CN109232538A

CN109232538A - A kind of 1,2,4- triazole compound

Info

Publication number: CN109232538A
Application number: CN201811159093.7A
Authority: CN
Inventors: 王义汉; 李焕银
Original assignee: Shenzhen Rui Rui Rui Biological Medicine Co Ltd
Current assignee: Shenzhen Rui Rui Rui Biological Medicine Co Ltd; Shenzhen Targetrx Inc
Priority date: 2017-10-12
Filing date: 2018-09-30
Publication date: 2019-01-18
Anticipated expiration: 2038-09-30
Also published as: CN111518080A; WO2019072130A1; CN109232538B

Abstract

The present invention relates to 1,2,4- triazole compound and its pharmaceutically acceptable salts shown in formula (I).The compound has Cell proliferation (" ASK1 ") inhibitor activity, therefore it can be used for treating the illness of ASK1 mediation, including chronic liver disease, cardiovascular disease, dysbolism, Respiratory Disturbances, gastrointestinal disorder and neurodegenerative disease.The present invention also provides the medical composition and its uses comprising the compounds of this invention.

Description

A kind of 1,2,4- triazole compound

Technical field

The invention belongs to pharmaceutical technology field, more particularly to a kind of 1,2,4- triazole compounds and include the compound Composition and application thereof.In particular it relates to which 5- (4- cyclopropyl -1H- imidazoles -1- base) -2- that certain deuteriums replace is fluoro- 4- methyl-N- [6- (4- isopropyl -4H-1,2,4,-triazole -3- base) -2- pyridine]-benzamide, the chemical combination that these deuteriums replace Object is used to treat the disease of ASK1 mediation, and has more excellent pharmacokinetic property.

Background technique

Many current drugs all suffer from absorption, distribution, metabolism and/or excretion (ADME) property of difference, and which prevent it Be applied even more extensively or limit them for certain indications.The ADME property of difference is also the drug candidate in clinical test The major reason of object failure.Such a problem is tachymetabolism, it causes many scripts that will highly have in disease treatment The drug of effect is excessively promptly removed from body.Quick medicament remove common solution be frequent or high dose administration with Reach sufficiently high Plasma Drug Level.However, which results in many potential treatment problems, such as patient is to therapeutic regimen Compliance is poor, side effect becomes more violent and increases treatment cost under higher dosage.Rapid drug is metabolized to be also possible to make to suffer from Person is exposed to undesirable toxicity or reactive metabolin.

Another ADME limitation for influencing many drugs is to form toxicity or biological reactivity metabolin.Then, receive Some patients of the drug, which can suffer from toxicity, or such drug safe dose may be restricted causes patient to receive The activating agent of suboptimal amount.In some cases, change dosing interval or formulation method potentially contribute to reduce clinical bad anti- It answers, but being generally formed in the metabolism of the compound for this undesirable metabolin is intrinsic.

A kind of strategy for having potential attraction for improving drug metabolism performance is deuterium modification.In this approach, Ren Menshi Figure slows down drug metabolism by substituting one or more hydrogen atoms with D-atom or reduces the formation of undesirable metabolin.Deuterium It is safe and stable, on-radiation hydrogen isotope.Compared with hydrogen, deuterium and carbon form stronger key.In the case where selection, by Deuterium assign bond strength increase can with the ADME property of positive influence drug, thus generate improve efficacy of drugs, safety and/or The potentiality of tolerance.Meanwhile because the size and shape of deuterium and the size and shape of hydrogen are essentially identical, with only include hydrogen it is original Chemical entities compare, by the estimated biochemical activity and selectivity that will not affect that drug of deuterium substitution hydrogen.

Between past 35 years, the influence (ginseng to metabolic rate is replaced to the approval drug report of considerably less ratio deuterium See, such as Foster, AB, Adv Drug Res, 1985,14:1-40 (" Foster ")；Fisher, MB etc., Curr Opin Drug Discov Devel, 2006,9:101-09 (" Fisher)).The result is that changeable and unpredictable.To some chemical combination Object, deuterate cause internal metabolic clearance rate to reduce.To other compounds, without metabolic alterations.Compound also shows generation Thank to the increase of clearance rate.The mutability of deuterium effect also causes technical staff to suspect or abandon deuterium modification as the unfavorable metabolism of inhibitor Feasible drug design strategies (page 101 referring to page 35 of Foster and Fisher).

Influence of the deuterium modification to pharmacokinetic properties is not predictable, even if penetrating into known metabolism site in D-atom In the case where.People determine whether and how metabolic rate is different from only possible through the practical drug for preparing and testing deuterate The counterpart of its non-deuterate.Many drug tools are likely to occur multiple sites of metabolism.It needs to carry out the substituted site of deuterium and sees To the influence (if any) to metabolism, necessary degree of deuterium will be different for various drugs.

The present invention relates to the new derivatives of Selonsertib and its pharmaceutically acceptable salts.The present invention also provides Composition comprising the compounds of this invention and such composition are passing through application ASK1 (Cell proliferation) suppression Preparation obtains the purposes in the disease of advantageous treatment and the treatment method of illness.

Selonsertib also known as GS-4997 and entitled 5- (4- cyclopropyl -1H- imidazoles -1- base) the fluoro- 4- of -2- of chemistry Methyl-N- [6- (4- isopropyl -4H-1,2,4,-triazole -3- base) -2- pyridine]-benzamide (has knot as follows Structure), it is the micromolecular inhibitor of lucky moral drugmaker research and development, can effectively reduce the pathologic function of ASK1.Selonsertib exists Treat nonalcoholic fatty liver disease (NASH) clinic second phase the experimental results showed that, only just played in NASH patient after 24 weeks Anti-fibrosis effect.Currently, the research that Selonsertib is used to treat NASH is in clinical three phases, for treating Alcoholic The research of hepatitis is in the clinical second phase.

The phosphorylation of ASK1 albumen can lead to apoptosis or other cellular responses according to cell type.ASK1 activation and signal Conduction be reported in including being played an important role in following broad range of diseases: neurodegeneration obstacle, cardiovascular disorder, Inflammatory disorder and dysbolism.In addition, ASK1 is involved in after ischaemic and the Reperfu- sion of mediate cardiac, brain and kidney Organ damage (Watanabe et al. (2005) BBRC 333,562-567；Zhang et al. (2003) LifeSci74-37-43； Terada et al. (2007) BBRC364:1043-49).

Even if existing Selonsertib, there are still have the medicine improved in the treatment of ASK1 activation related disease For the needs for the active compound that dynamics and/or pharmacodynamics show.

Summary of the invention

Against the above technical problems, the invention discloses a kind of 1,2,4- triazole compounds and include the compound Composition and application thereof, the compound have stronger ASK1 inhibitor activity.

In this regard, the invention adopts the following technical scheme:

The first aspect of the present invention is related to a kind of 1,2,4- triazole compounds of formula (I) or its is pharmaceutically acceptable Salt:

Wherein,

R¹、R²、R³、R⁴、R⁵、R⁶、R⁷、R⁸、Y¹、Y²、Y³、Y⁴、Y⁵And Y⁶It is from hydrogen or deuterium each independently；

X¹、X²And X³It is CH each independently₃、CH₂D、CHD₂Or CD₃。

Condition is if X¹、X²And X³Each is CH₃, then R¹、R²、R³、R⁴、R⁵、R⁶、R⁷、R⁸、Y¹、Y²、Y³、Y⁴、Y⁵With Y⁶At least one of be deuterium.

As the preferred embodiments of the invention, compound at least contains a D-atom, more preferably a deuterium in formula (I) Atom, more preferably two D-atoms, more preferably three D-atoms, more preferably four D-atoms, more preferably six D-atoms, more preferably Seven, ground D-atom, more preferably nine D-atoms.

As the preferred embodiments of the invention, it is same that deuterium isotopic content of the deuterium in deuterated position is at least greater than natural deuterium Position cellulose content 0.015%, is preferably greater than 30%, even more preferably greater than 50%, even more preferably greater than 75%, even more preferably greater than 95%, Even more preferably greater than 99%.

Specifically, R in the present invention¹、R²、R³、R⁴、R⁵、R⁶、R⁷、R⁸、Y¹、Y²、Y³、Y⁴、Y⁵、Y⁶、X¹、X²And X³, each deuterium Subrogate and set middle deuterium isotopic content and be at least 5%, is preferably greater than 10%, even more preferably greater than 15%, even more preferably greater than 20%, more It is greater than 25%, even more preferably greater than 30%, even more preferably greater than 35%, even more preferably greater than 40%, even more preferably greater than 45% goodly, more preferably Ground is greater than 50%, even more preferably greater than 55%, even more preferably greater than 60%, even more preferably greater than 65%, even more preferably greater than 70%, more preferably Greater than 75%, even more preferably greater than 80%, even more preferably greater than 85%, even more preferably greater than 90%, even more preferably greater than 95%, more preferably greatly In 99%.

