Substituted naphthyl methyl oxadiazole-aniline compound and application thereof

Abstract

The invention belongs to the technical field of medicines, and discloses a substituted naphthyl methyl oxadiazole-aniline compound, a preparation method thereof, a pharmaceutical composition taking the compound as an active component, and application of the substituted naphthyl methyl oxadiazole-aniline compound and the pharmaceutical composition in treatment and / or prevention of infectious diseases caused by coronavirus. Specifically, the invention relates to a compound shown as a formula (I) and an isomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition containing the compound disclosed by the invention, and R1, R2, R3, R4, m and n are described in the specification. The invention aims to prepare a new compound with anti-coronavirus activity, and the new compound can be used as a potential new drug for treatment or prophylactic treatment of infectious diseases caused by viruses, especially diseases caused by coronaviruses. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medical technology. In particular, it relates to substituted naphthylmethyl oxadiazole-aniline compounds represented by general formula (I), preparation methods thereof, pharmaceutical compositions with the compounds as active ingredients, and their use in treating and / or preventing infectious diseases caused by coronaviruses, especially the new coronavirus (SARS-CoV-2). Background Art

[0002] Viral infections have been a continuous threat to human life and health. Currently, there is still a lack of effective antiviral drugs for most viruses. COVID-19 is an infectious disease caused by the new coronavirus SARS-CoV-2. It is highly secretive and infectious, causing great harm to social public health security. The efficacy of existing antiviral drugs and traditional Chinese medicines is not certain. Supportive and symptomatic treatments are still the main treatments in clinical practice. There is currently no specific drug for this virus. Therefore, it is of great significance to find targeted, efficient and low-toxic drugs for the treatment of this virus based on the structure of coronavirus. SARS-CoV-2 encodes 16 non-structural proteins. As the protein structures are analyzed one by one, drug development technologies based on the druggable target structures of coronavirus have begun to be widely adopted. Among them, the representative progress is the successful launch of Paxlovid, an inhibitor targeting Mpro. Papain-like protease (PLpro) is a non-structural protein containing 300 amino acids. It is responsible for the mature shearing of viral precursor proteins and is an essential protein for viral replication in cells. In addition, PLpro is also involved in key processes such as viral antagonism of host innate immunity. The multiple functions of PLpro in the virus life cycle make it an important research and development target for drugs against the new coronavirus. GRL0617 is a lead compound discovered after screening and structural optimization of SARS-CoV PLpro. It has high inhibitory activity and target enzyme affinity. The three-dimensional structure of PLpro has been resolved, and the crystal structure of its complex with GRL0617 has also been reported. These research results have laid the foundation for the development of new PLpro inhibitors. Summary of the invention

[0003] The technical problem to be solved by the present invention is to provide a substituted naphthylmethyl oxadiazole-aniline compound with a novel structure and anti-coronavirus, especially coronavirus (SARS-CoV / SARS-CoV-2) effect and PLpro activity inhibition. The present invention finds that the substituted naphthylmethyl oxadiazole-aniline compound has a strong PLpro inhibitory activity and anti-coronavirus SARS-CoV / SARS-CoV-2 effect, and has a low cytotoxic effect and a metabolic stability better than GRL0617, and can be used for infectious diseases caused by viruses, especially the treatment or preventive treatment of diseases caused by the new coronavirus. The present invention is completed based on the above findings. SUMMARY OF THE INVENTION

[0005] To this end, the present invention provides, in a first aspect, a compound represented by general formula (I) and an isomer thereof, or a pharmaceutically acceptable salt thereof,

[0006]

[0007] in,

[0008] R1 is hydrogen, methyl, ethyl, propyl, isopropyl;

[0009] R2 is independently selected from hydrogen, F, Cl, Br, hydroxy, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, n represents 1, 2, 3 or 4 substituents;

[0010] R3 is hydrogen, methyl, ethyl, propyl;

[0011] m is 0 or 1;

[0012] R4 is substituted or unsubstituted C3-C7 cycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted thienyl, substituted or unsubstituted furanyl, substituted or unsubstituted spiro[3,3]heptyl;

[0013] The substituents in R4 may be selected from the following groups: F, Cl, Br, hydroxyl, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, vinyl, ethynyl, hydroxymethyl, carboxyl, sulfonic acid, boric acid, methylthio, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxycarbonyl, C1-C4 alkoxysulfonyl, C1-C4 alkylamino.

[0014] In a preferred embodiment, the compound has the structural formula as shown in (II):

[0015]

[0016] in,

[0017] R1 is hydrogen or methyl;

[0018] R2 is independently selected from hydrogen, F, Cl, Br, hydroxy, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, n represents 1, 2, 3 or 4 substituents;

[0019] m is 0 or 1;

[0020] R4 is substituted or unsubstituted C3-C7 cycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted thienyl, substituted or unsubstituted furanyl, substituted or unsubstituted spiro[3,3]heptyl;

[0021] The substituents in R4 may be selected from the following groups: F, Cl, Br, hydroxyl, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, vinyl, ethynyl, hydroxymethyl, carboxyl, sulfonic acid, boric acid, methylthio, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxycarbonyl, C1-C4 alkoxysulfonyl, C1-C4 alkylamino.

[0022] In some aspects, the compound described by formula (II), wherein:

[0023] R1 is hydrogen or methyl;

[0024] R2 is independently selected from hydrogen, F, Cl, Br, hydroxy, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, n represents 1, 2, 3 or 4 substituents;

[0025] m is 0;

[0026] R4 is substituted or unsubstituted C3-C7 cycloalkyl, substituted or unsubstituted spiro[3,3]heptyl;

[0027] The substituents in R4 may be selected from the following groups: F, Cl, Br, hydroxyl, amino, nitro, cyano, hydroxymethyl, carboxyl, sulfonic acid, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxycarbonyl.

[0028] In other aspects, the compound described by formula (II), wherein:

[0029] R1 is hydrogen or methyl;

[0030] R2 is independently selected from hydrogen, F, Cl, Br, hydroxy, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, n represents 1, 2, 3 or 4 substituents;

[0031] m is 0 or 1;

[0032] R4 is substituted or unsubstituted phenyl, substituted or unsubstituted thienyl, substituted or unsubstituted furyl;

[0033] The substituents in R4 may be selected from the following groups: F, Cl, Br, hydroxyl, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, vinyl, ethynyl, hydroxymethyl, carboxyl, sulfonic acid, boric acid, methylthio, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxycarbonyl, C1-C4 alkoxysulfonyl, C1-C4 alkylamino.

[0034] In another preferred embodiment, the compound has the structural formula as shown in (III):

[0035]

[0036] in,

[0037] R1 is hydrogen or methyl;

[0038] R2 is independently selected from hydrogen, F, Cl, Br, hydroxy, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, n represents 1, 2, 3 or 4 substituents;

[0039] R3 is hydrogen or methyl;

[0040] m is 0 or 1;

[0041] R4 is a substituted or unsubstituted phenyl group, a substituted or unsubstituted thienyl group;

[0042] The substituents in R4 may be selected from the following groups: F, Cl, Br, hydroxyl, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, vinyl, ethynyl, hydroxymethyl, carboxyl, sulfonic acid, boric acid, methylthio, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxycarbonyl, C1-C4 alkoxysulfonyl, C1-C4 alkylamino.

[0043] The pharmaceutically acceptable salts described in the present invention are salts formed by the compounds of the present invention and an acid selected from the group consisting of hydrochloric acid, p-toluenesulfonic acid, tartaric acid, maleic acid, lactic acid, methanesulfonic acid, sulfuric acid, phosphoric acid, citric acid, acetic acid or trifluoroacetic acid, preferably hydrochloric acid, p-toluenesulfonic acid or trifluoroacetic acid.

[0044] The compound according to any one of the first aspects of the present invention is the target compound of the present invention prepared in the examples (represented by structural formula or described by systematic nomenclature) and its isomers, and pharmaceutically acceptable salts thereof.

[0045] According to any one of the compounds of the first aspect of the present invention, it is a compound selected from the following:

[0046]

[0047]

[0048]

[0049]

[0050]

[0051]

[0052] The second aspect of the present invention provides a method for preparing any one of the compounds of the first aspect of the present invention, comprising the following steps:

[0053]

[0054] Compound A is reacted with compound B in a suitable solvent (e.g., dichloromethane, tetrahydrofuran, acetonitrile, 1,4-dioxane, preferably 1,4-dioxane) under the action of reagents CDMT and NMM, under the protection of air or inert gas (Ar or N2), at -10°C-50°C for 1-24 hours, preferably at room temperature for 2-3 hours, and then heated to reflux for 1-24 hours, preferably 6-12 hours, to obtain a compound of formula C; the compound of formula C is reacted in a suitable solvent (e.g., methanol, ethanol, methanol / water, ethanol / water, preferably methanol / water) under the action of ammonium chloride and metal powder (zinc powder, iron powder, preferably zinc powder) at -10°C-60°C for 1-24 hours, preferably at room temperature for 2-4 hours, to obtain a compound of formula D;

[0055]

[0056] The compound represented by formula D is reacted in a suitable solvent (such as DMF, DMSO, preferably DMF) with the compound R4Br under alkaline conditions (such as sodium carbonate, potassium carbonate, cesium carbonate, preferably potassium carbonate) under air or inert gas (Ar or N2) protection at -10°C-120°C for 1-24 hours, preferably 60°C-90°C for 4-8 hours to obtain the compound represented by formula (I); or the compound represented by D is reacted in a suitable solvent (such as tetrahydrofuran, acetonitrile, 1,4-dioxane, toluene, preferably toluene) with the compound R4Br under the action of reagents palladium acetate, BINAP and cesium carbonate, under inert gas (Ar or N2) protection at -10°C-120°C for 1-24 hours, preferably 90°C-110°C for 4-8 hours to obtain the compound represented by formula (I);

[0057]

[0058] The compound represented by formula D is reacted in a suitable solvent (e.g., dichloromethane, tetrahydrofuran, acetonitrile, 1,2-dichloroethane, preferably 1,2-dichloroethane) with compound R4CHO under acidic conditions (e.g., formic acid, acetic acid, preferably acetic acid) and a reducing agent (e.g., sodium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, preferably sodium triacetoxyborohydride) at -10°C-50°C for 1-24 hours, preferably at room temperature for 6-12 hours, to obtain a compound represented by formula (I);

[0059]

[0060] The compound represented by formula D is reacted with compound R3I in a suitable solvent (such as tetrahydrofuran, DMF, DMSO, acetone, preferably acetone) under alkaline conditions (such as sodium hydride, sodium carbonate, potassium carbonate, cesium carbonate, preferably potassium carbonate) under the protection of air or inert gas (Ar or N2) at -10°C-120°C for 1-24 hours, preferably at 0°C-70°C for 4-8 hours, to obtain a compound represented by formula E. The compound represented by formula E is reacted in a suitable solvent (e.g., dichloromethane, tetrahydrofuran, acetonitrile, 1,2-dichloroethane, preferably 1,2-dichloroethane) with the compound R4CHO under acidic conditions (e.g., formic acid, acetic acid, preferably acetic acid) and a reducing agent (e.g., sodium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, preferably sodium triacetoxyborohydride) at -10°C to 50°C for 1-24 hours, preferably at room temperature for 6-12 hours, to obtain the compound represented by formula (I);

[0061] The third aspect of the present invention provides use of the compound and its isomers, or pharmaceutically acceptable salts thereof, as described in any one of the first aspects of the present invention in the preparation of papain-like protease (PLpro) inhibitors.

[0062] The fourth aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the compound and its isomers according to any one of the first aspects of the present invention, or a pharmaceutically acceptable salt thereof, and optionally one or more pharmaceutically acceptable excipients.

[0063] The fifth aspect of the present invention provides the use of the compound and its isomers, or pharmaceutically acceptable salts thereof, described in any one of the first aspects of the present invention, or the pharmaceutical composition described in any one of the fourth aspects of the present invention in the preparation of drugs for treating and / or preventing infectious diseases caused by coronavirus.

