An indazole derivative or a pharmaceutically acceptable salt thereof and use thereof

Abstract

The application discloses an indazole derivative or a pharmaceutical salt thereof and application. The application finds a new indazole derivative, which has an inhibiting function on ROCK kinase, can be used as a ROCK kinase inhibitor, and has a novel structure type, and can be used as a potential therapeutic drug for various diseases, such as inflammatory diseases, autoimmune diseases, fibrotic diseases, transplant rejection, diseases related to excessive secretion of IL-17, IL-21 and / or IL-23, eye diseases, cardiovascular and cerebrovascular diseases, tumor metastasis, central nervous system diseases, diabetic nephropathy and skin diseases, and the like.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, and in particular to an indazole derivative or a pharmaceutical salt thereof and application. BACKGROUND

[0002] Rho protein-related spiral kinase (ROCK) is a serine / threonine protein kinase of the AGC kinase family, which is a downstream effector of Rho. ROCK kinase protein contains an N-terminal kinase domain, a Rho protein binding domain (RBD), and a PH domain divided by a C-terminal zinc finger motif domain (CRD). In the classic ROCK signaling pathway, Rho GTPase activates downstream ROCK kinase, and further phosphorylates the downstream substrate of ROCK (MLC, LIMK, ERM, MARCKS, CRMP-2, etc.), remodeling the cytoskeleton, inducing actin-myosin contraction, and regulating microtubule dynamics. ROCK kinase is composed of two subtypes, ROCK1 and ROCK2, which have 65% homology in amino acid sequence and 92% similarity in amino acid sequence in the kinase domain. ROCK is distributed in tissues throughout the body. Compared with ROCK2, ROCK1 is more highly expressed in non-neural tissues such as blood, small intestine, thymus, etc., and ROCK2 is more highly expressed in brain, heart and colon, etc.

[0003] ROCK kinase is involved in the occurrence and development of various diseases, such as eye diseases, cardiovascular and cerebrovascular diseases, tumor metastasis, central nervous system diseases, diabetic nephropathy, etc., and is an important target for treating various diseases. Rho GTPase-mediated signaling pathways play an important role in balancing and coordinating T cell-mediated immune processes, including T cell development, activation and differentiation. Recent studies have shown that overactivation of ROCK kinase also plays an indispensable role in inflammatory diseases and autoimmune diseases, such as psoriasis, inflammatory bowel disease, rheumatoid arthritis, systemic lupus erythematosus, etc. Although the dysregulated ROCK kinase mainly affects the autoimmune process by regulating the dynamic cytoskeleton, ROCK2 can regulate autoimmunity by controlling the balance of pro-inflammatory and anti-inflammatory T cell populations. Recent reports show that ROCK2 regulates the production of IL-21 and IL-17 by phosphorylating interferon regulatory factor 4 (IRF4) in mice, and plays an important role in the development of autoimmunity. In human peripheral blood CD4 +In Th17 cells, ROCK2 binds to phosphorylated STAT3 to regulate the transcription of Th17 cell-specific genes. A clinical study showed that oral administration of a selective ROCK2 inhibitor (KD025) can reduce the secretion of IL-17 and IL-21 induced by external stimuli in healthy volunteers (Natl. Acad. Sci. 2014, 111, 16814-16819). Another two studies showed that targeting ROCK2 can down-regulate over-activated Th17 cells and enhance the function of regulatory T cells (Treg) by inhibiting STAT3 phosphorylation and increasing STAT5 phosphorylation, thereby re-establishing immune balance (Blood. 2016, 127, 2144-2154). Tenggesdal et al. showed that KD025 down-regulates IL-17 secretion independent of IL-1 and IL-6 (Eur. J. Immunol. 2018, 48, 1679-1686). In non-hematopoietic cells, ROCK proteins are also involved in the TGF-β signaling pathway that controls the balance between Th17 and Treg cells (Nat. Commun. 2020, 11, 2608). Based on the role of ROCK2 in inhibiting the over-activation of Th17 cells, it is expected to become a new target for the treatment of psoriasis.

[0004] So far, only four small molecule ROCK kinase inhibitors have been approved for marketing. Fasudil is the first approved ROCK inhibitor with a Ki value of 0.33 μM for inhibiting ROCK2, and is mainly used for the treatment of glaucoma, cerebral vasospasm and cerebral ischemia, etc., and also has good effects in the treatment of amyotrophic lateral sclerosis (ALS), induction of tumor cell apoptosis, inhibition of tumor metastasis, and reduction of symptoms of nervous system diseases (such as Parkinson's disease). Ripasudil was approved for marketing in Japan in 2014 and is mainly used for the treatment of glaucoma and high intraocular pressure, with IC50 values of 19 and 51 nM for inhibiting ROCK2 and ROCK1, respectively. Netarsudil was approved by the FDA in 2017 for the treatment of glaucoma or high intraocular pressure, with a Ki value of 2 nM for inhibiting ROCK2. KD025 is the most selective inhibitor of ROCK2 subtype kinase, with an IC50 value of 24 μM for inhibiting ROCK1 and an IC50 value of 105 nM for inhibiting ROCK2. In 2021, the compound was approved by the FDA for the treatment of anti-graft-versus-host disease. In addition, there are seven compounds in the clinical research stage, mainly for the treatment of glaucoma, spinal cord injury and pulmonary fibrosis, etc. SUMMARY

[0005] The purpose of the present application is to provide an indazole derivative or a pharmaceutically acceptable salt thereof, which has an inhibitory function on ROCK kinase and can be used as a ROCK kinase inhibitor.

[0006] The present application provides, at first, an indazole derivative or a pharmaceutically acceptable salt thereof, the structure of which is shown in the general formula (I),

[0007]

[0008] wherein:

[0009] G is CH or N atom;

[0010] Ring A is selected from 5- to 7-membered aryl, heteroaryl, cycloalkyl and heterocyclyl;

[0011] Each R 1 is the same or different, and each is independently selected from hydrogen atom, halogen, alkyl group having 1 to 6 carbon atoms, haloalkyl group having 1 to 6 carbon atoms, haloalkoxy group having 1 to 6 carbon atoms, cyano group, amino group, hydroxyl group, nitro group, carboxyl group, hydroxyalkyl group having 1 to 6 carbon atoms, cycloalkyl group, heterocyclyl group, aryl group and heteroaryl group;

[0012] Each R 2 is the same or different, and each is independently selected from hydrogen atom, halogen, cyano group, hydroxyl group, nitro group, carboxyl group, hydroxyalkyl group having 1 to 6 carbon atoms;

[0013] Each R 3 is the same or different, and each is independently selected from hydrogen atom, halogen, alkyl group having 1 to 6 carbon atoms, haloalkyl group having 1 to 6 carbon atoms, haloalkoxy group having 1 to 6 carbon atoms, cyano group, hydroxyl group, nitro group, carboxyl group, hydroxyalkyl group having 1 to 6 carbon atoms;

[0014] a is 0, 1, 2, 3 or 4;

[0015] q is 0, 1 or 2;

[0016] s is 0, 1 or 2.

[0017] In the present application, the phrase "selected from" means selecting one from a plurality of groups.

[0018] Preferably, R 1 When alkyl group, haloalkyl group, haloalkoxy group or hydroxyalkyl group is selected, the number of carbon atoms is 1 to 4, which is straight or branched. When alkyl group is selected, methyl group, phenyl group, tert-butyl group or ethyl group is most preferred.

[0019] Preferably, R 2 When hydroxyalkyl group is selected, it is hydroxymethyl group.

[0020] Preferably, R 3 When alkyl group, haloalkyl group, haloalkoxy group or hydroxyalkyl group is selected, the number of carbon atoms is 1, i.e. methyl group, halomethyl group, halomethoxy group or hydroxymethyl group.

[0021] Preferably, the indazole derivative has a structure represented by general formula (I-1), (I-2), (I-3) or (I-4),

[0022]

[0023] Preferably, ring A is selected from 5- to 7-membered cycloalkyl, heterocyclyl and phenyl.

[0024] More preferably, ring A is selected from cyclopentane, cyclohexane, cycloheptane, phenyl and pyridyl, and the indazole derivative has a structure represented by general formula (II-1), (II-2) or (II-3),

[0025]

[0026] wherein n is 0, 1 or 2.

[0027] Further preferably, the indazole derivative has a structure represented by general formula (III-1), (III-2), (III-3), (III-4), (III-5), (III-6) or (III-7),

[0028]

[0029] Further preferably, the indazole derivative is any one of the following compounds:

[0030]

[0031]

[0032] wherein (S) and (R) represent chiral cis and trans, respectively.

[0033] The present application also provides use of the indazole derivative or a pharmaceutically acceptable salt thereof in the preparation of a ROCK kinase inhibitor.

[0034] The present application also provides use of the indazole derivative or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating and / or preventing a disease or a condition selected from inflammatory diseases, autoimmune diseases, fibrotic diseases, transplant rejection, diseases associated with overproduction of IL-17, IL-21 and / or IL-23, ocular diseases, cardiovascular and cerebrovascular diseases, tumor metastasis, central nervous system diseases, diabetic nephropathy and dermatological diseases.

[0035] Preferably, the disease or condition is selected from rheumatoid arthritis, osteoarthritis, juvenile idiopathic arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, allergic airway disease, chronic obstructive pulmonary disease, asthma, bronchitis, inflammatory bowel disease, systemic lupus erythematosus, cutaneous lupus erythematosus, lupus nephritis, dermatomyositis, autoimmune liver disease, Sjogren's syndrome, multiple sclerosis, dry eye disease, type I diabetes and complications associated therewith, atopic eczema, thyroiditis, contact dermatitis, Sjogren's syndrome and amyotrophic lateral sclerosis.

[0036] The present application also provides a medicament, the active ingredient of which is the indazole derivative or a pharmaceutically acceptable salt thereof.

[0037] The present application also provides a preparation method of the indazole derivative or a pharmaceutically acceptable salt thereof, the reaction formula of which is as follows:

[0038]

[0039] wherein the compound or salt represented by general formula (IA) and general formula (IV-1) undergoes reductive amination to obtain the compound or pharmaceutically acceptable salt represented by general formula (I).

[0040] The preparation method further comprises the preparation method of the compound represented by general formula (IA):

[0041]

[0042] wherein the compound or salt represented by general formula (IA-1) and general formula (IA-2) undergoes coupling reaction to obtain the compound or salt represented by general formula (IA).

[0043] The preparation method further comprises the preparation method of the compound represented by general formula (IV-1):

[0044]

[0045] wherein the compound or salt represented by general formula (V-1) undergoes deprotection reaction to obtain the compound or salt represented by general formula (IV-1).

[0046] The preparation method further comprises the preparation method of the compound represented by general formula (IV-1):

[0047]

[0048] wherein the compound or salt represented by general formula (IV-2) and general formula (IV-4) undergoes reductive amination to obtain the compound or salt represented by general formula (V-1). Alternatively, the compound or salt represented by general formula (IV-3) and general formula (IV-4) undergoes coupling reaction to obtain the compound or salt represented by general formula (V-1).

