Pyrimido[6,1-a]isoquinolin-4-one derivatives and uses thereof

Abstract

The present invention relates to a pyrimido[6,1-a]isoquinolin-4-one derivative and its use. The structural formula of the pyrimido[6,1-a]isoquinolin-4-one derivative of the present invention is shown in Formula (I): #imgabs0# wherein R1, R2, R3, X, and Y are as described in the specification. The compound of the present invention can simultaneously inhibit PDE3 and PDE4 and exhibits superior efficacy compared to existing dual-target PDE3 and PDE4 inhibitors.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a pyrimido[6,1-a]isoquinolin-4-one derivative and a use thereof. Background Art

[0002] PDE3 and PDE4 belong to the PDE (Phosphodiesteras, phosphodiesterase) enzyme superfamily and are the main cAMP hydrolases in lung tissue cells. Among them, PDE3 can hydrolyze both cAMP and cGMP, and its ability to hydrolyze cAMP is about ten times that of cGMP. Inhibition of PDE3 blocks the intracellular cAMP hydrolysis process, and the accumulated cAMP can cause Ca2+ to accumulate on the sarcoplasmic reticulum. 2+ Large amounts of cAMP are released, promoting bronchial smooth muscle relaxation. PDE4 specifically hydrolyzes cAMP, playing a major regulatory role in the expression of pro- and anti-inflammatory mediators. Its inhibition can significantly reduce the release of harmful mediators from inflammatory cells. The dual-target inhibition strategy can synergistically increase intracellular cAMP / cGMP concentrations, activate downstream signaling pathways, and achieve dual enhancement of airway relaxation and anti-inflammatory effects, while also promoting airway clearance.

[0003] WO2000058308A1 discloses a PDE3 / 4 dual-target inhibitor RPL554, which has positive clinical results.

[0004]

[0005] However, research data of RPL554 show that its inhibitory activity against PDE4 is significantly insufficient, while its inhibitory activity against PDE3 still has room for improvement. Summary of the Invention

[0006] To address unmet clinical needs, it is necessary to develop more efficient and safe PDE3 / 4 dual-target inhibitors. To this end, the present invention provides a pyrimido[6,1-a]isoquinolin-4-one derivative and its use.

[0007] Therefore, one object of the present invention is to provide a pyrimido[6,1-a]isoquinolin-4-one derivative as a PDE3 / 4 dual-target inhibitor.

[0008] Another object of the present invention is to provide a pharmaceutical composition comprising the above compound.

[0009] The technical solution of the present invention is achieved by adopting the following solution.

[0010] In one aspect, the present invention provides a compound represented by general formula (I) or a stereoisomer, tautomer, deuterated substance, prodrug or a pharmaceutically acceptable salt thereof:

[0011]

[0012] in,

[0013] R1 is selected from -NR4R5 or cyclic Cy;

[0014] R4 and R5 are independently selected from H, C1-C6 alkyl, and C3-C6 cycloalkyl;

[0015] Ring Cy is selected from 4-7 membered heteroaryl and aryl groups containing 1-5 (e.g., 1, 2, 3, 4 or 5) heteroatoms selected from N, O, and S, wherein the 4-7 membered heteroaryl and aryl groups containing 1-5 heteroatoms selected from N, O, and S are optionally substituted by 0, 1, 2, 3, 4 or 5 R 1a replace;

[0016] Each R 1a are the same or different and are each independently selected from H, deuterium, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C3-C6 cycloalkoxy, hydroxy, cyano, amino and thiol;

[0017] R2 is selected from H, hydroxy, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C3-C6 cycloalkoxy, and the C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C3-C6 cycloalkoxy is optionally substituted with 0, 1, 2 or 3 substituents selected from halogen, hydroxy, cyano, amino and mercapto;

[0018] R3 is selected from H, deuterium, C1-C6 alkyl, C3-C6 cycloalkyl, wherein the C1-C6 alkyl, C3-C6 cycloalkyl is optionally substituted with 0, 1, 2 or 3 substituents selected from deuterium, halogen, hydroxy, cyano, amino and thiol;

[0019] X is -(CH2) m -; m is 1, 2, 3 or 4; Y is -(CH2) n -; n is 1, 2, 3 or 4.

[0020] Preferably, the compound has a structure represented by formula (II), formula (III) or formula (IV):

[0021]

[0022] Preferably, R1 is -NR4R5 or cycloCy, when it is -NR4R5, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; when it is cycloCy, cycloCy is selected from 4-6 membered heteroaryl and aryl groups containing 1-4 (e.g., 1, 2, 3 or 4) heteroatoms selected from N, O, and S, and the 4-6 membered heteroaryl and aryl groups containing 1-4 heteroatoms selected from N, O, and S are optionally replaced by 0, 1, 2, 3 or 4 R 1a replace;

[0023] Preferably, R1 is -NR4R5 or cycloCy, when it is -NR4R5, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; when it is cycloCy, cycloCy is selected from 4-6 membered heteroaryl and aryl groups containing 1-3 (e.g., 1, 2 or 3) heteroatoms selected from N, O, and S, and the 4-6 membered heteroaryl and aryl groups containing 1-3 heteroatoms selected from N, O, and S are optionally replaced by 0, 1, 2 or 3 R 1a replace;

[0024] Preferably, R1 is -NR4R5 or cycloCy, when it is -NR4R5, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; when it is cycloCy, cycloCy is selected from 4-6 membered heteroaryl and aryl groups containing 1-3 (e.g., 1, 2 or 3) heteroatoms selected from N, O, and S, and the 4-6 membered heteroaryl and aryl groups containing 1-3 heteroatoms selected from N, O, and S are optionally replaced by 0, 1 or 2 R 1a replace;

[0025] Preferably, R1 is -NR4R5 or cycloCy, when it is -NR4R5, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; when it is cycloCy, cycloCy is selected from 5-6 membered heteroaryl and aryl groups containing 1-3 (e.g., 1, 2 or 3) heteroatoms selected from N, O, and S, and the 5-6 membered heteroaryl and aryl groups containing 1-3 heteroatoms selected from N, O, and S are optionally replaced by 0, 1, 2 or 3 R 1a replace;

[0026] Preferably, R1 is -NR4R5 or cycloCy, when it is -NR4R5, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; when it is cycloCy, cycloCy is selected from 5-6 membered heteroaryl and aryl containing 1-3 (e.g., 1, 2 or 3) N heteroatoms, and the 5-6 membered heteroaryl and aryl containing 1-3 N heteroatoms are optionally replaced by 0, 1 or 2 R 1a replace.

[0027] Preferably, R1 is -NR4R5 or cycloCy, when it is -NR4R5, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; when it is cycloCy, cycloCy is selected from

[0028]

[0029] Where w is 0, 1 or 2, v is 0, 1, 2, 3, 4 or 5, and u is 0, 1, 2 or 3;

[0030] Preferably, R 1a Each is independently selected from H, deuterium, halogen, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy, hydroxy, cyano, amino and thiol;

[0031] Preferably, R 1a are each independently selected from H, deuterium, halogen, C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy and hydroxy;

[0032] Preferably, R 1a are each independently selected from H, deuterium, halogen, C1-C3 alkyl, C3-C4 cycloalkyl, C1-C3 alkoxy, C3-C4 cycloalkoxy and hydroxy;

[0033] Preferably, R 1a Each is independently selected from H, deuterium, F, Cl, Br, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy and hydroxy;

[0034] Preferably, R 1a Each is independently selected from H, deuterium, F, Cl, Br, C1-C3 alkyl, C3-C4 cycloalkyl, C1-C3 alkoxy, C3-C4 cycloalkoxy and hydroxy;

[0035] Preferably, R 1a are each independently selected from H, deuterium, F, Cl, Br, methyl, ethyl, propyl, cyclopropyl, cyclobutyl and hydroxyl;

[0036] Preferably, R1 is -NR4R5 or cycloCy. When it is -NR4R5, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; when it is cycloCy, cycloCy is selected from:

[0037]

[0038] Preferably, R2 is selected from H, hydroxy, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy, and the C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy are optionally substituted with 0, 1, 2 or 3 substituents selected from halogen, hydroxy, cyano, amino and mercapto;

[0039] Preferably, R2 is selected from C1-C3 alkyl, C3-C4 cycloalkyl, C1-C3 alkoxy, C3-C4 cycloalkoxy, and the C1-C3 alkyl, C3-C4 cycloalkyl, C1-C3 alkoxy, C3-C4 cycloalkoxy are optionally substituted with 0, 1, 2 or 3 substituents selected from halogen, hydroxy, cyano, amino and mercapto.

