A compound as a thyroid hormone beta receptor agonist and its use

Abstract

The present invention relates to a compound that acts as a thyroid hormone beta receptor agonist and its use, and further relates to a compound represented by formula (1) or a pharmaceutically acceptable form thereof, a pharmaceutical composition comprising the same, and a method for preparing the same. The compound or pharmaceutical composition described herein can be used to prepare a medicament for preventing, treating, or alleviating diseases regulated by thyroid hormone beta receptors. #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry and relates to compounds that serve as thyroid hormone beta receptor agonists, pharmaceutical compositions containing the compounds, methods for preparing the compounds, and uses of the compounds in preparing drugs for preventing, treating or alleviating diseases regulated by thyroid hormone beta receptors. Background Art

[0002] Thyroid hormone (TH) is synthesized in the thyroid gland in response to thyroid-stimulating hormone (TSH) secreted by the pituitary gland. Thyroid hormone plays a crucial role in regulating growth, development, metabolism, and matrix homeostasis. Thyroid hormone exerts its function by binding to thyroid hormone receptors (THRs). THRs belong to the nuclear receptor superfamily. Nuclear receptors form heterodimers with their common ligand, the retinoid X receptor, and function as ligand-induced transcription factors. Like other nuclear receptors, THRs possess ligand-binding and DNA-binding domains and regulate gene expression through ligand-dependent interactions with DNA-responsive elements (thyroid response elements, THREs).

[0003] There are currently two THR isoforms: THRα and THRβ. THRα is primarily distributed in cardiac tissue and plays an important regulatory role in cardiac function. THRβ is primarily expressed in the liver and pituitary gland, regulating the metabolism of fatty acids and cholesterol, as well as the secretion of thyroid-stimulating hormone. Both THRα and THRβ are expressed in brown adipose tissue (BAT), playing an important role in regulating basal oxygen consumption, fat storage, lipogenesis, and lipolysis (Oppenheimer et al., J. Clin. Invest. 87(1):125-32 (1991)).

[0004] THR agonists increase metabolic rate, oxygen consumption, and heat production, promote cholesterol metabolism into bile acids, and reduce lipoprotein levels associated with atherosclerosis. The liver and heart are the primary target organs of THR agonists. In the liver, they primarily regulate genes involved in the synthesis and metabolism of fatty acids and cholesterol, and affect carbohydrates by increasing glycogenolysis and gluconeogenesis and reducing the effects of insulin. In the heart, they can reduce systemic vascular resistance, increase blood volume, and produce inotropic and chronotropic effects.

[0005] THRβ agonists can also enhance cellular lipid metabolism and lower cholesterol and blood lipids. Therefore, research and development of THRβ agonists for the treatment and / or prevention of diseases regulated by thyroid hormone receptors is of great significance. Summary of the Invention

[0006] Through extensive research, the present invention has discovered a series of compounds that serve as thyroid hormone beta receptor agonists and have potential value in preventing and / or treating diseases regulated by thyroid hormone beta receptors.

[0007] In a first aspect, the present invention provides a compound having the structure of formula (1) or a pharmaceutically acceptable form thereof:

[0008]

[0009] in,

[0010] Ring A is selected from C 6-10 Aryl, 5-12 membered heteroaryl, C 5-10 Cycloalkyl or 5-10 membered heterocyclic group, wherein the ring A is optionally substituted by one or more R4, wherein the R4 is independently selected from halogen, cyano, amino, nitro, hydroxyl, -OCF3, -NH(C 1-4 Alkyl), -N(C 1-4 alkyl)2、-C(=O)-NH2、-C(=O)NH(C 1-4 alkyl), -C(=O)N(C 1-4 alkyl)2, -C(=O)-C 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy or C 3-6 Cycloalkyl,

[0011] When two or more R4 are present, two R4 are optionally taken together with the atoms to which they are attached to form C 6-10 Aryl, 5-12 membered heteroaryl, C 5-10 Cycloalkyl or 5-10 membered heterocyclic group,

[0012] R1 is selected from hydrogen, C 1-6 Alkyl or C 3-6 Cycloalkyl, the alkyl or cycloalkyl being optionally substituted by one or more independently selected from deuterium, halogen, cyano, amino, nitro, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or C 6-10 The substituents of the aryl group are substituted,

[0013] R2 and R3 are independently selected from hydrogen, halogen, cyano, amino, nitro, hydroxy, C 1-6 Alkyl or C 1-6 Alkoxy,

[0014] X is selected from -C 1-6 Alkylene-, -C(=O)- or -C(=O)-C 1-6Alkylene-, the alkylene is optionally substituted by one or more independently selected from halogen, cyano, amino, nitro, hydroxyl, C 1-6 Alkyl or C 1-6 Alkoxy substituents are substituted,

[0015] The pharmaceutically acceptable form is selected from pharmaceutically acceptable salts, esters, stereoisomers, tautomers, solvates, N-oxides, isotopically labeled forms, metabolites, and prodrugs.

[0016] In some embodiments, the ring A in the compound of formula (1) or a pharmaceutically acceptable form thereof is selected from C 6-10 Aryl, 5-12 membered heteroaryl or C 5-10 Cycloalkyl, wherein the ring A is optionally substituted by one or more R4, wherein the R4 is independently selected from halogen, cyano, amino, nitro, hydroxyl, -OCF3, -NH(C 1-4 alkyl), -C(=O)-NH2, -C(=O)NH(C 1-4 alkyl), -C(=O)-C 1-4 Alkyl, C 1-6 Alkyl or C 1-6 Alkoxy.

[0017] In some preferred embodiments, the ring A in the compound of formula (1) or its pharmaceutically acceptable form is selected from C 6-10 Aryl, 5-10 membered heteroaryl or C 5-8 Cycloalkyl, wherein the ring A is optionally substituted by one or more R4, wherein the R4 is independently selected from halogen, cyano, amino, nitro, hydroxyl, -OCF3 or C 1-4 alkyl.

[0018] In some more preferred embodiments, ring A in the compound of formula (1) or a pharmaceutically acceptable form thereof is selected from phenyl, pyridyl, cyclopentyl, cyclohexyl or bicyclo[2.2.1]heptyl, and the ring A is optionally substituted by one or more R4, and the R4 is independently selected from fluorine, chlorine, bromine, cyano, amino, nitro, hydroxyl or methyl.

[0019] In some particularly preferred embodiments, ring A in the compound of formula (1) or a pharmaceutically acceptable form thereof is selected from

[0020] In some embodiments, R1 in the compound of formula (1) or a pharmaceutically acceptable form thereof is selected from hydrogen or C 1-6 Alkyl, said alkyl being optionally substituted by one or more independently selected from deuterium, halogen, cyano, amino, nitro, hydroxyl, C 1-6 Alkyl or C 6-10 The substituents of the aryl group are substituted.

[0021] In some preferred embodiments, R1 in the compound of formula (1) or its pharmaceutically acceptable form is selected from hydrogen or C 1-4 Alkyl, optionally substituted with one or more substituents independently selected from deuterium, fluorine, chlorine, cyano, amino, nitro, hydroxyl, or phenyl.

[0022] In some more preferred embodiments, R1 in the compound of formula (1) or a pharmaceutically acceptable form thereof is selected from hydrogen, methyl, -CD3, ethyl,

[0023] In some embodiments, R2 and R3 in the compound of formula (1) or a pharmaceutically acceptable form thereof are independently selected from hydrogen, halogen, cyano, amino, nitro, hydroxyl or C 1-6 alkyl.

[0024] In some preferred embodiments, R2 and R3 in the compound of formula (1) or its pharmaceutically acceptable form are independently selected from halogen or C 1-4 alkyl.

[0025] In some more preferred embodiments, R2 and R3 in the compound of formula (1) above or a pharmaceutically acceptable form thereof are independently selected from Cl, Br or methyl.

[0026] In some embodiments, X in the compound of formula (1) or a pharmaceutically acceptable form thereof is selected from -C 1-4 Alkylene-, -C(=O)- or -C(=O)-C 1-4 Alkylene-, the alkylene is optionally substituted by one or more independently selected from halogen, cyano, amino, nitro, hydroxyl or C 1-4 The alkyl group is substituted with a substituent.

[0027] In some preferred embodiments, X in the compound of formula (1) or a pharmaceutically acceptable form thereof is selected from -CH2-, -CH2CH2-, -CH2CH2CH2-, -C(=O)-, -C(=O)-CH2- or -CH(CH3)-.

