1,2,4-triazolone derivatives as dhodh inhibitors, processes for their preparation and uses thereof

Abstract

Provided are 1,2,4-triazolone derivatives as DHODH inhibitors, and preparation methods and uses thereof. Specifically, provided are compounds represented by general formula (1) and preparation methods thereof, and uses of the compounds represented by general formula (1) and isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates thereof as DHODH inhibitors in the preparation of drugs for treating DHODH-related diseases.

Description

[0001] This application claims priority to Chinese patent application 202110757483X, filed on July 5, 2021. The entire contents of the aforementioned Chinese patent application are incorporated herein by reference. Technical Field

[0002] This invention relates to the field of medicinal chemistry, and more specifically, to a class of novel 1,2,4-triazolone derivatives, their preparation methods, and the uses of such compounds. Background Technology

[0003] Dihydroorotate dehydrogenase (DHODH) is an iron-containing flavin-dependent enzyme primarily located on the inner mitochondrial membrane. It is a key enzyme in the de novo synthesis of pyrimidine nucleotides, which are structural components of DNA and RNA. Orotic acid is a precursor in the synthesis of pyrimidine nucleotides. DHODH catalyzes the dehydrogenation of dihydroorotate, converting it to orotic acid. Therefore, inhibiting DHODH can block the de novo synthesis of pyrimidines, leading to impaired DNA and RNA synthesis.

[0004] Intracellular pyrimidine nucleotides primarily originate from de novo and salvage pathways. In normally differentiated resting cells, pyrimidine nucleotides mainly come from the salvage pathway, also known as the regenerative pathway, which directly converts free pyrimidine bases into pyrimidine nucleotides. However, for metabolically active T and B lymphocytes or rapidly dividing cancer cells under pathological conditions, the pyrimidine nucleotides generated by the salvage pathway are insufficient to maintain cell function or survival. Therefore, de novo synthesis of pyrimidine nucleotides dependent on DHODH is crucial for the survival of these cells. Inhibiting DHODH activity can selectively prevent these cells from meeting their high demand for pyrimidine nucleotides, thereby hindering the synthesis of biomolecules such as DNA, RNA, and glycoproteins, and suppressing cell function or growth.

[0005] Leveraging this property, the development of DHODH inhibitors targeting abnormally activated T and B cells, as well as rapidly dividing cancer cells, for the treatment of cancer, viral infections, and autoimmune diseases (rheumatoid arthritis and multiple sclerosis) has shown initial success. For example, the small molecule DHODH inhibitors leflunomide / teriflunomide and bukquina have been approved for the treatment of rheumatoid arthritis and multiple sclerosis. However, these inhibitors are highly lipid-soluble and have poor selectivity, leading to significant toxic side effects with long-term use. Therefore, the search for novel, highly effective DHODH inhibitors with fewer side effects for the treatment of immune-related diseases and tumors has become a current research hotspot. Summary of the Invention

[0006] This invention provides a compound of general formula (1) or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates:

[0007]

[0008] In general formula (1):

[0009] n is 0, 1, or 2;

[0010] X is CH or N;

[0011] R 1 It is an aryl or heteroaryl group, and the aryl and heteroaryl groups may be substituted by one or more of the following groups: halogen, NH2, C1-C3 alkyl, C1-C3 alkoxy or halo-C1-C3 alkyl;

[0012] R 2 For H or F;

[0013] R 3 It is a C1-C3 alkyl group;

[0014] R 4 It is a C1-C6 alkyl, C3-C8 cycloalkyl, or halogenated C1-C6 alkyl.

[0015] In another preferred embodiment, in the general formula (1), R 1 for: Where R a R b R c R d and R e It can be H, F, Cl, Me, Et, OMe, CHF2, CF3 or NH2 independently.

[0016] In another preferred embodiment, in the general formula (1), R 3 For Me.

[0017] In another preferred embodiment, in the general formula (1), R 4 For Me, Et, CF3,

[0018] In some embodiments of the present invention, the above-mentioned compound or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates are selected from one of the following structures:

[0019]

[0020]

[0021]

[0022]

[0023] Another object of the present invention is to provide a pharmaceutical composition comprising a pharmaceutically acceptable carrier, a diluent and / or an excipient, and a compound of general formula (1) of the present invention, or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates as active ingredients.

[0024] Another object of the present invention is to provide the use of the compound of general formula (1) of the present invention, or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates, or the above-described pharmaceutical compositions, in the preparation of a medicament for treating tumors and related diseases; wherein the tumor is preferably a solid tumor and a hematologic tumor.

[0025] It should be understood that the foregoing general description of the invention and the following detailed description are exemplary and illustrative, and are intended to provide further explanation of the claimed invention.

[0026] Compound Synthesis

[0027] The preparation method of the compound of general formula (1) of the present invention is described in detail below, but these specific methods do not constitute any limitation on the present invention.