[0040] Preferably, R2 is selected from methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy, ethoxy, propoxy, and the methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy, ethoxy, propoxy are optionally substituted with 0, 1, 2 or 3 substituents selected from halogen, hydroxy, cyano, amino and thiol.

[0041] Preferably, R3 is selected from H, deuterium, C1-C4 alkyl, C3-C5 cycloalkyl, wherein the C1-C4 alkyl, C3-C5 cycloalkyl is optionally substituted with 0, 1, 2 or 3 substituents selected from deuterium, halogen, hydroxyl, cyano, amino and thiol;

[0042] Preferably, R3 is selected from C1-C3 alkyl, C3-C4 cycloalkyl, and the C1-C3 alkyl, C3-C4 cycloalkyl is optionally substituted with 0, 1, 2 or 3 substituents selected from deuterium, halogen, hydroxyl, cyano, amino and mercapto;

[0043] Preferably, R3 is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl are optionally substituted with 0, 1, 2 or 3 substituents selected from deuterium, halogen, hydroxyl, cyano, amino and thiol.

[0044] Preferably, X is -(CH2) m -, m is 1, 2 or 3;

[0045] Preferably, X is -(CH2) m -, m is 1 or 2.

[0046] Preferably, X is -(CH2) m -, m is 1.

[0047] Preferably, Y is -(CH2) n -, n is 1, 2 or 3;

[0048] Preferably, Y is -(CH2) n -, n is 1 or 2.

[0049] In a specific embodiment, in formula (I), formula (II), formula (III) or formula (IV), R1 is -NR4R5 or cycloCy, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; cycloCy is selected from 5-6 membered heteroaryl and aryl containing 1-3 (e.g., 1, 2 or 3) N heteroatoms, and the 5-6 membered heteroaryl and aryl containing 1-3 N heteroatoms are optionally replaced by 0, 1 or 2 R 1a replace;

[0050] R 1a Each is independently selected from H, deuterium, halogen, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy, hydroxy, cyano, amino and thiol;

[0051] R2 is selected from H, hydroxy, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy, and the C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy is optionally substituted with 0, 1, 2 or 3 substituents selected from halogen, hydroxy, cyano, amino and mercapto;

[0052] R3 is selected from H, deuterium, C1-C4 alkyl, C3-C5 cycloalkyl, wherein the C1-C4 alkyl, C3-C5 cycloalkyl is optionally substituted with 0, 1, 2 or 3 substituents selected from deuterium, halogen, hydroxy, cyano, amino and mercapto;

[0053] X is -(CH2) m -, m is 1, 2 or 3;

[0054] Y is -(CH2) n -, n is 1, 2 or 3. In another specific embodiment, in formula (I), formula (II), formula (III) or formula (IV), R1 is -NR4R5 or cycloCy, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; Cy is selected from

[0055]

[0056] Where w is 0, 1 or 2, v is 0, 1, 2, 3, 4 or 5, and u is 0, 1, 2 or 3;

[0057] R 1aEach is independently selected from H, deuterium, halogen, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy, hydroxy, cyano, amino and thiol;

[0058] R2 is selected from H, hydroxy, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy, and the C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy is optionally substituted with 0, 1, 2 or 3 substituents selected from halogen, hydroxy, cyano, amino and mercapto;

[0059] R3 is selected from H, deuterium, C1-C4 alkyl, C3-C5 cycloalkyl, wherein the C1-C4 alkyl, C3-C5 cycloalkyl is optionally substituted with 0, 1, 2 or 3 substituents selected from deuterium, halogen, hydroxy, cyano, amino and mercapto;

[0060] X is -(CH2) m -, m is 1, 2 or 3;

[0061] Y is -(CH2) n -, n is 1, 2 or 3.

[0062] In another specific embodiment, in formula (I), formula (II), formula (III) or formula (IV), R1 is -NR4R5 or cycloCy, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; cycloCy is selected from

[0063]

[0064] Where w is 0, 1 or 2, v is 0, 1, 2, 3, 4 or 5, and u is 0, 1, 2 or 3;

[0065] R 1a are each independently selected from H, deuterium, halogen, C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy and hydroxy;

[0066] R2 is selected from C1-C3 alkyl, C3-C4 cycloalkyl, C1-C3 alkoxy, C3-C4 cycloalkoxy, and the C1-C3 alkyl, C3-C4 cycloalkyl, C1-C3 alkoxy, C3-C4 cycloalkoxy is optionally substituted with 0, 1, 2 or 3 substituents selected from halogen, hydroxy, cyano, amino and mercapto;

[0067] R3 is selected from C1-C3 alkyl, C3-C4 cycloalkyl, and the C1-C3 alkyl, C3-C4 cycloalkyl is optionally substituted with 0, 1, 2 or 3 substituents selected from deuterium, halogen, hydroxy, cyano, amino and mercapto;

[0068] X is -(CH2) m -, m is 1, 2 or 3;

[0069] Y is -(CH2) n -, n is 1, 2 or 3.

[0070] In another embodiment, in Formula (I), Formula (II), Formula (III) or Formula (IV), R1 is -NR4R5 or cycloCy, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; cycloCy is selected from:

[0071]

[0072] R2 is selected from H, hydroxy, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy, and the C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, C3-C5 cycloalkoxy is optionally substituted with 0, 1, 2 or 3 substituents selected from halogen, hydroxy, cyano, amino and mercapto;

[0073] R3 is selected from H, deuterium, C1-C4 alkyl, C3-C5 cycloalkyl, wherein the C1-C4 alkyl, C3-C5 cycloalkyl is optionally substituted with 0, 1, 2 or 3 substituents selected from deuterium, halogen, hydroxy, cyano, amino and mercapto;

[0074] X is -(CH2) m -, m is 1, 2 or 3;

[0075] Y is -(CH2) n -, n is 1, 2 or 3.

[0076] In another embodiment, in Formula (I), Formula (II), Formula (III) or Formula (IV), R1 is -NR4R5 or cycloCy, R4 and R5 are independently selected from H, C1-C4 alkyl, C3-C5 cycloalkyl; and cycloCy is selected from:

[0077]

[0078] R2 is selected from C1-C3 alkyl, C3-C4 cycloalkyl, C1-C3 alkoxy, C3-C4 cycloalkoxy, and the C1-C3 alkyl, C3-C4 cycloalkyl, C1-C3 alkoxy, C3-C4 cycloalkoxy is optionally substituted with 0, 1, 2 or 3 substituents selected from halogen, hydroxy, cyano, amino and mercapto;

[0079] R3 is selected from C1-C3 alkyl, C3-C4 cycloalkyl, and the C1-C3 alkyl, C3-C4 cycloalkyl is optionally substituted with 0, 1, 2 or 3 substituents selected from deuterium, halogen, hydroxy, cyano, amino and mercapto;

[0080] X is -(CH2) m -, m is 1, 2 or 3;

[0081] Y is -(CH2) n -, n is 1, 2 or 3.

