CN113429396A

CN113429396A - Five-membered heteroaromatic ring derivative and preparation method and application thereof

Info

Publication number: CN113429396A
Application number: CN202110636378.0A
Authority: CN
Inventors: 黄生宏; 姚任冲
Original assignee: Shenzhen Xianggen Biomedical Co ltd
Current assignee: Shenzhen Xianggen Biomedical Co ltd
Priority date: 2021-06-08
Filing date: 2021-06-08
Publication date: 2021-09-24

Abstract

The five-membered heteroaromatic ring derivative has obvious inhibiting and blocking effects on DHODH, can be used as a DHODH specific inhibitor, can be used for preparing a medicament for treating diseases caused by fungal infection, can be used for preparing an agricultural fungicide, and has wide application prospect.

Description

Five-membered heteroaromatic ring derivative and preparation method and application thereof

Technical Field

The invention relates to the technical field of medicines, in particular to a compound used as a fungal dihydroorotate dehydrogenase (DHODH) inhibitor, a preparation method and application thereof.

Background

Pyrimidine is a key component of RNA and DNA biosynthesis, DHODH catalyzes the 4 th step reaction in the de novo pyrimidine nucleotide synthesis pathway, and pyrimidine nucleotide in cells can be exhausted by inhibiting the activity of the DHODH, so that the growth of the cells is inhibited. Inhibitors against human DHODH may be useful for the treatment of a variety of diseases, including cancer, autoimmune diseases, inflammation, etc., and inhibitors against fungal DHODH may be useful for the treatment of diseases associated with fungal infections.

Currently, the use of many broad spectrum antibiotics and other potent drugs worldwide has led to a substantial increase in the number of patients infected with fungi. There are 29 million Chronic Pulmonary Aspergillosis (CPA) patients, 484 million allergic bronchopulmonary aspergillosis (APBA) patients worldwide, of which about 4 million CPA patients, 49 million APBA patients in china. There are about 6 million patients with invasive fungal infections in europe, and at least 16 million patients with invasive fungal infections in china. Invasive fungal infections, known as hidden killers, are a significant cause of eventual death in patients with tumors, AIDS, organ transplantation, etc., with 5-year mortality rates as high as approximately 80% without immediate diagnosis and long-term antifungal therapy. Strains that cause fungal infections, including aspergillus, sporozoon, etc., remain highly unmet medical needs for therapies that can resist both resistant infections as well as infections with refractory pathogens. Further complicating the treatment of fungal infections, resistance to all antifungal drugs among different fungal pathogens has prompted researchers to design new anti-infective strategies and drugs. Therefore, the development of a DHODH inhibitor with higher efficiency, safety and low side effect is necessary and has great significance.

Disclosure of Invention

The invention mainly solves the technical problem of providing a compound which can effectively inhibit fungi.

In order to solve the technical problems, the invention adopts a technical scheme that:

the invention provides a compound which has a structure shown in a formula I or a tautomer, a mesomer, a racemate, an enantiomer, a diastereomer or a mixture form, a pharmaceutically acceptable hydrate, a solvate or a salt thereof:

R¹selected from substituted or unsubstituted C1-C10 alkyl, substituted or unsubstitutedSubstituted 2-to 10-membered heteroalkyl, substituted or unsubstituted 3-to 12-membered cycloalkyl, substituted or unsubstituted 3-to 12-membered heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl;

R²selected from hydrogen, halogen, cyano, hydroxy, -NH₂A carboxyl group, a substituted or unsubstituted C1-C10 alkyl group, a substituted or unsubstituted 2-to 10-membered heteroalkyl group, a substituted or unsubstituted 3-to 12-membered cycloalkyl group, a substituted or unsubstituted 3-to 12-membered heterocycloalkyl group, a substituted or unsubstituted C2-C10 alkenyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, an acyl group, an ester group, an amide group, a sulfonyl group, a sulfonamide group, a boronic acid ester group;

x, Y, Z are each independently selected from O, S, N, CR³、NR⁴And when Z is NR⁴When X and Y are not CR at the same time³；

W is selected from N, C-H, C-F, C-Cl and C-CH₃、C-CH₂CH₃、C-OH、C-CH₂OH、C-OCH₃；

R³Independently at each occurrence, is selected from hydrogen, halogen, cyano, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted 2-to 10-membered heteroalkyl, substituted or unsubstituted 3-to 12-membered cycloalkyl, substituted or unsubstituted 3-to 12-membered heterocycloalkyl, substituted or unsubstituted C2-C10 alkenyl;

R⁴selected from hydrogen, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted 2-10 membered heteroalkyl, substituted or unsubstituted 3-12 membered cycloalkyl, substituted or unsubstituted 3-12 membered heterocycloalkyl, substituted or unsubstituted C2-C10 alkenyl;

wherein R is¹、R²、R³、R⁴Wherein the substituent is selected from hydrogen, halogen, cyano, C1-C3 alkyl and 2-6-membered heteroalkyl;

further, R¹Selected from substituted or unsubstituted 3-to 6-membered cycloalkyl, substituted or unsubstituted 3-to 6-membered heterocycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted 5-to 9-membered heteroaryl;

further, R¹Selected from the group consisting ofOr unsubstituted phenyl, substituted or unsubstituted 5-6 membered heteroaryl;

in specific embodiments, R¹Is substituted or unsubstituted phenyl, R¹Wherein the substituents are selected from F, Cl, CH₃、CF₃、CHF₂、CH₃O-, preferably F, Cl;

further, R²Selected from hydrogen, halogen, cyano, hydroxy, -NH₂Substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 2-6 membered heteroalkyl, substituted or unsubstituted 3-6 membered cycloalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl;

further, R²Selected from hydrogen, halogen, cyano, hydroxyl, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted C1-C3 alkoxy;

in specific embodiments, R²Selected from hydrogen, F, Cl, cyano, CF₃、CH₃、CH₃O-, preferably F;

further, the air conditioner is provided with a fan,

is selected from

Further, the air conditioner is provided with a fan,

is selected from

In a particular embodiment of the present invention,

is selected from

Preference is given to

Further, W is selected from N, C-H, C-F, C-Cl;

in specific embodiments, W is selected from C-H, C-F, preferably C-H;

further, R³Independently at each occurrence, selected from hydrogen, F, Cl, cyano, CH₃、CF₃、CH₃CH₂-、CH₃O-、CH₃OCH₂CH₂O-；

In specific embodiments, R³Independently at each occurrence, selected from hydrogen, F, Cl, cyano, CH₃Preferably hydrogen, CH₃；

Further, R⁴Selected from hydrogen, CH₃、CH₃CH₂-、CH₃O-、CH₃OCH₂CH₂-；

In specific embodiments, R⁴Selected from hydrogen, CH₃Preferably CH₃；

Further, the compound of the present invention has the structure shown in formula II or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture form, pharmaceutically acceptable hydrate, solvate or salt thereof:

R⁵selected from hydrogen, halogen, cyano, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted 2-10 membered heteroalkoxy, substituted or unsubstituted 3-12 membered cycloalkyl, substituted or unsubstituted 3-12 membered heterocycloalkyl;

further, R⁵Selected from hydrogen, F, Cl, CH₃、CF₃、CHF₂、CH₃O-；

Preferably, R⁵Selected from hydrogen, F, Cl;

more preferablyEarth, R⁵Selected from hydrogen and F.