[0028] The compounds of general formula (1) described above can be synthesized using standard synthetic techniques or known techniques combined with the methods described herein. Furthermore, the solvents, temperatures, and other reaction conditions mentioned herein can be varied. Starting materials used for the synthesis of the compounds can be obtained synthetically or from commercial sources. The compounds described herein and other related compounds with different substituents can be synthesized using known techniques and starting materials, including those discovered in March, A DVANCED O RGANIC C HEMISTRY 4 th Ed., (Wiley 1992); Carey and Sundberg, A DVANCED O RGANIC C HEMISTRY 4 th Ed., Vols. A and B (Plenum 2000, 2001), Green and Wuts, P ROTECTIVE G ROUPS IN O RGANIC S YNTHESIS 3 rd The method described in Ed. (Wiley 1999) can be altered by using appropriate reagents and conditions that introduce different groups into the molecular formula provided herein.

[0029] On the one hand, the compounds described herein are prepared according to methods known in the art. However, the conditions of the method, such as reactants, solvents, bases, amounts of compounds used, reaction temperatures, and reaction times, are not limited to the explanations below. The compounds of the present invention can also be conveniently prepared by combining various synthetic methods described in this specification or known in the art, such combinations being readily performed by those skilled in the art. On the other hand, the present invention also provides a method for preparing the compound of general formula (1), wherein the compound of general formula (1) can be prepared by the following method A or method B:

[0030] Method A includes the following steps: First, compounds A1 and A2 react under strongly alkaline conditions to generate compound A3. Compound A3 further reacts with compound A4 under strongly alkaline conditions to generate compound A5. Compound A5 and compound A6 undergo a coupling reaction to generate the target compound A7.

[0031]

[0032] In the above reaction equation, n and R 1 R 2 R 3 and R 4 The definition is as described above.

[0033] Method B comprises the following steps: First, the carboxyl group of compound B1 undergoes a chlorination reaction to generate acyl chloride compound B2. Compound B2 further reacts with isopropanol under strong alkaline conditions to generate compound B3. Compound B3 and compound A6 react under alkaline conditions to generate compound B4. After compound B4 reacts under strong alkaline conditions, it reacts with compound A2 to obtain compound B5. Compound B5 reacts with a suitable starting material A4 to obtain the target compound B6.

[0034]

[0035] In the above reaction equation, n and R 1 R 2 R 3 and R 4 The definition is as described above.

[0036] Further forms of the compound

[0037] "Pharmaceutical acceptable" here means that a substance, such as a carrier or diluent, will not destroy the biological activity or properties of a compound and is relatively non-toxic. For example, when given to an individual, a substance will not cause unwanted biological effects or interact with any of its components in a harmful manner.

[0038] The term "pharmaceutically acceptable salt" refers to a form of a compound that does not cause significant irritation to the administered organism and does not diminish the compound's biological activity and properties. In some specific respects, pharmaceutically acceptable salts are obtained by reacting compounds of general formula (1) with acids, such as inorganic acids like hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, and carbonic acid; organic acids like formic acid, acetic acid, propionic acid, oxalic acid, trifluoroacetic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid; and acidic amino acids like aspartic acid and glutamic acid.

[0039] It should be understood that references to pharmaceutically acceptable salts include solvent-added forms or crystalline forms, especially solvates or polymorphs. Solvates contain stoichiometric or non-stoichiometric solvents and are selectively formed during crystallization with pharmaceutically acceptable solvents such as water, ethanol, etc. A hydrate is formed when the solvent is water, or an alcohol is formed when the solvent is ethanol. Solvates of compounds of general formula (1) are readily prepared or formed according to the methods described herein. For example, hydrates of compounds of general formula (1) are readily prepared by recrystallization from a mixture of water and organic solvents, including but not limited to tetrahydrofuran, acetone, ethanol, or methanol. Furthermore, the compounds mentioned herein can exist in both solvated and non-solvated forms. In summary, for the purposes of the compounds and methods provided herein, the solvated form is considered equivalent to the non-solvated form.

[0040] In other specific embodiments, compounds of general formula (1) are prepared in various forms, including but not limited to amorphous, pulverized, and nano-particle forms. Furthermore, compounds of general formula (1) include crystalline forms and can also be polymorphic. Polymorphs comprise different lattice arrangements of the same elemental composition of the compound. Polymorphs typically have different X-ray diffraction spectra, infrared spectra, melting points, densities, hardness, crystal forms, optical and electrical properties, stability, and solubility. Different factors such as recrystallization solvents, crystallization rates, and storage temperatures may cause a single crystal form to dominate.

[0041] In another aspect, compounds of general formula (1) may possess a chiral center and / or axial chirality, and thus appear as racemates, racemic mixtures, single enantiomers, diastereomers, and single diastereomers, and cis-trans isomers. Each chiral center or axial chirality will independently produce two optical isomers, and all possible optical isomers and diastereomer mixtures, as well as pure or partially pure compounds, are included within the scope of this invention. This invention means including all such isomeric forms of these compounds.

