WO2019072130A1 - 1, 2, 4-triazole compound - Google Patents

Abstract

A 1,2,4-triazole compound shown in formula (I) and pharmaceutically acceptable salts thereof. The compound has the activity of an apoptosis signal-regulating kinase 1 ("ASK1") inhibitor; thus, the compound can be used for treating ASK1 mediated diseases including chronic liver disease, cardiovascular disease, metabolic disturbance, respiratory system disorder, gastrointestinal disorder, and neurodegenerative diseases. A pharmaceutical composition comprising the compound of formula (I) and uses thereof.

Description

1,2,4-triazole compound Technical field

The invention belongs to the technical field of medicine, and in particular relates to a 1,2,4-triazole compound and a composition comprising the same and use thereof. In particular, the invention relates to certain indole substituted 5-(4-cyclopropyl-1H-imidazol-1-yl)-2-fluoro-4-methyl-N-[6-(4-isopropyl- 4H-1,2,4,-triazol-3-yl)-2-pyridyl]-benzamide, these guanidine substituted compounds are used for the treatment of ASK1-mediated diseases with better pharmacokinetic properties .

Background technique

Many current drugs suffer from poor absorption, distribution, metabolism, and/or excretion (ADME) properties, which prevent them from being more widely used or limited to certain indications. Poor ADME properties are also an important cause of drug candidate failure in clinical trials. One such problem is rapid metabolism, which causes many drugs that would otherwise be highly effective in the treatment of disease to be cleared from the body too quickly. A common solution for rapid drug clearance is frequent or high dose administration to achieve sufficiently high plasma drug levels. However, this raises a number of potential therapeutic problems, such as poor patient compliance with medication regimens, increased side effects at higher doses, and increased treatment costs. Rapidly metabolized drugs can also expose patients to undesirable toxic or reactive metabolites.

Another ADME limitation that affects many drugs is the formation of toxic or bioreactive metabolites. Thus, some patients receiving the drug may be exposed to toxicity, or such drug safe doses may be limited such that the patient receives a suboptimal amount of active agent. In some cases, changing the dosing interval or formulation method may help reduce clinical adverse effects, but the formation of such undesirable metabolites is generally inherent in the metabolism of the compound.

One potentially attractive strategy for improving drug metabolism is 氘 modification. In this approach, attempts have been made to slow drug metabolism or reduce the formation of undesirable metabolites by replacing one or more hydrogen atoms with deuterium atoms. Radon is a safe, stable, non-radioactive hydrogen isotope. Tantalum forms a stronger bond with carbon than hydrogen. In the case of selection, an increase in bond strength imparted by hydrazone can positively affect the ADME properties of the drug, thereby creating the potential to increase drug efficacy, safety, and/or tolerance. At the same time, because the size and shape of the crucible are substantially the same as the size and shape of hydrogen, the replacement of hydrogen by deuterium is expected to not affect the biochemical potency and selectivity of the drug compared to the original chemical entity containing only hydrogen.

Over the past 35 years, the effect of hydrazine substitution on metabolic rate has been reported for a very small percentage of approved drugs (see, for example, Foster, AB, Adv Drug Res, 1985, 14: 1-40 ("Foster"); Fisher, MB Etc., Curr Opin Drug Discov Devel, 2006, 9: 101-09 ("Fisher)). The results are variable and unpredictable. For some compounds, deuteration causes a decrease in metabolic clearance in vivo. For other compounds, no metabolism Other compounds show an increase in metabolic clearance. The variability of the sputum effect also leads the skilled person to suspect or abandon the sputum modification as a viable drug design strategy for inhibitory metabolism (see Foster's 35 pages and Fisher's 101 page). ).

The effect of oxime modification on the metabolic properties of the drug is not predictable, even if the ruthenium atom penetrates into a known metabolic site. It is only possible to determine whether and how the metabolic rate differs from its non-deuterated counterpart by actually preparing and testing the deuterated drug. Many drugs have multiple sites where metabolism may occur. The degree of deuteration necessary for the various drugs will be different for sites that require deuteration and for the effects of metabolism (if any).

The present invention relates to novel derivatives of Selonsertib and pharmaceutically acceptable salts thereof. The invention also provides compositions comprising the compounds of the invention, and the use of such compositions in a method of treatment of diseases and conditions which are beneficially treated by administration of an ASK1 (apoptosis signal-regulated kinase 1) inhibitor.

Selonsertib, also known as GS-4997 and chemical name 5-(4-cyclopropyl-1H-imidazol-1-yl)-2-fluoro-4-methyl-N-[6-(4-isopropyl- 4H-1,2,4,-triazol-3-yl)-2-pyridine]-benzamide (having the structure shown below) is a small molecule inhibitor developed by Gilead Pharmaceuticals Co., Ltd., which can effectively reduce ASK1 Pathological function. The clinical phase II trial of Selonsertib in the treatment of nonalcoholic steatohepatitis (NASH) showed anti-fibrotic effects in NASH patients only after 24 weeks. At present, Selonsertib's research for the treatment of NASH is in clinical phase III, and the study for the treatment of alcoholic hepatitis is in clinical phase II.

Phosphorylation of the ASK1 protein can result in apoptosis or other cellular responses depending on the cell type. ASK1 activation and signaling have been reported to play an important role in a wide range of diseases including neurodegenerative disorders, cardiovascular disorders, inflammatory disorders and metabolic disorders. In addition, ASK1 is involved in mediating organ damage following ischemia and reperfusion of the heart, brain and kidney (Watanabe et al. (2005) BBRC 333, 562-567; Zhang et al. (2003) LifeSci 74-37-43; Terada Et al. (2007) BBRC364: 1043-49).

Even though Selonsertib is already present, there is a need for an effective compound with improved pharmacokinetic and/or pharmacodynamic performance in the treatment of ASKl activation related diseases.

Summary of the invention

In view of the above technical problems, the present invention discloses a 1,2,4-triazole compound and a composition comprising the same, which has a strong ASK1 inhibitor activity.

In this regard, the present invention adopts the following technical solutions:

A first aspect of the invention relates to a 1,2,4-triazole compound of the formula (I), or a pharmaceutically acceptable salt thereof:

among them,

R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Y ¹ , Y ² , Y ³ , Y ⁴ , Y ⁵ and Y ⁶ are each independently hydrogen or deuterium;

X ¹ , X ² and X ³ are each independently CH ₃ , CH ₂ D, CHD ₂ or CD ₃ .

Provided that if X ¹ , X ² and X ^{3 are} each CH ₃ , then R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Y ¹ , Y ² , Y ³ , at least one of Y ⁴ , Y ⁵ and Y ⁶ is 氘.

As a preferred embodiment of the present invention, the compound of the formula (I) contains at least one halogen atom, more preferably one germanium atom, more preferably two germanium atoms, more preferably three germanium atoms, more preferably four germanium atoms. More preferably, six helium atoms, more preferably seven helium atoms, more preferably nine helium atoms.

As a preferred embodiment of the invention, the cerium isotope content of cerium in the deuterated position is at least 0.015%, preferably greater than 30%, more preferably greater than 50%, more preferably greater than 75%, more preferably greater than the natural strontium isotope content. The ground is greater than 95%, more preferably greater than 99%.

Specifically, in the present invention, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Y ¹ , Y ² , Y ³ , Y ⁴ , Y ⁵ , Y ⁶ , X ¹ , X ² and X ³ having a strontium isotope content of at least 5%, preferably more than 10%, more preferably more than 15%, more preferably more than 20%, more preferably more than 25%, in each deuterated position, More preferably more than 30%, more preferably more than 35%, more preferably more than 40%, more preferably more than 45%, more preferably more than 50%, more preferably more than 55%, more preferably more than 60%, more Preferably more than 65%, more preferably more than 70%, more preferably more than 75%, more preferably more than 80%, more preferably more than 85%, more preferably more than 90%, more preferably more than 95%, more preferably The ground is greater than 99%.

