSO-d6): δ 8.54 - 8.37 (m, 1H), 8.08 (dd, J = 1.4, 4.8 Hz, 1H), 7.92 (d, J = 2.6 Hz, 1H), 7.66 (dd, J = 1.4, 8.1 Hz, 1H), 7.55 - 7.48 (m, 2H), 7.40 (dd, J = 2.6, 8.9 Hz, 1H), 7.24 (dd, J = 4.8, 8.0 Hz, 1H), 6.96 (d, J = 6.9 Hz, 1H), 6.53 (d, J = 89 Hz, 1H), 6.31 (t, J = 6.8 Hz, 1H), 5.06 (d, J = 4.6 Hz, 1H), 4.75 - 4.67 (m, 1H), 4.44 - 4.29 (m, 2H), 2.27 - 2.13 (m, 2H), 2.03 - 1.91 (m, 2H), 1.66 - 1.49 (m, 2H), 1.25 (d, J = 6.4 Hz, 3H).

[0203] Example C13

[0204] Referring to the preparation methods of the foregoing Examples C1, C3 and C12, compounds C13, C13A and C13B were prepared:

[0205]

[0206] Compound C13A, 11H NMR: (400 MHz, DMSO-d6): δ 8.49 (brd, J = 6.4 Hz, 1H), 8.16 - 8.08 (m, 1H), 7.89 (d, J = 2.0 Hz, 1H), 7.70 (dd, J = 1.2, 8.0 Hz, 1H), 7.62 - 7.55 (m, 3H), 7.28 (dd, J = 4.8, 8.0 Hz, 1H), 7.03 (brd, J = 7.2 Hz, 1H), 6.37 (t, J = 6.8 Hz, 1H), 5.12 (d, J = 4.4 Hz, 1H), 4.76 (quin, J = 5.6 Hz, 1H), 4.66 - 4.53 (m, 1H), 4.51 - 4.39 (m, 1H), 2.31 - 2.16 (m, 2H), 2.11 - 2.02 (m, 2H), 1.74 - 1.60 (m, 2H), 1.30 (d, J = 6.4 Hz, 3H);

[0207] Compound C13B, 1 1H NMR: (400 MHz, DMSO-d6): δ 8.45 (brd, J = 4.4 Hz, 1H), 8.08 (dd, J = 1.2, 4.8 Hz, 1H), 7.84 (d, J = 2.0 Hz, 1H), 7.66 (dd, J = 1.2, 8.0 Hz, 1H), 7.60 - 7.51 (m, 3H), 7.24 (dd, J = 4.8, 8.0 Hz, 1H), 6.98 (br d, J = 6.8 Hz, 1H), 6.33 (t, J = 6.8 Hz, 1H), 5.08 (d, J = 44 Hz, 1H), 4.80 - 4.65 (m, 1H), 4.55 (sxt, J = 7.2 Hz, 1H), 4.46 - 4.35 (m, 1H), 2.27 - 2.15 (m, 2H), 2.02 (t, J = 7.2 Hz, 2H), 1.69 - 1.56 (m, 2H), 1.26 (d, J = 6.4 Hz, 3H).

[0208] Example C14

[0209] Referring to the preparation method of the foregoing Example C2, Compound C14 was prepared:

[0210]

[0211] Example C17

[0212] Synthesize 6'-((((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl)amino)-5'-fluoro-2H-[1,3'-bipyridin]-2-one

[0213]

[0214] Step 1: Synthesis of tert-butyl N-[(1S,3S)-3-[(5-bromo-3-fluoro-2-pyridinyl)amino]cyclopentyl]carbamate

[0215] Dissolve tert-butyl N-[(1S,3S)-3-aminocyclopentyl]carbamate (3.00 g, 15.0 mmol, 1 eq), 5-bromo-2,3-difluoropyridine (2.91 g, 15.0 mmol, 1.0 eq) and N,N-diisopropylethylamine (4.84 g, 37.5 mmol, 2.5 eq) in dimethyl sulfoxide (40 mL), and stir at 100 °C for 12 h. The MS value of the product (RT = 0.568 min) was monitored by LCMS (EW45820-218-P1A1). The reaction solution was cooled to room temperature, then diluted with water (200 mL) and extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with saturated brine (500 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain tert-butyl N-[(1S,3S)-3-[(5-bromo-3-fluoro-2-pyridinyl)amino]cyclopentyl]carbamate (4.00 g). LCMS: RT = 0.568 min, MS(ESI) m / z = 374.0 [M+1] +

[0216] Step 2: Synthesis of tert-butyl N-[(1S,3S)-3-[[3-fluoro-5-(2-oxo-1-pyridinyl)-2-pyridinyl]amino]cyclopentyl]carbamate

[0217] tert-Butyl N-[(1S,3S)-3-[(5-bromo-3-fluoro-2-pyridinyl)amino]cyclopentyl]carbamate (1.5 g, 4.01 mmol, 1.0 eq), 1H-pyridin-2-one (457 mg, 4.81 mmol, 1.2 eq), potassium phosphate (2.55 g, 12.0 mmol, 3.0 eq), copper(I) iodide (153 mg, 802 μmol, 0.20 eq) and (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (114 mg, 802 μmol, 0.2 eq) were dissolved in dioxane (20 mL) and stirred at 100 °C for 12 h. TLC (petroleum ether / ethyl acetate = 3 / 1) showed complete reaction. The reaction solution was cooled to room temperature, then diluted with water (100 mL) and extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give tert-Butyl N-[(1S,3S)-3-[[3-fluoro-5-(2-oxo-1-pyridinyl)-2-pyridinyl]amino]cyclopentyl]carbamate (1.00 g).