[0042] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium. 3 H), Iodine-125 ( 125 I) and C-14 14 C). For example, deuterium can be used to replace hydrogen atoms to form deuterated compounds. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs generally have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug half-life in vivo. All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0043] the term

[0044] Unless otherwise specified, the terms used in this application, including the specification and claims, are defined as follows. It must be noted that in the specification and appended claims, unless otherwise clearly indicated, the singular form "a" includes the plural meaning. Unless otherwise specified, conventional methods such as mass spectrometry, nuclear magnetic resonance, HPLC, protein chemistry, biochemistry, recombinant DNA techniques, and pharmacology are used. In this application, unless otherwise specified, "or" or "and" refers to "and / or".

[0045] Unless otherwise specified, "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched groups with 1 to 6 carbon atoms. Lower alkyl groups containing 1 to 4 carbon atoms are preferred, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, and tert-butyl. As used herein, "alkyl" includes unsubstituted and substituted alkyl groups, especially alkyl groups substituted with one or more halogens. Preferred alkyl groups are selected from CH3, CH3CH2, CF3, CHF2, CF3CH2, CF3(CH3)CH, and so on. n Pr, i Bu、 n Bu or t Bu.

[0046] Unless otherwise specified, "cycloalkyl" refers to a non-aromatic hydrocarbon ring system (monocyclic, bicyclic, or polycyclic). A partially unsaturated cycloalkyl group may be referred to as "cycloalkenyl" if the carbon ring contains at least one double bond, or as "cycloynyl" if the carbon ring contains at least one triple bond. Cycloalkyl groups may include monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings) groups and spirocyclic groups. In some embodiments, the cycloalkyl group is monocyclic. In some embodiments, the cycloalkyl group is monocyclic or bicyclic. The cycloforming carbon atom of the cycloalkyl group may optionally be oxidized to form an oxo or thio group. Cycloalkyl groups also include cycloalkylene groups. In some embodiments, the cycloalkyl group contains 0, 1, or 2 double bonds. In some embodiments, the cycloalkyl group contains 1 or 2 double bonds (partially unsaturated cycloalkyl). In some embodiments, the cycloalkyl group may be fused with aryl, heteroaryl, cycloalkyl, and heterocyclic alkyl groups. In some embodiments, the cycloalkyl group may be fused with aryl, cycloalkyl, and heterocyclic alkyl groups. In some embodiments, the cycloalkyl group may be fused with aryl and heterocyclic alkyl groups. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cyclohepttrienyl, norcamphenyl, norpinel, norcarel, bicyclo[1.1.1]pentyl, bicyclo[2.1.1]hexyl, and so on.

[0047] Unless otherwise specified, "alkoxy" refers to an alkyl group bonded to the remainder of the molecule via an ether oxygen atom. Representative alkoxy groups are those having 1-6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, and tert-butoxy. As used herein, "alkoxy" includes unsubstituted and substituted alkoxy groups, particularly those substituted with one or more halogens. Preferred alkoxy groups are selected from OCH3, OCF3, CHF2O, CF3CH2O, etc. i- PrO, n- PrO, i- BuO、 n- BuO or t- BuO.

[0048] Unless otherwise specified, "aryl" refers to a hydrocarbon aromatic group, which can be monocyclic or polycyclic, such as a monocyclic aryl ring fused with one or more carbocyclic aromatic groups. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, and phenanthrene.

[0049] Unless otherwise specified, "heteroaryl" refers to an aromatic group containing one or more heteroatoms (O, S, or N), which may be monocyclic or polycyclic. For example, a monocyclic heteroaryl ring may be fused with one or more carbocyclic aromatic groups or other monocyclic heterocyclic alkyl groups. Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyridazinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, quinolinyl, isoquinolinyl, furanyl, thiophene, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrroloyl, indolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzothiaphenyl, benzooxazolyl, benzopyridinyl, pyrrolopyrimidinyl, 1H-pyrrolo[3,2-b]pyridinyl, 1H-pyrrolo[2,3-c]pyridinyl, 1H-pyrrolo[3,2-c]pyridinyl, 1H-pyrrolo[2,3-b]pyridinyl,

[0050] Unless otherwise specified, "halogen" (or halogenated group) means fluorine, chlorine, bromine or iodine. The term "halogenated" (or "halogen substituted") appearing before the group name indicates that the group is partially or completely halogenated, that is, substituted by F, Cl, Br or I in any combination, preferably substituted by F or Cl.

[0051] "Optional" or "optionally" means that the event or condition described below may, but is not required, occur, and the description includes both the scenario where said event or condition occurs and the scenario where said event or condition does not occur.

[0052] The term "membered ring" includes any ring structure. The term "membered" refers to the number of skeleton atoms that make up the ring. For example, cyclohexyl, pyridyl, pyranyl, and thioranyl are six-membered rings, while cyclopentyl, pyrroleyl, furanyl, and thiophenyl are five-membered rings.