[0028] In some embodiments, the compound of formula (1) or a pharmaceutically acceptable form thereof is a compound having a structure of formula (2) or formula (3) or a pharmaceutically acceptable form thereof:

[0029]

[0030] wherein R1, R2, R3, R4 and X are as defined in formula (1), X1 is selected from N or CR4, m is selected from 1, 2 or 3, and n is selected from 0, 1, 2 or 3.

[0031] Those skilled in the art will appreciate that the present invention encompasses compounds obtained by any combination of the various embodiments. Embodiments obtained by combining the technical features or preferred technical features in one embodiment with the technical features or preferred technical features in another embodiment are also included within the scope of the present invention.

[0032] In a second aspect, the present invention further provides a compound or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, wherein the compound is selected from:

[0033]

[0034] In a third aspect, the present invention provides a method for preparing a compound having the structure of formula (1), comprising the following steps:

[0035] Step 1: Synthesis of intermediates M1 and M2

[0036]

[0037] (a) using the compound of formula I as a starting material to react with hydrazine hydrate to obtain a compound of formula II;

[0038] (b) The compound represented by the general formula II is reacted with phosphorus oxychloride to obtain the compound represented by the general formula M1.

[0039]

[0040] (c) introducing a protecting group into the amino group of the compound represented by the general formula III to obtain a compound represented by the general formula M2;

[0041] wherein R2, R3 and ring A are as defined in formula (1); and PG is an amino protecting group selected from tert-butyloxycarbonyl, benzyloxycarbonyl, phthalimide, p-toluenesulfonyl, methylsulfonyl, mesityl, formyl or trifluoroacetyl, preferably tert-butyloxycarbonyl.

[0042] Step 2-1: Synthesis route 1 of the compound represented by formula (1)

[0043]

[0044] (d) reacting the compound represented by the general formula M1 with the compound represented by the general formula III in the presence of a copper halide and a base to obtain the compound represented by the general formula IV;

[0045] (e) reacting the compound represented by the general formula IV with the compound represented by the general formula V in the presence of a base to obtain the compound represented by the general formula VI;

[0046] (f) reacting the compound represented by the general formula VI in the presence of a strong base to obtain the compound represented by the general formula VII;

[0047] (g) reacting the compound represented by the general formula VII with hydrazine hydrate to obtain the compound represented by the general formula VIII;

[0048] (h) reacting the compound represented by the general formula VIII with phosgene, triphosgene, carbonyldiimidazole or N,N'-disuccinimidyl carbonate in the presence of a base to obtain the compound represented by the formula (1);

[0049] wherein R1 is hydrogen, R2, R3, X and ring A are as defined in formula (1), and Y is selected from F, Cl, Br, I, OTs or OMs, preferably Br.

[0050] In some embodiments, (d) in step 2-1 above is carried out in the presence of a copper halide and a base, wherein the copper halide is selected from cuprous chloride, cuprous bromide, cuprous iodide, cupric chloride, cupric bromide or cupric iodide, preferably cuprous iodide; and the base is selected from triethylamine, diisopropylethylamine, pyridine, imidazole, 1,8-diazabicycloundec-7-ene, sodium carbonate, potassium carbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium ethoxide, preferably potassium carbonate.

[0051] In some embodiments, (e) in the above step 2-1 is carried out in the presence of a base, and the base is selected from triethylamine, diisopropylethylamine, pyridine, imidazole, 1,8-diazabicycloundec-7-ene, sodium carbonate, potassium carbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium ethoxide, preferably potassium carbonate.

[0052] In some embodiments, (f) in the above step 2-1 is carried out in the presence of a base, and the base is selected from triethylamine, diisopropylethylamine, pyridine, imidazole, 1,8-diazabicycloundec-7-ene, sodium carbonate, potassium carbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium ethoxide, sodium acetate, preferably sodium acetate.

[0053] In some embodiments, (h) in the above step 2-1 is carried out in the presence of a base, and the base is selected from triethylamine, diisopropylethylamine, pyridine, imidazole, 1,8-diazabicycloundec-7-ene, sodium carbonate, potassium carbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium ethoxide, preferably triethylamine.

[0054] Step 2-2: Synthesis route 2 of the compound represented by formula (1)

[0055]

[0056] (d) reacting the compound represented by the general formula M1 with the compound represented by the general formula M2 in the presence of a copper halide and a base to obtain a compound represented by the general formula VIV;

[0057] (e) reacting the compound represented by the general formula VIV with the compound represented by the general formula V in the presence of a base to obtain the compound represented by the general formula X;

[0058] (f) reacting the compound represented by the general formula X in the presence of a base to obtain a compound represented by the general formula XI;

[0059] (i) reacting the compound represented by the general formula XI with a halogenated hydrocarbon in the presence of a base to obtain a compound represented by the general formula XII;

[0060] (j) removing the protecting group on the amino group of the compound represented by the general formula XII to obtain the compound represented by the general formula XIII;

[0061] (g) reacting the compound represented by the general formula XIII with hydrazine hydrate to obtain the compound represented by the general formula XIV;

[0062] (h) reacting the compound represented by the general formula XIV with phosgene, triphosgene, carbonyldiimidazole or N,N'-disuccinimidyl carbonate in the presence of a base to obtain the compound represented by the formula (1);

[0063] Wherein, R1 is selected from C 1-6 Alkyl or C 3-6 Cycloalkyl, the alkyl or cycloalkyl being optionally substituted by one or more independently selected from deuterium, halogen, cyano, amino, nitro, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or C 6-10 The substituents of the aryl group are substituted, R2, R3, X and ring A are as defined in formula (1), and PG and Y are as defined above.

[0064] In some embodiments, (d) in step 2-2 above is carried out in the presence of a copper halide and a base, wherein the copper halide is selected from cuprous chloride, cuprous bromide, cuprous iodide, cupric chloride, cupric bromide or cupric iodide, preferably cuprous iodide; and the base is selected from triethylamine, diisopropylethylamine, pyridine, imidazole, 1,8-diazabicycloundec-7-ene, sodium carbonate, potassium carbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium ethoxide, preferably potassium carbonate.

[0065] In some embodiments, (e) in the above step 2-2 is carried out in the presence of a base, and the base is selected from triethylamine, diisopropylethylamine, pyridine, imidazole, 1,8-diazabicycloundec-7-ene, sodium carbonate, potassium carbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium ethoxide, preferably potassium carbonate.

[0066] In some embodiments, (f) in the above step 2-2 is carried out in the presence of a base, and the base is selected from triethylamine, diisopropylethylamine, pyridine, imidazole, 1,8-diazabicycloundec-7-ene, sodium carbonate, potassium carbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium ethoxide, sodium acetate, preferably sodium acetate.

[0067] In some embodiments, (i) in step 2-2 above is carried out in the presence of a halogenated hydrocarbon and a base, wherein the halogenated hydrocarbon is selected from a chlorohydrocarbon, a bromohydrocarbon or an iodohydrocarbon, preferably an iodohydrocarbon; and the base is selected from triethylamine, diisopropylethylamine, pyridine, imidazole, 1,8-diazabicycloundec-7-ene, sodium carbonate, potassium carbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium ethoxide, preferably potassium carbonate.

[0068] In some embodiments, (h) in step 2-2 above is carried out in the presence of a base, and the base is selected from triethylamine, diisopropylethylamine, pyridine, imidazole, 1,8-diazabicycloundec-7-ene, sodium carbonate, potassium carbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium ethoxide, preferably triethylamine.

[0069] In a fourth aspect, the present invention provides a pharmaceutical composition comprising at least one compound of formula (1) to formula (3) above or a pharmaceutically acceptable form thereof, and one or more pharmaceutically acceptable carriers.

[0070] In a fifth aspect, the present invention provides the compounds of formula (1) to formula (3) above or their pharmaceutically acceptable forms, or the pharmaceutical compositions above, which are used as thyroid hormone β receptor agonists for preventing and / or treating diseases or conditions mediated at least in part by thyroid hormone β receptors.

[0071] In a sixth aspect, the present invention provides the use of the compounds of formula (1) to formula (3) above or their pharmaceutically acceptable forms or the pharmaceutical compositions above in the preparation of a medicament for preventing and / or treating a disease or condition mediated at least in part by thyroid hormone beta receptor (e.g., metabolic diseases, such as non-alcoholic fatty liver disease, dyslipidemia, atherosclerosis or hypothyroidism).