[0218] Step 3: Synthesis of 6’-((((1S,3S)-3-aminocyclopentyl)amino)-5’-fluoro-2H-[1,3’-bipyridin]-2-one

[0219] tert-Butyl N-[(1S,3S)-3-[[3-fluoro-5-(2-oxo-1-pyridinyl)-2-pyridinyl]amino]cyclopentyl]carbamate (1.00 g, 2.57 mmol, 1.0 eq) was dissolved in dichloromethane (10 mL), then hydrochloric acid ethyl acetate HCl / EtOAc (2 M, 20 mL, 15.5 eq) was added thereto and stirred at 20 °C for 2 h. The MS value of the product (RT = 0.320 min) was monitored by LCMS (EW45820-225-P1A). The reaction solution was directly concentrated and dried to give 6′-((((1S,3S)-3-aminocyclopentyl)amino)-5′-fluoro-2H-[1,3′-bipyridin]-2-one (600 mg, 2.08 mmol, 80.8% yield). Without purification, it was directly used in the next step. LCMS: RT = 0.320 min, MS(ESI) m / z = 577.3 [2M+1] +

[0220] Step 4: Synthesis of 6’-((((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl)amino)-5’-fluoro-2H-[1,3’-bipyridin]-2-one

[0221] 6'-((((1S,3S)-3-Aminocyclopentyl)amino)-5'-fluoro-2H-[1,3'-bipyridin]-2-one (120 mg, 416 μmol, 1.0 eq), 2-bromo-[1,2,4]triazolo[1,5-a]pyridine (90.7 mg, 458 μmol, 1.1 eq), sodium tert-butoxide (120 mg, 1.25 mmol, 3.0 eq) and tBuXPhos Pd G3 (33.1 mg, 41.6 μmol, 0.1 eq) were dissolved in dioxane (4 mL), and then stirred in a microwave reactor at 140 °C for 1 hour. The MS value of the product (RT = 0.399 min) was monitored by LCMS (EW45820-233-P1A2). The reaction solution was filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by reverse-phase preparation twice (column: Phenomenex luna C18 150*40 mm*15 um; mobile phase: [water (FA)-ACN]; gradient: 15%-45% B over 15 min) to obtain 6'-((((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl)amino)-5'-fluoro-2H-[1,3'-bipyridin]-2-one (36.6 mg). LCMS: RT = 0.399 min, MS(ESI) m / z = 406.1 [M+1] + 。

[0222] Example C45

[0223] Synthesis of 6'-(((1S,3S)-3-((7,8-dihydro-[1,4]dioxo[2,3-d][1,2,4]triazolo[1,5-a]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one

[0224]

[0225] Step 1: Synthesis of 2-((6-bromo-4-iodopyridin-3-yl)oxy)ethan-1-ol

[0226] To a solution of C45-1 (8.00 g, 26.5 mmol, 1.00 eq) and ethylene glycol (55.6 g, 896 mmol, 50 mL, 33.8 eq) in NMP (50 mL) was added t-BuOK (5.95 g, 53.0 mmol, 2.00 eq). The mixture was stirred at 60 °C for 3 hours.

[0227] The target product was detected by LCMS. The reaction solution was poured into water, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness by rotary evaporation. The crude product was purified by reverse-phase column HPLC (0.1% FA condition) to obtain compound C45-2 (3.50 g, 10.1 mmol, 38.4% yield) as a white solid.

[0228] Step 2: Synthesis of 7-bromo-2,3-dihydro-[1,4]dioxino[2,3-c]pyridine

[0229] To a solution of C45-2 (3.50 g, 10.1 mmol, 1.00 eq) in isopropanol (35 mL) were added CuI (116 mg, 610 μmol, 0.06 eq), t-BuOK (1.60 g, 14.2 mmol, 1.40 eq), 3,4,7,8-tetramethyl-1,10-phenanthroline (192 mg, 814 μmol, 0.08 eq). Under nitrogen protection, the mixture was stirred at 80 °C for 1 h.

[0230] The target product was detected by LCMS. The reaction solution was poured into water, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness by rotary evaporation to obtain compound C45-3 (2.20 g, 9.12 mmol, 89.6% yield) as a brown solid.

