1,2,4-Triazolone derivatives as DHODH inhibitors

Abstract

The present invention relates to 1,2,4-triazolone derivatives used as DHODH inhibitors. Specifically, the present invention relates to compounds represented by general formula (1) and a preparation method thereof, and use of the compounds represented by general formula (1) and their optical isomers, crystal forms, and pharmaceutically acceptable salts as DHODH inhibitors in the treatment of DHODH-related diseases.

Description

[0001] This application claims the priority of Chinese application CN202011042241.4, filed on September 28, 2020. This application cites the entire text of the above Chinese application. Technical Field

[0002] The present invention relates to the field of pharmaceutical chemistry, and more specifically, to a novel 1,2,4-triazolone derivative, a preparation method thereof and the use of the compound. Background Art

[0003] Dihydroorotate Dehydrogenase (DHODH) is an iron-containing flavin-dependent enzyme that is mainly present on the inner membrane of mitochondria. It is a key enzyme for the de novo synthesis of pyrimidine nucleotides. Pyrimidine nucleotides are the backbone components of DNA and RNA. Orotic acid is a precursor for the synthesis of pyrimidine nucleotides. DHODH catalyzes the dehydrogenation of dihydroorotate, converting it into orotic acid. Therefore, inhibiting DHODH can block the de novo synthesis of pyrimidine, resulting in DNA and RNA synthesis disorders.

[0004] Pyrimidine nucleotides in cells mainly come from de novo synthesis and salvage synthesis pathways. For normal differentiated resting cells, pyrimidine nucleotides mainly come from the salvage, also known as the regeneration synthesis pathway, which directly converts free pyrimidine bases into pyrimidine nucleotides. However, for T and B lymphocytes with vigorous metabolism or rapidly dividing cancer cells under pathological conditions, the pyrimidine nucleotides generated by the salvage pathway cannot maintain the function or survival of the cells. Therefore, relying on DHODH to synthesize pyrimidine nucleotides from scratch is crucial for the survival of such cells. Inhibiting the activity of DHODH can selectively prevent the large demand for pyrimidine nucleotides in such cells, thereby hindering the synthesis of biological macromolecules such as DNA, RNA, and glycoproteins, and inhibiting cell function or growth.

[0005] Taking advantage of this property, the development of DHODH inhibitors targeting abnormally activated T and B cells and rapidly dividing cancer cells for the purpose of treating cancer, viral infections, autoimmune diseases (rheumatoid arthritis and multiple sclerosis) has achieved initial results. For example, the DHODH small molecule inhibitors leflunomide / teriflunomide and buquina have been approved for the treatment of rheumatoid arthritis and multiple sclerosis. However, these inhibitors are highly lipid-soluble and poorly selective, and long-term use will produce significant toxic side effects. Therefore, finding new, efficient, and low-side-effect DHODH inhibitors for the treatment of immune-related diseases and tumors has become a hot topic in current research. Summary of the invention

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

[0007]

[0008] In the general formula (1):

[0009] n is 0, 1 or 2;

[0010] X is -CH2-, -O-, -S-, -S(O)-, -S(O)2- or -N(R 5 )-;

[0011] R 1 is an aryl or heteroaryl group, and the aryl and heteroaryl groups may be substituted by one or more of the following groups: halogen, OH, NH2, CN, C1-C3 alkyl, C1-C3 alkoxy, C3-C6 cycloalkyl, halogenated C1-C3 alkyl or halogenated C1-C3 alkoxy;

[0012] R 2 is H or F;

[0013] R 3 is C1-C3 alkyl or C3-C8 cycloalkyl;

[0014] R 4 C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl or C3-C6 cycloalkyl substituted C1-C3 alkyl;

[0015] R 5 It is H, C1-C3 alkyl or C3-C6 cycloalkyl.

[0016] In another preferred embodiment, the isomers are optical isomers.

[0017] In another preferred embodiment, in the general formula (1), X is -CH2-, -O-, -S-, -S(O)-, -S(O)2-, -NH-, -N(Me)- or -N(Et)-; preferably X is -CH2-, -O-, -S-, -S(O)-, -S(O)2- or -N(Me)-, more preferably X is -CH2- or -O-, and more preferably X is -CH2-.

[0018] In another preferred embodiment, in the general formula (1), R 1 for: Where R a , R b , R c , R d and R eare independently H, F, Cl, Me, Et, OMe, OEt, CHF2, CF3, OH, NH2 or CN.

