A deuterated pyrazole sulfonylmethyl-piperidinyl isoxazole urea compound and its use

Abstract

The present invention discloses a deuterated pyrazolesulfonylmethyl-piperidinylisoxazole urea compound and its use, and discloses a compound shown in formula I, or a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate thereof, a pharmaceutical composition and use. The compound shown in formula I provided by the present invention has a good therapeutic effect on primary dilated cardiomyopathy as a myocardial actin activator. #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of innovative pharmaceutical chemistry and relates to a deuterated pyrazole compound, a pharmaceutical composition and uses. Background Art

[0002] Dilated cardiomyopathy (DCM) includes a group of myocardial diseases that cause left ventricular dilation and systolic dysfunction (abnormal contraction), accounting for 90% of cardiomyopathy and is one of the main causes of heart failure. DCM can be subdivided into ischemic (attributed to coronary artery disease) or non-ischemic (primary myocardial disease). Heart failure (HF) is the terminal stage of the development of various cardiovascular diseases and seriously threatens human life and health. The high incidence of cardiovascular diseases has become one of the main causes of mortality worldwide. Therefore, the development of anti-heart failure drugs is very important. Decreased / impeded myocardial contraction is the main sign of heart failure. In order to maintain a stable cardiac output, the body compensates by increasing sympathetic nerve tension and activating the neuroendocrine system to increase myocardial contractility, but this often accelerates the progression of heart failure. At present, anti-heart failure drugs mainly include neuroendocrine blockers and positive inotropic drugs. The characteristics of the action of neuroendocrine blockers are to improve neuroendocrine abnormalities, block sympathetic excitation, reduce ventricular remodeling, reduce peripheral vascular resistance, reduce the afterload of the heart and further block the vicious cycle. However, this type of drug cannot fundamentally improve the condition of insufficient myocardial contractility and has certain limitations. The characteristic of positive inotropic drugs is to strengthen myocardial contractility and increase cardiac output. Traditional positive inotropic drugs include β-adrenergic receptor agonists, cardiac glycosides, phosphodiesterase inhibitors and calcium sensitizers.

[0003] β-adrenergic receptor agonists increase the concentration of calcium ions in myocardial cells through the second messenger cAMP pathway, thereby enhancing myocardial contractility. Cardiac glycosides can inhibit the Na + / K + -ATPase activity, making Na + -K + Exchange is reduced due to Na + Cannot be actively pumped out of the membrane, making the Na + Increase, thus stimulating Na + -Ca 2+ exchange system, promoting Na + Efflux, Ca 2+ Influx, intramembrane Ca 2+Increase, produce positive inotropic effect. However, these drugs can cause side effects such as increased heart rate, decreased blood pressure, increased myocardial oxygen consumption and arrhythmia, and cannot improve prognosis or reduce mortality. Calcium sensitizers can directly increase the sensitivity of myocardial contractile proteins to calcium ions after entering myocardial cells, and play a positive inotropic role without affecting myocardial oxygen consumption. However, calcium sensitizers usually have phosphodiesterase inhibitory activity, and are prone to side effects such as headache and hypotension in clinical practice. Therefore, the development of anti-heart failure drugs with positive inotropic effects and few side effects is a key issue to be solved in drug research and development. In recent years, the development of cardiac myosin agonists has become a research hotspot for positive inotropic drugs. Cardiac myosin is a downstream target for regulating myocardial contraction. Activation of cardiac myosin can effectively increase myocardial contractility without affecting the intracellular calcium ion concentration. New positive inotropic drugs targeting cardiac myosin are expected to become new treatments for heart failure.

[0004] MYK-491 (Danicamtiv) is a selective cardiac myosin agonist that increases calcium sensitivity of ventricular and atrial myocardial fibers, increases myosin ATPase activity, thereby increasing the molecular force-generating units of the sarcomere, prolonging contractions while maintaining actin-myosin separation, allowing relaxation without sustained contraction, and does not affect Ca 2+ Steady state. Currently in Phase II clinical research for the treatment of primary dilated cardiomyopathy.