R²X, Y, Z, W are as defined above;

further, the compound of the present invention has the structure shown in formula III or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture form, pharmaceutically acceptable hydrate, solvate or salt thereof:

wherein R is⁵X, Y, Z are as defined above;

further, the compound is selected from the following structures:

the invention also provides a medicinal composition, which contains the active ingredient of any compound of the invention or the combination of one or more than two of the stereoisomer, the solvate, the hydrate, the pharmaceutically acceptable salt, the ester, the eutectic crystal, the N-oxide, the isotope labeled compound, the metabolite and the prodrug thereof.

The invention also provides the use of the compound of the invention or a stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or co-crystal thereof in the preparation of a DHODH inhibitor.

The invention also provides the use of the compound of the invention or a stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or co-crystal thereof in the preparation of a medicament for treating or preventing fungal infection or a disease caused by fungal infection.

The invention also provides the use of a compound of the invention or a stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or co-crystal thereof in the preparation of a fungicide.

Further, the fungus is selected from one or several organisms of the following genera: absidia, Acremonium, Alternaria, Aspergillus, Bipolaris, Blastomyces, Blumeria, Cladosporium, Coccidioides, Colletotrichium, Curvularia, Encephalitozoon, Epicoccum, Epidermophyton, Exophiala, Exserohilum, Fusarium, Histoplasma, Leptosphaeria, Micropororum, Mycosphaerella, Neurospora, Paecilomyces, Penicillium, Phytophtra, Plasmopara, Pneumocystis, Pyricularia, Pyroluum, Puccinia, Rhizoctonia, Rhizomucor, Edosporium, Trichoplusis, Trichophyton;

further, the fungus is selected from one or several organisms of the following genera: aspergillus, Scedosporium, Fusarium, preferably Aspergillus and/or Scedosporium, more preferably Aspergillus.

Further, the fungus is selected from one or more of a. fumigatus, a. flavus, a. terreus, a. niger, a. lentulus, s.apiospermum, s.prolifican, s.speces.

Further, the fungus is selected from one or more of a. fumigatus, a. flavus, a. terreus, a. niger and s.prolificans.

Further, the diseases caused by fungal infection are selected from diseases caused by systemic fungal infection and diseases caused by surface fungal infection.

Further, the disease of the systemic infectious fungus is selected from the group consisting of pulmonary aspergillosis, allergic bronchopulmonary aspergillosis, systemic aspergillosis, asthma, coccidioidomycosis, paracoccidiomycosis, sporotrichosis, chromoblastomycosis, lacrimomycosis, histoplasmosis, keloid blastomycosis, chromohyphomycosis, disseminated sporotrichosis, fungal colonization of cystic fibrosis, sinusitis.

The disease of surface infection with fungus is selected from tinea, onychomycosis, and tinea pedis.

The invention also provides a preparation method of the compound, which comprises the following steps:

(1) IM-1 and SM-1 are subjected to reaction in the presence or absence of a base and Lewis acid to generate IM-2;

(2) converting IM-2 into M-13 (when Fg and L in the compound are²As such, it should be understood that this step is omitted);

(3) IM-3 and IM-4 are subjected to the action of alkali and/or a condensing agent to generate a compound in the formula I;

L¹selected from halogen, C1-C6 alkoxy, hydroxy, preferably Cl;

L²selected from halogen, hydroxy, preferably Cl, hydroxy;

fg is selected from halogen, C1-C6 alkoxy, hydroxyl, substituted or unsubstituted amino, preferably Cl, methoxy, ethoxy, tert-butoxy, hydroxyl;

R¹、R²w, X, Y, Z are as defined for compounds of the invention;

further, the base in the step (1) is selected from n-butyllithium, sec-butyllithium, lithium diisopropylamide and the like; the Lewis acid is selected from aluminum trichloride, ferric trichloride and the like;

further, in the step (2), the functional group is converted into one or two selected from the following group: (a) hydrolysis of the ester to the acid by the action of a base (e.g., NaOH, LiOH); (b) acid in SOCl₂、POCl₃、PCl₅Action of oxalic acid and the likeDown-conversion to acyl chloride; (c) hydrolyzing the amide into acid under the action of HBr, HCl and the like;

further, the base in the step (3) is selected from triethylamine and N, N-diisopropylethylamine; the condensing agent is selected from 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDCI) or O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium Hexafluorophosphate (HATU).

The pharmaceutical composition containing the compound of the invention or the stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or cocrystal thereof can contain pharmaceutically acceptable auxiliary materials.

As used herein, "pharmaceutically acceptable" is meant to include any material that does not interfere with the effectiveness of the biological activity of the active ingredient and is not toxic to the host to which it is administered.

The pharmaceutically acceptable auxiliary materials are general names of all the additional materials except the main medicine in the medicine, and the auxiliary materials have the following properties: (1) no toxic effect on human body and few side effects; (2) the chemical property is stable and is not easily influenced by temperature, pH, storage time and the like; (3) has no incompatibility with the main drug, and does not influence the curative effect and quality inspection of the main drug; (4) does not interact with the packaging material. The auxiliary materials in the invention include, but are not limited to, a filler (diluent), a lubricant (glidant or anti-adhesion agent), a dispersing agent, a wetting agent, an adhesive, a regulator, a solubilizer, an antioxidant, a bacteriostatic agent, an emulsifier, a disintegrating agent and the like. The binder comprises syrup, acacia, gelatin, sorbitol, tragacanth, cellulose and its derivatives (such as microcrystalline cellulose, sodium carboxymethylcellulose, ethyl cellulose or hydroxypropyl methylcellulose), gelatin slurry, syrup, starch slurry or polyvinylpyrrolidone; the filler comprises lactose, sugar powder, dextrin, starch and its derivatives, cellulose and its derivatives, inorganic calcium salt (such as calcium sulfate, calcium phosphate, calcium hydrogen phosphate, precipitated calcium carbonate, etc.), sorbitol or glycine, etc.; the lubricant comprises superfine silica gel powder, magnesium stearate, talcum powder, aluminum hydroxide, boric acid, hydrogenated vegetable oil, polyethylene glycol and the like; the disintegrating agent comprises starch and its derivatives (such as sodium carboxymethyl starch, sodium starch glycolate, pregelatinized starch, modified starch, hydroxypropyl starch, corn starch, etc.), polyvinylpyrrolidone or microcrystalline cellulose, etc.; the wetting agent comprises sodium lauryl sulfate, water or alcohol, etc.; the antioxidant comprises sodium sulfite, sodium bisulfite, sodium pyrosulfite, dibutylbenzoic acid, etc.; the bacteriostatic agent comprises 0.5% of phenol, 0.3% of cresol, 0.5% of chlorobutanol and the like; the regulator comprises hydrochloric acid, citric acid, potassium (sodium) hydroxide, sodium citrate, and buffer (including sodium dihydrogen phosphate and disodium hydrogen phosphate); the emulsifier comprises polysorbate-80, sorbitan fatty acid, pluronic F-68, lecithin, soybean lecithin, etc.; the solubilizer comprises Tween-80, bile, glycerol, etc. The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention with an acid or base that is suitable for use as a pharmaceutical. The acid base is a generalized Lewis acid base. Suitable acids for forming the salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, etc., organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, phenylmethanesulfonic acid, benzenesulfonic acid, etc.; and acidic amino acids such as aspartic acid and glutamic acid.