[0072] In a seventh aspect, the present invention provides a method for preventing and / or treating a disease or condition mediated at least in part by a thyroid hormone beta receptor, comprising the following steps: administering a preventively and / or therapeutically effective amount of the above-mentioned compounds of formula (1) to formula (3) or a pharmaceutically acceptable form thereof or the above-mentioned pharmaceutical composition to an individual in need thereof.

[0073] The present invention is not to be limited to the particular embodiments described herein; it is also to be understood that the terminology used herein is for the purpose of describing and not limiting particular embodiments.

[0074] Definition of terms

[0075] Unless otherwise specified, the following terms have the following meanings in the present invention.

[0076] The terms "comprises," "including," "having," or "containing" or any other variations thereof are intended to cover a non-exclusive or open-ended inclusion. For example, a composition, method, or apparatus that comprises a list of elements is not necessarily limited to only the elements expressly listed but may include other elements not expressly listed or inherent to such composition, method, or apparatus.

[0077] When the lower and upper limits of a numerical range are disclosed, any value or sub-range falling within the range is specifically disclosed. In particular, each numerical range of a parameter disclosed herein (e.g., in the form of "about a to b," or equivalently "approximately a to b," or equivalently "about a b") should be understood to encompass every value and sub-range therein. For example, "C 1-4 " should be understood to include any sub-ranges and every point value therein, such as C 2-4 、C 3-4 、C 1-2 、C 1-3 、C 1-4 etc., as well as C1, C2, C3, C4, etc. For another example, "5-10 yuan" should be understood to cover any sub-range and every point value therein, such as 5-6 yuan, 5-7 yuan, 5-8 yuan, 5-9 yuan, 6-7 yuan, 6-8 yuan, etc., as well as 5, 6, 7, 8, 9, 10 yuan, etc.

[0078] The term "substituted" and its other variant forms in this article refer to that one or more (such as 1, 2, 3 or 4) atoms or atomic groups (such as hydrogen atoms) on the specified atom are replaced by other equivalents, provided that the normal valence of the specified atom or atomic group in the current situation is not exceeded and a stable compound can be formed. If an atom or atomic group is described as "optionally substituted by...", it can be substituted or unsubstituted. Unless otherwise indicated, the attachment site of a substituent herein can be from any suitable position of the substituent. When the connecting bond in a substituent is shown as a chemical bond between two atoms connected to each other in a ring system, it means that the substituent can be connected to any ring-forming atom in the ring system.

[0079] The term "pharmaceutical composition" refers to a composition that can be used as a medicine, which contains a pharmaceutically active ingredient (or therapeutic agent) and, optionally, one or more pharmaceutically acceptable carriers. The term "pharmaceutically acceptable carrier" refers to an excipient that is administered together with a therapeutic agent and is suitable, within the scope of sound medical judgment, for contact with the tissues of humans and / or other animals without excessive toxicity, irritation, allergic reaction, or other problems or complications commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable carriers that can be used in the present invention include, but are not limited to: a) diluents; b) lubricants; c) binders; d) disintegrants; e) absorbents, colorants, flavorings, and / or sweeteners; f) emulsifiers or dispersants; and / or g) substances that enhance the absorption of the compound, etc.

[0080] Above-mentioned pharmaceutical composition can act systemically and / or act topically.For this purpose, they can be administered by suitable approach, for example by parenteral, local, intravenous, oral, subcutaneous, intraarterial, intradermal, percutaneous, rectal, intracranial, intraperitoneal, intranasal, intramuscular approach or as inhalant administration.

[0081] The above-mentioned route of administration can be achieved through suitable dosage forms. The dosage forms that can be used in the present invention include, but are not limited to, tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, syrups, etc.

[0082] When administered orally, the pharmaceutical composition can be prepared into any orally acceptable preparation form, including but not limited to tablets, capsules, aqueous solutions, aqueous suspensions, and the like.

[0083] The pharmaceutical composition can also be administered in the form of a sterile injection, including a sterile water or oil suspension for injection, or a sterile water or oil solution for injection. The carriers that can be used include, but are not limited to, water, Ringer's solution, and isotonic sodium chloride solution. Additionally, sterile, fixed oils, such as monoglycerides or diglycerides, can also be used as solvents or suspending media.

[0084] The pharmaceutical composition may contain 0.01 mg to 1000 mg of at least one compound of formula (1) to formula (3) or a pharmaceutically acceptable form thereof.

[0085] The term "disease or condition mediated at least in part by the thyroid hormone beta receptor" refers to a disease whose pathogenesis involves at least in part a factor related to the thyroid hormone beta receptor, such as a metabolic disease, such as non-alcoholic fatty liver disease, dyslipidemia, atherosclerosis, or hypothyroidism.

[0086] The term "effective amount" refers to a dose that can induce a biological or medical response in cells, tissues, organs or organisms (eg, individuals) and is sufficient to achieve the desired preventive and / or therapeutic effect.

[0087] The dosage regimen can be adjusted to provide the optimal desired response. For example, the drug may be administered in a single dose, divided doses may be administered over time, or the dosage may be proportionally reduced or increased based on the actual situation. It will be understood that for any particular individual, the specific dosage regimen should be adjusted according to the needs and the professional judgment of the person administering the composition or supervising the administration of the composition.

[0088] The term "in need of" refers to the physician's or other health care provider's judgment that an individual needs or will benefit from a preventive and / or therapeutic procedure, which judgment is based on various factors within the physician's or other health care provider's area of expertise.

[0089] The term "individual" (or subject) refers to a human or non-human animal. The individual of the present invention includes individuals (patients) suffering from a disease and / or condition and normal individuals. The non-human animals of the present invention include all vertebrates, such as non-mammals, such as birds, amphibians, reptiles, etc., and mammals, such as non-human primates, livestock and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).

[0090] The term "treat" refers to the alleviation or elimination of the disease or condition being treated. If a subject receives a therapeutic amount of a compound of the present invention or a pharmaceutically acceptable form thereof or a pharmaceutical composition of the present invention, and at least one indicator and symptom of the subject shows observable and / or detectable relief and / or improvement, the subject is said to have been successfully "treated". It is understood that treatment includes not only complete treatment, but also includes not achieving complete treatment but achieving some biologically or medically relevant results. Specifically, "treat" means that the compound of the present invention or a pharmaceutically acceptable form thereof or a pharmaceutical composition of the present invention can achieve at least one of the following effects, for example: (1) preventing the occurrence of the disease in an animal that may be predisposed to the disease but has not yet experienced or displayed the pathology or symptomology of the disease; (2) inhibiting the disease in an animal that is experiencing or displaying the pathology or symptomology of the disease (i.e., preventing the further development of the pathology and / or symptomology); (3) ameliorating the disease in an animal that is experiencing or displaying the pathology or symptomology of the disease (i.e., reversing the pathology and / or symptomology).

[0091] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention that is substantially non-toxic to living organisms. Pharmaceutically acceptable salts generally include, but are not limited to, salts formed by reacting a compound of the present invention with a pharmaceutically acceptable inorganic / organic acid or inorganic / organic base, such salts also known as acid addition salts or base addition salts. For a review of suitable salts, see, for example, Jusiak, Soczewinski, et al., Remington's Pharmaceutical Sciences [M], Mack Publishing Company, 2005 and Stahl, Wermuth, Handbook of Pharmaceutical Salts: Properties, Selection, and Use [M], Wiley-VCH, 2002. Methods for preparing pharmaceutically acceptable salts of the compounds of the present invention are known to those skilled in the art.

[0092] The term "pharmaceutically acceptable ester" refers to an ester that is substantially non-toxic to an organism and is hydrolyzed in vivo to form a compound of the present invention or a salt thereof. Pharmaceutically acceptable esters generally include, but are not limited to, esters formed between a compound of the present invention and a pharmaceutically acceptable carboxylic acid or sulfonic acid, such esters being also referred to as carboxylate esters or sulfonate esters.

[0093] The term "isomers" refers to compounds that have the same number and types of atoms and therefore the same molecular weight, but differ in the arrangement or configuration of the atoms in space.

[0094] The term "stereoisomer" (or "optical isomer") refers to a stable isomer that has a perpendicular asymmetric plane due to at least one chiral factor (including a chiral center, chiral axis, chiral plane, etc.), thereby being able to rotate plane-polarized light. Because the compounds of the present invention may have asymmetric centers and other chemical structures that may lead to stereoisomerism, the present invention also includes these stereoisomers and mixtures thereof. Unless otherwise indicated, all stereoisomeric forms of the compounds of the present invention are within the scope of the present invention.