[0082] In another embodiment, the compound represented by general formula (I) is selected from:

[0083]

[0084] The compound of formula (I) of the present invention can be prepared by chemical synthesis methods known in the art, for example, by retrosynthesis using commercially available raw materials. Specific synthesis examples are provided in the embodiments of the present invention.

[0085] In another aspect, the present invention provides a pharmaceutical composition comprising the above-mentioned compound or its stereoisomers, tautomers, deuterated substances, prodrugs or pharmaceutically acceptable salts and a pharmaceutically acceptable excipient, carrier or diluent.

[0086] Compared to the prior art, the compounds of the present invention, or their stereoisomers, tautomers, deuterated forms, prodrugs, or pharmaceutically acceptable salts, or pharmaceutical compositions, can simultaneously inhibit PDE3 and PDE4. Furthermore, compared to the existing PDE3 and PDE4 dual-target inhibitor RPL554, the compounds of the present invention, or their stereoisomers, tautomers, deuterated forms, prodrugs, or pharmaceutically acceptable salts, or pharmaceutical compositions, have comparable or superior inhibitory activity against PDE3, significantly greater inhibitory activity against PDE4 than RPL554, and exhibit stronger activity against TNF-α and cAMP.

[0087] definition

[0088] Unless otherwise specified, the meaning and scope of the terms of the present invention are explained below in an exemplary manner.

[0089] “ ” indicates the attachment site.

[0090] The minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, for example, the prefix (C a-b )alkyl represents any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, (C 1-6 ) Alkyl refers to an alkyl group containing 1 to 6 carbon atoms. The alkyl group is branched or linear.

[0091] Atoms described herein include their isotopes, for example, hydrogen can be deuterium or tritium.

[0092] "Alkyl" refers to a monovalent straight-chain or branched saturated hydrocarbon group, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl and other similar groups. 1-6 Alkyl. More preferably C 1-4 Alkyl. More preferably C 1-3 alkyl.

[0093] "Cycloalkyl" refers to a saturated monocyclic, linked, spirocyclic, fused, or bridged cycloalkyl group, which may be combined with other groups. Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Cycloalkyl groups with 3-6 members are preferred. Cycloalkyl groups with 3-5 members are preferred. Cycloalkyl groups with 3-4 members are preferred.

[0094] "Halogen" refers to fluorine, chlorine, bromine or iodine, with fluorine, chlorine and bromine being preferred.

[0095] "Aryl" refers to a substituted or unsubstituted monocyclic or polycyclic aromatic group, including but not limited to phenyl and naphthyl. A 6-10 membered monocyclic or bicyclic aromatic group is preferred. Phenyl or naphthyl are more preferred. Phenyl is most preferred.

[0096] "Heteroaryl" refers to a substituted or unsubstituted 5- or 6-membered monoheteroaromatic ring system, or a substituted or unsubstituted 9- or 10-membered fused or biheteroaromatic ring system containing 1-4 heteroatoms independently selected from N, O, or S, with the remaining ring atoms being carbon atoms. Examples of heteroaryl moieties include, but are not limited to, thienyl, furanyl, imidazolyl, isoxazolyl, oxazolyl, pyrazolyl, pyrrolyl, thiadiazolyl, triazolyl, pyridinyl, pyridazinyl, pyrimidinyl, indolyl, indazolyl, quinolinyl, isoquinolinyl, benzimidazolyl, or benzothiazolyl.

[0097] "Pharmaceutically acceptable salts" refer to conventional acid addition salts or base addition salts which retain the biological effectiveness and properties of the compound of formula (I) and are formed from suitable non-toxic organic or inorganic acids or organic or inorganic bases. Examples of acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid, and nitric acid, and those derived from organic acids. Examples of organic acids include acetic acid, propionic acid, glycolic acid, oxalic acid, stearic acid, ascorbic acid, p-toluenesulfonic acid, salicylic acid, methanesulfonic acid, ethanesulfonic acid, oxalic acid, succinic acid, citric acid, maleic acid, hydroxymaleic acid, lactic acid, fumaric acid, tartaric acid, malic acid, isethionic acid, benzenesulfonic acid, trifluoroacetic acid, mandelic acid, and the like. Examples of base addition salts include those derived from inorganic acids such as ammonium salts, calcium salts, iron salts, aluminum salts, sodium salts, potassium salts, zinc salts, and magnesium salts, and those derived from organic acids. The organic base includes salts of primary, secondary and tertiary amines, such as trimethylamine, triethylamine, tripropylamine, diethanolamine, ethylenediamine, ethanolamine and the like.

[0098] "Prodrug" refers to a prodrug that can be converted into a compound of the present invention and a pharmaceutically acceptable salt thereof in vivo. DETAILED DESCRIPTION

[0099] The technical solutions of the present invention are further described below with reference to specific examples. The examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention, and the embodiments of the present invention are not limited thereto. Any other changes, substitutions, modifications, simplifications, etc. that do not deviate from the technical ideas and method principles of the present invention shall be considered equivalent replacement methods and are included within the scope of protection of the present invention.

[0100] Unless otherwise specified, the chemicals and devices used in the following examples are commercially available.

[0101] Synthesis of intermediates

[0102]

[0103] 1. Synthesis of intermediate Int-A

[0104]

[0105] (1) Synthesis of A-2

[0106] A-1 (30 g, 166.48 mmol) was dissolved in acetic acid (200 mL), and ammonium acetate (32.1 g, 416.45 mmol) was added. o C for 15 minutes. Nitromethane (30.5 g, 499.67 mmol) was added dropwise to the reaction mixture and stirred at 100 o C and stirred for 4 hours. After the reaction was completed, the reaction solution was slowly poured into 500 mL of water. The precipitated solid was filtered and dried, then slurried with 100 mL of methanol and dried to obtain A-2 (21 g, yellow solid) with a yield of 57%.

[0107] MS m / z(ESI): 224.0[M+H] + .

[0108] (2) Synthesis of A-3

[0109] A-2 (21 g, 94.07 mmol) was dissolved in dimethyl sulfoxide (165 mL) and acetic acid (80 mL) at 0 o Sodium borohydride (5.34 g, 141.16 mmol) was added in portions at 10 oC. Stir for 1 hour. After the reaction is complete, the reaction solution is slowly poured into 500 mL of water and extracted with ethyl acetate. The combined organic phases are washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant is removed by filtration, and the filtrate is concentrated under reduced pressure to obtain A-3 (21 g, yellow gum) in a 99% yield.

[0110] MS m / z(ESI): 226.2[M+H] + .

[0111] 1 H NMR (400 MHz, Chloroform- d ) δ 6.82 (d, J = 8.2 Hz, 1H), 6.77 -6.66 (m, 2H), 4.58 (t, J = 7.2 Hz, 2H), 4.08 (q, J = 7.2 Hz, 2H), 3.85 (s,3H), 3.25 (t, J = 7.6 Hz, 2H), 1.46 (t, J = 7.2 Hz, 3H).

[0112] (3) Synthesis of A-4

[0113] A-3 (20 g, 88.79 mmol) was dissolved in methanol (100 mL), and 5% palladium carbon (4.44 g) was added under nitrogen atmosphere. The atmosphere was replaced with hydrogen three times, and the mixture was stirred at 40 °C under hydrogen (15 psi). o C for 16 hours. After the reaction was completed, palladium carbon was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain A-4 (17 g, colorless gum) with a yield of 98%.

[0114] MS m / z(ESI): 196.1 [M+H] + .

[0115] (4) Synthesis of A-5

[0116] A-4 (17 g, 87.06 mmol) was added to ethyl cyanoacetate (32 mL) and stirred at 100 °C under nitrogen atmosphere. o C. and stirred for 16 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and the resulting crude product was purified by flash silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1 to dichloromethane / methanol = 10 / 1) to afford A-5 (18 g, light yellow solid) in a 79% yield.