[0005] Deuterated drugs refer to drugs in which some hydrogen atoms in the drug molecule are replaced with deuterium. Since the shape and volume of deuterium in the drug molecule are similar to those of hydrogen, deuterated drugs generally retain the biological activity and selectivity of the original drug. Since the CD bond is more stable than the CH bond, the CD bond of deuterated drugs is less likely to break during the chemical reaction, and its half-life will be extended. Since 2000, the deuterated strategy has been widely used in drug research.

[0006] Summary of the invention

[0007] The present invention provides a compound as shown in Formula I or a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate thereof, the structure of which is as follows:

[0008]

[0009] wherein R1, R2, R3 or R4 are independently selected from hydrogen or deuterium,

[0010] At the same time, at least one of R1, R2, R3 or R4 is deuterium.

[0011] In some embodiments, the compound is represented by any of the following structural formulas:

[0012]

[0013] The present invention provides a use of a compound as shown in formula I or a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate thereof in the preparation of a cardiac myosin agonist.

[0014] The present invention provides a use of a compound shown in I or a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate thereof in the preparation of a medicament for preventing and / or treating dilated cardiomyopathy, or a cardiac disease with pathophysiological characteristics of dilated cardiomyopathy, or a disease with contractile dysfunction or reduced contractile reserve.

[0015] The present invention provides a use of a compound shown in I or a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate thereof in the preparation of a medicament for preventing and / or treating diseases or conditions such as systolic dysfunction, diastolic dysfunction, HFrEF, HFpEF, chronic heart failure and acute heart failure.

[0016] The present invention provides use of a compound as shown in I or a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate thereof in the preparation of a medicament for preventing and / or treating a disease or condition characterized by left ventricular systolic dysfunction or symptoms or reduced exercise capacity due to systolic dysfunction.

[0017] The present invention provides a pharmaceutical composition, which contains a compound as shown in Formula I, or a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate thereof, and a pharmaceutically acceptable carrier or excipient.

[0018] In the pharmaceutical composition, the compound as shown in Formula I, or its pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate is used in a therapeutically effective amount.

[0019] The present invention provides use of a pharmaceutical composition in preparing a cardiac myosin agonist.

[0020] The present invention provides a use of a pharmaceutical composition in preparing a drug for preventing and / or treating dilated cardiomyopathy, or a heart disease with pathological and physiological characteristics of dilated cardiomyopathy, or a disease with contractile dysfunction or reduced contractile reserve.

[0021] The present invention provides a use of a pharmaceutical composition in preparing a medicament for preventing and / or treating diseases or conditions of systolic dysfunction, diastolic dysfunction, HFrEF, HFpEF, chronic heart failure and acute heart failure.

[0022] The present invention provides a use of a pharmaceutical composition in the preparation of a medicament for preventing and / or treating a disease or condition characterized by left ventricular systolic dysfunction or symptoms or reduced exercise capacity due to systolic dysfunction.

[0023] The pharmaceutical excipients can be those widely used in the field of drug production. Excipients are mainly used to provide a safe, stable and functional pharmaceutical composition, and can also provide methods to dissolve the active ingredient at a desired rate after the subject receives the administration, or promote the effective absorption of the active ingredient after the subject receives the composition. The pharmaceutical excipients can be inert fillers, or provide a certain function, such as stabilizing the overall pH value of the composition or preventing the degradation of the active ingredient of the composition. The pharmaceutical excipients can include one or more of the following excipients: adhesives, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesive agents, glidants, wetting agents, gelling agents, absorption delay agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents and sweeteners.

[0024] The pharmaceutical composition of the present invention can be prepared according to the disclosed content using any method known to those skilled in the art, such as conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding or lyophilizing processes.

[0025] The pharmaceutical composition of the present invention can be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid preparations), inhalation, ocular, rectal, topical or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intraarterial, intramuscular) administration. The pharmaceutical composition of the present invention can also be a controlled release or delayed release dosage form (e.g., liposomes or microspheres). Examples of solid oral preparations include, but are not limited to, powders, capsules, caplets, soft capsules, and tablets. Examples of liquid preparations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs, and solutions. Examples of topical preparations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum preparations. Examples of preparations for parenteral administration include, but are not limited to, solutions for injection, dry preparations that can be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection. Examples of other suitable formulations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic preparations; aerosols such as nasal sprays or inhalers; liquid dosage forms suitable for parenteral administration; suppositories and lozenges.