[0049] The present application discloses a new class of indazole derivatives, which have inhibitory function on ROCK kinase, can be used as ROCK kinase inhibitors, and have novel structure type, and can be used as potential therapeutic drugs for various diseases, such as inflammatory diseases, autoimmune diseases, fibrotic diseases, transplant rejection, diseases related to over-secretion of IL-17, IL-21 and / or IL-23, eye diseases, cardiovascular and cerebrovascular diseases, tumor metastasis, central nervous system diseases, diabetic nephropathy and skin diseases. DETAILED DESCRIPTION

[0050] The present application is further described below in conjunction with examples, but the examples are not intended to limit the scope of the present application. The indazole derivatives of the present application are any one of the following compounds:

[0051]

[0052]

[0053] Example 1

[0054] The compounds in Example 1 are obtained by the following reaction scheme:

[0055]

[0056] Step 1, synthesis of 1b:

[0057] Sodium hydride (567 mg, 23.6 mmol) was added to anhydrous tetrahydrofuran (80 mL) under ice bath environment, and benzyl alcohol (1.5 mL, 14.16 mmol) was added under nitrogen atmosphere, and stirred for 1 hour. Compound 1a with different substituents dissolved in anhydrous tetrahydrofuran (10 mL) was added, and reacted at 50°C for 18 hours. After cooling to room temperature, saturated ammonium chloride was added for quenching, and the solvent was removed by concentration under reduced pressure. The residual liquid was extracted with dichloromethane, and the organic phase was combined after being repeated three times and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with petroleum ether / ethyl acetate (40:1, v / v, 500 mL) to obtain the title compound 1b with a yield of 60-82%.

[0058] Step 2, synthesis of 1c:

[0059] Compound 1b with different substituents (6.45 mmol) and 10% Pd / C (68 mg, 10 mol%) were added to a round-bottom flask, and methanol (60 mL) and ethyl acetate (30 mL) were added. After stirring at room temperature for 2 hours under hydrogen atmosphere, the mixture was filtered through diatomite, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography with dichloromethane / methanol (20:1, v / v, 500 mL) to obtain the title compound 1c with a yield of 90-95%.

[0060] Step 3, synthesis of tert-butyl ((1S)-3-((1H-indazol-5-yl)amino)cyclohexyl)carbamate 1f:

[0061] To a solution of (S)-(3-oxocyclohexyl)carbamic acid tert-butyl ester 1d (Bide Pharmatech, CAS No.: 1803033-61-5) (1.00 g, 4.69 mmol), 5-aminoindazole 1e (Bide Pharmatech, CAS No.: 19335-11-6) (750 mg, 5.63 mmol), sodium cyanoborohydride (500 mg, 7.96 mmol) in methanol (50 mL) was added acetic acid to adjust pH to 6.0-7.0, and the reaction was stirred at room temperature for 12 hours. Water (2 mL) was added, and the solvent was removed by reduced pressure concentration. The residue was purified by silica gel column chromatography with dichloromethane / methanol (40:1, v / v, 500 mL) containing 1% triethylamine by volume percentage to give the title compound 1f (1.42 g), yield: 92%.

[0062] The obtained product 1f was analyzed by mass spectrometry, m / z (ESI): 331.2 [M+1].

[0063] Step 4, synthesis of (3S)-N 1 -(1H-indazol-5-yl)cyclohexyl-1,3-diamine 1g:

[0064] To a solution of compound 1f (1.42 g, 4.30 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (10 mL), and the reaction was stirred at room temperature for 1 hour. The reaction solution was concentrated by reduced pressure, and the residue was purified by silica gel column chromatography with dichloromethane / methanol (5:1, v / v, 500 mL) containing 1% triethylamine by volume percentage to give the title compound 1g (815.7 mg), yield: 82%.

[0065] The obtained product 1g was analyzed by mass spectrometry, m / z (ESI): 231.2 [M+1].

[0066] Step 5, synthesis of 1i:

[0067] To a solution of compound 1c (2.36 mmol) with different substituents, 3-formylphenylboronic acid 1h (Bide Pharmatech, CAS No.: 87199-16-4) (708 mg, 4.72 mmol), copper acetate (944 mg, 4.72 mmol), pyridine (4.5 mL) in dichloromethane (120 mL) was added, and the reaction was stirred at room temperature for 12 hours. The reaction solution was concentrated by reduced pressure, and the residue was purified by silica gel column chromatography with dichloromethane (500 mL) containing 1% triethylamine by volume percentage to give the corresponding compound 1i, yield: 54-91%.

[0068] Step 6, synthesis of II-1 and II-2:

[0069] To a solution of compound 1i (0.43 mmol), compound 1g (100 mg, 0.43 mmol), sodium cyanoborohydride (48 mg, 0.76 mmol) in methanol (20 mL) was added acetic acid to adjust pH to 6.0-7.0, and the reaction was stirred at room temperature for 12 hours. Water (1 mL) was added, and the solvent was removed by concentration under reduced pressure. The residue was purified by high performance liquid chromatography (separation condition: preparative column Welch XB-C18, 21.2 x 250 mm, 5 μm; elution system: methanol / water (volume ratio) = 10%-100% containing 0.5% volume fraction of trifluoroacetic acid, flow rate: 10 ml / min, elution time: 40 min) to give the corresponding compounds II-1 and II-2 with a yield of 30-32% and 61-63%, respectively. XB-C18, 21.2 x 250 mm, 5 μm; elution system: methanol / water (volume ratio) = 10%-100% containing 0.5% volume fraction of trifluoroacetic acid, flow rate: 10 ml / min, elution time: 40 min) to give the corresponding compounds II-1 and II-2 with a yield of 30-32% and 61-63%, respectively.

[0070] The single configuration compounds 12-1 and 12-2 were prepared from 2-chloro-5-fluoro-4-methylpyridine 1a (Bide Pharm, CAS No: 881891-83-4).

[0071] The single configuration compound 12-1: 1 H NMR (600 MHz, methanol-d4) δ 7.92 (s, 1H), 7.65 (d, J = 5.0 Hz, 1H), 7.60 (t, J = 7.8 Hz, 1H), 7.57-7.54 (m, 2H), 7.49-7.45 (m, 2H), 7.12 (d, J = 8.7 Hz, 1H), 6.53 (d, J = 6.9 Hz, 1H), 4.32 (d, J = 13.3 Hz, 1H), 4.27 (d, J = 13.3 Hz, 1H), 3.54-3.49 (m, 1H), 3.35-3.32 (m, 1H), 2.55-2.53 (m, 1H), 2.30 (s, 3H), 2.27-2.25 (m, 1H), 2.14-2.12 (m, 1H), 2.04-2.01 (m, 1H), 1.56-1.49 (m, 1H), 1.47-1.41 (m, 2H), 1.39-1.32 (m, 1H). 13C NMR (300 MHz, methanol-d4) δ 162.4, 149.8 (d, J = 232.6 Hz), 146.8 (d, J = 20.2 Hz), 141.9, 138.5, 137.5, 134.0, 133.9, 131.6, 131.4, 129.6, 128.8, 124.7, 124.2 (d, J = 39.0 Hz), 121.6, 121.0 (d, J = 2.9 Hz), 112.5, 106.9, 57.0, 55.4, 49.6, 35.2, 31.6, 29.5, 23.1, 15.3. MS m / z (ESI): 446.2 [M + 1].

[0072] Single configuration compound 12-2: 1 H NMR (600 MHz, methanol-d4) δ 7.82 (s, 1H), 7.60 (d, J = 4.6 Hz, 1H), 7.54 ((t, J = 7.8 Hz, 1H), 7.51-7.49 (m, 2H), 7.46-7.44 (m, 1H), 7.35 (d, J = 9.6 Hz, 1H), 6.98 (d, J = 8.7 Hz), 6.51 (dd, J = 7.3, 0.9 Hz), 4.29 (d, J = 13.3 Hz, 1H), 4.24 (d, J = 13.3 Hz, 1H), 3.92-3.91 (m, 1H), 3.54-3.49 (m, 1H), 2.48-2.45 (m, 1H), 2.29 (s, 3H), 2.20-2.18 (m, 1H), 1.89-1.77 (m, 3H), 1.76-1.69 (m, 2H), 1.60-1.54 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 162.4, 149.8 (d, J = 234.1 Hz), 146.8 (d, J = 20.2 Hz), 142.2, 141.9, 137.1, 133.8, 133.3, 131.6, 131.6, 129.6, 128.7, 125.0, 124.2 (d, J = 39.0 Hz), 121.1, 121.0 (d, J = 4.3 Hz), 112.1, 101.5, 54.5, 49.6, 48.6, 32.9, 30.3, 29.7, 20.2, 15.3. MS m / z (ESI): 446.2 [M + 1].

[0073] Single configuration compounds 15-1 and 15-2 were prepared from 4-(tert-butyl)-2- chloropyridine la (Bide Pharm, CAS number: 81167-60-4).

[0074] Single configuration compound 15-1: 1 H NMR (600 MHz, methanol-d4) δ 7.91 (s, 1H), 7.61 (t, J = 7.8 Hz, 1H), 7.57-7.55 (m, 1H), 7.54-7.52 (m, 2H), 7.48 (d, J = 9.2 Hz, 1H), 7.45-7.43 (m, 1H), 7.12 (d, J = 8.7 Hz, 1H), 6.63 (dd, J = 7.3, 2.3 Hz, 1H), 6.58 (d, J = 1.8 Hz, 1H), 4.33 (d, J = 13.3 Hz, 1H), 4.27 (d, J = 13.3 Hz, 1H), 3.54-3.50 (m, 1H), 3.35-3.31 (m, 1H), 2.56-2.53 (m, 1H), 2.27-2.25 (m, 1H), 2.14-2.11 (m, 1H), 2.04-2.01 (m, 1H), 1.53-1.50 (m, 1H), 1.47-1.41 (m, 2H), 1.38-1.33 (m, 1H), 1.31 (s, 9H). 13 C NMR (300 MHz, methanol-d4) δ 167.7, 164.8, 142.3, 138.5, 138.5, 137.4, 134.0, 133.9, 131.6, 131.3, 129.7, 128.8, 124.8, 121.7, 116.1, 112.5, 108.3, 107.0, 57.0, 55.4, 49.6, 36.1, 35.3, 31.7, 29.9, 29.5, 23.1. MS m / z (ESI): 470.3 [M+1].

[0075] Single configuration compound 15-2: 1 H NMR (600 MHz, methanol-d4) δ 7.85 (s, 1H), 7.55 (t, J = 7.8 Hz, 1H), 7.52-7.49 (m, 2H), 7.47 (d, J = 8.2 Hz, 1H), 7.45-7.43 (m, 1H), 7.39 (d, J = 8.7 Hz, 1H), 7.03 (d, J = 9.2 Hz, 1H), 6.58-6.56 (m, 2H), 4.30 (d, J = 13.3 Hz, 1H), 4.26 (d, J = 13.3 Hz, 1H), 3.94-3.93 (m, 1H), 3.57-3.53 (m, 1H), 2.44-2.42 (m, 1H), 2.20-2.18 (m, 1H), 1.89-1.74 (m, 5H), 1.63-1.58 (m, 1H), 1.30 (s, 9H). 13C NMR (300 MHz, methanol-d4) δ 167.7, 164.8, 142.3, 141.1, 138.4, 137.4, 133.8, 133.5, 131.6, 131.4, 129.6, 128.8, 125.0, 121.2, 116.1, 112.2, 108.2, 101.9, 54.4, 50.6, 48.8, 36.1, 32.8, 30.0, 29.9, 29.5, 20.1. MS m / z (ESI): 470.3 [M+1].

[0076] Example 2

[0077] The compounds in Example 2 were obtained from the following reaction scheme:

[0078]

[0079] First step, synthesis of 1k:

[0080] The compound 1j (5.0 mmol) with different substituents, the compound 1h (10.0 mmol) with para or meta substituted aldehyde group, copper acetate monohydrate (998 mg, 5.0 mmol), tetramethyl ethylenediamine (1.5 mL) were added into dichloromethane (120 mL) and reacted at room temperature for 12 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with dichloromethane (500 mL) containing 1% triethylamine by volume percentage to obtain the title compound 1k (297.7 mg) with a yield of 63%.

