CN109232538A - A kind of 1,2,4- triazole compound - Google Patents
A kind of 1,2,4- triazole compound Download PDFInfo
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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
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;
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, 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+(C1-4Alkyl)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 t1/2And CLint, 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 (14)
1. 1,2, the 4- triazole compounds or its pharmaceutically acceptable salt of a kind of formula (I):
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;
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.
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CN102482257A (en) * | 2009-07-13 | 2012-05-30 | 吉利德科学股份有限公司 | Apoptosis signal-regulating kinase inhibitors |
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CN104080771A (en) * | 2012-01-27 | 2014-10-01 | 吉利德科学公司 | Apoptosis signal-regulating kinase inhibitor |
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