[0064] The foregoing description only summarizes certain aspects of the present invention, but is not limited to these aspects. These aspects and other aspects will be described in more detail and complete below. DETAILED DESCRIPTION OF THE INVENTION

[0066] Various aspects and features of the present invention are further described below.

[0067] The various terms and phrases used in the present invention have the general meanings known to those skilled in the art. Even so, the present invention still hopes to provide a more detailed description and explanation of these terms and phrases. If the terms and phrases mentioned are inconsistent with the known meanings, the meanings expressed in the present invention shall prevail. The following are the definitions of various terms used in the present invention, which apply to the terms used in the entire specification of this application, unless otherwise specified in specific circumstances.

[0068] In general, the term "substituted or unsubstituted" means that one or more hydrogen atoms in a given structure are replaced by a specified substituent. Unless otherwise indicated, an optional substituent group can be substituted at each substitutable position of the group. When a given structure always has more than one position that can be substituted by one or more substituents selected from a specific group, the substituents can be substituted the same or differently at each position.

[0069] C i -C j It refers to a moiety having an integer "i" (inclusive) to an integer "j" (inclusive) number of carbon atoms. Thus, for example, C1-C3 alkyl refers to an alkyl group having 1 to 3 (inclusive) carbon atoms.

[0070] As used herein, the term "alkyl" refers to an alkyl group having a specified number of carbon atoms, which is a linear or branched alkyl group, and which may include its subgroups, for example, when referring to "C1-C3 alkyl", it may also include sub-range groups represented by C1-C2 alkyl, as well as specific groups such as methyl, ethyl, n-propyl, isopropyl.

[0071] As used herein, the terms "alkoxy" and "alkylamino" are conventional expressions and refer to an alkyl group attached to the remainder of the molecule through an oxygen atom or an amine group, respectively, wherein the alkyl group is as described herein.

[0072] As used herein, the term "haloalkyl" means that the hydrogen on the alkyl group is replaced by one or more halogen atoms, such examples include, but are not limited to, monofluoromethyl, monofluoromethoxy and the like.

[0073] As described herein, the term "cycloalkyl" refers to a cyclic alkyl group having a specified number of ring carbon atoms, and it may include its subgroups, for example, when referring to "C3-C7 cycloalkyl", it may also include sub-range groups represented by C3-C5 cycloalkyl, C4-C7 cycloalkyl, etc., as well as specific groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

[0074] As used herein, the term "ring" refers to a substituted or unsubstituted cycloalkyl group. The so-called ring includes a condensed ring. The number of atoms in the ring is usually defined as the number of ring members, for example, "C3-C6 ring" refers to 3-6 atoms arranged around.

[0075] As used herein, the terms "halogen," "halo," and the like refer to fluorine (F), chlorine (Cl), or bromine (Br).

[0076] In the present invention, "room temperature" refers to a temperature from 10°C to 40°C. In some embodiments, "room temperature" refers to a temperature from 20°C to 30°C; in other embodiments, room temperature refers to 25°C.

[0077] As used herein, the term "effective amount" refers to the amount of a drug that can achieve the desired treatment of a disease or condition described herein in a subject.

[0078] As used herein, the term "pharmaceutically acceptable" when describing, for example, a "pharmaceutically acceptable salt" means that the salt is not only physiologically acceptable to a subject, but also refers to a synthetic substance having pharmaceutical use value.

[0079] As described herein, the term "pharmaceutical composition", which may also refer to a "composition", can be used to achieve the treatment of the diseases or disorders described herein in a subject, especially a mammal.

[0080] "Treatment" for illness includes:

[0081] (1) preventing the disease, that is, preventing the clinical symptoms of the disease from occurring in a mammal that is exposed to or susceptible to the disease but does not experience or show symptoms of the disease,

[0082] (2) inhibiting the disease, i.e., arresting or reducing the progression of the disease or its clinical symptoms,

[0083] (3) Alleviate the disease, that is, cause recovery of the disease or its clinical symptoms.

[0084] "Therapeutically effective amount" refers to the amount of a compound that is sufficient to achieve treatment of a disease when administered to a mammal for the treatment of the disease. The therapeutically effective amount will vary depending on the compound, the disease to be treated and its severity, and the age, weight, sex, and other factors of the mammal. The therapeutically effective amount may also refer to any amount of the compound that is sufficient to achieve the desired beneficial effect, including preventing, inhibiting, or alleviating the disease as described in (1)-(3) above. For example, the amount of the compound may be between 0.1-250 mg / kg, or preferably, 0.5-100 mg / kg, or more preferably, 1-50 mg / kg, or even more preferably, 2-20 mg / kg. Preferably, the amount of the compound is administered to the mammal twice a day. More preferably, the amount of the compound is administered to the mammal once a day.

[0085] As described herein, the term "disease and / or condition" refers to a physical state of the subject, which is related to the disease and / or condition described in the present invention. For example, the disease and / or condition described in the present invention refers to a tuberculosis infectious disease.

[0086] As described herein, the term "subject" may refer to a patient or other animal, particularly a mammal, such as a human, monkey, etc., that receives the compound of formula I or its pharmaceutical composition for treating the disease or condition described in the present invention.

[0087] Another aspect of the present invention also relates to a pharmaceutical composition having the compound of the present invention as an active ingredient. The pharmaceutical composition can be prepared according to methods known in the art. The compound of the present invention can be combined with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants to prepare any dosage form suitable for human or animal use.

[0088] The compounds of the present invention or pharmaceutical compositions containing the same can be administered in unit dosage form, and the administration route can be enteral or parenteral, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eyes, lungs and respiratory tract, skin, vagina, rectum, etc.

[0089] The dosage form can be a liquid dosage form, a solid dosage form or a semisolid dosage form. Liquid dosage forms can be solutions (including true solutions and colloidal solutions), emulsions (including o / w types, w / o types and multiple emulsions), suspensions, injections (including water injections, powder injections and infusions), eye drops, nasal drops, lotions and liniments, etc.; solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules (including hard capsules, soft capsules, enteric-coated capsules), granules, powders, micropills, dropping pills, suppositories, films, patches, aerosols (powders), sprays, etc.; semisolid dosage forms can be ointments, gels, pastes, etc.

[0090] The compound of the present invention can be made into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

[0091] In order to prepare the compound of the present invention into tablets, various excipients known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, and solubilizers. The diluent may be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the wetting agent may be water, ethanol, isopropanol, etc.; the binder may be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinyl pyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and solubilizing agent may be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0092] The tablets can be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

[0093] In order to prepare the dosing unit into a capsule, the active ingredient compound of the present invention can be mixed with a diluent and a cosolvent, and the mixture can be directly placed in a hard capsule or a soft capsule. The active ingredient compound of the present invention can also be first prepared into granules or pellets with a diluent, a binder, and a disintegrant, and then placed in a hard capsule or a soft capsule. The diluents, binders, wetting agents, disintegrants, and cosolvents used to prepare tablets of the compound of the present invention can also be used to prepare capsules of the compound of the present invention.

[0094] To prepare the compound of the present invention into an injection, water, ethanol, isopropanol, propylene glycol or a mixture thereof can be used as a solvent and an appropriate amount of a solubilizer, cosolvent, pH adjuster, and osmotic pressure regulator commonly used in the art can be added. The solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; the pH adjuster can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. If a lyophilized powder injection is prepared, mannitol, glucose, etc. can also be added as a support agent.

[0095] Furthermore, if necessary, colorants, preservatives, perfumes, flavoring agents or other additives may be added to the pharmaceutical preparations.

[0096] In order to achieve the purpose of medication and enhance the therapeutic effect, the drug or pharmaceutical composition of the present invention can be administered by any known administration method.

[0097] The compound or composition of the present invention can be taken alone or in combination with other therapeutic drugs or symptomatic drugs. When the compound of the present invention has a synergistic effect with other therapeutic drugs, its dosage should be adjusted according to the actual situation.

[0098] Beneficial technical effects

[0099] The compounds of the present invention have strong PLpro inhibitory activity, and the inhibitory effects (IC 50 ) was less than 50 μM, and the PLpro inhibitory activity IC of 14 compounds was 50 <10μM. The antiviral activity of the three compounds was better than that of the PLpro inhibitor GRL0617, showing a strong anti-coronavirus effect. In addition, these compounds had low cytotoxicity to Vero E6 cells (IC 50 >50μM), indicating that the compounds have good safety. Metabolic stability tests of human and mouse liver microsomes show that the compounds of the present invention have good metabolic stability. The present invention provides a class of novel compounds with novel structures, strong in vitro PLpro inhibitory activity and low cytotoxicity, which can be used in drugs for inhibiting PLpro activity, especially as therapeutic agents for diseases related to coronavirus infection. DETAILED DESCRIPTION

[0100] The present invention can be described in detail by the following examples, but it is not intended to limit the present invention in any way. The present invention has been described in detail herein, and specific embodiments thereof are disclosed therein, and it is apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0101] For all of the following examples, standard manipulations and purification methods known to those skilled in the art may be used. The structures of the compounds were determined by H NMR ( 1 H NMR).

[0102] Preparation Examples Section

[0103] The structures of the compounds were determined by H NMR spectroscopy ( 1 H NMR). The H NMR shift (δ) is given in parts per million (ppm). The coupling constant (J) is in Hertz (Hz). The NMR spectrum is measured using a 400 Hz, 500 Hz or 700 Hz NMR instrument, deuterated dimethyl sulfoxide (DMSO-d6) or deuterated chloroform (CDCl3) as solvent, and tetramethylsilane (TMS) as internal standard.

[0104] Example 1

[0105] N-Cyclohexyl-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0106]

[0107] Synthesis route:

[0108]

[0109] Experimental steps:

[0110] The first step is the preparation of 3-(2-methyl-5-nitrophenyl)-5-(1-(naphthalen-2-yl)ethyl)-1,2,4-oxadiazole C-1

[0111] In a 100 mL reaction bottle, 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT, 878 mg, 5.0 mmol) and N-methylmorpholine (NMM, 1517 mg, 15.0 mmol) were dissolved in 1,4-dioxane (50 mL), stirred at room temperature for 15 minutes, 2-(naphthalene-2-yl)propionic acid (A-1, 1000 mg, 5.0 mmol) was added, stirred at room temperature for 30 minutes, (Z)-N'-hydroxy-2-methyl-5-nitrobenzenecarboxamidine (B-1, 976 mg, 5.0 mmol) was added, stirred at room temperature for 2 hours, and heated to reflux for 6 hours. Concentrated under reduced pressure, separated by silica gel (200-300 mesh) column chromatography, and ethyl acetate-petroleum ether (V: V = 0 ~ 5: 100) mixed solution was used as the eluent. Intermediate C-1 was obtained as a light yellow solid of 1.18 g, with a yield of 66%.

[0112] 1H NMR (400MHz, CDCl3) δ8.94(d,J=2.5Hz,1H),8.21(dd,J=8.5,2.5Hz,1H),8.16(d,J=8.3Hz,1H),7.90(d,J=8.8Hz,1H),7.82(d,J =8.9Hz,1H),7.61–7.56(m,1H),7.55–7.51(m,1H),7.51–7.45(m,3H),5.31(q,J=7.2Hz,1H),2.73(s,3H),2.00(d,J=7.2Hz,3H).

[0113] Step 2 Preparation of 4-methyl-3-(5-(1-(naphthalen-2-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline D-1

[0114] In a 25 mL reaction bottle, 3-(2-methyl-5-nitrophenyl)-5-(1-(naphthalen-2-yl)ethyl)-1,2,4-oxadiazole (C-1, 719 mg, 2.0 mmol) was dissolved in ethanol (10 mL), saturated aqueous ammonium chloride solution (2 mL) was added, stirred evenly, zinc powder (A-1, 650 mg, 10.0 mmol) was added, and stirred at room temperature for 3 hours. Filter, concentrate under reduced pressure, separate by silica gel (200-300 mesh) column chromatography, and use ethyl acetate-petroleum ether (V: V = 0 ~ 30: 100) mixed solution as eluent. Intermediate D-1 was obtained as an off-white solid 470 mg, with a yield of 71%.