[0117] MS m / z(ESI): 263.1 [M+H] + .

[0118] 1 H NMR (400 MHz, Chloroform- d ) δ 6.83 (d, J = 7.6 Hz, 1H), 6.73 (d, J = 7.6 Hz, 2H), 6.10 (s, 1H), 4.10 (q, J = 7.2 Hz, 2H), 3.86 (s, 3H), 3.54 (q, J = 6.8 Hz, 2H), 3.33 (s, 2H), 2.79 (t, J = 7.2 Hz, 2H), 1.47 (t, J = 7.2 Hz, 3H).

[0119] (5) Synthesis of A-6

[0120] A-5 (18 g, 68.62 mmol) was added portionwise to 85 o C phosphorus oxychloride (180 mL), at 85 o The mixture was stirred at 40°C for 3 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure to remove most of the phosphorus oxychloride. The resulting crude product was diluted with dichloromethane, poured into ice water, and the pH was adjusted to 7 with saturated sodium bicarbonate aqueous solution. The product was extracted with dichloromethane. The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the intermediate A-6 (16 g, crude, yellow solid) in a 95% yield.

[0121] MS m / z(ESI): 245.2 [M+H] + .

[0122] (6) Synthesis of A-7

[0123] At 0 o C, A-6 (16.0 g, 65.50 mmol) was added to concentrated sulfuric acid (160 mL) in portions, and the mixture was stirred at 10 o The mixture was stirred at 4°C for 3 hours. After the reaction was complete, the reaction solution was slowly poured into ice water, and the pH was adjusted to 7 with 4 M aqueous sodium hydroxide solution. The mixture was extracted with dichloromethane. The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain A-7 (11.4 g, yellow solid) in a 66% yield.

[0124] MS m / z(ESI): 263.2 [M+H] + .

[0125] 1 H NMR (400 MHz, DMSO- d 6) δ 9.44 (t, J = 3.2 Hz, 1H), 7.06 (s, 1H), 6.83 (s, 1H), 6.26 (s, 2H), 5.07 (s, 1H), 4.06 (t, J = 7.2 Hz, 2H), 3.78 (s,3H), 3.27 (td, J = 6.4, 2.8 Hz, 2H), 2.72 (t, J = 6.4 Hz, 2H), 1.34 (t, J =7.2 Hz, 3H).

[0126] (7) Synthesis of A-8

[0127] At 0 o Sodium metal (5.81 g, 252.72 mmol) was added to ethanol (200 mL) in portions at 80 °C and stirred until all the sodium metal reacted. A-7 (11.4 g, 43.46 mmol) was added to the freshly prepared sodium ethoxide solution and stirred at 80 °C. o C for 1 hour, then diethyl carbonate (15.80 mL) was added to the reaction mixture and the mixture was stirred at 80 o C. and stirred for 8 hours. After the reaction was completed, the reaction solution was slowly poured into ice water and the pH was adjusted to 7 with 2 M hydrochloric acid. The solid precipitate was filtered and dried under reduced pressure to obtain A-8 (8.5 g, yellow solid) with a yield of 68%.

[0128] MS m / z(ESI): 289.1 [M+H] + .

[0129] 1 H NMR (400 MHz, DMSO- d 6) δ 11.20 (s, 1H), 7.35 (s, 1H), 6.94 (s, 1H), 6.22 (d, J = 2.1 Hz, 1H), 4.08 (q, J = 6.9 Hz, 2H), 3.89 (t, J= 6.2 Hz, 2H),3.83 (s, 3H), 2.88 (t, J = 6.2 Hz, 2H), 1.35 (t, J = 6.9 Hz, 3H).

[0130] (8) Synthesis of A-9

[0131] A-8 (8.0 g, 27.75 mmol) was added to phosphorus oxychloride (120 mL) and stirred at 100 o The mixture was stirred at 40°C for 3 hours. After the reaction was complete, the filtrate was concentrated under reduced pressure to remove most of the phosphorus oxychloride. The resulting crude product was diluted with dichloromethane, poured into ice water, and the pH was adjusted to 7 with saturated sodium bicarbonate aqueous solution. The product was extracted with dichloromethane. The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain A-9 (8.0 g, crude product, yellow solid) in a 94% yield.

[0132] MS m / z(ESI): 307.1 [M+H] + .

[0133] (9) Synthesis of intermediate Int-A

[0134] A-9 (1.5 g, 4.89 mmol) and A-10 (1.14 g, 7.54 mmol, purchased) were added to isopropanol (30 mL) and stirred at 90 o C. and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and the solid precipitate was filtered, washed with ethyl acetate, and dried under reduced pressure to obtain intermediate Int-A (2.0 g, yellow solid) with a yield of 97%.

[0135] MS m / z(ESI): 422.2 [M+H] + .

[0136] 2. Synthesis of intermediates Int-B~Int-D

[0137] Intermediates Int-B~Int-D were prepared by referring to the synthetic method of intermediate Int-A.

[0138]

[0139] Example 1: Synthesis of Compound 1

[0140]

[0141] Synthesis of compound 1-1

[0142] Compound Int-D (97.9 mg, 0.25 mmol), compound a (360.2 mg, 1.44 mmol), cesium carbonate (700.5 mg, 2.15 mmol) and sodium iodide (215.8 mg, 1.44 mmol) were added to butanone (10 mL) and the mixture was stirred at 110 °C under nitrogen protection. o C. and stirred for 16 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by flash silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to obtain compound 1-1 (34.1 mg, yellow oil) in a yield of 24%.

[0143] MS m / z(ESI): 561.4[M+H] + .

[0144] Synthesis of compound 1-2

[0145] Compound 1-1 (34.1 mg, 0.061 mmol) was dissolved in dichloromethane (2 mL), followed by the addition of trifluoroacetic acid (0.5 mL) and the mixture was stirred at 25 °C for 1 h. o C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain compound 1-2 (26.7 mg, yellow oil) with a yield of 95%.

[0146] MS m / z(ESI): 461.4[M+H] + .

[0147] Synthesis of compound 1

[0148] Compound b (2.3 mg, 0.02 mmol), HATU (15.2 mg, 0.04 mmol), DIPEA (10.3 mg, 0.08 mmol) and compound 1-2 (18.4 mg, 0.04 mmol) were added to DMF (1.5 mL) and stirred at 25 o The reaction was stirred at 4°C for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 1 (1.2 mg, yellow solid) in a 5% yield.

[0149] MS m / z(ESI): 556.5[M+H] + .

[0150] 1 H NMR (400 MHz, Chloroform-d ) δ 8.22 (s, 1H), 7.00 (d, J = 7.6 Hz,2H), 6.69 (d, J = 20.8 Hz, 2H), 5.42 (s, 1H), 4.64 (dd, J = 13.6, 5.2 Hz,2H), 4.34 (dd, J = 10.8, 5.7 Hz, 1H), 4.29 - 4.05 (m, 3H), 3.91 (s, 3H), 3.87(s, 1H), 3.78 (dd, J = 10.4, 5.6 Hz, 1H), 3.74 (s, 3H), 3.04 (s, 1H), 2.94(t, J = 6.4 Hz, 2H), 2.34 (s, 3H), 2.14 (d, J = 9.6 Hz, 6H), 1.25 (s, 1H).

[0151] Example 2: Synthesis of Compound 2

[0152]

[0153] Synthesis of compound 2

[0154] Compound c (7.3 mg, 0.065 mmol), HATU (36.9 mg, 0.097 mmol), DIPEA (42.0 mg, 0.325 mmol) and compound 1-2 (29.9 mg, 0.065 mmol) were added to DMF (1.5 mL) and the mixture was stirred at 25 ℃ for 1 h. o The reaction was stirred at 4°C for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 2 (2.2 mg, white solid) in a 6% yield.