[0231] Step 3: Synthesis of 2,3-dihydro-[1,4]dioxino[2,3-c]pyridin-7-amine

[0232] To a solution of C45-3 (2.20 g, 10.1 mmol, 1.00 eq) in ethylene glycol (20 mL) were added Cu2O (14.5 mg, 101 μmol, 10.4 μL, 0.01 eq), NH3·H2O (50.9 g, 407 mmol, 56.0 mL, 40.0 eq), K2CO3 (281 mg, 2.04 mmol, 0.200 eq), N′,N′-dimethyl-ethane-1,2-diamine (89.7 mg, 1.02 mmol, 111 μL, 0.100 eq). The mixture was stirred at 80 °C for 4 h.

[0233] The target product was detected by LCMS. The reaction solution was poured into water, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness by rotary evaporation to obtain compound C45-4 (1.50 g, crude) as a yellow oil.

[0234] Step 4: Synthesis of Ethyl N-(2,3-Dihydro-[1,4]dioxo[2,3-c]pyridin-7-ylaminothio)carbamate

[0235] To a solution of C45-4 (1.50 g, 9.86 mmol, 1.00 eq) in dichloromethane (15 mL) was added ethyl N-(thiomethylene)carbamate (1.29 g, 9.86 mmol, 1.00 eq). The mixture was stirred at 25 °C for 1 h.

[0236] The formation of the target product was detected by LCMS. The reaction solution was poured into water, extracted with dichloromethane, the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated and dried by rotary evaporation to obtain compound C45-5 (2.20 g, 7.77 mmol, 78.7% yield) as a yellow solid.

[0237] Step 5: Synthesis of 7,8-Dihydro-[1,4]dioxo[2,3-d][1,2,4]triazolo[1,5-a]pyridin-2-amine

[0238] To a solution of C45-5 (2.20 g, 7.77 mmol, 1.00 eq) in methanol (15 mL) was added NH2OH·HCl (2.43 g, 34.9 mmol, 4.50 eq) and DIEA (3.01 g, 23.30 mmol, 4.06 mL, 3 eq). The mixture was stirred at 70 °C for 2 h.

[0239] The formation of the target product was detected by LCMS. The reaction mixture was cooled to 25 °C, filtered, and the filter cake was compound C45-6 (1.30 g, 6.76 mmol, 87.1% yield) as a white solid.

[0240] Step 6: Synthesis of 2-Bromo-7,8-dihydro-[1,4]dioxo[2,3-d][1,2,4]triazolo[1,5-a]pyridine

[0241] At 0 °C, to a solution of C45-6 (500 mg, 2.60 mmol, 1.00 eq) in acetonitrile (5 mL) was added NaNO2 (359 mg, 5.20 mmol, 2.00 eq) and HBr (1.21 g, 5.98 mmol, 812 μL, 2.30 eq). The mixture was stirred at 25 °C for 2 h.

[0242] The target product was detected by LCMS. The reaction solution was poured into water, extracted with ethyl acetate, the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated and dried by rotary evaporation to obtain compound C45-7 (300 mg, 1.17 mmol, 45.0% yield) as a yellow solid.

[0243] Step 7: Synthesis of 6′-(((1S,3S)-3-((7,8-dihydro-[1,4]dioxo[2,3-d][1,2,4]triazolo[1,5-a]pyridin-2-yl)amino)cyclopentyl)amino)-2H-[1,3′-bipyridin]-2-one

[0244] To a solution of C45-7 (300 mg, 1.17 mmol, 1.00 eq) and C45-8 (316 mg, 1.17 mmol, 1.00 eq) in dioxane (5 mL) was added t-BuONa (225 mg, 2.34 mmol, 2.00 eq), Pd2(dba)3 (107 mg, 117 μmol, 0.100 eq) and Xantphos (135 mg, 234 μmol, 0.200 eq), and the mixture was stirred at 100 °C for 12 h under nitrogen protection.

[0245] The target product was detected by LCMS. The reaction solution was poured into water, extracted with ethyl acetate, the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated and dried by rotary evaporation. The crude product was purified by reverse preparation (column: Waters Xbridge Prep OBD C18 150*40 mm*10 um; mobile phase: [water (NH4HCO3)-ACN]; gradient: 10%-40% B over 52 min) and (column: Phenomenex luna C18 150*25 mm*10 um; mobile phase: [water (FA)-ACN]; gradient: 1%-25% B over 10 min) to obtain the target compound (36.6 mg).

[0246] LC-MS: [M+H] - = 446.1. 11H NMR: (DMSO-d6, 400 MHz): δ (ppm) = 8.38 (s, 1H), 7.91 (d, J = 2.4 Hz, 1H), 7.60 (dd, J = 2.0, 6.8 Hz, 1H), 7.49 - 7.45 (m, 1H), 7.39 (dd, J = 2.8, 8.8 Hz, 1H), 6.90 (d, J = 6.8 Hz, 1H), 6.78 (s, 1H), 6.52 (d, J = 9.2 Hz, 1H), 6.44 (d, J = 9.2 Hz, 1H), 6.36 (d, J = 7.2 Hz, 1H), 6.26 (dt, J = 1.2, 6.8 Hz, 1H), 4.37 - 4.24 (m, 5H), 4.18 - 4.05 (m, 1H), 2.20 - 2.06 (m, 2H), 1.99 - 1.78 (m, 2H), 1.64 - 1.41 (m, 2H).