In another specific embodiment, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Y ¹ , Y ² , Y ³ , Y ⁴ of the compound of formula (I) , Y ⁵ , Y ⁶ , X ¹ , X ² and X ³ , at least one of which contains ruthenium, more preferably two ruthenium, more preferably three ruthenium, more preferably four ruthenium, more preferably five More than six, more preferably six, more preferably seven, more preferably eight, more preferably nine, more preferably ten, more preferably eleven氘, preferably 12 氘, more preferably thirteen 氘, more preferably 14 氘, more preferably fifteen 氘, more preferably 16 氘, better The seventeenth place contains 氘, preferably the eighteen 氘, preferably nineteen 氘, more preferably twenty 氘, more preferably twenty one 氘, more preferably twenty two One contains 氘, and more preferably twenty-three. Specifically, the compound of the formula (I) contains at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, ten Three, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three atomic atoms.

As a preferred embodiment of the invention, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ and R ⁸ are each independently hydrogen or deuterium.

In another preferred embodiment, R ¹ , R ³ and R ⁴ are deuterium.

In another preferred embodiment, R ⁵ is deuterium.

In another preferred embodiment, R ¹ , R ³ , R ⁴ and R ⁵ are deuterium.

As a preferred embodiment of the invention, Y ¹ , Y ² , Y ³ , Y ⁴ , Y ⁵ and Y ⁶ are each independently hydrogen or deuterium.

In another preferred embodiment, Y ¹ is deuterium.

In another preferred embodiment, Y ² , Y ³ , Y ⁴ , Y ⁵ and Y ⁶ are deuterium.

As a preferred embodiment of the invention, X ¹ , X ² and X ³ are each independently CH ₃ , CH ₂ D, CHD ₂ or CD ₃ .

In another preferred embodiment, X ¹ , X ² is CD ₃ .

In another preferred embodiment, X ³ is CD ₃ .

In a preferred embodiment of the invention, the compound is selected from the group consisting of the compounds or pharmaceutically acceptable salts thereof:

In another preferred embodiment, the compound does not include a non-deuterated compound.

In a second aspect of the invention, the present invention also discloses a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a 1,2,4-triazole compound as described above, or a pharmaceutically acceptable compound thereof Accepted salt.

In a third aspect of the invention, the present invention also discloses a method for preparing a pharmaceutical composition as described above, comprising the steps of: pharmaceutically acceptable excipients with 1, 2, 4 -3 as described above The azole compound, or a pharmaceutically acceptable salt thereof, is mixed to form a pharmaceutical composition.

In another preferred embodiment, the pharmaceutical composition is an injection, a sachet, a tablet, a pill, a powder or a granule.

In another preferred embodiment, the pharmaceutical composition further comprises an additional therapeutic agent, the additional therapeutic agent being a medicament for treating cancer, cardiovascular disease, inflammation, infection, immune disease, metabolic disease or organ transplantation. .

A fourth aspect of the invention provides a method of treating at least a portion of a disease mediated by ASK1 in a patient in need thereof, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of the first aspect of the invention or a pharmaceutically acceptable compound thereof Accepted salt, or a pharmaceutical composition thereof.

In a particular embodiment, the compound treatable by the compounds of the invention is selected from the group consisting of diabetes, diabetic nephropathy, nephropathy, renal fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), liver fibrosis, pulmonary hypertension, non-alcoholic Fatty liver hepatitis, liver disease, alcoholic liver disease, inflammatory disease, autoimmune disease, proliferative disease, transplant rejection, diseases involving impaired cartilage metabolism, congenital cartilage malformation, or diseases associated with excessive secretion of IL6 .

In another preferred embodiment, the treatable disease is nonalcoholic steatohepatitis or alcoholic liver disease.

In another preferred embodiment, the treatable condition is pulmonary hypertension or pulmonary fibrosis.

In another preferred embodiment, the treatable disease is diabetic nephropathy, kidney disease or renal fibrosis.

It should be understood that within the scope of the present invention, the above various technical features, embodiments, and technical features specifically described in the following (such as the embodiments) may be combined with each other to constitute a new or preferred technical solution. . Due to space limitations, we will not repeat them here.

Detailed ways

The compound of the deuterated ASK1 inhibitor of the present invention and a pharmaceutically acceptable salt thereof have superior pharmacokinetic and/or pharmacodynamic properties compared to the undeuterated compound, and thus are more suitable as an ASK1 inhibitor The compounds, in turn, are more suitable for the preparation of a medicament for the treatment of ASK1-mediated related diseases. The present invention has been completed on this basis.

definition

As used herein, unless otherwise specified, "deuterated" means that one or more hydrogens in the compound or group are replaced by deuterium; deuteration may be monosubstituted, disubstituted, polysubstituted or fully substituted. The terms "one or more deuterated" are used interchangeably with "one or more deuterated".

As used herein, unless otherwise specified, "non-deuterated compound" means a compound containing a proportion of germanium atoms not higher than the natural helium isotope content (0.015%).

The invention also includes isotopically labeled compounds, equivalent to the original compounds disclosed herein. Examples of isotopes which may be listed as compounds of the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine isotopes such as ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁷ O, ¹⁸ O, respectively. , ³¹ P, ³² P, ³⁵ S, ¹⁸ F and ³⁶ Cl. a compound, or an enantiomer, a diastereomer, an isomer, or a pharmaceutically acceptable salt or solvate of the present invention, wherein an isotope or other isotopic atom containing the above compound is within the scope of the present invention . Certain isotopically-labeled compounds of the present invention, such as the radioisotopes of ³ H and ¹⁴ C, are also among them, useful in tissue distribution experiments of drugs and substrates.氚, ie ³ H and carbon-14, ie ¹⁴ C, are easier to prepare and detect and are preferred in isotopes. Isotopically labeled compounds can be prepared in a conventional manner by substituting a readily available isotopically labeled reagent with a non-isotopic reagent using the protocol of the examples.

Compound

The present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, crystal form, stereoisomer or isotopic variation thereof:

among them,

R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Y ¹ , Y ² , Y ³ , Y ⁴ , Y ⁵ , Y ⁶ are each independently hydrogen or hydrazine;

X ¹ , X ² , X ³ are each independently CH ₃ , CH ₂ D, CHD ₂ or CD ₃ ;

Provided that if X ¹ , X ² and X ^{3 are} each CH ₃ , then R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Y ¹ , Y ² , Y ³ , at least one of Y ⁴ , Y ⁵ and Y ⁶ is 氘.

As a preferred embodiment of the invention, the cerium isotope content of cerium in the deuterated position is at least 0.015% greater than the natural isotope content, preferably greater than 30%, more preferably greater than 50%, more preferably greater than 75%, more preferably Preferably, it is greater than 95%, more preferably greater than 99%.

In a particular embodiment of formula (I), "R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ and R ⁸ are each independently hydrogen or deuterium" includes R ¹ selected from Hydrogen or hydrazine, R ^{2 is} selected from hydrogen or hydrazine, R ^{3 is} selected from hydrogen or hydrazine, and so on, until R ^{8 is} selected from hydrogen or hydrazine. More specifically, R ¹ is hydrogen or R ¹ is deuterium, R ² is hydrogen or R ² is deuterium, R ³ is hydrogen or R ³ is deuterium, and so on, until R ⁸ is hydrogen or R ⁸ is deuterium. Technical solutions.