[0053] The term "fragment" refers to a specific part or functional group of a molecule. Chemical fragments are generally considered to be chemical entities contained in or attached to a molecule.

[0054] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key The relative configuration of the center of a solid is indicated by a wavy line. Indicates wedge-shaped solid line key or wedge-shaped dashed key Or use wavy lines Indicates a straight solid line key Or straight dashed key

[0055] Unless otherwise stated, use Indicates a single bond or a double bond.

[0056] Specific pharmaceutical and medical terminology

[0057] The term “acceptable,” as used herein, means that a prescription component or active ingredient does not have an excessively harmful effect on the health of a therapeutic target.

[0058] The terms “treatment,” “treatment process,” or “therapy” as used herein include alleviating, suppressing, or improving symptoms or conditions of a disease; suppressing the development of complications; improving or preventing underlying metabolic syndromes; suppressing the development of a disease or symptom, such as controlling the progression of a disease or condition; reducing a disease or symptom; alleviating a disease or symptom; reducing complications arising from a disease or symptom; or preventing or treating signs arising from a disease or symptom. As used herein, a compound or pharmaceutical composition, when administered, may improve a disease, symptom, or condition, particularly by improving its severity, delaying its onset, slowing its progression, or reducing its duration. Whether administered regularly or intermittently, continuously or intermittently, it may be attributable to or related to the administration.

[0059] "Active ingredient" refers to the compound represented by general formula (1), and pharmaceutically acceptable inorganic or organic salts of compounds of general formula (1). The compounds of the present invention may contain one or more asymmetric centers (chiral centers or axial chirality), and thus appear as racemates, racemic mixtures, single enantiomers, diastereomers, and single diastereomers. The asymmetric centers that may exist depend on the nature of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers, and all possible optical isomers and diastereomer mixtures, as well as pure or partially pure compounds, are included within the scope of the present invention. The present invention means including all such isomeric forms of these compounds.

[0060] The terms “compound,” “composition,” “agent,” or “medicine or medicament” may be used interchangeably here, and all refer to a compound or composition that, when applied to an individual (human or animal), can induce a desired pharmaceutical and / or physiological response through local and / or systemic action.

[0061] The term “administered, administering, or administration” here refers to the direct application of the compound or composition described herein, or the application of a prodrug, derivative, or analog of the active compound.

[0062] While the numerical ranges and parameters used to define the broader scope of this invention are approximate values, the relevant values ​​in the specific embodiments have been presented as precisely as possible. However, any value inevitably contains standard deviations due to individual test methods. Here, "approximately" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a particular value or range. Alternatively, the term "approximately" may mean that the actual value falls within the acceptable standard error of the average, as determined by those skilled in the art. Except for experimental examples, or unless expressly stated otherwise, it is understood that all ranges, quantities, values, and percentages used herein (e.g., to describe material usage, duration, temperature, operating conditions, quantity ratios, and others similar) are modified with "approximately". Therefore, unless otherwise stated, the numerical parameters disclosed in this specification and the accompanying claims are approximate values ​​and are subject to change as needed. At a minimum, these numerical parameters should be understood as the indicated significant digits and values ​​obtained using a rounding method.

[0063] Unless otherwise defined in this specification, scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art. Furthermore, unless conflicting with the context, singular nouns used herein include their plural forms, and vice versa.

[0064] Therapeutic uses

[0065] The present invention provides methods for treating diseases using compounds or pharmaceutical compositions of general formula (1) of the present invention, including but not limited to conditions involving DHODH (e.g., cancer).

[0066] In some embodiments, a method for treating cancer is provided, the method comprising administering to an individual in need an effective amount of any of the aforementioned pharmaceutical compositions comprising a compound of general structural formula (1). In other embodiments, the cancer is a hematologic malignancy and a solid tumor, including but not limited to leukemia, breast cancer, lung cancer, pancreatic cancer, colon cancer, bladder cancer, brain cancer, urothelial carcinoma, prostate cancer, liver cancer, ovarian cancer, head and neck cancer, gastric cancer, mesothelioma, or all metastatic cancers.

[0067] route of administration

[0068] The compounds of this invention and their pharmaceutically acceptable salts can be formulated into various preparations, comprising, within a safe and effective range, the compounds of this invention or their pharmaceutically acceptable salts and pharmacologically acceptable excipients or carriers. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. The safe and effective range of the compound is determined based on the age, condition, and duration of treatment of the patient.

[0069] "Pharmaceutically acceptable excipients or carriers" refers to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with and with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmacologically acceptable excipients or carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), and emulsifiers (such as Tween). Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0070] When applying the compounds of this invention, they can be administered orally, rectally, parenterally (intravenously, intramuscularly, or subcutaneously), or topically.

[0071] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following components: (a) fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

[0072] Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

[0073] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0074] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0075] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0076] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0077] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0078] The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable compounds. When using the pharmaceutical composition, a safe and effective amount of the compound of this invention is applied to the mammal (such as a human) requiring treatment, wherein the dose administered is a pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 50–1000 mg. Of course, the specific dosage should also consider factors such as the route of administration and the patient's health condition, which are all within the scope of a skilled physician's expertise.