[0247] Example C46

[0248] Synthesis of 6′ - ((((1S,3S)-3 - ((7,8 - dihydro - [1,4] dioxo[2,3 - d][1,2,4] triazolo[1,5 - a] pyridin - 2 - yl) amino) cyclopentyl) amino) - 5′ - fluoro - 2H - [1,3′ - bipyridin] - 2 - one

[0249]

[0250] Step 1: Synthesis of 2 - iodo - 7,8 - dihydro - [1,4] dioxo[2,3 - d][1,2,4] triazolo[1,5 - a] pyridine

[0251] At 0 °C, NaNO2 (215 mg, 3.12 mmol, 2.00 eq) and HI (1.02 g, 3.59 mmol, 600 μL, 2.30 eq) were added to a solution of C46 - 1 (300 mg, 1.56 mmol, 1.00 eq) in acetonitrile (5 mL). The mixture was stirred at 25 °C for 2 hours.

[0252] The formation of the target product was detected by LCMS. The reaction solution was poured into water, extracted with ethyl acetate, the organic phase was washed with an aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered, concentrated and evaporated to dryness to obtain compound C46 - 2 (480 mg, 1.11 mmol, 71.0% yield) as a brown solid.

[0253] Step 2: Synthesis of 6′-((((1S,3S)-3-((7,8-dihydro-[1,4]dioxo[2,3-d][1,2,4]triazolo[1,5-a]pyridin-2-yl)amino)cyclopentyl)amino)-5′-fluoro-2H-[1,3′-bipyridin]-2-one

[0254] To a solution of C46-2 (300 mg, 989 μmol, 1.00 eq) and C46-3 (285 mg, 989 μmol, 1.00 eq) in dioxane (5 mL) were added t-BuONa (190.26 mg, 1.98 mmol, 2 eq) and tBuBrettphos Pd G3 (84.5 mg, 98.9 μmol, 0.100 eq). Under nitrogen protection, the mixture was stirred at 100 °C for 4 hours.

[0255] The formation of the target product was detected by LCMS. The reaction solution was filtered, and the filtrate was concentrated and dried by rotary evaporation. The crude product was purified by reverse preparation (column: Waters Xbridge Prep OBD C18 150*40mm*10um; mobile phase: [water(NH4HCO3)-ACN]; gradient: 16%-46% B over 15 min) to obtain the target compound (20.0 mg).

[0256] LC-MS: [M+H] - = 464.1. 1 1H NMR: (DMSO-d6, 400 MHz): δ (ppm) = 8.38 (s, 1H), 7.84 (d, J = 2.0 Hz, 1H), 7.64 (dd, J = 1.6, 6.8 Hz, 1H), 7.57 - 7.45 (m, 2H), 6.90 (br d, J = 7.2 Hz, 1H), 6.78 (s, 1H), 6.46 (d, J = 8.8 Hz, 1H), 6.35 (d, J = 7.2 Hz, 1H), 6.28 (t, J = 6.4 Hz, 1H), 4.57 - 4.47 (m, 1H), 4.35 (br dd, J = 2.0, 5.2 Hz, 2H), 4.31 - 4.24 (m, 2H), 4.17 - 4.07 (m, 1H), 2.18 - 2.06 (m, 2H), 1.99 - 1.90 (m, 2H), 1.63 - 1.50 (m, 2H).

[0257] Example C47

[0258] Synthesis of 2-(6-(((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-

[0259]

[0260] Step A: tert-Butyl ((1S,3S)-3-((5-bromo-3-fluoropyridin-2-yl)amino)cyclopentyl)carbamate

[0261] At room temperature, 5-bromo-2,3-difluoropyridine (100.0 g, 0.52 mol) and tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (114.58 g, 0.57 mol) were added to DMF (800 mL). Subsequently, triethylamine (156.99 g, 1.55 mol) was added to the system, and then the temperature was raised to 100 °C and reacted for 8 hours.

[0262] After the reaction was completed, it was cooled to room temperature. Water (500 mL) was added to the system, and it was extracted with ethyl acetate (400 mL × 3). The organic phases were combined, washed twice with saturated brine (300 mL), dried, filtered, and concentrated under reduced pressure. The obtained residue was triturated with isopropyl ether (300 mL) to obtain 130 g of white solid product tert-butyl ((1S,3S)-3-((5-bromo-3-fluoropyridin-2-yl)amino)cyclopentyl)carbamate (yield 67%). LC-MS: [M+H]+ = 374.

[0263] Step B: tert-Butyl ((1S,3S)-3-((3-fluoro-5-(6-oxopyridazin-1(6H)-yl)pyridin-2-yl)amino)cyclopentyl)carbamate

[0264] At room temperature, tert-butyl ((1S,3S)-3-((5-bromo-3-fluoropyridin-2-yl)amino)cyclopentyl)carbamate (120 g, 0.32 mol), pyridazin-3(2H)-one (40 g, 0.41 mol), (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (9.1 g, 0.064 mol), and potassium carbonate (133.19 g, 0.96 mol) were added to NMP (960 mL). Subsequently, copper(I) iodide (12.3 g, 0.064 mol) was added. After addition, the nitrogen in the system was displaced and the temperature was raised to 120 °C and reacted for 12 hours.

