Pyrimidinedione derivative as well as preparation method and application thereof

Abstract

The invention provides a pyrimidinedione 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 pyrimidinedione 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 pyrimidinedione derivative and 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 Select C 1 -C 6 Alkylene, carbonyl,

[0013] 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, 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 one or more R a Substitution; said R aA 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 a When it is multiple, the R a Same or different;

[0014] R 2 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 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 3 Selected from H and C 1 -C 6 alkyl;

[0016] m and n are 1, 2 or 3 respectively.

[0017] 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:

[0018]

[0019] Among them, X, R 1 , R 2 , R 3 and n are as defined in the first aspect of the present invention.

[0020] 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.

[0021] In a preferred embodiment of the present invention, the pyrimidinedione derivative has a structure as shown in the following formula I-1: Among them, R 1 , R 2 , R 3 , m and n are defined as described in the first aspect of the present invention.

[0022] In a preferred embodiment of the present invention, the pyrimidinedione derivative has a structure as shown in the following formula I-2: Among them, R 1 , R 2 , R 3 , m and n are defined as described in the first aspect of the present invention.

[0023] In a preferred embodiment of the present invention, the pyrimidinedione derivative has a structure as shown in the following formula I-3: Among them, R 1 , R 2 , R 3 , m and n are defined as described in the first aspect of the present invention.

[0024] In a preferred embodiment of the present invention, the pyrimidinedione derivative has a structure as shown in the following formula I-4: Among them, R 1 , R 2 , R 3 , m and n are defined as described in the first aspect of the present invention.

[0025] In a preferred embodiment of the present invention, R 1 Selected from 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, 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 a Substitution; said R a 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 a When it is multiple, the R a Same or different.

[0026] 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 a Substitution; said R a 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, C1 -C 3 Halogenated alkoxy.

[0027] 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;

[0028] In a preferred embodiment of the present invention, R 1 Selected from

[0029] In a preferred embodiment of the present invention, R 2 is selected from the group consisting of H, F, Cl, Br, hydroxy, amino, nitro, cyano, oxo, methyl, methoxy, trifluoromethyl and trifluoromethoxy.

[0030] In a preferred embodiment of the present invention, R 3 Selected from methyl.

[0031] In a preferred embodiment of the present invention, the pyrimidinedione derivative is selected from any one of the following compounds:

[0032]

[0033] In the third aspect of the present invention, there is provided a method for preparing the pyrimidinedione 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:

[0034]

[0035] Among them, R 1 , R 2 , R 3 and n are defined as described in the second aspect of the present invention.

[0036] In the fourth aspect of the present invention, there is provided another method for preparing the pyrimidinedione 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:

[0037] Among them, R 1 , R 2 , R 3 and n are defined as described in the second aspect of the present invention.

[0038] In the fifth aspect of the present invention, a pharmaceutical composition is provided, comprising: a pyrimidinedione derivative as described in the first aspect or the second aspect of the present invention, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug; and a pharmaceutically acceptable carrier.

[0039] In the sixth aspect of the present invention, there is provided the use of the pyrimidinedione 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 fifth 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.

[0040] 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.

[0041] Terms and Definitions

[0042] 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.

[0043] 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.

[0044] 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.

[0045] 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.

[0046] 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, 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.

[0047] 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.

[0048] 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.

[0049] 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.

[0050] 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.

[0051] 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.

[0052] 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.

[0053] 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.

[0054] 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.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] 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.

[0059] When used alone or as part of another substituent, the term "C 1 -C 6 "Alkylene" refers to a straight-chain divalent hydrocarbon group having 1 to 6 carbon atoms or a branched divalent hydrocarbon group having 3 to 6 carbon atoms, unless otherwise specified, for example, methylene, ethylene, propylene, 1-methylpropylene, butylene, etc.

[0060] 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.

[0061] 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".

[0062] When used alone or as part of another substituent, 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 "C3 -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".

[0063] 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 4 Examples of alkenyl groups include, but are not limited to, vinyl and allyl.

[0064] 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.

[0065] 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.

[0066] As used herein, the term "halogen" alone or as part of another substituent refers to fluorine, chlorine, bromine, iodine; preferably fluorine or chlorine.

[0067] As used herein, the term "cyano" by itself or as part of another substituent means -CN.

[0068] As used herein, the term "amino" by itself or as part of another substituent means -NH 2 .

[0069] As used herein, the term "hydroxy" by itself or as part of another substituent refers to -OH.

[0070] 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.

[0071] 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.

[0072] 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.

[0073] 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.

[0074] 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.

[0075] The term "prevent" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0076] 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.