[0079] The features mentioned above in this invention, or the features mentioned in the embodiments, can be combined arbitrarily. All features disclosed in this specification can be used with any compositional form, and each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar pharmaceutical composition. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features. Detailed Implementation

[0080] The following description will elaborate on the specific aspects, characteristics, and advantages of the aforementioned compounds, methods, and pharmaceutical compositions, making the content of this invention readily apparent. It should be understood that the detailed descriptions and examples described below are specific embodiments and are for reference only. After reading this description, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by this application.

[0081] In all embodiments, 1 H-NMR was recorded using a Varian Mercury 400 NMR spectrometer, and chemical shifts are expressed as δ (ppm). Unless otherwise specified, the silica gel used for separation was 200-300 mesh, and all eluent ratios were by volume.

[0082] The following abbreviations are used in this invention: AlMe3 represents trimethylaluminum; Ar represents argon; CDCl3 represents deuterated chloroform; (COCl)2 represents oxaloyl chloride; Cs2CO3 represents cesium carbonate; DCM represents dichloromethane; Dioxane represents 1,4-dioxane; DMF represents N,N-dimethylformamide; DMSO represents dimethyl sulfoxide; EA or EtOAc represents ethyl acetate; h represents hour; IPA represents isopropanol; KF represents potassium fluoride; KHMDS represents potassium bis(trimethylsilyl)aminobis(trimethyl ... LC-MS stands for Liquid Chromatography-Mass Spectrometry; Me3OBF4 represents Trimethyloxonium tetrafluoroborate; MeOH represents Methanol; mL represents Milliliter; min represents Minute; MS represents Mass Spectrometry; NaH represents Sodium Hydride; Na2SO4 represents Sodium Sulfate; NMR represents Nuclear Magnetic Resonance; Pd2(dba)3 represents Tris(dibenzylacetone)dipalladium; PE represents Petroleum Ether; py represents Pyridine; Xantphos represents 4,5-Bisdiphenylphosphine-9,9-Dimethyloxanthracene; THF represents Tetrahydrofuran; Tol represents Toluene.

[0083] Example 1: Synthesis of (S)-N-(2-chloro-6-fluorophenyl)-5-fluoro-4-(3'-oxo-5',6'-dihydro-3'H-spiro[cyclopropane-1,7-[1,2,4]triazolo[4,3-a]pyridine]-2'(8'H)-yl)-2-((1,1,1-trifluoroprop-2-yl)oxy)benzamide (compound 1)

[0084]

[0085] Synthesis route:

[0086]

[0087] Step 1: Synthesis of Compounds 1-2

[0088] Add 1-1 (4.6 g, 36.75 mmol) and DCM (100 mL) to a 250 mL single-necked flask. After Ar displacement protection, add Me3OBF4 (8.2 g, 55.12 mmol). Stir the mixture at room temperature for 20 h under Ar protection. Then add Me3OBF4 (2.73 g, 28.46 mmol) and continue stirring at room temperature for another 20 h. LC-MS showed that the reaction was basically complete. Quench the mixture with ice water (50 mL), adjust the pH to 7-8 with saturated sodium bicarbonate, separate the phases, extract the aqueous phase with DCM (50 mL), combine the organic phases, wash with saturated sodium chloride solution (50 mL), dry with anhydrous Na2SO4, filter, and concentrate to obtain a yellow-brown solid crude product 1-2 (3.6 g, yield 70.4%). ESI-MS m / z: 140.1 [M+H] + .

[0089] Step 2: Synthesis of compounds 1-3

[0090] In a 250 mL single-necked flask, 1-2 (3.6 g, 25.86 mmol) and toluene (80 mL) were added. After Ar displacement protection, ethyl hydrazine carboxylate (4.04 g, 38.79 mmol) was added. The mixture was stirred at 120 °C for 40 h under Ar protection. After the reaction was completed, the mixture was concentrated to dryness. The residue was purified by column chromatography (DCM:MeOH = 100:0 to 100:1 to 60:1 to 20:1) to give a white solid product 1-3 (1.77 g, yield 41.4%), ESI-MS m / z: 166.1 [M+H]. + .

[0091] Step 3: Synthesis of compounds 1-5

[0092] (S)-1,1,1-trifluoroprop-2-ol (2.726 g, 23.90 mmol) and dry THF (30 mL) were added to a 100 mL three-necked flask. The mixture was cooled to -5 °C to -10 °C in an ice-salt bath under Ar protection. NaH (860 mg, 60%, 21.51 mmol) was added in portions. After the addition was complete, the mixture was kept warm and stirred for 1 h. Then 1-4 (3.0 g, 11.95 mmol) was added, the ice bath was removed, and the reaction was stirred at room temperature for about 3 h. After the reaction was completed, EA (20 mL) was added to the mixture, followed by quenching with ice water (20 mL). The mixture was stirred, separated, and the aqueous phase was extracted again with EA (20 mL). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered, and concentrated to obtain a colorless, transparent oily product 1-5 (2.99 g, yield 72.5%). ESI-MS m / z: 345.0 / 347.0 [M+H] + .