[0115] 1 H NMR (500MHz, CDCl3) δ8.15(d,J=8.5Hz,1H),7.89(d,J=8.1Hz,1H),7.81(d,J=9.3Hz,1H),7.57(t,J=7.7Hz,1H),7.51(t,J=7.4Hz,1H),7. 47(d,J=7.0Hz,2H),7.38(s,1H),7.09(d,J=8.1Hz,1H),6.73(d,J=8.2Hz,1H),5.27(q,J=7.3Hz,1H),2.49(s,3H),1.97(d,J=7.2Hz,3H).

[0116] Step 3 Preparation of N-cyclohexyl-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline I-1

[0117] In a 25mL reaction bottle, 4-methyl-3-(5-(1-(naphthalene-2-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline (D-1, 165mg, 0.5mmol), bromocyclohexane (98mg, 0.6mmol), potassium carbonate (83mg, 0.6mmol) were dissolved in DMF (2mL), heated to 70℃ for 4 hours. Concentrated under reduced pressure, separated by silica gel (200-300 mesh) column chromatography, and ethyl acetate-petroleum ether (V:V=0~10:100) mixed solution was used as eluent. Compound I-1 (Compound 1) was obtained as an off-white solid of 138mg, with a yield of 67%.

[0118] 1 H NMR (400MHz, CDCl3) δ8.16(d,J=8.5Hz,1H),7.89(d,J=8.9Hz,1H),7.80(dd,J=7.4,2 .1Hz,1H),7.59–7.43(m,4H),7.25(d,J=2.8Hz,1H),7.07(d,J=8.2Hz,1H),6.65(dd,J =8.4,2.6Hz,1H),5.27(q,J=7.2Hz,1H),3.32–3.23(m,1H),2.46(s,3H),2.10–2.01(m ,2H),1.96(d,J=7.2Hz,3H),1.79–1.70(m,2H),1.40–1.30(m,2H),1.28–1.09(m,4H).

[0119] Example 2

[0120] N-(4,4-dimethylcyclohexyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0121]

[0122] Using 4-bromo-1,1-dimethylcyclohexane (115 mg, 0.6 mmol) as raw material, similar operation steps as the third step in Example 1 were adopted to react with intermediate D-1 to obtain compound I-2 (compound 2), 126 mg of off-white solid, with a yield of 57%.

[0123] 1H NMR (400MHz, CDCl3) δ8.16 (d, J=8.5Hz, 1H), 7.89 (d, J=8.7Hz, 1H), 7.80 (dd, J= 7.3,2.5Hz,1H),7.62–7.40(m,4H),7.28(s,1H),7.08(d,J=8.3Hz,1H),6.68(d ,J=8.2Hz,1H),5.27(q,J=7.2Hz,1H),3.28–3.18(m,1H),2.46(s,3H),1.96(d, J=7.2Hz,3H),1.93–1.84(m,2H),1.38–1.23(m,6H),0.94(s,3H),0.93(s,3H).

[0124] Example 3

[0125] N-(4-Cyanocyclohexyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0126]

[0127] Using 1-bromo-4-cyanocyclohexane (113 mg, 0.6 mmol) as raw material, similar operation steps as the third step in Example 1 were used to react with intermediate D-1 to obtain compound I-4 (compound 4), 155 mg of off-white solid, with a yield of 71%.

[0128] 1 H NMR (500MHz, CDCl3) δ8.16(d,J=8.5Hz,1H),7.89(d,J=8.1Hz,1H),7.81(d,J=7.6Hz,1 H),7.56(t,J=8.4Hz,1H),7.54–7.41(m,3H),7.25(s,1H),7.09(d,J=8.3Hz,1H),6.65( d,J=8.3Hz,1H),5.27(q,J=7.1Hz,1H),3.40–3.28(m,1H),2.89(t,J=4.3Hz,1H),2.47( s,3H),2.12–2.01(m,4H),1.97(d,J=7.2Hz,3H),1.76–1.66(m,2H),1.61–1.54(m,2H).

[0129] Example 4

[0130] N-(4-methoxycyclohexyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0131]

[0132] Using 1-bromo-4-methoxycyclohexane (116 mg, 0.6 mmol) as raw material, similar operation steps as the third step in Example 1 were adopted to react with intermediate D-1 to obtain compound I-4 (compound 4), 140 mg of off-white solid, and a yield of 63%.

[0133] 1 H NMR (400MHz, CDCl3) δ8.16(d,J=8.4Hz,1H),7.89(dd,J=7.9,1.9Hz,1H),7.80(dd,J=7.3,2.1Hz,1H),7.60–7.42(m,4H),7.30(s,1H),7.08(d,J=8.2H z,1H),6.70(s,1H),5.27(q,J=7.2Hz,1H),3.42–3.34(m,2H),3.31(s,3H) ,2.46(s,3H),1.96(d,J=7.2Hz,3H),1.89–1.75(m,4H),1.64–1.52(m,4H).

[0134] Example 5

[0135] 4-((4-methyl-3-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)cyclohexane-1-carboxylic acid

[0136]

[0137] Preparation of intermediate 4-methyl-3-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)aniline D-2

[0138]

[0139] Using 1-naphthylacetic acid as the starting material, similar operation steps as in the first and second steps of Example 1 were followed to obtain intermediate D-2.

[0140] 1 H NMR (400MHz, CDCl3) δ8.11(d,J=8.4Hz,1H),7.88(d,J=7.8Hz,1H),7.83(d,J=8.1Hz,1H),7.59–7.42(m, 4H),7.36(d,J=2.6Hz,1H),7.07(d,J=8.1Hz,1H),6.75(dd,J=8.1,2.6Hz,1H),4.72(s,2H),2.46(s,3H).

[0141] Preparation of 4-((4-methyl-3-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)cyclohexane-1-carboxylic acid I-5

[0142] Using 4-bromocyclohexane-1-carboxylic acid (124 mg, 0.6 mmol) as raw material, similar operation steps as the third step in Example 1 were used to react with intermediate D-2 to obtain compound I-5 (compound 5), 105 mg of off-white solid, with a yield of 48%.

[0143] 1 H NMR (400MHz, CDCl3) δ8.12(d,J=8.3Hz,1H),7.88(d,J=8.5Hz,1H),7.83(d,J=8.1 Hz,1H),7.59–7.42(m,4H),7.23(d,J=2.9Hz,1H),7.07(d,J=8.3Hz,1H),6.66(d, J=8.3Hz,1H),4.72(s,2H),3.31–3.21(m,1H),2.44(s,3H),2.32(q,J=4.8,4.2Hz ,1H),2.25–2.13(m,2H),2.13–2.04(m,2H),1.84–1.64(m,2H),1.64–1.50(m,2H).

[0144] Example 6

[0145] 4-((4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)cyclohexane-1-carboxylic acid

[0146]

[0147] Using 4-bromocyclohexane-1-carboxylic acid (124 mg, 0.6 mmol) as raw material, similar operation steps as the third step in Example 1 were used to react with intermediate D-1 to obtain compound I-6 (compound 6), 98 mg of off-white solid, with a yield of 43%.

[0148] 1H NMR (500MHz, CDCl3) δ8.15(d,J=8.5Hz,1H),7.88(d,J=8.1Hz,1H),7.80(d,J=7.6Hz,1H),7.56(t,J =7.6Hz,1H),7.53–7.42(m,3H),7.23(d,J=2.6Hz,1H),7.08(d,J=8.2Hz,1H),6.67–6.58(m,1H),5.2 7(q,J=7.2Hz,1H),3.28(t,J=11.0Hz,1H),2.46(s,3H),2.38–2.27(m,1H),2.20(d,J=12.0Hz,2H), 2.09(d,J=12.0Hz,2H), 1.96(d,J=7.2Hz,3H), 1.59(q,J=13.0Hz,2H), 1.14(q,J=13.4,12.6Hz,2H).

[0149] Example 7

[0150] 3-((4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)cyclohexane-1-carboxylic acid

[0151]

[0152] Using 3-bromocyclohexane-1-carboxylic acid (124 mg, 0.6 mmol) as raw material, similar operation steps as the third step in Example 1 were used to react with intermediate D-1 to obtain compound I-7 (compound 7), 135 mg of off-white solid, with a yield of 59%.

[0153] 1 H NMR (500MHz, CDCl3) δ8.14(d,J=8.5Hz,1H),7.88(d,J=8.0Hz,1H),7.80(d,J=7.7Hz,1 H),7.55(t,J=6.9Hz,1H),7.52–7.43(m,3H),7.30(s,1H),7.09(d,J=8.2Hz,1H),6.70 (d,J=8.2Hz,1H),5.26(q,J=7.2Hz,1H),3.69–3.62(m,1H),2.79–2.70(m,1H),2.47(s ,3H),2.22–2.11(m,1H),1.96(d,J=7.2Hz,3H),1.87–1.78(m,2H),1.77–1.55(m,5H).

[0154] Example 8

[0155] 3-((4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)cyclobutane-1-carboxylic acid

[0156]

[0157] Using 4-bromocyclobutane-1-carboxylic acid (107 mg, 0.6 mmol) as raw material, similar operation steps as the third step in Example 1 were used to react with intermediate D-1 to obtain compound I-8 (compound 8), 120 mg of off-white solid, with a yield of 56%.

[0158] 1 H NMR (400MHz, CDCl3) δ8.15(d,J=8.5Hz,1H),7.87(d,J=8.0Hz,1H),7.79(d,J=7.6Hz,1H),7.60–7.39(m,5H),7.10(d,J=8.2Hz,1H),6.63(d,J=5. 4Hz,1H),5.27(q,J=7.1Hz,1H),4.24–4.13(m,1H),3.22–3.12(m,1H),2 .79–2.69(m,2H),2.47(s,3H),2.25–2.14(m,2H),1.96(d,J=7.2Hz,3H).

[0159] Example 9

[0160]

[0161] Using 4-bromocyclobutane-1-carboxylic acid methyl ester (116 mg, 0.6 mmol) as raw material, similar operation steps as the third step in Example 1 were adopted to react with intermediate D-1 to obtain compound I-9 (compound 9), 113 mg of off-white solid, with a yield of 51%.

[0162] 1H NMR (400MHz, CDCl3) δ8.16(d,J=7.9Hz,1H),7.89(d,J=7.8Hz,1H),7.81(dd,J=7.0,2.5Hz,1H ),7.60–7.54(m,1H),7.54–7.50(m,1H),7.50–7.43(m,2H),7.24(d,J=2.6Hz,1H),7.09(d,J=8 .3Hz,1H),6.62(dd,J=8.3,2.6Hz,1H),5.27(q,J=7.2Hz,1H),4.00–3.85(m,1H),3.69(s,3H) ,2.95–2.80(m,1H),2.78–2.63(m,2H),2.47(s,3H),2.18–2.06(m,2H),1.97(d,J=7.2Hz,3H).

[0163] Example 10

[0164] 3-((4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)cyclopentane-1-carboxylic acid methyl ester

[0165]

[0166] Using 3-bromocyclopentane-1-carboxylic acid methyl ester (124 mg, 0.6 mmol) as raw material, similar operation steps as the third step in Example 1 were used to react with intermediate D-1 to obtain compound I-10 (compound 10), 110 mg of off-white solid, with a yield of 48%.

[0167] 1 H NMR (400MHz, CDCl3) δ8.16(d,J=8.5Hz,1H),7.89(d,J=9.8Hz,1H),7.80(dd,J=7.2,2.3Hz,1H) ,7.59–7.54(m,1H),7.54–7.43(m,3H),7.29(d,J=2.6Hz,1H),7.10(dd,J=8.3,3.9Hz,1H),6.69 (dd,J=8.2,2.7Hz,1H),5.27(q,J=7.2Hz,1H),4.01–3.89(m,1H),3.68(s,3H),2.93–2.82(m,1H ),2.47(s,3H),2.38–2.05(m,3H),2.02–1.98(m,2H),1.97(d,J=7.3Hz,3H),1.89–1.78(m,1H).