[0081] Second step, synthesis of III-1, III-2, III-3, III-4, III-7 and III-8:

[0082] The corresponding compound 1k (100 mg, 0.43 mmol), compound 1g or 1g-1 (100 mg, 0.43 mmol), sodium cyanoborohydride (48 mg, 0.76 mmol) were added into anhydrous methanol (20 mL), and the pH was adjusted to 6.0-7.0 with acetic acid, and the reaction was carried out at room temperature for 12 hours. Water (1 mL) was added, and the solvent was removed by concentration under reduced pressure. The residue was purified by high performance liquid chromatography (separation conditions: preparative column Welch XB-C18, 21.2 x 250 mm, 5 μm; elution system: methanol / water (volume ratio) = 10%-100% containing 0.5% trifluoroacetic acid by volume, flow rate: 10 ml / min, elution time: 40 min) to obtain the corresponding compounds III-1, III-2, III-3, III-4, III-7 and III-8 with yields of 25-36% and 51-59%, respectively.

[0083] ​Single configuration compounds 1-1 and 1-2 were prepared from 5-fluorocytosine 1j (Bedoukian Research, CAS Number: 2022-85-7).

[0084] Single configuration compound 1-1: 1 H NMR (600 MHz, methanol-d4) δ 8.06 (s, 1H), 7.90 (d, J = 6.0 Hz, 1H), 7.65-7.61 (m, 2H), 7.56-7.52 (m, 2H), 7.48 (d, J = 7.8 Hz, 1H), 7.40 (dq, J = 7.8, 0.9 Hz, 1H), 7.30 (dd, J = 9.2, 2.3 Hz, 1H), 4.31 (d, J = 12.8 Hz, 1H), 4.24 d, J = 12.8 Hz, 1H), 3.65-3.59 (m, 1H), 3.32-3.28 (m, 1H), 2.45-2.40 (m, 1H), 2.39-2.24 (m, 1H), 2.15-2.11 (m, 1H), 2.08-2.03 (m, 1H), 1.60-1.40 (m, 4H). 13 C NMR (300 MHz, methanol-d4) δ 160.1 (d, J = 15.9 Hz), 156.4, 142.0, 139.8, 138.8 (d, J = 244.2 Hz), 134.7, 133.6, 131.6, 131.3 (d, J = 33.2 Hz), 130.9, 130.8, 129.7, 128.4, 124.5, 122.0, 113.0, 101.7, 57.7, 56.6, 49.6, 30.6, 30.3, 29.2, 22.8. MS m / z (ESI): 448.2 [M+1].

[0085] Single configuration compound 1-2: 1 H NMR (600 MHz, methanol-d4) δ 8.06 (s, 1H), 7.95 (d, J = 6.0 Hz, 1H), 7.61 (d, J = 8.7 Hz, 1H), 7.58-7.51 (m, 3H), 7.47-7.45 (m, 1H), 7.29 (dd, J = 8.7, 1.8 Hz, 1H), 4.34 (d, J = 13.3 Hz, 1H), 4.27 (d, J = 13.3 Hz, 1H), 4.02-3.98 (m, 1H), 3.69-3.64 (m, 1H), 2.32-2.27 (m, 1H), 2.15-2.10 (m, 1H), 2.03-1.96 (m, 1H), 1.90-1.74 (m, 5H). 13C NMR (300 MHz, methanol-d4) δ 159.9 (d, J = 15.9 Hz), 156.1, 141.0, 138.8, 138.7 (d, J = 244.2 Hz), 137.9, 134.2, 133.5, 131.5, 131.4 (d, J = 31.8 Hz), 131.2, 129.7, 128.5, 124.7, 121.7, 112.8, 107.9, 54.1, 53.7, 49.6, 31.8, 29.2, 28.8, 19.8. MS m / z (ESI): 448.2 [M+1].

[0086] Single configuration compounds 16-1 and 16-2 were prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0087] Single configuration compound 16-1: 1 H NMR (600 MHz, methanol-d4) δ 8.04 (s, 1H), 7.90 (d, J = 7.8 Hz, 1H), 7.63 (d, J = 8.2 Hz, 2H), 7.60 (d, J = 9.2 Hz, 1H), 7.49 (d, J = 8.2 Hz, 2H), 7.30-7.27 (m, 1H), 6.18 (d, J = 7.8 Hz, 1H), 4.34 (d, J = 13.3 Hz, 1H), 4.29 (d, J = 13.3 Hz, 1H), 4.03-3.99 (m, 1H), 3.70-3.65 (m, 1H), 2.36-2.31 (m, 1H), 2.14-2.10 (m, 1H), 2.03-1.96 (m, 1H), 1.90-1.76 (m, 5H). 13 C NMR (150 MHz, methanol-d4) δ 162.3, 150.2, 149.4, 140.5, 139.4, 134.5, 134.2, 133.7, 132.3, 128.3, 124.5, 121.9, 118.9, 117.0, 113.0, 110.1, 95.5, 54.8, 54.4, 49.6, 31.4, 28.9, 28.5, 19.6. MS m / z (ESI): 430.2 [M+1].

[0088] Single configuration compound 16-2: 1H NMR (600 MHz, methanol-d4) δ 8.21 (s, 1H), 7.75 (d, J = 7.8 Hz, 1H), 7.43 (d, J = 8.2 Hz, 2H), 7.31 (d, J = 9.2 Hz, 1H), 7.25 (d, J = 8.2 Hz, 2H), 7.22-7.19 (m, 1H), 6.15 (d, J = 7.8 Hz, 1H), 4.22 (d, J = 13.3 Hz, 1H), 4.15 (d, J = 13.3 Hz, 1H), 4.13-4.00 (m, 1H), 3.75-3.70 (m, 1H), 2.42-2.39 (m, 1H), 2.12-2.09 (m, 1H), 2.05-1.96 (m, 1H), 1.95-1.80 (m, 5H). 13 C NMR (150 MHz, methanol-d4) δ 165.1, 152.4, 148.4, 142.4, 138.2, 135.4, 134.4, 133.6, 132.4, 127.2, 123.4, 122.2, 119.0, 118.5, 114.1, 111.2, 96.4, 56.8, 55.4, 49.8, 32.2, 29.9, 28.7, 19.5. MS m / z (ESI): 430.2 [M+l].

[0089] Single configuration compounds 17-1 and 17-2 were prepared from cytosine 1j (Bide Pharmaceutics, CAS Number: 71-30-7).

[0090] Single configuration compound 17-1: 1 H NMR (600 MHz, methanol-d4) δ 7.75 (s, 1H), 7.49 (d, J = 6.9 Hz, 2H), 7.33 (d, J = 8.7 Hz, 1H), 7.11 (br s, 1H), 6.89 (dd, J = 8.7, 2.3 Hz, 1H), 6.51 (br s, 1H), 6.09 (d, J = 7.8 Hz, 1H), 4.45 (d, J = 11.9 Hz, 1H), 4.32 (br s, 1H), 3.86-3.82 (m, 1H), 3.57-3.36 (m, 4H), 2.17-2.10 (m, 1H), 2.10-2.02 (m, 2H), 1.97-1.91 (m, 1H), 1.88-1.79 (m, 2H). 13C NMR (150 MHz, methanol-d4) δ 162.0, 149.4, 148.8, 141.3, 140.2, 137.2, 133.6, 133.1, 132.0, 127.6, 124.7, 121.3, 118.9, 117.0, 112.4, 101.1, 95.4, 65.4, 61.7, 58.8, 53.9, 51.2, 34.6, 26.0, 22.9. MS m / z (ESI): 430.2 [M+1].

[0091] Single configuration compound 17-2: 1 H NMR (600 MHz, methanol-d4) δ 7.78 (s, 1H), 7.45 (d, J = 6.9 Hz, 2H), 7.37 (d, J = 8.7 Hz, 1H), 7.21 (br s, 1H), 6.65 (dd, J = 8.7, 2.3 Hz, 1H), 6.53 (br s, 1H), 6.19 (d, J = 7.8 Hz, 1H), 4.54 (d, J = 11.9 Hz, 1H), 4.37 (br s, 1H), 3.95 - 3.90 (m, 1H), 3.61 - 3.43 (m, 4H), 2.14 - 2.07 (m, 1H), 2.15 - 2.00 (m, 2H), 1.95 - 1.85 (m, 1H), 1.86 - 1.74 (m, 2H). 13 C NMR (150 MHz, methanol-d4) δ 162.4, 148.4, 148.1, 146.6, 142.1, 136.1, 132.3, 132.1, 131.0, 127.9, 124.5 121.3, 119.9, 117.4, 113.2, 103.2, 96.5, 67.9, 62.8, 59.2, 53.4, 52.1, 35.7, 27.1, 23.9. MS m / z (ESI): 430.2 [M+1].

[0092] Single configuration compounds 18-1 and 18-2 were prepared from cytosine 1j (Bide Pharm, CAS Number: 71-30-7).

[0093] Single configuration compound 18-1: 1H NMR (600 MHz, methanol-d4) δ 7.86 (s, 1H), 7.76 (d, J = 7.3 Hz, 1H), 7.55-7.50 (m, 3H), 7.46 (dt, J = 6.9, 1.8 Hz, 1H), 7.39 (d, J = 8.7 Hz, 1H), 7.03 (d, J = 8.7 Hz, 1H), 6.09 (d, J = 7.3 Hz, 1H), 4.30 (d, J = 13.3 Hz, 1H), 4.25 (d, J = 13.3 Hz, 1H), 3.94-3.91 (m, 1H), 3.57-3.52 (m, 1H), 2.45-2.41 (m, 1H), 2.20-2.16 (m, 1H), 1.90-1.74 (m, 5H), 1.65-1.59 (m, 1H). 13 C NMR (150 MHz, methanol-d4) δ 164.6, 153.0, 148.8, 141.0, 139.4.137.5, 133.9, 133.5, 131.6, 131.5, 129.4, 128.5, 125.0, 121.2, 112.2, 110.3, 96.0, 54.5, 50.7, 49.6, 32.7, 30.1, 29.5, 20.1. MS m / z (ESI): 430.2 [M+1].

[0094] Single configuration compound 18-2: 1 H NMR (600 MHz, methanol-d4) δ 7.83 (s, 1H), 7.74 (d, J = 7.3 Hz, 1H), 7.65-7.60 (m, 3H), 7.48 (dt, J = 6.9, 1.8 Hz, 1H), 7.33 (d, J = 8.7 Hz, 1H), 7.05 (d, J = 8.7 Hz, 1H), 6.03 (d, J = 7.3 Hz, 1H), 4.32 (d, J = 13.3 Hz, 1H), 4.25 (d, J = 13.3 Hz, 1H), 3.84-3.71 (m, 1H), 3.64-3.60 (m, 1H), 2.44-2.40 (m, 1H), 2.24-2.18 (m, 1H), 1.91-1.74 (m, 5H), 1.68-1.57 (m, 1H). 13C NMR (150 MHz, methanol-d4) δ 164.5, 153.8, 148.5, 141.5, 139.2 137.7 137.9, 134.5, 132.6, 135.5, 129.8, 128.9, 125.4, 121.5, 112.7, 112.3, 97.0, 51.5, 50.8, 49.5, 32.7, 30.1, 29.8, 20.8. MS m / z (ESI): 430.2 [M+1].

[0095] Single configuration compounds 19-1 and 19-2 were prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0096] Single configuration compound 19-1: 1 H NMR (600 MHz, methanol-d4) δ 7.84 (s, 1H), 7.69-7.63 (m, 2H), 7.58-7.48 (m, 2H), 7.38 (d, J = 9.2 Hz, 1H), 6.98 (dd, J = 8.9, 1.8 Hz, 1H), 6.90 (br s, 1H), 6.18 (d, J = 7.8 Hz, 1H), 4.46 (d, J = 13.3 Hz, 1H), 4.38 (d, J = 13.3 Hz, 1H), 3.82 (br s, 1H), 3.73-3.35 (m, 2H), 2.21-2.09 (m, 2H), 1.96-1.89 (m, 1H), 1.73-1.51 (m, 1H). 13 C NMR (150 MHz, methanol-d4) δ 162.1, 150.2, 149.0, 141.1, 141.0, 140.9, 137.4, 133.5, 133.3, 131.8, 128.4, 124.9, 121.2, 118.6, 112.4, 101.9, 95.5, 67.5, 60.7, 55.9, 55.2, 29.1, 22.3. MS m / z (ESI): 416.2 [M+1].