[0026] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared from compounds having specific substituents discovered by the present invention with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the free form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the free form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid (forming carbonate or bicarbonate), phosphoric acid (forming phosphate, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid (forming sulfate or bisulfate), hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid. Acids such as tartaric acid and methanesulfonic acid; organic acid salts also include salts of amino acids (such as arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups, and can be converted into any base or acid addition salt. Preferably, the salt is contacted with a base or acid in a conventional manner, and the parent compound is separated, thereby regenerating the free form of the compound. The free form of the compound differs from its various salt forms in certain physical properties, such as different solubility in polar solvents.

[0027] The "pharmaceutically acceptable salts" of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. In general, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of an appropriate base or acid in water or an organic solvent or a mixture of the two. In general, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred.

[0028] The term "isomers" refers to compounds that have the same chemical formula but different arrangements of the atoms.

[0029] The term "metabolite" refers to a pharmaceutically active product produced by the in vivo metabolism of a compound of Formula I or a salt thereof. Such a product may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, glucuronidation, enzymatic cleavage, etc. of the administered compound. Therefore, the present invention includes metabolites of the compounds of the present invention, including compounds produced by a process of contacting a compound of the present invention with a mammal for a period of time sufficient to obtain a metabolite thereof.

[0030] Metabolites are typically identified by preparing radiolabeled isotopes of the compounds of the invention, administering them parenterally to animals, such as rats, mice, guinea pigs, monkeys, or humans, at detectable doses (e.g., greater than about 0.5 mg / kg), allowing sufficient time for metabolism to occur (typically about 30 seconds to 30 hours), and isolating the conversion products from urine, blood, or other biological samples. These products are easily isolated because they are labeled (others are isolated by using antibodies that can bind to antigenic epitopes present in the metabolites). The metabolite structure is determined in a conventional manner, for example, by MS, LC / MS or NMR analysis. Generally, the analysis of metabolites is performed in the same manner as conventional drug metabolism studies known to those skilled in the art. As long as the metabolite products are not otherwise not found in vivo, they can be used in assays for therapeutic dosing of the compounds of the invention. The compounds of the invention may contain non-natural proportions of atomic isotopes on one or more of the atoms that make up the compound. For example, compounds may be labeled with radioisotopes, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C) All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

[0031] In addition to the form of salts, the compounds provided by the present invention also exist in the form of prodrugs. The prodrugs of the compounds described herein are easily chemically changed under physiological conditions to be converted into the compounds of the present invention. Any compound that can be converted in vivo to provide a bioactive substance (i.e., a compound shown in Formula I) is a prodrug within the scope and spirit of the present invention. For example, a carboxyl-containing compound can form a physiologically hydrolyzable ester, which acts as a prodrug by being hydrolyzed in vivo to obtain the compound shown in Formula I itself. The prodrug is preferably administered orally, because hydrolysis occurs mainly under the influence of digestive enzymes in many cases. When the ester itself is active or hydrolysis occurs in the blood, parenteral administration can be used.

[0032] The positive and progressive effects of the present invention are:

[0033] (1) The compounds of the present invention significantly increase the calcium sensitivity of myocardial fibers and increase the activity of myocardial myosin ATPase.

[0034] (2) The compounds of the present invention have excellent pharmacokinetic properties, significantly improved oral bioavailability, and prolonged half-life, which can reduce the single dose, extend the dosing interval, and reduce the number of dosing times.

[0035] (3) The compounds of the present invention have a good therapeutic effect on primary dilated cardiomyopathy. DETAILED DESCRIPTION

[0036] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0037] Example 1: Synthesis of Compound I-1

[0038]

[0039] Step 1: Synthesis of compound b

[0040] The raw material a (2.9 g, 30 mmol) was dissolved in acetone (100 mL), potassium carbonate (6.2 g, 45 mmol) and deuterated iodomethane (6.4 g, 45 mmol) were added to the above solution, the temperature was raised to 40°C, and the reaction was stirred for 2 h. After the reaction was completed, the filtrate was filtered, concentrated, ethanol was slurried, filtered, the filter cake was collected, and vacuum dried to obtain compound b (1.9 g, 55%). MS (ESI, m / z): 114 (M + +1).

