Barbituric acid derivative and preparation method thereof

Abstract

The invention provides a barbituric acid derivative as shown in a formula I, and a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug of the barbituric acid derivative. The derivative can inhibit the activity of cardiac myosin ATP enzyme and relieve myocardial hypertrophy caused by excessive contraction of myocardial force, and is used for treating hypertrophic cardiomyopathy or heart diseases with one or more pathophysiology characteristics related to the hypertrophic cardiomyopathy. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to a myosin inhibitor of a barbituric acid derivative, a preparation method and application thereof. Background Art

[0002] Hypertrophic cardiomyopathy (HCM) is a myocardial disease of unknown cause, characterized by asymmetric hypertrophy of the ventricular wall, which often invades the ventricular septum, shrinks the ventricular inner cavity, obstructs left ventricular blood filling, and decreases left ventricular diastolic compliance. Depending on whether there is obstruction in the left ventricular outflow tract, it is divided into obstructive and non-obstructive hypertrophic cardiomyopathy, which may be related to genetics. Hypertrophic cardiomyopathy carries a risk of sudden death and is one of the causes of sudden death during exercise. In addition, HCM is also one of the most common heart diseases in cats, with a higher incidence in cats such as American Shorthair, Persian, and Maine Coons, which can lead to sudden death in cats.

[0003] The cause of hypertrophic cardiomyopathy is still unclear. It is currently believed that genetic factors are the main cause. The basis is that the disease has a clear familial tendency to occur, and is often combined with other congenital cardiovascular malformations. Some patients have the disease at birth, and the genetic genotype of HLA antigens can be seen in patients with the disease. Familial cases are transmitted in the form of autosomal dominant inheritance. The type of onset can be asymptomatic asymmetric myocardial hypertrophy, or there can be typical obstructive symptoms. About 50% to 55% of patients with hypertrophic cardiomyopathy have a family history, which is an autosomal dominant genetic disease. The genetic theory of hypertrophic cardiomyopathy has been recognized and is believed to be closely related to the tissue compatibility antigen system. The detection rate of HLA-DRW4, Ag, and B5 antigens in patients with hypertrophic cardiomyopathy has increased significantly. In 1995, the World Health Organization and the International Heart Association's Heart Nomenclature and Classification Special Committee clearly pointed out that it was the gene mutation of the sarcomere contractile protein that caused familial hypertrophic cardiomyopathy.

[0004] HCM-related gene mutations can cause myocardial remodeling, myocardial fiber disorder and abnormal morphology, and compensatory hyperplasia and hypertrophy of myocardial cells by changing myocardial contractility, affecting myocardial cell sensitivity to calcium ions, and affecting myocardial cell energy metabolism.

[0005] Myosin and actin are the material basis of myocardial contraction. Myosin cross bridges periodically bind and dissociate with actin, driving myofilaments to slide and leading to myocardial contraction. Myosin has ATPase activity and provides power for myocardial contraction by hydrolyzing ATP. Myosin mutations can lead to prolonged binding time of myosin and actin, excessive contraction and impaired relaxation of the left ventricular myocardium, leading to left ventricular hypertrophy and fibrosis, and inducing HCM. Inhibition of myosin ATPase can produce a negative inotropic effect and alleviate pathological changes such as myocardial hypertrophy caused by excessive contraction of the left ventricular myocardium.

[0006] At present, the treatment methods for HCM mainly include drugs and non-drug treatments, which can reduce myocardial contractility, alleviate outflow tract stenosis, and improve cardiac diastolic function. Non-drug treatments mainly include ventricular septal ablation and ventricular septal hypertrophy resection. Drug treatments include beta-blockers, calcium channel blockers, disopyramide, etc. The above drugs are mainly used to improve symptoms, but cannot target the cause, delay the progression of myocardial hypertrophy, or change the prognosis, and the treatment effect is limited. There is still a lack of ideal treatment methods for HCM in the clinic.

[0007] MYK-461 is a new type of myosin allosteric inhibitor that can inhibit myocardial overcontraction caused by HCM and relieve left ventricular outflow tract obstruction. However, it is eliminated slowly from the body and the drug stays in the body for too long, making it difficult to quickly adjust the dose. Therefore, the development of new myosin inhibitors with better activity and pharmacokinetics has important clinical value and significance. Summary of the invention

[0008] The object of the present invention is to provide a novel compound for use as an inhibitor of cardiac myosin ATPase for treating hypertrophic cardiomyopathy or a heart disease having one or more pathophysiological features associated with hypertrophic cardiomyopathy.

[0009] In the first aspect of the present invention, there is provided a compound of formula I, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof:

[0010]

[0011] in,

[0012] X 1 and X 2 are ring atoms and are each independently selected from N or C;

[0013] Group fragment In this case, A does not exist, and X 1 and X 2 Each is independently R 5 and R 6 or when A exists, A and X 1 and X 2 The ring atoms together form a 3-10 membered heterocycloalkyl, a 6-10 membered aryl or a 5-10 membered heteroaryl, wherein the heteroatoms are independently selected from at least one of N, O and S; and the A is further replaced by one or more R a Replace, when R a When it is multiple, the R a Same or different;

[0014] R a , R 5 and R6 Each independently selected from H, halogen, hydroxy, amino, cyano, oxo, C 1 -C 6 Alkyl, C 3 -C 8 Cycloalkyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl, C 3 -C 8 Halogenated cycloalkyl and C 1 -C 6 Haloalkoxy; the C 1 -C 6 Alkyl, C 3 -C 8 Cycloalkyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl, C 3 -C 8 Halogenated cycloalkyl and C 1 -C 6 The haloalkoxy group is optionally substituted with one or more R b Substitution; said R b A substituent selected from the group consisting of: H, halogen, hydroxy, amino, nitro, cyano, oxo, carboxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl, C 1 -C 6 Haloalkoxy; when R b When it is multiple, the R b Same or different;

[0015] R 1 Selected from H, halogen, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 3 -C 8 Cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aryl and 5-10 membered heteroaryl, the C 1 -C 6 Alkyl, C2 -C 6 Alkenyl, C 3 -C 8 Cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aryl and 5-10 membered heteroaryl are optionally substituted by one or more R c Substitution; said R c A substituent selected from the group consisting of: H, halogen, hydroxy, amino, nitro, cyano, oxo, carboxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl, C 1 -C 6 Haloalkoxy; when R c When it is multiple, the R c Same or different;

[0016] R 2 Selected from H and C 1 -C 6 alkyl;

[0017] R 3 is selected from H, halogen, hydroxy, amino, nitro, cyano, oxo, C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl and C 1 -C 6 Haloalkoxy, the C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl and C 1 -C 6 The haloalkoxy group is optionally substituted with one or more R d Substitution; said R d A substituent selected from the group consisting of: H, halogen, hydroxy, amino, nitro, cyano, oxo, carboxyl, C 1 -C 6 Alkyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, C 1 -C6 Haloalkyl, C 1 -C 6 Haloalkoxy; when R d When it is multiple, the R d Same or different;

[0018] m and n are 1, 2 or 3 respectively.

[0019] In the second aspect of the present invention, there is provided a compound of formula II, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof:

[0020]

[0021] Among them, X 1 , X 2 , A, R 1 , R 2 , R 3 and n are as defined in the first aspect of the present invention.

[0022] In the present invention, the definitions of certain substituents in the compound as shown in Formula I can be as described below, and the definitions of substituents not mentioned are as described in any of the above schemes.

[0023] In a preferred embodiment of the present invention, the barbituric acid derivative has a structure as shown in the following formula I-1:

[0024] Among them, R 5 and R 6 Each independently selected from H, halogen, hydroxy, amino, cyano, oxo, C 1 -C 6 Alkyl, C 3 -C 8 Cycloalkyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl, C 3 -C 8 Halogenated cycloalkyl and C 1 -C 6 Haloalkoxy; the C 1 -C 6 Alkyl, C 3 -C 8 Cycloalkyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, C 1 -C6 Haloalkyl, C 3 -C 8 Halogenated cycloalkyl and C 1 -C 6 The haloalkoxy group is optionally substituted with one or more R b Replacement; R 1 , R 2 , R 3 , R b , m and n are defined as described in the first aspect of the present invention.

[0025] In a preferred embodiment of the present invention, the barbituric acid derivative has a structure as shown in the following formula I-2:

[0026] Among them, A and X 1 and X 2 The ring atoms together form a 3- to 10-membered heterocycloalkyl, a 6- to 10-membered aryl or a 5- to 10-membered heteroaryl, wherein the heterocyclic ring is independently selected from at least one of N, O and S; and the A is further replaced by one or more R a Replace, when R a When it is multiple, the R a Same or different; R 1 , R 2 , R 3 , R a , m and n are defined as described in the first aspect of the present invention.

[0027] In a preferred embodiment of the present invention, the group fragment In the above example, A exists and A and X 1 and X 2 The ring atoms together form a 3- to 8-membered heterocycloalkyl group, a 6- to 8-membered aryl group, or a 5- to 8-membered heteroaryl group.

[0028] In a preferred embodiment of the present invention, the 3- to 8-membered heterocycloalkyl, 6- to 8-membered aryl or 5- to 8-membered heteroaryl has 1 to 3 heteroatoms selected from N, O and S.

[0029] Preferably, the group fragment In the 1 and X 2 Ring atoms together form

[0030] In a preferred embodiment of the present invention, R 1 Selected from C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 3 -C 8Cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aryl and 5-10 membered heteroaryl, the C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 3 -C 8 Cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aryl and 5-10 membered heteroaryl are optionally substituted by 1, 2 or 3 R c Substitution; said R c A substituent selected from the group consisting of: H, halogen, hydroxy, amino, nitro, cyano, oxo, carboxyl, C 1 -C 3 Alkyl, C 2 -C 4 Alkenyl, C 1 -C 3 Alkoxy, C 1 -C 3 Haloalkyl, C 1 -C 3 Haloalkoxy; when R c When it is multiple, the R c Same or different.

[0031] In a preferred embodiment of the present invention, R 1 Selected from C 1 -C 3 Alkyl, C 2 -C 4 Alkenyl, C 3 -C 6 Cycloalkyl, 3- to 8-membered heterocycloalkyl, 5- to 8-membered aryl and 5- to 8-membered heteroaryl, the C 1 -C 3 Alkyl, C 2 -C 4 Alkenyl, C 3 -C 6 Cycloalkyl, 3- to 8-membered heterocycloalkyl, 5- to 8-membered aryl and 5- to 8-membered heteroaryl are optionally substituted by one R c Substitution; said R c A substituent selected from the group consisting of: H, halogen, hydroxy, amino, nitro, cyano, oxo, carboxyl, C 1 -C 3 Alkyl, C 2 -C 4 Alkenyl, C 1 -C 3 Alkoxy, C 1 -C 3 Haloalkyl, C 1 -C 3 Halogenated alkoxy.