In another embodiment, in formula (I) compound R¹、R²、R³、R⁴、R⁵、R⁶、R⁷、R⁸、Y¹、Y²、Y³、Y⁴、 Y⁵、Y⁶、X¹、X²And X³, at least one of which is containing deuterium, and more preferably two contain deuterium, and more preferably three contain deuterium, and more preferably four contain deuterium, More preferably five contain deuterium, and more preferably six contain deuterium, and more preferably seven contain deuterium, and more preferably eight contain deuterium, and more preferably nine contain deuterium, more It is good ground ten contain deuterium, more preferably 11 contain deuterium, more preferably 12 contain deuterium, more preferably 13 contain deuterium, more preferably 14 Containing deuterium, more preferably 15 contain deuterium, and more preferably 16 contain deuterium, and more preferably 17 contain deuterium, and more preferably 18 contain deuterium, more preferably 19, ground contains deuterium, and more preferably 20 contain deuterium, and more preferably 21 contain deuterium containing deuterium, more preferably 22, and more preferably two 13 contain deuterium.Specifically, in formula (I) compound at least contain one, two, three, four, five, six, seven, Eight, nine, ten, 11,12,13,14,15,16,17,18,19 A, 20,21,22,23 D-atoms.

As the preferred embodiments of the invention, R¹、R²、R³、R⁴、R⁵、R⁶、R⁷And R⁸It is from hydrogen or deuterium each independently.

In another preferred embodiment of the present, R¹、R³And R⁴It is deuterium.

In another preferred embodiment of the present, R⁵It is deuterium.

In another preferred embodiment of the present, R¹、R³、R⁴And R⁵It is deuterium.

As the preferred embodiments of the invention, Y¹、Y²、Y³、Y⁴、Y⁵And Y⁶It is from hydrogen or deuterium each independently.

In another preferred embodiment of the present, Y¹It is deuterium.

In another preferred embodiment of the present, Y²、Y³、Y⁴、Y⁵And Y⁶It is deuterium.

As the preferred embodiments of the invention, X¹、X²And X³Independent is CH₃、CH₂D、CHD₂Or CD₃。

In another preferred embodiment of the present, X¹、X²It is CD₃。

In another preferred embodiment of the present, X³It is CD₃。

As in the preferred embodiments of the invention, the compound is selected from the group compound or its is pharmaceutically acceptable Salt:

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

The second aspect of the present invention, the invention also discloses a kind of pharmaceutical compositions, contain pharmaceutically acceptable tax Shape agent and 1,2,4- triazole compound as described above or its pharmaceutically acceptable salt.

The third aspect of the present invention, the invention also discloses a kind of preparation method of pharmaceutical composition as described above, packets Include following steps: by pharmaceutically acceptable excipient and as described above 1,2,4- triazole compounds or its pharmaceutically Acceptable salt is mixed, to form pharmaceutical composition.

In another preferred example, described pharmaceutical composition is injection, wafer, tablet, pill, powder or granule.

In another preferred example, the pharmaceutical composition also contains other therapeutic agent, the other medicine Object is treating cancer, cardiovascular disease, inflammation, infection, immunity disease, metabolic disease or the drug of organ transplant.

The fourth aspect of the present invention additionally provides a kind of disease treated in the patient of needs and at least partly mediated by ASK1 Disease method, the compound of the first aspect present invention of patient's treatment effective dose including giving needs or its can pharmaceutically connect The salt received or its pharmaceutical composition.

In specific embodiments, the medicable disease of the compounds of this invention be selected from diabetes, diabetic nephropathy, nephrosis, Kidney fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), liver fibrosis, pulmonary hypertension, nonalcoholic fatty liver hepatitis, Hepatopathy, alcoholic liver disease, inflammatory conditions, autoimmune disease, proliferative diseases, graft rejection, to be related to cartilage metabolism impaired Disease, congenital cartilage deformity or relevant to IL6 hypersecretion disease.

In another preferred example, medicable disease is nonalcoholic steatohepatitis or alcoholic liver disease.

In another preferred example, medicable disease is pulmonary hypertension or pulmonary fibrosis.

In another preferred example, medicable disease is diabetic nephropathy, nephrosis or kidney fibrosis.

It should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the invention, embodiment and (such as implementation below Example) in specifically describe each technical characteristic between can be combined with each other, to form a new or preferred technical solution.It is limited to Length, not repeated them here.

Specific embodiment

Compound and its pharmaceutically acceptable salt and the not deuterated compound phase of deuterated ASK1 inhibitor of the invention Than there is more preferably pharmacokinetics and/or pharmacodynamics performance, therefore be more suitable for the compound of ASK1 inhibitor, in turn The drug for the related disease that more applicable preparation treatment ASK1 is mediated.The present invention is completed on this basis.

Definition

Herein, unless otherwise instructed, " deuterated " refers to one or more hydrogen in compound or group replaced deuterium；Deuterium In generation, can be a substitution, two replace, polysubstituted or full substitution.Term " one or more deuterated " and " one or many deuterated " It is used interchangeably.

Herein, unless otherwise instructed, " non-deuterated compound " refers to ratio containing D-atom not higher than the same position of natural deuterium The compound of cellulose content (0.015%).

The invention also includes the compounds of isotope labelling, are equal to original chemical and are disclosed.This hair can be classified as The example of bright compound isotope includes hydrogen, carbon, nitrogen, oxygen, phosphorus, sulphur, fluorine and chlorine isotope, respectively such as²H,³H,¹³C,¹⁴C,¹⁵N,¹⁷O,¹⁸O,³¹P,³²P,³⁵S,¹⁸F and³⁶Cl.Compound or enantiomer in the present invention, diastereomer, isomers or medicine Acceptable salt or solvate on, wherein containing the isotope of above compound or other other isotope atoms all at this Within the scope of invention.Certain compound isotopically labelleds in the present invention, such as³H and¹⁴The radioactive isotope of C also wherein, It is useful in the experiment of the Tissue distribution of drug and substrate.Tritium, i.e.,³H and carbon-14, i.e.,¹⁴C, their preparation and detection are compared It is easy, is the first choice in isotope.The compound of isotope labelling can use general method, by with the isotope mark being easy to get Note reagent replaces with non isotopic reagent, can be prepared with the scheme in example.

Compound

The present invention provides formula (I) compound or its pharmaceutically acceptable salt, prodrug, hydrate or solvated compounds, crystalline substance Type, stereoisomer or isotopic variations:

Wherein,

R¹、R²、R³、R⁴、R⁵、R⁶、R⁷、R⁸、Y¹、Y²、Y³、Y⁴、Y⁵、Y⁶It is from hydrogen or deuterium each independently；

X¹、X²、X³It is CH each independently₃、CH₂D、CHD₂Or CD₃；

As the preferred embodiments of the invention, deuterium isotopic content of the deuterium in deuterated position is at least greater than natural same position Cellulose content 0.015% is preferably greater than 30%, and more preferable moral is greater than 50%, more preferably greater than 75%, more preferably greater than 95%, more preferably greater than 99%.

In the specific embodiment of logical formula (I), " R¹、R²、R³、R⁴、R⁵、R⁶、R⁷And R⁸It is from hydrogen or deuterium each independently " Including R¹Selected from hydrogen or deuterium, R²Selected from hydrogen or deuterium, R³Selected from hydrogen or deuterium, and so on, until R⁸Technical side selected from hydrogen or deuterium Case.More specifically, including R¹It is hydrogen or R¹It is deuterium, R²It is hydrogen or R²It is deuterium, R³It is hydrogen or R³It is deuterium, and so on, until R⁸It is Hydrogen or R⁸It is the technical solution of deuterium.

In another specific embodiment of logical formula (I), " Y¹、Y²、Y³、Y⁴、Y⁵And Y⁶It is from hydrogen or deuterium each independently " Including Y¹Selected from hydrogen or deuterium, Y²Selected from hydrogen or deuterium, Y³Selected from hydrogen or deuterium, and so on, until Y⁶Technical side selected from hydrogen or deuterium Case.More specifically, including Y¹It is hydrogen or Y¹It is deuterium, Y²It is hydrogen or Y²It is deuterium, Y³It is hydrogen or Y³It is deuterium, and so on, until Y⁶It is Hydrogen or Y⁶It is the technical solution of deuterium.