In another specific embodiment of formula (I), "Y ¹ , Y ² , Y ³ , Y ⁴ , Y ⁵ and Y ⁶ are each independently hydrogen or deuterium" includes Y ^{1 being} selected from hydrogen or deuterium, Y ^{2 is} selected from hydrogen or hydrazine, Y ^{3 is} selected from hydrogen or hydrazine, and so on, until Y ^{6 is} selected from hydrogen or hydrazine. More specifically, it includes that Y ¹ is hydrogen or Y ¹ is deuterium, Y ² is hydrogen or Y ² is deuterium, Y ³ is hydrogen or Y ³ is deuterium, and so on until Y ⁶ is hydrogen or Y ⁶ is deuterium. Technical solutions.

In another specific embodiment of formula (I), "X ¹ , X ² , X ³ are each independently CH ₃ , CH ₂ D, CHD ₂ or CD ₃ " includes X ¹ selected from CH ₃ , CH ₂ D, CHD ₂ or CD ₃ , X ^{2 is} selected from the group consisting of CH ₃ , CH ₂ D, CHD ₂ or CD ₃ , and X ^{3 is} selected from the group consisting of CH ₃ , CH ₂ D, CHD ₂ or CD ₃ . More specifically, it includes that X ¹ is CH ₃ , X ¹ is CH ₂ D, X ¹ is CHD ₂ or X ¹ is CD ₃ , X ² is CH ₃ , X ² is CH ₂ D, and X ² is CHD ₂ or X. ² is a technical scheme in which CD ₃ , X ³ is CH ₃ , X ³ is CH ₂ D, X ³ is CHD ₂ or X ³ is CD ₃ .

In a preferred embodiment, the invention relates to a compound of formula (I), or a pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, crystalline form, stereoisomer or isotopic variation thereof, wherein R ⁷ And R ⁸ , Y ² -Y ⁶ are hydrogen, R ¹ -R ⁸ and Y ¹ are each independently hydrogen or deuterium, and X ¹ , X ² and X ³ are each independently CH ₃ , CH ₂ D, CHD ₂ or CD _{3 is} additionally provided that the compound contains at least one ruthenium atom.

In another preferred embodiment, R ² and R ⁶ are hydrogen.

In another preferred embodiment, R ¹ , R ³ and R ⁴ are each independently selected from hydrogen or hydrazine. In another more preferred embodiment, R ¹ , R ³ and R ⁴ are hydrogen. In another more preferred embodiment, R ¹ , R ³ and R ⁴ are deuterium.

In another preferred embodiment, R ⁵ is hydrogen or deuterium. In another more preferred embodiment, R ⁵ is hydrogen. In another more preferred embodiment, R ⁵ is deuterium.

In another preferred embodiment, X ³ is CH ₃ .

In another preferred embodiment, X ¹ and X ² are each independently selected from CH ₃ or CD ₃ . In another more preferred embodiment, X ¹ and X ² are CH ₃ . In another more preferred embodiment, X ¹ and X ² are CD ₃ .

In another preferred embodiment, Y ¹ is hydrogen or deuterium. In another more preferred embodiment, Y ¹ is hydrogen. In another more preferred embodiment, Y ¹ is hydrazine.

Wherein the term "pharmaceutically acceptable salt" means that it is suitable for contact with human and lower animal tissues without undue toxicity, irritation, allergies, etc., and with reasonable benefits, within the scope of sound medical judgment. / Those dangerous proportions of those salts. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al., pharmaceutically acceptable salts as described in detail in J. Pharmaceutical Sciences (1977) 66: 1-19. Pharmaceutically acceptable salts of the compounds of the invention include those derived from suitable inorganic and organic acids and inorganic and organic bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or salts with organic acids such as acetic acid, oxalic acid, Maleic acid, tartaric acid, citric acid, succinic acid or malonic acid. Also included are salts formed using conventional methods in the art, for example, ion exchange methods. Other pharmaceutically acceptable salts include: adipic acid salts, alginate, ascorbate, aspartate, besylate, benzoate, disulfate, borate, butyrate, camphor Acid salt, camphor sulfonate, citrate, cyclopentanoate, digluconate, lauryl sulfate, ethanesulfonate, formate, fumarate, gluconate, glycerol Phosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate , malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate Salt, pectin ester, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, Thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Pharmaceutically acceptable salts derived from suitable bases include the alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium salts, and the like. If appropriate, other pharmaceutically acceptable salts include non-toxic ammonium salts, quaternary ammonium salts and amine cations formed with counterions, counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, Nitrate, lower alkyl sulfonate and aryl sulfonate.

The term "solvate" refers to a complex of a compound of the invention that is coordinated to a solvent molecule to form a specific ratio. "Hydrate" means a complex formed by the coordination of a compound of the invention with water.

The term "prodrug" includes a compound of the formula (I) which is biologically active or inactive, which, when taken by a suitable method, is metabolized or chemically reacted in the human body, or converted into a compound of the formula (I), or a formula a salt or solution of a compound of (I). The prodrugs include, but are not limited to, carboxylic acid esters, carbonates, phosphates, nitrates, sulfates, sulfone esters, sulfoxide esters, amino compounds, carbamates, azo compounds, phosphorus of the compound. In the form of an amide, a glucoside, an ether, an acetal or the like.

Pharmaceutical composition and method of administration

Since the compound of the present invention has excellent activity for inhibiting ASK1 kinase, the compound of the present invention and various crystal forms thereof, pharmaceutically acceptable salts, hydrates or solvates thereof, and a pharmaceutical composition containing the compound of the present invention as a main active ingredient Therapies can be used to treat, prevent, and alleviate ASK1-mediated diseases. According to the prior art, the compounds of the present invention are useful for the treatment of chronic liver diseases, cardiovascular diseases, metabolic disorders, respiratory disorders, gastrointestinal disorders, and neurodegenerative diseases.

The pharmaceutical compositions of the present invention comprise a safe or effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier. By "safe and effective amount" it is meant that the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. In general, the pharmaceutical compositions contain from 0.5 to 2000 mg of the compound of the invention per agent, more preferably from 1 to 500 mg of the compound of the invention per agent. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable excipient" means a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound formulated together. Pharmaceutically acceptable carriers, adjuvants, or vehicles that can be used in the compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (eg, human serum albumin) ), buffer substances (such as phosphate), glycine, sorbic acid, potassium sorbate, a mixture of partial glycerides of saturated plant fatty acids, water, salt or electrolyte (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, Sodium chloride, zinc salt, silica gel, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based material, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, wax, polyethylene-polyoxypropylene-embedded Segment polymer, polyethylene glycol and lanolin.

The mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, oral, duodenal, rectal, parenteral (intravenous, intramuscular or subcutaneous) and topical administration. medicine.

Solid dosage forms for oral administration include capsules, tablets, pills, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or solubilizer, for example, starch , lactose, sucrose, glucose, dry diol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectants, For example, glycerin; (d) a disintegrant such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent, for example, paraffin; An absorption accelerator, for example, a quaternary amine compound; (g) a wetting agent such as cetyl alcohol and glyceryl monostearate; (h) an adsorbent, for example, kaolin; and (i) a lubricant, for example, talc , calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or a mixture thereof. In capsules, tablets and pills, the dosage form may also contain a buffer.

Solid dosage forms such as tablets, troches, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other materials known in the art. They may contain opacifying agents and the release of the active compound or compound in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials. If necessary, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs. Liquid dosage forms can contain, in addition to the active compound, inert release agents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances.

In addition to these inert diluents, the compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.

In addition to the active compound, the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.

Compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.

Dosage forms for the compounds of the invention for topical administration include ointments, powders, patches, propellants and inhalants. The active ingredient is admixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or, if necessary, propellants.

The compounds of the invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

When a pharmaceutical composition is used, a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective effective dosage, for a 60 kg body weight, The dose to be administered is usually 0.5 to 2000 mg, preferably 1 to 500 mg. Of course, specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.

