Indazole derivative or pharmaceutical salt and application thereof

Abstract

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

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an indazole derivative or a pharmaceutically acceptable salt thereof and application thereof. Background Art

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

[0003] ROCK kinases are involved in the occurrence and development of many diseases, such as eye diseases, cardiovascular and cerebrovascular diseases, tumor metastasis, central nervous system diseases, diabetic nephropathy, etc., and are important targets for the treatment of many diseases. Rho GTPase-mediated signaling pathways play an important role in balancing and coordinating T cell-mediated immune processes, including the development, activation, and differentiation of T cells. Recent studies have shown that overactivation of ROCK kinases also plays an indispensable role in inflammatory and autoimmune diseases, such as psoriasis, inflammatory bowel disease, rheumatoid arthritis, systemic lupus erythematosus, etc. Although dysregulated ROCK kinases mainly affect 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 have shown that ROCK2 regulates the production of IL-21 and IL-17 in mice by phosphorylating interferon regulatory factor 4 (IRF4), playing an important role in the development of autoimmunity. In human peripheral blood CD4 +In the Th17 cells produced by T cell polarization, ROCK2 binds to phosphorylated STAT3 to regulate Th17 cell-specific gene transcription. A clinical study showed that oral 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). Two other studies showed that targeting ROCK2 inhibits STAT3 phosphorylation and increases STAT5 phosphorylation, thereby downregulating overactivated Th17 cells and enhancing regulatory T cell (Treg) function, thereby rebuilding immune balance (Blood. 2016, 127, 2144-2154). Studies by Tenggesdal et al. showed that KD025 downregulates the secretion of IL-17 independently of IL-1 and IL-6 (Eur. J. Immunol. 2018, 48, 1679-1686). In non-blood cells, ROCK protein is also involved in the TGF-β signaling pathway that controls the balance of Th17 cells and Treg cells (Nat. Commun. 2020, 11, 2608). Based on the role of ROCK2 in inhibiting the overactivation 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. It is mainly used to treat glaucoma, cerebral vasospasm and cerebral ischemia. It also has good effects in treating amyotrophic lateral sclerosis (ALS), inducing tumor cell apoptosis, inhibiting tumor metastasis, and alleviating symptoms of neurological diseases (such as Parkinson's disease). Ripasudil was approved for marketing in Japan in 2014, mainly for the treatment of glaucoma and ocular hypertension. Its IC50 for inhibiting ROCK2 and ROCK1 is 19 and 51 nM, respectively. Netarsudil was approved by the FDA in 2017 for the treatment of glaucoma or ocular hypertension, and its Ki value for inhibiting ROCK2 is 2 nM. 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. The compound was approved by the FDA for the treatment of graft-versus-host disease in 2021. In addition, there are 7 compounds in the clinical research stage, mainly used to treat diseases such as glaucoma, spinal cord injury and pulmonary fibrosis. Summary of the invention

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

[0006] The present invention first provides an indazole derivative or a pharmaceutically acceptable salt thereof, wherein the structural formula of the indazole derivative is shown in general formula (I):

[0007]

[0008] in:

[0009] G is a CH or N atom;

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

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

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

[0013] Each R 3 are the same or different and are each independently selected from hydrogen, halogen, an alkyl group having 1 to 6 carbon atoms, a haloalkyl group having 1 to 6 carbon atoms, a haloalkoxy group having 1 to 6 carbon atoms, a cyano group, a hydroxyl group, a nitro group, a carboxyl group, and a 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, "selected from" means selecting one from a plurality of selectable groups.

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

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

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

[0021] Preferably, the structural formula of the indazole derivative is as shown in general formula (I-1), (I-2), (I-3) or (I-4),

[0022]

[0023] Preferably, ring A is selected from 5- to 7-membered cycloalkyl, heterocyclic and benzene rings.

[0024] More preferably, ring A is selected from cyclopentane, cyclohexane, cycloheptane, benzene ring and pyridine ring, and the structural formula of the indazole derivative is as shown in general formula (II-1), (II-2) or (II-3),

[0025]

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

[0027] Further preferably, the structural formula of the indazole derivative is as shown in 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 the chiral cis and trans forms, respectively.

[0033] The present invention further provides the use of the indazole derivatives or pharmaceutically acceptable salts thereof in the preparation of ROCK kinase inhibitors.

[0034] The present invention further provides the use of the indazole derivatives or pharmaceutically acceptable salts thereof in the preparation of a medicament for treating and / or preventing a disease or condition, wherein the disease or condition is selected from inflammatory diseases, autoimmune diseases, fibrotic diseases, transplant rejection, diseases associated with 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 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, Sjögren's syndrome, multiple sclerosis, dry eye disease, type I diabetes and complications associated therewith, atopic eczema, thyroiditis, contact dermatitis, Sjögren's syndrome and amyotrophic lateral sclerosis.

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

[0037] The present invention also provides a method for preparing the indazole derivative or a pharmaceutically acceptable salt thereof, and the reaction formula is:

[0038]

[0039] The compounds represented by the general formula (IA) and the general formula (IV-1) or their salts undergo a reductive amination reaction to obtain the compound represented by the general formula (I) or its pharmaceutically acceptable salt.

[0040] The preparation method also includes the preparation method of the general formula (IA):

[0041]

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

[0043] The preparation method also includes the preparation method of the general formula (IV-1):

[0044]

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

[0046] The preparation method also includes the preparation method of the general formula (IV-1):

[0047]

[0048] Wherein, the compounds represented by the general formula (IV-2) and the general formula (IV-4) or their salts undergo a reductive amination reaction to obtain the compound represented by the general formula (V-1) or their salts. Alternatively, the compounds represented by the general formula (IV-3) and the general formula (IV-4) or their salts undergo a coupling reaction to obtain the compound represented by the general formula (V-1) or their salts.