[0041] Step 2: Synthesis of compound c

[0042] NBS (2.4 g, 13.2 mmol) was added to a DMF (10 mL) solution of compound b (1.5 g, 13.2 mmol), and the reaction solution was heated to 50°C and stirred for 2 h. After the reaction was complete, ice water was added to quench the reaction, cooled to -20°C and allowed to stand overnight, a large amount of precipitation was precipitated, filtered, the filter cake was collected, and vacuum dried to obtain compound c (2.0 g, 80%). MS (ESI, m / z): 191 (M + +1).

[0043] Step 3: Synthesis of compound d

[0044] Pd2(dab)3CCl3 (1 g, 0.98 mmol), potassium carbonate (6.9 g, 50 mmol), xantphos (1.2 g, 1.97 mmol), potassium iodide (332 mg, 2.00 mol) and compound h (6.5 g, 24 mmol) were added to a solution of compound c (3.8 g, 20 mmol) in 1,4-dioxane (40 mL). The reaction solution was heated to 80°C and a catalytic amount of methanol was added dropwise. The reaction mixture was heated to 85°C and stirred for 12 h. The reaction solution was cooled to 0°C, the solid was filtered off, the filtrate was concentrated, and the compound d (4.1 g, 60%) was obtained by column chromatography separation and purification. MS (ESI, m / z): 343 (M + +1).

[0045] Step 4: Synthesis of compound e

[0046] Compound d (3.43 g, 10 mmol) was dissolved in DCM (30 mL). DAST (6.5 g, 40 mmol) was added dropwise to the above solution at 0°C. The reaction solution was transferred to room temperature and reacted overnight. After the reaction was completed, the reaction solution was cooled to 0°C, quenched with saturated sodium bicarbonate solution, extracted with ethyl acetate (20 mL × 3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and purified by column chromatography to obtain compound e (2.4 g, 66%). MS (ESI, m / z): 365 (M + +1).

[0047] Step 5: Synthesis of compound f

[0048] To compound e (3.65 g, 10 mmol) and ruthenium trichloride (64 mg, 0.32 mmol) in THF / water (v / v=1:1, 50 mL) was added a solution of sodium periodate (8.6 g, 40 mmol) in water (4 mL). The reaction solution was stirred at room temperature for 1 h. After the reaction was complete, water was added to quench the reaction, and ethyl acetate was extracted (20 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and purified by column chromatography to obtain compound f (1.8 g, 46%). MS (ESI, m / z): 397 (M + +1).

[0049] Step 6: Synthesis of compound g

[0050] At -78°C, LDA (2.0M, 5mL, 10mmol) was added dropwise to a solution of compound f (1.98g, 5mmol) in anhydrous THF (30mL), followed by a solution of NFSI (2.1g, 5mmol) in THF. The reaction solution was stirred at -78°C for 30min, and a saturated ammonium chloride solution was added to quench the reaction. The mixture was extracted with ethyl acetate (10mL×2), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound g (1.0g, 50%). MS (ESI, m / z): 415 (M + +1).

[0051] Step 7: Synthesis of Compound 1

[0052] At -78°C, a THF solution of potassium tert-butoxide (1.0 M, 10 mL, 10 mmol) was added to a solution of compound g (2.10 g, 5 mmol) in anhydrous THF (30 mL). The reaction mixture was stirred for 5 min, and then iodomethane was added dropwise. After the reaction mixture was stirred for 15 min, a saturated ammonium chloride solution was added to quench the reaction, and the mixture was extracted with ethyl acetate (10 mL×2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, separated and purified by column chromatography, and then separated by a chiral column to obtain compound 1 (749 mg, 35%). MS (ESI, m / z): 429 (M + +1).