[0265] After the reaction was completed, it was cooled to room temperature. Water (1000 mL) was added to the system, and the mixture was extracted with ethyl acetate (400 mL×3). The organic phases were combined, washed three times with saturated brine (400 mL), dried, filtered, and concentrated under reduced pressure. The resulting residue was purified by a normal-phase column (methylene chloride / methanol = 50:1) to obtain 70 g of a black oily product, tert-butyl ((1S,3S)-3-((3-fluoro-5-(6-oxopyridazin-1(6H)-yl)pyridin-2-yl)amino)cyclopentyl)carbamate (yield 56%). LC-MS: [M+H] + = 390.

[0266] Step C: 2-(6-(((1S,3S)-3-aminocyclopentyl)amino)-5-fluoropyridin-3-yl)pyridazin-3(2H)-one

[0267] At room temperature, tert-butyl ((1S,3S)-3-((3-fluoro-5-(6-oxopyridazin-1(6H)-yl)pyridin-2-yl)amino)cyclopentyl)carbamate (70 g, 0.17 mol) was dissolved in hydrochloric acid / 1,4-dioxane (2 M, 300 mL), and the reaction was carried out at room temperature for 4 hours.

[0268] After the reaction was completed, it was concentrated to dryness to obtain 2-(6-(((1S,3S)-3-aminocyclopentyl)amino)-5-fluoropyridin-3-yl)pyridazin-3(2H)-one hydrochloride, which was then dissolved in methanol (200 mL). Subsequently, the pH value of the system was adjusted to about 9 with a basic ion-exchange resin, filtered, and the filter cake was rinsed three times with methanol (100 mL). The filtrate was collected and then evaporated to dryness to obtain 55 g of a brown solid product, 2-(6-(((1S,3S)-3-aminocyclopentyl)amino)-5-fluoropyridin-3-yl)pyridazin-3(2H)-one, which was used directly in the next step without further treatment. LC-MS: [M+H] + = 290.

[0269] Step D: 2-(6-(((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl)amino)-5-fluoropyridin-3-yl)pyridazin-3(2H)-one

[0270] At room temperature, 2-(6-(((1S,3S)-3-aminocyclopentyl)amino)-5-fluoropyridin-3-yl)pyrazin-3(2H)-one (55 g, 0.19 mol) and 2-bromo-[1,2,4]triazolo[1,5-a]pyridine (37.4 g, 0.19 mol) were dissolved in 1,4-dioxane (550 mL), and then sodium tert-butoxide (36.5 g, 0.38 mol) and (2-(di-tert-butylphosphino)-2′,4′,6′-triisopropyl-1,1′-biphenyl)(2′-amino-1,1′-biphenyl-2-yl)palladium(II) (30 g, 0.037 mol) were added. After addition, nitrogen was displaced, and the system was slowly heated to 100 °C and reacted for 1 hour.

[0271] After the reaction was completed, it was cooled to room temperature. Water (400 mL) was added to the system, and it was extracted with ethyl acetate (300 mL × 3). The organic phases were combined, washed with saturated brine (200 mL), dried, filtered, and concentrated under reduced pressure. The residue was purified by a normal-phase column (dichloromethane / methanol = 10:1) to obtain 20.5 g of 2-(6-(((1S,3S)-3-([1,2,4]triazolo[1,5-a]pyridin-2-ylamino)cyclopentyl)amino)-5-fluoropyridin-3-yl)pyridazine-3(2H)-one. LC-MS: [M+H] + = 407.

[0272] 1 H NMR (400 MHz, DMSO-d6) δ 8.61 - 8.58 (dd, J = 6.7, 1.1 Hz, 1H), 8.06 - 8.04 (q, J = 1.7 Hz, 2H), 7.67 - 7.61 (dd, J = 12.1, 2.2 Hz, 1H), 7.52 - 7.47 (dd, J = 9.5, 3.9 Hz, 1H), 7.46 - 7.41 (m, 1H), 7.40 - 7.37 (dt, J = 8.8, 1.3 Hz, 1H), 7.09 - 7.05 (dd, J = 9.5, 1.6 Hz, 1H), 6.99 - 6.94 (m, 1H), 6.90 - 6.84 (td, J = 6.7, 1.6 Hz, 1H), 6.68 - 6.63 (d, J = 7.2 Hz, 1H), 4.61 - 4.51 (p, J = 7.0 Hz, 1H), 4.26 - 4.17 (p, J = 6.6 Hz, 1H), 2.22 - 2.12 (m, 2H), 2.02 - 1.97 (m, 2H), 1.67 - 1.54 (m, 2H).