[0049] The present invention has discovered a new class of indazole derivatives, which have an inhibitory function on ROCK kinase and can be used as a ROCK kinase inhibitor. Moreover, the structural type of the indazole derivatives is relatively novel and can be used as a potential therapeutic drug for a variety of 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. DETAILED DESCRIPTION

[0050] The following examples are used to further describe the present disclosure, but these examples do not limit the scope of the present disclosure. The indazole derivative of the present invention is any one of the following compounds:

[0051]

[0052]

[0053] Example 1

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

[0055]

[0056] The first step is to synthesize 1b:

[0057] Under ice bath, sodium hydride (567 mg, 23.6 mmol) was added to anhydrous tetrahydrofuran (80 mL), 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 and quenching with saturated ammonium chloride, the solvent was removed by concentration under reduced pressure, and the residual liquid was extracted with dichloromethane, which was repeated three times and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. 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] 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. The mixture was stirred at room temperature for 2 hours under a hydrogen atmosphere, filtered through celite, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography with dichloromethane / methanol (20:1, v / v, 500 mL) to give the title compound 1c in a yield of 90-95%.

[0060] The third step is to synthesize tert-butyl ((1S)-3-((1H-indazol-5-yl)amino)cyclohexyl)carbamate 1f:

[0061] (S)-(3-carbonylcyclohexyl)carbamic acid tert-butyl ester 1d (Bidder Pharmaceuticals, CAS No.: 1803033-61-5) (1.00 g, 4.69 mmol), 5-aminoindazole 1e (Bidder Pharmaceuticals, CAS No.: 19335-11-6) (750 mg, 5.63 mmol), sodium cyanoborohydride (500 mg, 7.96 mmol) were added to anhydrous methanol (50 mL), and the pH was adjusted to 6.0-7.0 with acetic acid. The mixture was reacted at room temperature for 12 hours. Water (2 mL) was added, and the solvent was removed by concentration under reduced pressure. The residue was purified by silica gel column chromatography with dichloromethane / methanol (40:1, v / v, 500 mL) containing 1% triethylamine by volume to obtain the title compound 1f (1.42 g), with a yield of 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] Compound 1f (1.42 g, 4.30 mmol) and trifluoroacetic acid (10 ml) were dissolved in dichloromethane (10 ml) and reacted at room temperature for 1 hour. The reaction solution was concentrated under 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 to obtain the title compound 1g (815.7 mg) with a yield of 82%.

[0065] After mass spectrometry analysis of 1 g of the obtained product, m / z (ESI): 231.2 [M+1].

[0066] Step 5: Synthesis of 1i:

[0067] Compound 1c (2.36 mmol) with different substituents, 3-formylphenylboronic acid 1h (Bidec Pharmaceuticals, CAS No.: 87199-16-4) (708 mg, 4.72 mmol), copper acetate (944 mg, 4.72 mmol), pyridine (4.5 mL) were added to 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 to obtain the corresponding compound 1i with a yield of 54-91%.

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

[0069] 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. The mixture was reacted 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, 21.2×250mm, 5μm; elution system: methanol / water (volume ratio) = 10%-100%, containing 0.5‰ trifluoroacetic acid by volume, flow rate: 10ml / min, elution time: 40min) to obtain the corresponding compounds II-1 and II-2 with yields of 30-32% and 61-63%, respectively.

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

[0071] Single configuration compound 12-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.92(s, 1H), 7.65(d, J=5.0Hz, 1H), 7.60(t, J=7.8Hz, 1H), 7.57-7.54(m, 2H), 7.49-7.45(m, 2H), 7.12 (d, J=8.7Hz, 1H), 6.53 (d, J=6.9Hz, 1H), 4.32 (d, J=13.3Hz, 1H), 4.27 (d, J=13.3Hz, 1 H), 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). 13 CNMR (300MHz, methanol-d 4) δ162.4, 149.8 (d, J = 232.6Hz), 146.8 (d, J = 20.2Hz), 141.9, 138.5, 137.5, 134.0, 133.9, 131.6, 131.4, 129.6, 128.8, 124.7, 124.2 (d, J=39.0Hz), 121.6, 121.0 (d, J=2.9Hz), 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-d 4 )δ7.82 (s, 1H), 7.60 (d, J=4.6Hz, 1H), 7.54 ((t, J=7.8Hz, 1H), 7.51-7.49 (m, 2H), 7.46-7. 44 (m, 1H), 7.35 (d, J = 9.6Hz, 1H), 6.98 (d, J = 8.7Hz), 6.51 (dd, J = 7.3, 0.9Hz), 4.29 (d, J = 13 .3Hz, 1H), 4.24 (d, J=13.3Hz, 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 (300MHz, methanol-d 4 ) δ162.4, 149.8 (d, J = 234.1Hz), 146.8 (d, J = 20.2Hz), 142.2, 141.9, 137.1, 133.8, 133.3, 131.6, 131.6, 129.6, 128.7, 125.0, 124.2 (d, J=39.0Hz), 121.1, 121.0 (d, J=4.3Hz), 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 1a (Bidec Pharmaceuticals, CAS No.: 81167-60-4).

[0074] Single configuration compound 15-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.91 (s, 1H), 7.61 (t, J=7.8Hz, 1H), 7.57-7.55 (m, 1H), 7.54-7.52 (m, 2H), 7.48 (d, J=9.2Hz, 1H), 7.45-7 .43 (m, 1H), 7.12 (d, J = 8.7Hz, 1H), 6.63 (dd, J = 7.3, 2.3Hz, 1H), 6.58 (d, J = 1.8Hz, 1H), 4.33 (d, J = 13.3Hz, 1 H), 4.27 (d, J=13.3Hz, 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 (300MHz, methanol-d 4 )δ167.7, 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. MSm / z(ESI): 470.3[M+1].

[0075] Single configuration compound 15-2: 1 H NMR (600 MHz, methanol-d 4)δ7.85 (s, 1H), 7.55 (t, J=7.8Hz, 1H), 7.52-7.49 (m, 2H), 7.47 (d, J=8.2Hz, 1H), 7.45 -7.43 (m, 1H), 7.39 (d, J = 8.7Hz, 1H), 7.03 (d, J = 9.2Hz, 1H), 6.58-6.56 (m, 2H), 4.30 ( d, J=13.3Hz, 1H), 4.26 (d, J=13.3Hz, 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). 13 C NMR (300MHz, methanol-d 4 )δ167.7, 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. MSm / z(ESI): 470.3[M+1].

[0076] Example 2

[0077] The compounds in Example 2 are all obtained by the following reaction formula:

[0078]

[0079] The first step is to synthesize 1k:

[0080] 1j (5.0 mmol) with different substituents, 1h (10.0 mmol) with para- or meta-substituted aldehyde groups, copper acetate monohydrate (998 mg, 5.0 mmol), and tetramethylethylenediamine (1.5 mL) were added to 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 to obtain the title compound 1k (297.7 mg) with a yield of 63%.