The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.

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, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol 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 capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers, e.g., ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide, and oils, especially cottonseed, groundnut, corn germ, olive, castor and sesame oils, or mixtures of such materials, and the like.

In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

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

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 nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of the present 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 which may be required if necessary.

The compounds of the invention can likewise be used in injectable preparations. Wherein the injection is selected from liquid injection (water injection), sterile powder for injection (powder injection) or tablet for injection (refers to impression tablet or machine pressing tablet prepared by aseptic operation method of medicine, and is dissolved with water for injection for subcutaneous or intramuscular injection when in use).

Wherein the powder for injection contains at least an excipient in addition to the above compound. The excipients, which are components intentionally added to a drug in the present invention, should not have pharmacological properties in the amounts used, however, the excipients may aid in the processing, dissolution or dissolution of the drug, delivery by a targeted route of administration, or stability.

"substituted" means that a hydrogen atom in a molecule is replaced by a different atom or molecule.

"element" means the number of skeleton atoms constituting a ring.

The term "one bond" as used herein means only one bond, and may be understood as "none".

The substituent being "═ O" means that multiple substituents on the same atom together form one or more ═ O, e.g. as formed with the atom being substituted

And the like.

"alkyl" refers to an aliphatic hydrocarbon group and to a saturated hydrocarbon group. The alkyl moiety may be a straight chain or branched chain alkyl. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, and the like.

The C1-Cn used in the invention comprises C1-C2 and C1-C3 … … -C1-Cn, wherein n is an integer more than one; the prefix as a substituent denotes the minimum and maximum number of carbon atoms in the substituent, e.g., "C1-C6 alkyl" means a straight or branched chain alkyl group containing one to 6 carbon atoms.

The "hydrocarbon ring" refers to a ring structure having carbon atoms in the ring skeleton, and the carbon atoms may be connected by a single bond or a double bond, so that the hydrocarbon ring includes a saturated hydrocarbon ring, an olefin ring, and an aromatic ring structure.

"alkenyl ring" refers to a group containing a carbon-carbon double bond in the ring backbone.

"heteroalkyl" refers to an alkyl group containing a heteroatom.

"alkenyl" refers to an aliphatic hydrocarbon group having at least one carbon-carbon double bond. The alkenyl groups may be straight-chain or branched.

An "amido" group is a chemical structure having the formula-C (O) NHR or-NHC (O) R, when two linking ends are present, the structure is-C (O) NH (CH)₂) a-or-NHC (O) (CH)₂) a-, wherein R can be selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and the like, and a is a natural number.

"Sulfonyl" is a compound having the formula-S (═ O)₂The chemical structure of R, when having two attached ends, is-S (═ O)₂(CH₂) a-, wherein R can be selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, amino, and the like, and a is a natural number.

"sulfinyl" is a chemical structure having the formula-S (═ O) R, when having two linkers, the structure is-S (═ O) (CH)₂) a-, wherein R can be selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, amino, and the like, and a is a natural number;

"sulfonamido" is of the formula-S (═ O)₂Chemical structure of NHR or-nhc (O) R, when having two linking ends, is-S (═ O)₂NH(CH₂) a-or-NHS (═ O)₂(CH₂) a-, wherein R can be selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and the like, and a is a natural number.

"sulfonamido" is a chemical structure having the formula-S (═ O) NHR or-nhs (O) R, when having two linkagesWhen terminated, the structure is-S (═ O) NH (CH)₂) a-or-NHS (═ O) (CH)₂) a-, wherein R can be selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and the like, and a is a natural number.

A "phosphoryl group" is a chemical structure having the formula-P (═ O) RR', when having two linkers, the structure is-P (═ O) R (CH)₂) a-, wherein R, R' can be independently selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, hydroxy, amino, and the like, and a is a natural number.

"ester group" means a chemical structure having the formula-C (O) OR OR-OC (O) R, when two linkers are present, the structure is-C (O) O (CH)₂) a-or-OC (O) (CH)₂) a-, wherein R can be selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and the like, and a is a natural number.

"acyl" refers to a chemical structure having the formula-C (O) R, when two linkers are present, the structure is-C (O) (CH)₂) a-, wherein R can be selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and the like, and a is a natural number.

"cycloalkyl" refers to a saturated or unsaturated cyclic hydrocarbon substituent such as "C3-C6 cycloalkyl" refers to a cycloalkyl group having 3-6 carbon atoms; in the present invention, the cycloalkyl group may further include a non-aryl structure having an unsaturated bond in the ring skeleton.

"Heterocycloalkyl" refers to a cycloalkyl group containing at least one heteroatom in the ring backbone.

Heteroatoms include, but are not limited to O, S, N, P, Si and the like.

"Ring" refers to any covalently closed structure, including, for example, carbocycles (e.g., aryl or cycloalkyl), heterocycles (e.g., heteroaryl or heterocycloalkyl), aryls (e.g., aryl or heteroaryl), nonaromatic (e.g., cycloalkyl or heterocycloalkyl). The "ring" in the present invention may be a monocyclic ring or a polycyclic ring, and may be a fused ring, a spiro ring or a bridged ring.

Typical heterocycloalkyl groups include, but are not limited to:

"aryl" means a planar ring having a delocalized pi-electron system and containing 4n +2 pi electrons, where n is an integer. The aryl ring may be composed of five, six, seven, eight, nine or more than nine atoms. Aryl groups include, but are not limited to, phenyl, naphthyl, phenanthryl, anthracyl, fluorenyl, indenyl, and the like.

Typical heteroaryl groups include, but are not limited to:

"halogen" or "halo" refers to fluorine, chlorine, bromine or iodine.