In another specific embodiment of logical formula (I), " X¹、X²、X³It is CH each independently₃、CH₂D、CHD₂Or CD₃” Including X¹Selected from CH₃、CH₂D、CHD₂Or CD₃, X²Selected from CH₃、CH₂D、CHD₂Or CD₃, X³Selected from CH₃、CH₂D、CHD₂Or CD₃Skill Art scheme.More specifically, including X¹It is CH₃、X¹It is CH₂D、X¹It is CHD₂Or X¹It is CD₃, X²It is CH₃、X²It is CH₂D、X²It is CHD₂Or X²It is CD₃, X³It is CH₃、X³It is CH₂D、X³It is CHD₂Or X³It is CD₃Technical solution.

In being preferably carried out scheme, the present invention relates to formula (I) compound or its pharmaceutically acceptable salt, prodrug, water Close object or solvated compounds, crystal form, stereoisomer or isotopic variations, wherein R⁷And R⁸、Y²-Y⁶It is hydrogen, R¹-R⁸And Y¹Respectively From being independently hydrogen or deuterium, X¹、X²And X³It is CH each independently₃、CH₂D、CHD₂Or CD₃, additional conditions be the compound extremely Contain a D-atom less.

In a further preferred embodiment, R²And R⁶It is hydrogen.

In a further preferred embodiment, R¹、R³And R⁴It is each independently selected from hydrogen or deuterium.Another more preferably real It applies in scheme, R¹、R³And R⁴It is hydrogen.In another more preferably embodiment, R¹、R³And R⁴It is deuterium.

In a further preferred embodiment, R⁵It is hydrogen or deuterium.In another more preferably embodiment, R⁵It is hydrogen.Another One more preferably in embodiment, R⁵It is deuterium.

In a further preferred embodiment, X³It is CH₃。

In a further preferred embodiment, X¹And X²It is each independently selected from CH₃Or CD₃.More preferably implement another In scheme, X¹And X²It is CH₃.In another more preferably embodiment, X¹And X²It is CD₃。

In a further preferred embodiment, Y¹It is hydrogen or deuterium.In another more preferably embodiment, Y¹It is hydrogen.Another One more preferably in embodiment, Y¹It is deuterium.

Wherein, term " pharmaceutically acceptable salt " refers to, in reliable medical judgment scope, is suitble to and people and low The tissue of animal contacts without excessive toxicity, irritation, allergy etc., and with reasonable benefit/hazard ratio phase Those of title salt.Pharmaceutically acceptable salt is well known in the art.For example, Berge et al. exists The pharmaceutically acceptable salt being described in detail in J.Pharmaceutical Sciences (1977) 66:1-19.Chemical combination of the present invention The pharmaceutically acceptable salt of object includes derived from suitable inorganic and organic acid and inorganic and organic base salt.It can pharmaceutically connect The example for the nontoxic acid-addition salts received is the salt formed with inorganic acid, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, Or the salt formed with organic acid, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid.It also include making The salt formed with conventional method in that art, for example, ion-exchange process.Other pharmaceutically acceptable salts include: hexanedioic acid salt, Alginate, ascorbate, aspartate, benzene sulfonate, benzoate, bisulphate, borate, butyrate, camphor Hydrochlorate, camsilate, citrate, cipionate, digluconate, lauryl sulfate, esilate, formic acid Salt, fumarate, gluconate, glycerophosphate, gluconate, Hemisulphate, enanthate, caproate, hydriodate, 2- Hydroxy-ethanesulfonate salt, Lactobionate, lactate, laruate, lauryl sulfate, malate, maleate, malonic acid Salt, mesylate, 2- naphthalene sulfonate, nicotinate, nitrate, oleate, oxalates, palmitate, embonate, pectin ester Hydrochlorate, persulfate, 3- phenpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulphur Hydrochlorate, tartrate, rhodanate, tosilate, undecanoate, valerate, etc..Medicine derived from suitable alkali Acceptable salt includes alkali metal, alkaline-earth metal, ammonium and N on⁺(C_1-4Alkyl)₄Salt.Representative alkali or alkaline earth metal Salt includes sodium, lithium, potassium, calcium, magnesium salts, etc..If applicable, other pharmaceutically acceptable salts include and counter ion shape At nontoxic ammonium salt, quaternary ammonium salt and amine cation, counter ion for example halogen ion, hydroxyl, carboxylate radical, sulfate radical, phosphate radical, Nitrate anion, loweralkyl sulfonate and arylsulphonate.

Term " solvate " refers to that the compounds of this invention and solvent molecule are coordinated the complex to form special ratios." hydration Object " refers to that the compounds of this invention and water carry out the complex of coordination formation.

Term " prodrug " includes that itself can be biologically active or inactive, is taken when with method appropriate Afterwards, it is metabolized or is chemically reacted in human body and change a kind of compound of an accepted way of doing sth (I) or a compound of formula (I) Composed salt or solution.The prodrug includes but is not limited to carboxylate, carbonic ester, phosphate, the nitre of said compound Acid esters, sulfuric ester, sulfone ester, sulfoxide esters, amino-compound, carbaminate, azo-compound, phosphamide, glucoside, ether, The forms such as acetal.

Pharmaceutical composition and method of administration

Since the compounds of this invention has an activity of excellent inhibition ASK1 kinases, the compounds of this invention and its various Crystal form, pharmaceutically acceptable salt, hydrate or solvate, and containing the compounds of this invention be main active medicine Compositions can be used for treating, prevent and alleviate the disease that ASK1 is mediated.According to the prior art, the compounds of this invention can be used for Treat following disease: chronic liver disease, cardiovascular disease, dysbolism, Respiratory Disturbances, gastrointestinal disorder and neurodegenerative disease Deng.

Pharmaceutical composition of the invention include safe and effective amount within the scope of the compounds of this invention or its be pharmacologically subjected to Salt and pharmacologically acceptable excipient or carrier.Wherein " safe and effective amount " refers to: the amount of compound is enough obviously Improve the state of an illness, and is unlikely to generate serious side effect.In general, pharmaceutical composition contain 0.5-2000mg the compounds of this invention/ Agent more preferably contains 1-500mg the compounds of this invention/agent.Preferably, described is " one " for a capsule or tablet.

" pharmaceutically acceptable excipient " refers to the nontoxic of the pharmacological activity that will not destroy the compound deployed together Carrier, adjuvant or mediator.Can be used for pharmaceutically acceptable carrier, adjuvant or mediator in the present composition include (but It is not limited to) ion-exchanger, aluminium oxide, aluminum stearate, lecithin, haemocyanin (such as human serum albumin), buffer substance (such as phosphate), glycine, sorbic acid, potassium sorbate, the partial glyceride mixture of saturated vegetable fatty acid, water, salt or electrolysis Matter (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, silica gel, magnesium trisilicate, polyvinylpyrrolidine Ketone, the substance based on cellulose, polyethylene glycol, sodium carboxymethylcellulose, polyacrylate, wax, polyethylene-polyoxypropylene-are embedding Section polymer, polyethylene glycol and lanolin.

The method of application of the compounds of this invention or pharmaceutical composition is not particularly limited, and representative method of application includes (but being not limited to): oral, duodenum, rectum, parenteral (intravenous, intramuscular or subcutaneous) and local administration.

Solid dosage forms for oral administration includes capsule, tablet, pill and granule.In these solid dosage forms, Reactive compound is mixed at least one conventional inert excipients (or carrier), such as sodium citrate or Dicalcium Phosphate, or with it is following Ingredient mixing: (a) filler or solubilizer, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid；(b) adhesive, example Such as, hydroxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and Arabic gum；(c) moisturizer, for example, sweet Oil；(d) disintegrating agent, for example, agar, calcium carbonate, potato starch or tapioca, alginic acid, certain composition silicates and carbonic acid Sodium；(e) retarding solvent, for example, paraffin；(f) absorbsion accelerator, for example, quaternary ammonium compound；(g) wetting agent, for example, cetanol and Glycerin monostearate；(h) adsorbent, for example, kaolin；(i) lubricant, for example, talcum, calcium stearate, magnesium stearate, Or mixtures thereof solid polyethylene glycol, lauryl sodium sulfate,.In capsule, tablet and pill, dosage form also may include buffering Agent.