[0077] 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.

[0078] 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.

[0079] 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.

[0080] Beneficial Effects

[0081] After extensive and in-depth research, the inventors unexpectedly developed a pyrimidine dione 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 pyrimidine dione derivatives provided in the present application will also reduce the rate at which myosin releases ADP and the binding rate with actin when bound to ADP.

[0082] 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

[0083] 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.

[0084] Example 1: Preparation of N-(1-ethyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)benzamide (Compound 1)

[0085] The synthetic route of target compound 1 is as follows:

[0086]

[0087] Step 1: Preparation of Intermediate 1B

[0088] 6-Aminouracil (1A) (5.18 g, 40.7 mmol) was suspended in 25 mL of hexamethyldisilazane (HMDS) and 0.1 mL of sulfuric acid was added. The mixture was heated to 125 °C and refluxed for 3 hours, then concentrated in vacuo. The residue was dissolved in 30 mL of N,N-dimethylformamide (DMF), iodoethane (21.30 g, 136.5 mmol) was added, and stirring was continued at room temperature for 72 h. The reaction was cooled to 0 °C and NaHCO was slowly added. 3 The mixture was stirred at 0°C until no more bubbling occurred. The precipitate was filtered, washed with 10 mL of methanol and 10 mL of water and dried to give Intermediate 1B (4.93 g, yield 78%), which was used without further purification.

[0089] ESI-MS (m / z): 155.2

[0090] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.75 (s; 1H), 6.33 (s; 2H), 4.56 (s; 1H), 3.68-3.75 (q,

[0091] 2H),1.00-1.05(t,3H).

[0092] Step 2: Preparation of N-(1-ethyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)benzamide

[0093] The above intermediate 1B (4.65 g, 30.0 mmol), triethylamine (6.08 g, 60.0 mmol), 4-dimethylaminopyridine (0.61 g, 5.0

[0094] mmol), 30 mL of dichloromethane were added to the reaction flask, and a mixed solution of benzoyl chloride (4.21 g, 30.0 mmol) and 30 mL of dichloromethane was added dropwise at 0°C. The mixture was stirred for 2 hours. After the reaction was completed, the temperature was controlled below 0°C, 50 mL of water was added dropwise to quench the reaction, and the organic layer was separated and eluted with 100 mL of NaHCO 3 The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography (petroleum ether:ethyl acetate = 1:1 as eluent) to obtain compound 1 (4.43 g, yield 57%).

[0095] ESI-MS (m / z): 259.4

[0096] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.75 (s; 1H), 9.95 (s; 1H), 7.76-7.74 (m, 2H), 7.44-7.40 (m,

[0097] 1H),7.34-7.31(m,2H),4.53(s;1H),3.58-3.25(q,2H),0.96-1.03(t,3H).

[0098] Example 2: Preparation of N-(1-n-propyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)benzamide (Compound 2)

[0099] The synthetic route of target compound 2 is as follows:

[0100]

[0101] Step 1: Preparation of intermediate 2B

[0102] 6-Aminouracil (1A) (6.36 g, 50.0 mmol) was suspended in 30 mL of hexamethyldisilazane (HMDS) and 0.1 mL of sulfuric acid was added. The mixture was heated to 125 °C and refluxed for 3 hours, then concentrated in vacuo. The residue was dissolved in 30 mL of N,N-dimethylformamide (DMF), iodopropane (29.75 g, 175.0 mmol) was added, and stirring was continued at room temperature for 60 h. The reaction was cooled to 0 °C and NaHCO was slowly added. 3 The mixture was stirred at 0°C until no more bubbling occurred. The precipitate was filtered, washed with 15 mL of methanol and 15 mL of water and dried to give Intermediate 2B (5.32 g, yield 63%), which was used without further purification.

[0103] ESI-MS (m / z): 169.2

[0104] 1H-NMR(400MHz,DMSO-d6,δ / ppm):10.36(s,1H),6.18(s,2H),4.56(s,1H),3.59-3.65(m,2H),

[0105] 1.42-1.52(m,2H),0.82(t,3H).

[0106] Step 2: Preparation of N-(1-n-propyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)benzamide

[0107] The above intermediate 2B (5.07 g, 30.0 mmol), triethylamine (6.08 g, 60.0 mmol), 4-dimethylaminopyridine (0.61 g, 5.0

[0108] mmol), 30 mL of dichloromethane were added to the reaction flask, and a mixed solution of benzoyl chloride (4.21 g, 30.0 mmol) and 30 mL of dichloromethane was added dropwise at 0°C. The mixture was stirred for 2 hours. After the reaction was completed, the temperature was controlled below 0°C, 50 mL of water was added dropwise to quench the reaction, and the organic layer was separated and eluted with 100 mL of NaHCO 3 The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography (petroleum ether:ethyl acetate = 1:1 as eluent) to obtain compound 2 (4.26 g, yield 52%).