As used herein, alkyl, heteroalkyl, cyclic, heterocyclic, amino, ester, carbonyl, amide, sulfonyl, phosphoryl, boronic acid, boronic ester, guanidino, acylguanidino, aryl, heteroaryl, imine and the like are unsubstituted alkyl, heteroalkyl, cyclic, heterocyclic, amino, ester, carbonyl, amide, sulfonyl, phosphoryl, boronic acid, boronic ester, guanidino, acylguanidino, aryl, heteroaryl, imine and the like are substituted alkyl, heteroalkyl, cyclic, heterocyclic, amino, ester, carbonyl, amide, sulfonyl, phosphoryl, boronic acid, boronic ester, guanidino, acylguanidino, aryl, heteroaryl, imine and the like.

Hereinbefore, unless already indicated, the term "substituted" means that the group referred to may be substituted by one or more additional groups each and independently selected from alkyl, cycloalkyl, aryl, carboxy, heteroaryl, heterocycloalkyl, hydroxy, alkoxy, alkylthio, aryloxy, O ═ guanidino, cyano, nitro, acyl, halogen, haloalkyl, amino and the like.

The invention has the beneficial effects that: the invention provides a series of compounds with obvious inhibition effect on DHODH activity, provides a new scheme for treating diseases taking DHODH as a treatment target, such as fungal infection, can be used for preparing medicines for treating related diseases, can also be used for preparing agricultural bactericides, and has wide application prospect.

Detailed Description

The technical solutions of the present invention are described clearly and completely below, and it is obvious that the described embodiments are some, not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The reaction was monitored by Thin Layer Chromatography (TLC) or liquid-mass spectrometry (LC-MS). The developer systems used for silica gel column or thin layer preparative plate purification include, but are not limited to: dichloromethane/methanol system, n-hexane/ethyl acetate system and petroleum ether/ethyl acetate system, the volume ratio of the solvent is adjusted according to different polarities of the compounds, or ammonia water, triethylamine and the like are added for adjustment. The developing solvent system used for reverse phase preparative purification includes but is not limited to (a) phase A: water, phase B: acetonitrile; (b) phase A: water, phase B: methanol.

Unless otherwise specified in the examples, the reaction temperature was room temperature (20 ℃ C. to 30 ℃ C.).

Unless otherwise indicated, reagents used in the examples are commercially available.

In the present invention, the structure of the compound is determined by Mass Spectrometry (MS) and/or nuclear magnetic resonance (M)¹H-NMR) apparatus. The abbreviations have the following meanings:

DMF: n, N-dimethylformamide

THF: tetrahydrofuran (THF)

DIEA: n, N-diisopropylethylamine

Et₃N: triethylamine

DCM: methylene dichloride

HATU: o- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate

DMSO, DMSO: dimethyl sulfoxide

Tolumen. Toluene

1, 4-dioxane: 1,4-dioxane

K₂CO₃: potassium carbonate

Pd(dppf)Cl₂: [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride

NaOH-sodium hydroxide

AlCl₃: aluminium trichloride

t-BuONa: sodium tert-butoxide

LiHMDS: bis-trimethylsilyl amido lithium

diglyme: diethylene glycol dimethyl ether

Intermediate A

Preparation of 4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) aniline (A)

The first step is as follows: preparation of 1- (4-nitrophenyl) piperazine (A-2)

Compound A-1(10.0g,63.5mmol) and piperazine (21.9g,254.2mmol) were added to a reaction flask, dispersed in diethylene glycol dimethyl ether (23mL), the system was heated to 100 ℃ and stirred for 4h, and TLC monitoring indicated completion of the reaction. Cooling the reaction system to about 50 ℃, adding 100mL of water into the system, stirring at room temperature for 1h, then carrying out suction filtration, washing a filter cake with water and petroleum ether in sequence, and drying the obtained solid in vacuum to obtain 12.2g of a product A-2, wherein the yield is as follows: 93 percent.

The second step is that: preparation of 5-fluoro-2- (4- (4-nitrophenyl) piperazin-1-yl) pyrimidine (A-3)

To a reaction flask was added compound A-2(2.0g,9.7mmol), 2-chloro-5-fluoropyrimidine (1.3g,9.8mmol) and K₂CO₃(2.7g,19.5mmol) in diethylene glycol dimethyl ether (12mL) and the system heated to 100 ℃ stirring the reaction overnight with LCMS monitoring showing completion of the reaction. Cooling the reaction system to room temperature, adding a proper amount of water, stirring, then carrying out suction filtration, washing a filter cake with water and petroleum ether in sequence, and carrying out vacuum drying on the obtained solid to obtain 2.3g of a product A-3 with yield: 79 percent.

The third step: preparation of 4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) aniline (A)

Compound A-3(1.2g,4.0mmol) was added to a reaction flask, dissolved in methanol (30mL), and then Pd/C (240mg) was added, the reaction system was replaced with hydrogen gas, the reaction solution was stirred overnight at room temperature under a hydrogen atmosphere, and TLC monitoring indicated completion of the reaction. Suction filtration, reduced pressure concentration of the filtrate, purification of the crude product through silica gel column to obtain 1.0g of product a, yield: 92 percent.

Example 1

Preparation of N- (4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) phenyl) -2- (1-methyl-4-phenyl-1H-pyrazol-5-yl) -2-oxyacetamide (T-1)

The first step is as follows: preparation of 1-methyl-4-phenyl-1H-pyrazole (1-2)

To a reaction flask were added compound 1-1(2.0g,9.6mmol), phenylboronic acid (1.3g,10.7mmol), and K₂CO₃(4.0g,28.9mmol), dispersed in 1,4-dioxane (25mL) and water (5mL), the air in the reaction flask was replaced with nitrogen, and Pd (dppf) Cl was added₂(366mg,0.5mmol), the system was heated to 80 ℃ under nitrogen and the reaction stirred overnight, TLC monitoring showed completion of the reaction. Cooling the reaction system to room temperature, adding appropriate amount of water, extracting with ethyl acetate, washing the organic phase with saturated brine, and anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product with silica gel column to obtain 1.0g of product 1-2, yield: 66 percent.

The second step is that: preparation of methyl 2- (1-methyl-4-phenyl-1H-pyrazol-5-yl) -2-oxoacetate (1-3)

Adding the compound 1-2(880mg,5.56mmol) into a reaction flask, dissolving the compound in anhydrous tetrahydrofuran (20mL), replacing air in the reaction flask with nitrogen, cooling the system to-70 ℃, then dropwise adding n-butyllithium solution (2.5M in hexane,2.5mL,6.25mmol), stirring the system at-70 ℃ for reaction for 1h, dropwise adding a solution of dimethyl oxalate (2.0g,16.94mmol) in anhydrous tetrahydrofuran (5mL), controlling the temperature of the system to be below-65 ℃ during dropwise adding, then slowly returning to room temperature and stirring the reaction overnight, and monitoring by TLC to show that the reaction is complete. Adding saturated NH into the reaction system₄The reaction was quenched with Cl solution, extracted with ethyl acetate, the organic phase washed with saturated brine, anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product through silica gel column to obtain 615mg of product 1-3, yield: 45 percent.