[0093] Step 4: Synthesis of compounds 1-6

[0094] 2-Chloro-6-fluoroaniline (6.11 g, 42.0 mmol) and dry DCM (60 mL) were added to a 250 mL three-necked flask. After Ar displacement protection, trimethylaluminum (42 mL, 1 M in n-Heptane, 42 mmol) was added dropwise at room temperature. After the addition was complete, the mixture was stirred at room temperature for 2 h. Then 1-5 (3.62 g, 10.5 mmol) was added, and the mixture was Ar displacement. The temperature was then raised to 50 °C and stirred for 3 h. After the reaction was completed, the reaction solution was slowly poured into a saturated ammonium chloride solution (100 mL) under ice bath conditions. The mixture was stirred in an ice bath for 30 min, then separated. The aqueous phase was extracted three times with DCM (100 mL * 3). The combined organic phases were washed with a saturated sodium chloride solution (100 mL), concentrated, and the residue purified by column chromatography (PE:EA = 40:0 to 40:1 to 20:1 to 10:1) to give a yellowish-brown gelatinous solid product 1-6 (4.46 g, yield 90.1%). ESI-MS m / z: 458.0 [M+H] + .

[0095] Step 5: Synthesis of Compound 1

[0096] Add 1-6 (500 mg, 1.09 mmol) and 1-3 (234 mg, 1.42 mmol), Cs₂CO₃ (710 mg, 2.18 mmol), Xantphos (378 mg, 0.654 mmol), and Dioxane (10 mL) to a 30 mL microwave reactor. After Ar replacement, add Pd₂(dba)₃ (300 mg, 0.327 mmol). Incubate at 110 °C with stirring for 15 h under microwave. After the reaction is complete as detected by LC-MS, filter the mixture, wash the filter cake with EA (approximately 10 mL), concentrate the filtrate, and purify the residue by column chromatography (PE:EA = 4:0 to 4:1 to 3:1 to 2:1) to obtain approximately 400 mg of impurities. Further purification by prep-HPLC yields a white, foamy solid product, compound 1 (340 mg, yield 57.5%, HPLC > 99%).

[0097] 1H NMR (400MHz, CDCl3) δ: 9.01 (s, 1H), 8.17 (d, J=11.6Hz, 1H), 7.51 (d, J=5.6Hz, 1H), 7.28 (dt, J=8.4, 1.3Hz, 1H), 7.25-.20 (m, 1H), 7.12 (ddd, J=9 .6, 8.2, 1.6Hz, 1H), 4.94 (p, J=6.2Hz, 1H), 3.78 (t, J=6.2Hz, 2H), 2.67 (s, 2H), 1.82 (t, J=6.1Hz, 2H), 1.65 (d, J=6.8Hz, 3H), 0.61 (s, 4H), ESI-MS m / z: 543.2[M+H] + .

[0098] Examples 2-53: Synthesis of Compounds 2-53

[0099] Using different raw materials, target compounds 2-53 were obtained by a similar synthesis method as described in Example 1.

[0100] Table 1

[0101]

[0102]

[0103]

[0104]

[0105] Example 54 Synthesis of (S)-N-(2-chloro-6-fluorophenyl)-5-fluoro-4-(3'-oxo-5',6'-dihydro-3'H-spiro[cyclopropane-1,7-[1,2,4]triazolo[4,3-a]pyridine]-2'(8'H)-yl)-2-((1,1,1-trifluoroprop-2-yl)oxy)benzamide (compound 54)

[0106]

[0107] Synthesis route:

[0108]

[0109] Step 1: Synthesis of Compound 54-2

[0110] Add 10 g (47.62 mmol) of 54-1, 50 mL of THF, and 0.5 mL of DMF to a 250 mL single-necked flask. Cool the mixture to 0–5 °C in an ice bath under argon protection, then add 6.35 g (50 mmol) of (COCl)2 dropwise. After the addition is complete, allow the mixture to rise naturally to room temperature and stir for 2 hours. After the reaction is essentially complete as detected by LC-MS, concentrate the mixture to dryness. Dissolve the resulting residue in 50 mL of THF for later use.

[0111] In a separate 250 mL single-necked flask, add THF (40 mL), isopropanol (4.3 g, 71.43 mmol), and pyridine (4.5 g, 57.14 mmol). Cool the mixture to 0–5 °C in an ice bath under argon protection, then add the THF solution from the previous reaction. After the addition is complete, allow the mixture to naturally rise to room temperature and stir for 1 h. After the reaction is complete as detected by LC-MS, add EA (50 mL) and ice water (50 mL), stir, separate the layers, and extract the aqueous phase again with EA (50 mL). Combine the organic phases, wash with saturated sodium chloride solution (50 mL), concentrate under reduced pressure, and purify the residue by column chromatography (PE:EA = 30:0 to 30:1) to obtain a colorless liquid product 54-2 (11.31 g, yield: 94%), ESI-MS m / z: 252.0 [M+H]. + .