[0032] In a preferred embodiment of the present invention, R 1 Selected from C 1 -C 6 Alkyl, C 3 -C 6 Cycloalkyl, C 1 -C 6 Alkyl substituted C 3 -C 6 Cycloalkyl, 4- to 6-membered heterocycloalkyl, 5- to 6-membered aryl, 5- to 6-membered heteroaryl, 1 -C 6 Alkyl-substituted 5- to 6-membered aromatic group, C 1 -C 6 Alkyl-substituted 5- to 6-membered heteroaryl;

[0033] In a preferred embodiment of the present invention, R 1 Selected from

[0034] In a preferred embodiment of the present invention, R 2 Selected from methyl.

[0035] In a preferred embodiment of the present invention, R 3 is selected from the group consisting of H, F, Cl, Br, hydroxy, amino, nitro, cyano, oxo, methyl, methoxy, trifluoromethyl and trifluoromethoxy.

[0036] In a preferred embodiment of the present invention, the barbituric acid derivative is selected from any one of the following compounds:

[0037]

[0038]

[0039] In a preferred embodiment of the present invention, the barbituric acid derivative is selected from any one of the following compounds:

[0040]

[0041]

[0042] In the third aspect of the present invention, there is provided a method for preparing the barbituric acid derivatives represented by formula II of the second aspect of the present invention, their tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, comprising the steps of:

[0043]

[0044] Among them, X 1 , X 2 , A, R 1, R 2 , R 3 and n are defined as described in the second aspect of the present invention.

[0045] In the fourth aspect of the present invention, a pharmaceutical composition is provided, comprising: the barbituric acid derivatives as described in the first aspect or the second aspect of the present invention, their tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs; and a pharmaceutically acceptable carrier.

[0046] In the fifth aspect of the present invention, there is provided the use of the barbituric acid derivatives, their tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs as described in the first aspect or the second aspect of the present invention in the preparation of drugs, or the use of the pharmaceutical composition as described in the fourth aspect of the present invention in the preparation of drugs, the uses including: for the treatment of cardiac diastolic and / or systolic dysfunction, more preferably for the treatment of heart diseases associated with left ventricular hypertrophy and / or diastolic dysfunction, and further preferably for the treatment of hypertrophic cardiomyopathy.

[0047] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention.

[0048] Terms and Definitions

[0049] Unless otherwise specified, the definitions of groups and terms recorded in the specification and claims of this application, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., can be arbitrarily combined and combined with each other. The group definitions and compound structures after such combination and combination shall fall within the scope recorded in the specification of this application.

[0050] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those skilled in the art to which the subject matter of the claims pertains. Unless otherwise indicated, all patents, patent applications, and publications cited herein are incorporated herein by reference in their entirety. If there are multiple definitions of a term herein, the definition in this chapter shall prevail.

[0051] It should be understood that the above brief description and the detailed description below are exemplary and are only used for explanation, and do not impose any restrictions on the subject matter of the present invention. In this application, unless otherwise specifically stated, the use of the singular also includes the plural. It must be noted that unless otherwise clearly stated in the text, the singular form used in this specification and claims includes the plural form of the referred thing. It should also be noted that unless otherwise stated, the "or" and "or" used mean "and / or". In addition, the term "including" and other forms, such as "including", "containing" and "containing" are not restrictive.

[0052] Unless otherwise indicated, conventional methods within the technical scope of the art are adopted, such as mass spectrometry, NMR, IR and UV / VIS spectroscopy and pharmacological methods. Unless specifically defined, the terms used herein in the relevant descriptions of analytical chemistry, organic synthetic chemistry, and drugs and medicinal chemistry are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, preparation and delivery, and in the treatment of patients. For example, the instructions for use of the kit can be utilized by the manufacturer, or the reaction and purification can be implemented according to a manner well known in the art or the description of the present invention. Generally, the above-mentioned techniques and methods can be implemented according to conventional methods well known in the art according to the description in a plurality of summaries and more specific documents cited and discussed in this specification. In this specification, groups and substituents thereof can be selected by those skilled in the art to provide stable structural parts and compounds.

[0053] When substituents are described by conventional chemical formulas written from left to right, the substituents also include chemically equivalent substituents that would result if the formula were written from right to left. 2 O is equivalent to OCH 2 As used herein, or Represents the attachment site of a group. As used herein, "R 1 ”, “R1” and “R 1 " have the same meaning and can be used interchangeably. 2 Other symbols and similar definitions have the same meaning.

[0054] The section headings used herein are only for the purpose of organizing the article and should not be interpreted as limitations on the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals and papers, are incorporated herein by reference in their entirety.

[0055] In addition to the foregoing, when used in the specification and claims of the present application, the following terms have the meanings indicated below unless otherwise specifically stated.

[0056] In the present application, "optional" or "optionally" means that the event or situation described later may or may not occur, and the description includes both the occurrence and non-occurrence of the event or situation. For example, "optionally substituted aryl" means that aryl is substituted or unsubstituted, and the description includes both substituted aryl and unsubstituted aryl.

[0057] In the present application, the term "salt" or "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are suitable for use in contact with human and animal tissues within the scope of sound medical judgment without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0058] The term "tautomer" refers to functional group isomers resulting from the rapid movement of an atom in a molecule between two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Prototropic tautomers arise from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to separate a single tautomer usually produce a mixture whose physicochemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical characteristics within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; while in phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.

[0059] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, diastereomers and conformational isomers.

[0060] Depending on the choice of starting materials and methods, the compounds of the invention may exist in the form of one of the possible isomers or a mixture thereof, for example as a pure optical isomer, or as a mixture of isomers, such as a racemic and diastereomeric mixture, depending on the number of asymmetric carbon atoms. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to the chiral center (or multiple chiral centers) in the molecule. The prefixes D and L or (+) and (–) are the symbols used to specify the rotation of plane polarized light caused by the compound, where (–) or L indicates that the compound is levorotatory. Compounds prefixed with (+) or D are dextrorotatory.

[0061] When the bonds to chiral carbon in the formula of the present invention are depicted as straight lines, it should be understood that both the (R) and (S) configurations of the chiral carbon and the enantiomerically pure compounds and mixtures thereof produced therefrom are included within the scope of the general formula. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond. and straight dashed key Indicates the relative configuration of a stereocenter.

[0062] The term "solvate" means that the compound of the present invention or a salt thereof includes a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent forces between molecules. When the solvent is water, it is a hydrate.

[0063] The term "prodrug" refers to a compound of the present invention that can be converted into a biologically active compound under physiological conditions or by solvolysis. The prodrug of the present invention is prepared by modifying the functional groups in the compound, and the modification can be removed by conventional operations or in vivo to obtain the parent compound. The prodrug includes a compound formed by connecting a hydroxyl or amino group in the compound of the present invention to any group. When the prodrug of the compound of the present invention is administered to a mammalian subject, the prodrug is cleaved to form a free hydroxyl group and a free amino group, respectively.

[0064] The numerical ranges recorded in the specification and claims of this application, when the numerical range is understood as an "integer", should be understood as recording the two endpoints of the range and each integer in the range. For example, "an integer from 1 to 6" should be understood as recording each integer of 0, 1, 2, 3, 4, 5 and 6. When the numerical range is understood as a "number", it should be understood as recording the two endpoints of the range and each integer in the range and each decimal in the range. For example, "a number from 1 to 10" should be understood as recording not only each integer of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, but also at least recording the sum of each integer therein and 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, respectively.

[0065] The term "alkyl" when used alone or as part of another substituent means a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, having, for example, 1 to 6 carbon atoms, and connected to the rest of the molecule by a single bond. 1 -C 6 The term "alkyl" is used to refer to a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl and hexyl. The term "C 1 -C 3 "Alkyl" is used to represent a straight or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C1-3 alkyl group includes C 1 -C 2 Alkyl and C 2 -C 3 Alkyl, etc.; it may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1 -C 3 Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, and the like.

[0066] When used alone or as part of another substituent, the term "C 1 -C 6 "Haloalkyl" refers to an alkyl group as described above wherein any number (at least one) of the hydrogen atoms attached to the alkyl chain are replaced by fluorine, chlorine, bromine or iodine.

[0067] The term "cycloalkyl" when used alone or as part of another substituent refers to a cyclic alkyl group. m-n "Cycloalkyl" is understood to mean a saturated, unsaturated or partially saturated carbon ring having m to n atoms. For example, "3-8 membered cycloalkyl" or "C 3 -C 8 "Cycloalkyl" includes C 3-6 , C 3-5 , C 4-8 , C 4-6 , C 4-5 , C 5-8 or C 5-6 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. 3-8 Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [2.2.2]bicyclooctane, etc. The term "cycloalkyl" may be used interchangeably with the term "carbocyclyl".

[0068] The term "heterocycloalkyl" when used alone or as part of another substituent refers to a cycloalkyl group in which one or more (in some embodiments, 1 to 3) carbon atoms are replaced by heteroatoms such as, but not limited to, N, O, S, and P. The term "mn-membered heterocycloalkyl" or "C m-n "Heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having m to n atoms, wherein the heteroatoms are selected from N, O, S, P, preferably from N, O or S. For example, the term "3-8 membered heterocycloalkyl" or "C 3 -C 8 "Heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having 3 to 8 atoms, wherein 1, 2 or 3 ring atoms are selected from N, O, S, P, preferably from N, O or S. The term "heterocycloalkyl" can be used interchangeably with the term "heteroalkane ring".

[0069] The term "alkenyl" when used alone or as part of another substituent refers to a straight or branched chain monovalent hydrocarbon radical (e.g., C 2 -C 4) of two to forty carbon atoms having at least one carbon-carbon sp2 double bond. 2 -C 6 Alkenyl, such as C 2 -C 4Examples of alkenyl groups include, but are not limited to, vinyl and allyl.

[0070] The term "alkynyl" when used alone or as part of another substituent refers to a straight or branched chain monovalent hydrocarbon radical (e.g., C 2 to C 4) of two to forty carbon atoms having at least one carbon-carbon sp triple bond. 2 -C 6 Alkynyl, such as C 2 -C 4 Examples of alkynyl groups include, but are not limited to, ethynyl and propynyl.