The third step: preparation of 2- (1-methyl-4-phenyl-1H-pyrazol-5-yl) -2-oxoacetic acid (1-4)

Compound 1-3(250mg,1.02mmol) was added to the reaction flask, dissolved in methanol (5mL) and water (1mL), and sodium hydroxide (61mg,1.53mmol) was added under ice water bath, the system was allowed to return to room temperature and stirred for 2h, and TLC monitoring indicated completion of the reaction. Adding appropriate amount of water into the reaction system, adjusting the system to acidity with 1N diluted hydrochloric acid solution, extracting with ethyl acetate, and extracting the organic phase with anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product through silica gel column to obtain 175mg of product 1-4, yield: 74 percent.

The fourth step: preparation of N- (4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) phenyl) -2- (1-methyl-4-phenyl-1H-pyrazol-5-yl) -2-oxyacetamide (T-1)

Compound 1-4(20mg,0.087mmol) was added to the reaction flask, dissolved in dichloromethane (2mL), the system was cooled to 0 ℃ under nitrogen, a solution of oxalyl chloride (11mg,0.087mmol) in dichloromethane (1mL) was added dropwise to the system, and the system was stirred at 0 ℃ for 1 hour. The solution prepared above was added dropwise to a solution containing Compound A (24mg,0.088mmol) and Et under ice bath₃In a reaction flask containing a solution of N (27mg,0.267mmol) in dichloromethane (2mL), the reaction was allowed to slowly return to room temperature and the reaction was stirred for 3h, and TLC monitoring indicated completion of the reaction. Adding appropriate amount of water into the reaction system to quench the reaction, extracting with dichloromethane, washing the organic phase with saturated brine, and washing with anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product with thin-layer silica gel plate to obtain 6mg of product T-1 with yield: 14 percent.

LC-MS(ESI)m/z(M+H)⁺：486.2

¹H NMR(400MHz,DMSO-d₆)δ10.63(s,1H),8.49(s,2H),7.77(s,1H),7.33-7.29(m,2H),7.27-7.16(m,5H),6.91-6.86(m,2H),4.09(s,3H),3.83-3.78(m,4H),3.18-3.12(m,4H).

Example 2

Preparation of N- (4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) phenyl) -2- (5-methyl-3-phenylfuran-2-yl) -2-oxyacetamide (T-2)

The first step is as follows: preparation of 2-methyl-4-phenyl furan (2-2)

Acetone (641mg,11.04mmol) is added into a reaction bottle, dissolved in toluene (45mL), the system is cooled to-78 ℃ under the protection of nitrogen, LiHMDS (1.06M in THF/ethylbenzene,11mL,11.66mmol) is slowly dripped into the system, the reaction system is stirred for 1.5h at-78 ℃ after the dripping is finished, then a THF (10mL) solution of a compound 2-1(2.0g,11.05mmol) is dripped into the system, the temperature of the system is controlled below-70 ℃ during the dripping, the dripping is finished, the reaction system is slowly returned to room temperature and stirred overnight, and the TLC monitoring shows that the reaction is complete. Adding saturated NH into the reaction system₄The reaction was quenched with aqueous Cl, extracted with dichloromethane, the organic phase washed with saturated brine, anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the obtained crude product through silica gel column to obtain 200mg of product 2-2, yield: 13 percent.

The second step is that: preparation of 2- (5-methyl-3-phenylfuran-2-yl) -2-oxoacetyl chloride (2-3)

Compound 2-2(70mg,0.44mmol) was added to the reaction flask, dissolved in dichloromethane (3mL), replaced with nitrogen, cooled to 0 ℃ on an ice bath, and then a solution of oxalyl chloride (68mg,0.53mmol) in dichloromethane (1mL) was added dropwise to the system, after completion of the dropwise addition, the reaction was stirred under nitrogen for 1 hour, and TLC monitoring indicated complete reaction of the starting material. The reaction system was concentrated under reduced pressure at low temperature to give 110mg of crude 2-3, which was used in the next reaction without purification.

The third step: preparation of N- (4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) phenyl) -2- (5-methyl-3-phenylfuran-2-yl) -2-oxyacetamide (T-2)

Compound A (110mg,0.40mmol) was added to the reaction flask, dissolved in dichloromethane (5mL), and thenDIEA (155mg,1.20mmol) was then added, replaced with nitrogen, cooled to 0 ℃ on an ice bath, and a solution of compound 2-3(110mg, 0.44mmol) in dichloromethane (3mL) was added dropwise to the reaction, after which the reaction was allowed to warm slowly to room temperature and stirred overnight, as monitored by LCMS indicating completion of the reaction. Adding water to the reaction system to quench the reaction, extracting with dichloromethane, washing the organic phase with saturated brine, and washing with anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product with thin layer silica gel plate to obtain 90mg of product T-2, yield: two steps 42%.

LC-MS(ESI)m/z(M+H)⁺：486.2

¹H NMR(400MHz,DMSO-d₆)δ10.59(s,1H),8.50(s,2H),7.70-7.66(m,2H),7.45-7.38(m,5H),7.00-6.95(m,2H),6.78(s,1H),3.86-3.81(m,4H),3.20-3.16(m,4H),2.44(s,3H).

Example 3

Preparation of 2- (1, 2-dimethyl-4-phenyl-1H-pyrrol-3-yl) -N- (4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) phenyl) -2-oxyacetamide (T-3)

The first step is as follows: preparation of 5-methyl-3-phenyl-1H-pyrrole-2-carboxylic acid ethyl ester (3-2)

Compound 3-1(3.5g,21.58mmol) and glycine ethyl ester hydrochloride (30.0g,214.93mmol) were added to a reaction flask, dissolved in dry DMF (125mL), replaced with nitrogen, and the reaction was heated to 150 ℃ under nitrogen and stirred for two days. Cooling the reaction system to room temperature, adding appropriate amount of water, extracting with ethyl acetate, washing the organic phase with water and saturated brine successively, and removing anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product with silica gel column to obtain 1.1g of product 3-2, yield: 22 percent.

The second step is that: preparation of 1, 5-dimethyl-3-phenyl-1H-pyrrole-2-carboxylic acid ethyl ester (3-3)

To a sealed reaction tube was added compound 3-2(350mg,1.53mmol), dissolved in DMF (5mL), and K was added₂CO₃(633mg,4.58mmol) and methyl iodide(633mg,4.46mmol) and the system was sealed and heated to 80 ℃ and the reaction stirred for 6h, monitored by TLC to show complete reaction of the starting materials. Cooling the reaction system to room temperature, adding appropriate amount of water, extracting with ethyl acetate, washing the organic phase with water and saturated brine successively, and removing anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product with silica gel column to obtain 335mg of product 3-3, yield: 90 percent.