[0155] MS m / z(ESI) = 555.3 [M+H] + .

[0156] 1 H NMR (400 MHz, Chloroform- d ) δ11.24 (s, 1H), 7.49 (s, 1H), 6.86 (s, 2H), 6.69 (d, J = 16.4 Hz, 2H), 6.61 (s, 1H), 5.46 (s, 1H), 4.59 (t, J =6.4 Hz, 4H), 4.32 (t, J = 9.6 Hz, 1H), 4.18 (dd, J = 10.4, 6.0 Hz, 1H), 4.08- 4.00 (m, 2H), 3.90 (s, 3H), 3.76 (s, 3H), 3.45 (p, J = 7.2 Hz, 1H), 2.91(t, J = 6.4 Hz, 2H), 2.27 (s, 3H), 2.03 (s, 6H).

[0157] Example 3: Synthesis of Compound 3

[0158]

[0159] Synthesis of compound 3-1

[0160] Compound Int-C (499 mg, 1.23 mmol), compound a (923 mg, 3.69 mmol), cesium carbonate (3.41 g, 10.47 mmol) and sodium iodide (1.05 g, 7.01 mmol) were added to butanone (10 mL) and heated at 110°C under nitrogen protection. o C. and stirred for 16 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by flash silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to obtain compound 3-1 (427 mg, yellow solid) in a 60% yield.

[0161] MS m / z(ESI) = 575.4 [M+H] + .

[0162] Synthesis of compound 3-2

[0163] Compound 3-1 (400 mg, 0.696 mmol) was dissolved in dichloromethane (10 mL), followed by the addition of trifluoroacetic acid (3 mL) and the reaction mixture was stirred at 25 o C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain compound 3-2 (300 mg, brown solid) with a yield of 91%.

[0164] MS m / z(ESI) = 475.3 [M+H] + .

[0165] Synthesis of compound 3

[0166] Compound b (6.8 mg, 0.06 mmol), HATU (34.2 mg, 0.09 mmol), DIPEA (24.6 mg, 0.19 mmol) and compound 3-2 (28.5 mg, 0.06 mmol) were added to DMF (2 mL) and stirred at 25 o The reaction was stirred at 4°C for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 3 (2.0 mg, white solid) in a 6% yield.

[0167] MS m / z(ESI): 570.4[M+H] + .

[0168] 1 H NMR (400 MHz, Methanol- d 4) δ 7.95 (s, 1H), 7.17 (d, J = 4.0 Hz,2H), 6.95 (s, 1H), 6.69 (s, 1H), 5.56 (s, 1H), 4.63 (dd, J = 13.6, 4.4 Hz,1H), 4.14 (q, J = 6.8 Hz, 4H), 3.83 - 3.60 (m, 9H), 3.03 (t, J = 6.4 Hz, 2H),2.37 (s, 3H), 2.28 (s, 3H), 2.24 (s, 3H), 1.42 (t, J = 7.2 Hz, 3H).

[0169] Example 4: Synthesis of Compound 4

[0170]

[0171] Synthesis of compound 4

[0172] Compound d (8.3 mg, 0.065 mmol), HATU (36.9 mg, 0.097 mmol), DIPEA (42.0 mg, 0.325 mmol) and compound 1-2 (29.9 mg, 0.065 mmol) were added to DMF (1 mL) and the mixture was stirred at 25 ℃ for 1 h. o The reaction was stirred at 4°C for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 4 (2.8 mg, white solid) in an 8% yield.

[0173] MS m / z(ESI) = 570.3 [M+H] + .

[0174] 1 H NMR (400 MHz, Chloroform- d ) δ 13.13 (s, 1H), 6.85 (d, J = 3.2 Hz,2H), 6.69 (d, J = 14.4 Hz, 2H), 5.47 (s, 1H), 4.73 - 4.62 (m, 2H), 4.62 -4.53 (m, 2H), 4.29 (t, J = 9.6 Hz, 1H), 4.18 (dd, J = 10.4, 5.6 Hz, 1H), 4.05(t, J = 6.4 Hz, 2H), 3.90 (s, 3H), 3.76 (s, 3H), 3.49 - 3.39 (m, 1H), 2.92(t, J = 6.4 Hz, 2H), 2.52 (s, 3H), 2.26 (s, 3H), 2.04 (d, J = 7.2 Hz, 6H).

[0175] Example 5: Synthesis of Compound 5

[0176]

[0177] Synthesis of compound 5-1

[0178] Compound Int-D (493 mg, 1.26 mmol), compound e (1.00 g, 3.79 mmol), cesium carbonate (3.58 g, 10.99 mmol) and sodium iodide (1.08 g, 7.21 mmol) were added to DMF (10 mL) and stirred at 110 °C under nitrogen protection. o C. and stirred for 16 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was extracted with water and ethyl acetate. The organic phase was concentrated under reduced pressure, and the resulting crude product was purified by flash silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to obtain compound 5-1 (136 mg, yellow oil) in a yield of 19%.

[0179] MS m / z(ESI) = 575.4 [M+H] + .

[0180] Synthesis of compound 5-2

[0181] Compound 5-1 (136 mg, 0.237 mmol) was dissolved in dichloromethane (3 mL), followed by the addition of trifluoroacetic acid (0.5 mL) and the mixture was stirred at 25 o C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain compound 5-2 (110 mg, yellow oil) with a yield of 98%.

[0182] MS m / z(ESI) = 475.3 [M+H] + .

[0183] Synthesis of compound 5

[0184] Compound b (19.0 mg, 0.168 mmol), HATU (120.2 mg, 0.316 mmol), DIPEA (135.7 mg, 1.050 mmol) and compound 5-2 (99.7 mg, 0.210 mmol) were added to DMF (3 mL) and the mixture was stirred at 25 ℃ for 1 h. o The reaction was stirred at 4°C for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 5 (26 mg, white solid) in a 22% yield.

[0185] MS m / z(ESI) = 570.4 [M+H] + .

[0186] 1H NMR (400 MHz, Chloroform- d ) δ 8.21 (d, J = 18.8 Hz, 1H), 6.85 (d, J = 3.2 Hz, 2H), 6.69 (dd, J = 18.8, 4.4 Hz, 2H), 5.48 (d, J = 1.6 Hz, 1H), 4.48 - 4.31 (m, 2H), 4.19 (dd, J = 11.2, 7.6 Hz, 1H), 4.14 - 3.86 (m, 7H), 3.76 (d, J = 3.2 Hz, 3H), 3.67 (dd, J = 9.6, 3.0 Hz, 1H), 3.16 - 3.02 (m,1H), 2.91 (d, J = 6.8 Hz, 2H), 2.25 (d, J = 2.4 Hz, 3H), 2.19 - 2.12 (m, 1H), 2.07 - 2.01 (m, 6H), 1.99 - 1.90 (m, 1H).

[0187] Example 6: Synthesis of Compound 6

[0188]

[0189] Synthesis of compound 6

[0190] Compound f (3.0 mg, 0.021 mmol), HATU (12.2 mg, 0.032 mmol), DIPEA (8.1 mg, 0.063 mmol) and compound 3-2 (10.0 mg, 0.021 mmol) were added to DMF (1 mL) and stirred at 25 ℃ for 1 h. o The reaction was stirred at 4°C for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 6 (2.2 mg, yellow solid) in a 17% yield.

[0191] MS m / z(ESI) = 600.4 [M+H] + .

[0192] 1 H NMR (400 MHz, Chloroform- d ) δ 9.73 (s, 1H),7.03 (d, J = 2.8 Hz,2H), 6.73 (s, 1H), 6.56 (s, 1H), 5.45 (s, 1H), 4.65 (d, J = 14.0 Hz, 1H),4.15 (q, J = 6.8 Hz, 4H), 4.01 (s, 3H), 3.70 (s, 8H), 3.00 (s, 2H), 2.81 (s,1H), 2.34 (s, 3H), 2.22 (d, J = 20.8 Hz, 6H), 1.48 (t, J = 6.8 Hz, 3H).