[0168] Embodiment 11

[0169] 6-((4-methyl-3-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)spiro[3.3]heptane-2-carboxylic acid

[0170]

[0171] Using 6-bromospiro[3.3]heptane-2-carboxylic acid (131 mg, 0.6 mmol) as raw material, similar operation steps as in the third step of Example 1 were used to react with intermediate D-2 to obtain compound I-11 (compound 11), 100 mg of off-white solid, with a yield of 44%.

[0172] 1 H NMR (400MHz, CDCl3) δ8.13(d,J=8.3Hz,1H),7.88(d,J=7.7Hz,1H),7.84(d,J=8.1H z,1H),7.59–7.43(m,4H),7.15(s,1H),7.07(d,J=8.2Hz,1H),6.59(dd,J=8.3,2.6H z,1H),4.73(s,2H),3.86–3.72(m,1H),3.10–3.01(m,1H),2.61–2.54(m,1H),2.51– 2.46(m,1H),2.44(s,3H),2.42–2.28(m,3H),2.26–2.16(m,1H),1.93–1.78(m,2H).

[0173] Example 12

[0174] 6-((4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)spiro[3.3]heptane-2-carboxylic acid

[0175]

[0176] Using 6-bromospiro[3.3]heptane-2-carboxylic acid (131 mg, 0.6 mmol) as raw material, similar operation steps as in the third step of Example 1 were used to react with intermediate D-1 to obtain compound I-12 (compound 12), 110 mg of off-white solid, with a yield of 47%.

[0177] 1H NMR (400MHz, CDCl3) δ8.15(d,J=8.4Hz,1H),7.89(d,J=9.6Hz,1H),7.80(d,J=9.6Hz,1H),7. 62–7.40(m,4H),7.22(d,J=2.6Hz,1H),7.09(d,J=8.3Hz,1H),6.61(dd,J=8.2,2.6Hz,1H),5. 31–5.22(m,1H),3.86–3.76(m,1H),3.12–3.00(m,1H),2.61–2.54(m,1H),2.51–2.48(m,1H), 2.47(s,3H),2.40–2.29(m,3H),2.25–2.17(m,1H),1.96(d,J=7.2Hz,3H),1.94–1.80(m,2H).

[0178] Embodiment 13

[0179] N-Benzyl-4-methyl-3-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)aniline

[0180]

[0181] Experimental steps:

[0182]

[0183] In a 25 mL reaction bottle, 4-methyl-3-(5-(naphthalene-1-ylmethyl)-1,2,4-oxadiazol-3-yl)aniline (D-2, 126 mg, 0.4 mmol), benzaldehyde (85 mg, 0.8 mmol), glacial acetic acid (144 mg, 2.4 mmol) were dissolved in 1,2-dichloroethane (10 mL), sodium triacetoxyborohydride (506 mg, 2.4 mmol) was added, and stirred at room temperature for 3 hours. Concentrated under reduced pressure, separated by 200-300 mesh silica gel column chromatography, and ethyl acetate-petroleum ether (V: V = 0 ~ 10: 100) mixed solution was used as eluent. Compound I-13 (Compound 13) was obtained, 121 mg of off-white solid, and the yield was 75%.

[0184] 1H NMR (400MHz, CDCl3) δ8.12(d,J=8.3Hz,1H),7.91–7.86(m,1H),7.83(d,J=8.2Hz,1H),7.58–7.40(m,5H),7. 38–7.25(m,5H),7.06(d,J=8.2Hz,1H),6.64(dd,J=8.2,2.7Hz,1H),4.71(s,2H),4.33(s,2H),2.44(s,3H).

[0185] Embodiment 14

[0186] N-(4-bromobenzyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0187]

[0188] Using 4-bromobenzaldehyde (148 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-1 to obtain compound I-14 (Compound 14), 140 mg of off-white solid, with a yield of 70%.

[0189] 1 H NMR (400MHz, CDCl3) δ8.14(d,J=8.4Hz,1H),7.89(d,J=8.8Hz,1H),7.80(dd,J=6.2,3.3Hz,1H),7.59–7.48(m,2H),7.48–7.40(m,4H),7.33(d,J=2.6Hz ,1H),7.23(d,J=8.2Hz,2H),7.08(d,J=8.2Hz,1H),6.63(dd,J=8.2,2.6Hz, 1H), 5.26 (q, J = 7.2Hz, 1H), 4.30 (s, 2H), 2.47 (s, 3H), 1.95 (d, J = 7.2Hz, 3H).

[0190] Embodiment 15

[0191] N-(4-cyanobenzyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0192]

[0193] Using 4-cyanobenzaldehyde (105 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-1 to obtain compound I-15 (compound 15), 124 mg of off-white solid, with a yield of 70%.

[0194] 1 H NMR (400MHz, CDCl3) δ8.13(d,J=8.4Hz,1H),7.89(d,J=8.0Hz,1H),7.84–7.78(m,1H),7.58(d,J=8.4Hz,2H),7.56–7.50(m,2H),7.47–7.42(m,4H) ,7.28(d,J=2.7Hz,1H),7.07(d,J=8.3Hz,1H),6.58(dd,J=8.3,2.7Hz,1H ),5.25(q,J=7.2Hz,1H),4.41(s,2H),2.47(s,3H),1.95(d,J=7.2Hz,3H).

[0195] Example 16

[0196] N-(4-Hydroxybenzyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0197]

[0198] Using 4-hydroxybenzaldehyde (98 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-1 to obtain compound I-16 (compound 16), 130 mg of off-white solid, with a yield of 75%.

[0199] 1 H NMR (400MHz, CDCl3) δ8.12(d,J=8.4Hz,1H),7.87(d,J=6.0Hz,1H),7.78(d,J=7.8Hz,1H),7.56–7.40(m,4H),7.27(d,J=2.6Hz,1H),7.13(d,J=8.4Hz, 2H),7.07(d,J=8.2Hz,1H),6.71(d,J=8.5Hz,2H),6.63(dd,J=8.2,2.6Hz,1 H),5.27(q,J=7.2Hz,1H),4.14(s,2H),2.47(s,3H),1.95(d,J=7.2Hz,3H).

[0200] Embodiment 17

[0201] N-(Benzo[d][1,3]dioxolan-5-ylmethyl)-4-methyl-3-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)aniline

[0202]

[0203] Using benzo[d][1,3]dioxolane-5-carboxaldehyde (120 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-2 to obtain compound I-17 (compound 17), 121 mg of off-white solid, with a yield of 67%.

[0204] 1 H NMR (400MHz, CDCl3) δ8.11(d,J=8.3Hz,1H),7.88(d,J=7.9Hz,1H),7.83(d,J=8.1Hz,1H),7.59–7.41(m,4H),7.27(s,1H),7.0 6(d,J=8.3Hz,1H),6.83(s,1H),6.82–6.69(m,2H),6.63(d,J=5.4Hz,1H),5.92(s,2H),4.71(s,2H),4.22(s,2H),2.44(s,3H).

[0205] Embodiment 18

[0206] N-(4-methoxycarbonylbenzyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0207]

[0208] Using methyl p-formylbenzoate (131 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-1 to obtain compound I-18 (compound 18), 160 mg of off-white solid, with a yield of 84%.

[0209] 1H NMR (400MHz, CDCl3) δ8.14(d,J=8.5Hz,1H),8.01–7.95(m,2H),7.89(d,J=7.6Hz,1H),7.84–7.76(m,1H),7.60–7.29(m,7H),7.0 8(d,J=8.2Hz,1H),6.65(d,J=11.1Hz,1H),5.26(q,J=7.3Hz,1H),4.42(s,2H),3.90(s,3H),2.47(s,3H),1.95(d,J=7.2Hz,3H).

[0210] Embodiment 19

[0211] N-(4-carboxybenzyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0212]

[0213] Using p-formylbenzoic acid (120 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were adopted to react with intermediate D-1 to obtain compound I-19 (compound 19), 143 mg of off-white solid, and the yield was 77%.

[0214] 1 H NMR (400MHz, CDCl3) δ8.14(d,J=8.4Hz,1H),8.05(d,J=8.4Hz,2H),7.88(d,J=7.7Hz,1H),7.83–7.77(m,1H),7.59–7.49(m,2H),7.49–7.43 (m,4H),7.39(s,1H),7.09(d,J=8.2Hz,1H),6.68(d,J=8.3Hz,1H),5.26(q,J=7.1Hz,1H),4.45(s,2H),2.48(s,3H),1.96(d,J=7.2Hz,3H).

[0215] Embodiment 20

[0216] N-(4-Methanesulfonylbenzyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0217]

[0218] Using p-formylbenzene methanesulfonic acid (149 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were adopted to react with intermediate D-1 to obtain compound I-20 (compound 20), 162 mg of off-white solid, with a yield of 81%.

[0219] 1 H NMR (400MHz, CDCl3) δ8.14(d,J=8.4Hz,1H),7.93–7.83(m,3H),7.83–7.76(m,1H),7.59–7.49(m,4H),7.46(d,J=5.7Hz,2H),7.29(d,J=2.7Hz ,1H),7.08(d,J=8.2Hz,1H),6.59(dd,J=8.3,2.7Hz,1H),5.26(q,J=7.2Hz,1H),4.45(s,2H),3.01(s,3H),2.47(s,3H),1.95(d,J=7.2Hz,3H).

[0220] Embodiment 21

[0221] N-(3-Carboxybenzyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0222]

[0223] Using m-formylbenzoic acid (120 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-1 to obtain compound I-21 (Compound 21), 125 mg of off-white solid, with a yield of 67%.

[0224] 1 H NMR (400MHz, CDCl3) δ8.14(d,J=8.4Hz,1H),8.10(s,1H),7.99(d,J=7.7Hz,1H) ,7.87(d,J=7.2Hz,1H),7.79(dd,J=6.9,2.5Hz,1H),7.60(d,J=7.7Hz,1H),7.57 –7.38(m,5H),7.35(d,J=2.6Hz,1H),7.08(d,J=8.3Hz,1H),6.64(dd,J=8.2,2.6 Hz,1H),5.26(q,J=7.2Hz,1H),4.41(s,2H),2.47(s,3H),1.95(d,J=7.3Hz,3H).

[0225] Embodiment 22

[0226] N-(4-carboxybenzyl)-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0227]

[0228] Preparation of intermediate 2-(5-(1-(naphthalen-2-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline D-3

[0229]

[0230] Using (Z)-N'-hydroxy-3-nitrobenzamide as the starting material, similar operation steps as the first and second steps in Example 1 were used to obtain intermediate D-3.

[0231] 1 H NMR (400MHz, CDCl3) δ8.12(d,J=8.4Hz,1H),7.88(d,J=8.1Hz,1H),7.80(s,1H),7.60–7.38(m ,6H),7.28–7.21(m,1H),6.80(d,J=7.5Hz,1H),5.25(q,J=7.6Hz,1H),1.95(d,J=7.3Hz,3H).

[0232] Preparation of N-(4-carboxybenzyl)-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0233] Using p-formylbenzoic acid (120 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were adopted to react with intermediate D-3 to obtain compound I-22 (compound 22), 133 mg of off-white solid, with a yield of 74%.

[0234] 1 H NMR (400MHz, CDCl3) δ8.11(d,J=8.4Hz,1H),8.04(d,J=7.8Hz,2H),7.86(d,J=8.0Hz,1H),7.81–7.74(m,1H),7.53(t,J=7.5Hz,1H),7.49(d,J=7.6 Hz,1H),7.47–7.39(m,5H),7.36(s,1H),7.23(t,J=8.0Hz,1H),6.67(d,J =10.5Hz,1H),5.24(q,J=7.2Hz,1H),4.42(s,2H),1.93(d,J=7.2Hz,3H).

[0235] Embodiment 23

[0236] N-(4-carboxybenzyl)-3-trifluoromethyl-5-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0237]

[0238] Preparation of intermediate 3-trifluoromethyl-5-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)aniline D-4

[0239]

[0240] Using (Z)-N'-hydroxy-3-trifluoromethyl-5-nitrobenzenecarboxamidine as the starting material, similar operation steps as the first and second steps in Example 1 were used to obtain intermediate D-4.