[0071] The term "alkoxy" by itself or as part of another substituent refers to the group -OR Q , where R Q is "alkyl" as defined above.

[0072] As used herein, the term "halogen" alone or as part of another substituent refers to fluorine, chlorine, bromine, iodine; preferably fluorine or chlorine.

[0073] As used herein, the term "cyano" by itself or as part of another substituent means -CN.

[0074] As used herein, the term "amino" by itself or as part of another substituent means -NH 2 .

[0075] As used herein, the term "hydroxy" by itself or as part of another substituent refers to -OH.

[0076] The term "oxo" when used alone or as part of another substituent refers to the replacement of two hydrogen atoms on a methylene group by oxygen atoms, ie, the methylene group is replaced by a carbonyl group, representing =0.

[0077] The term "aryl" when used alone or as part of another substituent means a monocyclic, bicyclic and tricyclic carbocyclic ring system containing 6-14 ring atoms, or 6-10 ring atoms. Examples of aryl groups can include phenyl, naphthyl and anthracenyl. Unless otherwise specified, the group "6-10 membered aryl" means an aryl group containing 6-10 ring carbon atoms. The term "aryl" can be used interchangeably with the term "aromatic ring".

[0078] The term "heteroaryl" when used alone or as part of other substituents refers to monocyclic, bicyclic and tricyclic ring systems containing 5-6 ring atoms, or 5-10 ring atoms, or 5-12 ring atoms, wherein at least one ring contains one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, the heteroaryl group can be attached to the rest of the molecule (e.g., the main structure in the general formula) through any reasonable position (which can be C in CH, or N in NH). Examples include, but are not limited to, furanyl, imidazolyl, isoxazolyl, oxazolyl, pyrrolyl, pyrazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, thienyl, thiazolyl, etc.; also include the following bicyclic rings, but are by no means limited to these bicyclic rings: benzimidazolyl, benzofuranyl, benzothienyl, indolyl, oxoindolyl, imidazopyridinyl, pyrazolopyridinyl, pyrazolopyrimidinyl, quinolyl, isoquinolyl, quinazolinyl, etc. The term "heteroaryl" may be used interchangeably with the terms "heteroaromatic ring", "heteroaromatic ring" or "heteroaromatic ring group".

[0079] In this application, "pharmaceutical composition" refers to a preparation of the compound of the present invention and a medium generally accepted in the art for delivering the biologically active compound to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to promote administration of the organism, facilitate the absorption of the active ingredient, and thus exert biological activity.

[0080] In this application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the relevant governmental regulatory authorities as acceptable for human or livestock use.

[0081] The term "excipient" refers to a pharmaceutically acceptable inert ingredient. Examples of the term "excipient" include, but are not limited to, binders, disintegrants, lubricants, glidants, stabilizers, fillers, and diluents. Excipients can enhance the handling characteristics of a pharmaceutical formulation, i.e., make the formulation more suitable for direct compression by increasing fluidity and / or adhesion.

[0082] The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) ameliorating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade leading to or causing the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0083] The term "prevent" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0084] The term "patient" refers to any animal, preferably a mammal, that is about to or has been administered the compound or composition according to embodiments of the present invention. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc.

[0085] The term "therapeutically effective amount" refers to an amount of a compound that is effective in treating a disease or condition described herein when administered to a patient."Therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, and can be adjusted as needed by those skilled in the art.

[0086] The reaction temperature of each step can be appropriately selected according to the solvent, starting materials, reagents, etc., and the reaction time can also be appropriately selected according to the reaction temperature, solvent, starting materials, reagents, etc. After the reaction of each step is completed, the target compound can be separated and purified from the reaction system by common methods, such as filtration, extraction, recrystallization, washing, silica gel column chromatography, etc. In the case of not affecting the next step reaction, the target compound can also be directly used in the next step reaction without separation and purification.

[0087] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0088] Beneficial Effects

[0089] After extensive and in-depth research, the inventors unexpectedly developed a barbituric acid derivative. It is a small molecule inhibitor of cardiac myosin ATPase, which can reduce myocardial hypertrophy caused by excessive myocardial contraction by inhibiting the activity of cardiac myosin ATPase. The main mechanism of action is to reduce the ATPase activity of β-cardiac myosin by inhibiting the release of Pi and stabilizing the myosin-ADP-Pi intermediate. At the same time, it can also regulate other steps of the myosin ATPase cycle, reduce the rate-limiting step of the ATP cycle (the release of Pi), and thus reduce the activity of the ATPase cycle. The barbituric acid derivatives provided in the present application also reduce the rate of myosin releasing ADP and the binding rate with actin in the ADP-bound state.

[0090] The compound of formula I provided by the present invention, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug can be used to treat cardiac diastolic and / or systolic dysfunction, more preferably for treating heart diseases associated with left ventricular hypertrophy and / or diastolic dysfunction, and further preferably for treating hypertrophic cardiomyopathy. DETAILED DESCRIPTION

[0091] The present invention is further described below in conjunction with specific examples. It should be understood that the following description is only the most preferred embodiment of the present invention and should not be considered as limiting the scope of protection of the present invention. On the basis of a full understanding of the present invention, the experimental methods in the following examples that do not specify specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer. Those skilled in the art may make non-essential changes to the technical solution of the present invention, and such changes should be deemed to be included in the scope of protection of the present invention.

[0092] Intermediate A1: Preparation of 1-benzyl-6-chloro-3-ethylpyrimidine-2,4(1H,3H)-dione

[0093] The synthetic route of intermediate A1 is as follows:

[0094]

[0095] Step 1: Preparation of intermediate A1-3

[0096] Add acetic acid (270 mL) to a three-necked flask, then add malonic acid (120 g) and N-ethylurea (89 g), heat to 70°C, add acetic anhydride (195 mL) dropwise, stir, continue heating to 90°C, stir for 3 hours until the reaction is complete, cool to 50°C, concentrate the reaction solution under reduced pressure, then add ethanol (180 mL), cool the mixture to 0°C, stir for 2 hours. Filter and dry to obtain intermediate A1-3 (123.3 g, yield 78%).

[0097] ESI-MS (m / z): 156.2.

[0098] 1 H-NMR (400MHz, DMSO-d6, δ / ppm): 11.30 (s, 1H), 3.70 (q, 2H), 3.59 (s, 2H), 1.06 (t, 3H).

[0099] Step 2: Preparation of intermediate A1-4

[0100] Add POCl to the intermediate A1-3 (62.3 g) 3 (380 mL), water (10.8 mL) was added dropwise to the suspension at 0°C, and then the reaction mixture was heated at 80°C for 5 hours. The resulting brown solution was slowly cooled to room temperature, and POCl 3 The residue was washed with methanol (200 mL) and the obtained solid was recrystallized from ethanol to give Intermediate A1-4 (50.2 g, yield 72%).

[0101] ESI-MS (m / z): 174.7.

[0102] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 12.34 (s, 1H), 5.89 (s, 1H), 3.76 (q, 2H), 1.07 (t, 3H).

[0103] Step 3: Preparation of Intermediate A1

[0104] DMF (500 mL) was added to the above intermediate A1-4 (56 g), and 19.2 g of sodium hydrogen hydride (60% W / W) and benzyl bromide (65.6 g) were added under stirring. The temperature was then raised to 60°C and the reaction was carried out for 10 hours. After the reaction was completed, the mixture was cooled to room temperature, and water (700 mL) was added to the reaction solution. The solid was precipitated, filtered, washed with 300 mL of water and then with 600 mL of n-heptane, and dried to obtain intermediate A1 (70.3 g, yield 83%).

[0105] ESI-MS (m / z): 264.8.

[0106] 1H-NMR(400MHz,DMSO-d6,δ / ppm):7.26-7.31(5H,m),6.03(1H,s),4.20(2H,s),3.66(q,2H),1.06(t,3H).

[0107] Intermediate A2: Preparation of 1-benzyl-6-chloro-3-propylpyrimidine-2,4(1H,3H)-dione

[0108] The synthetic route of intermediate A2 is as follows:

[0109]

[0110] Step 1: Preparation of intermediate A2-2

[0111] Add acetic acid (30 mL) to a three-necked flask, then add malonic acid (12 g) and N-propylurea (10 g), heat to 75°C, add acetic anhydride (20 mL) dropwise, continue heating to 85°C after stirring, stir for 4 hours until the reaction is complete, cool to 50°C, concentrate the reaction solution under reduced pressure, then add ethanol (20 mL), cool the mixture to 0°C, and stir for 2 hours. After filtering and drying, intermediate A2-2 (12.7 g, yield 75%) was obtained.

[0112] ESI-MS (m / z): 170.2.

[0113] 1H-NMR (400MHz, CDCl 3,δ / ppm):8.98(br,lH),3.79(t,2H),3.62(s,2H),1.65(m,2H),0.91(t,3H).

[0114] Step 2: Preparation of intermediate A2-3

[0115] Add POCl to the intermediate A2-2 (8.5 g). 3 (20 mL), water (0.5 mL) was added dropwise to the suspension at 0°C, and then the reaction mixture was heated at 80°C for 6 hours. The resulting brown solution was slowly cooled to room temperature, and POCl 3 The residue was washed with 50 mL of methanol and the obtained solid was recrystallized from ethanol to obtain Intermediate A2-3 (6.4 g, yield 68%).

[0116] ESI-MS (m / z): 188.7.

[0117] 1H-NMR (400MHz, CDCl 3 ,δ / ppm):11.09(s,1H),5.86(s,1H),3.82(t,2H),1.62(m,2H),0.94(t,3H).

[0118] Step 3: Preparation of Intermediate A2

[0119] DMF (450 mL) was added to the above intermediate A2-3 (5 g), and 1.6 g of sodium hydrogen hydride (60% W / W) and benzyl bromide (5.4 g) were added under stirring. The temperature was then raised to 65°C for reaction for 8 hours. After the reaction was completed, the mixture was cooled to room temperature, and water (50 mL) was added to the reaction solution. The solid was precipitated, filtered, washed with 20 mL of water and then with 50 mL of n-heptane, and dried to obtain intermediate A2 (5.8 g, yield 79%).

[0120] ESI-MS (m / z): 278.8.

[0121] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 7.27-7.33(5H,m),6.04(1H,s),4.21(2H,s),3.84(t,2H),1.63(m,2H),0.96(t,3H).