Coating and shell material preparation can be used in solid dosage forms such as tablet, sugar-pill, capsule, pill and granule, such as casing and Other materials well known in the art.They may include opacifying agent, also, reactive compound or compound in this composition Release can discharge in certain a part in the digestive tract in a delayed fashion.The example of adoptable embedding component is polymeric material And wax material.When necessary, reactive compound can also be with one of above-mentioned excipient or a variety of formation microencapsulation forms.

Liquid formulation for oral administration includes pharmaceutically acceptable lotion, solution, suspension, syrup or tincture. In addition to active compounds, liquid dosage form may include the inertia releasing agent routinely used in this field, such as water or other solvents, increase Solvent and emulsifier, for example, ethyl alcohol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-BDO, dimethyl formyl The mixing of amine and oil, especially cottonseed oil, peanut oil, maize germ, olive oil, castor oil and sesame oil or these substances Object.

Other than these inert diluents, composition also may include auxiliary agent, such as wetting agent, emulsifier and suspending agent, sweet taste Agent, corrigent and fragrance.

In addition to active compounds, suspension may include suspending agent, for example, ethoxylation isooctadecane alcohol, polyoxyethylene Sorbierite and anhydro sorbitol, microcrystalline cellulose, aluminium methoxide and agar or the mixture of these substances etc..

Composition for parenteral injection may include physiologically acceptable sterile, aqueous or anhydrous solution, dispersion liquid, Suspension or lotion, and the aseptic powdery for re-dissolving into sterile Injectable solution or dispersion liquid.It is suitable aqueous and Nonaqueous carrier, diluent, solvent or excipient include water, ethyl alcohol, polyalcohol and its suitable mixture.

The dosage form of the compounds of this invention for local administration includes ointment, powder, patch, stock solution and inhalant. Active constituent aseptically with physiologically acceptable carrier and any preservative, buffer, or when necessary may need Propellant be mixed together.

The compounds of this invention can be administered alone, or be administered in combination with other pharmaceutically acceptable compounds.

It is the mammal that the compounds of this invention of safe and effective amount is applicable to treatment when using pharmaceutical composition (such as people), wherein dosage is the effective dosage pharmaceutically thought when application, for the people of 60kg weight, day is to medicament Amount is usually 0.5~2000mg, preferably 1~500mg.Certainly, specific dosage is also contemplated that administration route, patient health situation etc. Factor, within the scope of these are all skilled practitioners technical ability.

The method for treating disease

Therapeutic agent as ASK1 signal transduction inhibitor has following potentiality: treatment needs to treat the trouble of disease or illness Neurodegenerative disease, cardiovascular disease, inflammation, autoimmune disease and the dysbolism of person improve its life.In particular, ASK1 inhibitor has following potentiality: treatment cardiorenal disease, including kidney trouble, nephrosis, chronic kidney disease, fibrosis Disease (including lung and kidney fibrosis), dilated cardiomyopathy, respiratory disease (including chronic obstructive pulmonary disease (COPD) and anxious Property injury of lungs), acute and chronic hepatitis (such as nonalcoholic steatohepatitis and alcoholic hepatitis).

For authenticating compound inhibit ASK1 kinase activity ability and its validity as ASK1 inhibitor it is a variety of Test is known in the art and is described in, for example, U.S. Patent number 8,742,126.

Some embodiments as described herein be related to compound as described herein form or pharmaceutical composition as described herein Object treatment needs the purposes of the disease of the patient with ASK1 inhibitor for treating.

Some embodiments as described herein are the method for treating diabetic nephropathy or diabetic complication, including administration is controlled Treat the form or pharmaceutical composition as described herein of a effective amount of compound formula (I) as described herein.In some embodiments, Diabetes include 1 type and diabetes B, gestational diabetes mellitus, prediabetes, insulin resistance, metabolic syndrome, fasting blood-glucose Impaired and impaired glucose tolerance.Type 1 diabetes are also referred to as insulin-dependent diabetes mellitus (IDDM).2 types are also referred to as non-pancreas islet Element-dependent diabetes (NIDDM).

Another embodiment is related to the method for treating nephrosis or nephrosis, this paper including dosage treatment effective amount The form or pharmaceutical composition as described herein of the compound formula (I).

Another embodiment is related to the method for treating kidney fibrosis, pulmonary fibrosis or idiopathic pulmonary fibrosis (IPF), including The form or pharmaceutical composition as described herein of the compound I as described herein of dosage treatment effective amount.

Another embodiment is related to treatment of diabetic nephropathy, diabetic nephropathy, kidney fibrosis, liver fibrosis or lung fiber The method of change, the crystal form or pharmaceutical composition as described herein of the compound I as described herein including dosage treatment effective amount Object.

Disclosed herein is the methods for the treatment of and/or prevention of liver disease in the patient of needs, including have to patient's drug treatment The form of compound I as described herein of effect amount or combinations thereof object is optionally combined with the LOXL2 inhibitor of therapeutically effective amount.It is living The presence of dynamic property hepatopathy can be detected by the presence of enzyme level raised in blood.Specifically, it is known that be higher than clinically The alanine aminotransferase (ALT) of the normal range (NR) of receiving and the blood level of aspartate transaminase (AST) indicate The hepatic injury of progress.Measurement is used clinically for the routine monitoring of the blood level of the ALT and AST of hepatopath to control in medical treatment The progress of hepatopathy when treatment.Raised ALT and AST are reduced in the normal range (NR) of receiving as clinical evidence, reflection patient is just In the reduction of the seriousness of the hepatic injury of progress.

In certain embodiments, hepatopathy is chronic liver disease.Chronic liver disease, which is related to hepatic parenchymal progressive, to be destroyed and regenerates, Lead to fibrosis and cirrhosis.In general, chronic liver disease can be by virus (such as hepatitis B, hepatitis C, cytomegalovirus (CMV) or Epstein Barr is viral (EBV)), toxic agents or drug (such as alcohol, methotrexate (MTX) or furantoin), metabolism Property disease (such as non-alcohol fatty liver (NAFLD), nonalcoholic fatty liver disease (NASH), hemochromatosis or Wilson's disease), autoimmune disease (such as autoimmune chronic hepatitis, primary biliary cirrhosis or primary Sclerosing cholangitis) or other reasons (such as right heart failure) cause.

In a particular embodiment, the hepatopathy is Metabolic liver disease.In one embodiment, the hepatopathy is non-wine Essence fatty liver disease (NAFLD).NAFLD and insulin resistance and metabolic syndrome (obesity, combined hyperlipidemia familial, sugar Urinate sick (II type) and hypertension) it is related.Think that NAFLD covers a series of disease activities, and starts as the accumulation of fat in liver (hepatic steatosis).

The compound of the present invention has series of advantages compared with non-deuterated compound well known in the prior art.This hair Bright advantage includes: first, has excellent inhibition to ASK1 protein kinase using the compound of technical solution of the present invention.The Two, metabolism of the compound in organism is improved, makes compound that there is better pharmacokinetic parameter characteristic.In this feelings Under condition, thus it is possible to vary dosage simultaneously forms durative action preparation, improves applicability.Third improves the drug of compound in animal body Concentration improves curative effect of medication.4th, it is suppressed that certain metabolites improve the safety of compound.

Embodiment

Combined with specific embodiments below, it is further elaborated the present invention.It should be understood that these embodiments are merely to illustrate this hair It is bright rather than limit the scope of the invention.In the following examples, the experimental methods for specific conditions are not specified, usually according to routine Condition, or according to the normal condition proposed by manufacturer.Unless otherwise stated, otherwise parts and percentages are weight proportion and weight hundred Divide ratio.

In general, each reaction is usually in atent solvent, in room temperature to reflux temperature (such as 0 DEG C~100 in preparation flow DEG C, preferably 0 DEG C~80 DEG C) under carry out.Reaction time is usually 0.1-60 hours, it is therefore preferable to 0.5-24 hours.

1 5- of embodiment (4- cyclopropyl -1H- imidazole radicals -1- base)-N- (6- (4- isopropyl -4H-1,2,4- triazole -3- Base -5-d) pyridine -2- base -3,4-d₂) the fluoro- 4- methyl benzamide (compound T-1) of -2- preparation.

Specific synthesis step is as follows:

The synthesis of step 1 compound 2.