[0241] 1 H NMR (400MHz, CDCl3) δ8.11(d,J=8.4Hz,1H),7.89(d,J=8.1Hz,1H),7.83–7.78(m,1H),7.74(s,1H) ,7.61–7.48(m,3H),7.47–7.44(m,2H),7.01(s,1H),5.26(q,J=6.7Hz,1H),1.96(d,J=7.2Hz,3H).

[0242] Preparation of N-(4-carboxybenzyl)-3-trifluoromethyl-5-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0243] Using p-formylbenzoic acid (120 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were adopted to react with intermediate D-4 to obtain compound I-23 (compound 23), 162 mg of off-white solid, with a yield of 78%.

[0244] 1 H NMR (400MHz, CDCl3) δ8.10(d,J=8.5Hz,1H),8.06(d,J=7.8Hz,2H),7.87(d,J=8.0Hz,1H),7.84–7.75(m,1H),7.69(s,1H),7.55(t ,J=7.6Hz,1H),7.50(d,J=7.7Hz,1H),7.48–7.39(m,5H),6.90(s,1H),5.26(q,J=7.1Hz,1H),4.45(s,2H),1.95(d,J=7.1Hz,3H).

[0245] Embodiment 24

[0246] 5-(((3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)furan-2-carboxylic acid

[0247]

[0248] Using 5-formyl-2-furancarboxylic acid (112 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-3 to obtain compound I-24 (Compound 24), 123 mg of off-white solid, with a yield of 70%.

[0249] 1 H NMR (400MHz, CDCl3) δ8.12(d,J=8.4Hz,1H),7.90(d,J=8.0Hz,1H),7.85–7.77(m,1H),7.61–7.48(m,2H),7.48–7.42(m,3H),7.42–7.37( m,1H),7.24(t,J=8.0Hz,1H),6.87(s,1H),6.80(d,J=8.1Hz,1H),6.28(s,1H),5.28(d,J=7.3Hz,1H),4.35(s,2H),1.96(d,J=7.2Hz,3H).

[0250] Embodiment 25

[0251] 5-(((4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)furan-2-carboxylic acid

[0252]

[0253] Using 5-formyl-2-furancarboxylic acid (112 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-1 to obtain compound I-25 (Compound 25), 137 mg of off-white solid, with a yield of 76%.

[0254] 1H NMR (400MHz, CDCl3) δ8.13(d,J=5.4Hz,1H),7.88(d,J=5.4Hz,1H),7.84–7.74(m,1H),7.60–7.48(m,2H),7.48–7.43(m,2H),7.36(d,J=3.0Hz,1H) ,7.04(dd,J=8.4,3.0Hz,1H),6.83(s,1H),6.67(d,J=8.5Hz,1H),6.20(s ,1H),5.31–5.20(m,1H),4.26(s,2H),2.44(s,3H),1.95(d,J=3.7Hz,3H).

[0255] Embodiment 26

[0256] 5-(((3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0257]

[0258] Using 5-formyl-2-thiophenecarboxylic acid (125 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-3 to obtain compound I-26 (compound 26), 105 mg of off-white solid, with a yield of 58%.

[0259] 1 H NMR (400MHz, CDCl3) δ8.12(d,J=8.3Hz,1H),7.88(d,J=8.0Hz,1H),7.80(t,J=5.1Hz,1H),7.75–7.67(m,1H),7.63–7.36( m,6H),7.26(d,J=7.4Hz,1H),7.03(s,1H),6.77(d,J=8.1Hz,1H),5.32–5.21(m,1H),4.60(s,2H),1.95(d,J=7.2Hz,3H).

[0260] Embodiment 27

[0261] 5-(((4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0262]

[0263] Using 5-formyl-2-thiophenecarboxylic acid (125 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-1 to obtain compound I-27 (Compound 27), 121 mg of off-white solid, with a yield of 64%.

[0264] 1 H NMR (400MHz, CDCl3) δ8.15(d,J=8.4Hz,1H),7.89(d,J=8.9Hz,1H),7.80(dd,J=6.7,2.9 Hz,1H),7.72(d,J=3.8Hz,1H),7.56(t,J=6.8Hz,1H),7.52(d,J=7.0Hz,1H),7.50–7.45( m,2H),7.37(d,J=2.7Hz,1H),7.11(d,J=8.3Hz,1H),7.02(d,J=3.8Hz,1H),6.70(dd,J= 8.2, 2.7Hz, 1H), 5.27 (q, J = 7.1Hz, 1H), 4.57 (s, 2H), 2.49 (s, 3H), 1.96 (d, J = 7.3Hz, 3H).

[0265] Embodiment 28

[0266] 5-(((2-methyl-5-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0267]

[0268] Preparation of intermediate 2-methyl-5-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)aniline D-5

[0269]

[0270] Using (Z)-N'-hydroxy-4-methyl-3-nitrobenzenecarboxamidine as the starting material, similar operation steps as the first and second steps in Example 1 were used to obtain intermediate D-5.

[0271] 1H NMR (400MHz, CDCl3) δ8.13(d,J=8.5Hz,1H),7.88(d,J=8.0Hz,1H),7.82–7.77(m,1H),7.60–7.47(m,2H),7.4 7–7.43(m,3H),7.42(s,1H),7.14(d,J=7.7Hz,1H),5.25(q,J=7.2Hz,1H),2.22(s,3H),1.95(d,J=7.2Hz,3H).

[0272] Preparation of 5-(((2-methyl-5-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid I-28

[0273] Using 5-formyl-2-thiophenecarboxylic acid (125 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-5 to obtain compound I-28 (compound 28), 110 mg of off-white solid, with a yield of 59%.

[0274] 1 H NMR (400MHz, CDCl3) δ8.13(d,J=8.3Hz,1H),7.88(d,J=8.2Hz,1H),7.84–7.76(m,1H),7.73(d,J=4.0Hz,1H),7.60–7.42(m,5 H),7.36(s,1H),7.18(d,J=7.7Hz,1H),7.07(s,1H),5.26(q,J=7.1Hz,1H),4.67(s,2H),2.23(s,3H),1.95(d,J=7.2Hz,3H).

[0275] Embodiment 29

[0276] 5-(((3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)-5-(trifluoromethyl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0277]

[0278] Using 5-formyl-2-thiophenecarboxylic acid (125 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-4 to obtain compound I-29 (compound 29), 132 mg of off-white solid, and a yield of 63%.

[0279] 1H NMR (400MHz, CDCl3) δ8.11(d,J=8.5Hz,1H),7.89(d,J=8.0Hz,1H),7.81(t,J=5.0Hz,1H),7.74(s,2H), 7.62–7.43(m,5H),7.05(s,1H),6.96(s,1H),5.27(q,J=7.2Hz,1H),4.63(s,2H),1.96(d,J=7.1Hz,3H).

[0280] Embodiment 30

[0281] 5-(((2-methoxy-5-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0282]

[0283] Preparation of intermediate 2-methoxy-5-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)aniline D-6

[0284]

[0285] Using (Z)-N'-hydroxy-4-methoxy-3-nitrobenzamide as the starting material, similar operation steps as the first and second steps in Example 1 were used to obtain intermediate D-6.

[0286] 1 H NMR(400MHz, CDCl3)δ8.13(d,J=8.3Hz,1H),7.88(d,J=7.9Hz,1H),7.84–7.77(m,1H),7.61–7 .42(m,6H),6.84(d,J=8.4Hz,1H),5.24(q,J=7.3Hz,1H),3.90(s,3H),1.94(d,J=7.2Hz,3H).

[0287] Preparation of 5-(((2-methoxy-5-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0288] Using 5-formyl-2-thiophenecarboxylic acid (125 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-6 to obtain compound I-30 (compound 30), 120 mg of off-white solid, with a yield of 62%.

[0289] 1H NMR (400MHz, CDCl3) δ8.13(d,J=8.4Hz,1H),7.88(d,J=8.0Hz,1H),7.82–7.76(m,1H),7.72(d,J=3.6Hz,1H),7.59–7.47(m,3H),7.47–7.43 (m,2H),7.36(s,1H),7.07(d,J=3.7Hz,1H),6.85(d,J=8.3Hz,1H),5.24(q,J=7.1Hz,1H),4.65(s,2H),3.91(s,3H),1.94(d,J=7.2Hz,3H).

[0290] Embodiment 31

[0291] N-((5-chlorothien-2-yl)methyl)-3-(5-(1-(naphthalen-1-yl)ethyl)-4-methyl-1,2,4-oxadiazol-3-yl)aniline

[0292]

[0293] Using 5-chlorothiophene-2-carboxaldehyde (117 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-1 to obtain compound I-31 (compound 31), 110 mg of off-white solid, with a yield of 60%.

[0294] 1 H NMR (400MHz, CDCl3) δ8.15(d,J=8.5Hz,1H),7.89(d,J=8.0Hz,1H),7.81(d,J=7.0Hz,1H),7.61–7.43(m,4H),7.38(s, 1H),7.12(d,J=8.3Hz,1H),6.79–6.68(m,3H),5.27(q,J=7.3Hz,1H),4.44(s,2H),2.49(s,3H),1.96(d,J=7.2Hz,3H).

[0295] Embodiment 32

[0296] N-((5-bromothiophen-2-yl)methyl)-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)-5-(trifluoromethyl)aniline

[0297]

[0298] Using 5-bromothiophene-2-carboxaldehyde (153 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-4 to obtain compound I-32 (compound 32), 150 mg of off-white solid, with a yield of 67%.

[0299] 1 H NMR (400MHz, CDCl3) δ8.12(d,J=8.5Hz,1H),7.90(d,J=8.0Hz,1H),7.84–7.78(m,1H),7.73(s,1H),7.57(t,J=7.7Hz,1H),7.58–7 .54(m,2H),7.48–7.43(m,2H),6.96(s,1H),6.90(s,1H),6.79(s,1H),5.27(q,J=7.8Hz,1H),4.51(s,2H),1.96(d,J=5.0Hz,3H).

[0300] Embodiment 33

[0301] N-((5-Nitrothiophen-2-yl)methyl)-3-(5-(1-(naphthalen-1-yl)ethyl)-4-methyl-1,2,4-oxadiazol-3-yl)aniline

[0302]

[0303] Using 5-nitrothiophene-2-carboxaldehyde (126 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-1 to obtain compound I-33 (compound 33), 130 mg of yellow solid, with a yield of 69%.

[0304] 1 H NMR(400MHz, CDCl3)δ8.13(d,J=8.4Hz,1H),7.89(d,J=6.9Hz,1H),7.85–7.73(m,2H),7.60–7.40(m,4H),7.32(s,1H),7.1 1(d,J=8.3Hz,1H),6.92(s,1H),6.66(d,J=8.3Hz,1H),5.31–5.22(m,1H),4.54(s,2H),2.49(s,3H),1.96(d,J=7.2Hz,3H).

[0305] Embodiment 34

[0306] N-((5-methoxythiophen-2-yl)methyl)-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)-5-(trifluoromethyl)aniline

[0307]

[0308] Using 5-methoxythiophene-2-carboxaldehyde (114 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-4 to obtain compound I-34 (compound 34), 144 mg of off-white solid, with a yield of 71%.

[0309] 1 H NMR(400MHz, CDCl3) δ8.12(d,J=8.3Hz,1H),7.90(d,J=8.0Hz,1H),7.86–7.76 (m,1H),7.75–7.69(m,1H),7.57(t,J=7.6Hz,1H),7.55–7.48(m,2H),7.48–7.4 4(m,2H),6.96(s,1H),6.66–6.55(m,1H),6.04–5.98(m,1H),5.27(q,J=7.2Hz ,1H),4.59(s,1H),4.41(s,1H),3.84(d,J=10.5Hz,3H),1.96(d,J=7.2Hz,3H).

[0310] Embodiment 35

[0311] N-((5-methoxycarbonylthiophen-2-yl)methyl)-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)-5-(trifluoromethyl)aniline

[0312]

[0313] Using 5-formylthiophene-2-carboxylic acid methyl ester (136 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-4 to obtain compound I-35 (compound 35), 113 mg of off-white solid, with a yield of 53%.