[0122] Intermediate A3: Preparation of 1-benzyl-6-chloro-3-cyclopropylpyrimidine-2,4(1H,3H)-dione

[0123] The synthetic route of intermediate A3 is as follows:

[0124]

[0125] Step 1: Preparation of intermediate A3-2

[0126] Add acetic acid (50 mL) to a three-necked flask, then add malonic acid (18 g) and N-cyclopropylurea (15 g), heat to 80°C, drop acetic anhydride (35 mL), stir and continue heating to 90°C, stir for 2 hours until the reaction is complete, cool to 50°C, concentrate the reaction solution under reduced pressure, then add ethanol (30 mL), cool the mixture to 0°C, and stir for 2 hours. After filtering and drying, intermediate A3-2 (18.2 g, yield 72%) was obtained.

[0127] ESI-MS (m / z): 168.2.

[0128] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 11.34 (s, 1H), 3.63 (s, 2H), 1.98 (m, 1H), 1.03 (dd, 2H), 0.87–0.68 (m, 2H).

[0129] Step 2: Preparation of intermediate A3-3

[0130] Add POCl to the intermediate A3-2 3 (80 mL), water (1.8 mL) was added dropwise to the suspension at 0°C, and then the reaction mixture was heated at 75°C for 5 hours. The resulting brown solution was slowly cooled to room temperature, and POCl 3 The residue was washed with 60 mL of methanol and the obtained solid was recrystallized from ethanol to obtain Intermediate A3-3 (14 g, yield 75%).

[0131] ESI-MS (m / z): 186.7.

[0132] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 11.36 (s, 1H), 5.93 (s, 1H), 1.99 (m, 1H), 1.05 (dd, 2H), 0.88–0.69 (m, 2H).

[0133] Step 3: Preparation of intermediate A3

[0134] DMF (100 mL) was added to the above intermediate A3-3 (9.3 g), and 2.8 g of sodium hydrogen sulfide (60% W / W) and benzyl bromide (9.8 g) were added under stirring. The temperature was then raised to 60°C and the reaction was carried out for 10 hours. After the reaction was completed, the mixture was cooled to room temperature, and water (50 mL) was added to the reaction solution. The solid was precipitated, filtered, washed with 50 mL of water and then with 100 mL of n-heptane, and dried to obtain intermediate A3 (12.2 g, yield 88%).

[0135] ESI-MS (m / z): 276.8.

[0136] 1H-NMR(400MHz,DMSO-d6,δ / ppm):7.25-7.31(5H,m),6.02(1H,s),4.15(2H,s),2.00(m,1H),1.07(dd,2H),0.89–0.70(m,2H).

[0137] Intermediate A4: Preparation of 1-benzyl-6-chloro-3-isopropylpyrimidine-2,4(1H,3H)-dione

[0138] The synthetic route of intermediate A4 is as follows:

[0139]

[0140] Step 1: Preparation of intermediate A4-2

[0141] Add acetic acid (50 mL) to a three-necked flask, then add malonic acid (17.2 g) and N-isopropylurea (15.3 g), heat to 75°C, drop acetic anhydride (35 mL), stir and continue heating to 90°C, stir for 3 hours until the reaction is complete, cool to 50°C, concentrate the reaction solution under reduced pressure, then add ethanol (30 mL), cool the mixture to 0°C, and stir for 2 hours. After filtration and drying, intermediate A4-2 (18.9 g, yield 74%) was obtained.

[0142] ESI-MS (m / z): 170.2.

[0143] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 11.19 (s, 1H), 4.83 (m, 1H), 3.58 (s, 2H), 1.32 (d, 6H).

[0144] Step 2: Preparation of intermediate A4-3

[0145] Add POCl to the intermediate A4-2 (17 g). 3 (100 mL), water (2 mL) was added dropwise to the suspension at 0°C, and the reaction mixture was heated at 85°C for 4 hours. The resulting brown solution was slowly cooled to room temperature, and POCl 3 The residue was washed with 50 mL of methanol and the obtained solid was recrystallized from ethanol to obtain Intermediate A4-3 (14.2 g, yield 76%).

[0146] ESI-MS (m / z): 188.7.

[0147] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 12.22 (s, 1H), 5.88 (s, 1H), 4.95 (m, 1H), 1.34 (d, 6H).

[0148] Step 3: Preparation of intermediate A4

[0149] DMF (150 mL) was added to the above intermediate A4-3 (14 g), and 4.3 g of sodium hydrogen hydride (60%) and benzyl bromide (14.5 g) were added under stirring. The temperature was then raised to 65°C for reaction for 12 hours. After the reaction was completed, the mixture was cooled to room temperature, and water (200 mL) was added to the reaction solution. The solid precipitated was filtered, washed with 80 mL of water and then with 120 mL of n-heptane, and dried to obtain intermediate A (16.1 g, yield 78%).

[0150] ESI-MS (m / z): 278.8.

[0151] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 7.26-7.39(5H,m),6.08(1H,s),4.98(m,1H),4.11(2H,s),1.38(d,6H).

[0152] Intermediate A6: Preparation of 1-benzyl-6-chloro-3-isobutylpyrimidine-2,4(1H,3H)-dione

[0153] The synthetic route of intermediate A6 is as follows:

[0154]

[0155] Step 1: Preparation of intermediate A6-2

[0156] Add acetic acid (30 mL) to a three-necked flask, then add malonic acid (13 g) and N-isobutylurea (11.6 g), heat to 70°C, drop acetic anhydride (25 mL), stir and continue heating to 90°C, stir for 3 hours until the reaction is complete, cool to 50°C, concentrate the reaction solution under reduced pressure, then add ethanol (20 mL), cool the mixture to 0°C, and stir for 2 hours. After filtration and drying, intermediate A6-2 (12.5 g, yield 68%) was obtained.

[0157] ESI-MS (m / z): 184.2.

[0158] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 11.43 (s, 1H), 3.68 (d, 2H), 3.64 (s, 2H), 1.94 (m, 1H), 0.94 (d, 6H).

[0159] Step 2: Preparation of intermediate A6-3

[0160] Add POCl to the intermediate A6-2 (9.2 g). 3 (50 mL), water (1.2 mL) was added dropwise to the suspension at 0°C, and then the reaction mixture was heated at 85°C for 3 hours. The resulting brown solution was slowly cooled to room temperature, and POCl 3 The residue was washed with 50 mL of methanol and the obtained solid was recrystallized from ethanol to obtain intermediate A6-3 (7 g, yield 70%).

[0161] ESI-MS (m / z): 202.7.

[0162] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 12.36 (s, 1H), 5.83 (s, 1H), 3.72 (d, 2H), 1.90 (m, 1H), 0.97 (d, 6H).

[0163] Step 3: Preparation of intermediate A6

[0164] DMF (60 mL) was added to the above intermediate A6-3 (6 g), and 1.8 g of sodium hydrogen hydride (60%) and benzyl bromide (6 g) were added under stirring. The temperature was then raised to 65°C for reaction for 8 hours. After the reaction was completed, the mixture was cooled to room temperature, and water (60 mL) was added to the reaction solution. The solid was precipitated, filtered, washed with 50 mL of water and then with 60 mL of n-heptane, and dried to obtain intermediate A6 (7.3 g, yield 85%).

[0165] ESI-MS (m / z): 292.8.

[0166] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 7.24-7.38(5H,m),6.10(1H,s),4.16(2H,s),3.72(d,2H),1.91(m,1H),0.94(d,6H).

[0167] Example 1: Preparation of (S)-3-ethyl-6-(1-phenylethoxy)pyrimidine-2,4(1H,3H)-dione (Compound 1)

[0168] The synthetic route of target compound 1 is as follows:

[0169]

[0170] Step 1: Preparation of intermediate B1-1

[0171] DMF (500 mL) was measured and placed in a three-necked flask, and the temperature was controlled to stir at 0°C. 18 g of sodium hydrogen sulfide (60%) and S-(-)-phenylethanol (40.3 g) were added under stirring, and the mixture was stirred for 1 hour. The above intermediate A1 (79.4 g) was then added thereto, and the mixture was heated to 80°C and reacted for 4 hours. After the reaction was completed, the mixture was cooled to room temperature, and 500 mL of water was added to the reaction solution to precipitate a solid. The solid was filtered, washed with water and then with n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 as eluent) to obtain intermediate B1-1 (79.8 g, yield 76%).

[0172] ESI-MS (m / z): 350.5.

[0173] 1H-NMR (400MHz, DMSO-d6, δ / ppm): δ7.38–7.24(m,10H),6.05(1H,s),4.29(q,1H),4.18(2H,s),3.71(q,2H),1.32(d,3H),1.06(t,3H).

[0174] Step 2: Preparation of compound 1

[0175] The intermediate B1-1 (35 g), palladium carbon (3 g, 10% W / W), and ethanol (400 mL) were added to the reaction bottle, replaced and controlled, connected to hydrogen, stirred and reacted at 35°C for 12 hours, and after the reaction, cooled to room temperature, filtered, concentrated under reduced pressure, washed with water and then with n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:5 as eluent) to obtain (S)-3-ethyl-6-(1-phenylethoxy)pyrimidine-2,4(1H,3H)-dione (25 g, yield 95%).

[0176] ESI-MS (m / z): 260.4.

[0177] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 11.40 (s, 1H), δ 7.35–7.20 (m, 5H), 6.03 (1H, s), 4.26 (q, 1H), 3.67 (q, 2H), 1.26 (d, 3H), 1.04 (t, 3H).

[0178] Example 2: Preparation of (S)-6-(1-phenylethoxy)-3-propylpyrimidine-2,4(1H,3H)-dione (Compound 2)

[0179] The synthetic route of target compound 2 is as follows:

[0180]

[0181] Step 1: Preparation of intermediate B2-1

[0182] DMF (250 mL) was measured and placed in a three-necked flask, the temperature was controlled to be stirred at 0°C, 1.1 g of sodium hydrogen hydride (60%) and S-(-)-phenylethanol (2.4 g) were added under stirring, and the mixture was stirred for 1 hour. The above intermediate A2 (5 g) was then added thereto, and the mixture was heated to 85°C and reacted for 5 hours. After the reaction was completed, the mixture was cooled to room temperature, water (10 mL) was added to the reaction solution, and the solid was precipitated. The solid was filtered, washed with 10 ml of water and then with 20 ml of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 as eluent) to obtain intermediate B2-1 (5.1 g, yield 77%).

[0183] ESI-MS (m / z): 364.5.