Compound 1 (5.0g, 32.86mmol) and methanol are successively added into the 100mL single-necked flask equipped with magnetic agitation (60mL) stirs dissolved clarification, is slowly added dropwise into hydrazine hydrate (3.29g, 65.72mmol), after dripping off, reaction mixture is heated at reflux 3 Hour, it is then cooled to room temperature, a large amount of white solids is precipitated, filter, the washing of filter cake cold methanol, dry the white solid 3.5g, yield 70%.LC-MS (APCI): m/z=153.2 (M+1)⁺.¹H NMR(DMSO-d₆,300MHz)(δ/ppm):9.14 (s, 1H), 7.51 (t, J=5.7Hz, 1H), 7.11 (d, J=5.7Hz, 1H), 6.61 (d, J=6.0Hz, 1H), 6.08 (s, 2H),4.48(s,2H).

The synthesis of step 2 compound 3.

Successively toward equipped with magnetic agitation 100mL single-necked flask in be added compound 2 (3.5g, 23mmol), toluene (Tol, 50mL), isopropylamine (9.8g, 166mmol) and n,N-Dimethylformamide-dipropyl acetal (DMF-DPA, 10.89g, 62mmol), it is slowly added dropwise into acetic acid (2.1g, 35mmol), finishes, N under stirring₂Reaction mixture is heated at reflux for 24 hours under atmosphere.It is cold But room temperature is arrived, evaporating solvent under reduced pressure is added water (40mL), stirs the lower a large amount of solids of precipitation, filtering, filter cake isopropanol washs white Color solid 3.1g, yield 66.3%.LC-MS (APCI): m/z=204.2 (M+1)⁺.¹H NMR(CDCl₃,300MHz)(δ/ ppm):8.31(s,1H),7.58-7.54(m,2H),6.56(dd,J₁=5.4Hz, J₂=1.5Hz, 1H), 5.64-5.57 (m, 1H), 4.45 (s, 2H), 1.52 (d, J=4.8Hz, 6H)

The synthesis of step 3 compound 4.

Successively toward equipped with magnetic agitation 50mL tube sealing in be added compound 3 (200mg, 984umol), heavy water (10mL) and Pd/C (50mg, 10%), logical hydrogen is bubbled 5 minutes under stirring, 110 DEG C is warming up to after closed, and insulated and stirred reaction is overnight.It is cold But room temperature is arrived, is added methylene chloride (20mL), is filtered, layering, water phase methylene chloride extracts (20mLx2), merges organic phase, nothing Aqueous sodium persulfate dries, filters, and concentration obtains white solid 120mg, yield 59.1% through silica gel post separation.LC-MS(APCI):m/z =207.3 (M+1)⁺.¹H NMR(CDCl₃,300MHz)(δ/ppm):7.56(s,1H),5.62-5.59(m,1H),4.50(s, 1H), 1.51 (d, J=5.4Hz, 6H)

The synthesis of step 4 compound 7.

Compound 6 (5.0g, 24.5mmol), toluene are successively added into the 100mL single-necked flask equipped with magnetic agitation (50mL) stirs dissolved clarification, be added dropwise to compound 5 (4.4g, 27.0mmol) and n,N-diisopropylethylamine (DIPEA, 8.5mL, 51.5mmol), reaction solution is heated at reflux 2 hours.It is cooled to room temperature, is added water (50mL), layering, organic phase is successively with saturation NH₄Cl aqueous (20mL), saturation NaHCO₃Aqueous (20mL) and saturated salt solution (20mL) washing, anhydrous sodium sulfate is dry, mistake Filter, concentration, residue recrystallize in n-hexane, filter, dry to obtain off-white powder 3.1g, yield 44.1%.LC-MS (APCI): m/z=286.2&288.2 (M+1)⁺.¹H NMR(DMSO-d₆, 300MHz) and (δ/ppm): 7.07 (d, J=6.9Hz, 1H), 6.52 (d, J=4.5Hz, 1H), 5.29 (t, J=4.2Hz, 1H), 4.18 (d, J=4.2Hz, 2H), 4.52-4.51 (m, 1H),2.11(s,3H),0.98-0.88(m,4H).

The synthesis of step 5 compound 8.

Compound 7 (3.1g, 10.8mmol) and dichloromethane are successively added into the 50mL single-necked flask equipped with magnetic agitation Alkane (DCM, 15mL) stirs dissolved clarification and is cooled to 0 DEG C, is slowly added dropwise into formic acid (15mL) and acetic anhydride (Ac₂O, 4.1mL, 43.3mmol), reaction solution N₂It is stirred to react at 0 DEG C 3 hours under atmosphere.Water (20mL) quenching reaction, 40%NaOH aqueous is added PH to 9 to be adjusted, organic phase is isolated, water phase methylene chloride extracts (30mLx2), merges organic phase, and anhydrous sodium sulfate dries, filters, Concentrate is directly used in react in next step.LC-MS (APCI): m/z=314.1&316.1 (M+1)⁺.¹H NMR(DMSO-d₆, 300MHz) (δ/ppm): 8.15 (d, J=9.6Hz, 1H), 7.61 (d, J=5.4Hz, 1H), 7.41 (d, J=7.2Hz, 1H), 4.68(d,2H),2.50(s,3H),2.12-1.98(m,1H),0.96-0.85(m,4H).

The synthesis of step 6 compound 9.

Compound 8 (3.39g, 10.8mmol) and glacial acetic acid are successively added into the 50mL single-necked flask equipped with magnetic agitation (30mL) stirs dissolved clarification, is added ammonium acetate (2.57g, 42.1mmol), N₂Reaction mixture heated overnight at reflux under atmosphere.It is cooling To room temperature, acetic acid being removed under reduced pressure, being added water (20mL), 40%NaOH aqueous tune pH to 9, methylene chloride extracts (30mLx3), closes And organic phase, anhydrous sodium sulfate dry, filter, concentration, residue obtains white solid 1.7g, yield through silica gel post separation 53.4%.LC-MS (APCI): m/z=295.1 and 297.1 (M+1)⁺.¹H NMR(CDCl₃,300MHz)(δ/ppm):7.41(d, J=4.8Hz, 1H), 7.37 (s, 1H), 7.07 (d, J=6.6Hz, 1H), 6.72 (s, 1H), 2.14 (s, 3H), 1.90-1.85 (m,1H),0.90-0.78(m,4H).

The synthesis of step 7 compound 10.

Compound 9 (1.7g, 5.8mmol) is added into the 50mL three-necked flask equipped with magnetic agitation, vacuumizes simultaneously N₂It sets 3 times are changed, N₂It is instilled under atmosphere anhydrous THF (30mL), stirs dissolved clarification, be cooled to 0 DEG C, be slowly added dropwise into isopropylmagnesium chloride (4.3mL, 8.6mmol, 2M), insulated and stirred is reacted 1 hour, then by being full of CO₂The balloon of gas is slow into reaction solution It is passed through CO₂, react 1 hour, water (20mL) quenching reaction, 6M HCl/water liquid tune pH to 5, ethyl acetate extraction be added (30mLx3) merges organic phase, and anhydrous sodium sulfate dries, filters, and is concentrated, and residue Diethyl ether recrystallization obtains white solid 600mg, Yield 40.0%.LC-MS (APCI): m/z=261.1 (M+1)⁺.¹H NMR(DMSO-d₆,300MHz)(δ/ppm):9.17(s, 1H), 7.95 (d, J=4.8Hz, 1H), 7.70 (s, 1H), 7.52 (d, J=8.4Hz, 1H), 2.24 (s, 3H), 2.05-1.91 (m,1H),1.01-0.85(m,4H).

The synthesis of step 8 compound T-1.

Compound 10 (100mg, 0.384mmol) is added into the 50mL three-necked flask equipped with magnetic agitation, vacuumizes simultaneously N₂Displacement, N₂Dry methylene chloride (2mL) and DMF (2mL) are instilled under atmosphere by syringe, stirs dissolved clarification, reaction solution is cooled to It 0 DEG C, is slowly added dropwise into oxalyl chloride (dichloromethane solution of 0.33mL, 0.653mmol, 2M), drop finishes, and removes ice bath, and room temperature is stirred Mix reaction 1 hour.Evaporating solvent under reduced pressure is added dry methylene chloride (10mL), N₂Dissolved clarification is stirred under atmosphere, is cooled to 0 DEG C, slowly Be added dropwise to the dichloromethane solution (2mL) of compound 4 (94mg, 0.461mmol), be then added dropwise to DIPEA (0.19mL, 1.15mmol), ice bath is removed, reaction 2 hours is stirred at room temperature.Water (20mL) quenching reaction is added, separates organic layer, water phase dichloro Methane extracts (20mLx2), merges organic phase, and anhydrous sodium sulfate dries, filters, and is concentrated, and it is white that residue through silica gel post separation obtains class Color solid 60mg, yield 35.1%.LC-MS (APCI): m/z=446.4 (M+1)⁺.¹H NMR(CDCl₃,300MHz)(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=6.0Hz, 1H), 7.45 (s, 1H), 7.18 (d, J=9.0Hz, 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=5.1Hz, 6H), 0.90-0.81 (m, 4H)

2 5- of embodiment (4- cyclopropyl -1H- imidazole radicals -1- base)-N- (6- (4- isopropyl -4H-1,2,4- triazole -3- Base) pyridine -2- base) the fluoro- 4- methyl benzamide -6-d (compound T-2) of -2- preparation.