[0109] ESI-MS (m / z): 273.4

[0110] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.71 (s; 1H), 9.98 (s; 1H), 7.73-7.78 (m, 2H), 7.41-7.48 (m,

[0111] 1H),7.32-7.37(m,2H),4.53(s,1H),3.52-3.61(m,2H),1.40-1.53(m,2H),0.85(t,3H,).

[0112] Example 3: Preparation of N-(1-isopropyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)benzamide (Compound 3)

[0113] The synthetic route of target compound 3 is as follows:

[0114]

[0115] Step 1: Preparation of intermediate 3B

[0116] 6-Aminouracil (1A) (3.81 g, 30.0 mmol) was suspended in 20 mL of hexamethyldisilazane (HMDS) and 0.1 mL of sulfuric acid was added. The mixture was heated to 125 °C and refluxed for 5 hours, then concentrated in vacuo. The residue was dissolved in 20 mL of N,N-dimethylformamide (DMF), 2-iodopropane (20.40 g, 120.0 mmol) was added, and stirring was continued at room temperature for 72 h. The reaction was cooled to 0 °C and NaHCO was slowly added. 3The mixture was stirred at 0°C until no more bubbling occurred. The precipitate was filtered, washed with 10 mL of methanol and 10 mL of water and dried to give Intermediate 3B (5.32 g, 47% yield), which was used without further purification.

[0117] ESI-MS (m / z): 169.2

[0118] 1H-NMR(400MHz,DMSO-d6,δ / ppm):10.46(s,1H),6.38(s,2H),4.26(s,1H),4.45(m,1H),1.39(d,

[0119] 6H).

[0120] Step 2: Preparation of N-(1-isopropyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)benzamide

[0121] The above intermediate 3B (5.07 g, 30.0 mmol), triethylamine (6.08 g, 60.0 mmol), 4-dimethylaminopyridine (0.61 g, 5.0

[0122] mmol), 30 mL of dichloromethane were added to the reaction flask, and a mixed solution of benzoyl chloride (4.21 g, 30.0 mmol) and 30 mL of dichloromethane was added dropwise at 0°C. The mixture was stirred for 2 hours. After the reaction was completed, the temperature was controlled below 0°C, 50 mL of water was added dropwise to quench the reaction, and the organic layer was separated and eluted with 100 mL of NaHCO 3 The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography (petroleum ether:ethyl acetate = 1:1 as eluent) to obtain compound 3 (3.77 g, yield 46%).

[0123] ESI-MS (m / z): 273.4

[0124] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.65 (s; 1H), 9.92 (s; 1H), 7.75-7.79 (m, 2H), 7.40-7.50 (m,

[0125] 1H),7.30-7.38(m,2H),4.79(s,1H),4.40(m,1H),1.40(d,6H).

[0126] Example 4: Preparation of N-(1-ethyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)benzenesulfonamide (Compound 8)

[0127] The synthetic route of target compound 8 is as follows:

[0128]

[0129] Intermediate 1B (1.56 g, 10.0 mmol), triethylamine (2.03 g, 20.0 mmol), 4-dimethylaminopyridine (0.12 g, 1.0 mmol) and 15 mL of dichloromethane were added to the reaction flask, and a mixed solution of benzenesulfonyl chloride (1.77 g, 10.0 mmol) and 10 mL of dichloromethane was added dropwise at 0°C. The mixture was stirred for 2 hours. After the reaction was completed, the temperature was controlled below 0°C, 20 mL of water was added dropwise to quench the reaction, and the organic layer was separated and 30 mL of NaHCO 3 The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography (petroleum ether:

[0130] Ethyl acetate = 1:1 as the eluent). Compound 8 (1.51 g, yield 51%) was obtained.

[0131] ESI-MS (m / z): 295.4

[0132] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.65 (s; 1H), 7.86-7.79 (m, 2H), 7.56-7.69 (m, 3H), 7.29 (s;

[0133] 1H),4.23(s;1H),3.88(q,2H),1.13(t,3H).