[0019] In another preferred embodiment, in the general formula (1), R 3 For me.

[0020] In another preferred embodiment, in the general formula (1), R 4 Me, Et, CF3, Preferred R 4 It is CF3.

[0021] In another specific embodiment of the present invention, the compound of formula (1) has one of the following structures:

[0022]

[0023]

[0024]

[0025]

[0026]

[0027] The present invention also provides a pharmaceutical composition, which contains a pharmaceutically acceptable carrier, diluent and / or excipient, and a compound of the general formula (1) of the present invention, or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates as active ingredients.

[0028] The present invention also provides the use of the compound represented by the general formula (1) of the present invention, or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates, or the above-mentioned pharmaceutical compositions for preparing drugs for treating, regulating or preventing DHODH-related diseases.

[0029] The present invention also provides a method for treating, regulating or preventing DHODH-related diseases, comprising administering to a subject a therapeutically effective amount of a compound represented by the general formula (1) of the present invention, or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates, or the above-mentioned pharmaceutical composition.

[0030] In another specific embodiment, the DHODH-related disease is a DHODH-mediated related disease.

[0031] In another specific embodiment, the DHODH-related disease is cancer.

[0032] In another specific embodiment, the cancer is a solid tumor or a hematological tumor.

[0033] In another specific embodiment, the cancer is leukemia, breast cancer, lung cancer, pancreatic cancer, colon cancer, bladder cancer, brain cancer, urothelial cancer, prostate cancer, liver cancer, ovarian cancer, head and neck cancer, gastric cancer, mesothelioma or all cancer metastases.

[0034] It is to be understood that both the foregoing general description and the following detailed description of the present invention are exemplary and explanatory and are intended to provide further explanation of the invention as claimed.

[0035] Synthesis of compounds

[0036] The following is a detailed description of the preparation methods of the compounds of the general formula (1) of the present invention, but these specific methods do not constitute any limitation to the present invention.

[0037] The compounds of formula (1) described above can be synthesized using standard synthetic techniques or known techniques in combination with the methods described herein. In addition, the solvents, temperatures and other reaction conditions mentioned herein can be varied. The starting materials used in the synthesis of the compounds can be synthesized or obtained from commercial sources. The compounds described herein and other related compounds having different substituents can be synthesized using known techniques and starting materials, including those found in March, ADVANCED ORGANIC CHEMISTRY 4 th Ed., (Wiley 1992); Carey and Sundberg, ADVANCED ORGANIC CHEMISTRY 4 th Ed., Vols.A and B (Plenum 2000, 2001), Green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS3 rd Ed., (Wiley 1999). The general methods for the preparation of compounds can be modified by using appropriate reagents and conditions to introduce various groups into the formulae provided herein.

[0038] 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, the amount of the compounds used, the reaction temperature, the time required for the reaction, etc. are not limited to the following explanation. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such a combination can be easily performed by a person skilled in the art to which the present invention belongs. On the one hand, the present invention also provides a method for preparing the compound represented by the general formula (1), wherein the compound of the general formula (1) can be prepared by the following method A:

[0039] Method A

[0040] Method A comprises the following steps: firstly, the carboxyl group of compound A1 is subjected to chlorination reaction to generate acyl chloride compound A2; compound A2 is further reacted with isopropanol under the action of a strong base to generate compound A3; compound A3 and compound A4 are reacted under alkaline conditions to generate compound A5; compound A6 is reacted under strong alkaline conditions and then reacted with compound A5 to obtain compound A7; compound A7 and a suitable raw material R are reacted to obtain compound A5; 1 NH2 reaction gave the target compound A8.

[0041]

[0042] In the above reaction equation, n, X, R 1 , R 2 , R 3 and R 4 The definition of is as mentioned above.

[0043] Further forms of compounds

[0044] "Pharmaceutically acceptable" as used herein refers to a material, such as a carrier or diluent, that does not abrogate the biological activity or properties of the compound and is relatively non-toxic, i.e., a material that, when administered to a subject, does not cause undesirable biological effects or interact in a deleterious manner with any of its constituent components.

[0045] The term "pharmaceutically acceptable salt" refers to a form of a compound that does not cause significant irritation to the organism to which it is administered and does not cause the biological activity and properties of the compound to disappear. In certain specific aspects, the pharmaceutically acceptable salt is obtained by reacting the compound of formula (1) with an acid, such as an inorganic acid such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, phosphoric acid, 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, p-toluenesulfonic acid, and an acidic amino acid such as aspartic acid and glutamic acid.