[0095] The term "tautomer" (or "tautomeric form") refers to structural isomers with different energies that can be interconverted via a low energy barrier. If tautomerism is possible (such as in solution), a chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (or prototropic tautomers) include, but are not limited to, interconversions via proton migration, such as keto-enol isomerization, imine-enamine isomerization, amide-iminoalcohol isomerization, and the like. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.

[0096] The term "solvate" refers to a substance formed by the combination of a compound of the present invention (or a pharmaceutically acceptable salt thereof) and at least one solvent molecule through non-covalent intermolecular forces. For example, solvates include, but are not limited to, hydrates (including hemihydrates, monohydrates, dihydrates, trihydrates, etc.), ethanolates, acetonides, etc.

[0097] The term "nitrogen oxide" refers to a compound formed by oxidation of a nitrogen atom in a tertiary amine or nitrogen-containing (aromatic) heterocyclic compound structure. For example, the nitrogen atom in the parent nucleus of the compound of formula I can form a corresponding nitrogen oxide.

[0098] The term "isotopically labeled" refers to a derivative compound formed by replacing a specific atom in a compound of the present invention with an isotope thereof. Unless otherwise indicated, the compounds of the present invention include various isotopes of H, C, N, O, F, P, S, and Cl, such as but not limited to 2 H(D), 3 H(T), 13 C. 14 C. 15 N. 17 O. 18 O. 18 F. 31 P. 32 P. 35 S. 36 S and 37 Cl.

[0099] The term "metabolite" refers to a derivative compound formed after metabolism of a compound of the present invention. Further information on metabolism can be found in Goodman and Gilman's: The Pharmacological Basis of Therapeutics (9 th ed.) [M], McGraw-Hill International Editions, 1996. The present invention encompasses all possible metabolites of the compounds of the invention, i.e., substances formed in the body of a subject to whom the compounds of the invention are administered. Metabolites of the compounds can be identified by techniques known in the art, and their activity can be characterized experimentally.

[0100] The term "prodrug" refers to a derivative compound that can directly or indirectly provide a compound of the present invention after being administered to an individual. Particularly preferred derivative compounds or prodrugs are compounds that can improve the bioavailability of the compound of the present invention when administered to an individual (e.g., more easily absorbed into the blood), or compounds that promote the delivery of the parent compound to the site of action (e.g., the lymphatic system). Unless otherwise indicated, all prodrug forms of the compounds of the present invention are within the scope of the present invention, and various prodrug forms are known in the art, for example, see T. Higuchi, V. Stella, Pro-drugs as Novel Drug Delivery Systems [J], American Chemical Society, Vol. 14, 1975. In addition, the present invention also encompasses compounds of the present invention containing protecting groups. In any process for preparing the compounds of the present invention, it may be necessary and / or desirable to protect sensitive groups or reactive groups on any related molecules, thereby forming a chemically protected form of the compound of the present invention. This can be achieved by conventional protecting groups, such as those described in TW Greene, PGM Wuts, Protective Groups in Organic Synthesis [M], John Wiley & Sons, 2006. These protecting groups may be removed at an appropriate subsequent stage using methods known in the art.

[0101] The term "independently" means that at least two groups (or ring systems) present in a structure with the same or similar range of values may have the same or different meanings in specific circumstances. For example, if substituent X and substituent Y are independently hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl, then when substituent X is hydrogen, substituent Y may be either hydrogen, or halogen, hydroxyl, cyano, alkyl, or aryl. Similarly, when substituent Y is hydrogen, substituent X may be either hydrogen, or halogen, hydroxyl, cyano, alkyl, or aryl.

[0102]

[0046] The term "halogen," as used herein, alone or in combination with other groups, refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0103] When used alone or in combination with other groups herein, the term "alkyl" refers to a straight-chain or branched aliphatic hydrocarbon group. For example, the term "C 1-6 The term "alkyl" refers to an alkyl group having 1 to 6 carbon atoms. For example, the alkyl group may be methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, or tert-butyl.

[0104] When used herein alone or in combination with other groups, the term "alkylene" refers to a straight-chain or branched divalent saturated aliphatic hydrocarbon group, and the two groups (or fragments) connected thereto may be connected to the same carbon atom or to different carbon atoms. For example, the term "C 1-6 The term "alkylene" refers to an alkylene group having 1 to 6 carbon atoms (e.g., methylene, 1,1-ethylene, 1,2-ethylene, 1,2-propylene, 1,3-butylene, etc.).

[0105] As used herein, the term "alkoxy" refers to an alkyl group attached to the rest of the molecule via an oxygen atom. For example, an alkoxy group can be methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, and the like.

[0106] When used herein alone or in combination with other groups, the term "cycloalkyl" refers to a saturated or partially saturated, monocyclic or polycyclic (such as bicyclic, for example, cyclic, bridged or spiro) non-aromatic hydrocarbon group. For example, the term "C 3-6 "Cycloalkyl" refers to a cycloalkyl group having 3 to 6 carbon atoms. For example, the cycloalkyl group can be cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or bicyclo[2.2.1]heptyl, etc. The cycloalkyl group in the present invention is optionally substituted with one or more substituents described herein.

[0107] The term "heterocyclyl" as used herein alone or in combination with other groups refers to a saturated or partially saturated, monocyclic or polycyclic (such as bicyclic, for example, fused, bridged or spiro) non-aromatic group, the ring atoms of which are composed of carbon atoms and at least one heteroatom selected from N, O and S, wherein the S atom is optionally substituted to form S(=O), S(=O)2 or S(=O)(=NR x ), R x Independently selected from H or C 1-4 Alkyl. If the valence bond requirements are met, the heterocyclic group can be connected to the rest of the molecule through any one of the ring atoms. For example, the term "3-8 membered heterocyclic group" as used in the present invention refers to a heterocyclic group having 3 to 8 ring atoms. For example, the heterocyclic group can be an oxirane group, an aziridine group, an azetidinyl group, an oxetane group, a tetrahydrofuranyl group, a dioxolyl group, a pyrrolidinyl group, a pyrrolidonyl group, an imidazolidinyl group, a pyrazolidinyl group, a tetrahydropyranyl group, a piperidinyl group, a piperazinyl group, a morpholinyl group, a thiomorpholinyl group, a dithianyl group, or a trithianyl group. The heterocyclic group in the present invention is optionally substituted with one or more substituents described herein.

[0108] When used herein alone or in combination with other groups, the term "aryl" refers to a monocyclic or fused polycyclic aromatic hydrocarbon group having a conjugated π electron system. For example, the term "C6-10 "Aryl" refers to an aromatic group having 6 to 10 carbon atoms. For example, the aromatic group can be phenyl, naphthyl, anthracenyl, phenanthrenyl, acenaphthenyl, azulenyl, fluorenyl, indenyl, pyrenyl, etc. The aromatic group in the present invention is optionally substituted with one or more substituents described herein.

[0109] As used herein, alone or in combination with other groups, the term "heteroaryl" refers to a monocyclic or fused polycyclic aromatic group having a conjugated π electron system, the ring atoms of which are composed of carbon atoms and at least one heteroatom selected from N, O and S. If the valence bond requirements are met, the heteroaryl group can be connected to the rest of the molecule through any one of the ring atoms. For example, the term "5-12 membered heteroaryl" as used in the present invention refers to a heteroaryl group having 5 to 12 ring atoms. For example, a heteroaryl group can be a thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl and benzo derivatives thereof, pyrrolopyridyl, pyrrolopyrazinyl, pyrazolopyridyl, imidazopyridyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl, purinyl, etc. The heteroaryl groups of the present invention are optionally substituted with one or more substituents as described herein (e.g., halogen, C 1-6 alkyl, etc.) substituted.

[0110]

[0046] The term "hydroxy," as used herein, alone or in combination with other groups, refers to -OH.

[0111]

[0046] The term "cyano," as used herein, alone or in combination with other groups, refers to -CN.

[0112] The term "amino," as used herein alone or in combination with other groups, refers to -NH2.

[0113]

[0046] The term "nitro," as used herein, alone or in combination with other groups, refers to -NO2. DETAILED DESCRIPTION

[0114] In order to make the purpose and technical solution of the present invention clearer, the embodiments of the present invention are described in detail below in conjunction with examples. However, it will be understood by those skilled in the art that the following examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention.