[0273] Example C54

[0274] Synthesis of 2-(6-((((1S,3S)-3-((7,8-dihydro-[1,4]dioxo[2,3-d][1,2,4]triazolo[1,5-a]pyridin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)pyridazin-3(2H)-one

[0275]

[0276] Step 1: Synthesis of 2-(6-((((1S,3S)-3-((7,8-dihydro-[1,4]dioxo[2,3-d][1,2,4]triazolo[1,5-a]pyridin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)pyridazin-3(2H)-one

[0277] To a solution of C54-1 (120 mg, 395 μmol, 1.00 eq) and C54-2 (107 mg, 395 μmol, 1.00 eq) in dioxane (4 mL) were added t-BuONa (190 mg, 1.98 mmol, 5.00 eq) and tBuBrettphos PD G3 (33.8 mg, 39.60 μmol, 0.100 eq). Under nitrogen protection, the mixture was stirred at 100 °C for 12 h.

[0278] The formation of the target product was detected by LCMS. The reaction solution was filtered, and the filtrate was concentrated and dried by rotary evaporation. The crude product was purified by reverse-phase preparation (column: Phenomenex luna Cl8 150*25mm*10um; mobile phase: [water(FA)-ACN]; gradient: 1%-30% B over 10 min) to obtain the target compound (18.6 mg).

[0279] LC-MS: [M+H]- = 447.1. 1 H NMR: (DMSO-d6, 400 MHz): δ (ppm) = 8.37 (s, 1H), 8.09 (d, J = 2.4 Hz, 1H), 8.01 (dd, J = 1.6, 4.0 Hz, 1H), 7.51 (dd, J = 2.4, 8.8 Hz, 1H), 7.46 (dd, J = 4.0, 9.6 Hz, 1H), 7.02 (dd, J = 1.6, 9.6 Hz, 1H), 6.91 (d, J = 7.2 Hz, 1H), 6.77 (s, 1H), 6.52 (d, J = 8.8 Hz, 1H), 6.35 (d, J = 7.6 Hz, 1H), 4.37 - 4.25 (m, 5H), 4.16 - 4.07 (m, 1H), 2.18 - 2.08 (m, 2H), 1.99 - 1.83 (m, 2H), 1.56 - 1.44 (m, 2H).

[0280] Example C15 - C95

[0281] Referring to the aforementioned preparation method, compound C15 - C95 was prepared:

[0282]

[0283]

[0284]

[0285]

[0286]

[0287]

[0288]

[0289]

[0290] Related activity test of Example C96

[0291] Test method:

[0292] At 25 °C on Biacore TMSurface plasmon resonance data was collected on an 8K system (GE Healthcare). At 25 °C, NBS-N (10 mM HEPES, 0.15 M NaCl, pH 7.4) was used as the running buffer, and streptavidin was immobilized on a SA (Cytiva) sensor chip using standard amine coupling chemistry. Briefly, the carboxymethyl dextran surface was activated by injecting a 1:1 ratio of 0.4 M 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) / 0.1 M N-hydroxysuccinimide (NHS) at a flow rate of 10 μl / min for 12 min. To capture streptavidin, the protein was diluted to 0.5 mg / ml in 10 mM sodium acetate (pH 4.5) and captured by injecting 100 μl onto the activated chip surface. Excess residual activated groups were blocked by injecting 1 M ethanolamine (pH 8.5) for 7 min. 5 μl of protein diluted to 10 μg / mL in NBS-N, 0.05% Tween-20, 0.1 mM CaCl2 was injected at a flow rate of 5 μl / min for 60 s. The Aci-tagged PCSK9 protein was captured on the streptavidin surface. The typical surface density obtained was 2900 - 3200 RU. SPR binding data was obtained using an appropriate dilution series of each compound at a flow rate of 30 μl / min, with a capture time of 60 s / concentration point and a dissociation time of 3600 s. The running buffer used for compound binding studies was 10 mM HEPES, pH 7.4, 150 mM NaCl, 0.05% P20, 2% DMSO. Data excluding volume effects was corrected for DMSO. Using standard procedures, all data was double-referenced for blank injections and reference surfaces, and data processing and kinetic fitting were performed using Scrubber software version 2.0c (BioLogic software). A simple 1:1 binding model was used to fit the data to determine the K D value.

[0293] Test results:

[0294] K D is shown in Table 1 below:

[0295] Table 1. PCSK9 Kinetics data

[0296] Example <![CDATA[Kinetics K D (M)]]> C2 A C3 A

[0297] A < 1*E-8.

[0298] Example C97: Pharmacokinetic experiment

[0299] 1. Reagents and instruments

[0300] Polyethylene glycol 400 (batch number R22040588, Shanghai Shaoyuan Reagent Co., Ltd.), DMSO (batch number 20200319, Guangdong Guanghua Sci-Tech Co., Ltd.), normal saline (batch number 2011110727, Chenxin Pharmaceutical Co., Ltd.). LC-MS instruments (Thermo Fisher Ultimate 3000 UPLC, TSQ QUANTUM ULTRA triple quadrupole mass spectrometer, ABSCIEX 5500+QTARP).

[0301] 2. Experimental animals

[0302] SD rats: male, 180 - 250 g, purchased from Guangdong Vital River Laboratory Animal Technology Co., Ltd.

[0303] 3. Preparation of preparations

[0304] Precisely weigh the test sample powder. After completely dissolving it with DMSO, add PEG-400. After vortexing and ultrasonic mixing, add normal saline and vortex and ultrasonic mix to make it 0.5 mg / mL (DMSO:PEG-400:NS = 5:60:35, V / V / V). Administer by gavage at 10 mL / kg and by intravenous injection at 2 mL / kg.