[0184] 1H-NMR (400MHz, DMSO-d6, δ / ppm): δ7.40–7.25(m,10H),6.05(1H,s),4.29 (q,1H),4.18(2H,s),3.84(t,2H),1.63(m,2H),1.32(d,3H),0.96(t,3H).

[0185] Step 2: Preparation of compound 2

[0186] The above intermediate B2-1 (3.5 g), 0.2 g palladium carbon (10%), and ethanol (20 mL) were added to a reaction bottle, replaced and controlled, connected to hydrogen, and stirred at 35°C for 10 hours. After the reaction, it was cooled to room temperature, filtered, concentrated under reduced pressure, washed with water and then with n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:4 as eluent) to obtain (S)-6-(1-phenylethoxy)-3-propylpyrimidine-2,4(1H,3H)-dione (2.4 g, yield 93%).

[0187] ESI-MS (m / z): 274.4.

[0188] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 11.42 (s, 1H), δ7.34–7.21 (m, 5H), 6.05 (1H,s),4.28(q,1H),3.86(t,2H),1.65(m,2H),1.33(d,3H),0.98(t,3H).

[0189] Example 3: Preparation of (S)-3-cyclopropyl-6-(1-phenylethoxy)pyrimidine-2,4(1H,3H)-dione (Compound 3)

[0190] The synthetic route of target compound 3 is as follows:

[0191]

[0192] Step 1: Preparation of intermediate B3-1

[0193] DMF (80 mL) was measured and placed in a three-necked flask, the temperature was controlled to stir at 0°C, 1.8 g of sodium hydrogen hydride (60% W / W) and S-(-)-phenylethanol (4 g) were added under stirring, and the mixture was stirred for 1 hour. The above intermediate A3 (8.3 g) was then added thereto, and the mixture was heated to 80°C and reacted for 4 hours. After the reaction was completed, the mixture was cooled to room temperature, water (40 mL) was added to the reaction solution, and the solid was precipitated. The solid was filtered, washed with water and then with n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 as eluent) to obtain intermediate B3-1 (8.8 g, yield 81%).

[0194] ESI-MS (m / z): 362.5.

[0195] 1H-NMR (400MHz, DMSO-d6, δ / ppm): δ7.38–7.21(m,10H),6.03(1H,s),4.27(q, 1H),4.15(2H,s),1.98(m,1H),1.31(d,3H),1.03(dd,2H),0.84–0.68(m,2H).

[0196] Step 2: Preparation of compound 3

[0197] The above intermediate B3-1 (8 g), 1 g palladium carbon (10% W / W), and ethanol (50 mL) were added to a reaction bottle, replaced and controlled, connected to hydrogen, and stirred at 35°C for 12 hours. After the reaction, it was cooled to room temperature, filtered, concentrated under reduced pressure, washed with 20 mL of water and then with 20 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:5 as eluent) to obtain (S)-3-cyclopropyl-6-(1-phenylethoxy)pyrimidine-2,4(1H,3H)-dione (5.6 g, yield 94%).

[0198] ESI-MS (m / z): 272.4.

[0199] 1H-NMR(400MHz,DMSO-d6,δ / ppm):11.37(s,1H),δ7.31–7.17(m,5H),6.03(1H ,s),4.22(q,1H),1.93(m,1H),1.29(d,3H),1.02(dd,2H),0.85–0.70(m,2H).

[0200] Example 4: Preparation of (S)-3-isopropyl-6-(1-phenylethoxy)pyrimidine-2,4(1H,3H)-dione (Compound 4)

[0201] The synthetic route of target compound 4 is as follows:

[0202]

[0203] Step 1: Preparation of intermediate B4-1

[0204] DMF (120 mL) was measured and placed in a three-necked flask, the temperature was controlled to stir at 0°C, 2.4 g of sodium hydrogen hydride (60% W / W) and S-(-)-phenylethanol (6.4 g) were added under stirring, and the mixture was stirred for 1 hour. The above intermediate A4 (13.9 g) was then added thereto, and the mixture was heated to 85°C and reacted for 4 hours. After the reaction was completed, the mixture was cooled to room temperature, water (80 mL) was added to the reaction solution, and a solid was precipitated. The solid was filtered, washed with 50 mL of water and then with 60 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:1 as eluent) to obtain intermediate B4-1 (13.4 g, yield 74%).

[0205] ESI-MS (m / z): 364.5.

[0206] 1H-NMR (400MHz, DMSO-d6, δ / ppm): δ7.38–7.21(m,10H),6.03(1H,s),4.98(m,1H),4.27(q,1H),4.16(2H,s),1.35(d,6H),1.30(d,3H).

[0207] Step 2: Preparation of compound 4

[0208] The above intermediate B4-1 (6.5 g), 0.5 g palladium carbon (10% W / W), and ethanol (50 mL) were added to a reaction bottle, replaced and controlled, connected to hydrogen, and stirred at 35°C for 12 hours. After the reaction, it was cooled to room temperature, filtered, concentrated under reduced pressure, washed with 20 mL of water and then with 20 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 as eluent) to obtain (S)-3-isopropyl-6-(1-phenylethoxy)pyrimidine-2,4(1H,3H)-dione (4.5 g, yield 92%).

[0209] ESI-MS (m / z): 274.4.

[0210] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 11.34 (s, 1H), δ 7.30–7.18 (m, 5H), 6.02 (1H, s), 4.93 (m, 1H), 4.28 (q, 1H), 1.33 (d, 6H), 1.34 (d, 3H).

[0211] Example 5: Preparation of (S)-3-isobutyl-6-(1-phenylethoxy)pyrimidine-2,4(1H,3H)-dione (Compound 6)

[0212] The synthetic route of target compound 6 is as follows:

[0213]

[0214] Step 1: Preparation of intermediate B6-1

[0215] DMF (40 mL) was measured and placed in a three-necked flask, the temperature was controlled to be stirred at 0°C, 1 g of sodium hydrogen hydride (60% W / W) and S-(-)-phenylethanol (2.5 g) were added under stirring, and the mixture was stirred for 1 hour. The above intermediate A6 (6 g) was then added thereto, and the mixture was heated to 85°C and reacted for 5 hours. After the reaction was completed, the mixture was cooled to room temperature, water (40 mL) was added to the reaction solution, and the solid was precipitated. The solid was filtered, washed with 20 mL of water and then with 30 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:1 as eluent) to obtain intermediate B6-1 (5.7 g, yield 73%).

[0216] ESI-MS (m / z): 378.5.

[0217] 1H-NMR (400MHz, DMSO-d6, δ / ppm): δ7.35–7.10(m,10H),6.15(1H,s),4.25 (q,1H),4.18(2H,s),3.75(d,2H),1.96(m,1H),1.32(d,3H),0.95(d,6H).

[0218] Step 2: Preparation of compound 6

[0219] The above intermediate B6-1 (3 g), palladium carbon (0.3 g, 10% W / W), and ethanol (15 mL) were added to a reaction bottle, replaced and controlled, connected to hydrogen, and stirred at 35°C for 12 hours. After the reaction, it was cooled to room temperature, filtered, concentrated under reduced pressure, washed with 10 mL of water and then with 20 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 as eluent) to obtain (S)-3-isobutyl-6-(1-phenylethoxy)pyrimidine-2,4(1H,3H)-dione (2.1 g, yield 90%).

[0220] ESI-MS (m / z): 288.4.

[0221] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 11.32 (s, 1H), δ7.28–7.15 (m, 5H), 6.08 (1H,s),4.23(q,1H),3.95(d,2H),1.94(m,1H),1.41(d,3H),0.98(d,6H).

[0222] Example 6: Preparation of (S)-3-ethyl-6-(1-(3-fluorophenyl)ethoxy)pyrimidine-2,4(1H,3H)-dione (Compound 9)

[0223] The synthetic route of target compound 9 is as follows:

[0224]

[0225] Step 1: Preparation of intermediate B9-1

[0226] DMF (40 mL) was measured and placed in a three-necked flask, the temperature was controlled to be stirred at 0°C, 1.2 g of sodium hydrogen hydride (60% W / W) and (S)-1-(3-fluorophenyl)ethanol (2.8 g) were added under stirring, and the mixture was stirred for 1 hour. The above intermediate A1 (5.3 g) was then added thereto, and the temperature was raised to 80°C for reaction for 4 hours. After the reaction was completed, the mixture was cooled to room temperature, water (50 mL) was added to the reaction solution, and the solid was precipitated. The solid was filtered, washed with 10 mL of water and then with 10 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 as eluent) to obtain intermediate B9-1 (5.3 g, yield 72%).

[0227] ESI-MS (m / z): 368.5.

[0228] 1H-NMR (400MHz, DMSO-d6, δ / ppm): δ7.38(m,1H), δ7.28–7.17(m,5H),7.12-6.95(m, 3H),5.42(1H,s),4.36(q,1H),4.26(2H,s),3.62(q,2H),1.43(d,3H),1.16(t,3H).

[0229] Step 2: Preparation of compound 9

[0230] The above intermediate B9-1 (3.5 g), 0.5 g palladium carbon (10% W / W), and ethanol (40 mL) were added to a reaction bottle, replaced and controlled, connected to hydrogen, and stirred at 35°C for 12 hours. After the reaction, it was cooled to room temperature, filtered, concentrated under reduced pressure, washed with 10 mL of water and then with 10 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:3 as eluent) to obtain (S)-3-ethyl-6-(1-(3-fluorophenyl)ethoxy)pyrimidine-2,4(1H,3H)-dione (2.4 g, yield 92%).

[0231] ESI-MS (m / z): 278.4.

[0232] 1H-NMR(400MHz,DMSO-d6,δ / ppm):11.31(s,1H),δ7.28(m,1H),7.02-6.85 (m,3H),5.47(1H,s),4.21(q,1H),3.53(q,2H),1.31(d,3H),1.12(t,3H).

[0233] Example 7: Preparation of (S)-3-ethyl-6-(1-(3-chlorophenyl)ethoxy)pyrimidine-2,4(1H,3H)-dione (Compound 11)

[0234] The synthetic route of target compound 11 is as follows:

[0235]

[0236] Step 1: Preparation of intermediate B11-1

[0237] DMF (20 mL) was measured and placed in a three-necked flask, the temperature was controlled to be stirred at 0°C, 0.6 g of sodium hydrogen sulfide (60% W / W) and (S)-1-(3-chlorophenyl)ethanol (1.5 g) were added under stirring, and the mixture was stirred for 1 hour. The above intermediate A2 (2.8 g) was then added thereto, and the mixture was heated to 80°C for reaction for 5 hours. After the reaction was completed, the mixture was cooled to room temperature, water (20 mL) was added to the reaction solution, and the solid was precipitated. The solid was filtered, washed with 20 mL of water and then with 30 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 as eluent) to obtain intermediate B11-1 (3.1 g, yield 78%).