[0134] Example 9: Preparation of N-(1-propyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)benzenesulfonamide (Compound 9)

[0135] The synthetic route of target compound 9 is as follows:

[0136]

[0137] Intermediate 2B (2.54 g, 15.0 mmol), triethylamine (3.54 g, 35.0 mmol), 4-dimethylaminopyridine (0.18 g, 1.5 mmol) and 25 mL of dichloromethane were added to the reaction flask, and a mixed solution of benzenesulfonyl chloride (2.65 g, 15.0 mmol) and 20 mL of dichloromethane was added dropwise at 0°C. The mixture was stirred for 2 hours. After the reaction was completed, the temperature was kept below 0°C, 20 mL of water was added dropwise to quench the reaction, and the organic layer was separated and 30 mL of NaHCO 3 The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography (petroleum ether:

[0138] Ethyl acetate = 1:1 as the eluent). Compound 9 (2.92 g, yield 63%) was obtained.

[0139] ESI-MS (m / z): 309.4

[0140] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.63 (s; 1H), 7.80 (m, 2H), 7.59 (m, 3H), 7.23 (s; 1H), 4.20

[0141] (m,1H),3.63(m,2H),1.50(m,2H),0.83(t,3H,).

[0142] Example 6: Preparation of N-(1-isopropyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)benzenesulfonamide (Compound 10)

[0143] The synthetic route of target compound 10 is as follows:

[0144]

[0145] Intermediate 3B (1.69 g, 10.0 mmol), triethylamine (2.03 g, 20.0 mmol), 4-dimethylaminopyridine (0.12 g, 1.0 mmol) and 15 mL of dichloromethane were added to a reaction flask, and a mixed solution of benzenesulfonyl chloride (1.77 g, 10.0 mmol) and 10 mL of dichloromethane was added dropwise at 0°C. The mixture was stirred for 2 hours. After the reaction was completed, the temperature was controlled below 0°C, 20 mL of water was added dropwise to quench the reaction, and the organic layer was separated and washed with 30 mL of (1 M)

[0146] NaHCO 3 The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography (petroleum ether:ethyl acetate = 1:1 as eluent) to obtain compound 10 (1.36 g, yield 46%).

[0147] ESI-MS (m / z): 295.4

[0148] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.61 (s; 1H), 7.83 (m, 2H), 7.60 (m, 3H), 7.25 (s; 1H), 4.37

[0149] (m,1H),4.18(s;1H),1.37(d,6H).

[0150] Example 7: Preparation of 6-(benzyl(methyl)amino)-3-ethylpyrimidine-2,4(1H,3H)-dione (Compound 15)

[0151] The synthetic route of target compound 15 is as follows:

[0152]

[0153] Step 1: Preparation of intermediate 15B

[0154] 30 mL of acetic acid was added to the reaction bottle, followed by malonic acid (11.97 g, 115.0 mmol) and N-ethylurea (8.82 g, 100.0 mmol), heated to 70°C, 20 mL of acetic anhydride was added dropwise, and the mixture was heated to 90°C after stirring, stirred for 3 hours until the reaction was completed, cooled to 50°C, and the reaction solution was concentrated under reduced pressure, and then 20 mL of ethanol was added, the mixture was cooled to 0°C, and stirred for 2 hours. Filtered and dried to obtain intermediate 15B (9.53 g, yield 61%).

[0155] ESI-MS (m / z): 156.2

[0156] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 11.30 (s, 1H), 3.70 (q, 2H), 3.59 (s, 2H), 1.06 (t, 3H).

[0157] Step 2: Preparation of intermediate 15C

[0158] The above intermediate 15B (6.23 g, 40.0 mmol) was added with POCl 3 (62.52 g, 40.8 mmol), 1.0 mL of water 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 20 mL of methanol and the obtained solid was recrystallized from ethanol to give Intermediate 15C (2.52 g, yield 36%).

[0159] ESI-MS (m / z): 174.7

[0160] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 12.34 (s, 1H), 5.89 (s, 1H), 3.76 (q, 2H), 1.07 (t, 3H).

[0161] Step 3: Preparation of 6-(benzyl(methyl)amino)-3-ethylpyrimidine-2,4(1H,3H)-dione (Compound 15)

[0162] The above intermediate 15C (1.75 g, 10.0 mmol), N-methylbenzylamine (4.85 g, 40.0 mmol) and 50 mL of 1,4-dioxane were added to a reaction flask, stirred and reacted at 120°C for 12 hours. After the reaction was completed, it was cooled to room temperature and concentrated under reduced pressure. 50 mL of dichloromethane was added and the liquids were separated. The organic layer was washed with 50 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by column chromatography (petroleum ether: ethyl acetate = 1:3 as eluent) to obtain compound 15 (0.98 g, yield 38%).

[0163] ESI-MS (m / z): 259.4

[0164] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.58 (s, 1H), δ7.25–7.37 (m, 5H), 4.86 (2H, s), 4.15 (m,

[0165] 1H),3.90(q,2H),3.07(s;3H),1.13(t,3H).