[0305] 4. Blood sample collection

[0306] After intravenous or gavage administration to rats, collect 200 μL of venous blood at 5 min (not collected for gavage), 15 min, 30 min, 1 h, 2 h, 5 h, 7 h, and 24 h into EDTA-K2 anticoagulant EP tubes, centrifuge at 10000 rpm for 2 min, and take the plasma and store it at -80 °C for later measurement.

[0307] 5. Bioanalysis

[0308] Precisely weigh a certain amount of the test sample and dissolve it with DMSO to 2 mg / mL as the stock solution. Accurately pipette an appropriate amount of the compound stock solution and dilute it with acetonitrile to prepare a standard series of solutions. Accurately pipette 4 μL of each of the above standard series solutions, add 36 μL of blank plasma, vortex and mix well to prepare plasma samples with plasma concentrations equivalent to 1, 3, 5, 10, 30, 100, 300, 1000, and 3000 ng / mL. Analyze in duplicate for each concentration to establish a standard curve. Take 30 μL of plasma, add 200 μL of an acetonitrile solution of internal standard propranolol (5 ng / mL), vortex and mix well, then centrifuge at 4000 rpm for 10 min, and take the supernatant for LC-MS analysis. The LC-MS detection conditions are as follows:

[0309] Chromatographic column: YMC-Triart C18, 50×2.1 mm, S-3 μm 12 nm.

[0310] Mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile, flow rate: 0.5 mL / min, gradient elution is shown in Table 2 below:

[0311] Table 2

[0312] Time (min) A(％) B(％) 0 80％ 20％ 1.2 20％ 80％ 2.6 20％ 80％ 2.61 80％ 20％ 3.0 80％ 20％

[0313] 6. Data processing

[0314] After detecting the blood drug concentration by LC-MS, WinNonlin 6.1 software was used, and non-compartmental model was used to calculate the pharmacokinetic parameters of mice after drug administration. The results are shown in Table 3 below.

[0315] Table 3: Mouse pharmacokinetic parameters of the compounds of the present invention (iv and PO administration)

[0316] Compound Number Administration Method and Dose (mg / kg) t1 / 2 (h) AUC0-24h (h*ng / mL) C9 5, PO / 40300 C17 5, PO / 39800 C22 5, PO / 37700 C34 5, PO / 41400 C35 5, PO / 39700 C47 5, PO / 53000

[0317] The half-life and in vivo exposure of the compounds of the present invention are improved compared with the control compounds, and are superior to compounds 458B and 464 of CN113574055A, and compound C14.

[0318] Effect of Example C98 on hERG current in hERG-HEK293 cells[[ID=2S]]

[0319] Test method: Human embryonic kidney cells stably expressing hERG channels (hERG-HEK293 cells) were selected for the test. An automatic patch clamp system was used to clamp hERG-HEK293 cells to form a whole-cell voltage clamp mode, and hERG current was induced with corresponding voltages. The cells were respectively given extracellular fluid containing 0.3% DMSO (negative control) and 30 μM of the compound, or 1, 10, 100, and 1,000 nM of cisapride (positive control). The tail current of the hERG channel was recorded, and the peak value of the tail current at each concentration was obtained. Using the peak value of the tail current recorded under the negative control (0.3% DMSO) as 100%, the inhibition rates of 30 μM of the compound and different concentrations of cisapride on hERG current were calculated. The concentration-response relationship curve fitting of cisapride and IC 50 Calculation was completed using GraphPad Prism software, and the results are shown in Table 4.

[0320] Table 4 Effect on hERG current in hERG-HEK293 cells

[0321] Compound Number hERG Inhibition (%) Compound Number hERG Inhibition (%) Compound Number hERG Inhibition (%) C7 <50％ C30 <50％ C58 <50％ C8 <50％ C32 <50％ C61 <50％ C12 <50％ C34 <50％ C64 <50％ C13 <50％ C35 <50％ C65 <50％ C15 <50％ C36 <50％ C66 <50％ C17 <50％ C43 <50％ C67 <50％ C19 <50％ C47 <50％ C69 <50％ C21 <50％ C48 <50％ C71 <50％ C22 <50％ C53 <50％ C84 <50％ C26 <50％ C56 <50％

[0322] The Herg risk of the compounds of the present invention is improved compared with the control compounds, and is superior to the control compound 458B of CN113574055A.

[0323] Binding Affinity Test of Example C99 Compound and PCSK9 Protein

[0324] The compounds of the present invention were used to determine the binding affinity of the compounds for PCSK9 protein by fluorescence polarization method.