[0238] ESI-MS (m / z): 398.9.

[0239] 1H-NMR (400MHz, DMSO-d6, δ / ppm): δ7.43(m,1H),7.28–7.17(m,5H),7.22-7.06(m,3H),5. 65(1H,s),4.24(q,1H),4.28(2H,s),3.91(t,2H),1.67(m,2H),1.35(d,3H),0.93(t,3H).

[0240] Step 2: Preparation of compound 11

[0241] The above intermediate B11-1 (3 g), palladium carbon (10% W / W, 0.3 g), and ethanol (20 mL) were added to a reaction bottle, and the mixture was replaced and connected with hydrogen. The mixture was stirred at 35°C for 12 hours. After the reaction, the mixture was cooled to room temperature, filtered, concentrated under reduced pressure, washed with 10 mL of water and then with 10 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:3 as eluent) to obtain (S)-3-ethyl-6-(1-(3-chlorophenyl)ethoxy)pyrimidine-2,4(1H,3H)-dione (2.2 g, yield was 94%).

[0242] ESI-MS (m / z): 308.8.

[0243] 1H-NMR(400MHz,DMSO-d6,δ / ppm):11.52(s,1H),δ7.47(m,1H),7.24-7.16(m,3H ),5.60(1H,s),4.22(q,1H),3.76(t,2H),1.64(m,2H),1.32(d,3H),0.95(t,3H).

[0244] Example 8: Preparation of (S)-3-isopropyl-6-(1-(3-fluorophenyl)ethoxy)pyrimidine-2,4(1H,3H)-dione (Compound 13)

[0245] The synthetic route of target compound 13 is as follows:

[0246]

[0247] Step 1: Preparation of intermediate B13-1

[0248] DMF (30 mL) was measured and placed in a three-necked flask, and the temperature was controlled to stir at 0°C. 0.7 g of sodium hydrogen sulfide (60% W / W) and (S)-1-(3-fluorophenyl)ethanol (1.4 g) were added under stirring, and the mixture was stirred for 1 hour. The above intermediate A4 (2.8 g) was then added thereto, and the mixture was heated to 80°C and reacted for 4 hours. After the reaction was completed, the mixture was cooled to room temperature, 30 mL of water was added to the reaction solution, and a solid was precipitated. The solid was filtered, washed with 15 mL of water and then with 20 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 as eluent) to obtain intermediate B13-1 (2.5 g, yield 66%).

[0249] ESI-MS (m / z): 382.5.

[0250] 1H-NMR (400MHz, DMSO-d6, δ / ppm): δ7.39(m,1H),7.32–7.27(m,5H),7.14-6.85(m, 3H),5.45(1H,s),4.83(m,1H),4.21(q,1H),4.12(2H,s),1.33(d,6H),1.20(d,3H).

[0251] Step 2: Preparation of compound 13

[0252] The above intermediate B13-1 (2 g), palladium carbon (10% W / W, 0.3 g), and ethanol (20 mL) were added to a reaction bottle, replaced and controlled, connected to hydrogen, and stirred at 35°C for 12 hours. After the reaction, it was cooled to room temperature, filtered, concentrated under reduced pressure, washed with 10 mL of water and then with 10 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:5 as eluent) to obtain (S)-3-isopropyl-6-(1-(3-fluorophenyl)ethoxy)pyrimidine-2,4(1H,3H)-dione (1.4 g, yield 90%).

[0253] ESI-MS (m / z): 292.4.

[0254] 1H-NMR(400MHz,DMSO-d6,δ / ppm):11.28(s,1H),δ7.28(m,1H),7.12-6.93 (m,3H),5.35(1H,s),4.83(m,1H),4.21(q,1H),1.23(d,6H),1.14(d,3H).

[0255] Example 9: Preparation of (S)-3-isobutyl-6-(1-(3-(trifluoromethyl)phenyl)ethoxy)pyrimidine-2,4(1H,3H)-dione (Compound 14) The synthetic route of target compound 14 is as follows:

[0256]

[0257] Step 1: Preparation of intermediate B14-1

[0258] DMF (30 mL) was measured and placed in a three-necked flask, and the temperature was controlled to stir at 0°C. 0.7 g of sodium hydrogen sulfide (60% W / W) and (S)-1-(3-fluoromethylphenyl)ethanol (1.9 g) were added under stirring, and the mixture was stirred for 1 hour. The above intermediate A6 (3 g) was then added thereto, and the mixture was heated to 80°C and reacted for 5 hours. After the reaction was completed, the mixture was cooled to room temperature, 30 mL of water was added to the reaction solution, and the solid was precipitated. The solid was filtered, washed with 20 mL of water and then with 30 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 as eluent) to obtain intermediate B14-1 (3.5 g, yield 76%).

[0259] ESI-MS (m / z): 446.5.

[0260] 1H-NMR (400MHz, DMSO-d6, δ / ppm): δ7.58(m,1H),7.32-7.13(m,8H),5.65(1H,s) ,4.45(q,1H),4.28(2H,s),3.85(d,2H),1.93(m,1H),1.42(d,3H),0.91(d,6H).

[0261] Step 2: Preparation of compound 14

[0262] 3 g of the intermediate B14-1, 0.4 g of palladium carbon (10% W / W) and ethanol (20 mL) were added to a reaction bottle, and the mixture was replaced and connected with hydrogen. The mixture was stirred and reacted at 35°C for 12 hours. After the reaction, the mixture was cooled to room temperature, filtered, concentrated under reduced pressure, washed with 20 mL of water and then with 20 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:4 as eluent) to obtain (S)-3-isobutyl-6-(1-(3-(trifluoromethyl)phenyl)ethoxy)pyrimidine-2,4(1H,3H)-dione (2.2 g, yield 92%).

[0263] ESI-MS (m / z): 356.4.

[0264] 1H-NMR(400MHz,DMSO-d6,δ / ppm):11.62(s,1H),δ7.68(m,1H),7.42-7.23(m,3H ),5.75(1H,s),4.13(q,1H),3.75(d,2H),1.84(m,1H),1.45(d,3H),0.88(d,6H).

[0265] Example 10: Preparation of (S)-6-(1-(1H-indol-5-yl)ethoxy)-3-isopropylpyrimidine-2,4(1H,3H)-dione (Compound 19)

[0266] The synthetic route of target compound 19 is as follows:

[0267]

[0268] Step 1: Preparation of intermediate B19-1

[0269] DMF (30 mL) was measured and placed in a three-necked flask, the temperature was controlled to stir at 0°C, 0.7 g of sodium hydrogen hydride (60% W / W) and 1.6 g of (S)-1-(1H-indol-5-yl)ethanol were added under stirring, and the mixture was stirred for 1 hour. The above intermediate A4 (2.8 g) was then added thereto, and the mixture was heated to 85°C and reacted for 4 hours. After the reaction was completed, the mixture was cooled to room temperature, water (30 mL) was added to the reaction solution, and a solid was precipitated. The solid was filtered, washed with 20 mL of water and then with 30 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 as eluent) to obtain intermediate B19-1 (2.6 g, yield 64%).

[0270] ESI-MS (m / z): 403.5.

[0271] 1H-NMR(400MHz,DMSO-d6,δ / ppm):10.95(s,1H),7.34–7.21(m,8H),6.95(dd,1H),6.35–6 .37(m,1H),6.03(1H,s),4.98(m,1H),4.27(q,1H),4.16(2H,s),1.35(d,6H),1.30(d,3H).

[0272] Step 2: Preparation of compound 19

[0273] The above intermediate B19-1, palladium carbon (10% W / W, 0.3 g) and ethanol (30 mL) were added to a reaction bottle, replaced and controlled, connected to hydrogen, and stirred at 35°C for 12 hours. After the reaction, it was cooled to room temperature, filtered, concentrated under reduced pressure, washed with 10 mL of water and then with 10 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:3 as eluent) to obtain (S)-6-(1-(1H-indol-5-yl)ethoxy)-3-isopropylpyrimidine-2,4(1H,3H)-dione (1.6 g, yield 85%).

[0274] ESI-MS (m / z): 313.4.

[0275] 1H-NMR(400MHz,DMSO-d6,δ / ppm):11.35(s,1H),11.05(s,1H),7.54–7.46(m,1H),7.32–7.23(m,2H) ,6.98(dd,1H),6.45–6.39(m,1H),6.02(1H,s),4.93(m,1H),4.28(q,1H),1.33(d,6H),1.34(d,3H).

[0276] Example 11: Preparation of (S)-6-(1-(Benzofuran-5-yl)ethoxy)-3-isopropylpyrimidine-2,4(1H,3H)-dione (Compound 20) The synthetic route of target compound 20 is as follows:

[0277]

[0278] Step 1: Preparation of intermediate B20-1

[0279] DMF (25 mL) was measured and placed in a three-necked flask, the temperature was controlled to stir at 0°C, 0.7 g of sodium hydrogen sulfide (60% W / W) and (S)-1-(benzofuran-5-yl)ethanol (1.6 g) were added under stirring, and the mixture was stirred for 1 hour. The above intermediate A4 (2.8 g) was then added thereto, and the mixture was heated to 85°C and reacted for 5 hours. After the reaction was completed, the mixture was cooled to room temperature, 30 mL of water was added to the reaction solution, a solid was precipitated, and the solid was filtered, washed with 20 mL of water and then with 30 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:3 as eluent) to obtain intermediate B20-1 (2.5 g, yield 62%).

[0280] ESI-MS (m / z): 404.5.

[0281] 1H-NMR (400MHz, DMSO-d6, δ / ppm): δ7.97-7.94(m,1H),7.60(s,1H),7.50(d,1H),7.29-7.27(m,6H) ,6.94-6.91(m,1H),6.03(1H,s),4.98(m,1H),4.27(q,1H),4.16(2H,s),1.35(d,6H),1.30(d,3H).

[0282] Step 2: Preparation of compound 20

[0283] The above intermediate B20-1 (2 g), 0.2 g palladium carbon (10% W / W), and ethanol (400 mL) were added to a reaction bottle, replaced and controlled, connected to hydrogen, and stirred at 35°C for 12 hours. After the reaction, it was cooled to room temperature, filtered, concentrated under reduced pressure, washed with 10 mL of water and then with 20 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:5 as eluent) to obtain (S)-6-(1-(benzofuran-5-yl)ethoxy)-3-isopropylpyrimidine-2,4(1H,3H)-dione (1.3 g, yield 86%).