[0314] 1H NMR (400MHz, CDCl3) δ8.19–8.04(m,1H),7.90(t,J=6.7Hz,1H),7.82(s,1H),7.74(d,J=5.3Hz,1H),7.67(t,J=4. 3Hz,1H),7.63–7.39(m,5H),7.08–6.85(m,2H),5.36–5.16(m,1H),4.61(s,2H),3.87(s,3H),2.01–1.90(m,3H).

[0315] Embodiment 36

[0316] N-((5-methoxycarbonylthiophen-2-yl)methyl)-5-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)-2-methylaniline

[0317]

[0318] Using 5-formylthiophene-2-carboxylic acid methyl ester (136 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-5 to obtain compound I-36 (compound 36), 130 mg of off-white solid, and a yield of 67%.

[0319] 1 H NMR (400MHz, CDCl3) δ8.13(d,J=8.4Hz,1H),7.88(d,J=8.0Hz,1H),7.79(s,1H),7.66(s,1H),7.60–7.40(m,5H),7.34(s,1H) ),7.16(d,J=7.7Hz,1H),7.02(s,1H),5.24(d,J=7.1Hz,1H),4.63(s,2H),3.84(s,3H),2.21(s,3H),1.94(d,J=7.2Hz,3H).

[0320] Embodiment 37

[0321] 5-(((3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-boronic acid

[0322]

[0323] Using 5-formyl-2-thiopheneboronic acid (125 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were adopted to react with intermediate D-3 to obtain compound I-37 (compound 37), 110 mg of off-white solid, with a yield of 60%.

[0324] 1 H NMR(400MHz, CDCl3)δ8.13(d,J=8.4Hz,1H),7.88(d,J=8.1Hz,1H),7.85–7.76(m,1H),7.60–7.43(m,4H),7.40(s,1H),7.31–7.24(m,1H) ,7.21(d,J=5.0Hz,1H),7.02(s,1H),6.95(t,J=4.4Hz,1H),6.78(d,J=8.2Hz,1H),5.29–5.20(m,1H),4.56(s,2H),1.95(d,J=7.2Hz,3H).

[0325] Embodiment 38

[0326] N-(4-(Methylsulfonyl)benzyl)-4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0327]

[0328] Preparation of intermediate 4-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)aniline D-7

[0329]

[0330] Using (Z)-N'-hydroxy-4-nitrobenzamide as the starting material, similar operation steps as the first and second steps in Example 1 were used to obtain intermediate D-7.

[0331] 1 H NMR (400MHz, DMSO-d6) δ8.20(d,J=8.3Hz,1H),7.99(d,J=6.3Hz,1H),7.90(d,J=8.1Hz,1H),7.65(d,J=8.4Hz,2H),7.64–7.54(m,2H ),7.51(t,J=7.7Hz,1H),7.42(d,J=7.1Hz,1H),6.64(d,J=8.6Hz,2H),5.75(brs,2H),5.41(q,J=7.0Hz,1H),1.84(d,J=7.1Hz,3H).

[0332] Preparation of N-(4-(methylsulfonyl)benzyl)-4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0333] Using 4-methylsulfonylbenzaldehyde (147 mg, 0.8 mmol) as raw material, adopting the similar operation steps in the third step of Example 13, reacting with intermediate D-7 to obtain compound I-38 (compound 38), 130 mg of off-white solid, yield 67%.

[0334] 1 H NMR (400MHz, CDCl3) δ8.12(d,J=8.4Hz,1H),7.94–7.86(m,5H),7.82–7.76(m,1H),7.56(d,J=7.6Hz,2H),7.54–7.48(m,2 H),7.44(d,J=4.8Hz,2H),6.72(d,J=8.2Hz,2H),5.23(q,J=7.2Hz,1H),4.52(s,2H),3.04(s,3H),1.93(d,J=7.2Hz,3H).

[0335] Embodiment 39

[0336] 4-(((4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)benzoic acid

[0337]

[0338] Using p-formylbenzoic acid (120 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were adopted to react with intermediate D-7 to obtain compound I-39 (compound 39), 111 mg of off-white solid, with a yield of 62%.

[0339] 1 H NMR (400MHz, DMSO-d6) δ8.19(d,J=8.4Hz,1H),7.98(d,J=7.9Hz,1H),7.93–7. 87(m,3H),7.69(d,J=8.4Hz,2H),7.63–7.53(m,2H),7.50(t,J=7.7Hz,1H),7. 46(d,J=8.0Hz,2H),7.41(d,J=7.2Hz,1H),7.02(t,J=6.2Hz,1H),6.67(d,J=8 .5Hz,2H),5.40(q,J=7.0Hz,1H),4.43(d,J=6.0Hz,2H),1.83(d,J=7.0Hz,3H).

[0340] Embodiment 40

[0341] 5-(((4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-boronic acid

[0342]

[0343] Using 5-formyl-2-thiopheneboronic acid (125 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-7 to obtain compound I-40 (compound 40), 102 mg of off-white solid, with a yield of 56%.

[0344] 1 H NMR (700MHz, CDCl3) δ8.13(d,J=8.5Hz,1H),7.96–7.87(m,2H),7.79(d,J=7.1Hz,1H),7.61–7.41(m,4H),7.23(s ,1H),7.02(s,1H),6.96(s,1H),6.70(d,J=8.3Hz,2H),5.23(d,J=7.1Hz,1H),4.56(s,2H),1.94(d,J=7.5Hz,3H).

[0345] Embodiment 41

[0346] N-((5-Nitrothiophen-2-yl)methyl)-4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0347]

[0348] Using 5-nitrothiophene-2-carboxaldehyde (126 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-7 to obtain compound I-41 (compound 41), a yellow solid 106 mg, with a yield of 57%.

[0349] 1 H NMR (400MHz, CDCl3) δ8.13(d,J=8.4Hz,1H),7.92(d,J=8.3Hz,2H),7.88(d,J=8.0Hz,1H),7.81–7.78(m,2H),7.59–7.47(m,2H ),7.47–7.42(m,2H),6.97(d,J=4.1Hz,1H),6.70(d,J=8.2Hz,2H),5.24(q,J=7.1Hz,1H),4.60(s,2H),1.94(d,J=7.5Hz,3H).

[0350] Embodiment 42

[0351] N-((5-methoxycarbonylthiophen-2-yl)methyl)-4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0352]

[0353] Using 5-formylthiophene-2-carboxylic acid methyl ester (136 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-7 to obtain compound I-42 (compound 42), 133 mg of off-white solid, with a yield of 71%.

[0354] 1 H NMR (400MHz, CDCl3) δ8.13(d,J=8.4Hz,1H),7.95–7.84(m,3H),7.82–7.76(m,1H),7.66(d,J=3.8Hz,1H),7.59–7.47(m,2H),7.47– 7.40(m,2H),6.99(d,J=3.7Hz,1H),6.68(d,J=8.3Hz,2H),5.23(q,J=7.2Hz,1H),4.58(s,2H),3.85(s,3H),1.94(d,J=7.2Hz,3H).

[0355] Embodiment 43

[0356] 5-(((4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0357]

[0358] Using 5-formyl-2-thiophenecarboxylic acid (125 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-7 to obtain compound I-43 (compound 43), 110 mg of off-white solid, with a yield of 60%.

[0359] 1H NMR (400MHz, CDCl3) δ8.13(d,J=8.5Hz,1H),7.95–7.85(m,3H),7.82–7.75(m,1H),7.73(d,J=4.3Hz,1H),7.59–7.47(m,2H ),7.47–7.42(m,2H),7.02(d,J=3.8Hz,1H),6.79–6.65(m,2H),5.23(q,J=7.3Hz,1H),4.60(s,2H),1.94(d,J=7.2Hz,3H).

[0360] Embodiment 44

[0361] 5-(((2-methyl-4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0362]

[0363] Preparation of intermediate 2-methyl-4-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)aniline D-8

[0364]

[0365] Using (Z)-N'-hydroxy-3-methyl-4-nitrobenzamide as the starting material, similar operation steps as the first and second steps in Example 1 were used to obtain intermediate D-8.

[0366] 1 H NMR (400MHz, CDCl3) δ8.13(d,J=8.4Hz,1H),7.88(d,J=7.5Hz,1H),7.82–7.75(m,3H),7.59–7.47(m,2H ),7.47–7.43(m,2H),6.74(d,J=8.2Hz,1H),5.24(q,J=7.2Hz,1H),2.22(s,3H),1.94(d,J=7.2Hz,3H).

[0367] Preparation of 5-(((2-methyl-4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0368] Using 5-formyl-2-thiophenecarboxylic acid (125 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-8 to obtain compound I-44 (compound 44), 116 mg of off-white solid, and the yield was 62%.

[0369] 1 H NMR (700MHz, CDCl3) δ8.13(d,J=8.5Hz,1H),7.88(d,J=6.8Hz,1H),7.85–7.81( m,2H),7.79(dd,J=6.3,3.1Hz,1H),7.76(d,J=3.8Hz,1H),7.55(t,J=6.9Hz,1H) ,7.50(t,J=6.9Hz,1H),7.47–7.43(m,2H),7.05(d,J=3.8Hz,1H),6.65(d,J=8.3 Hz,1H),5.24(q,J=7.2Hz,1H),4.67(s,2H),2.23(s,3H),1.94(d,J=7.2Hz,3H).

[0370] Embodiment 45

[0371] 5-(((2-Fluoro-4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0372]

[0373] Preparation of intermediate 2-fluoro-4-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)aniline D-9

[0374]

[0375] Using (Z)-N'-hydroxy-3-fluoro-4-nitrobenzamide as the starting material, similar operation steps as the first and second steps in Example 1 were used to obtain intermediate D-9.

[0376] 1 H NMR(400MHz, CDCl3)δ8.13(d,J=8.4Hz,1H),7.89(d,J=8.0Hz,1H),7.84–7.78(m,1H),7.77–7.69(m,2H), 7.60–7.48(m,2H),7.48–7.43(m,2H),6.87(t,J=8.5Hz,1H),5.24(q,J=7.2Hz,1H),1.94(d,J=7.2Hz,3H).

[0377] Preparation of 5-(((2-fluoro-4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0378] Using 5-formyl-2-thiophenecarboxylic acid (125 mg, 0.8 mmol) as raw material, similar operation steps as the third step in Example 13 were used to react with intermediate D-9 to obtain compound I-45 (compound 45), 130 mg of off-white solid, with a yield of 69%.

[0379] 1 H NMR (700MHz, CDCl3) δ8.12(d,J=8.5Hz,1H),7.88(d,J=8.1Hz,1H),7.80(t,J=4.8Hz,1H),7.78–7.72(m,2H),7.56(t,J=7.7Hz,1H),7.50(t,J=7.5H z,1H),7.45(d,J=4.8Hz,2H),7.26(s,1H),7.05(d,J=3.8Hz,1H),6.72(t ,J=8.4Hz,1H),5.24(q,J=7.2Hz,1H),4.65(s,2H),1.94(d,J=7.1Hz,3H).

[0380] Embodiment 46

[0381] 5-((Methyl(4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)methyl)thiophene-2-carboxylic acid

[0382]

[0383] Preparation of Intermediate N-methyl-4-(5-(naphthalen-1-ylmethyl)-1,2,4-oxadiazol-3-yl)-aniline D-10

[0384]

[0385] In a 10 mL reaction bottle, 4-(5-(naphthalene-1-ylmethyl)-1,2,4-oxadiazol-3-yl)-aniline D-9 (315 mg, 1.0 mmol) was dissolved in 5 mL of acetone, potassium carbonate (138 mg, 1.0 mmol) and iodomethane (142 mg, 1.0 mmol) were added, and heated to reflux for 4 hours. The mixture was concentrated under reduced pressure, and separated by silica gel (200-300 mesh) column chromatography, with a methanol-dichloromethane (V:V=0-5:100) mixture as the eluent. Intermediate D-10 was obtained as an off-white solid (227 mg), with a yield of 69%.