[0081] The second step is to synthesize 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 to anhydrous methanol (20 mL), and the pH was adjusted to 6.0-7.0 with acetic acid. The mixture was reacted 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, 21.2×250mm, 5μm; elution system: methanol / water (volume ratio) = 10%-100%, containing 0.5‰ trifluoroacetic acid by volume, flow rate: 10ml / min, elution time: 40min) 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 (Bidec Pharmaceuticals, CAS No.: 2022-85-7).

[0084] Single configuration compound 1-1: 1 H NMR (600 MHz, methanol-d 4 )δ8.06(s, 1H), 7.90(d, J=6.0Hz, 1H), 7.65-7.61(m, 2H), 7.56-7.52(m, 2H), 7.48(d , J=7.8Hz, 1H), 7.40 (dq, J=7.8, 0.9Hz, 1H), 7.30 (dd, J=9.2, 2.3Hz, 1H), 4.31 (d, J=1 2.8Hz, 1H), 4.24d, J=12.8Hz, 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 (300MHz, methanol-d 4) δ160.1 (d, J=15.9Hz), 156.4, 142.0, 139.8, 138.8 (d, J=244.2Hz), 134.7, 133.6, 131.6, 131.3 (d, J=33. 2Hz), 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-d 4 )δ8.06 (s, 1H), 7.95 (d, J = 6.0Hz, 1H), 7.61 (d, J = 8.7Hz, 1H), 7.58-7.51 (m, 3H), 7.47-7.45 (m, 1H), 7.29 (dd, J=8.7, 1.8Hz, 1H), 4.34 (d, J=13.3H z, 1H), 4.27 (d, J=13.3Hz, 1H), 4.02-3.98 (m, 1H), 3.69-3.64 (m, 1H), 2.3 2-2.27(m, 1H), 2.15-2.10(m, 1H), 2.03-1.96(m, 1H), 1.90-1.74(m, 5H). 13 C NMR (300MHz, methanol-d 4 )δ159.9 (d, J=15.9Hz), 156.1, 141.0, 138.8, 138.7 (d, J=244.2Hz), 137.9, 134.2, 133.5, 131.5, 131.4 (d , J=31.8Hz), 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] The single-configuration compounds 16-1 and 16-2 were prepared from cytosine 1j (Bidec Pharmaceuticals, CAS No.: 71-30-7).

[0087] Single configuration compound 16-1: 1 H NMR (600 MHz, methanol-d 4)δ8.04 (s, 1H), 7.90 (d, J = 7.8Hz, 1H), 7.63 (d, J = 8.2Hz, 2H), 7.60 (d, J = 9.2Hz , 1H), 7.49 (d, J = 8.2Hz, 2H), 7.30-7.27 (m, 1H), 6.18 (d, J = 7.8Hz, 1H), 4.34 (d , J=13.3Hz, 1H), 4.29 (d, J=13.3Hz, 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-d 4 )δ162.3, 150.2, 149.4, 140.5, 139.4, 134.5, 134.2, 133.7, 132.3, 128.3, 124.5, 12 1.9, 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: 1 H NMR (600 MHz, methanol-d 4 )δ8.21 (s, 1H), 7.75 (d, J = 7.8Hz, 1H), 7.43 (d, J = 8.2Hz, 2H), 7.31 (d, J = 9.2Hz , 1H), 7.25 (d, J = 8.2Hz, 2H), 7.22-7.19 (m, 1H), 6.15 (d, J = 7.8Hz, 1H), 4.22 (d , J=13.3Hz, 1H), 4.15 (d, J=13.3Hz, 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-d 4)δ165.1, 152.4, 148.4, 142.4, 138.2, 135.4, 134.4, 133.6, 132.4, 127.2, 123.4, 12 2.2, 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+1].

[0089] The single-configuration compounds 17-1 and 17-2 were prepared from cytosine 1j (Bidec Pharmaceuticals, CAS No.: 71-30-7).

[0090] Single configuration compound 17-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.75(s, 1H), 7.49(d, J=6.9Hz, 2H), 7.33(d, J=8.7Hz, 1H), 7.11(brs, 1H ), 6.89 (dd, J=8.7, 2.3Hz, 1H), 6.51 (brs, 1H), 6.09 (d, J=7.8Hz, 1H), 4.45 (d, J=11.9Hz, 1H), 4.32 (brs, 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). 13 C NMR (150 MHz, methanol-d 4 )δ162.0, 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-d 4)δ7.78(s, 1H), 7.45(d, J=6.9Hz, 2H), 7.37(d, J=8.7Hz, 1H), 7.21(brs, 1H ), 6.65 (dd, J=8.7, 2.3Hz, 1H), 6.53 (brs, 1H), 6.19 (d, J=7.8Hz, 1H), 4.54 (d, J=11.9Hz, 1H), 4.37 (brs, 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-d 4 )δ162.4, 148.4, 148.1, 146.6, 142.1, 136.1, 132.3, 132.1, 131.0, 127.9, 124.5121. 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 (Bidec Pharmaceuticals, CAS No.: 71-30-7).

[0093] Single configuration compound 18-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.86 (s, 1H), 7.76 (d, J=7.3Hz, 1H), 7.55-7.50 (m, 3H), 7.46 (dt, J=6.9, 1.8H z, 1H), 7.39 (d, J=8.7Hz, 1H), 7.03 (d, J=8.7Hz, 1H), 6.09 (d, J=7.3Hz, 1H), 4.3 0 (d, J=13.3Hz, 1H), 4.25 (d, J=13.3Hz, 1H), 3.94-3.91 (m, 1H), 3.57-3.52 (m, 1 H), 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-d 4)δ164.6, 153.0, 148.8, 141.0, 139.4.137.5, 133.9, 133.5, 131.6, 131.5, 129.4, 1 28.5, 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-d 4 )δ7.83 (s, 1H), 7.74 (d, J = 7.3Hz, 1H), 7.65-7.60 (m, 3H), 7.48 (dt, J = 6.9, 1.8H z, 1H), 7.33 (d, J=8.7Hz, 1H), 7.05 (d, J=8.7Hz, 1H), 6.03 (d, J=7.3Hz, 1H), 4.3 2 (d, J=13.3Hz, 1H), 4.25 (d, J=13.3Hz, 1H), 3.84-3.71 (m, 1H), 3.64-3.60 (m, 1 H), 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-d 4 )δ164.5, 153.8, 148.5, 141.5, 139.2137.7137.9, 134.5, 132.6, 135.5, 129.8, 12 8.9, 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] The single-configuration compounds 19-1 and 19-2 were prepared from cytosine 1j (Bidec Pharmaceuticals, CAS No.: 71-30-7).