[0284] ESI-MS (m / z): 314.4.

[0285] 1H-NMR(400MHz,DMSO-d6,δ / ppm):11.45(s,1H),7.93-7.84(m,1H),7.64(s,1H),7.53(d,1H) ,7.23(m,1H),6.984(m,1H),6.06(1H,s),4.84(m,1H),4.21(q,1H),1.32(d,6H),1.36(d,3H).

[0286] Example 12: Preparation of (S)-6-(1-(benzo[d][1,3]dioxol-5-yl)ethoxy)-3-isopropylpyrimidine-2,4(1H,3H)-dione (Compound 22) The synthetic route of target compound 22 is as follows:

[0287]

[0288] Step 1: Preparation of intermediate B22-1

[0289] DMF (30 mL) was measured and placed in a three-necked flask, the temperature was controlled to be stirred at 0°C, 0.7 g of sodium hydride (60%) and (S)-1-(benzo[d][1,3]dioxol-5-yl)ethanol (1.7 g) were added under stirring, and the mixture was stirred for 1 hour. The intermediate A4 (2.8 g) was then added thereto, and the temperature was raised to 85°C for reaction for 4 hours. After the reaction was completed, the mixture was cooled to room temperature, water (30 mL) was added to the reaction solution, and the solid was precipitated. The solid was filtered, washed with 20 mL of water and then with 30 mL of n-heptane, and purified by column chromatography (petroleum ether:ethyl acetate = 1:2 as eluent) to obtain intermediate B22-1 (2.9 g, yield 72%).

[0290] ESI-MS (m / z): 408.5.

[0291] 1H-NMR (400MHz, DMSO-d6, δ / ppm): δ7.24-7.21(m,5H),6.75(d,1H),6.52(s,1H),6.30(d,1H) ,6.13(1H,s),5.91(s,2H),4.95(m,1H),4.34(q,1H),4.26(2H,s),1.37(d,6H),1.32(d,3H).

[0292] Step 2: Preparation of compound 22

[0293] The above intermediate B22-1 (2.7 g), 0.3 g palladium carbon (10%), and ethanol (40 mL) were added to a reaction bottle, replaced and controlled, connected to hydrogen, and stirred at 35°C for 12 hours. After the reaction, it was cooled to room temperature, filtered, concentrated under reduced pressure, washed with 20 mL of water and then with 20 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:4 as eluent) to obtain (S)-6-(1-(benzo[d][1,3]diol-5-yl)ethoxy)-3-isopropylpyrimidine-2,4(1H,3H)-dione (1.8 g, yield 85%).

[0294] ESI-MS (m / z): 318.4.

[0295] 1H-NMR(400MHz,DMSO-d6,δ / ppm):11.36(s,1H),6.78(d,1H),6.42(s,1H),6.34(d,1H),6.23(1H,

[0296] s),5.82(s,2H),4.87(m,1H),4.24(q,1H),1.35(d,6H),1.22(d,3H).

[0297] Example 13: Preparation of (S)-3-isopropyl-6-(1-(naphthalen-2-yl)ethoxy)pyrimidine-2,4(1H,3H)-dione (Compound 24)

[0298] The synthetic route of target compound 24 is as follows:

[0299]

[0300] Step 1: Preparation of intermediate B24-1

[0301] DMF (35 mL) was measured and placed in a three-necked flask, the temperature was controlled to stir at 0°C, 0.7 g of sodium hydrogen hydride (60% W / W) and (S)-1-(naphthalene-2-yl)ethanol (1.7 g) were added under stirring, and the mixture was stirred for 1 hour. The intermediate A4 (2.8 g) was then added thereto, and the mixture was heated to 85°C and reacted for 4 hours. After the reaction was completed, the mixture was cooled to room temperature, water (30 mL) was added to the reaction solution, a solid was precipitated, and the solid was filtered, washed with 20 mL of water and then with 30 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:1 as eluent) to obtain intermediate B24-1 (3.2 g, yield 77%).

[0302] ESI-MS (m / z): 414.5.

[0303] 1H-NMR(400MHz,DMSO-d6,δ / ppm):7.96(m,3H),7.89(s,1H),7.57(m,3H),7.24-7.21(m,5

[0304] H),6.23(1H,s),4.85(m,1H),4.31(q,1H),4.22(2H,s),1.33(d,6H),1.30(d,3H).

[0305] Step 2: Preparation of compound 24

[0306] The above intermediate B24-1 (3 g), 0.3 g palladium carbon (10% W / W), and ethanol (40 mL) were added to a reaction bottle, replaced and controlled, connected to hydrogen, and stirred at 35°C for 12 hours. After the reaction, it was cooled to room temperature, filtered, concentrated under reduced pressure, washed with 20 mL of water and then with 20 mL of n-heptane, and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 as eluent) to obtain (S)-3-isopropyl-6-(1-(naphthalen-2-yl)ethoxy)pyrimidine-2,4(1H,3H)-dione (2.2 g, yield 92%).

[0307] ESI-MS (m / z): 324.4.

[0308] 1H-NMR(400MHz,DMSO-d6,δ / ppm):11.42(s,1H),7.76(m,3H),7.62(s,1H),7 .40(m,3H),6.26(1H,s),4.77(m,1H),4.23(q,1H),1.32(d,6H),1.20(d,3H).

[0309] The following target compounds were prepared similarly to the synthesis method of compound 1.

[0310]

[0311]

[0312]

[0313] Test Example 1: Determination of myosin inhibitory activity

[0314] To evaluate the effect of the compounds of the invention on the ATP (adenosine triphosphatase) activity of full-length myometrial actomyosin in the context of native sarcomeres, a skinned myofibril analysis was performed. Rat cardiac myofibrils were obtained by homogenizing rat (SPF grade, 200-220 g, male) left ventricular tissue in the presence of a detergent (e.g., triton X-100). This treatment removes membranes and most soluble cytoplasmic proteins, but leaves the myometrial actomyosin structure intact. The myofibril preparation retains the Ca- 2+ The ability to hydrolyze ATP in a controlled manner. At a defined fraction (i.e., 25%, 75%) of the activation rate, Ca 2+ The myofibril preparations were analyzed for ATPase activity in the presence and absence of the compound at 100 μg / ml concentrations. The ability of the compounds to inhibit the steady-state ATPase activity of rat cardiac myofibrils was evaluated using a pyruvate kinase and lactate dehydrogenase (PK / LDH) coupled enzyme system. This assay regenerates the ADP produced by myosin into ATP by oxidizing NADH (nicotinamide adenine dinucleotide), thereby producing an absorbance change at 340 nm. Prior to testing the compounds, the calcium responsiveness of rat cardiac myofibrils was evaluated, and the calcium concentration that achieved 50% (pCa50) or 75% (pCa75) activation of the myofibril system was selected as the inhibitory activity condition for evaluating the compounds. All enzyme activities were measured in a pH 6.8 buffer solution containing 12 mM PIPES (piperazine-N,N'-bis(2-ethanesulfonic acid)), 2 mM magnesium chloride. The final assay conditions were 1 mg / mL rat cardiac myofibrils, 4 U / mL pyruvate kinase, 6 U / mL lactate dehydrogenase, 50 uM ATP, 0.1 mg / mL BSA (rat serum albumin), 10 ppm defoamer, 1 mM DTT (dithiothreitol), 0.5 mM NADH (nicotinamide adenine dinucleotide), 1.5 mM PEP, 0.6 mM EGTA, and an amount of CaCl sufficient to achieve 50% or 75% activation of myofibril ATPase activity. 2 .

[0315] The compound was diluted 3-fold with DMSO to 11 different concentrations, with the starting concentration being 0.1 mM. 2uL of the above compounds of different concentrations were added to a 96-well plate, 100uL of rat cardiac myofibrils were added, 40uL of a solution containing PK / LDH, BSA and DTT, and 40μL of a solution containing ATP, NADH and PEP were added, and finally 20uL of a calcium solution (the free calcium concentration required to achieve 50% activation of the myofibril system) was added to start the enzymatic reaction. At 25°C, M200 PRO-grating multifunctional microplate reader, read the absorbance at 340nm, read once every 30s, and read continuously for 15min. Record the data as the slope of the absorbance response to time. The slope of the absorbance response as a function of time is normalized to the slope on the plate containing DMSO. The normalized ratio is then plotted as a function of drug concentration, and the data is fitted into a curve using Graph PadPrism. The midpoint of the curve is the IC50, which is the drug concentration when 50% of the total response is inhibited.

[0316] The results of the tested compounds are shown in Table 1. The tested compounds were prepared according to the synthesis method described herein.

[0317] Table 1 Myosin inhibitory activity of the compounds of the present invention

[0318]

[0319] The experimental results show that the compounds disclosed herein have an inhibitory effect on myosin ATPase.

[0320] Test Example 2: Contractility measurement of adult rat cardiomyocytes

[0321] Adult male rats were anesthetized and the heart was rapidly removed, flushed, and the ascending aorta cannulated. 2 Continuous retrograde perfusion was started with a perfusion pressure of 1000 μg / mL. First, a nominal Ca-free 2+ Modified Krebs solution for perfusing the heart: 113mM

[0322] NaCl, 4.7 mM KCl, 0.6 mM KH 2 PO 4 , 0.6 mM Na 2 HPO 4 、1.2mMMgS0 4 , 12 mM NaHCO 3 、10mM

[0323] KHCO 3, 30 mM taurine, 5.5 mM glucose and 10 mM Hepes (Sigma). This medium was made with 95% O 2 -5% CO 2 The mixture was continuously aerated. The heart was perfused with modified Krebs buffer supplemented with collagenase and a final calcium concentration of 12.5 μM. After the heart appeared white and soft in appearance, the heart was removed from the cannula. The atria and blood vessels were removed, and the ventricles were gently cut into smaller pieces with forceps. The tissue was homogenized by repeated pipette trituration, and the collagenase reaction was stopped by 10% calf serum (BCS), precipitated and resuspended in 5% BCS and 12.5 μM CaCl 2 The perfusion buffer was filled with CaCl 2 The solution was added to a final concentration of 1.2 mM to make the myocytes calcium tolerant. The cells were then washed and resuspended in Tyrode's buffer (137 mM NaCl, 3.7 mM KCl, 0.5 mM

[0324] MgCl, 11 mM glucose, 4 mM Hepes, and 1.2 mM CaCl 2 , pH 7.4). Cells were kept at 37°C for 60 min before starting the experiment and used within 5 hours of separation. Cell preparations were used only if the cells first passed the QC standard by showing a contraction response to standard (>150% of basal) and isoproterenol (ISO; >250% of basal) treatment. In addition, only cells with a basal contraction rate between 3% and 8% were used in subsequent experiments with compounds.