Specific synthesis step is as follows:

The synthesis of step 1 compound 12.

Compound 6 (2.0g, 9.8mmol) and heavy water (10mL) are added into the 20mL microwave tube equipped with magnetic agitation, stirs The heavy aqueous solution (0.817mL, 9.8mmol, 12M) for being slowly added to DCl is mixed down, finishes, is warming up to 160 under reaction mixture microwave DEG C, it reacts 1.5 hours.It is cooled to room temperature, is saturated NaHCO₃PH to 10 is adjusted, methylene chloride extracts (20mLx3), merge organic phase, Anhydrous sodium sulfate dries, filters, and is concentrated to give brown solid 1.59g, yield 79.1%.LC-MS (APCI): m/z=205.1 (M+ 1)⁺.¹H NMR(DMSO-d₆, 300MHz) and (s, the 3H) of (δ/ppm): 6.93 (d, J=7.2Hz, 1H), 4.94 (s, 2H), 1.99

The synthesis of step 2 compound 13.

Compound 12 (1.59g, 7.75mmol), toluene are successively added into the 100mL single-necked flask equipped with magnetic agitation (30mL) stirs dissolved clarification, is added dropwise to compound 5 (1.39g, 8.53mmol) and DIPEA (2.7mL, 16.28mmol), reaction solution It is heated to reflux 2 hours.It is cooled to room temperature, is added water (30mL), layering, organic phase is successively with saturation NH₄Cl aqueous (10mL) is satisfied And NaHCO₃Aqueous (10mL) and saturated salt solution (10mL) washing, anhydrous sodium sulfate dry, filter, and are concentrated, residue is just It is recrystallized in hexane, filters, dry to obtain off-white powder 1.5g, yield 67.4%.LC-MS (APCI): m/z=287.2& 289.2(M+1)⁺.

The synthesis of step 3 compound 14.

Compound 13 (1.5g, 5.22mmol) and dichloromethane are successively added into the 50mL single-necked flask equipped with magnetic agitation Alkane (10mL) stirs dissolved clarification and is cooled to 0 DEG C, is slowly added dropwise into formic acid (15mL) and acetic anhydride (2.1mL, 20.9mmol), instead Answer liquid N₂It is stirred to react at 0 DEG C 3 hours under atmosphere.Water (10mL) quenching reaction is added, 40%NaOH aqueous tune pH to 9 is separated Organic phase, water phase methylene chloride extract (20mLx2), merge organic phase, and anhydrous sodium sulfate dries, filters, and concentrate is directly used in It reacts in next step.LC-MS (APCI): m/z=315.1&317.1 (M+1)⁺.

The synthesis of step 4 compound 15.

Compound 14 (1.65g, 5.24mmol) and ice vinegar are successively added into the 50mL single-necked flask equipped with magnetic agitation Sour (15mL) stirs dissolved clarification, is added ammonium acetate (1.25g, 20.5mmol), N₂Reaction solution heated overnight at reflux under atmosphere.It is cooled to Room temperature removes acetic acid under reduced pressure, is added water (10mL), 40%NaOH aqueous tune pH to 9, and methylene chloride extracts (20mLx3), merges Organic phase, anhydrous sodium sulfate dry, filter, and residue obtains white solid 1.7g, yield 53.4% through silica gel post separation.LC-MS (APCI): m/z=296.1 and 298.1 (M+1)⁺.¹H NMR(CDCl₃,300MHz)(δ/ppm):7.38(s,1H),7.09(d,J =6.9Hz, 1H), 6.73 (s, 1H), 2.15 (s, 3H), 1.91-1.87 (m, 1H), 0.91-0.78 (m, 4H)

The synthesis of step 5 compound 16.

Compound 15 (820mg, 2.77mmol) is added into the 50mL three-necked flask equipped with magnetic agitation, vacuumizes simultaneously N₂ Displacement 3 times, N₂It is instilled under atmosphere anhydrous THF (15mL), stirs dissolved clarification, be cooled to 0 DEG C, be slowly added dropwise into isopropylmagnesium chloride (2.77mL, 5.54mmol, 2M), insulated and stirred is reacted 1 hour, then by being full of CO₂The balloon of gas delays into reaction solution Slowly it is passed through CO₂, react 1 hour, water (20mL) quenching reaction, 6M HCl/water liquid tune pH to 5, ethyl acetate extraction be added (30mLx3) merges organic phase, and anhydrous sodium sulfate dries, filters, and is concentrated, and residue obtains white solid with Diethyl ether recrystallization 200mg, yield 27.6%.LC-MS (APCI): m/z=262.1 (M+1)⁺.¹H NMR(DMSO-d₆,300MHz)(δ/ppm): 8.69 (s, 1H), 7.53 (s, 1H), 7.50 (d, J=8.7Hz, 1H), 2.24 (s, 3H), 1.96-1.93 (m, 1H), 0.96- 0.93(m,2H),0.80-0.77(m,2H).

The synthesis of step 6 compound T-2.

Compound 16 (130mg, 0.497mmol) is added into the 50mL three-necked flask equipped with magnetic agitation, vacuumizes simultaneously N₂It replaces three times, N₂Dry methylene chloride (2mL) and DMF (2mL) are respectively dropped by syringe under atmosphere, stir dissolved clarification, reaction Liquid is cooled to 0 DEG C, is slowly added dropwise oxalyl chloride (dichloromethane solution of 0.42mL, 0.846mmol, 2M), and drop finishes, and removes ice bath, Reaction 1 hour is stirred at room temperature.Evaporating solvent under reduced pressure is added dry methylene chloride (10mL), N₂Dissolved clarification is stirred under atmosphere, is cooled to 0 DEG C, the dichloromethane solution (2mL) into compound 3 (122mg, 0.597mmol) is slowly added dropwise, is then added dropwise to DIPEA (0.25mL, 1.49mmol) removes ice bath, reaction 2 hours is stirred at room temperature.Water (20mL) quenching reaction is added, separates organic layer, Water phase methylene chloride extracts (20mLx2), merges organic phase, and anhydrous sodium sulfate dries, filters, and is concentrated, and residue is through silicagel column point From off-white powder 60mg, yield 35.1%.LC-MS (APCI): m/z=447.4 (M+1)⁺.¹H NMR(CDCl₃, 300MHz) (δ/ppm): 9.06 (d, J=12.3Hz, 1H), 8.41-8.38 (m, 2H), 8.07 (d, J=5.7Hz, 1H), 7.93 (t, J=6.0Hz, 1H), 7.49 (s, 1H), 7.20 (d, J=9.6Hz, 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=5.1Hz, 6H), 0.91-0.81 (m, 4H)

3 5- of embodiment (4- cyclopropyl -1H- imidazole radicals -1- base)-N- (6- (4- (propyl- 2- base -1,1,1,3,3,3-d₆)- 4H-1,2,4- triazole -3- base) pyridine -2- base) the fluoro- 4- methyl benzamide (compound T-3) of -2- preparation.

Specific synthesis step is as follows:

The synthesis of step 1 compound 18.

Ammonium acetate (6.32g, 78mmol) and MeOD (50mL), N are added into the 50mL single-necked flask equipped with magnetic agitation₂ Temperature rising reflux 3 hours under atmosphere.It is concentrated to dryness, adds MeOD (10mL), be added with stirring acetone-d₆(1.0g, 15.6mmol) and NaBH₃CN (980mg, 15.6mmol), N₂Reaction is stirred at room temperature under atmosphere overnight.Water (10mL) is added to be quenched instead It answers, 6M hydrochloric acid tune pH to 2, ethyl acetate extracts (20mLx2), water phase 6M NaOH aqueous tune pH to 12, methylene chloride extraction (20mLx3) merges organic phase, and anhydrous sodium sulfate dries, filters, filtrate 2M hydrochloric acid methanol liquid tune pH to 2, is concentrated to dryness to change 18 hydrochloride of object is closed, is directly used in next step.

The synthesis of step 2 compound 19.