[0096] Single configuration compound 19-1: 1 H NMR (600 MHz, methanol-d 4)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 (brs, 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 (brs, 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 - d 4 )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: 1 H NMR (600 MHz, methanol - d 4 )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 (brs, 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 (brs, 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 - d 4 )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 + 1].

[0099] The single-configuration compound 20-1 was prepared from cytosine 1j (Biode Pharmaceuticals, CAS No.: 71-30-7).

[0100] Single configuration compound 20-1: 1 H NMR (600 MHz, DMSO-d 6 ) δ9.59 (s, 1H), 8.56 (d, J = 6.9Hz, 1H), 8.43 (d, J = 6.9Hz, 1H), 7.90 (d, J = 7.3Hz, 1H), 7.87 (d, J = 7.8Hz, 1H), 7.75 (t, J = 8.2Hz, 1H), 7.71 (d, J = 7. 8Hz, 1H), 7.47 (t, J=8.2Hz, 1H), 7.46-7.44 (m, 2H), 7.35 (dd, J=7.3, 1.8Hz, 1H), 7.08 (s, 1H), 6.40 (s, 1H), 6.14 (d, J=7.8Hz, 1H), 4.38 (s, 2H). 13 C NMR (150 MHz, DMSO-d 6 )δ165.5, 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] The single-configuration compound 21-1 was prepared from cytosine 1j (Bidec Pharmaceuticals, CAS No.: 71-30-7).

[0102] Single configuration compound 21-1: 1 H NMR (600 MHz, DMSO-d 6 )δ9.59 (s, 1H), 8.56 (d, J = 6.9Hz, 1H), 8.43 (d, J = 6.9Hz, 1H), 7.90 (d, J = 7.8Hz, 1H), 7.87 (d, J = 7.8Hz, 1H), 7.75 (t, J = 8.2Hz, 1H), 7 .71 (d, J=7.8Hz, 1H), 7.46-7.44 (m, 2H), 7.35 (dd, J=7.3, 1.8Hz, 1H), 7.08 (s, 1H), 6.40 (s, 1H), 6.14 (d, J=7.8Hz, 1H), 4.38 (s, 2H). 13 C NMR (150 MHz, DMSO-d6 )δ166.5, 153.0, 147.5, 142.8, 134.2, 135.1, 134.4, 133.5, 124.1, 127.1, 127.0, 1 25.1, 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] The single-configuration compound 22-1 was prepared from cytosine 1j (Bidec Pharmaceuticals, CAS No.: 71-30-7).

[0104] Single configuration compound 22-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.82 (s, 1H), 7.75 (d, J=7.3Hz, 1H), 7.66-7.55 (m, 2H), 7.49 (s, 1H), 7.34 (d, J=8.7Hz, 1H), 7.04 (d, J=8.7Hz, 1H), 6.03 (d, J=7.3Hz, 1H), 4.32 (d, J=1 3.3Hz, 1H), 4.25 (d, J=13.3Hz, 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-d 4 )δ164.4, 153.1, 148.5, 141.6, 139.7137.8137.2, 134.4, 132.5, 135.5, 129.8, 12 8.3, 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+1].

[0105] The single-configuration compound 23-1 was prepared from cytosine 1j (Biode Pharmaceuticals, CAS No.: 71-30-7).

[0106] Single configuration compound 23-1: 1 H NMR (600 MHz, methanol-d 4)δ7.83 (s, 1H), 7.75 (d, J=7.3Hz, 1H), 7.66-7.53 (m, 2H), 7.49 (s, 1H), 7.39 (d, J=8.7Hz, 1H), 7.09 (d, J=8.7Hz, 1H), 6.04 (d, J=7.3Hz, 1H), 4.37 (d, J=1 3.3Hz, 1H), 4.28 (d, J=13.3Hz, 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). 13 C NMR (150 MHz, methanol-d 4 )δ166.4, 154.1, 147.5, 142.6, 139.5 137.7137.3, 134.3, 132.5, 135.8, 129.8, 128.4, 125.7, 121.2, 112.1, 112.2, 96.0, 51.7, 50.8, 49.8, 32.7, 30.1, 29.8, 20.8. MS m / z(ESI): 448.2[M+1].

[0107] The single-configuration compound 24-1 was prepared from cytosine 1j (Biode Pharmaceuticals, CAS No.: 71-30-7).

[0108] Single configuration compound 24-1: 1 H NMR (600 MHz, methanol-d 4 )δ8.12(s, 1H), 7.78(d, J=7.8Hz, 1H), 7.72-7.64(m, 2H), 7.14(s, 1H), 6.91 (d, J=8.5Hz, 1H), 6.62 (d, J=8.5Hz, 1H), 6.14 (d, J=7.8Hz, 1H), 4.39 (d, J=1 2.0Hz, 1H), 4.38 (d, J=12.0Hz, 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-d 4)δ167.4, 155.1, 147.9, 144.3, 138.4137.3137.1, 134.4, 132.5, 135.9, 129.1, 12 8.6, 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] The single-configuration compound 25-1 was prepared from cytosine 1j (Biode Pharmaceuticals, CAS No.: 71-30-7).

[0110] Single configuration compound 25-1: 1 H NMR (600 MHz, methanol-d 4 )δ8.20(s, 1H), 7.52(d, J=7.8Hz, 1H), 7.70-7.63(m, 2H), 7.10(s, 1H), 6.81(d , J=8.7Hz, 1H), 6.32 (d, J=8.7Hz, 1H), 6.04 (d, J=7.3Hz, 1H), 4.38 (d, J=13.3Hz , 1H), 4.28 (d, J=13.3Hz, 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-d 4 )δ166.4, 154.1, 147.5, 142.6, 139.5137.7137.3, 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+1].