[0053] Step 8: Synthesis of Compound I-1

[0054] The raw material 1 (181 mg, 0.5 mmol) was dissolved in acetonitrile (2 mL), and triethylamine (0.21 mL, 1.5 mmol) and compound 2 (112 mg, 0.55 mmol) were added to the above solution, and the reaction solution was heated to 70°C and stirred overnight. After cooling to room temperature, water was added to quench the reaction, and ethyl acetate was extracted (10 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated and purified by column chromatography to obtain compound I-1 (109 mg, 50%). 1 H NMR (500MHz, DMSO-d6) δ9.82 (s, 1H), 8.90 (s, 1H), 8.85 (d, J = 1.8Hz, 1H), 7.44 (s, 1H), 6.72 (d, J = 1.8Hz, 1H), 3. 96 (s, 2H), 3.87 (ddd, J=11.9, 7.9, 5.8Hz, 2H), 2.04 (s, 2H), 1.99 (s, 2H), 1.93-1.77 (m, 4H). MS (ESI, m / z): 439 (M + +1).

[0055] Example 2: Synthesis of Compound I-2

[0056]

[0057] Step 1: Synthesis of compound 4

[0058] Potassium hydroxide (105.5 mg, 1.88 mmol) and iodine (239 mg, 0.94 mmol) were added to a solution of compound 3 (200 mg, 0.47 mmol) in N, N-dimethylformamide (15 mL), and the mixture was reacted at room temperature for 3 hours. The reaction was complete after monitoring by TLC. A saturated sodium sulfite solution was added to quench the reaction. The aqueous phase was extracted with ethyl acetate (10 mL × 2), washed with water (20 mL × 2), washed with saturated salt (20 mL), dried over anhydrous sodium sulfate, and separated and purified by column chromatography to obtain iodinated compound 2 (111 mg, 43%). MS (ESI, m / z): 552 (M + +1).

[0059] Step 2: Synthesis of compound 5

[0060] Sodium acetate (49 mg, 0.36 mmol) was added to a deuterated acetic acid solution (4 mL) of compound 4 (99 mg, 0.18 mmol), and the mixture was added dropwise over 2 hours. The mixture was reacted at room temperature for 24 hours. The reaction was complete after TLC detection. The mixture was concentrated under reduced pressure and purified by column chromatography to obtain compound 5 (38 mg, 50%). MS (ESI, m / z): 427 (M++1).

[0061] Step 3: Synthesis of compound 6

[0062] The raw material 5 (213 mg, 0.5 mmol) was dissolved in EA (2 mL), and 4MEA / HCl solution (1 mL) was added to the above solution, and the mixture was stirred at room temperature for 3 h. After the reaction was complete, the mixture was filtered and vacuum dried to obtain compound 6, which was directly used for the next step reaction.

[0063] Step 4: Synthesis of Compound I-2

[0064] The raw material 6 (181 mg, 0.5 mmol) was dissolved in acetonitrile (2 mL), and triethylamine (0.21 mL, 1.5 mmol) and compound 2 (112 mg, 0.55 mmol) were added to the above solution, and the reaction solution was heated to 70°C and stirred overnight. After cooling to room temperature, water was added to quench the reaction, and ethyl acetate was extracted (10 mL x 3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated and purified by column chromatography to obtain compound I-2 (109 mg, 50%). 1H NMR (500MHz, DMSO-d6) δ9.82 (s, 1H), 8.85 (d, J = 1.8Hz, 1H), 7.44 (s, 1H), 6.72 (d, J = 1.8Hz, 1H), 3.96 (s, 2H), 3.87 (ddd, J = 11.9, 7.9, 5.8H z, 2H), 2.70 (ddd, J=11.9, 7.9, 5.7Hz, 2H), 2.38 (dp, J=25.1, 5.7Hz, 1H), 2.04 (s, 2H), 1.99 (s, 2H), 1.93-1.77 (m, 4H). MS (ESI, m / z): 437 (M + +1).

[0065] Example 3: Synthesis of Compound I-3

[0066]

[0067] The synthesis method is as shown in Example 2, and it is only necessary to replace the raw material compound 3 with compound 1. 1 H NMR (500MHz, DMSO-d6) δ9.82 (s, 1H), 8.85 (d, J = 1.8Hz, 1H), 7.44 (s, 1H), 6.72 (d, J = 1.8Hz, 1H), 3.96 (s, 2 H), 3.87 (ddd, J=11.9, 7.9, 5.8Hz, 2H), 2.04 (s, 2H), 1.99 (s, 2H), 1.93-1.77 (m, 4H). MS (ESI, m / z): 440 (M + +1).