Method of treating disease

Therapeutic agents useful as inhibitors of ASK1 signaling have the potential to treat or improve the life of neurodegenerative diseases, cardiovascular diseases, inflammation, autoimmune diseases, and metabolic disorders in patients in need of treatment for a disease or condition. In particular, ASK1 inhibitors have the potential to treat heart and kidney disease, including kidney disease, diabetic nephropathy, chronic kidney disease, fibrotic diseases (including lung and kidney fibrosis), dilated cardiomyopathy, respiratory diseases (including chronic obstruction). Severe lung disease (COPD) and acute lung injury), acute and chronic liver disease (such as nonalcoholic steatohepatitis and alcoholic hepatitis).

A variety of assays for identifying the ability of a compound to inhibit ASK1 kinase activity and its effectiveness as an ASK1 inhibitor are known in the art and are described, for example, in U.S. Patent No. 8,742,126.

Some embodiments described herein relate to the use of a compound described herein or a pharmaceutical composition described herein for treating a disease in a patient in need of treatment with an ASK1 inhibitor.

Some embodiments described herein are methods of treating diabetic nephropathy or diabetic complications comprising administering a therapeutically effective amount of a compound of formula (I) described herein or a pharmaceutical composition described herein. In some embodiments, diabetes includes type 1 and type 2 diabetes, gestational diabetes, pre-diabetes, insulin resistance, metabolic syndrome, impaired fasting glucose, and impaired glucose tolerance. Type 1 diabetes is also known as insulin-dependent diabetes mellitus (IDDM). Type 2 is also known as non-insulin-dependent diabetes mellitus (NIDDM).

Another embodiment is directed to a method of treating a kidney disease or diabetic nephropathy comprising administering a therapeutically effective amount of a compound of formula (I) as described herein or a pharmaceutical composition described herein.

Another embodiment is directed to a method of treating renal fibrosis, pulmonary fibrosis, or idiopathic pulmonary fibrosis (IPF) comprising administering a therapeutically effective amount of a form of Compound I described herein or a pharmaceutical composition described herein.

Another embodiment is directed to a method of treating diabetic nephropathy, diabetic nephropathy, renal fibrosis, liver fibrosis or pulmonary fibrosis comprising administering a therapeutically effective amount of a crystalline form of Compound I described herein or a pharmaceutical combination as described herein Things.

Disclosed herein are methods of treating and/or preventing liver disease in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a form of Compound I described herein, or a composition thereof, optionally in combination with a therapeutically effective amount of a LOXL2 inhibitor. . The presence of active liver disease can be detected by the presence of elevated levels of enzymes in the blood. In particular, blood levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), which are known to be higher than the clinically accepted normal range, indicate ongoing liver damage. Routine monitoring of blood levels of ALT and AST in patients with liver disease is clinically used to measure the progression of liver disease at the time of medical treatment. Decreasing elevated ALT and AST to the normal range of acceptance as clinical evidence reflects a reduction in the severity of ongoing liver damage in patients.

In certain embodiments, the liver disease is chronic liver disease. Chronic liver disease involves progressive destruction and regeneration of the liver parenchyma, leading to fibrosis and cirrhosis. In general, chronic liver disease can be caused by viruses (such as hepatitis B, hepatitis C, cytomegalovirus (CMV) or Epstein Barr virus (EBV)), toxic agents or drugs (such as alcohol, methotrexate or nitrofurantoin), metabolism. Sexual diseases (eg nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), hemochromatosis or Wilson's disease), autoimmune diseases (eg autoimmune chronic hepatitis, primary biliary Cirrhosis or primary sclerosing cholangitis) or other causes (such as right heart failure).

In a specific embodiment, the liver disease is metabolic liver disease. In one embodiment, the liver disease is nonalcoholic fatty liver disease (NAFLD). NAFLD is associated with insulin resistance and metabolic syndrome (obesity, mixed hyperlipidemia, diabetes (type II), and hypertension). It is believed that NAFLD covers a range of disease activities and begins to accumulate fat (hepatic steatosis) in the liver.

The compounds of the present invention have a number of advantages over non-deuterated compounds known in the art. Advantages of the present invention include: First, the compound using the technical scheme of the present invention has excellent inhibitory effect on ASK1 protein kinase. Second, the metabolism of the compound in the organism is improved, giving the compound better pharmacokinetic parameter characteristics. In this case, the dosage can be changed and a long-acting preparation can be formed to improve the applicability. Third, the drug concentration of the compound in the animal is increased, and the drug efficacy is improved. Fourth, certain metabolites are inhibited and the safety of the compounds is increased.

Example

The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually in accordance with conventional conditions or according to the conditions recommended by the manufacturer. Parts and percentages are parts by weight and percentage by weight unless otherwise stated.

Usually, in the preparation scheme, each reaction is usually carried out in an inert solvent at room temperature to reflux temperature (e.g., 0 ° C to 100 ° C, preferably 0 ° C to 80 ° C). The reaction time is usually from 0.1 to 60 hours, preferably from 0.5 to 24 hours.

Example 1 5-(4-Cyclopropyl-1H-imidazolyl-1-yl)-N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl-5) -d)pyridin-2-yl Preparation of -3,4-d ₂ )-2-fluoro-4-methylbenzamide (Compound T-1).

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 2.

Compound 1 (5.0 g, 32.86 mmol) and methanol (60 mL) were added to a 100 mL one-necked flask equipped with magnetic stirring, and the mixture was stirred and dissolved, and hydrazine hydrate (3.29 g, 65.72 mmol) was slowly added dropwise. After the dropwise addition, the reaction was mixed. The liquid was heated under reflux for 3 hours, then cooled to room temperature, and a large amount of white solid was precipitated, filtered, and filtered, washed with cold methanol and dried to yield 3.5 g of white solid. LC-MS (APCI): m / z = 153.2 (M + 1) + 1 H NMR (DMSO-d 6, 300MHz) (δ / ppm):. 9.14 (s, 1H), 7.51 (t, J = 5.7 Hz, 1H), 7.11 (d, J = 5.7 Hz, 1H), 6.61 (d, J = 6.0 Hz, 1H), 6.08 (s, 2H), 4.48 (s, 2H).

Step 2 Synthesis of Compound 3.

Compound 2 (3.5 g, 23 mmol), toluene (Tol, 50 mL), isopropylamine (9.8 g, 166 mmol) and N,N-dimethylformamide-dipropyl were added to a 100 mL one-neck flask with magnetic stirring. Acetal (DMF-DPA, 10.89 g, 62 mmol), acetic acid (2.1 g, 35 mmol) was slowly added dropwise with stirring, and the reaction mixture was heated under reflux for 24 h under N ₂ atmosphere. After cooling to room temperature, the solvent was evaporated under reduced pressure, water (40 mL) was evaporated, and a large solid was precipitated with stirring, and filtered, and the filter cake was washed with isopropyl alcohol to obtain a white solid (3.1 g, yield: 66.3%). LC-MS (APCI): m / z = 204.2 (M + 1) + 1 H NMR (CDCl 3, 300MHz) (δ / ppm):. 8.31 (s, 1H), 7.58-7.54 (m, 2H), 6.56 (dd, J ₁ = 5.4 Hz, J ₂ = 1.5 Hz, 1H), 5.64 - 5.57 (m, 1H), 4.45 (s, 2H), 1.52 (d, J = 4.8 Hz, 6H).

Step 3 Synthesis of Compound 4.