[0111] The single-configuration compound 26-1 was prepared from cytosine 1j (Biode Pharmaceuticals, CAS No.: 71-30-7).

[0112] Single configuration compound 26-1: 1 H NMR (600 MHz, methanol-d 4)δ8.32(s, 1H), 8.12(d, J=7.8Hz, 1H), 7.87-7.72(m, 2H), 7.52(s, 1H), 6.98 (d, J=8.7Hz, 1H), 6.45 (d, J=8.7Hz, 1H), 6.21 (d, J=7.3Hz, 1H), 4.39 (d, J=1 3.3Hz, 1H), 4.32 (d, J=13.3Hz, 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). 13 C NMR (150 MHz, methanol-d 4 )δ168.2, 155.2, 145.5, 144.6, 137.5135.7133.3, 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] The single-configuration compound 27-1 was prepared from cytosine 1j (Biode Pharmaceuticals, CAS No.: 71-30-7).

[0114] Single configuration compound 27-1: 1 H NMR (600 MHz, methanol-d 4 )δ8.15(s, 1H), 7.92(d, J=7.8Hz, 1H), 7.82-7.810(m, 2H), 7.75(s, 1H), 7.42(d , J=8.5Hz, 1H), 6.63 (d, J=8.5Hz, 1H), 6.54 (d, J=7.8Hz, 1H), 4.33 (d, J=12.0Hz , 1H), 4.30 (d, J=12.0Hz, 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-d 4)δ173.2, 169.2, 159.1, 148.9, 148.3, 141.4138.2137.2, 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] The single-configuration compound 28-1 was prepared from cytosine 1j (Bidec Pharmaceuticals, CAS No.: 71-30-7).

[0116] Single configuration compound 28-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.82 (d, J=7.5Hz, 1H), 7.57-7.52 (m, 3H), 7.50 (dt, J=6.9, 1.8Hz, 1H), 7. 35 (d, J=8.5Hz, 1H), 7.02 (d, J=8.5Hz, 1H), 6.59 (d, J=7.5Hz, 1H), 4.33 (d, J= 13.1Hz, 1H), 4.23 (d, J=13.1Hz, 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-d 4 )δ168.3, 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] The single-configuration compound 29-1 was prepared from cytosine 1j (Bidec Pharmaceuticals, CAS No.: 71-30-7).

[0118] Single configuration compound 29-1: 1 H NMR (600 MHz, methanol-d 4)δ7.72 (d, J=7.3Hz, 1H), 7.54-7.49 (m, 3H), 7.42 (dt, J=6.9, 1.8Hz, 1H), 7.35 ( d, J=8.7Hz, 1H), 7.04 (d, J=8.7Hz, 1H), 6.08 (d, J=7.3Hz, 1H), 4.35 (d, J=13.3H z, 1H), 4.29 (d, J=13.3Hz, 1H), 3.97-3.90 (m, 1H), 3.58-3.51 (m, 1H), 2.44-2.4 0(m, 1H), 2.25-2.14(m, 1H), 2.06(s, 3H), 1.91-1.84(m, 5H), 1.67-1.59(m, 1H). 13 C NMR (150 MHz, methanol-d 4 )δ167.6, 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] The single-configuration compound 30-1 was prepared from cytosine 1j (Biode Pharmaceuticals, CAS No.: 71-30-7).

[0120] Single configuration compound 30-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.86 (d, J=7.3Hz, 1H), 7.52-7.45 (m, 3H), 7.41 (dt, J=6.9, 1.8Hz, 1H), 7. 39 (d, J=8.7Hz, 1H), 7.23 (d, J=8.7Hz, 1H), 6.49 (d, J=7.3Hz, 1H), 4.32 (d, J= 12.0Hz, 1H), 4.25 (d, J=12.0Hz, 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-d 4)δ164.6, 153.0, 148.8, 141.0, 139.4.137.5, 133.9, 133.5, 131.6, 131.5, 129.4, 1 28.5, 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] The single-configuration compound 31-1 was prepared from cytosine 1j (Bidec Pharmaceuticals, CAS No.: 71-30-7).

[0122] Single configuration compound 31-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.96 (d, J=7.8Hz, 1H), 7.57-7.51 (m, 3H), 7.39 (dt, J=6.9, 1.8Hz, 1H), 7. 35 (d, J=8.7Hz, 1H), 7.23 (d, J=8.7Hz, 1H), 6.19 (d, J=7.3Hz, 1H), 4.30 (d, J= 13.3Hz, 1H), 4.25 (d, J=13.3Hz, 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-d 4 )δ169.6, 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 by the following reaction formula:

[0125]

[0126] The first step is to synthesize 1i:

[0127] Compound 1c (2.36 mmol) with different substituents, 3-formylphenylboronic acid 1h (Bidec Pharmaceuticals, CAS No.: 87199-16-4) (708 mg, 4.72 mmol), copper acetate (944 mg, 4.72 mmol), pyridine (4.5 mL) were added to 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 to obtain the corresponding compound 1i with a yield of 73-92%.

[0128] Step 6, 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. The mixture was reacted 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, 21.2×250mm, 5μm; elution system: methanol / water (volume ratio) = 10%-100%, containing 0.5‰ trifluoroacetic acid by volume, flow rate: 10ml / min, elution time: 40min) to obtain the corresponding compounds II-1 and II-2 with yields 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 (Bidec Pharmaceuticals, CAS No.: 38767-72-5).

[0131] Single configuration compound 2-1: 1 H NMR (600 MHz, methanol-d 4) 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 - d 4 ) 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 - d 4 ) 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 - d 4)δ165.5, 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 (Bidec Pharmaceuticals, CAS No.: 21901-41-7).

[0134] Single configuration compound 3-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.95 (s, 1H), 7.46 (d, J=8.7Hz, 1H), 7.26 (t, J=7.6Hz, 2H), 7.05 (d, J=8.7Hz, 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=11.5Hz, 1H), 1.45-1.27 (m, 3H). 13 C NMR (300MHz, methanol-d 4 )δ159.5, 159.4, 144.2, 138.2, 133.9, 133.9, 131.6, 130.4, 124.9, 122.7, 121.4, 119.1, 1 18.8, 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-d 4)δ7.81 (s, 1H), 7.34 (d, J = 9.2Hz, 1H), 7.27-7.22 (m, 1H), 6.91-6.76 (m, 5H), 6.65 (s, 1H), 6.52 (d, J = 5.5Hz, H), 4.42 (d, J = 12.8Hz, 1H) , 4.24 (d, J=12.8Hz, 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 (300MHz, methanol-d 4 )δ159.4, 159.2, 142.9, 142.9, 136.9, 133.2, 132.3, 131.6, 131.4, 125.1, 123.4, 123.0, 1 21.1, 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 (Bidec Pharmaceuticals, CAS No.: 13466-41-6).