[0097] Single configuration compound 19-2: 1H NMR (600 MHz, methanol-d4) δ 7.86 (s, 1H), 7.75-7.70 (m, 2H), 7.59-7.53 (m, 2H), 7.35 (d, J = 9.2 Hz, 1H), 6.88 (dd, J = 8.9, 1.8 Hz, 1H), 6.92 (br s, 1H), 6.51 (d, J = 7.8 Hz, 1H), 4.42 (d, J = 13.3 Hz, 1H), 4.32 (d, J = 13.3 Hz, 1H), 3.85 (br s, 1H), 3.77-3.41 (m, 2H), 2.31-2.02 (m, 2H), 1.95-1.85 (m, 1H), 1.73-1.54 (m, 1H).

[0098] 13 C NMR (150 MHz, methanol-d4) δ 162.1, 153.1, 149.3, 142.3, 141.2, 141.0, 138.4, 134.2, 133.5, 131.7, 126.2, 124.1, 122.3, 119.3, 112.3, 102.3, 95.5, 68.7, 61.4, 56.3, 55.3, 29.2, 22.3. MS m / z (ESI): 416.2 [M+l].

[0099] Single configuration compound 20-1 was prepared from cytosine 1j (Bide Pharmaceutics, CAS Number: 71-30-7).

[0100] Single configuration compound 20-1: 1 H NMR (600 MHz, DMSO-d6) δ 9.59 (s, 1H), 8.56 (d, J = 6.9 Hz, 1H), 8.43 (d, J = 6.9 Hz, 1H), 7.90 (d, J = 7.3 Hz, 1H), 7.87 (d, J = 7.8 Hz, 1H), 7.75 (t, J = 8.2 Hz, 1H), 7.71 (d, J = 7.8 Hz, 1H), 7.47 (t, J = 8.2 Hz, 1H), 7.46-7.44 (m, 2H), 7.35 (dd, J = 7.3, 1.8 Hz, 1H), 7.08 (s, 1H), 6.40 (s, 1H), 6.14 (d, J = 7.8 Hz, 1H), 4.38 (s, 2H). 13C NMR (150 MHz, DMSO-d6) δ 165.5, 152.0, 148.5, 141.8, 138.5, 136.6, 134.2, 132.5, 130.4, 129.4, 128.7, 128.4, 126.1, 124.9, 122.5, 119.3, 119.2, 117.6, 109.2, 103.9, 103.2, 102.7, 91.5, 48.0. MS m / z (ESI): 435.2 [M+1].

[0101] Single configuration compound 21-1 was prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0102] Single configuration compound 21-1: 1 H NMR (600 MHz, DMSO-d6) δ 9.59 (s, 1H), 8.56 (d, J = 6.9 Hz, 1H), 8.43 (d, J = 6.9 Hz, 1H), 7.90 (d, J = 7.8 Hz, 1H), 7.87 (d, J = 7.8 Hz, 1H), 7.75 (t, J = 8.2 Hz, 1H), 7.71 (d, J = 7.8 Hz, 1H), 7.46 - 7.44 (m, 2H), 7.35 (dd, J = 7.3, 1.8 Hz, 1H), 7.08 (s, 1H), 6.40 (s, 1H), 6.14 (d, J = 7.8 Hz, 1H), 4.38 (s, 2H). 13 C NMR (150 MHz, DMSO-d6) δ 166.5, 153.0, 147.5, 142.8, 134.2, 135.1, 134.4, 133.5, 124.1, 127.1, 127.0, 125.1, 124.9, 122.5, 119.3, 119.2, 117.6, 109.2, 107.3, 103.9, 103.2, 91.5, 48.0. MS m / z (ESI): 425.2 [M+1].

[0103] Single configuration compound 22-1 was prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0104] Single configuration compound 22-1: 1H NMR (600 MHz, methanol-d4) δ 7.82 (s, 1H), 7.75 (d, J = 7.3 Hz, 1H), 7.66-7.55 (m, 2H), 7.49 (s, 1H), 7.34 (d, J = 8.7 Hz, 1H), 7.04 (d, J = 8.7 Hz, 1H), 6.03 (d, J = 7.3 Hz, 1H), 4.32 (d, J = 13.3 Hz, 1H), 4.25 (d, J = 13.3 Hz, 1H), 3.84-3.71 (m, 1H), 3.65-3.60 (m, 1H), 2.44-2.40 (m, 1H), 2.24-2.18 (m, 1H), 1.91-1.74 (m, 5H), 1.68-1.57 (m, 1H). 13 C NMR (150 MHz, methanol-d4) δ 164.4, 153.1, 148.5, 141.6, 139.7 137.8 137.2, 134.4, 132.5, 135.5, 129.8, 128.3, 125.6, 121.7, 112.7, 112.3, 96.0, 51.7, 50.8, 49.8, 32.7, 30.1, 29.8, 20.8. MS m / z (ESI): 464.2 [M+l].

[0105] Single configuration compound 23-1 was prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0106] Single configuration compound 23-1: 1 H NMR (600 MHz, methanol-d4) δ 7.83 (s, 1H), 7.75 (d, J = 7.3 Hz, 1H), 7.66-7.53 (m, 2H), 7.49 (s, 1H), 7.39 (d, J = 8.7 Hz, 1H), 7.09 (d, J = 8.7 Hz, 1H), 6.04 (d, J = 7.3 Hz, 1H), 4.37 (d, J = 13.3 Hz, 1H), 4.28 (d, J = 13.3 Hz, 1H), 3.85-3.79 (m, 1H), 3.69-3.69 (m, 1H), 2.47-2.40 (m, 1H), 2.24-2.18 (m, 1H), 1.91-1.74 (m, 5H), 1.68-1.57 (m, 1H). 13C NMR (150 MHz, methanol-d4) δ 167.4, 155.1, 147.9, 144.3, 138.4 137.3 137.1, 134.4, 132.5, 135.9, 129.1, 128.6, 126.7, 123.2, 112.2, 113.2, 96.1, 51.7, 50.8, 49.8, 32.7, 30.1, 29.8, 20.8. MS m / z (ESI): 446.2 [M+1].

[0107] Single configuration compound 24-1 was prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0108] Single configuration compound 24-1: 1 H NMR (600 MHz, methanol-d4) δ 8.12 (s, 1H), 7.78 (d, J = 7.8 Hz, 1H), 7.72 - 7.64 (m, 2H), 7.14 (s, 1H), 6.91 (d, J = 8.5 Hz, 1H), 6.62 (d, J = 8.5 Hz, 1H), 6.14 (d, J = 7.8 Hz, 1H), 4.39 (d, J = 12.0 Hz, 1H), 4.38 (d, J = 12.0 Hz, 1H), 3.95 - 3.79 (m, 1H), 3.69 - 3.69 (m, 1H), 2.47 - 2.40 (m, 1H), 2.34 - 2.28 (m, 1H), 1.91 - 1.74 (m, 5H), 1.68 - 1.57 (m, 1H). 13 C NMR (150 MHz, methanol-d4) δ 167.4, 155.1, 147.9, 144.3, 138.4 137.3 137.1, 134.4, 132.5, 135.9, 129.1, 128.6, 126.7, 123.2, 112.2, 113.2, 96.1, 51.7, 50.8, 49.8, 32.7, 30.1, 29.8, 20.8. MS m / z (ESI): 446.2 [M+1].

[0109] Single configuration compound 25-1 was prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0110] Single configuration compound 25-1: 1H NMR (600 MHz, methanol-d4) δ 8.20 (s, 1H), 7.52 (d, J = 7.8 Hz, 1H), 7.70-7.63 (m, 2H), 7.10 (s, 1H), 6.81 (d, J = 8.7 Hz, 1H), 6.32 (d, J = 8.7 Hz, 1H), 6.04 (d, J = 7.3 Hz, 1H), 4.38 (d, J = 13.3 Hz, 1H), 4.28 (d, J = 13.3 Hz, 1H), 3.85-3.79 (m, 1H), 3.81 (s, 3H), 3.69-3.62 (m, 1H), 2.47-2.40 (m, 1H), 2.24-2.18 (m, 1H), 1.91-1.74 (m, 5H), 1.68-1.57 (m, 1H). 13 C NMR (150 MHz, methanol-d4) δ 166.4, 154.1, 147.5, 142.6, 139.5 137.7 137.3, 134.3, 132.5, 131.8, 129.8, 128.4, 125.7, 121.2, 112.1, 112.2, 96.0, 55.8, 51.7, 50.8, 49.8, 32.7, 30.1, 29.8, 20.8. MS m / z (ESI): 460.2 [M+l].

[0111] Single configuration compound 26-1 was prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0112] Single configuration compound 26-1: 1 H NMR (600 MHz, methanol-d4) δ 8.32 (s, 1H), 8.12 (d, J = 7.8 Hz, 1H), 7.87-7.72 (m, 2H), 7.52 (s, 1H), 6.98 (d, J = 8.7 Hz, 1H), 6.45 (d, J = 8.7 Hz, 1H), 6.21 (d, J = 7.3 Hz, 1H), 4.39 (d, J = 13.3 Hz, 1H), 4.32 (d, J = 13.3 Hz, 1H), 3.86-3.82 (m, 1H), 3.68-3.61 (m, 1H), 2.47-2.40 (m, 1H), 2.24-2.18 (m, 1H), 1.91-1.74 (m, 5H), 1.68-1.57 (m, 1H). 13C NMR (150 MHz, methanol-d4) δ 168.2, 155.2, 145.5, 144.6, 137.5 135.7 133.3, 132.3, 131.5, 130.8, 129.8, 128.4, 125.7, 121.2, 118.6, 112.2, 112.2, 96.0, 51.2, 50.3, 49.4, 32.5, 30.1, 29.7, 20.8. MS m / z (ESI): 445.2 [M + 1].

[0113] Single configuration compound 27-1 was prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0114] Single configuration compound 27-1: 1 H NMR (600 MHz, methanol-d4) δ 8.15 (s, 1H), 7.92 (d, J = 7.8 Hz, 1H), 7.82 - 7.810 (m, 2H), 7.75 (s, 1H), 7.42 (d, J = 8.5 Hz, 1H), 6.63 (d, J = 8.5 Hz, 1H), 6.54 (d, J = 7.8 Hz, 1H), 4.33 (d, J = 12.0 Hz, 1H), 4.30 (d, J = 12.0 Hz, 1H), 4.06 (s, 3H), 3.92 - 3.72 (m, 1H), 3.61 - 3.52 (m, 1H), 2.42 - 2.35 (m, 1H), 2.32 - 2.28 (m, 1H), 1.91 - 1.74 (m, 5H), 1.68 - 1.57 (m, 1H). 13 C NMR (150 MHz, methanol-d4) δ 173.2, 169.2, 159.1, 148.9, 148.3, 141.4 138.2 137.2, 132.4, 131.4, 130.2, 128.1, 127.1, 125.2, 123.8, 112.7, 110.2, 92.1, 51.1, 50.7, 49.2, 32.4, 30.3, 29.2, 20.1. MS m / z (ESI): 474.2 [M + 1].