[0166] Example 8: Preparation of 6-(benzyl(methyl)amino)-3-propylpyrimidine-2,4(1H,3H)-dione (Compound 16)

[0167] The synthetic route of target compound 16 is as follows:

[0168]

[0169] Step 1: Preparation of intermediate 16B

[0170] 30 mL of acetic acid was added to the reaction bottle, followed by malonic acid (11.97 g, 115.0 mmol) and N-propylurea (10.21 g, 100.0 mmol), heated to 70°C, 30 mL of acetic anhydride was added dropwise, and the mixture was stirred and then heated to 100°C, stirred for 5 hours until the reaction was complete, cooled to 50°C, and the reaction solution was concentrated under reduced pressure, and then 50 mL of ethanol was added, the mixture was cooled to 0°C, and stirred for 2 hours. Filtered and dried to obtain intermediate 16B (7.66 g, yield 45%).

[0171] ESI-MS (m / z): 170.2

[0172] 1H-NMR (400MHz, CDCl3, δ / ppm): 8.98 (br, lH), 3.79 (t, 2H), 3.62 (s, 2H), 1.65 (m, 2H), 0.91

[0173] (t,3H).

[0174] Step 2: Preparation of intermediate 16C

[0175] The above intermediate 16B (4.26 g, 25.0 mmol) was added with POCl 3 (4.00 g, 26.0 mmol), 0.4 mL of water 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 20 mL of methanol and the obtained solid was recrystallized from ethanol to give Intermediate 16C (2.03 g, yield 43%).

[0176] ESI-MS (m / z): 188.7

[0177] 1H-NMR (400MHz, CDCl3, δ / ppm): 11.09 (s, 1H), 5.86 (s, 1H), 3.82 (t, 2H), 1.62 (m, 2H), 0.94 (t, 3H).

[0178] Step 3: Preparation of 6-(benzyl(methyl)amino)-3-propylpyrimidine-2,4(1H,3H)-dione (Compound 16)

[0179] The above intermediate 16C (1.89 g, 10.0 mmol), N-methylbenzylamine (4.85 g, 40.0 mmol) and 60 mL of 1,4-dioxane were added to a reaction flask, stirred and reacted at 120°C for 18 hours. After the reaction was completed, it was cooled to room temperature, concentrated under reduced pressure, and 60 mL of dichloromethane was added. The organic layer was washed with 60 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by column chromatography (petroleum ether: ethyl acetate = 1:3 as eluent) to obtain compound 16 (0.87 g, yield 32%).

[0180] ESI-MS (m / z): 273.4

[0181] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.53 (s, 1H), δ7.29–7.35 (m, 5H), 4.84 (2H,s),4.20(m,1H),3.68(m,2H),3.05(s;3H),1.52(m,2H),0.84(t,3H,).

[0182] Example 10: Preparation of 6-(benzyl(methyl)amino)-3-isopropylpyrimidine-2,4(1H,3H)-dione (Compound 17)

[0183] The synthetic route of target compound 17 is as follows:

[0184]

[0185] Step 1: Preparation of intermediate 17B

[0186] 30 mL of acetic acid was added to the reaction bottle, followed by malonic acid (17.17 g, 165.0 mmol) and N-isopropylurea (15.30 g, 131.7 mmol), heated to 70°C, 30 mL of acetic anhydride was added dropwise, and the mixture was heated to 100°C after stirring, stirred for 5 hours until the reaction was complete, cooled to 50°C, and the reaction solution was concentrated under reduced pressure, and then 50 mL of ethanol was added, the mixture was cooled to 0°C, and stirred for 2 hours. Filtered and dried to obtain intermediate 17B (11.66 g, yield 52%).

[0187] ESI-MS (m / z): 170.2

[0188] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 11.19 (s, 1H), 4.83 (m, 1H), 3.58 (s, 2H), 1.32 (d, 6H).

[0189] Step 2: Preparation of intermediate 17C

[0190] The above intermediate 17B (8.51 g, 50.0 mmol) was added with POCl 3 (8.00 g, 52.0 mmol), 0.6 mL of water 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 40 mL of methanol and the obtained solid was recrystallized from ethanol to give Intermediate 17C (3.49 g, yield 37%).

[0191] ESI-MS (m / z): 188.7

[0192] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 12.22 (s, 1H), 5.88 (s, 1H), 4.95 (m, 1H), 1.34 (d, 6H).