[0115] The reagents and instruments used in the examples are all commercially available conventional products. If no specific conditions are specified, the experiments were carried out under conventional conditions or the conditions recommended by the manufacturer. The term "room temperature" as used in the present invention refers to 20°C ± 5°C. When used to modify a certain numerical value or numerical range, the term "about" as used in the present invention refers to the numerical value or numerical range and the acceptable error range for those skilled in the art for the numerical value or numerical range, for example, the error range is ±10%, ±5%, ±4%, ±3%, ±2%, ±1%, ±0.5%, etc.

[0116] The structures of the compounds described in the following examples were confirmed by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS).

[0117] The nuclear magnetic resonance (NMR) was measured using a Bruker 400 MHz NMR instrument, with deuterated methanol (CD3OD), deuterated chloroform (CDCl3), hexadeuterated dimethyl sulfoxide (DMSO-d6) as the solvent, and tetramethylsilane (TMS) as the internal standard. 1 In H NMR, some hydrogen peaks may not appear due to interference from salts or solvents.

[0118] The abbreviations in the nuclear magnetic resonance (NMR) data in the following examples have the following meanings:

[0119] s: singlet, d: doublet, t: triplet, q: quartet, dd: double of doublet, qd: quadruple of doublet, ddd: double of doublet, ddt: double of doublets, dddd: double of doubles of doublets, m: multiplet, br: broad, J: coupling constant, Hz: Hertz, δ: chemical shift.

[0120] All chemical shift (δ) values are given in parts per million (ppm).

[0121] The mass spectrometry (MS) was performed using an Agilent 6120B mass spectrometer with an electrospray ionization (ESI) source.

[0122] HPLC analysis was performed using an Agilent 1200DAD high pressure liquid chromatograph (Sunfirc C18, 150×4.6 mm, 5 μm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18, 150×4.6 mm, 5 μm column).

[0123] The thin layer chromatography silica gel plate used was Qingdao Ocean GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) had a specification of 0.15mm-0.2mm, and the specification used for thin layer chromatography separation and purification products was 0.4mm-0.5mm silica gel plate.

[0124] Column chromatography generally uses Qingdao Ocean 200-300 mesh silica gel as the carrier.

[0125] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent systems used in the reactions were A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvents was adjusted according to the polarity of the compounds.

[0126] The eluent system for column chromatography and the developing solvent system for thin-layer chromatography used for purifying compounds include A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvents is adjusted according to the polarity of the compound, and a small amount of triethylamine and acidic or alkaline reagents can also be added for adjustment.

[0127] Synthesis of compounds

[0128] Synthesis Example 1: Synthesis of Intermediate M1a:

[0129]

[0130] Step a: The starting compound Ia (7.4 g, 50 mmol) and hydrazine hydrate (2.5 g, 50 mmol) were added to trifluoroacetic acid (100 mL), and the mixture was stirred at 120° C. for 2 hours. After the reaction was completed, the mixture was cooled to room temperature. A large amount of white solid precipitated, which was filtered. The filter cake was washed with plenty of water and then dried to obtain compound IIa (7.8 g, yield 96%). LCMS: [M+H] + =163. 1 H NMR (400MHz, DMSO-d6) δ11.55 (s, 2H), 8.25–7.99 (m, 2H), 7.88 (m, 2H).

[0131] Step b: Compound IIa (7.8 g, 48 mmol) was added to phosphorus oxychloride (50 mL), and the reaction was stirred at 110°C for 3 hours. After the reaction was completed, the temperature was cooled to room temperature and then slowly added to ice water. The pH was adjusted to 10 with 1N sodium hydroxide solution, and ethyl acetate (50 mL) was added. The organic phase was washed with saturated brine, and the combined organic phases were concentrated to give compound M1a (7.8 g, yield 67%), LCMS: [M+H]+=199. 1H NMR (400 MHz, DMSO-d6) δ8.48–8.16 (m, 4H).

[0132] Synthesis Example 2: Synthesis of Intermediate M2a:

[0133]

[0134] Step c: Add starting material IIIa (3.54 g, 20 mmol) and di-tert-butyl dicarbonate (5.45 g, 25 mmol) to tetrahydrofuran (THF) (50 mL) and heat under reflux for 2.5 hours. After completion of the reaction, the mixture was concentrated to obtain the crude product, which was then separated by column chromatography to afford M2a (5.15 g, 93% yield) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ9.70(s,1H),9.44(s,1H),7.46(s,2H),1.48(s,9H).

[0135] Example 1: Synthesis of Compound 1 (5-(((3,5-dichloro-4-((4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0136]

[0137] Synthesis method of compound 1:

[0138]

[0139] Step d: Intermediate M1a (2.5 g, 12.6 mmol), compound IIIa (2.24 g, 12.6 mmol), cuprous iodide (1.44 g, 7.56 mmol), and potassium carbonate (5.2 g, 37.8 mmol) were added to dimethyl sulfoxide (50 mL) and stirred at 40°C for 3 hours. After the reaction was completed, the reaction solution was poured into water (250 mL) to precipitate a large amount of solid, which was filtered and washed several times with methanol. The filter cake was dried to obtain a green solid compound IVa (4.26 g, yield 99%). LCMS: [M+H] + =340.

[0140] Step e: Compound IVa (1 g, 2.93 mmol), compound Va (980 mg, 11.7 mmol), sodium iodide (87.9 mg, 0.59 mmol), potassium carbonate (810 mg, 5.86 mmol) and acetonitrile (30 mL) were added to a sealed tube. After nitrogen replacement, the mixture was stirred at 100°C for 48 hours. After the reaction was completed, the mixture was concentrated, and ethyl acetate (50 mL) was added. The organic phase was washed with saturated brine, and the combined organic phases were concentrated to obtain a crude product. Column chromatography was then performed to obtain VIa (600 mg, 48% yield) as a white solid. LCMS: [M+H] + =426.

[0141] Step f: Compound VIa (600 mg, 1.41 mmol) and sodium acetate (579 mg, 7.06 mmol) were added to acetic acid (15 mL) and stirred at 110°C for 3 hours. After the reaction was completed, the mixture was concentrated, and ethyl acetate (50 mL) was added. The organic phase was washed with saturated sodium bicarbonate solution, and the combined organic phases were concentrated to obtain a crude product. Column chromatography was then performed to obtain a yellow solid VIIa (500 mg, yield 87%). LCMS: [M+H] + =408.

[0142] Step g: Compound VIIa (500 mg, 1.23 mmol) and hydrazine hydrate (5 mL) were added to ethanol (20 mL) and stirred at 90°C for 4 hours. After the reaction was completed, the mixture was concentrated to obtain a crude product, which was then separated by column chromatography to obtain a white solid, VIIIa (450 mg, 93% yield). LCMS: [M+H] + =394.

[0143] Step h: Compound VIIIa (450 mg, 1.14 mmol) and N,N'-disuccinimidyl carbonate (1.46 g, 5.7 mmol) were added to THF (10 mL) and stirred. Triethylamine (576 mg, 5.7 mmol) was then added and stirred at 80°C under nitrogen for 3 hours. After the reaction was complete, ethyl acetate (50 mL) was added and the organic phase was washed with saturated brine. The combined organic phases were concentrated to obtain the crude product. Column chromatography was then performed to obtain 1 (96 mg, 20% yield) as a white solid. LCMS: [M+H] + =420. 1 HNMR (400MHz, DMSO-d6) δ11.92(s,1H),8.27(d,1H),8.18(d,1H),8.00(m,2H),6.84(s,2H),6.77(t,J=6.5Hz,1H),4.27(d,J=6.2Hz,2H).

[0144] Example 2: Synthesis of Compound 2 (5-(((3,5-dimethyl-4-((4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0145]

[0146] Synthesis method of compound 2:

[0147]

[0148] The raw material IIIa was replaced with IIIb, and the synthesis method of compound 1 was referred to to obtain white solid compound 2 (20.2 mg, yield 22%). LCMS: [M+H] + =380. 1 H NMR(400MHz,DMSO-d6) δ12.21(s,1H),11.73(s,1H),8.25(d,J=7.6Hz,1H),8.21(d,J=7.6Hz,1H), 8.01(t,J=7.2Hz,1H),7.95(t,J=7.2Hz,1H),6.40(s,2H),6.05(t,J=6.4Hz, 1H), 4.19 (d, J = 6.4Hz, 2H), 1.99 (s, 6H).