[0115] Single configuration compound 28-1 was prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0116] Single configuration compound 28-1: 1H NMR (600 MHz, methanol-d4) δ 7.82 (d, J = 7.5 Hz, 1H), 7.57-7.52 (m, 3H), 7.50 (dt, J = 6.9, 1.8 Hz, 1H), 7.35 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 8.5 Hz, 1H), 6.59 (d, J = 7.5 Hz, 1H), 4.33 (d, J = 13.1 Hz, 1H), 4.23 (d, J = 13.1 Hz, 1H), 3.96-3.93 (m, 1H), 3.62-3.54 (m, 1H), 2.44-2.42 (m, 1H), 2.22-2.15 (m, 1H), 1.94-1.73 (m, 5H), 1.63-1.51 (m, 1H). 13 C NMR (150 MHz, methanol-d4) δ 168.3, 150.0, 145.8, 140.5, 137.2. 137.0, 134.2, 133.1, 131.2, 131.0, 129.2, 128.4, 125.5, 122.4, 112.5, 110.5, 96.4, 54.7, 54.0, 50.8, 49.5, 32.2, 32.1, 28.5, 21.1. MS m / z (ESI): 460.2 [M+1].

[0117] Single configuration compound 29-1 was prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0118] Single configuration compound 29-1: 1 H NMR (600 MHz, methanol-d4) δ 7.72 (d, J = 7.3 Hz, 1H), 7.54-7.49 (m, 3H), 7.42 (dt, J = 6.9, 1.8 Hz, 1H), 7.35 (d, J = 8.7 Hz, 1H), 7.04 (d, J = 8.7 Hz, 1H), 6.08 (d, J = 7.3 Hz, 1H), 4.35 (d, J = 13.3 Hz, 1H), 4.29 (d, J = 13.3 Hz, 1H), 3.97-3.90 (m, 1H), 3.58-3.51 (m, 1H), 2.44-2.40 (m, 1H), 2.25-2.14 (m, 1H), 2.06 (s, 3H), 1.91-1.84 (m, 5H), 1.67-1.59 (m, 1H). 13C NMR (150 MHz, methanol-d4) δ 167.6, 152.0, 148.0, 142.1, 138.1, 137.2, 133.9, 133.4, 131.2, 131.1, 129.2, 128.5, 125.0, 121.2, 112.2, 110.3, 96.0, 54.5, 52.7, 48.6, 32.7, 30.8, 27.5, 20.1, 14.1. MS m / z (ESI): 444.2 [M+1].

[0119] Single configuration compound 30-1 was prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0120] Single configuration compound 30-1: 1 H NMR (600 MHz, methanol-d4) δ 7.86 (d, J = 7.3 Hz, 1H), 7.52 - 7.45 (m, 3H), 7.41 (dt, J = 6.9, 1.8 Hz, 1H), 7.39 (d, J = 8.7 Hz, 1H), 7.23 (d, J = 8.7 Hz, 1H), 6.49 (d, J = 7.3 Hz, 1H), 4.32 (d, J = 12.0 Hz, 1H), 4.25 (d, J = 12.0 Hz, 1H), 3.92 - 3.85 (m, 1H), 3.57 - 3.52 (m, 1H), 2.45 - 2.41 (m, 1H), 2.20 - 2.16 (m, 1H), 1.90 - 1.74 (m, 5H), 1.65 - 1.59 (m, 1H). 13 C NMR (150 MHz, methanol-d4) δ 164.6, 153.0, 148.8, 141.0, 139.4. 137.5, 133.9, 133.5, 131.6, 131.5, 129.4, 128.5, 125.0, 121.2, 112.2, 110.3, 96.0, 54.5, 50.7, 49.6, 32.7, 30.1, 29.5, 20.1. MS m / z (ESI): 508.1 [M+1].

[0121] Single configuration compound 31-1 was prepared from cytosine 1j (Biotrend, CAS Number: 71-30-7).

[0122] Single configuration compound 31-1: 1H NMR (600 MHz, methanol-d4) δ 7.96 (d, J = 7.8 Hz, 1H), 7.57-7.51 (m, 3H), 7.39 (dt, J = 6.9, 1.8 Hz, 1H), 7.35 (d, J = 8.7 Hz, 1H), 7.23 (d, J = 8.7 Hz, 1H), 6.19 (d, J = 7.3 Hz, 1H), 4.30 (d, J = 13.3 Hz, 1H), 4.25 (d, J = 13.3 Hz, 1H), 3.94-3.91 (m, 1H), 3.57-3.52 (m, 1H), 2.45-2.41 (m, 1H), 2.20-2.16 (m, 1H), 1.90-1.74 (m, 5H), 1.65-1.59 (m, 1H). 13 C NMR (150 MHz, methanol-d4) δ 169.6, 154.2, 148.4, 141.0, 139.4.137.5, 133.9, 133.5, 131.6, 131.5, 129.4, 128.5, 125.0, 121.2, 112.7, 112.2, 110.3, 96.0, 54.5, 50.7, 49.6, 32.7, 30.1, 29.5, 20.1. MS m / z (ESI): 445.2 [M+1].

[0123] Example 3

[0124] The compounds in Example 3 are all obtained from the following reaction general formula:

[0125]

[0126] First step, synthesis of 1i:

[0127] The compound 1c with different substituents (2.36 mmol), 3-formylphenylboronic acid 1h (Bide Pharmatech, CAS No.: 87199-16-4) (708 mg, 4.72 mmol), copper acetate (944 mg, 4.72 mmol), pyridine (4.5 mL) were added into dichloromethane (120 mL) and reacted at room temperature for 12 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with dichloromethane (500 mL) containing 1% volume percent of triethylamine to obtain the corresponding compound 1i with a yield of 73-92%.

[0128] Sixth step, synthesis of II-1 and II-2:

[0129] The corresponding compound 1i (0.43 mmol), compound 1g (100 mg, 0.43 mmol), sodium cyanoborohydride (48 mg, 0.76 mmol) were added to anhydrous methanol (20 mL), and the pH was adjusted to 6.0-7.0 with acetic acid, and the reaction was carried out at room temperature for 12 hours. Water (1 mL) was added, and the solvent was removed under reduced pressure, and the residue was purified by high performance liquid chromatography (separation conditions: preparative column Welch XB-C18, 21.2 x 250 mm, 5 μm; elution system: methanol / water (volume ratio) = 10%-100% containing 0.5% volume fraction of trifluoroacetic acid, flow rate: 10 ml / min, elution time: 40 min) to obtain the corresponding compounds II-1 and II-2 with a yield of 23-32% and 54-63%, respectively.

[0130] The single configuration compounds 2-1 and 2-2 were prepared from 4-aminopyridin-2-one 1c (Bide Pharm, CAS No.: 38767-72-5).

[0131] Single configuration compound 2-1: 1 H NMR (600 MHz, methanol-d4) δ 8.14 (s, 1H), 7.71 (dd, J = 8.9, 1.4 Hz, 1H), 7.54 (t, J = 7.8 Hz, 1H), 7.50 (d, J = 8.2 Hz, 1H), 7.47 (t, 1.4 Hz, 1H), 7.41 (d, J = 8.7 Hz, 1H), 7.38-7.36 (m, 1H), 7.36 (d, J = 7.8 Hz, 1H), 6.13 (d, J = 7.3 Hz, 1H), 4.31 (d, J = 13.3 Hz, 1H), 4.24 (d, J = 13.3 Hz, 1H), 3.70-3.65 (m, 1H), 3.32-3.27 (m, 1H), 2.44-2.41 (m, 1H), 2.29-2.25 (m, 1H), 2.13-2.10 (m, 1H), 2.08-2.05 (m, 1H), 1.65 (q, J = 11.9 Hz, 1H), 1.54-1.42 (m, 3H). 13 C NMR (300 MHz, methanol-d4) δ 164.8, 161.1, 142.4, 141.9, 140.9, 139.5, 135.2, 133.4, 131.4, 130.9, 130.0, 128.9, 124.2, 122.4, 118.7, 113.4, 107.1, 103.0, 59.7, 56.3, 49.6, 33.2, 29.6, 29.0, 22.5. MS m / z (ESI): 429.2 [M+1].

[0132] Single configuration compound 2-2: 1 H NMR (600 MHz, methanol-d4) δ 7.84 (s, 1H), 7.49 (t, J = 7.8 Hz, 1H), 7.44-7.42 (m, 2H), 7.38-7.65 (m, 2H), 7.26 (d, J = 7.3 Hz, 1H), 7.00 (d, J = 8.7 Hz, 1H), 6.02 (d, J = 7.3 Hz, 1H), 4.26 (d, J = 13.3 Hz, 1H), 4.21 (d, J = 13.3 Hz, 1H), 3.93-3.91 (m, 1H), 3.66-3.50 (m, 1H), 2.45-2.42 (m, 1H), 2.20-2.16 (m, 1H), 1.90-1.70 (m, 5H), 1.61-1.54 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 165.5, 160.7, 142.7, 142.1, 141.9, 139.2, 133.6, 133.4, 131.4, 130.8, 130.0, 128.9, 124.9, 121.4, 121.2, 112.2, 108.2, 102.5, 54.5, 50.1, 49.6, 32.9, 30.1, 29.6, 20.1. MS m / z (ESI): 429.2 [M+1].

[0133] Single configuration compounds 3-1 and 3-2 were prepared from 4-methyl-5-nitropyridin-2-one 1c (Bide Pharm, CAS Number: 21901-41-7).

[0134] Single configuration compound 3-1: 1 H NMR (600 MHz, methanol-d4) δ 7.95 (s, 1H), 7.46 (d, J = 8.7 Hz, 1H), 7.26 (t, J = 7.6 Hz, 2H), 7.05 (d, J = 8.7 Hz, 1H), 6.90-6.81 (m, 5H), 4.48-4.02 (m, 2H), 3.38-3.33 (m, 1H), 3.34 (s, 3H), 3.30-3.26 (m, 1H), 2.55-2.50 (m, 1H), 2.20-2.15 (m, 1H), 2.11-2.03 (m, 2H), 1.61 (q, J = 11.5 Hz, 1H), 1.45-1.27 (m, 3H). 13C NMR (300 MHz, methanol-d4) δ 159.5, 159.4, 144.2, 138.2, 133.9, 133.9, 131.6, 130.4, 124.9, 122.7, 121.4, 119.1, 118.8, 117.9, 115.1, 114.8, 112.5, 109.9, 65.1, 55.5, 55.4, 49.6, 33.0, 32.2, 26.9, 23.6. MS m / z (ESI): 443.6 [M+1].

[0135] Single configuration compound 3-2: 1 H NMR (600 MHz, methanol-d4) δ 7.81 (s, 1H), 7.34 (d, J = 9.2 Hz, 1H), 7.27-7.22 (m, 1H), 6.91-6.76 (m, 5H), 6.65 (s, 1H), 6.52 (d, J = 5.5 Hz, H), 4.42 (d, J = 12.8 Hz, 1H), 4.24 (d, J = 12.8 Hz, 1H), 4.09-4.02 (m, 1H), 3.71-3.64 (m, 1H), 3.33 (s, 3H), 2.62-2.56 (m, 1H), 2.18-2.14 (m, 1H), 1.91-1.71 (m, 6H). 13 C NMR (300 MHz, methanol-d4) δ 159.4, 159.2, 142.9, 142.9, 136.9, 133.2, 132.3, 131.6, 131.4, 125.1, 123.4, 123.0, 121.1, 119.0, 118.6, 118.0, 117.8, 112.1, 59.4, 55.3, 55.2, 49.6, 30.8, 29.1, 27.6, 21.1. MS m / z (ESI): 443.6 [M+1].

[0136] Single configuration compounds 4-1 and 4-2 were prepared from 2-hydroxy-4- methylpyridine 1c (Bide Pharm, CAS number: 13466-41-6).