[0325] All compounds were dissolved in DMSO to prepare a stock solution of 10 mM. The positive compound and the compound to be tested started from 10 mM and were serially diluted 5-fold with DMSO, with a total of 8 concentration gradients. First, a certain volume of fluorescent probe solution was prepared using the test buffer (20 mM HEPES, 150 mM NaCl, 1 mM CaCl2, and 0.01% Tween-20) to make its concentration 5 nM; then the series of DMSO solutions of the test compounds were diluted 50-fold with the fluorescent probe solution; finally, a solution of human recombinant PCSK9 protein (ACRO, Cat#PC9-H5223) with a concentration of 4.5 μg / mL was prepared using the test buffer. After the test solution was prepared, 5 μL of PCSK9 protein was added to a black 384-well plate (PerkinElmer, Cat#6008260), and 5 μL of compounds with different concentrations (the final concentration of DMSO was 1%) was added. At the same time, a positive control group (test buffer + equal volume of target protein + equal proportion of fluorescent probe molecules) and a negative control group (test buffer + equal proportion of fluorescent probe molecules) were set. The final concentration of the probe molecule in the system was 2.5 nM, and the medium concentration of PCSK9 protein was 2.25 μg / mL. After shaking and incubating at room temperature for 15 minutes, the fluorescence polarization value was read using a microplate reader at an excitation wavelength of 485 nm and an emission wavelength of 520 nm. The inhibition rate of the drug = [1 - (mP (药物筛选组) - mP (阴性对照组) ÷ [mP (阳性对照组) - mP (阴性对照组) )] × 100. Using the logarithm of the compound concentration as the abscissa and the inhibition rate as the ordinate, a four-parameter nonlinear regression curve fitting was performed to calculate the IC 50 value (Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC A50 - X) * HillSlope)), where: Hillslope represents the slope of this curve, and IC 50 represents the half inhibitory concentration.

[0326] Table 5. FP-IC50 Data

[0327]

[0328]

[0329] B < 300 nm. * Incubated for 18 h.

[0330] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A compound represented by the general formula (C), or an isomer, or a racemate, or a pharmaceutically acceptable salt thereof, characterized in that: Ring A is selected from wherein X and Y form a 5- to 7-membered saturated or unsaturated ring, the 5- to 7-membered saturated or unsaturated ring contains 0, 1, and 2 heteroatoms, and the heteroatoms are selected from O, N, and S; B is selected from Q is selected from N or CR1, and R1 is selected from H or a halogen; T1 is selected from N or CH; wherein, R2 is selected from H, an alkyl group or a halogen, and there is one or more R2; R3 is selected from hydrogen or represents that the hydrogen on the A ring is further substituted by oxo, alkyl, halogen, alkoxy, alkylthio, haloalkyl, haloalkoxy, cycloalkyl, cycloalkylalkyl, alkynyl, R3 is one or more, or adjacent R3s form an alkoxy R4 is selected from hydrogen, halogen, hydroxy, alkoxy, haloalkoxy, substituted or unsubstituted alkyl, cyano, -C(O)-O-alkyl, phenylalkoxy, carboxyl, hydroxymethyl or cycloalkyl, the substituent is selected from hydroxy, amide, halogen, or substituted, U1, U2, U4 are independently selected from CH or N, U3 is selected from CH2 or NH, and R4 is one or more; and when ring A is selected from ring B is not 2. The compound according to claim 1, or an isomer, or a racemate, or a pharmaceutically acceptable salt thereof, characterized in that the alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, 1-ethylpropyl, 2-methylbutyl, tert-pentyl, 1,2-dimethylpropyl, isopentyl, neopentyl, n-hexyl, isohexyl, sec-hexyl, tert-hexyl, neohexyl, 2-methylpentyl, 1,2-dimethylbutyl, 1-ethylbutyl; the alkoxy group is selected from methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy; the cycloalkyl group is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; the halogen is selected from fluorine, chlorine, bromine, iodine.

3. The compound according to claim 1, or an isomer, or a racemate, or a pharmaceutically acceptable salt thereof, characterized in that, Q is selected from N; and / or Q is selected from CH and R2 is selected from F; and / or T1 is selected from N; and / or R3 is selected from hydrogen, methyl, methoxy, cyclopropyl, cyclopropylmethyl, fluorine, chlorine, oxo, CHF2-O-; R4 is selected from hydrogen, methyl, hydroxy, hydroxymethyl, cyano, F, Cl, Br, -O-CH2-benzene, -COOH, -COOCH2CH3, amide, formamide, CH3-C(O)-, ethynyl, trifluoromethyl, difluoromethoxy, methoxy.

4. The compound according to claim 1, or its isomer, or its racemate, or its pharmaceutically acceptable salt, characterized in that, Ring A is selected from Further, the A ring substituted by R3 is selected from: Furthermore, the B ring substituted by R4 is selected from:

5. The compound according to claim 1, or an isomer, or a racemate, or a pharmaceutically acceptable salt thereof, characterized in that, Selected from:

6. A pharmaceutical composition, characterized in that, Comprising a therapeutically effective amount of the compound according to any one of claims 1-5, or an isomer, or a racemate, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

7. The pharmaceutical use of the compound according to any one of claims 1-5, or an isomer, or a racemate, or a pharmaceutically acceptable salt thereof, specifically, the use in the preparation of a drug for treating a disease, wherein the disease is a PCSK9 inhibitor-related disease, preferably selected from conditions such as hypercholesterolemia.

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