[0137] Single configuration compound 4-1: 1 H NMR (600 MHz, methanol-d 4 )δ8.00 (s, 1H), 7.60 (t, J=7.3Hz, 1H), 7.57-7.54 (m, 2H), 7.51 (s, 1H), 7.49-7.47 (m, 2H), 7 .42 (d, J=8.2Hz, 1H), 7.23 (d, J=8.2Hz, 1H), 6.48 (s, 1H), 6.40 (d, J=7.3Hz, 1H), 4.32 (d, J=1 2.8Hz, 1H), 4.26 (d, J=12.8Hz, 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 (300MHz, methanol-d 4)δ164.4, 155.8, 142.4, 139.4, 139.4, 138.5, 134.4, 133.8, 131.6, 131.3, 129.6, 128.8, 1 24.6, 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-d 4 )δ7.93 (s, 1H), 7.56 (t, J=7.3Hz, 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, 1.8Hz, 1H), 4.32 (d, J =12.8Hz, 1H), 4.26 (d, J = 12.8Hz, 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). 13 C NMR (300MHz, methanol-d 4 )δ164.4, 155.8, 142.4, 138.5, 138.5, 138.3, 133.9, 133.7, 131.6, 131.4, 129.7, 128.8, 1 24.8, 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 (Bidec Pharmaceuticals, CAS No.: 142-08-5).

[0140] Single configuration compound 5-1: 1 H NMR (600 MHz, methanol-d 4)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-d 4 )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: 1 H NMR (600 MHz, methanol-d 4 )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-d 4)δ164.5, 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+1].

[0142] Single-configuration compounds 6-1 and 6-2 were prepared from 2-oxo-1,2-dihydropyridine-4-carbonitrile 1c (Bidec Pharmaceuticals, CAS No.: 94805-51-3).

[0143] Single configuration compound 6-1: 1 H NMR (600 MHz, methanol-d 4 )δ8.14 (s, 1H), 7.76 (d, J=6.9Hz, 1H), 7.71 (dd, J=9.2, 0.9Hz, 1H), 7.61-7.55 (m, 3H), 7.45 (dt, J= 6.9, 1.8Hz, 1H), 7.42 (d, J=9.2Hz, 1H), 7.06 (d, J=1.8Hz, 1H), 6.62 (dd, J=6.9, 1.8Hz, 1H), 4.34 (d, J=13.3Hz, 1H), 4.26 (d, J=13.3Hz, 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=11.9Hz, 1H), 1.56-1.45 (m, 3H). 13 C NMR (300MHz, methanol-d 4 )δ162.4, 141.8, 141.7, 141.0, 140.8, 135.2, 134.0, 131.8, 131.7, 129.3, 128.6, 127.7, 1 26.5, 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-d 4)δ7.84 (s, 1H), 7.70 (d, J=6.9Hz, 1H), 7.58-7.52 (m, 3H), 7.46 (dt, J=7.3, 1.8Hz, 1H), 7.37 (d, J=9.6Hz, 1H), 7.05 (d, J=1.4Hz, 1H), 7.00 (d, J=9.2Hz, 1H), 6.57 (dd, J=6.9, 1 .8Hz, 1H), 4.30 (d, J=13.3Hz, 1H), 4.26 (d, J=13.3Hz, 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 (300MHz, methanol-d 4 )δ162.5, 141.8, 141.8, 138.3, 138.1, 133.9, 133.4, 132.0, 131.7, 129.4, 128.6, 127.8, 1 26.5, 124.8, 121.5, 116.6, 112.6, 107.6, 101.8, 54.4, 52.5, 49.6, 32.2, 29.6, 29.1, 19.9. MSm / z(ESI): 414.2[M+1].

[0145] Single-configuration compounds 7-1 and 7-2 were prepared from 2-oxo-1,2-dihydropyridine-4-amide 1c (Bidec Pharmaceuticals, CAS No.: 175277-69-7).

[0146] Single configuration compound 7-1: 1 H NMR (600 MHz, methanol-d 4)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 - d 4 )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 - d 4 )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, lH), 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). 13 C NMR (300 MHz, methanol - d4 )δ169.0, 164.3, 147.8, 142.2, 140.2, 139.3, 138.2, 134.4, 133.7, 131.8, 131.7, 129.6, 1 28.8, 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 (Bidec Pharmaceuticals, CAS No.: 1003-56-1).

[0149] Single configuration compound 8-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.98 (s, 1H), 7.60 (t, J=7.8Hz, 1H), 7.57-7.54 (m, 2H), 7.53-7.51 (m, 2H), 7.46-7 .42 (m, 2H), 7.22 (d, J=9.2Hz, 1H), 6.42 (t, J=6.9Hz, 1H), 4.33 (d, J=13.3Hz, 1H), 4. 26(d, J=13.3Hz, 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 (300MHz, methanol-d 4 )δ164.6, 143.0, 140.6, 140.2, 137.0, 135.0, 133.7, 131.6, 131.2, 131.1, 129.7, 128.8, 1 24.3, 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: 1 H NMR (600 MHz, methanol-d 4)δ7.87(d, J=0.9Hz, 1H), 7.55(t, J=7.8Hz, 1H), 3.52-3.49(m, 3H), 7.44-7.42(m, 1H), 7.42-7.39 (m, 2H), 7.05 (d, J=8.7Hz, 1H), 6.38 (t, J=6.9Hz, 1H), 4.30 (d, J=1 3.3Hz, 1H), 4.26 (d, J=13.3Hz, 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 (300MHz, methanol-d 4 )δ164.6, 143.0, 140.2, 140.2, 139.2, 137.0, 134.3, 133.5, 131.6, 131.4, 131.1, 129.8, 1 28.9, 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 (Bidec Pharmaceuticals, CAS No.: 1003-68-5).