[0137] Single configuration compound 4-1: 1H NMR (600 MHz, methanol-d4) δ 8.00 (s, 1H), 7.60 (t, J = 7.3 Hz, 1H), 7.57-7.54 (m, 2H), 7.51 (s, 1H), 7.49-7.47 (m, 2H), 7.42 (d, J = 8.2 Hz, 1H), 7.23 (d, J = 8.2 Hz, 1H), 6.48 (s, 1H), 6.40 (d, J = 7.3 Hz, 1H), 4.32 (d, J = 12.8 Hz, 1H), 4.26 (d, J = 12.8 Hz, 1H), 3.60-3.55 (m, 1H), 3.34-3.29 (m, 1H), 2.52-2.48 (m, 1H), 2.30 (s, 3H), 2.28-2.24 (m, 1H), 2.14-2.10 (m, 1H), 2.06-2.02 (m, 1H), 1.55-1.39 (m, 4H). 13 CNMR (300 MHz, methanol-d4) δ 164.4, 155.8, 142.4, 139.4, 139.4, 138.5, 134.4, 133.8, 131.6, 131.3, 129.6, 128.8, 124.6, 121.9, 119.8, 112.8, 111.4, 110.2, 56.9, 56.8, 49.6, 34.5, 30.9, 29.3, 22.9, 21.4. MS m / z (ESI): 428.2 [M+1].

[0138] Single configuration compound 4-2: 1 H NMR (600 MHz, methanol-d4) δ 7.93 (s, 1H), 7.56 (t, J = 7.3 Hz, 1H), 7.54-7.51 (m, 2H), 7.48-7.42 (m, 3H), 7.23-7.20 (m, 1H), 7.14-7.11 (m, 1H), 6.46 (s, 1H), 6.37 (dd, J = 6.4, 1.8 Hz, 1H), 4.32 (d, J = 12.8 Hz, 1H), 4.26 (d, J = 12.8 Hz, 1H), 3.98-3.94 (m, 1H), 3.61-3.57 (m, 1H), 2.41-2.36 (m, 1H), 2.29 (s, 3H), 2.19-2.14 (m, 1H), 1.88-1.78 (m, 5H), 1.69-1.61 (m, 1H). 13C NMR (300 MHz, methanol-d4) δ 164.4, 155.8, 142.4, 138.5, 138.5, 138.3, 133.9, 133.7, 131.6, 131.4, 129.7, 128.8, 124.8, 121.5, 119.8, 112.6, 111.4, 110.0, 54.3, 52.6, 49.6, 32.2, 29.5, 29.1, 21.4, 19.9. MS m / z (ESI): 428.2 [M+1].

[0139] Single configuration compounds 5-1 and 5-2 were prepared from 2-pyridone 1c (Bide Pharm, CAS Number: 142-08-5).

[0140] Single configuration compound 5-1: 1 H NMR (600 MHz, methanol-d4) δ 8.11 (s, 1H), 7.69-7.63 (m, 2H), 7.61-7.58 (m, 2H), 7.57-7.53 (m, 2H), 7.44-7.42 (m, 1H), 7.37 (d, J = 8.7 Hz, 1H), 6.66 (d, J = 9.2 Hz, 1H), 6.51 (td, J = 6.9, 1.4 Hz, 1H), 4.34 (d, J = 12.8 Hz, 1H), 4.26 (d, J = 12.8 Hz, 1H), 3.69-3.63 (m, 1H), 3.34-3.29 (m, 1H), 2.47-2.42 (m, 1H), 2.29-2.25 (m, 1H), 2.14-2.11 (m, 1H), 2.08-2.05 (m, 1H), 1.63 (q, J = 11.9 Hz, 1H), 1.52-1.44 (m, 3H). 13 C NMR (300 MHz, methanol-d4) δ 164.5, 143.2, 142.5, 140.5, 139.8, 139.7, 135.0, 133.8, 131.7, 131.4, 129.6, 128.8, 124.3, 122.2, 121.6, 113.2, 111.1, 108.9, 59.0, 56.4, 49.6, 33.5, 29.9, 29.0, 22.6. MS m / z (ESI): 414.2 [M+1].

[0141] Single configuration compound 5-2: 1H NMR (600 MHz, methanol-d4) δ 7.92 (s, 1H), 7.66-7.62 (m, 1H), 7.59-7.52 (m, 4H), 7.48-7.44 (m, 2H), 7.22-7.19 (m, 1H), 7.14-7.10 (m, 1H), 6.65 (d, J = 9.2 Hz, 1H), 6.48 (td, J = 7.3, 1.8 Hz, 1H), 4.32 (d, J = 12.8 Hz, 1H), 4.27 (d, J = 12.8 Hz, 1H), 3.94-3.98 (m, 1H), 3.57-3.62 (m, 1H), 2.42-2.37 (m, 1H), 2.20-2.14 (m, 1H), 1.89-1.78 (m, 5H), 1.70-1.63 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 164.5, 143.2, 142.5, 139.7, 138.3, 138.2, 133.9, 133.8, 131.6, 131.5, 129.6, 128.8, 124.8, 121.6, 121.5, 112.5, 108.9, 105.7, 54.3, 52.4, 49.6, 30.2, 29.5, 29.1, 19.9. MS m / z (ESI): 414.2 [M+l].

[0142] Single configuration compounds 6-1 and 6-2 were prepared from 2-oxo-1,2- dihydropyridine-4-carbonitrile 1c (Bide Pharm, CAS Number: 94805-51-3).

[0143] Single configuration compound 6-1: 1 H NMR (600 MHz, methanol-d4) δ 8.14 (s, 1H), 7.76 (d, J = 6.9 Hz, 1H), 7.71 (dd, J = 9.2, 0.9 Hz, 1H), 7.61-7.55 (m, 3H), 7.45 (dt, J = 6.9, 1.8 Hz, 1H), 7.42 (d, J = 9.2 Hz, 1H), 7.06 (d, J = 1.8 Hz, 1H), 6.62 (dd, J = 6.9, 1.8 Hz, 1H), 4.34 (d, J = 13.3 Hz, 1H), 4.26 (d, J = 13.3 Hz, 1H), 3.70-3.64 (m, 1H), 3.33-3.29 (m, 1H), 2.48-2.43 (m, 1H), 2.29-2.25 (m, 1H), 2.13-2.09 (m, 1H), 2.08-2.04 (m, 1H), 1.69 (q, J = 11.9 Hz, 1H), 1.56-1.45 (m, 3H).13 C NMR (300 MHz, methanol-d4) δ 162.4, 141.8, 141.7, 141.0, 140.8, 135.2, 134.0, 131.8, 131.7, 129.3, 128.6, 127.7, 126.5, 124.2, 122.4, 116.6, 115.9, 113.3, 107.6, 59.7, 56.4, 49.6, 33.2, 29.5, 28.9, 22.5. MS m / z (ESI): 414.2 [M+1].

[0144] Single configuration compound 6-2: 1 H NMR (600 MHz, methanol-d4) δ 7.84 (s, 1H), 7.70 (d, J = 6.9 Hz, 1H), 7.58-7.52 (m, 3H), 7.46 (dt, J = 7.3, 1.8 Hz, 1H), 7.37 (d, J = 9.6 Hz, 1H), 7.05 (d, J = 1.4 Hz, 1H), 7.00 (d, J = 9.2 Hz, 1H), 6.57 (dd, J = 6.9, 1.8 Hz, 1H), 4.30 (d, J = 13.3 Hz, 1H), 4.26 (d, J = 13.3 Hz, 1H), 3.93-3.90 (m, 1H), 3.55-3.50 (m, 1H), 2.47-2.42 (m, 1H), 2.21-2.16 (m, 1H), 1.89-1.71 (m, 5H), 1.62-1.55 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 162.4, 141.8, 141.7, 141.0, 140.8, 135.2, 134.0, 131.8, 131.7, 129.3, 128.6, 127.7, 126.5, 124.2, 122.4, 116.6, 115.9, 113.3, 107.6, 59.7, 56.4, 49.6, 33.2, 29.5, 28.9, 22.5. MS m / z (ESI): 414.2 [M+1].

[0145] Single configuration compounds 7-1 and 7-2 were prepared from 2-oxo-1,2-dihydropyridine-4-carboxamide 1c (Bide Pharm, CAS number: 175277-69-7).

[0146] Single configuration compound 7-1: 1H NMR (600 MHz, methanol-d4) δ 7.92 (s, 1H), 7.69 (d, J = 6.9 Hz, 1H), 7.64 (t, J = 7.8 Hz, 1H), 7.60-7.58 (m, 1H), 7.56 (t, J = 1.4 Hz, 1H), 7.49-7.47 (m, 2H), 7.11 (d, J = 8.7 Hz, 1H), 7.05 (d, J = 1.4 Hz, 1H), 6.81 (dd, J = 7.3, 1.8 Hz, 1H), 4.34 (d, J = 13.3 Hz, 1H), 4.28 (d, J = 13.3 Hz, 1H), 3.55-3.50 (m, 1H), 3.37-3.32 (m, 1H), 2.56-2.52 (m, 1H), 2.29-2.25 (m, 1H), 2.16-2.11 (m, 1H), 2.06-2.01 (m, 1H), 1.57-1.49 (m, 1H), 1.47-1.38 (m, 2H), 1.38-1.31 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 169.0, 164.3, 147.8, 142.2, 140.2, 140.0, 137.8, 134.8, 133.8, 131.8, 131.6, 129.5, 128.7, 124.4, 122.0, 120.5, 113.1, 106.6, 106.5, 58.0, 56.6, 49.6, 34.0, 30.4, 29.2, 22.7. MS m / z (ESI): 457.2 [M+1].

[0147] Single configuration compound 7-2: 1 H NMR (600 MHz, methanol-d4) δ 7.90 (s, 1H), 7.65 (d, J = 6.9 Hz, 1H), 7.58 (t, J = 8.2 Hz, 1H), 7.56-7.53 (m, 2H), 7.47 (dt, J = 7.8, 1.4 Hz, 1H), 7.4 (d, J = 8.7 Hz, 1H), 7.09 (d, J = 9.2 Hz, 1H), 7.04 (d, J = 1.4 Hz, 1H), 6.78 (dd, J = 6.9, 1.8 Hz, 1H), 4.32 (d, J = 13.3 Hz, 1H), 4.26 (d, J = 13.3 Hz, 1H), 3.96-3.93 (m, 1H), 3.60-3.55 (m, 1H), 2.43-2.38 (m, 1H), 2.22-2.15 (m, 1H), 1.88-1.77 (m, 5H), 1.67-1.61 (m, 1H). 13C NMR (300 MHz, methanol-d4) δ 169.0, 164.3, 147.8, 142.2, 140.2, 139.3, 138.2, 134.4, 133.7, 131.8, 131.7, 129.6, 128.8, 124.6, 121.8, 120.5, 113.0, 106.5, 105.1, 54.7, 54.2, 49.6, 31.6, 28.9, 28.6, 19.7. MS m / z (ESI): 457.2 [M + 1].

[0148] Single configuration compounds 8-1 and 8-2 were prepared from 2-hydroxy-3- methylpyridine 1c (Bide Pharm, CAS Number: 1003-56-1).

[0149] Single configuration compound 8-1: 1 H NMR (600 MHz, methanol-d4) δ 7.98 (s, 1H), 7.60 (t, J = 7.8 Hz, 1H), 7.57-7.54 (m, 2H), 7.53-7.51 (m, 2H), 7.46-7.42 (m, 2H), 7.22 (d, J = 9.2 Hz, 1H), 6.42 (t, J = 6.9 Hz, 1H), 4.33 (d, J = 13.3 Hz, 1H), 4.26 (d, J = 13.3 Hz, 1H), 3.60-3.54 (m, 1H), 3.35-3.30 (m, 1H), 2.53-2.49 (m, 1H), 2.28-2.24 (m, 1H), 2.15 (s, 3H), 2.15-2.10 (m, 1H), 2.06-2.01 (m, 1H), 1.54-1.39 (m, 4H). 13 C NMR (300 MHz, methanol-d4) δ 164.6, 143.0, 140.6, 140.2, 137.0, 135.0, 133.7, 131.6, 131.2, 131.1, 129.7, 128.8, 124.3, 122.3, 118.9, 117.0, 113.2, 108.5, 59.1, 56.4, 49.6, 33.5, 29.9, 29.0, 22.6, 17.2. MS m / z (ESI): 428.2 [M + 1].