[0386] 1H NMR (400MHz, CDCl3) δ8.14(d,J=8.4Hz,1H),7.92(d,J=8.6Hz,2H),7.88(d,J=8.1Hz,1H),7.79(d,J=6.5Hz,1H),7.59–7.53(m,1H), 7.51(d,J=7.6Hz,1H),7.49–7.40(m,2H),6.66(d,J=8.7Hz,2H),5.24(q,J=7.2Hz,1H),2.88(d,J=1.8Hz,3H),1.94(d,J=7.2Hz,3H).

[0387] Using 5-formyl-2-thiophenecarboxylic acid (126 mg, 0.8 mmol) and N-methyl-(4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline as raw materials (132 mg, 0.4 mmol), similar operation steps as the third step in Example 13 were adopted to react with intermediate D-10 to obtain compound I-46 (compound 46), 117 mg of off-white solid, and a yield of 62%.

[0388] 1 H NMR (400MHz, CDCl3) δ8.13(d,J=8.4Hz,1H),7.96(d,J=8.6Hz,2H),7.88(d,J=8. 0Hz,1H),7.79(dd,J=6.6,2.9Hz,1H),7.71(d,J=3.8Hz,1H),7.55(t,J=7.4Hz,1H ),7.51(d,J=7.6Hz,1H),7.49–7.42(m,2H),6.94(d,J=3.8Hz,1H),6.83(d,J=8. 6Hz, 2H), 5.24 (q, J = 7.2Hz, 1H), 4.75 (s, 2H), 3.11 (s, 3H), 1.94 (d, J = 7.2Hz, 3H).

[0389] Embodiment 47

[0390] N-(4-methoxycarbonylphenyl)-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0391]

[0392] route:

[0393]

[0394] Experimental steps:

[0395] In a 10 mL reaction bottle, 3-(5-(1-(naphthalene-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline (D-2, 156 mg, 0.5 mmol), methyl p-bromobenzoate (129 mg, 0.6 mmol), palladium acetate (7 mg, 0.03 mmol), BINAP (25 mg, 0.04 mmol), cesium carbonate (227 mg, 0.07 mmol) were added with 5 mL of anhydrous toluene and stirred evenly, and heated to reflux for 8 hours under argon protection. Concentrated under reduced pressure, separated by 200-300 mesh silica gel column chromatography, ethyl acetate-petroleum ether (V: V = 0 ~ 10: 100) mixed solution as eluent, to obtain compound I-47 (compound 47), off-white solid 144 mg, yield 64%.

[0396] 1 H NMR (400MHz, CDCl3) δ8.13 (d, J=8.3Hz, 1H), 7.97–7.73 (m, 6H), 7.60–7.39 (m, 5H), 7.30 (d, J= 8.2Hz, 1H), 7.02 (d, J = 8.3Hz, 2H), 5.27 (q, J = 7.1Hz, 1H), 3.88 (s, 3H), 1.96 (d, J = 7.2Hz, 3H).

[0397] Embodiment 48

[0398] N-(4-methoxycarbonylphenyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0399]

[0400] Using methyl p-bromobenzoate (129 mg, 0.6 mmol) as raw material, similar operation steps as in Example 47 were used to react with intermediate D-1 to obtain compound I-48 (compound 48), 134 mg of off-white solid, with a yield of 58%.

[0401] 1 H NMR (400MHz, CDCl3) δ8.14(d,J=8.4Hz,1H),7.94–7.86(m,3H),7.83–7.77(m,2H),7.56(t,J=7.6Hz,1H),7.52(d,J=7.5Hz,1H),7. 50–7.43(m,2H),7.27–7.19(m,2H),6.94(d,J=8.7Hz,2H),5.27(q,J=7.1Hz,1H),3.87(s,3H),2.58(s,3H),1.96(d,J=7.2Hz,3H).

[0402] Embodiment 49

[0403] N-(4-methoxycarbonylphenyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)methyl)-1,2,4-oxadiazol-3-yl)aniline

[0404]

[0405] Using methyl p-bromobenzoate (129 mg, 0.6 mmol) as raw material, similar operation steps as in Example 47 were used to react with intermediate D-2 to obtain compound I-49 (compound 49), 141 mg of off-white solid, with a yield of 63%.

[0406] 1 H NMR (400MHz, CDCl3) δ8.12(d,J=8.1Hz,1H),7.93–7.87(m,3H),7.84(d,J=8.1Hz,1H),7.76(s,1H),7.59–7.50(m,3H),7.4 6(t,J=7.6Hz,1H),7.24(d,J=7.5Hz,1H),7.21–7.15(m,1H),6.93(d,J=8.5Hz,2H),4.72(s,2H),3.88(s,3H),2.55(s,3H).

[0407] Embodiment 50

[0408] N-(3-methoxycarbonylphenyl)-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0409]

[0410] Using methyl m-bromobenzoate (129 mg, 0.6 mmol) as raw material, adopt similar operation steps as Example 47 to react with intermediate D-3 to obtain compound I-50 (Compound 50), 122 mg of off-white solid, and the yield is 52%.

[0411] 1H NMR (400MHz, CDCl3) δ8.13(d,J=8.5Hz,1H),7.89(d,J=8.0Hz,1H),7.83–7.78(m,1H),7.74(d,J=11.9Hz,2H),7.68(d,J=7.8H z,1H),7.61(d,J=7.4Hz,1H),7.59–7.42(m,4H),7.41–7.19(m,4H),5.26(q,J=7.1Hz,1H),3.89(s,3H),1.96(d,J=7.2Hz,3H).

[0412] Embodiment 51

[0413] N-(4-Carboxyphenyl)-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0414]

[0415] Using p-bromobenzoic acid (121 mg, 0.6 mmol) as raw material, similar operation steps as in Example 47 were adopted to react with intermediate D-3 to obtain compound I-51 (compound 51), 112 mg of off-white solid, with a yield of 51%.

[0416] 1 H NMR (400MHz, DMSO-d6) δ8.98(brs,1H),8.22(d,J=8.4Hz,1H),8.00(d,J=9.7Hz,1H),7.91(d,J=8.1Hz,1H),7.83(d,J=8 .7Hz,2H),7.78(t,J=1.9Hz,1H),7.67–7.38(m,8H),7.12(d,J=8.7Hz,2H),5.50(q,J=7.1Hz,1H),1.88(d,J=7.1Hz,3H).

[0417] Embodiment 52

[0418] N-(4-Carboxyphenyl)-4-methyl-3-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0419]

[0420] Using p-bromobenzoic acid (121 mg, 0.6 mmol) as raw material, similar operation steps as in Example 47 were adopted to react with intermediate D-1 to obtain compound I-52 (compound 52), 132 mg of off-white solid, with a yield of 59%.

[0421] 1 H NMR (400MHz, CDCl3) δ8.15(d,J=8.5Hz,1H),7.97(d,J=8.7Hz,2H),7.89(d,J=8.0Hz,1H),7.85–7.78(m,2H),7.60 –7.43(m,4H),7.31–7.21(m,2H),6.96(d,J=8.8Hz,2H),5.28(q,J=7.2Hz,1H),2.59(s,3H),1.97(d,J=7.2Hz,3H).

[0422] Embodiment 53

[0423] N-(4-Carboxyphenyl)-4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0424]

[0425] Using p-bromobenzoic acid (121 mg, 0.6 mmol) as raw material, similar operation steps as in Example 47 were adopted to react with intermediate D-7 to obtain compound I-53 (compound 53), 150 mg of off-white solid, with a yield of 69%.

[0426] 1 H NMR (400MHz, DMSO-d6) δ9.13(brs,1H),8.21(d,J=7.8Hz,1H),7.98(d,J=8.0Hz,1H),7.96–7.81(m,5H),7.67–7.47( m,3H),7.47–7.38(m,1H),7.30(d,J=5.7Hz,2H),7.20(d,J=6.0Hz,2H),5.45(q,J=6.7Hz,1H),1.86(d,J=4.4Hz,3H).

[0427] Embodiment 54

[0428] N-(3-Carboxyphenyl)-4-(5-(1-(naphthalen-1-yl)ethyl)-1,2,4-oxadiazol-3-yl)aniline

[0429]

[0430] Using m-bromobenzoic acid (121 mg, 0.6 mmol) as raw material, similar operation steps as in Example 47 were adopted to react with intermediate D-7 to obtain compound I-54 (compound 54), 127 mg of off-white solid, with a yield of 58%.

[0431] 1 H NMR (400MHz, DMSO-d6) δ8.69(brs,1H),8.22(d,J=8.3Hz,1H),8.00(d,J=7.8Hz,1H),7.91(d,J=8. 1Hz,1H),7.71(d,J=10.8Hz,2H),7.66–7.29(m,10H),5.50(q,J=7.1Hz,1H),1.88(d,J=7.0Hz,3H).

[0432] Embodiment 55

[0433] N-(3-methoxycarbonylphenyl)-4-(5-(1-(naphthalen-1-yl)methyl)-1,2,4-oxadiazol-3-yl)aniline

[0434]

[0435] Using methyl m-bromobenzoate (129 mg, 0.6 mmol) as raw material, adopt similar operation steps as Example 47 to react with intermediate D-7 to obtain compound I-55 (compound 55), 150 mg of off-white solid, and the yield is 67%.

[0436] 1 H NMR (400MHz, CDCl3) δ8.13(d,J=8.4Hz,1H),7.89(d,J=8.0Hz,1H),7.80(s,1H),7.74(d,J=11.9Hz,2H),7.68(d,J=7.7Hz,1H),7. 61(d,J=7.3Hz,1H),7.59–7.48(m,2H),7.46(s,2H),7.41–7.18(m,4H),5.26(q,J=7.7Hz,1H),3.89(s,3H),1.96(d,J=7.2Hz,3H).

[0437] Embodiment 56

[0438] N-(4-methoxycarbonylphenyl)-4-(5-(1-(naphthalen-1-yl)methyl)-1,2,4-oxadiazol-3-yl)aniline

[0439]

[0440] Using methyl p-bromobenzoate (129 mg, 0.6 mmol) as raw material, adopt similar operation steps as Example 47 to react with intermediate D-7 to obtain compound I-56 (compound 56), 166 mg of off-white solid, with a yield of 74%.

[0441] 1H NMR (400MHz, CDCl3) δ8.14(d,J=8.6Hz,1H),8.03(d,J=8.0Hz,2H),7.96(d,J=7.7Hz,2H),7.89(d,J=8.2Hz,1H),7.80(d,J=6.9Hz,1H),7.60 –7.50(m,2H),7.46(d,J=7.1Hz,2H),7.20(d,J=7.8Hz,2H),7.09(d,J=7.8Hz,2H),5.26(q,J=7.5Hz,1H),3.89(s,3H),1.96(d,J=6.8Hz,3H).

[0442] Embodiment 57

[0443] N-(4-cyanophenyl)-4-(5-(1-(naphthalen-1-yl)methyl)-1,2,4-oxadiazol-3-yl)aniline

[0444]

[0445] Using 4-bromobenzonitrile (109 mg, 0.6 mmol) as raw material, similar operation steps as in Example 47 were adopted to react with intermediate D-7 to obtain compound I-57 (compound 57), 133 mg of off-white solid, with a yield of 62%.

[0446] 1 H NMR (400MHz, DMSO-d6) δ9.28(brs,1H),8.22(d,J=8.3Hz,1H),8.00(d,J=8.0Hz,1H),7.97–7.88(m,3H),7.67(d,J=8.2Hz,2H),7. 65–7.49(m,3H),7.44(d,J=7.4Hz,1H),7.32(d,J=8.2Hz,2H),7.22(d,J=8.4Hz,2H),5.47(q,J=7.4Hz,1H),1.87(d,J=7.0Hz,3H).

[0447] Embodiment 58

[0448] N-(3-cyanophenyl)-4-(5-(1-(naphthalen-1-yl)methyl)-1,2,4-oxadiazol-3-yl)aniline

[0449]

[0450] Using 3-bromobenzonitrile (109 mg, 0.6 mmol) as raw material, similar operation steps as in Example 47 were adopted to react with intermediate D-7 to obtain compound I-58 (compound 58), 140 mg of off-white solid, with a yield of 67%.