[0325] An aliquot of myocytes in Tyrode's buffer was placed in a perfusion chamber equipped with a heating stage. Myocytes were allowed to attach, the chamber was heated to 37°C, and the cells were perfused with 37°C Tyrode's buffer. Myocytes were stimulated with a 1 Hz (20% above threshold) electric field using platinum electrodes. Only cells with clear striations and that were quiescent before pacing were used for contractility experiments. To determine basal contraction rate, myocytes were imaged via a 40x objective. Images were digitized using a variable frame rate (60 Hz-240 Hz) charge-coupled device camera and displayed on a computer screen (IonOptix Milton) at a sampling rate of 240 Hz. Once cell contraction stabilized over time, test compounds (0.01 μM-15 μM) were added.

[0326] Perfusion is carried out in the chamber of myocyte for 5 minutes.Then edge detection is used to record the contraction rate and contraction and relaxation speed of myocyte.IonOptix software is used to continuously record data.Ionwizard software (IonOptix) is used to analyze contractility data, and for each cell, 10-20 contractility transient states are averaged and the baseline (no compound) conditions are compared with the conditions treated with compounds.Compounds are measured by the effect of shortening fraction (FS), wherein the shortening fraction is the maximum length of the cell when contracted divided by the baseline cell length and the ratio normalized to 100% relative to untreated cells.

[0327] Table 2 Inhibition of cardiomyocyte contraction by selected compounds

[0328] Compound No. Activity at 0.5 μM Activity at 1.0 μM 1 + ++ 4 ++ +++ 19 ++ nd 21 nd nd 23 nd ++ 25 nd + MYK-461 ++ +++

[0329] + indicates that the shortening fraction inhibition value is less than 33%; ++ indicates that the shortening fraction inhibition value is between 33% and 66%; +++ indicates that the shortening fraction inhibition value is greater than 66%. nd indicates no inhibition.

[0330] The experimental results showed that at the same molar concentration, the inhibitory effect of compound 4 was equivalent to that of MYK-461.

[0331] Test Example 3: Pharmacokinetics of the compound in rats

[0332] Six male SD rats were divided into two groups, 3 rats in each group, and the test compound 4 was administered intravenously and orally at doses of 0.1 mg / kg and 1 mg / kg, respectively. The rats were fasted overnight before administration, and food was resumed 4 hours after administration. Water was freely available during the entire test period. After administration, 0.2-0.3 mL of blood was collected from the orbital venous plexus according to the time points (before administration, 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h and 24 h after administration), placed in an anticoagulated test tube containing 2 μL 20% EDTA-K2 and stored on wet ice. The supernatant plasma was obtained by centrifugation at 6000 rpm for 8 min at low temperature (4-10°C) within 1 hour, and stored in an ultra-low temperature freezer (below -60°C) for testing. The concentration of compound 4 in the plasma sample was detected by LC-MS / MS method, and the software Calculate pharmacokinetic parameters.

[0333] Table 3: Pharmacokinetic parameters of compound 4 in rats after oral administration

[0334]

[0335] The experimental results show that the pharmacokinetic absorption of compound 4 in SD rats is good. In addition, the T of compound 4 in the present invention is higher than that of compound MYK-461. 1 / 2The compound MYK-461 has a significant shortening due to T 1 / 2 It is relatively long, and the accumulation is serious in clinical practice. Clinical medication needs to be adjusted continuously, which increases the risk of medication. 1 / 2 , can reduce or avoid clinical drug accumulation in the body, which is beneficial to determine the dosage of patients and avoid the risk caused by accumulation. Obviously, the compound 4 disclosed in the present invention has obvious pharmacokinetic advantages over the compound MYK-461.

[0336] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A barbituric acid derivative of formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug: in, X1 and X2 are ring atoms and are each independently selected from N or C; Group fragment wherein A is absent, and X1 and X2 are independently substituted by R5 and R6; or when A is present, A, together with the ring atoms of X1 and X2, form a 3-10 membered heterocycloalkyl, a 6-10 membered aryl or a 5-10 membered heteroaryl, wherein the heteroatom is independently selected from at least one of N, O and S; and A is further substituted by one or more R a Replace, when R a When it is multiple, the R a Same or different; R a R5 and R6 are each independently selected from H, halogen, hydroxyl, amino, cyano, oxo, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C3-C8 halocycloalkyl and C1-C6 haloalkoxy; the C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C3-C8 halocycloalkyl and C1-C6 haloalkoxy are optionally substituted by one or more R b Substitution; said R b A substituent selected from the group consisting of H, halogen, hydroxy, amino, nitro, cyano, oxo, carboxyl, C1-C6 alkyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy; when R b When it is multiple, the R b Same or different; R1 is selected from H, halogen, C1-C6 alkyl, C2-C6 alkenyl, C3-C8 cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aryl and 5-10 membered heteroaryl, wherein the C1-C6 alkyl, C2-C6 alkenyl, C3-C8 cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aryl and 5-10 membered heteroaryl are optionally replaced by one or more R c Substitution; said R c A substituent selected from the group consisting of H, halogen, hydroxy, amino, nitro, cyano, oxo, carboxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy; when R c When it is multiple, the R c Same or different; R2 is selected from H and C1-C6 alkyl; R3 is selected from H, halogen, hydroxy, amino, nitro, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy, wherein the C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy are optionally replaced by one or more R d Substitution; said R d A substituent selected from the group consisting of H, halogen, hydroxy, amino, nitro, cyano, oxo, carboxyl, C1-C6 alkyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy; when R d When it is multiple, the R d Same or different; m and n are 1, 2 or 3 respectively.

2. The barbituric acid derivative of formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug according to claim 1, characterized in that: It has the structure shown in formula II: Wherein, X1, X2, A, R1, R2, R3 and n are defined as in claim 1.

3. The barbituric acid derivative of formula I according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that: The barbituric acid derivative is any of the following: Scheme 1: The barbituric acid derivative has a structure as shown in the following formula I-1: in, R5 and R6 are each independently selected from H, halogen, hydroxy, amino, cyano, oxo, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C3-C8 halocycloalkyl and C1-C6 haloalkoxy; the C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C3-C8 halocycloalkyl and C1-C6 haloalkoxy are optionally substituted by one or more R b replace; R1, R2, R3, R b , m and n are as defined in claim 1; Scheme 2: The barbituric acid derivative has a structure as shown in the following formula I-2: in, A and the ring atoms X1 and X2 together form a 3-10-membered heterocycloalkyl, a 6-10-membered aryl or a 5-10-membered heteroaryl, wherein the heterocyclic ring is independently selected from at least one of N, O and S; and A is further substituted by one or more R a Replace, when R a When it is multiple, the R a Same or different; R1, R2, R3, R a , m and n are as defined in claim 1.

4. The barbituric acid derivative of formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug according to claim 1, characterized in that: Group fragment In the formula (A), A exists and A, together with the ring atoms X1 and X2, forms a 3- to 8-membered heterocycloalkyl, a 6- to 8-membered aryl, or a 5- to 8-membered heteroaryl; Optionally, the 3- to 8-membered heterocycloalkyl, 6- to 8-membered aryl or 5- to 8-membered heteroaryl has 1 to 3 heteroatoms selected from N, O and S; Preferably, the group fragment In the formula, A, together with the ring atoms X1 and X2, forms 5. The barbituric acid derivative of formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug according to claim 1, characterized in that:R1 is selected from C1-C6 alkyl, C2-C6 alkenyl, C3-C8 cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aryl and 5-10 membered heteroaryl, wherein the C1-C6 alkyl, C2-C6 alkenyl, C3-C8 cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aryl and 5-10 membered heteroaryl are optionally substituted by 1, 2 or 3 R c Substitution; said R c A substituent selected from the group consisting of H, halogen, hydroxy, amino, nitro, cyano, oxo, carboxyl, C1-C3 alkyl, C2-C4 alkenyl, C1-C3 alkoxy, C1-C3 haloalkyl, C1-C3 haloalkoxy; when R c When it is multiple, the R c Same or different; Optionally, R1 is selected from C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, 3-8 membered heterocycloalkyl, 5-8 membered aryl and 5-8 membered heteroaryl, and the C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, 3-8 membered heterocycloalkyl, 5-8 membered aryl and 5-8 membered heteroaryl are optionally replaced by one R c Substitution; said R c a substituent selected from the group consisting of H, halogen, hydroxy, amino, nitro, cyano, oxo, carboxyl, C1-C3 alkyl, C2-C4 alkenyl, C1-C3 alkoxy, C1-C3 haloalkyl, C1-C3 haloalkoxy; Optionally, R1 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl substituted by C1-C6 alkyl, 4-6 membered heterocycloalkyl, 5-6 membered aryl, 5-6 membered heteroaryl, 5-6 membered aryl substituted by C1-C6 alkyl, 5-6 membered heteroaryl substituted by C1-C6 alkyl; Optionally, R1 is selected from 6. The barbituric acid derivative of formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug according to claim 1, characterized in that: R2 is selected from methyl; Optionally, R3 is selected from H, F, Cl, Br, hydroxy, amino, nitro, cyano, oxo, methyl, methoxy, trifluoromethyl and trifluoromethoxy.

7. The barbituric acid derivative of formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug according to claim 1, characterized in that: The barbituric acid derivatives are selected from the following structures:

8. The method for preparing the barbituric acid derivative of formula II, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug according to claim 2, characterized in that: The method comprises the steps of: Wherein, X1, X2, A, R1, R2, R3 and n are defined as in claim 2.

9. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: the barbituric acid derivative according to any one of claims 1 to 7, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug; and a pharmaceutically acceptable carrier.

10. Use of the barbituric acid derivative according to any one of claims 1 to 7, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug in the preparation of a drug, or use of the pharmaceutical composition according to claim 9 in the preparation of a drug, the use comprising: It is used for treating cardiac diastolic and / or systolic dysfunction, more preferably for treating heart diseases associated with left ventricular hypertrophy and / or diastolic dysfunction, and further preferably for treating hypertrophic cardiomyopathy.

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