[0046] It should be understood that references to pharmaceutically acceptable salts include solvent-added forms or crystal forms, particularly solvates or polymorphs. Solvates contain stoichiometric or non-stoichiometric amounts of solvent and are selectively formed during crystallization with pharmaceutically acceptable solvents such as water, ethanol, etc. Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is ethanol. Solvates of compounds of formula (1) are conveniently prepared or formed according to the methods described herein. For example, hydrates of compounds of formula (1) are conveniently prepared by recrystallization from a mixed solvent of water / organic solvent, and the organic solvents used include, but are not limited to, tetrahydrofuran, acetone, ethanol or methanol. In addition, the compounds mentioned herein can exist in unsolvated and solvated forms. In general, for the purposes of the compounds and methods provided herein, the solvated form is considered to be equivalent to the unsolvated form.

[0047] In other specific embodiments, the compound of formula (1) is prepared in different forms, including but not limited to amorphous, crushed and nano-particle forms. In addition, the compound of formula (1) includes crystalline forms and can also be used as polymorphs. Polymorphs include different lattice arrangements of the same elemental composition of the compound. Polymorphs usually have different X-ray diffraction spectra, infrared spectra, melting points, density, hardness, crystal form, optical and electrical properties, stability and solubility. Different factors such as recrystallization solvents, crystallization rate and storage temperature may cause a single crystal form to dominate.

[0048] In another aspect, the compounds of formula (1) may have chiral centers and / or axial chirality, and therefore appear in the form of racemates, racemic mixtures, single enantiomers, diastereomeric compounds 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 diastereomeric mixtures as well as pure or partially pure compounds are included within the scope of the present invention. The present invention is meant to include all such isomeric forms of these compounds.

[0049] The compounds of the invention may contain unnatural proportions of atomic isotopes on one or more of the atoms that make up 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 another example, deuterium can be used to replace hydrogen atoms to form deuterated compounds. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs generally have the advantages of reducing toxic side effects, increasing drug stability, enhancing therapeutic effects, and extending the half-life of drugs in vivo. All isotopic composition changes of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.

[0050] the term

[0051] Unless otherwise indicated, the terms used in this application, including the specification and claims, are defined as follows. It must be noted that in the specification and the appended claims, the singular forms "a" and "an" include plural meanings unless otherwise clearly indicated in the text. If not otherwise indicated, conventional methods of mass spectrometry, nuclear magnetic resonance, HPLC, protein chemistry, biochemistry, recombinant DNA technology and pharmacology are used. In this application, if not otherwise indicated, the use of "or" or "and" means "and / or".

[0052] Unless otherwise specified, "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight and branched groups of 1 to 6 carbon atoms. Preferred are lower alkyl groups containing 1 to 4 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl. As used herein, "alkyl" includes unsubstituted and substituted alkyl groups, especially alkyl groups substituted by one or more halogens. Preferred alkyl groups are selected from CH3, CH3CH2, CF3, CHF2, CF3CH2, CF3(CH3)CH, i Pr, n Pr, i Bu, n Bu or t Bu.

[0053] Unless otherwise specified, "alkenyl" refers to an unsaturated aliphatic hydrocarbon group containing a carbon-carbon double bond, including straight or branched groups of 1 to 14 carbon atoms. Preferably, the lower alkenyl group contains 1 to 4 carbon atoms, such as vinyl, 1-propenyl, 1-butenyl or 2-methylpropenyl.

[0054] Unless otherwise specified, "alkynyl" refers to an unsaturated aliphatic hydrocarbon group containing a carbon-carbon triple bond, including straight and branched chain groups of 1 to 14 carbon atoms. Preferably, the lower alkynyl group contains 1 to 4 carbon atoms, such as ethynyl, 1-propynyl or 1-butynyl.