[0068] Example 4: Myosin Activation Assay

[0069] The ADP released from cardiac myosin is measured by a biochemical method that combines an enzyme coupling system consisting of pyruvate kinase and lactate dehydrogenase. The ability of the test compound to activate the enzymatic activity of bovine cardiac myosin is evaluated by the decrease in the absorbance of NADH at 340nm. Experimental principle: Pyruvate kinase converts ADP into ATP by converting phosphoenolpyruvate into pyruvate. Lactate dehydrogenase converts pyruvate into lactate by converting NADH into NAD. The source of cardiac myosin is bovine heart in the form of myofibrils without sarcolemma. For specific experimental operations, refer to the experimental method disclosed in WO2016 / 118774. AC 20 The concentration of compound corresponding to the total response greater than 20% (μM).

[0070] Table 1 Myosin activation activity of the tested compounds

[0071] Compound Name <![CDATA[AC 20 (μM)]]> I-1 0.28 I-2 0.29 I-3 0.36 MYK-491 0.72

[0072] As shown in Table 1, compounds I-1 to I-3 can significantly increase the activity of myosin and are superior to the positive control compound MYK-491.

[0073] Example 5: Cardiomyocyte contractility assay

[0074] The contractility of adult rat ventricular myocytes was determined by edge detection of the IonOptix contractility system. The specific experimental operation refers to the experimental method disclosed in WO2016 / 118774. The myocytes were aliquoted in Tyrode buffer (137mM NaCl, 3.7mM KCl, 0.5mM MgCl2, 1.5mM CaCl2, 4mM HEPES, 11mM glucose), placed in a perfusion chamber, allowed to adhere to the coverslip, and then perfused with 37°C Tyrode buffer. The myocytes were stimulated at 1Hz and 10V. Only myocytes with clear texture, resting before pacing, cell length of 120-180 microns, baseline shortening fraction equal to 3-8% of the cell length, and contraction velocity greater than 100 microns / second were used for contractility experiments. The myocytes were first perfused with Tyrodes buffer for 60 seconds, followed by perfusion with the test compound for 5 minutes and washed with Tyrodes buffer for 140 seconds. Data were recorded continuously using IonOptix software and contractility data were analyzed using Ionwizard software (IonOptix). For each cell, 10-20 contractility transients were averaged and compared to baseline (no compound) conditions and conditions treated with compounds. Compound activity was measured by the effect on fractional shortening, where fractional shortening is the maximum length of the cell when contracted divided by the baseline cell length and normalized to 100% for untreated cells.

[0075] Table 2 Fractional shortening activation values ​​of the tested compounds

[0076]

[0077]

[0078] As shown in Table 2, relative to the baseline (no compound treatment) group, the fractional shortening activation value of compounds I-1 to I-3 at 1 μM was 65%, which was significantly better than the positive control MYK-491.

[0079] Example 6: Detection of pharmacokinetic properties of test compounds

[0080] Male SD rats were selected and the drug was administered orally (10 mg / kg) or intravenously (2 mg / kg). At 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, 10 h, and 24 h after administration, blood was continuously collected from the retinal venous plexus and placed in EP tubes containing heparin. The blood was centrifuged and the upper plasma was collected for LC-MS / MS analysis. Based on the blood drug concentration-time data obtained from the test, WinNonlin software was used to calculate the pharmacokinetic parameters and the oral bioavailability.

[0081] The results of the study showed that the oral bioavailability of MYK-491 in rats was 22% and the half-life was 2.0h; while the oral bioavailability of compound I-1 was increased to 65% and the half-life was 6.6h.

[0082] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is represented by any of the following structural formulas: 。 2. A pharmaceutical composition, characterized in that The pharmaceutical composition contains a therapeutically effective amount of the compound according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

3. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a cardiac myosin agonist.

4. Use of a compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating dilated cardiomyopathy, or a cardiac disease with pathophysiological characteristics of dilated cardiomyopathy, or a disease with contractile dysfunction or reduced contractile reserve.

5. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating diseases or conditions of systolic dysfunction, diastolic dysfunction, HFrEF, HFpEF, chronic heart failure and acute heart failure.

6. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating a disease or condition characterized by left ventricular systolic dysfunction or symptoms or reduced exercise capacity due to systolic dysfunction.

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