[0112] Step 2: Synthesis of Compound 54-3

[0113] 54-2 (600 mg, 2.38 mmol), compound 1-3 (393 mg, 2.38 mmol), K2CO3 (493 mg, 3.57 mmol), and DMSO (10 mL) were added to a 100 mL single-necked flask. The mixture was heated to 80 °C and stirred for 3 h under argon protection. After the reaction was completed, EA (30 mL) and water (50 mL) were added to the mixture. The mixture was stirred, separated, and the aqueous phase was extracted again with EA (20 mL). The combined organic phases were washed with saturated sodium chloride solution (20 mL), concentrated under reduced pressure, and the residue was purified by column chromatography (PE∶EA = 30∶0 to 3∶1 to 2∶1) to give a yellow gelatinous product 54-3 (820 mg, yield 90%). ESI-MS m / z: 381.1 [M+H] + .

[0114] Step 3: Synthesis of compound 54-4

[0115] 54-3 (820 mg, 2.153 mmol), (S)-1,1,1-trifluoropropane-2-ol (737 mg, 6.46 mmol), KF (375 mg, 6.46 mmol), and DMSO (10 mL) were added to a 100 mL single-necked flask. The mixture was heated to 100 °C under argon protection and stirred for 20 h. LC-MS analysis indicated the reaction was incomplete, so (S)-1,1,1-trifluoropropane-2-ol (368 mg, 3.23 mmol) was added, and the mixture was stirred for another 20 h. Then, (S)-1,1,1-trifluoropropane-2-ol (368 mg, 3.23 mmol) was added again, and the mixture was stirred for another 40 h. LC-MS analysis showed the reaction was essentially complete. The mixture was added to EA (30 mL) and water (30 mL), stirred, and separated. The aqueous phase was extracted again with EA (20 mL). The combined organic phases were washed with saturated sodium chloride solution (20 mL), concentrated under reduced pressure, and the residue was purified by column chromatography (PE∶EA = 30∶0 to 3∶1 to 2∶1) to give a light brown foamy solid product 54-4 (543 mg, yield 55%). ESI-MS m / z: 459.1 [M+H] + .

[0116] Step 4: Synthesis of Compound 54

[0117] 2-Chloro-6-fluoroaniline (685 mg, 4.712 mmol) was added to a 250 mL three-necked flask. After drying with 10 mL of DCM and purging with argon, trimethylaluminum (47 mL, 1 M in n-Heptane, 4.7 mmol) was added dropwise at room temperature. After the addition was complete, the mixture was stirred at room temperature for 2 h. Then, 54-4 (540 mg, 1.178 mmol) was added, and the mixture was purged with argon before being heated to 50 °C and stirred for 3 h. After the reaction was completed, the reaction solution was slowly poured into a saturated ammonium chloride solution (20 mL) under ice bath conditions. The mixture was stirred in an ice bath for 30 min, then separated. The aqueous phase was extracted again with DCM (20 mL * 3). The combined organic phases were washed with a saturated sodium chloride solution (20 mL), concentrated, and the residue was purified by column chromatography (PE:EA = 3:0 to 3:1 to 2:1) to obtain a white, foamy solid product (455 mg, yield: 71.1%). ESI-MS m / z: 544.0 [M+H] + .

[0118] 1H NMR (400MHz, Chloroform-d) δ9.10 (s, 1H), 8.54 (d, J = 9.4Hz, 1H), 7.27 (d, J = 7.3Hz, 2H), 7.14 (d, J = 8.9Hz, 1H ), 5.97 (s, 1H), 3.78 (t, J = 6.1Hz, 2H), 2.68 (s, 2H), 1.82 (t, J = 6.1Hz, 2H), 1.67 (d, J = 6.5Hz, 3H), 0.62 (s, 4H).

[0119] Examples 55-59: Synthesis of Compounds 55-59

[0120] Using different raw materials, target compounds 55-59 were obtained by a synthesis method similar to that in Example 54.

[0121] Table 2

[0122]

[0123]

[0124] The control compound used in this invention is BAY-2402234, and its synthesis method is based on patent WO2018077923. Its structure is shown below:

[0125]

[0126] Example 60: Determination of the DHODH enzyme activity by the compounds of the present invention.