[0055] Unless otherwise specified, "cycloalkyl" refers to a non-aromatic hydrocarbon ring system (monocyclic, bicyclic or polycyclic), if the carbocyclic ring contains at least one double bond, then the partially unsaturated cycloalkyl may be referred to as "cycloalkenyl", or if the carbocyclic ring contains at least one triple bond, then the partially unsaturated cycloalkyl may be referred to as "cycloalkynyl". Cycloalkyl may include monocyclic or polycyclic (e.g., having 2, 3 or 4 fused rings) groups and spirocycles. In some embodiments, cycloalkyl is monocyclic. In some embodiments, cycloalkyl is monocyclic or bicyclic. The ring-forming carbon atoms of cycloalkyl may be optionally oxidized to form oxo or sulfide groups. Cycloalkyl also includes cycloalkylene. In some embodiments, cycloalkyl contains 0, 1 or 2 double bonds. In some embodiments, cycloalkyl contains 1 or 2 double bonds (partially unsaturated cycloalkyl). In some embodiments, cycloalkyl may be fused with aryl, heteroaryl, cycloalkyl and heterocycloalkyl. In some embodiments, cycloalkyl may be fused with aryl, cycloalkyl and heterocycloalkyl. In some embodiments, cycloalkyl can be fused with aryl and heterocycloalkyl. In some embodiments, cycloalkyl can be fused with aryl and cycloalkyl. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, norbornyl, norpinenyl, norcarbyl, bicyclo[1.1.1]pentanyl, bicyclo[2.1.1]hexanyl, etc.

[0056] Unless otherwise specified, "alkoxy" refers to an alkyl group bonded to the rest of the molecule through an ether oxygen atom. Representative alkoxy groups are those having 1 to 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, especially alkoxy groups substituted with one or more halogens. Preferred alkoxy groups are selected from OCH3, OCF3, CHF2O, CF3CH2O, i- PrO, n- PrO, i- BuO, n- BuO or t- BuO.

[0057] Unless otherwise specified, "aryl" refers to a hydrocarbon aromatic group, which is monocyclic or polycyclic, for example, a monocyclic aryl ring fused to one or more carbocyclic aromatic groups. Examples of aryl include, but are not limited to, phenyl, naphthyl, and phenanthrenyl.

[0058] Unless otherwise specified, "heteroaryl" refers to an aromatic group containing one or more heteroatoms (O, S or N), and the heteroaryl group is monocyclic or polycyclic. For example, a monocyclic heteroaryl ring is fused with one or more carbocyclic aromatic groups or other monocyclic heterocycloalkyl groups. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyridazinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, quinolyl, isoquinolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzothienyl, benzoxazolyl, 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,

[0059] Unless otherwise specified, "halogen" (or halo) refers to fluorine, chlorine, bromine or iodine. The term "halo" (or "halogen substituted") appearing before the name of a group means that the group is partially or fully halogenated, that is, substituted by F, Cl, Br or I in any combination, preferably substituted by F or Cl.

[0060] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0061] When the number of a linking group is 0, such as -(CH2)0-, it means that the linking group is a single bond.

[0062] When one of the variables is selected from a chemical bond, it means that the two groups it connects are directly connected. For example, when L in XLY represents a chemical bond, it means that the structure is actually XY.

[0063] Unless otherwise specified, the key is a solid wedge-shaped key ( ) and dotted wedge bonds ( ) represents the absolute configuration of a stereocenter, with a straight solid bond ( ) and straight dashed bonds ( ) indicates the relative configuration of the stereocenter, and a wavy line ( ) indicates a wedge-shaped solid bond ( ) or a dotted wedge key ( ), or with a wavy line ( ) represents a straight solid bond ( ) or a straight dashed key ( ).

[0064] Unless otherwise stated, Indicates a single bond or a double bond.

[0065] Specific pharmaceutical and medical terms

[0066] The term "acceptable," as used herein, means that a formulation component or active ingredient has no undue deleterious effect on health and well-being for the general purpose of treatment.

[0067] The terms "treat", "treatment" or "therapy" as used herein include alleviating, inhibiting or improving symptoms or conditions of a disease; inhibiting the occurrence of complications; improving or preventing potential metabolic syndrome; inhibiting the occurrence of a disease or symptom, such as controlling the development of a disease or condition; alleviating a disease or symptom; reducing a disease or symptom; alleviating complications caused by a disease or symptom, or preventing or treating signs caused by a disease or symptom. As used herein, a compound or pharmaceutical composition, after administration, can improve a disease, symptom or condition, especially improve its severity, delay the onset, slow the progression of the disease, or reduce the duration of the disease. Whether fixed or temporary administration, continuous administration or intermittent administration, can be attributed to or related to the administration.