[0193] Step 3: Preparation of 6-(benzyl(methyl)amino)-3-isopropylpyrimidine-2,4(1H,3H)-dione (Compound 17)

[0194] The above intermediate 17C (2.83 g, 15.0 mmol), N-methylbenzylamine (9.00 g, 75.0 mmol) and 80 mL of 1,4-dioxane were added to a reaction flask, stirred and reacted at 120°C for 15 hours. After the reaction was completed, it was cooled to room temperature and concentrated under reduced pressure. 100 mL of dichloromethane was added and the liquids were separated. The organic layer was washed with 100 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by column chromatography (petroleum ether: ethyl acetate = 1:3 as eluent) to obtain compound 17 (1.15 g, yield 28%).

[0195] ESI-MS (m / z): 273.4

[0196] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.60 (s, 1H), δ7.25–7.33 (m, 5H), 4.76 (2H, s), 4.40 (m, 1H),

[0197] 4.18(s;1H),3.05(s;3H),1.37(d,6H).

[0198] Example 11: Preparation of N-(1-ethyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)-N-methylbenzamide (Compound 21)

[0199] The synthetic route of target compound 21 is as follows:

[0200]

[0201] Step 1: Preparation of intermediate 21D

[0202] In a reaction flask, intermediate 15C (4.36 g, 25.0 mmol), methylamine (40%, W / W) aqueous solution (2.52 g, 32.50 mmol), triethylamine (5.06 g, 50.00 mmol) and 40 mL of ethanol were added, and the mixture was refluxed for 12 hours. After the reaction was completed, the mixture was cooled to room temperature, and the reaction solution was concentrated under reduced pressure. Then, 20 mL of ether was added, and the mixture was cooled to 0°C and stirred for 2 hours. After filtration and drying, intermediate 21D (2.24 g, yield 53%) was obtained.

[0203] ESI-MS (m / z): 169.3.

[0204] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.52 (s, 1H), 8.76 (m, 1H), 4.32 (s, 1H), 3.79 (q, 2H), 1.16 (t, 3H).

[0205] Step 2: Preparation of N-(1-ethyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)-N-methylbenzamide (Compound 21)

[0206] The intermediate 21D (5.10 g, 30.0 mmol), triethylamine (6.10 g, 60.0 mmol), 4-dimethylaminopyridine (0.61 g, 5.0 mmol) and 100 mL of dichloromethane were added to a reaction flask, and a mixed solution of benzoyl chloride (4.21 g, 30.0 mmol) and 50 mL of dichloromethane was added dropwise at 0°C. The reaction was stirred for 5 hours. After the reaction was completed, the temperature was controlled below 0°C, 80 mL of water was added dropwise to quench, and the organic layer was separated and 100 mL of NaHCO 3 The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography (petroleum ether:ethyl acetate=1:2 as eluent) to obtain compound 21 (3.03 g, yield 37%).

[0207] ESI-MS (m / z): 273.4

[0208] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.63 (s; 1H), 7.96 (m, 2H), 7.54-7.65 (m, 3H), 4.83 (s; 1H), 3.84

[0209] (q, 2H), 3.58 (s, 3H), 1.20 (t, 3H).

[0210] Example 12: Preparation of N-(1-propyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)-N-methylbenzamide (Compound 22)

[0211] The synthetic route of target compound 22 is as follows:

[0212]

[0213] Step 1: Preparation of intermediate 22D

[0214] In a reaction flask, intermediate 15C (2.36 g, 12.5 mmol), methylamine (40%, W / W) aqueous solution (1.26 g, 16.25 mmol), triethylamine (2.53 g, 25.00 mmol) and 25 mL of ethanol were added, and the reaction was refluxed for 12 hours. After the reaction was completed, the mixture was cooled to room temperature, and the reaction solution was concentrated under reduced pressure. Then 15 mL of ether was added, and the mixture was cooled to 0°C and stirred for 2 hours. Filtered and dried to obtain intermediate 22D (0.78 g, yield 34%).

[0215] ESI-MS (m / z): 183.3.

[0216] 1H-NMR(400MHz,DMSO-d6,δ / ppm):10.56(s,1H),8.70(m,1H),4.26(s,1H),3.65(m,2H),1.48(m,

[0217] 2H),0.85(t,3H).

[0218] Step 2: Preparation of N-(1-propyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)-N-methylbenzamide (Compound 22)

[0219] The intermediate 22D (1.10 g, 6.0 mmol), triethylamine (1.22 g, 12.0 mmol), 4-dimethylaminopyridine (0.12 g, 1.0 mmol) and 30 mL of dichloromethane were added to a reaction flask, and a mixed solution of benzoyl chloride (0.84 g, 6.0 mmol) and 30 mL of dichloromethane was added dropwise at 0°C. The mixture was stirred for 4 hours. After the reaction was completed, the temperature was controlled below 0°C, 20 mL of water was added dropwise to quench the reaction, and the organic layer was separated and 30 mL of NaHCO 3 The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography (petroleum ether:ethyl acetate=1:2 as eluent) to obtain compound 22 (0.55 g, yield 32%).