[0193] Example 7: Synthesis of Compound 7

[0194]

[0195] Synthesis of compound 7-1

[0196] Compound Int-A (198.1 mg, 0.47 mmol), compound a (354.7 mg, 1.42 mmol), cesium carbonate (462.7 mg, 1.42 mmol) and sodium iodide (212.8 mg, 1.42 mmol) were added to butanone (10 mL) and the mixture was stirred at 110 °C under nitrogen protection. o C. and stirred for 16 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by flash silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to obtain compound 7-1 (154 mg, yellow solid) in a yield of 55%.

[0197] MS m / z(ESI): 591.4[M+H] + .

[0198] Synthesis of compound 7-2

[0199] Compound 7-1 (154 mg, 0.26 mmol) was dissolved in dichloromethane (5 mL), followed by the addition of trifluoroacetic acid (1.5 mL) and the mixture was stirred at 25 oC for 1 hour. The reaction solution was concentrated under reduced pressure to obtain compound 7-2 (120 mg, yellow oil) with a yield of 94%.

[0200] MS m / z(ESI): 491.3[M+H] + .

[0201] Synthesis of compound 7

[0202] Compound 7-2 (44.2 mg, 0.09 mmol) was added to water (1 mL), and acetic acid (7.8 mg, 0.13 mmol) and potassium cyanate (34.9 mg, 0.43 mmol) were added. o C and stirred for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water solution containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 7 (12.4 mg, white solid) in a 26% yield.

[0203] MS m / z(ESI): 534.4[M+H] + .

[0204] 1 H NMR (400 MHz, Methanol- d 4) δ 6.96 (s, 1H), 6.94 - 6.87 (m, 2H), 6.72 (s, 1H), 5.59 (s, 1H), 4.63 - 4.54 (m, 1H), 4.23 - 4.12 (m, 4H), 3.85 (s, 3H), 3.75 (dd, J = 14.4, 4.0 Hz, 1H), 3.68 (s, 6H), 3.41 - 3.35 (m, 2H), 3.04 (t, J = 6.4 Hz, 2H), 2.28 (s, 4H), 2.25 (s, 4H), 1.43 (t, J = 7.2 Hz, 3H).

[0205] Example 8: Synthesis of Compound 8 and Compound 10

[0206]

[0207] Compound b (10.2 mg, 0.09 mmol), HATU (49.4 mg, 0.13 mmol), DIPEA (33.6 mg, 0.26 mmol) and compound 7-2 (44.2 mg, 0.09 mol) were added to DMF (2 mL) and the mixture was stirred at 25 ℃ for 1 h. o The reaction was stirred at 40 °C for 12 hours. After completion of the reaction, the reaction solution was separated and purified on a C18 reverse-phase column (acetonitrile / water solution containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 8 (10 mg, white solid, yield: 19%) and compound 10 (9 mg, white solid, yield: 17%), respectively.

[0208] Compound 8: MS m / z (ESI): 586.4 [M+H] + .

[0209] 1 H NMR (400 MHz, DMSO- d 6) δ 8.22 (t, J = 6.4 Hz, 1H), 7.62 (s, 1H), 7.05 (s, 1H), 6.89 (q, J = 2.8 Hz, 2H), 6.65 (s, 1H), 5.46 (s, 1H), 4.50 -4.40 (m, 1H), 4.16 - 4.00 (m, 4H), 3.78 (s, 3H), 3.72 - 3.65 (m, 2H), 3.61(s, 3H), 3.59 - 3.48 (m, 2H), 3.45 (d, J = 6.8 Hz, 1H), 2.98 (t, J = 5.6 Hz,2H), 2.68 (s, 1H), 2.20 (d, J = 18.4 Hz, 6H), 1.34 (t, J = 6.8 Hz, 3H).

[0210] Compound 10:

[0211] MS m / z(ESI): 586.3[M+H] + .

[0212] 1 H NMR (400 MHz, DMSO-d 6) δ 8.23 (t, J = 6.5 Hz, 1H), 7.63 (s, 1H), 7.05 (s, 1H), 6.90 (q, J = 2.8 Hz, 2H), 6.66 (s, 1H), 5.46 (s, 1H), 4.51 -4.40 (m, 1H), 4.16 - 4.00 (m, 4H), 3.79 (s, 3H), 3.72 - 3.66 (m, 2H), 3.61(s, 3H), 3.59 - 3.49 (m, 2H), 3.46 (d, J = 6.7 Hz, 1H), 2.99 (t, J = 5.5 Hz,2H), 2.69 (s, 1H), 2.22 (d, J = 18.3 Hz, 6H), 1.35 (t, J = 6.8 Hz, 3H).

[0213] Example 9: Synthesis of Compound 9

[0214]

[0215] Compound g (10.2 mg, 0.09 mmol), HATU (49.4 mg, 0.13 mmol), DIPEA (33.6 mg, 0.26 mmol) and compound 7-2 (44.2 mg, 0.09 mol) were added to DMF (2 mL) and the mixture was stirred at 25 ℃ for 1 h. o The reaction was stirred at 4°C for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 9 (22 mg, white solid) in a 42% yield.

[0216] MS m / z(ESI): 586.4[M+H] + .

[0217] 1 H NMR (400 MHz, Methanol- d 4) δ 7.94 (s, 1H), 6.95 (s, 1H), 6.89 (q, J= 3.2 Hz, 2H), 6.72 (s, 1H), 5.59 (s, 1H), 4.62 (dd, J = 14.0, 4.4 Hz, 1H),4.19 - 4.11 (m, 4H), 3.84 (s, 4H), 3.80 - 3.60 (m, 8H), 3.03 (t, J = 6.8 Hz,2H), 2.78 (d, J = 6.4 Hz, 1H), 2.28 (s, 3H), 2.25 (s, 3H), 1.42 (t, J = 7.2 Hz, 3H).

[0218] Example 10: Synthesis of Compound 11 and Compound 13

[0219]

[0220] Synthesis of compound 11-1

[0221] Compound Int-B (151 mg, 0.35 mmol), compound a (260 mg, 1.04 mmol), cesium carbonate (339 mg, 1.04 mmol) and sodium iodide (156 mg, 1.04 mmol) were added to butanone (10 mL) and stirred at 110 °C under nitrogen protection. o C. and stirred for 16 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by flash silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to obtain compound 11-1 (90 mg, yellow solid) in a yield of 43%.

[0222] MS m / z(ESI): 601.4[M+H] + .

[0223] Synthesis of compound 11-2

[0224] Compound 11-1 (90 mg, 0.15 mmol) was dissolved in dichloromethane (6 mL), and trifluoroacetic acid (2 mL) was added and the mixture was stirred at 25 °C. o C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain compound 11-2 (70 mg, yellow oil) with a yield of 93%.

[0225] MS m / z(ESI): 501.3[M+H] + .

[0226] Synthesis of Compound 11 and Compound 13

[0227] Compound b (5.0 mg, 0.044 mmol), HATU (25.1 mg, 0.066 mmol), DIPEA (16.8 mg, 0.130 mmol) and compound 11-2 (22.0 mg, 0.044 mmol) were added to DMF (2 mL) and stirred at 25 °C for 1 h. o The reaction was stirred at 40 °C for 12 hours. After completion of the reaction, the reaction solution was separated and purified via a C18 reverse-phase column (acetonitrile / water solution containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 11 (5 mg, yellow solid, yield: 19%) and compound 13 (7 mg, white solid, yield: 27%), respectively.

[0228] Compound 11:

[0229] MS m / z(ESI): 596.4[M+H] + .