[0152] Single configuration compound 9-1: 1 H NMR (600 MHz, methanol-d 4)δ8.09(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 - d 4 )δ163.7, 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 - d 4 )δ8.04(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). 13 C NMR(300 MHz, methanol - d 4)δ163.7, 145.8, 142.6, 142.7, 139.5, 137.0, 134.5, 133.6, 131.6, 131.5, 129.8, 128.9, 1 24.5, 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 (Bidec Pharmaceuticals, CAS No.: 96530-75-5).

[0155] Single configuration compound 10-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.97 (s, 1H), 7.72 (t, J=7.3Hz, 1H), 7.61 (t, J=7.8Hz, 1H), 7.58 (dt, J=7.8, 1.4Hz, 1H), 7.54-7.52 (m, 2H), 7.44 (dt, J=7.3, 1.4Hz, 1H), 7.19 (d, J=8.7Hz, 1H), 7.47 (m, 1H), 7.33 (dd, J=10.6, 2.8Hz, 1H) , 4.33 (d, J = 13.3Hz, 1H), 4.27 (d, J = 13.3Hz, 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 (300MHz, methanol-d 4 )δ172.8 (d, J=267.3Hz), 165.7 (d, J=20.2Hz), 142.2 (d, J=14.4Hz), 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=17.3Hz), 100.9 (d, J=28.9Hz), 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-d 4 )δ8.02 (s, 1H), 7.71 (t, J=7.8Hz, 1H), 7.60-7.54 (m, 4H), 7.47-7.45 (m, 1H), 7.24 (d, J=8.7Hz, 1H), 6.47-6.44 (m, 1H), 7.33 (dd, J=7.8, 2.8Hz, 1H), 4.35 (d, J=13.3H z, 1H), 4.28 (d, J=13.3Hz, 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 (300MHz, methanol-d 4 ) δ172.8 (d, J=268.8Hz), 165.8 (d, J=18.8Hz), 142.2 (d, J=13.0Hz), 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=17.3Hz), 101.0 (d, J=27.4Hz), 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(1H)-one 1c (Bidec Pharmaceuticals, CAS No.: 37529-91-2).

[0158] Single configuration compound 11-1: 1 H NMR (600 MHz, methanol-d 4) 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 - d 4 ) 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 - d 4 ) 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 (300MHz, methanol-d 4 )δ164.7, 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(1H)-one 1c (Bidec Pharmaceuticals, CAS No.: 50650-59-4).

[0161] Single configuration compound 13-1: 1 H NMR (600 MHz, methanol-d 4 )δ7.90 (s, 1H), 7.81 (d, J=7.3Hz, 1H), 7.64 (t, J=7.8Hz, 1H), 7.61-7.59 (m, 1H), 7.58 (t, J=1.8Hz, 1H), 7. 51-7.49 (m, 1H), 7.46 (d, J=8.7Hz, 1H), 7.10 (d, J=8.7Hz, 1H), 6.94 (s, 1H), 6.64 (dd, J=6.9, 1.8Hz, 1H), 4 .34 (d, J=13.3Hz), 4.29 (d, J=13.3Hz), 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 (300MHz, methanol-d 4 )δ163.3, 143.5 (q, J=30.3Hz), 142.0, 141.8, 138.3, 138.0, 134.1, 133.9, 131.8, 131.8, 129.4, 128.7, 124.8, 12 3.4 (q, J=283.2Hz), 121.5, 119.4 (d, J=4.3Hz), 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-d 4 )δ7.86 (s, 1H), 7.73 (d, J = 7.3Hz), 7.55 (t, J = 7.8Hz), 7.53-7.51 (m, 2H), 7.47 (dt, J = 7.3, 1.8Hz , 1H), 7.33 (d, J=8.7Hz, 1H), 6.99 (d, J=8.7Hz, 1H), 6.91 (s, 1H), 6.57 (dd, J=6.9, 2.3Hz, 1H), 4.2 9(d, J=13.8Hz, 1H), 4.24(d, J=13.8Hz), 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 (300MHz, methanol-d 4 ) δ163.3, 143.7 (q, J=33.2Hz), 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=245.6Hz), 121.1, 119.4 (d, J=2.9Hz), 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 (Bidec Pharmaceuticals, CAS No.:

[0164] 19006-81-6).

[0165] Single configuration compound 14-1: 1 H NMR (600 MHz, methanol-d 4)δ7.89 (s, 1H), 7.74 (dd, J=7.3, 1.8Hz, 2H), 7.68 (d, J=7.3Hz, 1H), 7.64 (t, J=7.79, 1H), 7.60-7.58 (m, 2H), 7. 52-7.49 (m, 4H), 7.45 (d, J = 8.7Hz, 1H), 7.10 (d, J = 8.7Hz, 1H), 6.89 (d, J = 1.8Hz, 1H), 6.85 (dd, J = 7.3, 2.3Hz, 1H ), 4.35 (d, J = 12.8Hz, 1H), 4.30 (d, J = 12.8Hz, 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), 2.05-2.01m, 1H), 1.57-1.51 (m, 1H), 1.49-1.39 (m, 2H), 1.37-1.30 (m, 1H). 13 C NMR (300MHz, methanol-d 4 )δ164.6, 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: 1 H NMR (600 MHz, methanol-d 4)δ7.83 (s, 1H), 7.73-7.71 (m, 2H), 7.61 (d, J=6.9Hz, 1H), 7.58-7.56 (m, 2H), 7.53-7.49 (m, 5H), 7.3 6 (dd, J=8.7, 0.9Hz, 1H), 7.00 (d, J=8.7Hz, 1H), 6.88 (d, J=1.8Hz, 1H), 6.79 (d, J=7.3, 1.8Hz, 1H), 4 .31(d, J=13.3Hz, 1H), 4.27(d, J=13.3Hz, 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 (300MHz, methanol-d 4 )δ164.6, 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 disclosed compounds on ROCK2 kinase activity. The experimental method is briefly described as follows: The ROCK2 kinase inhibitory activity is determined using the HTRF KinEASE kit (PerkinElmer, Catalog No.: 62ST2PEB) based on time-resolved fluorescence technology. KD025 (MCE, CAS No.: 911417-87-3) is used as a positive control, and 3 replicate wells are set up in parallel. 4 μL of the test compound or buffer, 2 μL of the reaction substrate S are added to each well of a white 384-well plate. 2, 2μL ROCK2 kinase and 2μL ATP. Incubate in a 37℃ incubator for 30min. Add 5μL streptokinase labeled XL-665 and 5μL EuK labeled anti-phosphorylated protein kinase antibody in turn, react at room temperature for 60min, and use Tecan multifunctional microplate reader to detect fluorescence signal. The fluorescence intensity value of the sample well measured at λ=665nm is F1, and the fluorescence intensity value at λ=620nm is F2. The signal ratio calculation formula is: signal ratio = F1 / F2×10 4 The inhibitory rate of the test compound on protein kinase was calculated as follows: Inhibition rate (%) = [1-(test compound signal ratio-minimum signal ratio) / (maximum signal ratio-minimum signal ratio)] × 100. The maximum signal ratio is the solvent control, and the minimum signal ratio is the signal ratio of the XL-665 well without streptokinase labeling. The IC value of the inhibitory activity of the compound was calculated using GraphPad Prism8 software. 50 See Table 1 for values.