[0220] ESI-MS (m / z): 287.4

[0221] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.59 (s; 1H), 7.54-7.95 (m, 5H), 4.89 (s; 1H), 3.65 (m, 2H), 3.58

[0222] (s, 3H), 1.48 (m, 2H), 0.85 (t, 3H).

[0223] Example 13: Preparation of N-(1-ethyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)-N-methylbenzenesulfonamide (Compound 24)

[0224] The synthetic route of target compound 24 is as follows:

[0225]

[0226] The intermediate 21D (1.70 g, 10.0 mmol), triethylamine (2.05 g, 20.0 mmol), 4-dimethylaminopyridine (0.61 g, 5.0 mmol) and 300 mL of dichloromethane were added to a reaction flask, and a mixed solution of benzoyl chloride (1.40 g, 10.0 mmol) and 20 mL of dichloromethane was added dropwise at 0°C. The mixture was stirred for 6 hours. After the reaction was completed, the temperature was controlled below 0°C, 30 mL of water was added dropwise to quench the reaction, and the organic layer was separated and 30 mL of NaHCO 3 The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography (petroleum ether:ethyl acetate=1:2 as eluent) to obtain compound 24 (0.83 g, yield 27%).

[0227] ESI-MS (m / z): 309.4

[0228] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.55 (s; 1H), 7.83 (m, 2H), 7.56-7.69 (m, 3H), 4.29 (s; 1H), 3.90

[0229] (q;2H),2.68(s,3H),1.23(t,3H).

[0230] Example 14: Preparation of N-(1-propyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)-N-methylbenzenesulfonamide (Compound 25)

[0231] The synthetic route of target compound 25 is as follows:

[0232]

[0233] The intermediate 22D (0.55 g, 5.0 mmol), triethylamine (0.61 g, 6.0 mmol), 4-dimethylaminopyridine (0.12 g, 1.0 mmol) and 20 mL of dichloromethane were added to a reaction flask, and a mixed solution of benzoyl chloride (0.42 g, 3.0 mmol) and 20 mL of dichloromethane was added dropwise at 0°C. The mixture was stirred for 5 hours. After the reaction was completed, the temperature was controlled below 0°C, 20 mL of water was added dropwise to quench the reaction, and the organic layer was separated and 30 mL of NaHCO 3 The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography (petroleum ether:ethyl acetate=1:2 as eluent) to obtain compound 25 (0.38 g, yield 24%).

[0234] ESI-MS (m / z): 323.4

[0235] 1H-NMR (400MHz, DMSO-d6, δ / ppm): 10.68 (s; 1H), 7.79 (m, 2H), 7.60 (m, 3H), 4. 25(s;1H),4.20(m,1H),3.68(t,2H),3.65(s,3H),1.54(m,2H),0.88(t,3H,).

[0236] The following target compounds were prepared similarly to the synthetic methods of the above examples.

[0237]

[0238]

[0239]

[0240] Test Example 1: Determination of myosin inhibitory activity

[0241] 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 .

[0242] 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, M200PRO-grating multifunctional microplate detector, 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. Then the normalized ratio is plotted as a function of drug concentration, and the data is fitted into a curve using Graph PadPrism. The midpoint of the curve is IC50, which is the drug concentration when 50% of the total response is inhibited.

[0243] The results of the tested compounds are shown in Table 1. The tested compounds were prepared according to the synthesis method described herein.

[0244] Table 1 Myosin inhibitory activity of the compounds of the present invention

[0245]

[0246] The experimental results show that the compounds disclosed herein have an inhibitory effect on myosin ATPase.

[0247] Test Example 2: Contractility measurement of adult rat cardiomyocytes

[0248] Adult male rats were anesthetized and the heart was rapidly removed, flushed and the ascending aorta cannulated. Continuous retrograde perfusion was initiated on the heart at a perfusion pressure of 60 cmH2O. The heart was first perfused with a nominally Ca2+-free modified Krebs solution having the following composition: 113 mM NaCl, 4.7 mM KCl, 0.6 mM KH 2 PO 4 , 0.6mMNa 2 HPO 4 、1.2mMMgS0 4 , 12 mM NaHCO 3 , 10 mM 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 render 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 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 treatment with standards (>150% of basal) and isoproterenol (ISO; >250% of basal). In addition, only cells with a basal contraction between 3% and 8% were used in subsequent experiments with compounds.