The third step: preparation of 4- (2-methoxy-2-oxoacetyl) -1, 5-dimethyl-3-phenyl-1H-pyrrole-2-carboxylic acid ethyl ester (3-4)

Add Compound 3-3(300mg,1.23mmol) to the reaction flask, dissolve in chloroform (15mL), add AlCl₃(811mg,6.08mmol) was replaced with nitrogen, the reaction was heated to 50 ℃ under nitrogen blanket, and then oxalyl chloride monomethyl ester (452mg,3.69mmol) was added and the reaction stirred at this temperature for 6 h. Cooling the reaction system to room temperature, adding a proper amount of water to quench the reaction, extracting with dichloromethane, washing the organic phase with saturated brine, and obtaining anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the obtained crude product through silica gel column to obtain 60mg of product 3-4, yield: 15 percent.

The fourth step: preparation of 4- (carboxycarbonyl) -1, 5-dimethyl-3-phenyl-1H-pyrrole-2-carboxylic acid (3-5)

The reaction flask was charged with compound 3-4(60mg,0.18mmol), dispersed in ethanol (1mL) and water (1mL), NaOH (73mg,1.83mmol) was added, the system was heated to reflux and stirred overnight, and LCMS monitoring indicated complete reaction of the starting material. Cooling the reaction system to room temperature, concentrating under reduced pressure to remove the organic solvent, adding a proper amount of water, adjusting the pH value of the system to 2 by using concentrated hydrochloric acid in an ice water bath, generating a solid, performing suction filtration, washing a filter cake by using water, and performing vacuum drying to obtain a product of 35mg, namely 3-5, wherein the yield is as follows: 67%.

The fifth step: preparation of 2- (1, 2-dimethyl-4-phenyl-1H-pyrrol-3-yl) -2-oxoacetic acid (3-6)

The compound 3-5(30mg,0.104mmol) was added to the reaction flask, dissolved in DMF (3mL), replaced with nitrogen, and the system was heated to 150 ℃ and stirred for reaction for 4 h. Cooling the reaction system to room temperature, adding appropriate amount of water, extracting with ethyl acetate, washing the organic phase with water and saturated brine successively, and removing anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product with thin layer silica gel plate to obtain 20mg of product 3-6 with yield: 79 percent.

And a sixth step: preparation of 2- (1, 2-dimethyl-4-phenyl-1H-pyrrol-3-yl) -N- (4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) phenyl) -2-oxyacetamide (T-3)

To a reaction flask, compound 3-6(20mg,0.082mmol) was added, dissolved in dichloromethane (2mL), and Et was added₃N (24mg,0.237mmol) and HATU (91mg,0.239mmol) were stirred at room temperature for 1h, then Compound A (26mg,0.095mmol) was added and the reaction stirred at room temperature overnight with TLC monitoring showing completion of the reaction. Quenching the reaction with appropriate amount of water, extracting with dichloromethane, washing the organic phase with saturated brine, anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product with thin-layer silica gel plate to obtain 8mg of product T-3 with yield: 20 percent.

LC-MS(ESI)m/z(M+H)+：499.2

¹H NMR(400MHz,DMSO-d6)δ9.66(s,1H),9.50(s,1H),8.48(s,2H),7.37-7.33(m,4H),7.32-7.28(m,1H),7.27-7.22(m,2H),6.89(d,J＝8.8Hz,2H),3.83-3.78(m,4H),3.66(s,3H),3.15-3.10(m,4H),2.59(s,3H).

Example 4

Preparation of 2- (4, 5-dimethyl-2-phenyl-1H-pyrrol-3-yl) -N- (4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) phenyl) -2-oxyacetamide (T-4)

The first step is as follows: preparation of 3-acetyl-3-methyl-1-phenylpentane-1, 4-dione (4-2)

To a reaction flask were added compound 2-1(1.0g,5.02mmol), compound 4-1(688mg,6.03mmol) and K₂CO₃(2.1g,15.19mmol) in DMF (5mL), the reaction was heated to 50 ℃ and stirred for 3h, and TLC monitoring indicated complete reaction. Cooling the reaction system to room temperature, adding appropriate amount of water, extracting with ethyl acetate, washing the organic phase with water and saturated brine successively, and removing anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product through silica gel column to obtain 650mg of product 4-2, yield: 56 percent.

The second step is that: preparation of 2, 3-dimethyl-5-phenyl-1H-pyrrole (4-3)

To a sealed reaction flask was added compound 4-2(620mg,2.67mmol) dissolved in methanolic ammonia (7Min CH)₃OH, 20mL), the reaction was sealed and stirred at room temperature overnight, TLC showed the starting material was completely reacted. The solvent was removed by concentration under reduced pressure, and the resulting crude product was purified by silica gel column to give 355mg of the product 4-3, yield: 78 percent.

The third step: preparation of 2- (4, 5-dimethyl-2-phenyl-1H-pyrrol-3-yl) -N- (4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) phenyl) -2-oxyacetamide (T-4)

Compound 4-3(27mg,0.16mmol) was added to the reaction flask, dissolved in dichloromethane (2mL), the system was cooled to 0 ℃ under nitrogen, oxalyl chloride (22mg,0.17mmol) was added to the system, and the system was slowly returned to room temperature and stirred for reaction for 1 hour. The solution prepared above was added dropwise to a mixture of Compound A (35mg,0.13mmol) and Et under ice bath₃In a reaction flask containing a solution of N (26mg,0.26mmol) in dichloromethane (2mL), the reaction was slowly returned to room temperature and the reaction was stirred for 1h, and TLC monitoring indicated completion of the reaction. Adding appropriate amount of water into the reaction system to quench the reaction, extracting with dichloromethane, washing the organic phase with saturated brine, and washing with anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product with thin layer silica gel plate to obtain 40mg of product T-4, yield: and 63 percent.

LC-MS(ESI)m/z(M+H)+：499.2

¹H NMR(400MHz,DMSO-d6)δ11.43(s,1H),10.24(s,1H),8.48(s,2H),7.40-7.36(m,2H),7.26-7.18(m,5H),6.88-6.84(m,2H),3.83-3.78(m,4H),3.15-3.10(m,4H),2.15(s,3H),2.11(s,3H).