[0150] Single configuration compound 8-2: 1H NMR (600 MHz, methanol-d4) δ 7.87 (d, J = 0.9 Hz, 1H), 7.55 (t, J = 7.8 Hz, 1H), 3.52-3.49 (m, 3H), 7.44-7.42 (m, 1H), 7.42-7.39 (m, 2H), 7.05 (d, J = 8.7 Hz, 1H), 6.38 (t, J = 6.9 Hz, 1H), 4.30 (d, J = 13.3 Hz, 1H), 4.26 (d, J = 13.3 Hz, 1H), 3.95-3.92 (m, 1H), 3.58-3.52 (m, 1H), 2.45-2.40 (m, 1H), 2.20-2.15 (m, 1H), 2.14 (s, 3H), 1.90-1.75 (m, 5H), 1.65-1.58 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 164.6, 143.0, 140.2, 140.2, 139.2, 137.0, 134.3, 133.5, 131.6, 131.4, 131.1, 129.8, 128.9, 124.6, 121.8, 118.8, 112.9, 108.5, 54.5, 54.0, 49.6, 31.7, 28.9, 28.7, 19.7, 17.2. MS m / z (ESI): 428.2 [M+1].

[0151] Single configuration compounds 9-1 and 9-2 were prepared from 5-methylpyridin-2-one 1c (Bide Pharm, CAS Number: 1003-68-5).

[0152] Single configuration compound 9-1: 1H NMR (600 MHz, methanol-d4) δ 8.09 (d, J = 1.2 Hz, 1H), 7.72 (br, 1H), 7.66 (d, J = 8.7 Hz, 1H), 7.59 (t, J = 7.78 Hz, 1H), 7.56-7.54 (m, 2H), 7.51 (t, J = 1.83, 1H), 7.43-7.41 (m, 1H), 7.40 (s, 1H), 7.35 (d, J = 8.7 Hz, 1H), 6.60 (d, J = 9.6 Hz, 1H), 4.33 (d, J = 12.8 Hz, 1H), 4.26 (d, J = 13.3 Hz, 1H), 3.66-3.62 (m, 1H), 3.33-3.29 (m, 1H), 2.47-2.45 (m, 1H), 2.28-2.25 (m, 1H), 2.15 (d, J = 0.9 Hz, 3H), 2.15-2.10 (m, 1H), 2.07-2.04 (m, 1H), 1.65-1.59 (m, 1H), 1.51-1.44 (m, 3H). 13 C NMR (300 MHz, methanol-d4) δ 163.7, 145.8, 142.6, 140.3, 137.0, 134.9, 133.8, 131.6, 131.3, 130.8, 129.6, 128.8, 124.3, 122.2, 121.1, 118.7, 113.9, 113.2, 58.7, 56.5, 49.6, 33.6, 30.1, 29.0, 22.6, 16.8. MS m / z (ESI): 428.2 [M+1].

[0153] Single configuration compound 9-2: 1 H NMR (600 MHz, methanol-d4) δ 8.04 (s, 1H), 7.61-7.58 (m, 2H), 7.57 (br, 1H), 7.56-7.53 (m, 2H), 7.47-7.45 (m, 1H), 7.41 (s, 1H), 7.29-7.27 (m, 1H), 6.60 (d, J = 9.2 Hz, 1H), 4.35 (d, J = 13.3 Hz, 1H), 4.29 (d, J = 13.3 Hz, 1H), 4.01-3.98 (m, 1H), 3.68-3.65 (m, 1H), 2.33-2.30 (m, 1H), 2.16-2.12 (m, 1H), 2.15 (d, J = 0.9 Hz, 3H), 2.00-1.97 (m, 1H), 1.88-1.74 (m, 5H). 13C NMR (300 MHz, methanol-d4) δ 163.7, 145.8, 142.6, 142.7, 139.5, 137.0, 134.5, 133.6, 131.6, 131.5, 129.8, 128.9, 124.5, 121.9, 121.1, 118.8, 113.0, 110.5, 55.2, 54.0, 49.6, 31.5, 28.7, 28.5, 19.6, 16.8. MS m / z (ESI): 428.2 [M+1].

[0154] Single configuration compounds 10-1 and 10-2 were prepared from 4-fluoropyridin-2-one 1c (Bide Pharm, CAS Number: 96530-75-5).

[0155] Single configuration compound 10-1: 1 H NMR (600 MHz, methanol-d4) δ 7.97 (s, 1H), 7.72 (t, J = 7.3 Hz, 1H), 7.61 (t, J = 7.8 Hz, 1H), 7.58 (dt, J = 7.8, 1.4 Hz, 1H), 7.54-7.52 (m, 2H), 7.44 (dt, J = 7.3, 1.4 Hz, 1H), 7.19 (d, J = 8.7 Hz, 1H), 7.47 (m, 1H), 7.33 (dd, J = 10.6, 2.8 Hz, 1H), 4.33 (d, J = 13.3 Hz, 1H), 4.27 (d, J = 13.3 Hz, 1H), 3.58-3.52 (m, 1H), 3.35-3.29 (m, 1H), 2.54-2.50 (m, 1H), 2.28-2.23 (m, 1H), 2.14-2.10 (m, 1H), 2.05-2.01 (m, 1H), 1.53-1.43 (m, 3H), 1.43-1.35 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 172.8 (d, J = 267.3 Hz), 165.7 (d, J = 20.2 Hz), 142.2 (d, J = 14.4 Hz), 141.9, 140.1, 138.8, 134.8, 133.9, 131.7, 131.6, 129.7, 128.9, 124.4, 122.1, 113.1, 104.0 (d, J = 17.3 Hz), 100.9 (d, J = 28.9 Hz), 109.0, 58.3.56.6, 49.6, 33.8, 30.2, 29.1, 22.7. MS m / z (ESI): 432.2 [M+1].

[0156] Single configuration compound 10-2:1 H NMR (600 MHz, methanol-d4) δ 8.02 (s, 1H), 7.71 (t, J = 7.8 Hz, 1H), 7.60-7.54 (m, 4H), 7.47-7.45 (m, 1H), 7.24 (d, J = 8.7 Hz, 1H), 6.47-6.44 (m, 1H), 7.33 (dd, J = 7.8, 2.8 Hz, 1H), 4.35 (d, J = 13.3 Hz, 1H), 4.28 (d, J = 13.3 Hz, 1H), 4.01-3.97 (m, 1H), 3.67-3.63 (m, 1H), 2.35-2.30 (m, 1H), 2.16-2.11 (m, 1H), 1.98-1.93 (m, 1H), 1.88-1.79 (m, 4H), 1.77-1.72 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 172.8 (d, J = 268.8 Hz), 165.8 (d, J = 18.8 Hz), 142.2 (d, J = 13.0 Hz), 141.8, 141.6, 139.2, 134.4, 133.7, 131.7, 131.7, 129.8, 129.0, 124.6, 121.8, 112.9, 109.4, 104.0 (d, J = 17.3 Hz), 101.0 (d, J = 27.4 Hz), 54.4, 54.2, 49.6, 31.6, 29.0, 28.6, 19.7. MS m / z (ESI): 432.2 [M+1].

[0157] Single configuration compounds 11-1 and 11-2 were prepared from 4- ethylpyridin-2(lH)-one lc (Bide Pharm, CAS Number: 37529-91-2).

[0158] Single configuration compound 11-1: 1H NMR (600 MHz, methanol-d4) δ 7.98 (s, 1H), 7.59 (t, J = 7.8 Hz, 1H), 7.56-7.55 (m, 2H), 7.52-7.50 (m, 2H), 7.43-7.42 (m, 1H), 7.22 (d, J = 8.7 Hz, 1H), 6.48 (d, J = 1.8 Hz, 1H), 6.44 (dd, J = 6.9, 1.8 Hz, 1H), 4.32 (d, J = 13.3 Hz, 1H), 4.26 (d, J = 13.3 Hz, 1H), 3.59-3.55 (m, 1H), 3.34-3.30 (m, 1H), 2.61 (q, J = 7.3 Hz, 2H), 2.52-2.49 (m, 1H), 2.27-2.24 (m, 1H), 2.13-2.10 (m, 1H), 2.05-2.02 (m, 1H), 1.54-1.39 (m, 4H), 1.26 (t, J = 7.8 Hz, 3H). 13 C NMR (300 MHz, methanol-d4) δ 164.7, 161.3, 142.4, 139.3, 139.2, 138.8, 134.4, 133.8, 131.6, 131.3, 129.6, 128.8, 124.6, 121.9, 118.3, 112.8, 110.4, 109.8, 56.8, 56.8, 49.1, 34.6, 31.0, 29.4, 29.3, 22.9, 13.8. MS m / z (ESI): 442.3 [M+1].

[0159] Single configuration compound 11-2: 1 H NMR (600 MHz, methanol-d4) δ 7.87 (s, 1H), 7.55 (t, J = 7.8 Hz), 7.52-7.50 (m, 2H), 7.46 (d, J = 7.3 Hz), 7.43 (dt, J = 7.8, 1.8 Hz, 1H), 7.42 (dd, J = 8.7, 0.9 Hz), 7.06 (d, J = 8.7 Hz, 1H), 6.47 (d, J = 1.87 Hz, 1H), 6.40 (dd, J = 7.3, 1.8 Hz, 1H), 4.30 (d, J = 13.3 Hz, 1H), 4.25 (d, J = 13.3 Hz, 1H), 3.95-3.93 (m, 1H), 3.58-3.55 (m, 1H), 2.60 (q, J = 7.8 Hz, 2H), 2.43-2.40 (m, 1H), 2.19-2.16 (m, 1H), 1.88-1.75 (m, 5H), 1.65-1.59 (m, 1H), 1.25 (t, J = 7.8 Hz, 3H). 13C NMR (300 MHz, methanol-d4) δ 164.7, 161.4, 142.4, 140.3, 139.2, 138.7, 134.4, 133.6, 131.6, 131.4, 129.8, 128.9, 124.6, 121.8, 118.3, 112.9, 110.4, 104.3, 54.5, 54.1, 49.6, 31.7, 29.4, 28.9, 28.7, 19.7, 13.8. MS m / z (ESI): 442.3 [M+1].

[0160] Single configuration compounds 13-1 and 13-2 were prepared from 4-trifluoromethylpyridin-2(lH)-one lc (Bide Pharm, CAS Number: 50650-59-4).

[0161] Single configuration compound 13-1: 1 H NMR (600 MHz, methanol-d4) δ 7.90 (s, 1H), 7.81 (d, J = 7.3 Hz, 1H), 7.64 (t, J = 7.8 Hz, 1H), 7.61-7.59 (m, 1H), 7.58 (t, J = 1.8 Hz, 1H), 7.51-7.49 (m, 1H), 7.46 (d, J = 8.7 Hz, 1H), 7.10 (d, J = 8.7 Hz, 1H), 6.94 (s, 1H), 6.64 (dd, J = 6.9, 1.8 Hz, 1H), 4.34 (d, J = 13.3 Hz), 4.29 (d, J = 13.3 Hz), 3.53-3.49 (m, 1H), 3.36-3.31 (m, 1H), 2.57-2.54 (m, 1H), 2.28-2.25 (m, 1H), 2.14-2.12 (m, 1H), 2.04-2.01 (m, 1H), 1.56-1.49 (m, 1H), 1.47-1.39 (m, 2H), 1.37-1.29 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 163.3, 143.5 (q, J = 30.3 Hz), 142.0, 141.8, 138.3, 138.0, 134.1, 133.9, 131.8, 131.8, 129.4, 128.7, 124.8, 123.4 (q, J = 283.2 Hz), 121.5, 119.4 (d, J = 4.3 Hz), 112.4, 106.3, 103.2, 57.1, 54.9, 48.9, 35.5, 31.9, 29.5, 23.1. MS m / z (ESI): 482.2 [M+1].