Compound 2 (300mg, 1.97mmol), toluene are successively added into the 50mL single-necked flask equipped with magnetic agitation (10mL), 18 hydrochloride of compound (642mg, 9.86mmol) and n,N-Dimethylformamide-dipropyl acetal (933mg, 5.32mmol), it is slowly added dropwise into acetic acid (296mg, 4.93mmol), finishes, N under stirring₂Reaction mixture is heated to reflux 3 under atmosphere Hour.It is cooled to room temperature, evaporating solvent under reduced pressure crosses silicagel column and obtains white solid 170mg, yield 41.2%.LC-MS (APCI): m/z=210.2 (M+1)⁺.¹H NMR(CDCl₃,300MHz)(δ/ppm):8.32(s,1H),7.62-7.56(m, 2H),6.58(dd,J₁=5.1Hz, J₂=1.2Hz, 1H), 5.59 (s, 1H), 4.51 (s, 2H)

The synthesis of step 3 compound T-3.

Compound 10 (130mg, 0.497mmol) is added into the 50mL three-necked flask equipped with magnetic agitation, vacuumizes simultaneously N₂Displacement, N₂Dry methylene chloride (2mL) and DMF (2mL) are respectively dropped by syringe under atmosphere, stir dissolved clarification, reacts liquid cooling But it to 0 DEG C, is slowly added dropwise into oxalyl chloride (dichloromethane solution of 0.42mL, 0.846mmol, 2M), drop finishes, and removes ice bath, room Temperature is stirred to react 1 hour.Evaporating solvent under reduced pressure is added dry methylene chloride (10mL), N₂Dissolved clarification is stirred under atmosphere, is cooled to 0 DEG C, The dichloromethane solution (2mL) into compound 19 (122mg, 0.597mmol) is slowly added dropwise, then be added dropwise DIPEA (0.25mL, 1.49mmol), ice bath is removed, reaction 2 hours is stirred at room temperature.Water (20mL) quenching reaction is added, separates organic layer, water phase dichloro Methane extracts (20mLx2), merges organic phase, and anhydrous sodium sulfate dries, filters, and is concentrated, and it is white that residue through silica gel post separation obtains class Color solid 60mg, yield 35.1%.LC-MS (APCI): m/z=452.4 (M+1)⁺.¹H NMR(CDCl₃,300MHz)(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=6.6Hz, 1H), 7.45 (s, 1H), 7.18 (d, J=9.0Hz, 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).

4 5- of embodiment (4- cyclopropyl -1H- imidazole radicals -1- base)-N- (6- (4- (propyl- 2- base-d₇) -4H-1,2,4- three Azoles -3- base) pyridine -2- base) the fluoro- 4- methyl benzamide (compound T-4) of -2- preparation.

Specific synthesis step is as follows:

The synthesis of step 1 compound 20.

Ammonium acetate (6.32g, 78mmol) and MeOD (50mL), N are added into the 50mL single-necked flask equipped with magnetic agitation₂ Temperature rising reflux 3 hours under atmosphere.It is concentrated to dryness, adds MeOD (10mL), be added with stirring acetone-d₆(1.0g, 15.6mmol) and NaBD₃CN (980mg, 15.6mmol), N₂Reaction is stirred at room temperature under atmosphere overnight.Water (10mL) is added to be quenched instead It answers, 6M hydrochloric acid tune pH to 2, ethyl acetate extracts (20mLx2), water phase 6M NaOH aqueous tune pH to 12, methylene chloride extraction (20mLx3) merges organic phase, and anhydrous sodium sulfate dries, filters, filtrate 2M hydrochloric acid methanol liquid tune pH to 2, is concentrated to dryness to change 20 hydrochloride of object is closed, is directly used in next step.

The synthesis of step 2 compound 21.

Compound 2 (300mg, 1.97mmol), toluene are successively added into the 50mL single-necked flask equipped with magnetic agitation (10mL), 20 hydrochloride of compound (642mg, 9.86mmol) and n,N-Dimethylformamide-dipropyl acetal (933mg, 5.32mmol), it is slowly added dropwise into acetic acid (296mg, 4.93mmol), finishes, N under stirring₂Reaction mixture is heated to reflux 3 under atmosphere Hour.It is cooled to room temperature, evaporating solvent under reduced pressure crosses silicagel column and obtains white solid 170mg, yield 41.2%.LC-MS (APCI): m/z=211.2 (M+1)⁺.

The synthesis of step 3 compound T-4.

Compound 10 (130mg, 0.497mmol) is added into the 50mL three-necked flask equipped with magnetic agitation, vacuumizes simultaneously N₂Displacement, N₂Dry methylene chloride (2mL) and DMF (2mL) are respectively dropped by syringe under atmosphere, stir dissolved clarification, reacts liquid cooling But it to 0 DEG C, is slowly added dropwise into oxalyl chloride (dichloromethane solution of 0.42mL, 0.846mmol, 2M), drop finishes, and removes ice bath, room Temperature is stirred to react 1 hour.Evaporating solvent under reduced pressure is added dry methylene chloride (10mL), N₂Dissolved clarification is stirred under atmosphere, is cooled to 0 DEG C, The dichloromethane solution (2mL) into compound 21 (122mg, 0.597mmol) is slowly added dropwise, then be added dropwise DIPEA (0.25mL, 1.49mmol), ice bath is removed, reaction 2 hours is stirred at room temperature.Water (20mL) quenching reaction is added, separates organic layer, water phase dichloro Methane extracts (20mLx2), merges organic phase, and anhydrous sodium sulfate dries, filters, and is concentrated, residue is through the isolated class of silicagel column White solid 60mg, yield 35.1%.LC-MS (APCI): m/z=453.4 (M+1)⁺.¹H NMR(CDCl₃,300MHz)(δ/ Ppm): 9.06 (d, J=12.0Hz, 1H), 8.41 (d, J=6.9Hz, 1H), 8.38 (s, 1H), 8.09 (d, J=5.7Hz, 2H), 7.94 (t, J=6.0Hz, 1H), 7.47 (s, 1H), 7.21 (d, J=9.3Hz, 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 inhibitory activity

Reagent and consumptive material:

ASK1 (Invitrogen catalog number (Cat.No.) PV3809), ATP (Sigma, catalog number (Cat.No.) A7699), DMSO (Sigma, catalog number (Cat.No.) D8418-1L), 384 orifice plates (Greiner, catalog number (Cat.No.) 784076), HTRF KinEASE-STK Kit kit (Cisbio, mesh Record PV3809), 5x kinase buffer liquid A (Life Technologies, catalog number (Cat.No.) PV3186), 199 (Life of kinases tracer Technologies, catalog number (Cat.No.) PV5830),Eu-anti-GST antibody (Life Technologies, catalogue Number PV5594).

Specific experiment method:

Compound is prepared: test-compound being dissolved in DMSO and is made into 20mM mother liquor.Then, in DMSO 3 times of constant gradient it is dilute It releases, dilutes ten times.10 times are diluted with buffer again when dosing.

ASK1 kinase assay: in 5x kinase buffer liquid A, the change for the various concentration that ASK1 kinases and beforehand dilution are prepared It closes object to mix 10 minutes, each concentration duplicate hole.Corresponding substrate and ATP is added, room temperature reaction (is provided with yin and yang attribute in 20 minutes Control: feminine gender is blank control, and the positive is Tarceva).Detection reagent (HTRF KinEASE-STK Kit is added in end of reaction Reagent in kit), after incubation at room temperature 30 minutes, is detected by Evnvision microplate reader, measure the present invention in each concentration Enzyme activity in the presence of compound, and the inhibitory activity of the compounds on enzyme activities of various concentration is calculated, later according to four parameters Equation is fitted according to inhibitory activity of 5.0 software of Graphpad to enzyme activity under various concentration compound, calculates IC₅₀ Value.

The compounds of this invention is tested in above-mentioned kinase inhibition assay and without deuterated compound Selonsertib, It was found that the compounds of this invention has more potent or comparable activity to ASK1 kinases.Suppression of the representative embodiment compound to kinases The result of production is summarized in as in the following table 1.

Table 1

Embodiment compound	ASK1 IC₅₀(nM)
		Selonsertib	1.10
T-1	1.00
		T-2	1.12
T-3	1.12
		T-4	1.03

(2) metabolic stability is evaluated

Microsomal assay: people's hepatomicrosome: 0.5mg/mL, Xenotech；Rat liver microsomes: 0.5mg/mL, Xenotech；Mouse Liver Microsomes: 0.5mg/mL, Xenotech；Coenzyme (NADPH/NADH): 1mM, Sigma Life Science；Magnesium chloride: 5mM, 100mM phosphate buffer (pH 7.4).