[0068] "Active ingredient" refers to the compound shown in the general formula (1), and the pharmaceutically acceptable inorganic or organic salt of the compound of the general formula (1). The compounds of the present invention may contain one or more asymmetric centers (chiral centers or axial chirality), and therefore appear in the form of racemates, racemic mixtures, single enantiomers, diastereomeric compounds and single diastereomers. The asymmetric center that may exist depends on the properties of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers, and all possible optical isomers and diastereomeric mixtures as well as pure or partially pure compounds are included within the scope of the present invention. The present invention is meant to include all such isomeric forms of these compounds.

[0069] The terms "compound", "composition", "agent" or "medicine or medicament" are used interchangeably herein and refer to a compound or composition that, when administered to a subject (human or animal), is capable of inducing a desired pharmaceutical and / or physiological response through local and / or systemic effects.

[0070] The term "administered," "administering," or "administration" as used herein refers to directly administering the compound or composition, or administering a prodrug, derivative, or analog of the active compound.

[0071] Although the numerical ranges and parameters used to define the broader scope of the present invention are approximate values, the relevant numerical values ​​in the specific embodiments have been presented as accurately as possible. However, any numerical value inherently inevitably contains standard deviations due to individual test methods. Here, "about" generally means that the actual value is within plus or minus 10%, 5%, 1% or 0.5% of a specific value or range. Alternatively, the term "about" means that the actual value falls within the acceptable standard error of the mean value, depending on the consideration of those skilled in the art. Except for the experimental examples, or unless otherwise explicitly stated, it should be understood that all ranges, quantities, values ​​and percentages used herein (for example, to describe the amount of material used, the length of time, temperature, operating conditions, quantitative ratios and the like) are modified by "about". Therefore, unless otherwise stated to the contrary, the numerical parameters disclosed in this specification and the attached claims are all approximate values ​​and can be changed as needed. At least these numerical parameters should be understood as the number of significant digits indicated and the values ​​obtained using the general rounding method.

[0072] Unless otherwise defined in this specification, the meanings of scientific and technical terms used herein are the same as those commonly understood by those skilled in the art. In addition, singular nouns used in this specification include plural forms of the nouns, and plural nouns used also include singular forms of the nouns, unless they conflict with the context.

[0073] Therapeutic Uses

[0074] The present invention provides methods for treating diseases using the compounds of formula (1) or pharmaceutical compositions of the present invention, including but not limited to conditions involving DHODH (eg, cancer).

[0075] In some embodiments, a method for treating cancer is provided, comprising administering to an individual in need thereof an effective amount of any of the aforementioned pharmaceutical compositions comprising a compound of formula (1). In some embodiments, the cancer is associated with the synthesis of DHODH. In other embodiments, the cancer is a blood cancer and a solid tumor, including but not limited to leukemia, breast cancer, lung cancer, pancreatic cancer, colon cancer, bladder cancer, brain cancer, urothelial cancer, prostate cancer, liver cancer, ovarian cancer, head and neck cancer, gastric cancer, mesothelioma, or all cancer metastases.

[0076] Route of administration

[0077] The compounds of the present invention and their pharmaceutically acceptable salts can be prepared into various preparations, which contain the compounds of the present invention or their pharmaceutically acceptable salts within the safe and effective amount range and pharmacologically acceptable excipients or carriers. The "safe and effective amount" means that the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. The safe and effective amount of the compound is determined according to the specific circumstances such as the age, condition, and course of treatment of the subject.

[0078] "Pharmaceutically acceptable excipients or carriers" refers to: one or more compatible solid or liquid fillers or gel substances, which 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 the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some 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, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0079] The compounds of the present invention may be administered orally, rectally, parenterally (intravenously, intramuscularly or subcutaneously), or topically.

[0080] 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 ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0081] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0082] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances, etc.

[0083] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0084] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0085] Compositions for parenteral injection may include 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.

[0086] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0087] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds. When using a pharmaceutical composition, a safe and effective amount of the compounds of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 50 to 1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health status, which are all within the skill range of skilled physicians.

[0088] The above features mentioned in the present invention or the features mentioned in the embodiments can be combined in any way. All the features disclosed in the specification of this case can be used in any combination form, and each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features. DETAILED DESCRIPTION

[0089] In the following description, each specific aspect, characteristic and advantage of the above-mentioned compounds, methods and pharmaceutical compositions will be described in detail to make the content of the present invention become very clear. It should be understood that the following detailed description and examples describe specific embodiments and are for reference only. After reading the description of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent situations also fall within the scope defined by the application.