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

[0170] Compound <![CDATA[IC 50 (nM)]]> Compound <![CDATA[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 disclosed compounds have good activity in inhibiting ROCK2 kinase.

[0172] Test Example 2

[0173] The inhibitory effect of the disclosed compounds on ROCK1 kinase activity was determined. The experimental method was the same as that of Detection Example 1, but the incubation time was changed to 20 min. The detection results are shown in Table 2.

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

[0175] Compound <![CDATA[IC 50 (nM)]]> Compound <![CDATA[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 disclosed compounds have good activity in inhibiting ROCK1 kinase.

Claims

1. An indazole derivative or a pharmaceutically acceptable salt thereof, characterized in that: The structural formula of the indazole derivative is shown in general formula (I), in: G is a CH or N atom; Ring A is selected from 5- to 7-membered aryl, heteroaryl, cycloalkyl and heterocyclyl; Each R 1 are the same or different and are each independently selected from hydrogen, halogen, an alkyl group having 1 to 6 carbon atoms, a haloalkyl group having 1 to 6 carbon atoms, a haloalkoxy group having 1 to 6 carbon atoms, a cyano group, an amino group, a hydroxyl group, a nitro group, a carboxyl group, a hydroxyalkyl group having 1 to 6 carbon atoms, a cycloalkyl group, a heterocyclic group, an aryl group and a heteroaryl group; Each R 2 are the same or different and are each independently selected from hydrogen, halogen, cyano, hydroxyl, nitro, carboxyl, and hydroxyalkyl having 1 to 6 carbon atoms; Each R 3 are the same or different and are each independently selected from hydrogen, halogen, an alkyl group having 1 to 6 carbon atoms, a haloalkyl group having 1 to 6 carbon atoms, a haloalkoxy group having 1 to 6 carbon atoms, a cyano group, a hydroxyl group, a nitro group, a carboxyl group, and a hydroxyalkyl group having 1 to 6 carbon atoms; a is 0, 1, 2, 3 or 4; q is 0, 1, or 2; s is 0, 1, or 2.

2. The indazole derivative or pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The structural formula of the indazole derivative is shown in general formula (I-1), (I-2), (I-3) or (I-4), 3. The indazole derivative or pharmaceutically acceptable salt thereof according to claim 1, characterized in that: Ring A is selected from a 5- to 7-membered cycloalkyl group, a benzene ring and a heterocyclic group.

4. The indazole derivative or pharmaceutically acceptable salt thereof according to claim 3, characterized in that: Ring A is selected from cyclopentane, cyclohexane, cycloheptane, benzene ring and pyridine ring. The structural formula of the indazole derivative is shown in general formula (II-1), (II-2) or (II-3). Where n is 0, 1 or 2.

5. The indazole derivative or pharmaceutically acceptable salt thereof according to claim 4, characterized in that: The structural formula of the indazole derivative is shown in general formula (III-1), (III-2), (III-3), (III-4), (III-5), (III-6) or (III-7), 6. The indazole derivative or pharmaceutically acceptable salt thereof according to claim 5, characterized in that: The indazole derivative is any one of the following compounds:

7. The use of the indazole derivative or the pharmaceutically acceptable salt thereof according to claim 6, characterized in that: Any of the following: (1) Application in the preparation of ROCK kinase inhibitors; (2) Use in the preparation of a medicament for treating and / or preventing a disease or condition selected from inflammatory diseases, autoimmune diseases, fibrotic diseases, transplant rejection, diseases associated with 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 dermatological diseases.

8. The use according to claim 7, characterized in that: 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, Sjögren's syndrome, multiple sclerosis, dry eye disease, type I diabetes and complications associated therewith, atopic eczema, thyroiditis, contact dermatitis, Sjögren's syndrome and amyotrophic lateral sclerosis.

9. A drug, characterized in that The active ingredient is the indazole derivative or a pharmaceutically acceptable salt thereof as described in claim 6.

10. The method for preparing the indazole derivative or the pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The reaction formula is: The compounds represented by the general formula (IA) and the general formula (IV-1) or their salts undergo a reductive amination reaction to obtain the compound represented by the general formula (I) or its pharmaceutically acceptable salt.

11. The preparation method according to claim 10, characterized in that: The preparation method of general formula (IA) is: The compound represented by the general formula (IA-1) and the compound represented by the general formula (IA-2) or a salt thereof undergoes a coupling reaction to obtain the compound represented by the general formula (IA) or a salt thereof.

12. The preparation method according to claim 10, characterized in that: The preparation method of general formula (IV-1) is: The compound represented by the general formula (V-1) or its salt undergoes a deprotection reaction to obtain the compound represented by the general formula (IV-1) or its salt.

13. The preparation method according to claim 12, characterized in that: The preparation method of general formula (IV-1) is: in: The compounds represented by the general formula (IV-2) and the general formula (IV-4) or their salts undergo reductive amination reaction to obtain the compound represented by the general formula (V-1) or its salt; X is a bromine atom, a chlorine atom or an iodine atom; The compounds represented by the general formula (IV-3) and the general formula (IV-4) or their salts undergo coupling reaction to obtain the compound represented by the general formula (V-1) or its salt.

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