The preparation of stock solution: precision weighs the powder of a certain amount of embodiment compound, and is dissolved to respectively with DMSO 5mM。

The preparation of phosphate buffer (100mM, pH7.4): take the 150mL prepared in advance 0.5M potassium dihydrogen phosphate and The 0.5M dipotassium hydrogen phosphate solution of 700mL mixes, then adjusts mixed liquor pH value to 7.4 with 0.5M dipotassium hydrogen phosphate solution, uses It is preceding to dilute 5 times with ultrapure water, magnesium chloride is added, phosphate buffer (100mM) is obtained, wherein potassium phosphate containing 100mM, 3.3mM Magnesium chloride, pH 7.4.

It prepares NADPH regenerative system solution and (contains 6.5mM NADP, 16.5mM G-6-P, 3U/mL G-6-P D, 3.3mM Magnesium chloride), using it is preposition in it is wet on ice.

Prepare terminate liquid: the acetonitrile containing 50ng/mL Propranolol Hydrochloride and 200ng/mL orinase (internal standard) is molten Liquid.It takes 25057.5 μ L phosphate buffers (pH7.4) into 50mL centrifuge tube, is separately added into 812.5 μ L people, rat and mouse Hepatomicrosome mixes, and obtains the hepatomicrosome dilution that protein concentration is 0.625mg/mL.The incubation of sample: with containing 70% second The stock solution of respective compound is diluted to 0.25mM by the aqueous solution of nitrile respectively, spare as working solution.Take 398 μ L's respectively People's hepatomicrosome, rat liver microsomes or Mouse Liver Microsomes dilution are added 96 holes and are incubated in plate (N=2), are separately added into 2 In the working solution of μ L 0.25mM, mix.

The measurement of metabolic stability: the terminate liquid of 300 μ L pre-cooling is added in every hole of 96 hole deep-well plates, is placed in ice On, as termination plate.96 holes are incubated for plate and NADPH regenerative system is placed in 37 DEG C of water baths, 100 revs/min of concussions are incubated in advance 5min.80 μ L Incubating Solutions addition termination plate is taken out from the every hole of plate is incubated for, mixes, supplements 20 μ L NADPH regenerative system solution, make For 0min sample.Again to the NADPH regenerative system solution for being incubated for 80 μ L of the every hole addition of plate, starting reaction starts timing.Correspondingization The reaction density for closing object is 1 μM, protein concentration 0.5mg/mL.When reacting 10,30,90min, 100 μ L is respectively taken to react Liquid is added in termination plate, and vortex 3min terminates reaction.Termination plate is centrifuged 10min under the conditions of 5000 × g, 4 DEG C.Take 100 μ L Supernatant is mixed to being previously added in 96 orifice plates of 100 μ L distilled water, carries out sample analysis using LC-MS/MS.

Data analysis: by LC-MS/MS system detection respective compound and interior target peak area, calculate compound with it is interior Mark peak area ratio.Slope is measured by the natural logrithm of the percentage of compound surplus and time mapping, and according to following Formula calculates t_1/2And CL_int, wherein V/M is equal to 1/ protein concentration.

The compounds of this invention and testting simultaneously without deuterated compound Selonsertib is compared, evaluates it in people, big The metabolic stability of mouse and Mouse Liver Microsomes.The half-life period of index as metabolic stability and liver clearance rate such as table 2 It is shown.As shown in table 2, people, rat and Mouse Liver Microsomes experiment in, by with without deuterated compound Selonsertib control, the compounds of this invention can be obviously improved metabolic stability.

Table 2

(3) pharmacokinetics in rats is tested

6 male Sprague-Dawley rats, 7-8 week old, weight about 210g are divided into 2 groups, every group 3, through vein or The compound (oral 10mg/kg) of oral single dosage, compares its pharmacokinetic difference.

Rat is raised using standard feed, gives water.Test is fasted for first 16 hours.Drug is sub- with PEG400 and diformazan Sulfone dissolution.Eye socket blood sampling, the time point of blood sampling are 0.25 hour, 0.5 hour, 1 hour, 2 hours, 4 0.083 hour after administration Hour, 6 hours, 8 hours, 12 hours and 24 hours.

Rat sucks of short duration anesthesia after ether, and eye socket acquires 300 μ L sample of blood in test tube.There is 30 μ L, 1% heparinate in test tube Solution.Before use, test tube is stayed overnight in 60 DEG C of drying.After the last one time point blood specimen collection completion, rat etherization After put to death.

It after blood specimen collection, leniently overturns test tube at least 5 times, is placed on ice after guaranteeing mixing sufficiently immediately.Blood sample is 4 DEG C 5000rpm is centrifuged 5 minutes, and blood plasma is separated with red blood cell.100 μ L blood plasma are sucked out to clean plastic centrifuge with pipettor Guan Zhong indicates title and the time point of compound.Blood plasma is stored in -80 DEG C before being analyzed.Blood plasma is measured with LC-MS/MS The concentration of middle the compounds of this invention.Pharmacokinetic parameter is based on every animal blood concentration in different time points into calculating.

Experiment, which shows the compounds of this invention in animal body, has better pharmacokinetic property, therefore has more preferable Pharmacodynamics and control distant effect.

The above content is a further detailed description of the present invention in conjunction with specific preferred embodiments, and it cannot be said that Specific implementation of the invention is only limited to these instructions.For those of ordinary skill in the art to which the present invention belongs, exist Under the premise of not departing from present inventive concept, a number of simple deductions or replacements can also be made, all shall be regarded as belonging to of the invention Protection scope.

Claims

1. 1,2, the 4- triazole compounds or its pharmaceutically acceptable salt of a kind of formula (I):

Wherein,

X¹、X²、X³It is CH each independently₃、CH₂D、CHD₂Or CD₃；

Condition is if X¹、X²And X³Each is CH₃, then R¹、R²、R³、R⁴、R⁵、R⁶、R⁷、R⁸、Y¹、Y²、Y³、Y⁴、Y⁵And Y⁶In At least one be deuterium.

2. according to claim 11,2,4- triazole compounds or its pharmaceutically acceptable salt, wherein R¹、R³ And R⁴It is deuterium.

3. according to claim 1 or 21,2,4- triazole compounds or its pharmaceutically acceptable salt, wherein R⁵ It is deuterium.

4. according to any one of claim 1-3 1,2,4- triazole compounds or its pharmaceutically acceptable salt, Wherein, X¹And X²It is CD₃。

5. 1,2,4- triazole compounds or its pharmaceutically acceptable salt described in any one of -4 according to claim 1, Wherein, Y¹It is deuterium.

6. according to claim 11,2,4- triazole compounds or its pharmaceutically acceptable salt, wherein describedization Object is closed to be selected from:

7. a kind of pharmaceutical composition, containing pharmaceutically acceptable excipient and as claimed in any one of claims 1 to 61, 2,4- triazole compound or its pharmaceutically acceptable salt.

8. a kind of preparation method of pharmaceutical composition as claimed in claim 7, comprising: by pharmaceutically acceptable excipient with As claimed in any one of claims 1 to 61,2,4- triazole compounds or its pharmaceutically acceptable salt are mixed, from And form pharmaceutical composition.

9. described in any one of claims 1-6 1 is prepared, 2,4- triazole compounds or its pharmaceutically acceptable salt, or The method that pharmaceutical composition described in claim 7 or 8 is used to treat the drug at least partly by the ASK1 disease mediated.

10. method as claimed in claim 9 further includes that another therapeutic agent is administered；Preferably, wherein another therapeutic agent is LOX2 inhibitor.

11. method described in claim 9 or 10, wherein the disease be diabetes, diabetic nephropathy, nephrosis, kidney fibrosis, Pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), liver fibrosis, pulmonary hypertension, nonalcoholic fatty liver hepatitis, hepatopathy, alcohol Property hepatopathy, inflammatory conditions, autoimmune disease, proliferative diseases, graft rejection, be related to the impaired disease of cartilage metabolism, elder generation Nature cartilage deformity or disease relevant to IL6 hypersecretion.

12. method described in claim 11, wherein the disease is nonalcoholic steatohepatitis or alcoholic liver disease.

13. method described in claim 11, wherein the disease is pulmonary hypertension or pulmonary fibrosis.

14. method described in claim 11, wherein the disease is diabetic nephropathy, nephrosis or kidney fibrosis.