[0230] 1 H NMR (400 MHz, Methanol- d 4) δ 8.14 (s, 1H), 6.99 (s, 2H), 6.88 (s,1H), 6.67 (s, 1H), 5.45 (s, 1H), 4.68 (t, J = 9.1 Hz, 1H), 4.45 - 4.29 (m,2H), 4.25 - 4.04 (m, 6H), 3.90 - 3.80 (m, 1H), 3.66 (s, 3H), 3.16 - 3.10 (m,1H), 2.93 (t, J = 6.6 Hz, 2H), 2.15 (s, 6H), 1.98 - 1.91 (m, 1H), 1.40 (t, J = 7.0 Hz, 3H), 1.06 - 0.96 (m, 2H), 0.79 - 0.70 (m, 2H).

[0231] Compound 13:

[0232] MS m / z(ESI): 596.4[M+H] + .

[0233] 1H NMR (400 MHz, Methanol- d 4) δ 8.15 (s, 1H), 7.00 (s, 2H), 6.88 (s,1H), 6.68 (s, 1H), 5.46 (s, 1H), 4.69 (t, J = 9.2 Hz, 1H), 4.45 - 4.29 (m,2H), 4.25 - 4.03 (m, 6H), 3.90 - 3.81 (m, 1H), 3.67 (s, 3H), 3.17 - 3.10 (m,1H), 2.93 (t, J = 6.4 Hz, 2H), 2.17 (s, 6H), 1.97 - 1.89 (m, 1H), 1.41 (t, J = 7.2 Hz, 3H), 1.06 - 0.95 (m, 2H), 0.79 - 0.69 (m, 2H).

[0234] Example 11: Synthesis of Compound 12

[0235]

[0236] Compound g (5.0 mg, 0.044 mmol), HATU (25.1 mg, 0.066 mmol), DIPEA (16.8 mg, 0.130 mmol) and compound 11-2 (22.0 mg, 0.044 mmol) were added to DMF (2 mL) and stirred at 25 °C for 1 h. o The reaction was stirred at 4°C for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water solution containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 12 (11 mg, white solid) in a 42% yield.

[0237] MS m / z(ESI): 596.4[M+H] + .

[0238] 1 H NMR (400 MHz, Methanol- d 4) δ 7.94 (s, 1H), 7.05 (d, J= 3.6 Hz,2H), 6.95 (s, 1H), 6.67 (s, 1H), 5.53 (s, 1H), 4.63 (dd, J = 14.0, 4.4 Hz,1H), 4.22 - 4.10 (m, 4H), 3.88 - 3.62 (m, 8H), 3.03 (t, J = 6.8 Hz, 2H), 2.82- 2.73 (s, 1H), 2.25 (d, J = 15.6 Hz, 6H), 1.99 - 1.89 (m, 1H), 1.42 (t, J =7.2 Hz, 3H), 1.10 - 1.00 (m, 2H), 0.79 - 0.71 (m, 2H).

[0239] Example 12: Synthesis of Compound 14

[0240]

[0241] Synthesis of compound 14

[0242] Compound d (2.5 mg, 0.02 mmol), HATU (11.4 mg, 0.03 mmol), DIPEA (7.8 mg, 0.06 mmol) and compound 3-2 (9.5 mg, 0.02 mmol) were added to DMF (1 mL) and stirred at 25 ℃ for 1 h. o The reaction was stirred at 4°C for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 14 (2.2 mg, white solid) in a 19% yield.

[0243] MS m / z(ESI): 584.4[M+H] + .

[0244] 1 H NMR (400 MHz, Chloroform- d ) δ 9.43 (s, 1H), 7.02 (d, J = 8.4 Hz,2H), 6.67 (d, J = 26.8 Hz, 2H), 5.47 (s, 1H), 4.39 (d,J = 14.0 Hz, 1H), 4.15(q, J = 6.8 Hz, 4H), 4.07 - 3.91 (m, 2H), 3.83 (d, J = 8.4 Hz, 1H), 3.73 (s,5H), 3.14 (s, 1H), 2.97 (d, J = 6.8 Hz, 2H), 2.44 (s, 3H), 2.33 (s, 3H), 2.30 (s, 3H), 2.13 (s, 3H), 1.48 (t, J = 7.2 Hz, 3H).

[0245] Example 13: Synthesis of Compound 15

[0246]

[0247] Synthesis of compound 15

[0248] Compound c (2.4 mg, 0.021 mmol), HATU (12.2 mg, 0.032 mmol), DIPEA (8.1 mg, 0.063 mmol) and compound 3-2 (10.0 mg, 0.021 mmol) were added to DMF (1 mL) and the mixture was stirred at 25 ℃ for 1 h. o The reaction was stirred at 4°C for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water solution containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 15 (1.8 mg, yellow solid) in a 15% yield.

[0249] MS m / z(ESI): 569.3[M+H] + .

[0250] 1 H NMR (400 MHz, DMSO- d 6) δ 7.75 (d, J = 2.4 Hz, 1H), 6.94 (s, 1H), 6.85 (s, 2H), 6.66 (s, 1H), 6.59 (d, J= 2.4 Hz, 1H), 5.32 (s, 1H), 4.68 -4.42 (m, 4H), 4.15 - 4.02 (m, 3H), 4.00 - 3.89 (m, 3H), 3.61 (s, 3H), 3.25 -3.22 (m, 1H), 2.89 (t, J = 6.4 Hz, 2H), 2.21 (s, 3H), 1.96 (d, J = 5.2 Hz,6H), 1.33 (t, J = 6.8 Hz, 3H).

[0251] Example 14: Synthesis of Compound 16

[0252]

[0253] Compound h (5.6 mg, 0.044 mmol), HATU (25.1 mg, 0.066 mmol), DIPEA (16.8 mg, 0.130 mmol) and compound 11-2 (22.0 mg, 0.044 mmol) were added to DMF (2 mL) and stirred at 25 ℃ for 1 h. o The reaction was stirred at 4°C for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water solution containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 16 (8 mg, white solid) in a 30% yield.

[0254] MS m / z(ESI): 610.3 [M+H] + .

[0255] 1 H NMR (400 MHz, Methanol- d 4) δ 7.97 (s, 1H), 7.12 (d, J = 3.3 Hz,2H), 6.98 (s, 1H), 6.69 (s, 1H), 5.55 (s, 1H), 4.66-4.61 (m, 1H), 4.20 - 4.10(m, 4H), 3.89 - 3.64 (m, 8H), 3.03 (t, J = 6.6 Hz, 2H), 2.80 - 2.73 (s, 1H), 2.55 (s, 3H), 2.28 (d,J = 15.2 Hz, 6H), 2.01 - 1.91 (m, 1H), 1.42 (t, J =7.0 Hz, 3H), 1.11 - 1.00 (m, 2H), 0.82 - 0.74 (m, 2H).

[0256] Example 15: Synthesis of Compound 17

[0257]

[0258] Synthesis of compound 17

[0259] Compound 1-2 (41.5 mg, 0.09 mmol) was added to water (1 mL), and acetic acid (7.8 mg, 0.13 mmol) and potassium cyanate (34.9 mg, 0.43 mmol) were added. o C and stirred for 12 hours. After completion of the reaction, the reaction solution was purified via a C18 reverse-phase column (acetonitrile / water solution containing 10 mM ammonium bicarbonate = 20%-50%; purification method: RP4-25cm; flow rate: 40 mL / min; gradient time: 15 minutes) to obtain compound 17 (16.8 mg, white solid) in a 37% yield.

[0260] MS m / z(ESI): 504.4[M+H] + .