[0249] The myocyte aliquots in Tyrode's buffer were placed in a perfusion chamber equipped with a heating stage. The myocytes were attached, the chamber was heated to 37°C, and the cells were perfused with 37°C Tyrode's buffer. The myocytes were stimulated with a 1Hz (20% above threshold) electric field using a platinum electrode. Only cells with clear stripes and resting before pacing were used for contractility experiments. To determine the basal contraction rate, myocytes were imaged via a 40x objective. Using a variable frame rate (60Hz-240Hz) charge-coupled device camera, the images were digitized and displayed on a computer screen (IonOptix Milton) at a sampling rate of 240Hz. Once the cell contraction was stable over time, the test compound (0.01μM-15μM) was perfused into the chamber of the myocyte for 5 minutes. Edge detection was then used to record the contraction rate of the myocytes as well as the contraction and relaxation speeds. The data were continuously recorded using IonOptix software. Contractility data were analyzed using Ionwizard software (IonOptix), and for each cell, 10-20 contractility transients were averaged and compared to baseline (no compound) conditions and conditions treated with compound. Compounds were measured by their effect on fractional shortening (FS), which is the ratio of the maximum length of the cell during contraction divided by the baseline cell length and normalized to 100% for untreated cells.

[0250] Table 2 Inhibition of cardiomyocyte contraction by selected compounds

[0251]

[0252]

[0253] + 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.

[0254] The experimental results showed that at the same molar concentration, the inhibitory effects of compounds 3 and 10 were equivalent to that of MYK-461.

[0255] Test Example 3: Pharmacokinetics of the compound in rats

[0256] Nine male SD rats were divided into two groups, 3 rats in each group, and were given test compounds 3 and 10 by intravenous injection and oral gavage 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 concentrations of compounds 3 and 10 in plasma samples were detected by LC-MS / MS method, and the software Calculate pharmacokinetic parameters.

[0257] Table 3: Pharmacokinetic parameters of compounds 3 and 10 after oral administration to rats

[0258]

[0259] The experimental results show that the pharmacokinetic absorption of compounds 3 and 10 in SD rats is good. In addition, the T of compounds 3 and 10 in the present invention is better than that of compound MYK-461. 1 / 2 The 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 compounds 3 and 10 disclosed in the present invention have obvious pharmacokinetic advantages over the compound MYK-461.

[0260] 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 pyrimidinedione derivative of formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug: in, X is selected from C1-C6 alkylene, carbonyl, and 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 a Substitution; said R a 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 a When it is multiple, the R a Same or different; R2 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 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; R3 is selected from H and C1-C6 alkyl; m and n are 1, 2 or 3 respectively.

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

3. The pyrimidinedione derivative of formula I according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that: The pyrimidinedione derivative is any of the following schemes: Scheme 1: The pyrimidinedione derivative has a structure as shown in the following formula I-1: Wherein, R1, R2, R3, m and n are defined as in claim 1; Scheme 2: The pyrimidinedione derivative has a structure as shown in the following formula I-2: Wherein, R1, R2, R3, m and n are defined as in claim 1; Scheme 3: The pyrimidinedione derivative has a structure as shown in the following formula I-3: Wherein, R1, R2, R3, m and n are defined as in claim 1; Scheme 4: The pyrimidinedione derivative has a structure as shown in the following formula I-4: Wherein, R1, R2, R3, m and n are as defined in claim 1.

4. The pyrimidinedione derivative of formula I according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, 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 a Substitution; said R a 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 a 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 a Substitution; said R a 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 5. The pyrimidinedione derivative of formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug according to claim 1, characterized in that: R3 is selected from methyl; Optionally, R2 is selected from H, F, Cl, Br, hydroxy, amino, nitro, cyano, oxo, methyl, methoxy, trifluoromethyl and trifluoromethoxy.

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

7. A method for preparing the pyrimidinedione derivative of formula II, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug according to claim 2, characterized in that: The method is any of the following: Option A: Wherein, R1, R2, R3 and n are defined as in claim 2; Option B: Wherein, R1, R2, R3 and n are defined as in claim 2.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: the pyrimidinedione derivative according to any one of claims 1 to 6, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug; and a pharmaceutically acceptable carrier.

9. Use of the pyrimidinedione derivative according to any one of claims 1 to 6, 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 8 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 ventricular hypertrophy and / or diastolic dysfunction, and further preferably for treating hypertrophic cardiomyopathy.

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