Compound 3 (200 mg, 984 umol), heavy water (10 mL) and Pd/C (50 mg, 10%) were added to a 50 mL sealed tube equipped with magnetic stirring. The mixture was bubbled with hydrogen for 5 minutes while stirring, and then heated to 110 ° C after sealing. And stir the reaction overnight. After cooling to room temperature, dichloromethane (20 mL) was added, filtered, EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj The yield was 59.1%. LC-MS (APCI): m / z = 207.3 (M + 1) + 1 H NMR (CDCl 3, 300MHz) (δ / ppm):. 7.56 (s, 1H), 5.62-5.59 (m, 1H), 4.50 (s, 1H), 1.51 (d, J = 5.4 Hz, 6H).

Step 4 Synthesis of Compound 7.

Compound 6 (5.0 g, 24.5 mmol) and toluene (50 mL) were added to a 100 mL single-necked flask equipped with magnetic stirring, and the mixture was stirred and dissolved. Compound 5 (4.4 g, 27.0 mmol) and N,N-diisopropyl were added dropwise. Ethylethylamine (DIPEA, 8.5 mL, 51.5 mmol). Cooled to room temperature, water (50 mL), layers were separated and the organic phase was washed with saturated aqueous NH ₄ Cl (20mL), saturated aqueous NaHCO ₃ (20mL) and brine (20mL), dried over anhydrous sodium sulfate, Filtration and concentration, the residue was recrystallized from n-hexane, filtered, and dried to give a white solid. LC-MS (APCI): m / z = 286.2 & 288.2 (M + 1) + 1 H NMR (DMSO-d 6, 300MHz) (δ / ppm):. 7.07 (d, J = 6.9Hz, 1H) , 6.52 (d, J = 4.5 Hz, 1H), 5.29 (t, J = 4.2 Hz, 1H), 4.18 (d, J = 4.2 Hz, 2H), 4.52-4.51 (m, 1H), 2.11 (s, 3H), 0.98-0.88 (m, 4H).

Step 5 Synthesis of Compound 8.

Compound 7 (3.1 g, 10.8 mmol) and dichloromethane (DCM, 15 mL) were added to a 50 mL single-necked flask with magnetic stirring, and the mixture was stirred and cooled to 0 ° C, and formic acid (15 mL) and acetic anhydride were slowly added dropwise. (Ac ₂ O, 4.1 mL, 43.3 mmol), and the reaction mixture was stirred at 0 ° C for 3 hours under N ₂ atmosphere. Add water (20mL) to quench the reaction, adjust the pH to 9 with 40% NaOH water, separate the organic phase, extract the aqueous phase with dichloromethane (30mLx2), combine the organic phase, dry over anhydrous sodium sulfate, filter, concentrate directly In the next step. LC-MS (APCI): m / z = 314.1 & 316.1 (M + 1) + 1 H NMR (DMSO-d 6, 300MHz) (δ / ppm):. 8.15 (d, J = 9.6Hz, 1H) , 7.61 (d, J = 5.4 Hz, 1H), 7.41 (d, J = 7.2 Hz, 1H), 4.68 (d, 2H), 2.50 (s, 3H), 2.12-1.98 (m, 1H), 0.96- 0.85 (m, 4H).

Step 6 Synthesis of Compound 9.

To turn 50mL one-necked flask equipped with a magnetic stirrer was added Compound 8 (3.39g, 10.8mmol) and glacial acetic acid (30 mL), stirred clear solution was added ammonium acetate (2.57g, 42.1mmol), N ₂ atmosphere reaction mixture Heat to reflux overnight. After cooling to room temperature, the acetic acid was evaporated under reduced pressure, water (20 mL), 40% NaOH aqueous solution was adjusted to pH 9 and dichloromethane (30mL×3), organic phase was combined, dried over anhydrous sodium sulfate, filtered, concentrated, residue The white solid was separated by a silica gel column, 1.7 g, yield 53.4%. LC-MS (APCI): m / z = 295.1 and ^{297.1 (M + 1) + 1} H NMR (CDCl 3, 300MHz) (δ / ppm):. 7.41 (d, J = 4.8Hz, 1H), 7.37 ( s, 1H), 7.07 (d, J = 6.6 Hz, 1H), 6.72 (s, 1H), 2.14 (s, 3H), 1.90 - 1.85 (m, 1H), 0.90 - 0.78 (m, 4H).

Step 7 Synthesis of Compound 10.

50mL three-necked flask equipped with a magnetic stirrer was added Compound 9 (1.7g, 5.8mmol), evacuated and replaced three times with N _2, N ₂ atmosphere was added dropwise under anhydrous THF (30mL), a clear solution was stirred, cooled to 0 °C, slowly add isopropylmagnesium chloride (4.3mL, 8.6mmol, 2M), stir and react for 1 hour, then slowly introduce CO ₂ into the reaction solution through a balloon filled with CO ₂ gas, react for 1 hour, add water ( 20mL) quenched the reaction, 6M HCl aqueous solution was adjusted to pH 5, ethyl acetate extracted (30mL×3), and the organic phase was combined, dried over anhydrous sodium sulfate, filtered, concentrated, crystals %. LC-MS (APCI): m / z = 261.1 (M + 1) + 1 H NMR (DMSO-d 6, 300MHz) (δ / ppm):. 9.17 (s, 1H), 7.95 (d, J = 4.8 Hz, 1H), 7.70 (s, 1H), 7.52 (d, J = 8.4 Hz, 1H), 2.24 (s, 3H), 2.05-1.91 (m, 1H), 1.01-0.85 (m, 4H).

Step 8 Synthesis of Compound T-1.

50mL three-necked flask equipped with a magnetic stirrer was added Compound 10 (100mg, 0.384mmol), evacuated and replaced with N _2, under N ₂ atmosphere through a syringe dropwise dry dichloromethane (2mL) and of DMF (2mL), stirred The solution was dissolved, and the reaction solution was cooled to 0 ° C, and oxalyl chloride (0.33 mL, 0.653 mmol, 2M in dichloromethane) was slowly added dropwise, and the mixture was dropped, the ice bath was removed, and the reaction was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure and dichloromethane was added dry dichloromethane (10 mL), under N ₂ atmosphere clear solution was stirred and cooled to 0 deg.] C, was slowly added dropwise Compound 4 (94mg, 0.461mmol) in (2 mL), followed by the dropwise DIPEA (0.19 mL, 1.15 mmol) was added, the ice bath was removed, and the mixture was stirred at room temperature for 2 hr. The reaction was quenched with water (20 mL), EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj The yield was 35.1%. LC-MS (APCI): m / z = 446.4 (M + 1) + 1 H NMR (CDCl 3, 300MHz) (δ / ppm):. 9.06 (d, J = 6.0Hz, 1H), 8.40-8.36 ( m, 2H), 8.07-8.05 (m, 2H), 7.91 (t, J = 6.0 Hz, 1H), 7.45 (s, 1H), 7.18 (d, J = 9.0 Hz, 1H), 6.78 (s, 1H) ), 5.51-5.44 (m, 1H), 2.28 (s, 3H), 1.91-1.88 (m, 1H), 1.59 (d, J = 5.1 Hz, 6H), 0.90-0.81 (m, 4H).

Example 2 5-(4-Cyclopropyl-1H-imidazolyl-1-yl)-N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridine -2-yl)-2-fluoro-4- Preparation of methylbenzamide-6-d (Compound T-2).

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 12.

Add compound 6 (2.0 g, 9.8 mmol) and heavy water (10 mL) to a 20 mL microwave tube equipped with magnetic stirring, and slowly add a heavy aqueous solution of DCl (0.817 mL, 9.8 mmol, 12 M) under stirring, add the reaction mixture, microwave The temperature was raised to 160 ° C and the reaction was carried out for 1.5 hours. Cooled to room temperature, saturated NaHCO ₃ adjusted to pH 10, extracted with dichloromethane (20 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give 1.59 g of a brown solid, yield 79.1%. LC-MS (APCI): m / z = 205.1 (M + 1) + 1 H NMR (DMSO-d 6, 300MHz) (δ / ppm):. 6.93 (d, J = 7.2Hz, 1H), 4.94 ( s, 2H), 1.99 (s, 3H).