[0162] Single configuration compound 13-2: 1 H NMR (600 MHz, methanol-d4) δ 7.86 (s, 1H), 7.73 (d, J = 7.3 Hz), 7.55 (t, J = 7.8 Hz), 7.53-7.51 (m, 2H), 7.47 (dt, J = 7.3, 1.8 Hz, 1H), 7.33 (d, J = 8.7 Hz, 1H), 6.99 (d, J = 8.7 Hz, 1H), 6.91 (s, 1H), 6.57 (dd, J = 6.9, 2.3 Hz, 1H), 4.29 (d, J = 13.8 Hz, 1H), 4.24 (d, J = 13.8 Hz), 3.92-3.89 (m, 1H), 3.54-3.49 (m, 1H), 2.45-2.42 (m, 1H), 2.19-2.16 (m, 1H), 1.88-1.82 (m, 2H), 1.81-1.76 (m, 1H), 1.74-1.68 (m, 2H), 1.60-1.53 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 163.3, 143.7 (q, J = 33.2 Hz), 142.1, 142.0, 141.8, 141.9, 137.4, 134.0, 134.0, 131.9, 131.7, 129.4, 128.7, 124.4, 123.4 (q, J = 245.6 Hz), 121.1, 119.4 (d, J = 2.9 Hz), 112.3, 103.2, 101.6, 54.5, 50.2, 49.6, 32.9, 30.2, 29.6, 20.1. MS m / z (ESI): 482.2 [M+1].

[0163] Single configuration compounds 14-1 and 14-2 were prepared from 2-hydroxy-4-phenylpyridine 1c (Biotrend, CAS Number: 19006-81-6).

[0164] 19006-81-6).

[0165] Single configuration compound 14-1: 1H NMR (600 MHz, methanol-d4) δ 7.89 (s, 1H), 7.74 (dd, J = 7.3, 1.8 Hz, 2H), 7.68 (d, J = 7.3 Hz, 1H), 7.64 (t, J = 7.79, 1H), 7.60-7.58 (m, 2H), 7.52-7.49 (m, 4H), 7.45 (d, J = 8.7 Hz, 1H), 7.10 (d, J = 8.7 Hz, 1H), 6.89 (d, J = 1.8 Hz, 1H), 6.85 (dd, J = 7.3, 2.3 Hz, 1H), 4.35 (d, J = 12.8 Hz, 1H), 4.30 (d, J = 12.8 Hz, 1H), 3.54-3.49 (m, 1H), 3.38-3.33 (m, 1H), 2.26-2.56 (m, 1H), 2.28-2.26 (m, 1H), 2.14-2.12 m, 1H), 1.57-1.51 (m, 1H), 1.49-1.39 (m, 2H), 1.37-1.30 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 164.6, 155.4, 142.3, 139.6, 138.3, 138.3, 138.0, 134.0, 133.8, 131.7, 131.4, 131.3, 130.3, 129.6, 128.8, 128.0, 124.8, 121.5, 117.3, 112.4, 108.3, 106.2, 57.1, 54.9, 49.6, 35.5, 31.9, 29.5, 23.1. MS m / z (ESI): 490.3 [M+1].

[0166] Single configuration compound 14-2: 1H NMR (600 MHz, methanol-d4) δ 7.83 (s, 1H), 7.73-7.71 (m, 2H), 7.61 (d, J = 6.9 Hz, 1H), 7.58-7.56 (m, 2H), 7.53-7.49 (m, 5H), 7.36 (dd, J = 8.7, 0.9 Hz, 1H), 7.00 (d, J = 8.7 Hz, 1H), 6.88 (d, J = 1.8 Hz, 1H), 6.79 (d, J = 7.3, 1.8 Hz, 1H), 4.31 (d, J = 13.3 Hz, 1H), 4.27 (d, J = 13.3 Hz, 1H), 3.94-3.92 (m, 1H), 3.56-3.52 (m, 1H), 2.47-2.45 (m, 1H), 2.21-2.19 (m, 1H), 1.90-1.85 (m, 2H), 1.84-1.78 (m, 1H), 1.77-1.72 (m, 2H), 1.62-1.56 (m, 1H). 13 C NMR (300 MHz, methanol-d4) δ 164.6, 155.3, 142.3, 142.1, 139.6, 138.0, 137.2, 133.8, 133.4, 131.6, 131.5, 131.2, 130.3, 129.6, 128.8, 127.9, 125.0, 121.1, 117.2, 112.2, 108.2, 101.7, 54.5, 50.2, 49.6, 32.9, 30.2, 29.6, 20.2. MS m / z (ESI): 490.3 [M+1].

[0167] Test Example 1

[0168] The following method is used to determine the inhibitory effect of the compounds of the present disclosure on ROCK2 kinase activity. The experimental method is briefly described as follows: ROCK2 kinase inhibition activity is determined by HTRF KinEASE kit (PerkinElmer, Cat No: 62ST2PEB) based on time-resolved fluorescence technology. KD025 (MCE, CAS No: 911417-87-3) is used as a positive control, and three replicate wells are set up. In a white 384-well plate, 4 μL of the test compound or buffer, 2 μL of the reaction substrate S2, 2 μL of ROCK2 kinase, and 2 μL of ATP are added to each well in turn. Incubate in a 37°C incubator for 30 min. Add 5 μL of streptokinase-labeled XL-665 and 5 μL of EuK-labeled anti-phosphorylated protein kinase antibody in turn, and react at room temperature for 60 min. The fluorescence signal is detected using a Tecan multifunctional microplate reader. The fluorescence intensity value of the sample well at λ = 665 nm is F1, and the fluorescence intensity value at λ = 620 nm is F2. The signal ratio calculation formula is: signal ratio = F1 / F2 x 10 4 . The inhibition rate of the test compound on protein kinase is calculated by the formula: inhibition rate (%) = [1-(test compound signal ratio-minimum signal ratio) / (maximum signal ratio-minimum signal ratio)] x 100. The maximum signal ratio is the solvent control, and the minimum signal ratio is the signal ratio of the well without streptokinase-labeled XL-665. The IC 50 values of the inhibitory activity of the compounds are calculated using GraphPad Prism 8 software, as shown in Table 1.

[0169] Table 1 Inhibitory activity of the compounds of the present disclosure on ROCK2 kinase activity

[0170] Compound IC 50 (nM) Compound IC 50 (nM) 1-1 83.73±6.64 9-1 137.95±8.70 1-2 66.15±2.51 9-2 35.40±2.21 2-1 77.51±12.49 10-1 88.11±14.81 2-2 53.97±3.31 10-2 36.64±9.79 3-1 433.70±12.59 11-1 6.06±1.16 3-2 560.85±3.75 11-2 13.09±3.97 4-1 11.88±2.75 12-1 27.04±0.33 4-2 25.98±3.44 12-2 3.68±0.76 5-1 73.72±9.20 13-1 206.65±12.23 5-2 42.41±5.59 13-2 173.15±21.00 6-1 77.10±12.63 14-1 1.68±0.69 6-2 65.49±3.17 14-2 2.97±0.69 7-1 130.35±3.61 15-1 1.55±0.55 7-2 101.50±9.62 15-2 1.30±0.88 8-1 152.70±16.12 KD025 119.50±8.06 8-2 18.68±5.03

[0171] Conclusion: The compounds of the present disclosure have good inhibitory activity on ROCK2 kinase.

[0172] Test Example 2

[0173] The inhibitory effect of the compounds of the present disclosure on ROCK1 kinase activity is determined, and the experimental method is the same as that of Test Example 1, but the incubation time is changed to 20 min. The test results are shown in Table 2.

[0174] Table 2 Inhibitory activity of the compounds of the present disclosure on ROCK1 kinase activity

[0175] Compound IC 50 (nM) Compound IC 50 (nM) 4-1 812.25±21.43 10-1 272.75±9.12 4-2 170.85±21.99 10-2 206.65±5.16 5-1 643.80±9.76 11-1 312.60±24.18 5-2 161.65±16.33 11-2 209.40±8.91 6-1 272.10±0.42 12-1 215.70±14.42 6-2 285.20±16.69 12-2 71.20±11.26 7-1 464.00±3.39 13-1 966.55±18.17 7-2 237.80±1.98 13-2 279.50±6.79 8-1 811.05±3.61 14-1 99.38±0.17 8-2 277.15±4.45 14-2 96.13±2.75 9-1 102.43±12.97 15-1 335.05±3.61 9-2 152.40±15.41 15-2 209.90±3.39 KD025 3259.50±98.29

[0176] Conclusion: The compounds of the present disclosure have good inhibitory activity on ROCK1 kinase.

Claims

1. An indazole derivative or its pharmaceutical salt, characterized in that, The indazole derivative is any one of the following compounds: 。 2. The use of the indazole derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, is any one of the following: (1) use in the manufacture of a ROCK kinase inhibitor; (2) use in the manufacture of a medicament for treating and / or preventing a disease or disorder associated with ROCK kinase, the disease or disorder being selected from the group consisting of inflammatory diseases, autoimmune diseases, fibrotic diseases, transplant rejection, diseases associated with overproduction of IL-17, IL-21 and / or IL-23, ocular diseases, cardiovascular and cerebrovascular diseases, tumor metastasis, central nervous system diseases, diabetic nephropathy and dermatological diseases.

3. Use according to claim 2, characterized in that, The disease or disorder is selected from the group consisting of rheumatoid arthritis, osteoarthritis, juvenile idiopathic arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, allergic airway disease, chronic obstructive pulmonary disease, asthma, bronchitis, inflammatory bowel disease, systemic lupus erythematosus, cutaneous lupus erythematosus, lupus nephritis, dermatomyositis, autoimmune liver disease, Sjogren's syndrome, multiple sclerosis, dry eye disease, type I diabetes and complications associated therewith, atopic eczema, thyroiditis, contact dermatitis, Sjogren's syndrome, and amyotrophic lateral sclerosis.

4. A medicament, characterized by comprising: The active ingredient is the indazole derivative of claim 1 or a pharmaceutically acceptable salt thereof.

5. A process for preparing the indazole derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized by, The reaction formula is: ， wherein the compound represented by general formula (IA) or a salt thereof is subjected to reductive amination to obtain the compound represented by general formula (I) or a pharmaceutically acceptable salt thereof; G, R 1 , R 2 , R 3 , a, q, s, ring A all correspond to the indazole derivative or pharmaceutically acceptable salt thereof of claim 1.

6. The preparation method according to claim 5, characterized in that, The preparation method of general formula (IA) is: ， wherein the compound represented by general formula (IA-1) and general formula (IA-2) or a salt thereof is subjected to coupling reaction to obtain the compound represented by general formula (IA) or a salt thereof.

7. The preparation method according to claim 5, characterized in that, The preparation method of general formula (IV-1) is: ， wherein the compound represented by general formula (V-1) or a salt thereof is subjected to deprotection to obtain the compound represented by general formula (IV-1) or a salt thereof.

8. The preparation method according to claim 7, characterized in that, The preparation method of general formula (V-1) is: ， wherein: The compound represented by general formula (IV-2) and general formula (IV-4) or a salt thereof is subjected to reductive amination to obtain the compound represented by general formula (V-1) or a salt thereof.

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