Example 5

Preparation of N- (4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) phenyl) -2-oxo-2- (1,4, 5-trimethyl-2-phenyl-1H-pyrrol-3-yl) acetamide (T-5)

The first step is as follows: preparation of 1,2, 3-trimethyl-5-phenyl-1H-pyrrole (5-1)

The compound 4-3(320mg,1.87mmol) and 18-crown-6 (1.6g,6.05mmol) were added to a reaction flask, dissolved in toluene (20mL), and t-BuOK (630mg,5.61mmol) was added to the system under ice-water bath, followed by dropwise addition of iodomethane (320mg,1.87mmol), after which the system was returned to room temperature and stirred for 1h, TLC showed complete reaction of the starting materials. Adding appropriate amount of water into the reaction system to quench the reaction, extracting with ethyl acetate, washing the organic phase with water and saturated brine successively, and anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product through silica gel column to obtain 143mg of product 5-1, yield: 41 percent.

The second step is that: preparation of N- (4- (4- (5-fluoropyrimidin-2-yl) piperazin-1-yl) phenyl) -2-oxo-2- (1,4, 5-trimethyl-2-phenyl-1H-pyrrol-3-yl) acetamide (T-5)

Compound 5-1(30mg,0.16mmol) was added to a reaction flask, dissolved in dichloromethane (2mL), the system was cooled to 0 ℃ under nitrogen, oxalyl chloride (22mg,0.17mmol) was added to the system, and the system was slowly returned to room temperature and stirred for reaction for 1 hour. The solution prepared above was added dropwise to a mixture of Compound A (35mg,0.13mmol) and Et under ice bath₃In a reaction flask containing a solution of N (26mg,0.26mmol) in dichloromethane (2mL), the reaction was slowly returned to room temperature and the reaction was stirred for 1h, and TLC monitoring indicated completion of the reaction. Adding appropriate amount of water into the reaction system to quench the reaction, extracting with dichloromethane, washing the organic phase with saturated brine, and washing with anhydrous Na₂SO₄Drying, concentrating under reduced pressure, and purifying the crude product with thin layer silica gel plate to obtain 40mg of product T-5, yield: 61 percent.

LC-MS(ESI)m/z(M+H)+：513.2

¹H NMR(400MHz,DMSO-d₆)δ10.03(s,1H),8.48(s,2H),7.27-7.19(m,5H),7.08-7.03(m,2H),6.83-6.78(m,2H),3.83-3.78(m,4H),3.26(s,3H),3.14-3.09(m,4H),2.19(s,3H),2.18(s,3H).

Examples 6 to 7: preparation of Compounds T-6 to T-7

By using the method for preparing the compound T-4, the compounds T-6 to T-7 are prepared by adopting different raw materials (cyclopropylammonia and ethanolamine are respectively used for replacing the ammonia methanol solution in the second step in the embodiment 4), and the structural formula, LC-MS and the formula thereof¹The H-NMR data are shown in Table 1.

Table 1: structures, LC-MS and 1H-NMR data of examples 6 to 7

Test example 1: determination of Minimum Inhibitory Concentration (MIC)

1.1 sporulation

A glycerol stock of A.fumigatus (ATCC MYA-4609/AF 293/CBS 101355) strain was used to make new streaks on Sabouraud Dextrose Agar (SDA). Culturing at 35 deg.C and 40-70% humidity for 72 hr.

1.2 preparation of the culture Medium

RPMI 1640: a pack of RPMI1640 powder was dissolved in 1L of purified water and mixed well, 0.165M MOPS was added, the pH was adjusted to 7.0, and sterilized by filtration through a 0.22um membrane. Storing at 4 deg.C for no more than 3 months.

Physiological saline: 9g of sodium chloride powder are dissolved in 1L of purified water and mixed well and autoclaved at 121 ℃ for 30 minutes. Stored at Room Temperature (RT) for no more than 1 week.

1.3 preparation of test boards

1.3.1 preparation of stock solutions

Stock solutions of test compound and one control compound were dissolved in dimethyl sulfoxide to 12.8 mg/mL.

1.3.2100 preparation of working solution

The highest concentration of each compound was 0.8mg/mL (e.g., 150. mu.L of dimethyl sulfoxide +10uL of 12.8mg/mL compound/control). Column 1 was filled with 40. mu.L of compound and columns 2 to 11 were filled with 20. mu.L of dimethyl sulfoxide. mu.L of compound was transferred from column 1 to column 2 and the mixture was diluted. The above procedure was repeated until column 11 to give 2 × serial dilutions.

1.3.3 preparation of test boards

mu.L of the above 2X series of dilutions were injected into each well of the corresponding test plate using a multichannel pipette.

1.4 preparation of inoculum

5ml of saline was dropped on the A.fumigatus SDA plate. The spores on the tray surface were carefully wiped off with an L-shaped coating bar and the suspension was then transferred to a sterile tube. After shaking to mix well, the tube was allowed to stand for 5-10 minutes, and then the number of spores in the suspension was counted by a hemocytometer and checked for purity. Adjusting the inoculum concentration to 0.2-2.5 × 10⁴spores/mL.

1.5 addition of fungi

Then 198 μ L of diluted inoculum was injected into each well of the corresponding assay plate using a multi-channel pipette.

1.6 hatching

The culture was carried out at 35 ℃ for 48 hours.

1.7 MIC assay

The lowest concentration of each sample that completely/significantly (e.g., 80% inhibition) inhibited visible growth of the fungus after incubation was recorded as the MIC. To facilitate scoring of the presence or absence of growth on the wall, a magnifying glass device was used. The 96-well microplate tested was photographed and the 530nm optical density was read with a microplate reader (Tecan Spark). .

The MIC results for the compounds of the invention are shown in table 2.

Table 2: MIC results for Compounds

Examples	AF293 MIC(μg/mL)
		2	0.125
3	1
		4	0.125
5	0.031

The compound of the invention shows stronger inhibitory activity to aspergillus fumigatus (ATCC MYA-4609/AF 293/CBS 101355) in a test for determining Minimum Inhibitory Concentration (MIC).

Test example 2: pharmacokinetic testing

Each test compound was administered orally (10 mg/kg, 3 per group) in a single dose to SD rats for pharmacokinetic studies, and the test compound was dissolved with 5% DMSO + 10% solutol + 85% saline and formulated into a colorless, transparent, clear dosing solution after vortexing for 1-2min and sonicating for 5-10 min. Animals were fasted overnight prior to oral administration and returned to chow 4 hours after administration. After the oral administration of the SD rat, pharmacokinetic samples are collected by orbital blood collection, and the collection time points are as follows: at 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h and 24h after administration, 3 whole blood samples are collected at each time point, and the collection amount is about 0.2-0.3 mL. Immediately after the blood sample was collected, the blood plasma was centrifuged within 15 minutes on ice (centrifugation conditions: 8000 rpm, 1 minute, room temperature). The collected plasma was stored at-20 ℃ before analysis. And (3) taking 20 mu L of plasma sample into a 0.5mL EP tube, adding 200 mu L of working internal standard solution (blank without the internal standard and adding the solvent with the same volume), vortex mixing for 3min, centrifuging at 13500 rpm for 10min, taking 100 mu L of supernatant, and analyzing by LC-MS/MS injection.