[0149] Example 3: Synthesis of Compound 3 (5-(2-((3,5-dichloro-4-((4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)ethyl)-1,3,4-oxadiazol-2(3H)-one)

[0150]

[0151] Synthesis method of compound 3:

[0152]

[0153] Compound Va was replaced with Vb, and the yellow solid compound 3 (15.3 mg, yield 16%) was synthesized according to the synthesis method of compound 1. LCMS: [M+H] + =434. 1 H NMR(400MHz,DMSO-d6) δ12.09(s,1H),11.91(s,1H),8.27(d,J=7.6Hz,1H),8.19(d,J=7.6Hz,1H), 8.04(t,J=7.2Hz,1H),7.97(t,J=7.2Hz,1H),6.75(s,2H),6.35(t,J=5.6Hz, 1H), 3.42-3.37 (m, 2H), 2.77 (t, J = 6.0Hz, 2H).

[0154] Example 4: Synthesis of Compound 4 (5-(3-((3,5-dichloro-4-((4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)propyl)-1,3,4-oxadiazol-2(3H)-one)

[0155]

[0156] Synthesis method of compound 4:

[0157]

[0158] Compound Va was replaced with Vc, and the yellow solid compound 4 (62.4 mg, yield 45%) was synthesized according to the synthesis method of compound 1. LCMS: [M+H] + =448. 1 H NMR(400MHz,DMSO-d6) δ12.04(s,1H),11.90(s,1H),8.25-8.23(m,1H),8.17-8.14(m,1H),8.03-7 .99(m,1H),7.97-7.93(m,1H),6.68(s,2H),6.26(t,J=5.6Hz,1H),3.07(q, J=6.4Hz,2H),2.61(t,J=7.6Hz,2H),1.84-1.77(m,2H).

[0159] Example 5: Synthesis of Compound 5 (N-(3,5-dichloro-4-((4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)-5-oxo-4,5-dihydro-1,3,4-oxadiazole-2-carboxamide)

[0160]

[0161] Synthesis method of compound 5:

[0162]

[0163] Compound Va was replaced with Vd, and the synthesis method of compound 1 was referred to obtain white solid compound 5 (29 mg, yield 27%). LCMS: [M+H] + =434. 1 H NMR (400MHz, DMSO-d6) δ12.01(s,1H),10.21(s,1H),8.32–8.28(m,1H),8.25–8.21(m,1H),8.10–7.99(m,4H).

[0164] Example 6: Synthesis of Compound 6 (N-(3,5-dichloro-4-((4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)-2-(5-oxo-4,5-dihydro-1,3,4-oxadiazol-2-yl)acetamide

[0165]

[0166] Synthesis method of compound 6:

[0167]

[0168] Compound Va was replaced with Ve, and the yellow solid compound 6 (5.2 mg, yield 3.9%) was synthesized according to the synthesis method of compound 1. LCMS: [M+H] + =448. 1 H NMR(400MHz,DMSO-d6) δ11.98(s,1H),10.72(s,1H),8.29(d,J=8.0Hz,1H),8.21(d,J=8.0Hz,1H ), 8.06 (t, J = 7.6Hz, 1H), 8.00 (t, J = 7.6Hz, 1H), 7.79 (s, 2H), 3.80 (s, 2H).

[0169] Example 7: Synthesis of Compound 7 (5-(((3,5-dichloro-4-((5-oxo-5,6-dihydropyrido[2,3-d]pyridazin-8-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0170]

[0171] Synthesis method of compound 7:

[0172]

[0173] Compound M1a was replaced with M1b, and the synthesis method of compound 1 was referred to obtain white solid compound 7 (51.9 mg, yield 41%), LCMS: [M+H] + =421. 1 H NMR (400MHz, DMSO-d6) δ12.17(s,1H),9.16(dd,J=4.4,1.6Hz,1H),8.59(dd,J=8.4,1.6 Hz, 1H), 7.99 (dd, J = 8.4, 4.4Hz, 1H), 6.83 (s, 2H), 6.79 (t, J = 6.4Hz, 1H), 4.29 (d, J = 6.4Hz, 2H).

[0174] Example 8: Synthesis of Compound 8 (5-(((3,5-dichloro-4-((4-oxo-3,4,5,6,7,8-hexahydrophthalazin-1-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0175]

[0176] Synthesis method of compound 8:

[0177]

[0178] Compound M1a was replaced with M1c, and the synthesis method of compound 1 was referred to obtain white solid compound 8 (38 mg, yield 35%). LCMS: [M+H] + =424. 1 H NMR (400MHz, DMSO-d6) δ11.95(s,1H),6.80(s,2H),6.74(t,J=6.4Hz,1H),4.28(d,J=6.4 Hz, 2H), 2.63–2.54 (m, 2H), 2.45–2.32 (m, 2H), 1.79–1.60 (m, J = 4.8Hz, 4H).

[0179] Example 9: Synthesis of Compound 9 (5-(((3,5-dichloro-4-((7,7-dimethyl-1-oxo-2,5,6,7-tetrahydro-1H-cyclopenta[d]pyridazin-4-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0180]

[0181] Synthesis method of compound 9:

[0182]

[0183] Compound M1a was replaced with M1d, and the synthesis method of compound 1 was referred to obtain white solid compound 9 (32 mg, yield 29%). LCMS: [M+H] + =438. 1 H NMR (400MHz, DMSO-d6) δ11.94(s,1H),6.81(s,2H),6.75(t,J=6.0Hz,1H),4.27(d,J=5.6 Hz, 2H), 2.88 (t, J = 6.8 Hz, 2H), 1.95 (t, J = 7.2 Hz, 2H), 1.32 (s, 4H).

[0184] Example 10: Synthesis of Compound 10 (5-(((3,5-dichloro-4-((4-oxo-3,4,5,6,7,8-hexahydro-5,8-methanophthalazin-1-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0185]

[0186] Synthesis method of compound 10:

[0187]

[0188] Compound M1a was replaced with M1e, and the synthesis method of compound 1 was referred to obtain white solid compound 10 (12.6 mg, yield 13%). LCMS: [M+H] + =436. 1 H NMR (400MHz, DMSO-d6) δ11.97(s,1H),6.79(s,2H),6.74(t,J=6.0Hz,1H),4.28(d,J=5.6 Hz,2H),2.10–1.86(m,3H),1.71(d,J=8.8Hz,1H),1.47(d,J=8.8Hz,1H),1.22(s,1H),1.19–1.12(m,1H),1.08–0.96(m,1H).

[0189] Example 11: Synthesis of Compound 11 (5-(1-((3,5-dichloro-4-((4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)ethyl)-1,3,4-oxadiazol-2(3H)-one)

[0190]

[0191] Synthesis method of compound 11:

[0192]

[0193] Compound Va was replaced with Vf, and the synthesis method of compound 1 was referred to obtain white solid compound 11 (43 mg, yield 29%). LCMS: [M+H] + =434. 1 H NMR(400MHz,DMSO-d6) δ8.31(d,J=7.8Hz,1H),8.17(d,J=7.8Hz,1H),8.08–7.96(m,2H),6.85(s,2H),6.69(d,J=8.2Hz,1H),4.67(p,J=7.0Hz,1H),1.47(d,J=6.8Hz, 3H).

[0194] Example 12: Synthesis of Compound 12 (5-(((3,5-dichloro-4-((3-methyl-4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0195]

[0196] Synthesis method of compound 12:

[0197]

[0198] Step i: Referring to the synthetic route of compound 1, intermediate XIa (350 mg, 0.69 mmol) was synthesized. The intermediate XIa and potassium carbonate (476 mg, 3.45 mmol) were added to N,N-dimethylformamide (DMF) (10 mL) and stirred. Then, iodomethane (129 mg, 0.83 mmol) was added thereto and the reaction was stirred at room temperature for 3 hours. After the reaction was completed, ethyl acetate (92 mL) was added, and the organic phase was washed with saturated brine. The organic phases were combined and concentrated to obtain a crude product. Then, the colorless oil XIIa (353 mg, yield 98%) was obtained by column chromatography. LCMS: [M+H] + =522.

[0199] Step j: Compound XIIa (353 mg, 0.68 mmol) was added to a mixed solvent of trifluoroacetic acid (3 mL) and dichloromethane (10 mL) and stirred at room temperature overnight. After the reaction was completed, dichloromethane (50 mL) was added, and the organic phase was washed with saturated sodium bicarbonate solution. The combined organic phases were concentrated to obtain a crude product. Column chromatography was then performed to obtain a yellow solid XIIIa (282 mg, yield 99%). LCMS: [M+H] + =422.