[0090] In all embodiments, 1 H-NMR was recorded using a Varian Mercury 400 nuclear magnetic resonance instrument, and chemical shifts are expressed in δ (ppm). The silica gel used for separation was 200-300 mesh unless otherwise specified, and the ratios of the eluents were all by volume.

[0091] The present invention adopts the following abbreviations: AlMe3 represents trimethylaluminum; Ar represents argon; CDCl3 represents deuterated chloroform; (COCl)2 represents oxalyl chloride; DCM represents dichloromethane; DMF represents dimethylformamide; DMSO represents dimethyl sulfoxide; EA or EtOAc represents ethyl acetate; h represents hour; IPA represents isopropanol; K2CO3 represents potassium carbonate; KHMDS represents potassium bis(trimethylsilyl)amide; LC-MS represents liquid chromatography-mass spectrometry; mL represents milliliters; min represents minutes; MS represents mass spectrometry; Na2SO4 represents sodium sulfate; NMR represents nuclear magnetic resonance; PE represents petroleum ether; py represents pyridine; THF represents tetrahydrofuran.

[0092] Example 1 Synthesis of (S)-N-(2-chloro-6-fluorophenyl)-5-fluoro-6-(3-oxo-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyridin-2(3H)-yl)-2-(((1,1,1-trifluoropropane-2-yl)oxy)nicotinamide (Compound 1)

[0093]

[0094] Synthesis route:

[0095]

[0096] Step 1: Synthesis of compound 1-1

[0097] 2,6-dichloro-5-fluoronicotinic acid (2.1 g, 10.0 mmol), THF (50 mL) and DMF (73 mg, 1.0 mmol) were added to a 100 mL three-necked flask. After the mixed solution was replaced with Ar for protection, it was cooled to 0-5°C in an ice bath, and a solution of (COCl)2 (1.52 g, 12 mmol) in THF (5 mL) was slowly added dropwise. After the addition was completed, the mixed solution was heated to room temperature and stirred for 2 h. After the reaction was completed by LC-MS, the mixed solution was concentrated under reduced pressure to obtain a crude product 1-1 (2.42 g, yield 100%), which was directly used for the next step without purification.

[0098] Step 2: Synthesis of compound 1-2

[0099] The crude product 1-1 (2.42 g, 10.0 mmol) and THF (50 mL) obtained in the previous step were added to a 100 mL single-mouth bottle, and the mixture was cooled in an ice bath after Ar replacement. A mixed solution of isopropanol (1.2 g, 20 mmol) / pyridine (1.2 g, 15.2 mmol) in THF (10 mL) was added dropwise. After the addition was complete, the mixture was heated to room temperature and stirred for 2 h. After the reaction was completed by LC-MS detection, EA (50 mL) and H2O (100 mL) were added to the system, stirred, separated, and the organic phase was washed with a saturated sodium chloride solution (50 mL), dried over anhydrous Na2SO4, and concentrated. The residue was purified by column chromatography (PE / EA = 10 / 0 to 10 / 1) to obtain a colorless oily product 1-2 (2.14 g, yield 85%), ESI-MS m / z: 252 [M+H] + .

[0100] Step 3: Synthesis of Compound 1-3

[0101] 1-2 (2.10 g, 8.333 mmol), DMSO (40 mL), 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyridine-3(2H)-one (1.27 g, 9.17 mmol) and anhydrous K2CO3 (1.72 g, 12.5 mmol) were added to a 100 mL single-necked bottle. After argon replacement, the temperature was raised to 80 °C and the reaction was stirred for 4 h. After the reaction was completed by LC-MS detection, EA (60 mL) and H2O (60 mL) were added to the system, stirred, separated, the aqueous phase was extracted with EA (30 mL), the combined organic phases were washed with saturated sodium chloride solution (30 mL*2), dried over anhydrous Na2SO4, concentrated, and the residue was purified by column chromatography (PE / EA=1 / 0 to 1 / 1) to obtain an off-white solid product 1-3 (2.12 g, yield 72%), ESI-MS m / z: 355 [M+H] + .