Step 2 Synthesis of Compound 13.

Compound 12 (1.59 g, 7.75 mmol) and toluene (30 mL) were added to a 100 mL single-necked flask with magnetic stirring, and the mixture was stirred and dissolved. Compound 5 (1.39 g, 8.53 mmol) and DIPEA (2.7 mL, 16.28 mmol) were added dropwise. The reaction solution was heated to reflux for 2 hours. Cooled to room temperature, water (30 mL), layers were separated and the organic phase was washed with saturated aqueous NH ₄ Cl (10 mL), saturated aqueous NaHCO ₃ (10mL) and saturated saline water (10mL), dried over anhydrous sodium sulfate, Filtration and concentration, the residue was recrystallized from n-hexane, filtered and dried to give a white solid, 1.5 _g , yield 67.4%. LC-MS (APCI): m/z = 287.2 & 289.2 (M + 1) ⁺ .

Step 3 Synthesis of Compound 14.

Compound 13 (1.5 g, 5.22 mmol) and dichloromethane (10 mL) were added to a 50 mL single-necked flask with magnetic stirring, and the mixture was stirred and cooled to 0 ° C, and formic acid (15 mL) and acetic anhydride were slowly added dropwise ( 2.1 mL, 20.9 mmol), and the reaction solution was stirred at 0 ° C for 3 hours under N ₂ atmosphere. Add water (10mL) to quench the reaction, adjust the pH to 9 with 40% NaOH water, separate the organic phase, extract the organic phase with dichloromethane (20mLx2), combine the organic phase, dry over anhydrous sodium sulfate, filter, concentrate directly In the next step. LC-MS (APCI): m/z = 315.1 & 317.1 (M + 1) ⁺ .

Step 4 Synthesis of Compound 15.

Compound 14 (1.65 g, 5.24 mmol) and glacial acetic acid (15 mL) were added to a 50 mL single-necked flask equipped with magnetic stirring, and the mixture was stirred and dissolved. Ammonium acetate (1.25 g, 20.5 mmol) was added, and the reaction mixture was heated under N ₂ atmosphere. Reflux overnight. After cooling to room temperature, the acetic acid was evaporated under reduced pressure, water (10 mL), 40% NaOH aqueous solution was adjusted to pH 9 and extracted with dichloromethane (20mL×3). The organic phase was combined, dried over anhydrous sodium sulfate and filtered. The column was separated into 1.7 g of a white solid, yield 53.4%. LC-MS (APCI): m / z = 296.1 and ^{298.1 (M + 1) + 1} H NMR (CDCl 3, 300MHz) (δ / ppm):. 7.38 (s, 1H), 7.09 (d, J = 6.9 Hz, 1H), 6.73 (s, 1H), 2.15 (s, 3H), 1.91-1.87 (m, 1H), 0.91-0.78 (m, 4H).

Step 5 Synthesis of Compound 16.

50mL three-necked flask equipped with a magnetic stirrer was added Compound 15 (820mg, 2.77mmol), evacuated and replaced three times with N _2, N ₂ atmosphere was added dropwise under anhydrous THF (15mL), stirred clear solution was cooled to 0 ℃ , was slowly added dropwise isopropylmagnesium chloride (2.77mL, 5.54mmol, 2M), incubated for 1 hour with stirring, and then slowly into the reaction mixture into CO ₂ by the CO ₂ gas balloon is filled, for 1 hour, water (20 mL The reaction was quenched, and the mixture was evaporated to mjjjjjjjjjjjjjs %. LC-MS (APCI): m / z = 262.1 (M + 1) + 1 H NMR (DMSO-d 6, 300MHz) (δ / ppm):. 8.69 (s, 1H), 7.53 (s, 1H), 7.50 (d, J = 8.7 Hz, 1H), 2.24 (s, 3H), 1.96-1.93 (m, 1H), 0.96-0.93 (m, 2H), 0.80-0.77 (m, 2H).

Step 6 Synthesis of Compound T-2.

50mL three-necked flask equipped with a magnetic stirrer was added Compound 16 (130mg, 0.497mmol), evacuated and replaced three times with N _2, were added dropwise under N ₂ atmosphere through a syringe of dry dichloromethane (2mL) and DMF (2mL) The mixture was stirred and stirred, and the reaction solution was cooled to 0 ° C. oxalyl chloride (0.42 mL, 0.846 mmol, 2M in dichloromethane) was slowly added dropwise, and the mixture was evaporated, and the reaction was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, added dry dichloromethane (10 mL), under N ₂ atmosphere clear solution was stirred and cooled to 0 deg.] C, was slowly added dropwise compound 3 (122mg, 0.597mmol) in dichloromethane (2 mL), followed by the dropwise DIPEA (0.25 mL, 1.49 mmol) was added, the ice bath was removed, and the mixture was stirred at room temperature for 2 hours. The reaction was quenched with water (20 mL), EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj The yield was 35.1%. LC-MS (APCI): m / z = 447.4 (M + 1) + 1 H NMR (CDCl 3, 300MHz) (δ / ppm):. 9.06 (d, J = 12.3Hz, 1H), 8.41-8.38 ( m, 2H), 8.07 (d, J = 5.7 Hz, 1H), 7.93 (t, J = 6.0 Hz, 1H), 7.49 (s, 1H), 7.20 (d, J = 9.6 Hz, 1H), 6.78 ( s, 1H), 5.52-5.46 (m, 1H), 2.29 (s, 3H), 2.00-1.88 (m, 1H), 1.59 (d, J = 5.1 Hz, 6H), 0.91 - 0.81 (m, 4H) .

Example 3 5-(4-Cyclopropyl-1H-imidazolyl-1-yl)-N-(6-(4-(propan-2-yl-1,1,1,3,3,3-d) ₆ )-4H-1,2,4-triazol-3-yl)pyridin Preparation of pyridin-2-yl)-2-fluoro-4-methylbenzamide (Compound T-3).

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 18.

Ammonium acetate (6.32 g, 78 mmol) and MeOD (50 mL) were added to a 50 mL one-neck flask equipped with magnetic stirring, and the mixture was refluxed for 3 hours under N ₂ atmosphere. Concentrated under reduced pressure to dryness, then added MeOD (10mL), was added with stirring acetone -d ₆ (1.0g, 15.6mmol) and _{NaBH 3 CN (980mg, 15.6mmol)} , stirred at room temperature under N ₂ atmosphere overnight. Add water (10mL) to quench the reaction, adjust the pH to 2 with 6M hydrochloric acid, extract with ethyl acetate (20mLx2), adjust the pH to 12 with water 6M NaOH, extract with dichloromethane (20mLx3), combine the organic phase, anhydrous sulfuric acid The sodium was dried, filtered, and the filtrate was adjusted to pH 2 with 2M hydrochloric acid methanol and concentrated to dryness to give compound 18 hydrochloride.

Step 2 Synthesis of Compound 19.

Compound 2 (300 mg, 1.97 mmol), toluene (10 mL), Compound 18 hydrochloride (642 mg, 9.86 mmol) and N,N-dimethylformamide-dipropyl were added to a 50 mL one-neck flask with magnetic stirring. The acetal (933 mg, 5.32 mmol) was slowly added dropwise with stirring (296 mg, 4.93 mmol), and the mixture was heated and refluxed for 3 hours under N ₂ atmosphere. The mixture was cooled to room temperature, and the solvent was evaporated under reduced pressure. LC-MS (APCI): m / z = 210.2 (M + 1) + 1 H NMR (CDCl 3, 300MHz) (δ / ppm):. 8.32 (s, 1H), 7.62-7.56 (m, 2H), 6.58 (dd, J ₁ =5.1 Hz, J ₂ =1.2 Hz, 1H), 5.59 (s, 1H), 4.51 (s, 2H).