[0200] Step g: Compound XIIIa (282 mg, 0.67 mmol) and hydrazine hydrate (3 mL) were added to ethanol (10 mL) and stirred at 90°C for 3 hours. After the reaction was completed, the mixture was filtered and the filter cake was dried to obtain a white solid XIVa (120 mg, yield 44%). LCMS: [M+H] + =408.

[0201] Step h: Compound XIVa (120 mg, 0.3 mmol) and N,N'-disuccinimidyl carbonate (230.4 mg, 0.9 mmol) were added to THF (5 mL) and stirred. Triethylamine (91 mg, 0.9 mmol) was then added and stirred at 90°C under nitrogen for 3 hours. After the reaction was complete, ethyl acetate (10 mL) was added and the organic phase was washed with saturated brine. The combined organic phases were concentrated to obtain the crude product. Column chromatography was then performed to obtain 12 (27.4 mg, 21.1% yield) as a white solid. LCMS: [M+H] + =434. 1 H NMR(400 MHz, DMSO-d6)δ8.30(d,J=8.0Hz,1H),8.19(d,J=8.0Hz,1H),8.08–7.94(m,2H),6.85(s,2H),6.80(t,J=6.0Hz,1H),4.31(d,J=6.0Hz,2H),3.45(s, 3H).

[0202] Example 13: Synthesis of Compound 13 (5-(((3,5-dichloro-4-((3-(methyl-d3)-4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0203]

[0204] Synthesis method of compound 13:

[0205]

[0206] Iodomethane was replaced with deuterated iodomethane, and the synthesis method of compound 12 was referred to obtain white solid compound 13 (72 mg, yield 30%), LCMS: [M+H] + =437. 1 H NMR(400MHz, DMSO-d6)δ12.22(s,1H),8.30(d,J=8.4Hz,1H),8.19(d,J=8.0Hz,1H), 8.07–7.95(m,2H),6.85(s,2H),6.80(t,J=6.4Hz,1H),4.31(d,J=6.4Hz,2H).

[0207] Example 14: Synthesis of Compound 14 (5-(1-((3,5-dichloro-4-((3-methyl-4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)ethyl)-1,3,4-oxadiazol-2(3H)-one)

[0208]

[0209] Synthesis method of compound 14:

[0210]

[0211] Compound Va was replaced with compound Vf, and the synthesis method of compound 12 was referred to obtain white solid compound 14 (41.4 mg, yield 24%). LCMS: [M+H] + =448. 1 H NMR(400MHz, DMSO-d6)δ8.30(d,J=7.6Hz,1H),8.18(d,J=7.6Hz,1H),8.07–7.96(m, 2H), 6.85 (s, 2H), 6.69 (d, J = 8.0Hz, 1H), 4.68 (p, J = 7.2Hz, 1H), 3.45 (s, 3H), 1.46 (d, J = 6.8Hz, 3H).

[0212] Example 15: Synthesis of Compound 15 (5-(((3,5-dichloro-4-((3-ethyl-4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0213]

[0214] Synthesis method of compound 15:

[0215]

[0216] Iodomethane was replaced with iodoethane, and the synthesis method of compound 12 was referred to obtain white solid compound 15 (27.4 mg, yield 22%). LCMS: [M+H] + =448. 1 H NMR(400MHz, DMSO-d6)δ8.31(d,J=8.2Hz,1H),8.18(d,J=7.2Hz,1H),8.05–7.97(m, 2H), 6.86 (s, 2H), 6.79 (t, J = 6.4Hz, 1H), 4.31 (d, J = 6.2Hz, 2H), 3.87 (t, J = 7.2Hz, 2H), 1.10 (d, J = 7.2Hz, 3H).

[0217] Example 16: Synthesis of Compound 16 (5-(((3,5-dichloro-4-((4-oxo-3-propyl-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0218]

[0219] Synthesis method of compound 16:

[0220]

[0221] Iodomethane was replaced with 1-iodopropane, and the synthesis method of compound 12 was referred to obtain white solid compound 16 (77.2 mg, yield 36%). LCMS: [M+H] + =462. 1H NMR(400MHz, DMSO-d6)δ12.26(s,1H),8.31(d,J=7.8Hz,1H),8.19(d,J=7.8Hz,1H), 8.01(dt,J=19.4,7.4Hz,2H),6.87(s,2H),6.78(t,J=6.4Hz,1H),4.32(d,J =6.2Hz,2H),3.83(t,J=6.8Hz,2H),1.55(q,J=7.2Hz,2H),0.75(t,J=7.4Hz, 3H).

[0222] Example 17: Synthesis of Compound 17 (5-(((3,5-dichloro-4-((3-(2-fluoroethyl)-4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0223]

[0224] Synthesis method of compound 17:

[0225]

[0226] Iodomethane was replaced with 1-fluoro-2-iodoethane, and the synthesis method of compound 12 was referred to obtain white solid compound 17 (34.3 mg, yield 15%). LCMS: [M+H] + =466. 1 H NMR(400MHz, DMSO-d6)δ12.26(s,1H),8.33(d,J=7.8Hz,1H),8.21(d,J=7.8Hz,1H), 8.04(dt,J=20.2,7.4Hz,2H),6.87(s,2H),6.79(t,J=6.4Hz,1H),4.57(dt ,J=47.4,4.8Hz,2H), 4.32(d,J=6.2Hz,2H), 4.20(dt,J=26.4,4.8Hz,2H).

[0227] Example 18: Synthesis of Compound 18 (5-(((4-((3-butyl-4-oxo-3,4-dihydrophthalazin-1-yl)oxy)-3,5-dichlorophenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0228]

[0229] Synthesis method of compound 18:

[0230]

[0231] Iodomethane was replaced with 1-iodobutane, and the synthesis method of compound 12 was referred to obtain white solid compound 18 (89.6 mg, yield 41%). LCMS: [M+H] + =476. 1 H NMR(400MHz, DMSO-d6)δ12.26(s,1H),8.31(d,J=7.4Hz,1H),8.19(d,J=8.6Hz,1H), 8.12–7.95(m,2H),6.87(s,2H),6.80(t,J=6.4Hz,1H),4.32(d,J=6.4Hz,2H),3.87(t,J=6.6Hz,2H),1.52(p,J=6.8Hz,2H),1.15(h,J=7.4Hz,2H), 0.76(s,3H).

[0232] Example 19: Synthesis of Compound 19 (5-(((3,5-dichloro-4-((3-isopropyl-4-oxo-3,4-dihydrophthalazin-1-yl)oxy)phenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0233]

[0234] Synthesis method of compound 19:

[0235]

[0236] Iodomethane was replaced with 2-iodopropane, and the synthesis method of compound 12 was referred to obtain white solid compound 19 (26.8 mg, yield 18%). LCMS: [M+H] + =462. 1 H NMR(400MHz, DMSO-d6)δ12.25(s,1H),8.31(d,J=8.2Hz,1H),8.18(d,J=7.8Hz,1H), 8.11–7.95(m,2H),6.88(s,2H),6.75(t,J=6.4Hz,1H),5.13–5.02(m,1H),4.32(d,J=6.4Hz,2H),1.01(d,J=6.6Hz,6H).

[0237] Example 20: Synthesis of Compound 20 (5-(((4-((3-benzyl-4-oxo-3,4-dihydrophthalazin-1-yl)oxy)-3,5-dichlorophenyl)amino)methyl)-1,3,4-oxadiazol-2(3H)-one)

[0238]

[0239] Synthesis method of compound 20:

[0240]

[0241] Iodomethane was replaced with benzyl iodide, and the synthesis method of compound 12 was referred to obtain white solid compound 20 (68 mg, yield 31%), LCMS: [M+H] + =510. 1 H NMR(400MHz,DMSO-d6) δ12.23(s,1H),8.27(d,J=8.4Hz,1H),8.17(d,J=7.8Hz,1H),8.03–7.94(m ,2H),7.22–7.18(m,3H),7.13–7.10(m,2H),6.82(s,2H),6.77(t,J=6.4Hz, 1H), 4.98 (s, 2H), 4.28 (d, J = 3.2Hz, 2H).

[0242] Pharmacological activity test

[0243] Experimental Example 1: Detection of Compounds' Agonistic Activity on THRα / β Based on Time-Resolved Fluorescence Resonance Energy Transfer (THR-FRET)

[0244] 1. Construction of THRα / β overexpression vector

[0245] The THRα / β LBD domain sequence was found by consulting NCBI, and the pET21-His-GST-dLBT-THRαLBD and pET21-His-GST-dLBT-THRβ LBD overexpression vectors were constructed by fusion method, and the accuracy of the sequences was confirmed by sequencing.