The results of the pharmacokinetic testing of some of the compounds of the invention are shown in table 3 below:

table 3: results of pharmacokinetic testing of a portion of the Compounds of the invention

Note: cmax: maximum compound concentration; AUC: exposure amount; t1/2: a half-life; tmax: time to Cmax.

The compounds of the invention have good exposure and compound concentration maxima in pharmacokinetic experiments.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

1. A five-membered heteroaromatic ring derivative having the structure of formula I or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, pharmaceutically acceptable hydrate, solvate, or salt thereof:

R¹selected from substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted 2-10 membered heteroalkyl, substituted or unsubstituted 3-12 membered cycloalkyl, substituted or unsubstituted 3-12 membered heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl;

wherein R is¹、R²、R³、R⁴Wherein the substituent is selected from hydrogen, halogen, cyano, C1-C3 alkyl, and 2-6 membered heteroalkyl.

2. The five-membered heteroaromatic ring derivative of claim 1, wherein R is¹Selected from substituted or unsubstituted 3-to 6-membered cycloalkyl, substituted or unsubstituted 3-to 6-membered heterocycloalkyl, substituted or unsubstituted phenyl, and substituted or unsubstituted 5-to 9-membered heteroaryl.

3. The five-membered heteroaromatic ring derivative of claim 2, wherein R is¹Selected from substituted or unsubstituted phenyl, substituted or unsubstituted 5-6 membered heteroaryl.

4. The five-membered heteroaromatic ring derivative of claim 3, wherein R¹Is substituted or unsubstituted phenyl, R¹Wherein the substituents are selected from F, Cl, CH₃、CF₃、CHF₂、CH₃O-。

5. The five-membered heteroaromatic ring derivative according to any one of claim 1, wherein R²Selected from hydrogen, halogen, cyano, hydroxy,-NH₂Substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 2-to 6-membered heteroalkyl, substituted or unsubstituted 3-to 6-membered cycloalkyl, substituted or unsubstituted 3-to 6-membered heterocycloalkyl.

6. The five-membered heteroaromatic ring derivative of claim 5, wherein R²Selected from hydrogen, halogen, cyano, hydroxyl, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted C1-C3 alkoxy.

7. The five-membered heteroaromatic ring derivative of claim 5, wherein R²Selected from hydrogen, F, Cl, cyano, CF₃、CH₃、CH₃O-。

8. The five-membered heteroaromatic ring derivative according to claim 1,

is selected from

R³Independently at each occurrence, selected from hydrogen, F, Cl, cyano, CH₃、CF₃、CH₃CH₂-、CH₃O-、CH₃OCH₂CH₂O-；R⁴Selected from hydrogen, CH₃、CH₃CH₂-、CH₃O-、CH₃OCH₂CH₂-。

9. The five-membered heteroaromatic ring derivative of claim 1, wherein W is selected from the group consisting of N, C-H, C-F, C-Cl.

10. The five-membered heteroaromatic ring derivative according to claim 1, wherein said five-membered heteroaromatic ring derivative has the structure of formula II or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, pharmaceutically acceptable hydrate, solvate or salt thereof:

R⁵selected from hydrogen, halogen, cyano, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted 2-10 membered heteroalkoxy, substituted or unsubstituted 3-12 membered cycloalkyl, substituted or unsubstituted 3-12 membered heterocycloalkyl.

11. The five-membered heteroaromatic ring derivative of claim 10, wherein R⁵Selected from hydrogen, F, Cl, CH₃、CF₃、CHF₂、CH₃O-。

12. The five-membered heteroaromatic ring derivative according to claim 1, wherein said five-membered heteroaromatic ring derivative has the structure of formula III or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, pharmaceutically acceptable hydrate, solvate or salt thereof:

13. the five-membered heteroaromatic ring derivative according to claim 1, selected from the group consisting of the following structures:

14. a pharmaceutical composition, wherein the active ingredient of the pharmaceutical composition is selected from the five-membered heteroaromatic ring derivatives of any one of claims 1 to 13, or a combination of one or more of stereoisomers, solvates, hydrates, pharmaceutically acceptable salts, esters, co-crystals, N-oxides, isotopically labeled compounds, metabolites and prodrugs thereof.

15. Use of a five-membered heteroaromatic ring derivative according to any of claims 1 to 13 or a stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or co-crystal thereof for the preparation of a DHODH inhibitor.

16. Use of a five-membered heteroaromatic ring derivative according to any one of claims 1 to 13 or a stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or co-crystal thereof for the preparation of a medicament for the treatment or prevention of a fungal infection or a disease caused by a fungal infection.

17. Use of a five-membered heteroaromatic ring derivative according to claim 16, wherein said fungus is selected from one or more organisms of the genera: absidia, Acremonium, Alternaria, Aspergillus, Bipolaris, Blastomyces, Blumeria, Cladosporium, Coccidioides, Colletotrichium, Curvularia, Encephalitozoon, Epicoccum, Epidermophyton, Exophiala, Exserohilum, Fusarium, Histoplasma, Leptosphaeria, Micropororum, Mycosphaerella, Neurospora, Paecilomyces, Penicillium, Phytophtra, Plasmopara, Pneumocystis, Pyricularia, Pyrhium, Puccinia, Rhizoctonia, Rhizomucor, Edosporium, Photospira, Trichophyllum, Trichophyton, and Trichophyton.

18. Use of a five-membered heteroaromatic ring derivative according to claim 16 wherein said disease caused by fungal infection is selected from the group consisting of systemic fungal infection and superficial fungal infection.

19. Use of a five-membered heteroaromatic ring derivative according to claim 18 wherein said systemically infected fungal disease is selected from the group consisting of pulmonary aspergillosis, allergic bronchopulmonary aspergillosis, systemic aspergillosis, asthma, coccidioidomycosis, paracoccidiomycosis, sporotrichosis, chromoblastomycosis, lachnomycosis, histoplasmosis, keloid blastomycosis, chromosporotrichosis, disseminated sporotrichosis, fungal colonization of cystic fibrosis, sinusitis.

20. Use of a five-membered heteroaromatic ring derivative according to claim 18 wherein said superficial fungal infection is selected from the group consisting of ringworm, onychomycosis, and tinea pedis.

21. A process for the preparation of a five-membered heteroaromatic ring derivative, characterized in that the five-membered heteroaromatic ring derivative is a five-membered heteroaromatic ring derivative according to any one of claims 1 to 13, comprising the steps of:

(2) converting IM-2 into M-13 through one-step or multi-step functional groups;

L¹selected from halogen, C1-C6 alkoxy and hydroxyl;

L²selected from halogen, hydroxy, preferably Cl, hydroxy;

fg is selected from halogen, C1-C6 alkoxy, hydroxyl, substituted or unsubstituted amino.