[0127] The activity of DHODH enzyme was determined by measuring its ability to catalyze the oxidation of the natural substrate DHO (dihydroorotic acid) to orotic acid via a cascade reaction, while simultaneously reducing coenzyme Q. The activity of DHODH was indirectly detected by measuring the ability of coenzyme Q to reduce the chromogenic substrate DCIP. After incubating the compound and enzyme reaction mixture at a warm temperature for 5 minutes, the reaction substrate DHO was added to initiate the reaction. Enzyme activity was determined by measuring the depletion of DCIP at 600 nm at regular intervals. Compared with the control group, the inhibition rate of DHODH enzyme activity at different concentrations of the compound was calculated, and the half-maximal inhibitory concentration (IC50) was calculated. 50 The values ​​and filtering results are shown in Table 3.

[0128] Example 61: Determination of the antiproliferative activity of the compounds of the present invention against THP-1 cells.

[0129] THP-1 cells were seeded in 384-well plates (Fisher 142762) at 3000 cells per well. On the second day, serially diluted compounds were added. 72 hours after compound addition, CellTiter-Lumi (Beyotime C0068XL) was added to measure ATP levels in the cells, evaluating cell growth and calculating the IC50 of the compound's inhibitory effect on cell growth. 50 The screening results are shown in Table 3.

[0130] Example 62: Determination of the antiproliferative activity of the compounds of the present invention against MV-4-11 cells.

[0131] MV-4-11 cells were seeded in 384-well plates (Fisher 142762) at 3000 cells per well. On the second day, serially diluted compounds were added. 72 hours after compound addition, CellTiter-Lumi (Beyotime C0068XL) was added to measure ATP levels in the cells, evaluating cell growth and calculating the IC50 of the compound's inhibitory effect on cell growth. 50 The screening results are shown in Table 3.

[0132] Table 3. Inhibitory activity of the compounds of the present invention against DHODH enzyme, and antiproliferative activity against THP-1 and MV-4-11 cells.

[0133]

[0134]

[0135] A represents IC 50 ≤50nM

[0136] B represents 50nM≤IC 50 ≤0.5μM

[0137] C represents IC 50 >0.5μM

[0138] As shown in the table above, the compounds of this invention exhibit strong anti-proliferative activity against DHODH enzyme and against tumor cells THP-1 and MV-4-11. Furthermore, compared to the control compound, the compounds of this invention show a significant increase in inhibitory activity against MV-4-11 cells.

[0139] Example 63: Pharmacokinetic Evaluation in Mice

[0140] The compound was administered via intravenous injection and oral gavage. The intravenous dose was 2 mg / kg, and the oral dose was 10 mg / kg (0.5% CMC-Na suspension). Fifteen male ICR mice were used, and three mice were sampled at three discontinuous time points per mouse. Sampling time points were before administration, and 5 min, 15 min, 30 min, 1 h, 3 h, 5 h, 8 h, 12 h, and 24 h after administration. Approximately 80 μL of blood was collected from the orbital cavity or heart at each time point after administration. All whole blood samples were collected in EDTAK2-containing tubes, centrifuged at 4 °C (1500-1600 rpm / min) for 10 min, and the plasma was separated and stored at -90 to -60 °C for analysis. The concentration of the compound in plasma was determined by liquid chromatography-tandem mass spectrometry, and the corresponding pharmacokinetic parameters were calculated based on the plasma concentration-time curve.

[0141] Table 4. Pharmacokinetic parameters of compound 1 in mice.

[0142]

[0143] NA indicates that the data is unavailable.

[0144] The data in the table above show that compound 1 has good oral bioavailability and good oral absorption properties, which is of great significance in improving drug efficacy, reducing dosage, and saving costs.

[0145] While specific embodiments of the present invention have been described above, those skilled in the art should understand that these are merely illustrative examples, and various changes or modifications can be made to these embodiments without departing from the principles and essence of the present invention. Therefore, the scope of protection of the present invention is defined by the appended claims.

Claims

1. A compound of formula (1) or a pharmaceutically acceptable salt thereof: ； In general formula (1): n is 0, 1, or 2; X is CH or N; R 1 It is phenyl, pyridyl or pyrazolyl, and the phenyl, pyridyl or pyrazolyl may be substituted by one or more of the following groups: halogen, NH2, C1-C3 alkyl, C1-C3 alkoxy or halo-C1-C3 alkyl; R 2 For H or F; R 3 It is a C1-C3 alkyl group; R 4 It is a C1-C6 alkyl, C3-C8 cycloalkyl, or halogenated C1-C6 alkyl.

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein in the general formula (1), R 1 for: , , , or , where R a R b R c R d and R e It can be H, F, Cl, Me, Et, OMe, CHF2, CF3 or NH2 independently.

3. The compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein in the general formula (1), R 3 For Me.

4. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein in the general formula (1), R 4 For Me, Et, CF3, , , , , or .

5. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has one of the following structures: 。 6. A pharmaceutical composition, characterized in that, It contains a pharmaceutically acceptable excipient or carrier, and a compound or a pharmaceutically acceptable salt thereof as any one of claims 1-5 as the active ingredient.

7. The use of a compound as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 6, in the preparation of a medicament for the prevention or treatment of tumors.

8. The application as described in claim 7, wherein the tumor includes solid tumors and hematomas.

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