[0261] 1 H NMR (400 MHz, Methanol- d 4) δ 7.02 (s, 1H), 6.98 - 6.92 (m, 2H), 6.76 (s, 1H), 5.59 (s, 1H), 4.63 - 4.54 (m, 1H), 4.23 - 4.12 (m, 4H), 3.85(s, 3H), 3.78-3.72 (m, 1H), 3.68 (s, 6H), 3.41 - 3.35 (m, 2H), 3.04 (s, 3H), 2.28 (s, 4H), 2.25 (s, 4H).

[0262] Comparative Example 1: Synthesis of RPL554

[0263]

[0264] RPL554 was synthesized according to the preparation method provided in invention patent WO 00 / 58308A1.

[0265] MS m / z(ESI): 478.2[M+H] + .

[0266] 1 H NMR (400 MHz, CDCl 3 ) δ 6.89 (s, 2H), 6.70 (s, 1H), 6.67 (s, 1H), 5.45 (s, 1H), 4.41 (t, J = 6.99 Hz, 2H), 4.07-4.02 (m, 2H), 3.90 (s, 3H), 3.76 (s, 3H), 3.58-3.49 (m, 2H), 2.91 (t, J = 6.17 Hz, 2H), 2.28 (s, 3H), 2.06 (s, 6H).

[0267] Experimental Example 1: Enzyme Inhibitory Activity Test of Representative Compounds against PDE3A and PDE4B

[0268] 1) Materials and Equipment:

[0269]

[0270] 2) Experimental methods:

[0271] (1) Compound dilution: First, prepare a 10 μM compound stock solution with DMSO. When testing the single-concentration inhibition rate, directly administer the compound according to the corresponding dilution ratio. When testing the enzyme activity IC 50 The compound concentrated stock solution was diluted from 10 μM in a 4-fold concentration gradient to obtain 10 concentration points.

[0272] (2) Prepare the incubation system according to the table below, mix well, and incubate at room temperature for 1 h.

[0273]

[0274] (3) After incubation, 100 μL of Binding Agent was added to each group to dilute the above system, mixed well, and incubated at room temperature for another 1 h.

[0275] (4) After the second round of incubation, the sample was placed in a microplate reader and the fluorescence signal was detected according to the test conditions of excitation wavelength 490 nm and emission wavelength 520 nm. The fluorescence signal was calculated according to the %Inhibition rate = (FP B −FP S ) / (FP B −FPV ) × 100% to calculate the inhibition rate.

[0276] 3) Test results

[0277]

[0278] It has been verified that the compound provided by the present invention has good inhibitory activity against both PDE3A and PDE4B, and is superior to RPL554.

[0279] Experimental Example 2: Agonist activity of representative compounds in increasing intracellular cAMP concentration

[0280] 1) Materials and Equipment:

[0281]

[0282] 2) Experimental methods:

[0283] (1) Add the compound 30 minutes before adding Forskolin, then add the remaining components except the compound in the table, mix gently, and incubate at room temperature for another 1 hour.

[0284] The incubation system is as follows:

[0285]

[0286] (2) After the incubation, the fluorescence signal was detected using a microplate reader under the test conditions of excitation wavelength of 340 nm and emission wavelengths of 616 nm and 665 nm, and the data was read.

[0287] 3) Test results

[0288]

[0289] It has been verified that the compound provided by the present invention has better and higher cAMP agonist activity.

[0290] Experimental Example 3: Inhibitory effect of representative compounds on the secretion of inflammatory factor TNF-α

[0291] 1) Materials and Equipment:

[0292]

[0293] 2) Experimental methods:

[0294] (1) Mononuclear cells (concentration of 10 6 / ml) were suspended in RPMI 1640 medium containing 1% fetal bovine serum and seeded into 96-well plates (10 5 The cells were incubated at 37°C with 5% CO2 for 2 hours to allow the cells to adhere.

[0295] (2) Adherent cells were cultured in fresh RPMI 1640 medium containing 1% fetal bovine serum at 5% CO2 and 37°C, containing LPS (10 ng / ml) and compounds (maximum final concentration 10 μM, 4-fold dilution).

[0296] (2) After 18 hours, collect the cell supernatant. According to the instructions of the TNF-α enzyme-linked immunosorbent assay (ELISA) kit, detect the TNF-α concentration in the supernatant and calculate the IC 50 .

[0297] 3) Test results

[0298]

[0299] It has been verified that the compound provided by the present invention has a good effect of inhibiting the release of TNF-α.

Claims

1. A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof: in, R1 is selected from -NR4R5 or cyclic Cy; R4 and R5 are independently selected from H, C1-C6 alkyl; Ring Cy is selected from 4-7 membered heteroaryl groups containing 1-5 heteroatoms selected from N, wherein the 4-7 membered heteroaryl groups containing 1-5 heteroatoms selected from N are optionally substituted by 0, 1, 2, 3, 4 or 5 R 1a replace; Each R 1a are the same or different and are each independently selected from H, C1-C6 alkyl, and hydroxyl; R2 is selected from H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy; R3 is selected from H, C1-C6 alkyl; X is -(CH2) m -; m is 1; Y is -(CH2) n -; n is 1 or 2.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The compound has a structure shown in formula (II) or formula (III): 。 3. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein Said R4 and R5 are independently selected from H, C1-C4 alkyl; said ring Cy is selected from 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, said 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N is optionally substituted by 0, 1, 2 or 3 R 1a replace.

4. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein The ring Cy is selected from Where w is 0, 1 or 2, v is 0, 1, 2, 3, 4 or 5, and u is 0, 1, 2 or 3; R 1a Each is independently selected from H, C1-C4 alkyl and hydroxyl.

5. The compound according to claim 4 or a pharmaceutically acceptable salt thereof, wherein The ring Cy is selected from:

6. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R2 is selected from H, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy.

7. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R3 is selected from H, C1-C4 alkyl.

8. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein In formula (I), formula (II) or formula (III), R4 and R5 are independently selected from H, C1-C4 alkyl; ring Cy is selected from 5-6 membered heteroaryl containing 1-3 N heteroatoms, and the 5-6 membered heteroaryl containing 1-3 N heteroatoms is optionally replaced by 0, 1 or 2 R 1a replace; R 1a Each independently selected from H, C1-C4 alkyl, hydroxyl; R2 is selected from H, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, R3 is selected from H, C1-C4 alkyl.

9. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein In formula (I), formula (II) or formula (III), R4 and R5 are independently selected from H, C1-C4 alkyl; ring Cy is selected from Where w is 0, 1 or 2, v is 0, 1, 2, 3, 4 or 5, and u is 0, 1, 2 or 3; R 1a Each independently selected from H, C1-C4 alkyl, hydroxyl; R2 is selected from H, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy; R3 is selected from H, C1-C4 alkyl.

10. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein In formula (I), formula (II) or formula (III), R4 and R5 are independently selected from H, C1-C4 alkyl; ring Cy is selected from Where w is 0, 1 or 2, v is 0, 1, 2, 3, 4 or 5, and u is 0, 1, 2 or 3; R 1a Each independently selected from H, C1-C4 alkyl and hydroxyl; R2 is selected from C1-C3 alkyl, C3-C4 cycloalkyl, C1-C3 alkoxy; R3 is selected from C1-C3 alkyl.

11. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein In formula (I), formula (II) or formula (III), R4 and R5 are independently selected from H, C1-C4 alkyl; and ring Cy is selected from: R2 is selected from H, C1-C4 alkyl, C3-C5 cycloalkyl, C1-C4 alkoxy, R3 is selected from H, C1-C4 alkyl.

12. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein In formula (I), formula (II) or formula (III), R4 and R5 are independently selected from H, C1-C4 alkyl; and ring Cy is selected from: R2 is selected from C1-C3 alkyl, C3-C4 cycloalkyl, C1-C3 alkoxy; R3 is selected from C1-C3 alkyl.

13. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The compound represented by general formula (I) is selected from: 14 . A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient, carrier or diluent.

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