Step 3 Synthesis of Compound T-3.

50mL three-necked flask equipped with a magnetic stirrer was added Compound 10 (130mg, 0.497mmol), evacuated and replaced with N _2, were added dropwise under N ₂ atmosphere through a syringe of dry dichloromethane (2mL) and of DMF (2mL), stirring a clear solution, the reaction solution was cooled to 0 deg.] C, was slowly added dropwise oxalyl chloride (0.42mL, 0.846mm _o l, 2M in dichloromethane) dropwise, remove the ice bath, stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, added dry dichloromethane (10 mL), under N ₂ atmosphere clear solution was stirred and cooled to 0 deg.] C, was slowly added dropwise Compound 19 (122mg, 0.597mmol) in dichloromethane (2 mL), followed by the dropwise DIPEA (0.25 mL, 1.49 mmol) was added, the ice bath was removed, and the mixture was stirred at room temperature for 2 hours. The reaction was quenched with water (20 mL), EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj The yield was 35.1%. LC-MS (APCI): m / z = 452.4 (M + 1) + 1 H NMR (CDCl 3, 300MHz) (δ / ppm):. 9.05 (d, J = 11.7Hz, 1H), 8.39-8.35 ( m, 2H), 8.07-8.04 (m, 2H), 7.91 (t, J = 6.6 Hz, 1H), 7.45 (s, 1H), 7.18 (d, J = 9.0 Hz, 1H), 6.77 (s, 1H) ), 5.43 (s, 1H), 2.27 (s, 3H), 1.94-1.90 (m, 1H), 0.90-0.80 (m, 4H).

Example 4 5-(4-Cyclopropyl-1H-imidazolyl-1-yl)-N-(6-(4-(propan-2-yl-d ₇ )-4H-1,2,4-tri Zyrid-3-yl)pyridin-2-yl)-2- Preparation of fluoro-4-methylbenzamide (Compound T-4).

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 20.

Ammonium acetate (6.32 g, 78 mmol) and MeOD (50 mL) were added to a 50 mL one-neck flask equipped with magnetic stirring, and the mixture was refluxed for 3 hours under N ₂ atmosphere. Concentrated under reduced pressure to dryness, then added MeOD (10mL), was added with stirring acetone -d ₆ (1.0g, 15.6mmol) and _{NaBD 3 CN (980mg, 15.6mmol)} , stirred at room temperature under N ₂ atmosphere overnight. Add water (10mL) to quench the reaction, adjust the pH to 2 with 6M hydrochloric acid, extract with ethyl acetate (20mLx2), adjust the pH to 12 with water 6M NaOH, extract with dichloromethane (20mLx3), combine the organic phase, anhydrous sulfuric acid The sodium was dried, filtered, and the filtrate was adjusted to pH 2 with 2M hydrochloric acid methanol and concentrated to dryness to give Compound 20 hydrochloride.

Step 2 Synthesis of Compound 21.

Compound 2 (300 mg, 1.97 mmol), toluene (10 mL), Compound 20 hydrochloride (642 mg, 9.86 mmol) and N,N-dimethylformamide-dipropyl were added to a 50 mL one-neck flask with magnetic stirring. The acetal (933 mg, 5.32 mmol) was slowly added dropwise with stirring (296 mg, 4.93 mmol), and the mixture was heated and refluxed for 3 hours under N ₂ atmosphere. The mixture was cooled to room temperature, and the solvent was evaporated under reduced pressure. LC-MS (APCI): m / z = 211.2 (M + 1) +.

Step 3 Synthesis of Compound T-4.

50mL three-necked flask equipped with a magnetic stirrer was added Compound 10 (130mg, 0.497mmol), evacuated and replaced with N _2, were added dropwise under N ₂ atmosphere through a syringe of dry dichloromethane (2mL) and of DMF (2mL), The mixture was stirred and the mixture was cooled to 0 ° C, and oxalyl chloride (0.42 mL, 0.846 mmol, 2M in dichloromethane) was slowly added dropwise, and the mixture was dropped, and the ice bath was removed, and the reaction was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, added dry dichloromethane (10 mL), under N ₂ atmosphere clear solution was stirred and cooled to 0 deg.] C, was slowly added dropwise Compound 21 (122mg, 0.597mmol) in dichloromethane (2 mL), followed by the dropwise DIPEA (0.25 mL, 1.49 mmol) was added, the ice bath was removed, and the mixture was stirred at room temperature for 2 hours. The reaction was quenched by EtOAc (EtOAc)EtOAc. The yield was 35.1%. LC-MS (APCI): m / z = 453.4 (M + 1) + 1 H NMR (CDCl 3, 300MHz) (δ / ppm):. 9.06 (d, J = 12.0Hz, 1H), 8.41 (d, J = 6.9 Hz, 1H), 8.38 (s, 1H), 8.09 (d, J = 5.7 Hz, 2H), 7.94 (t, J = 6.0 Hz, 1H), 7.47 (s, 1H), 7.21 (d, J=9.3 Hz, 1H), 6.80 (s, 1H), 2.31 (s, 3H), 1.98-1.88 (m, 1H), 0.92-0.84 (m, 4H).

Biological activity test.

(1) Kinase inhibition

Reagents and consumables:

Eu-anti-GST抗体(Life Technologies,目录号PV5594)。 ASK1 (Invitrogen Cat. No. PV3809), ATP (Sigma, Cat. No. A7699), DMSO (Sigma, Cat. No. D8418-1L), 384-well plate (Greiner, Cat. No. 784076), HTRF KinEASE-STK Kit (Cisbio, Cat. No. PV3809), 5x Kinase Buffer A (Life Technologies, Cat. No. PV3186), Kinase Tracer 199 (Life Technologies, Cat. No. PV5830),

Eu-anti-GST antibody (Life Technologies, catalog number PV5594).

Specific experimental methods:

Compound formulation: The test compound was dissolved in DMSO to make a 20 mM mother liquor. Then, it was diluted 3 times in a medium gradient of DMSO and diluted 10 times. When dosing, dilute with buffer for 10 times.

ASK1 Kinase Assay: ASK1 kinase was mixed with pre-diluted compounds of different concentrations in 5x Kinase Buffer A for 10 minutes at each concentration. The corresponding substrate and ATP were added and reacted at room temperature for 20 minutes (in which a negative positive control was set: a negative blank control and a positive erlotinib). After the reaction, the detection reagent (reagent in the HTRF KinEASE-STK Kit) was added, and after incubation at room temperature for 30 minutes, the enzyme activity in the presence of the compound of the present invention at each concentration was measured by an Evnvision microplate reader, and different concentrations were calculated. The inhibitory activity of the compound on the enzyme activity was then fitted to the inhibition activity of the enzyme activity at different concentrations of the compound according to the four-parameter equation according to Graphpad 5.0 software, and the IC ₅₀ value was calculated.

The compound of the present invention and the undeuterated compound Selonsertib were tested in the above kinase inhibition assay, and the compounds of the present invention were found to have potent or comparable activity against ASK1 kinase. The results of inhibition of kinase by representative example compounds are summarized in Table 1 below.

Table 1

Example compound ASK1 IC ₅₀ (nM) Selonsertib 1.10 T-1 1.00 T-2 1.12 T-3 1.12 T-4 1.03

(2) Metabolic stability evaluation

Microsomal experiments: human liver microsomes: 0.5 mg/mL, Xenotech; rat liver microsomes: 0.5 mg/mL, Xenotech; mouse liver microsomes: 0.5 mg/mL, Xenotech; coenzyme (NADPH/NADH): 1 mM , Sigma Life Science; magnesium chloride: 5 mM, 100 mM phosphate buffer (pH 7.4).