[0102] Step 4: Synthesis of Compound 1-4

[0103] (S)-1,1,1-trifluoropropane-2-ol (228 mg, 2.0 mmol) and dry THF (5 mL) were added to a 100 mL three-necked flask, and the temperature was cooled to -10 to -20°C in an ice-salt bath under Ar protection. KHMDS (2.5 mL, 1 M in THF, 2.5 mmol) was then added dropwise. After the addition was complete, the mixture was kept at -10 to -20°C and stirred for 30 min. Then, a solution of compound 1-3 (355 mg, 1.0 mmol) in THF (5 mL) was added dropwise at below -10°C, and the mixture was stirred and reacted for 5 h. After the reaction was completed by LC / LC-MS detection, EA (20 mL) was added to the mixture, and a saturated ammonium chloride solution (20 mL) was used to quench the reaction. The mixture was stirred and separated. The aqueous phase was extracted with EA (20 mL). The combined organic phases were washed with a saturated sodium chloride solution (20 mL*2), dried over anhydrous Na2SO4, and concentrated. The residue was purified by column chromatography (PE / EA=1 / 10 to 1 / 3) to obtain a brown oily product 1-4 (134 mg, yield: 31%), ESI-MS m / z: 433 [M+H] + .

[0104] Step 5: Synthesis of compound 1

[0105] 2-Chloro-6-fluoroaniline (90 mg, 0.62 mmol) and dry DCM (5 mL) were added to a 50 mL three-necked flask, and trimethylaluminum (0.31 mL, 2 M in toluene, 0.62 mmol) was added dropwise at room temperature under argon protection. After the addition was complete, the mixed solution was stirred at room temperature for 30 min, and then a DCM (2 mL) solution of compound 1-4 (134 mg, 0.31 mmol) was added, and the mixture was heated to 50 ° C and stirred to react overnight. After LC-MS detection showed that the raw material was basically gone, the system was quenched with water (5 mL), separated, and the aqueous phase was extracted with DCM (10 mL) again, the organic phases were combined and concentrated, and the residue was purified by pre-TLC (EA / PE=1 / 1) to obtain a light brown solid product 1 (42 mg, yield 26%).

[0106] 1 H NMR (400MHz, CDCl3) δ: 9.51 (s, 1H), 8.14 (d, J=11.3Hz, 1H), 7.37 (dt, J=8.0, 1.4Hz, 1H), 7.24-7.19 (m, 1H), 7.05 (ddd, J=9.6 , 8.1, 1.7Hz, 1H), 4.93 (h, J=6.2Hz, 1H), 3.42 (m, 2H), 2.08-2.02 (m, 2H), 1.67 (m, 4H), 1.56 (d, J=6.5Hz, 3H), LC-MS: 518[M+H] + .

[0107] Example 2 Synthesis of (S)-N-(2-chloro-6-fluorophenyl)-5-fluoro-6-(3-oxo-5,6-dihydro-3H-[1,2,4]triazolo[3,4-c][1,4]oxazin-2(8H)-yl)-2-((1,1,1-trifluoropropan-2-yl)oxy)nicotinamide (Compound 67)

[0108]

[0109] Synthesis route:

[0110]

[0111] Using 1-2 and 2,5,6,8-tetrahydro-3H-[1,2,4]triazolo[3,4-c][1,4]oxazine-3-one as raw materials, a similar synthesis method as in Example 1 was used to obtain the target compound 67.

[0112] 1 H NMR (400MHz, Chloroform-d) δ9.04 (s, 1H), 8.49 (d, J=9.6Hz, 1H), 7.24-7.16 (m, 2H), 7.11-7.03 (m, 1H), 5. 87 (p, J=6.4Hz, 1H), 4.70 (s, 2H), 4.02 (dd, J=6.1, 4.6Hz, 2H), 3.75 (t, J=5.3Hz, 2H), 1.61 (d, J=6.5Hz, 3H).

[0113] Example 3-76 Synthesis of Compounds 2-66, 68-76

[0114] Using synthesis method A and adopting different raw materials, the target compounds 2-66, 68-76 in Table 1 can be obtained.

[0115] Table 1

[0116]

[0117]

[0118]

[0119]

[0120]

[0121]

[0122]

[0123] Example 77 Determination of DHODH enzyme activity

[0124] The activity of DHODH is determined by cascade reaction to determine its ability to catalyze the oxidation of the natural substrate DHO (dihydroorotic acid) to orotic acid and reduce coenzyme Q at the same time. The activity of DHODH is indirectly detected by measuring the ability of coenzyme Q to reduce the chromogenic substrate DCIP. After the compound and the enzyme reaction mixture are incubated for 5 minutes, the reaction substrate DHO is added to start the reaction, and the enzyme activity is determined by measuring the depletion of DCIP by measuring the absorption at 600nm at a fixed time. Compared with the control group, the inhibition rate of DHODH activity of the compound at different concentrations is calculated, and the half effective inhibitor concentration IC is calculated. 50 The screening results are shown in Table 2.