[0246] 2. Prokaryotic expression of recombinant proteins in Escherichia coli

[0247] The correctly sequenced THRαLBD and THRβLBD overexpression vectors were transformed into Escherichia coli BL21(DE3) cells and plated on ampicillin-resistant agar plates. Single colonies were picked and expanded in LB medium and transferred to 1L of LB at a 1:100 ratio for large-scale culture. When the OD value reached 0.8-1.2, 0.5mM isopropyl-β-D-thiogalactopyranoside (IPTG) was added and induced overnight at 18°C. The cells were harvested, disrupted, and purified using a GST column and molecular sieves to obtain His-GST-dLBT-THRαLBD and His-GST-dLBT-THRβLBD proteins. Protein concentrations were determined using a Sangon Bradford protein quantification kit to be 24μM and 23μM, respectively.

[0248] 3. Compound Preparation and Reaction System Preparation

[0249] The protein was taken out of the -80 degree freezer, the GST-tagged THRα / βLBD domain protein and Eu-labeled GST antibody were slowly thawed on ice, and a detection buffer containing a final concentration of 5 mM dithiothreitol (DTT) was prepared.

[0250] 3.1 Compound preparation

[0251] The starting concentration of the compound was 100 μM (in DMSO). The compound (100 μM in DMSO) was diluted 3-fold in DMSO to obtain 11 isocratic concentrations. The isocratic concentrations were then diluted 50-fold in a test solution containing 5 mM DTT.

[0252] 3.2 Preparation of THR-FRET reaction system

[0253] The final concentrations of all components were calculated based on a final volume of 20 μL per well. GST-tagged THRα / β protein, SRC2 (LKEKHKILHRLLQDSSSPV) polypeptide, XL665 (Cisobio, #610SAXLB), and Eu-labeled GST antibody were added to 18 μL of detection buffer containing 5 mM DTT. The final concentrations were 2 nM, 200 nM, 0.05 nM, and 7.6 nM, respectively, to prepare a reaction mixture of protein, polypeptide, and antibody in 18 μL per well.

[0254] 18 μL of reaction mixture and 2 μL of diluted compound were added to the optiplate-384-well plate and reacted at room temperature for 24 hours.

[0255] 3.3 Plate Reading

[0256] Plates were read using an MD i3X multifunctional microplate reader with excitation and emission wavelengths of 340 nm and 665 nm, respectively. The intensity of 616 nm light, generated by europium excitation with the MDi3X multifunctional microplate reader, was used as background. Depending on the degree of THRα and THRβ activation by different compounds, the intensity of 665 nm emission light generated by XL665 excitation at 616 nm varied. The intensity ratio of these two wavelengths (665 nm, 616 nm) was used as the compound's activation activity for THRα or THRβ, and the ratio was normalized to the solvent DMSO group. GraphPad Prism 6.0 software was used to fit the dose-response curve using four parameters to calculate the EC50 value.

[0257] 4. Results

[0258] Experimental data show that the compound of the present invention has strong THRβ agonist activity and certain THRα / β selectivity. Specific data are shown in Table 1.

[0259] Table 1

[0260]

[0261] *: 100μM≥EC50≥10μM; **: 10μM>EC50>5μM; ***: 5μM≥EC50

[0262] THRα / β≥5; 5>THRα / β>1; 1≥THRα / β

[0263] Experimental Example 2: Detection of the THRα and THRβ agonist activity of compounds based on reporter gene activity assay

[0264] 1. Methods

[0265] 1.1 Construction and preparation of plasmids pGAL4-FXR-LBD and pG5-Luc

[0266] The pGAL4-THRα-LBD and pGAL4-THRβ-LBD plasmids used in the reporter gene assay system were constructed using conventional molecular cloning methods. The main steps were: The cDNA sequences of THRα (NM_003250) and THRβ (NM_000461), corresponding to the amino acid sequences of THRα (163-407AA) and THRβ (217-461AA), were inserted into the BamHI and NotI restriction sites of the pGAL4 vector, respectively, using PCR to generate the pGAL4-THRα-LBD and pGAL4-THRβ-LBD plasmids. The pG5-Luc (#E249A) and pRL-TK (#E2241) plasmids were purchased from Promega. The plasmids were transformed into DH5α Escherichia coli using the CaCl2 method, cultured and amplified, and then purified using a plasmid extraction kit (TIANGEN, #D107).

[0267] 1.2 Plasmid co-transfection into HEK293T cells and compound treatment

[0268] One day before plasmid transfection, HEK293T cells were plated at 1×10 4 The density of cells / well was seeded in 96-well plates. HD (Promega, #E2311) instructions for cell transfection. The main steps are as follows: Taking one well as an example, add 20 ng, 50 ng and 5 ng of plasmids pGAL4-THRα-LBD or pGAL4-THRβ-LBD, pG5-Luc and pRL-TK to 10 uL of Opti-MEM TM I culture medium (Gibco, #11058021) and mix well; then add 0.25uL of HD, mix thoroughly, and let stand at room temperature for 5 minutes; then add 10 μL of this mixture to the cell well containing 100 μL of culture medium. Six hours after cell co-transfection, the compound was diluted three-fold with dimethyl sulfoxide at a maximum concentration of 1 μM. A total of 10 concentrations were added to the cell culture medium and treated for 24 hours in duplicate wells. Triiodothyronine (T3) was used as a positive control.

[0269] 1.3 Dual-Glo Luciferase Assay

[0270] After cells were treated with the compounds for 24 h, The Luciferase Assay System (Promega, #E2940) was used for detection. The main steps were: aspirate 50uL of culture medium from each well, then add 50uL Luciferase reagent, shake at room temperature for 10 minutes; take 80uL of the cleavage reaction solution to a white opaque optiPlate-96 well plate, and use an MD i3x multifunctional microplate reader to detect the luminescent signal value of firefly luciferase (Firefly-Luc); then add 40uL Stop& The reagent was added and shaken at room temperature for 10 minutes. The luminescence signal of Renilla luciferase (Renilla-Luc) was then measured using an MD i3x multi-function microplate reader. The ratio of Firefly-Luc / Renilla-Luc was used as the THR activation activity of the compound and normalized to the ratio of the solvent group (DMSO). GraphPad Prism 6.0 software was used to fit the dose-response curve using a four-parameter method to calculate the EC50 value.

[0271] 2. Results

[0272] Experimental data show that the compound of the present invention has strong THRβ agonist activity and certain THRα / β selectivity. Specific data are shown in Table 2.

[0273] Table 2

[0274]

[0275] *:100μM≥EC50≥10μM;**:10μM>EC50>5μM;***:5μM≥EC50

[0276] THRα / β≥5; 5>THRα / β>1; 1≥THRα / β.

Claims

1. A compound having the structure of formula (1) or a pharmaceutically acceptable salt thereof: in, Ring A is selected from phenyl, cyclopentyl, cyclohexyl or bicyclo[2.2.1]heptyl, said ring A being optionally substituted by one or more R4, said R4 being independently selected from halogen, cyano, amino, nitro, hydroxyl, C 1-6 Alkyl or C 1-6 Alkoxy, R1 is selected from hydrogen or C 1-6 Alkyl, said alkyl being optionally substituted by one or more independently selected from deuterium, halogen, cyano, amino, nitro, hydroxyl, C 1-6 Alkyl or C 1-6 Alkoxy substituents are substituted, R2 and R3 are independently selected from hydrogen, halogen, cyano, amino, nitro, hydroxy, C 1-6 Alkyl or C 1-6 Alkoxy, X is selected from -C 1-6 Alkylene-, the alkylene is optionally substituted by one or more independently selected from halogen, cyano, amino, nitro, hydroxyl, C 1-6 Alkyl or C 1-6 The substituents of the alkoxy group are substituted.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Ring A is selected from:

3. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R1 is selected from hydrogen, methyl, -CD3, ethyl, 4. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R2 and R3 are independently selected from Cl, Br or methyl.

5. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein X is selected from -CH2-, -CH2CH2-, -CH2CH2CH2- or -CH(CH3)-.

6. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

7. A pharmaceutical composition comprising the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

8. Use of the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 7, in the preparation of a medicament for treating non-alcoholic fatty liver disease, dyslipidemia, atherosclerosis or hypothyroidism mediated by thyroid hormone beta receptor.

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