Preparation of the stock solution: A certain amount of the powder of the example compound was accurately weighed and dissolved in 5 mM with DMSO.

Preparation of phosphate buffer (100 mM, pH 7.4): Mix 150 mL of 0.5 M potassium dihydrogen phosphate and 700 mL of 0.5 M potassium dihydrogen phosphate solution, and adjust the mixture with 0.5 M potassium dihydrogen phosphate solution. The pH of the solution was adjusted to 7.4, diluted 5 times with ultrapure water before use, and magnesium chloride was added to obtain a phosphate buffer (100 mM) containing 100 mM potassium phosphate, 3.3 mM magnesium chloride, and a pH of 7.4.

A solution of NADPH regeneration system (containing 6.5 mM NADP, 16.5 mM G-6-P, 3 U/mL G-6-P D, 3.3 mM magnesium chloride) was prepared and placed on wet ice before use.

Formulation stop solution: acetonitrile solution containing 50 ng/mL propranolol hydrochloride and 200 ng/mL tolbutamide (internal standard). Take 25057.5 μL of phosphate buffer (pH 7.4) into a 50 mL centrifuge tube, add 812.5 μL of human, rat and mouse liver microsomes, and mix to obtain a liver microsome dilution with a protein concentration of 0.625 mg/mL. . Incubation of the sample: The stock solution of the corresponding compound was diluted to 0.25 mM with an aqueous solution containing 70% acetonitrile as a working solution, and was used. 398 μL of human liver microsomes, rat liver microsomes or mouse liver microsome dilutions were added to a 96-well incubation plate (N=2), and 2 μL of 0.25 mM working solution was added and mixed.

Determination of metabolic stability: 300 μL of pre-cooled stop solution was added to each well of a 96-well deep well plate and placed on ice as a stop plate. The 96-well incubation plate and the NADPH regeneration system were placed in a 37 ° C water bath, shaken at 100 rpm, and pre-incubated for 5 min. 80 μL of the incubation solution was taken from each well of the incubation plate, added to the stopper plate, and mixed, and 20 μL of the NADPH regeneration system solution was added as a sample of 0 min. Then, 80 μL of the NADPH regeneration system solution was added to each well of the incubation plate to start the reaction and start timing. The corresponding compound had a reaction concentration of 1 μM and a protein concentration of 0.5 mg/mL. 100 μL of the reaction solution was taken at 10, 30, and 90 min, respectively, and added to the stopper, and the reaction was terminated by vortexing for 3 min. The plate was centrifuged at 5000 x g for 10 min at 4 °C. 100 μL of the supernatant was taken into a 96-well plate to which 100 μL of distilled water was previously added, mixed, and sample analysis was performed by LC-MS/MS.

Data analysis: The peak area of the corresponding compound and the internal standard was detected by LC-MS/MS system, and the ratio of the peak area of the compound to the internal standard was calculated. The slope is measured by the natural logarithm of the percentage of the remaining amount of the compound versus time, and t _1/2 and CL _{int are} calculated according to the following formula, where V/M is equal to 1/protein concentration.

The metabolic stability of liver microsomes in human, rat and mouse was evaluated by comparing the compound of the present invention with the undeuterated compound Selonsertib. The half-life and liver intrinsic clearance as indicators of metabolic stability are shown in Table 2. As shown in Table 2, in the human, rat and mouse liver microsome experiments, the compounds of the present invention significantly improved metabolic stability by comparison with the undeuterated compound Selonsertib.

Table 2

(3) Rat pharmacokinetic experiments

Six male Sprague-Dawley rats, aged 7-8 weeks, weighing approximately 210 g, were divided into two groups of 3 animals each, and their pharmacokinetic differences were compared by intravenous or oral single dose of compound (10 mg/kg orally). .

Rats were fed a standard diet and given water. Fasting began 16 hours before the test. The drug was dissolved with PEG400 and dimethyl sulfoxide. Blood was collected from the eyelids at a time point of 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, and 24 hours after administration.

Rats were briefly anesthetized after inhalation of ether, and 300 μL of blood samples were collected from the eyelids in test tubes. There was 30 μL of 1% heparin salt solution in the test tube. The tubes were dried overnight at 60 ° C before use. After the blood sample collection was completed at the last time point, the rats were anesthetized with ether and sacrificed.

Immediately after the blood sample is collected, gently invert the tube at least 5 times to ensure adequate mixing and place on ice. Blood samples were centrifuged at 5000 rpm for 5 minutes at 4 ° C to separate plasma from red blood cells. Pipette 100 μL of plasma into a clean plastic centrifuge tube, indicating the name and time of the compound. Plasma was stored at -80 °C prior to analysis. The concentration of the compound of the invention in plasma was determined by LC-MS/MS. Pharmacokinetic parameters were calculated based on the plasma concentration of each animal at different time points.

Experiments have shown that the compounds of the present invention have better pharmacokinetic properties in animals and therefore have better pharmacodynamics and therapeutic effects.

The above is a further detailed description of the present invention in connection with the specific preferred embodiments, and the specific embodiments of the present invention are not limited to the description. It will be apparent to those skilled in the art that the present invention may be made without departing from the spirit and scope of the invention.

Claims (14)

A 1,2,4-triazole compound of the formula (I), or a pharmaceutically acceptable salt thereof:

among them, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Y ¹ , Y ² , Y ³ , Y ⁴ , Y ⁵ , Y ⁶ are each independently hydrogen or hydrazine; X ¹ , X ² , X ³ are each independently CH ₃ , CH ₂ D, CHD ₂ or CD ₃ ; Provided that if X ¹ , X ² and X ^{3 are} each CH ₃ , then R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Y ¹ , Y ² , Y ³ , at least one of Y ⁴ , Y ⁵ and Y ⁶ is 氘. The 1,2,4-triazole compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ³ and R ⁴ are hydrazine. The 1,2,4-triazole compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R ⁵ is hydrazine. The 1,2,4-triazole compound according to any one of claims 1 to 3, wherein X ¹ and X ² are CD ₃ , or a pharmaceutically acceptable salt thereof. The 1,2,4-triazole compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, wherein Y ¹ is hydrazine. The 1,2,4-triazole compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

A pharmaceutical composition comprising a pharmaceutically acceptable excipient and the 1,2,4-triazole compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof. A process for the preparation of a pharmaceutical composition according to claim 7 comprising: a pharmaceutically acceptable excipient and the 1,2,4-triazole according to any one of claims 1-6 The compound or a pharmaceutically acceptable salt thereof is mixed to form a pharmaceutical composition. The 1,2,4-triazole compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 7 or 8 for use in treatment at least in part ASK1-mediated drug approach to disease. The method of claim 9 further comprising administering another therapeutic agent; preferably wherein said another therapeutic agent is a LOX2 inhibitor. The method according to claim 9 or 10, wherein the disease is diabetes, diabetic nephropathy, renal disease, renal fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), liver fibrosis, pulmonary hypertension, non-alcoholic Fatty liver hepatitis, liver disease, alcoholic liver disease, inflammatory disease, autoimmune disease, proliferative disease, transplant rejection, diseases involving impaired cartilage metabolism, congenital cartilage malformation, or diseases associated with excessive secretion of IL6 . The method of claim 11 wherein the disease is nonalcoholic steatohepatitis or alcoholic liver disease. The method of claim 11 wherein the disease is pulmonary hypertension or pulmonary fibrosis. The method of claim 11 wherein the disease is diabetic nephropathy, kidney disease or renal fibrosis.