[0125] Example 78: THP-1 cell antiproliferative activity assay

[0126] THP-1 cells were seeded in 384-well plates (Fisher 142762), with 3,000 cells per well. The next day, graded dilutions of the compounds were added. 72 hours after the addition of the compounds, CellTiter-Lumi (Biyuntian C0068XL) was added to measure the ATP content in the cells, evaluate the cell growth, and calculate the IC of the compounds to inhibit cell growth 50 The screening results are shown in Table 2.

[0127] Example 79: Determination of antiproliferative activity of MV-4-11 cells

[0128] MV-4-11 cells were seeded in 384-well plates (Fisher 142762), with 3000 cells per well. The next day, graded dilutions of the compounds were added. 72 hours after the addition of the compounds, CellTiter-Lumi (Biyuntian C0068XL) was added to measure the ATP content in the cells, evaluate the cell growth, and calculate the IC of the compounds to inhibit cell growth 50 The screening results are shown in Table 2.

[0129] Table 2 Inhibitory activity of the compounds of the present invention on DHODH enzyme, and antiproliferative activity of THP-1 cells and MV-4-11 cells

[0130]

[0131]

[0132] A stands for IC 50 ≤50nM

[0133] B means 50nM≤IC 50 ≤0.5μM

[0134] C stands for IC 50 >0.5μM

[0135] From the data in Table 2, it can be seen that the compounds of the present invention have strong activity in inhibiting DHODH enzyme activity, and the compounds also have strong anti-proliferation activity against tumor cells THP-1 and MV-4-11.

[0136] Although the specific embodiments of the present invention are described above, it should be understood by those skilled in the art that these are only examples, and various changes or modifications may be made to these embodiments without departing from the principles and essence of the present invention. Therefore, the protection scope of the present invention is limited by the appended claims.

Claims

1. A compound represented by the general formula (1), its optical isomer or pharmaceutically acceptable salt: In the general formula (1): n is 0, 1 or 2; X is -CH2-, -O-, -S-, -S(O)-, -S(O)2- or -N(R 5 )-; R 1 is phenyl, pyridyl, pyrazinyl, pyrimidinyl or pyrazolyl, and the phenyl, pyridyl, pyrazinyl, pyrimidinyl and pyrazolyl may be substituted by one or more of the following groups: halogen, OH, NH2, CN, C1-C3 alkyl, C1-C3 alkoxy, halogenated C1-C3 alkyl or halogenated C1-C3 alkoxy; R 2 is H or F; R 3 is C1-C3 alkyl or C3-C8 cycloalkyl; R 4 C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogenated C1-C6 alkyl or C3-C6 cycloalkyl substituted C1-C3 alkyl; R 5 It is H or C1-C3 alkyl.

2. The compound according to claim 1, its optical isomer or pharmaceutically acceptable salt, wherein in the general formula (1), X is -CH2-, -O-, -S-, -S(O)-, -S(O)2-, -NH-, -N(Me)- or -N(Et)-.

3. The compound according to any one of claims 1 to 2, its optical isomer or pharmaceutically acceptable salt, wherein in the general formula (1), R 1 for: Where R a , R b , R c , R d and R e are independently H, F, Cl, Me, Et, OMe, OEt, CHF2, CF3, OH, NH2 or CN.

4. The compound according to any one of claims 1 to 2, its optical isomer or pharmaceutically acceptable salt, wherein in the general formula (1), R 3 For me.

5. The compound according to any one of claims 1 to 2, its optical isomer or pharmaceutically acceptable salt, wherein in the general formula (1), R 4 Me, Et, CF3, 6. The compound, optical isomer or pharmaceutically acceptable salt thereof according to any one of claims 1 to 2, wherein the compound has one of the following structures:

7. A pharmaceutical composition, characterized in that It contains a pharmaceutically acceptable excipient or carrier and the compound according to any one of claims 1 to 6, its optical isomer or pharmaceutically acceptable salt as an active ingredient.

8. Use of the compound according to any one of claims 1 to 6, its optical isomer or pharmaceutically acceptable salt, or the pharmaceutical composition according to claim 7 in the preparation of a medicament for treating DHODH-related diseases.

9. The use according to claim 8, characterized in that The DHODH-related disease is cancer.

10. The use according to claim 9, characterized in that The cancer is a solid tumor or a hematological tumor.

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