[0451] 1 H NMR (400MHz, CDCl3) δ8.14(d,J=8.6Hz,1H),8.02(d,J=8.3Hz,2H),7.89(d,J=8.2Hz,1H),7.81(s,1H),7.63–7. 41(m,4H),7.41–7.19(m,3H),7.12(d,J=8.3Hz,2H),6.06(s,1H),5.26(q,J=7.5Hz,1H),1.96(d,J=7.0Hz,3H).

[0452] Biological activity test

[0453] 1. In vitro protease inhibition activity test

[0454] Determination method: Fluorescence method was used to determine the half maximal inhibitory concentration (IC) of the compound against SARS-CoV-2 PLpro protease. 50 ).

[0455] Experimental principle: The characteristic substrate of PLpro protease is labeled with fluorescence quenching. If the substrate is digested, it will emit characteristic fluorescence, and if it is not digested, it will not emit fluorescence. The digestion rate of the substrate by the protease is detected by measuring the fluorescence value at different inhibitor concentrations, and the half-inhibitory concentration value (IC50) of the compound against PLpro is obtained according to the digestion reaction rate at different inhibitor concentrations. 50 ).

[0456] Experimental methods:

[0457] (1) Use DMSO to dilute the compound in series.

[0458] (2) Add reaction buffer, test compounds of different concentrations and PLpro protease in a 96-well transparent-bottom black plate (Greiner, 655096) according to the reaction system.

[0459] (3) Place the reaction plate in a 37°C incubator for 8 to 10 minutes to allow the enzymatic reaction to proceed, then add the fluorescent substrate.

[0460] (4) Use an enzyme-labeled instrument to detect fluorescence intensity in real time.

[0461] (5) The data obtained from the test were fitted using Origin and Graphpad to obtain the IC 50 Numeric value.

[0462] Experimental Results

[0463] Table 1. Inhibitory effects of the compounds of the present invention on PLpro

[0464]

[0465]

[0466] From the data in Table 1, it can be seen that the compounds of the present invention have good PLpro inhibitory effect. The IC values ​​of the 14 compounds for PLpro inhibitory activity are 50 <10 μM.

[0467] 2. Cytotoxicity assay

[0468] Determination method: MTT method Experimental principle: Cell activity is determined by the reduction of oxidized 3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyl tetrazolium bromide (trade name: Thiazole Blue) / MTT [3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyl tetrazolium bromide] to an insoluble blue formazan compound through mitochondrial dehydrogenase (such as succinate dehydrogenase), which is then dissolved in DMSO and then colored for determination. The conversion amount is positively correlated with the number of living cells.

[0469] Experimental methods:

[0470] (1) Digest VeroE6 cells that have been cultured to the logarithmic growth phase with 0.25% trypsin for 2 to 3 minutes, discard the digestion solution, add an appropriate amount of culture medium, mix well, take 20 μL, count under a microscope using a hemocytometer, and prepare a cell suspension of appropriate concentration for later use. At the same time, use PBS (phosphate buffered solution) to prepare

[0471] 5g / L MTT solution was filtered and sterilized for later use.

[0472] (2) The test drug was dissolved in DMSO and diluted 50-fold with culture medium to make the highest test concentration. The test drug was then serially diluted with culture medium at a ratio of 1:2 on a 96-well plate. Eight concentrations were set for each compound, with the highest concentration being 200 μM. Four parallel wells were set for each concentration, with 100 μL / well.

[0473] (3) The prepared cell suspension was inoculated into a 96-well plate at 100 μL / well with a cell concentration of 4×10 5cell / mL. At the same time, set up cell control wells without drug and culture medium blank control wells.

[0474] (4) After 48 hours of culture, add 10 μL / well of MTT and continue culturing for 4 hours. Remove the culture plate, carefully discard the culture medium in the wells, add 100 μL of DMSO to each well, shake until the formazan particles are completely dissolved, and measure the optical density (OD) at a wavelength of 570 nm using an enzyme-linked immunosorbent assay. 570 ).

[0475] (5) Data processing: Cell survival percentage (%) = [(cell control OD 570 Value - OD of drug-added group 570 value) / (cell control OD 570 Value - Blank OD 570 The dose-response curve was fitted using Origin software to calculate the concentration of each compound at which 50% of the cells survived (CC 50 ).

[0476] Experimental results:

[0477] Table 2. Cytotoxicity results of the compounds of the present invention

[0478]

[0479] It can be seen from the data in Table 2 that the compounds of the present invention have low cytotoxicity and exhibit high safety.

[0480] 3. Antiviral activity detection

[0481] Determination method: Fluorescence quantitative PCR method to detect the total amount of RNA in the supernatant of cell culture medium Experimental principle: During PCR amplification, a specific fluorescent probe is added at the same time as a pair of primers. The probe is an oligonucleotide with a reporter fluorescent group and a quencher fluorescent group labeled at both ends. When the probe is intact, the fluorescent signal emitted by the reporter group is absorbed by the quencher group; at the beginning, the probe binds to any single strand of DNA. During PCR amplification, the 5'-3' exonuclease activity of the Taq enzyme will enzymatically degrade the probe, separating the reporter fluorescent group and the quencher fluorescent group, so that the fluorescent monitoring system can receive the fluorescent signal, that is, for each DNA chain amplified, a fluorescent molecule is formed, achieving complete synchronization between the accumulation of the fluorescent signal and the formation of the PCR product.

[0482] Experimental methods:

[0483] (1) VeroE6 cells that have been cultured to the logarithmic growth phase are digested with 0.25% trypsin for 2 to 3 minutes, the digestion solution is discarded, an appropriate amount of culture medium is added, mixed, and 20 μL is taken out for counting under a microscope using a hemacytometer to prepare a cell suspension of appropriate concentration for later use.

[0484] (2) The test drug was dissolved in DMSO and diluted 50-fold with culture medium to make the highest test concentration. The test drug was then serially diluted with culture medium at a ratio of 1:2 on a 96-well plate. Eight concentrations were set for each compound, with the highest concentration being 200 μM. Four parallel wells were set for each concentration, with 100 μL / well.

[0485] (3) The prepared cell suspension was inoculated into a 96-well plate at 100 μL / well with a cell concentration of 4×10 5 cell / mL.

[0486] (4) After incubating cells with different dilutions of drug working solution (200ul) for 1h, 100TCID 50 The 2019-Wuhan strain virus was incubated for 2 hours.

[0487] (5) Remove the cell culture medium from the 96-well culture plate and add different concentrations of compound dilutions, 200 μl per well, and 3 replicates per concentration. At the same time, set up drug-free cell control wells, culture medium blank control wells, solvent control wells, and virus control wells.

[0488] (6) After culturing for 48 hours, the culture supernatant was added to AVL lysis buffer and repeatedly pipetted for 15 seconds to ensure sufficient lysis. RNA was then extracted from the cell culture supernatant according to the RNA extraction kit procedure.

[0489] (7) After the extracted RNA, fluorescent probe and polymerase are evenly mixed, the RNA in the supernatant is quantitatively detected using the QPCR method.

[0490] (8) Data processing: Cell antiviral activity = [(Ct value of drug-treated group - Ct value of virus control group / (Ct value of cell control group - Ct value of virus control group)] × 100%. GraphPad software was used to fit the dose-response curve and calculate the concentration of each compound at which the antiviral activity reached 50% (EC 50 ).

[0491] Experimental results:

[0492] Table 3. Antiviral test results of compounds

[0493]

[0494] It can be seen from the data in Table 3 that the antiviral activities of compounds 25, 43 and 46 of the present invention are better than that of the PLpro inhibitor GRL0617, showing a strong anti-coronavirus effect.

[0495] 4. Liver microsome metabolic stability test

[0496] Experimental method: The target compounds were selected for metabolic stability study in liver microsomes (mixed human origin, Bioreclamation). The specific method is as follows: The synthesized target compounds were prepared into 1 μM test solutions.

[0497] The microsomal protein concentration was 1 mg / mL. The reaction was initiated by adding NADPH (1 mM), and the samples were incubated in a shaking incubator at 37°C for up to 60 minutes. The reaction was terminated at 0, 5, 15 and 30 minutes by adding ice-cold acetonitrile / methanol (50:50) containing an internal standard. Aliquots of the reaction mixture were taken at 0, 5, 15, 30 and 60 minutes, respectively, and then ice-cold acetonitrile / methanol (50:50, v / v) containing an internal standard was added. The samples were centrifuged at 4°C for 15 minutes (4000 rpm), and the supernatant was analyzed by LC-MS / MS. Chromatographic conditions: Column: Kinetex C18 100A (30 mm × 3.0 mm, 2.6 μm); Column temperature: room temperature, Mobile phase: acetonitrile-water (containing 0.1% formic acid) gradient; Flow rate: 0.9 mL / min. The metabolic stability of the compounds was assessed by measuring the residual amount of the compounds in the presence or absence of NADPH cofactor.

[0498] Table 4 Metabolic stability data of human and mouse liver microsomes of some compounds of the present invention

[0499]

[0500] It can be seen from the data in Table 4 that compounds 29 and 43 of the present invention show good metabolic stability in human and mouse liver microsomes.

Claims

1. A compound represented by formula (I) and its isomers, or a pharmaceutically acceptable salt thereof: in, R1 is hydrogen, methyl, ethyl, propyl, isopropyl; R2 is independently selected from hydrogen, F, Cl, Br, hydroxy, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, n represents 1, 2, 3 or 4 substituents; R3 is hydrogen, methyl, ethyl, propyl; m is 0 or 1; R4 is substituted or unsubstituted C3-C7 cycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted thienyl, substituted or unsubstituted furanyl, substituted or unsubstituted spiro[3,3]heptyl; The substituents in R4 may be selected from the following groups: F, Cl, Br, hydroxyl, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, vinyl, ethynyl, hydroxymethyl, carboxyl, sulfonic acid, methanesulfonic acid, boric acid, methylthio, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxycarbonyl, C1-C4 alkoxysulfonyl, C1-C4 alkylamino.

2. A compound represented by formula (II) and its isomers, or a pharmaceutically acceptable salt thereof: in, R1 is hydrogen or methyl; R2 is independently selected from hydrogen, F, Cl, Br, hydroxy, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, n represents 1, 2, 3 or 4 substituents; m is 0 or 1; R4 is substituted or unsubstituted C3-C7 cycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted thienyl, substituted or unsubstituted furanyl, substituted or unsubstituted spiro[3,3]heptyl; The substituents in R4 may be selected from the following groups: F, Cl, Br, hydroxyl, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, vinyl, ethynyl, hydroxymethyl, carboxyl, sulfonic acid, methanesulfonic acid, boric acid, methylthio, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxycarbonyl, C1-C4 alkoxysulfonyl, C1-C4 alkylamino.

3. The compound according to claim 2, its isomer, or its pharmaceutically acceptable salt: in, R1 is hydrogen or methyl; R2 is independently selected from hydrogen, F, Cl, Br, hydroxy, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, n represents 1, 2, 3 or 4 substituents; m is 0; R4 is substituted or unsubstituted C3-C7 cycloalkyl, substituted or unsubstituted spiro[3,3]heptyl; The substituents in R4 may be selected from the following groups: F, Cl, Br, hydroxyl, amino, nitro, cyano, hydroxymethyl, carboxyl, sulfonic acid, methanesulfonic acid, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxycarbonyl.

4. The compound according to claim 2, its isomer, or its pharmaceutically acceptable salt: in, R1 is hydrogen or methyl; R2 is independently selected from hydrogen, F, Cl, Br, hydroxy, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, n represents 1, 2, 3 or 4 substituents; m is 0 or 1; R4 is substituted or unsubstituted phenyl, substituted or unsubstituted thienyl, substituted or unsubstituted furyl; The substituents in R4 may be selected from the following groups: F, Cl, Br, hydroxyl, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, vinyl, ethynyl, hydroxymethyl, carboxyl, sulfonic acid, methanesulfonic acid, boric acid, methylthio, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxycarbonyl, C1-C4 alkoxysulfonyl, C1-C4 alkylamino.

5. A compound represented by formula (III) and its isomers, or a pharmaceutically acceptable salt thereof: in, R1 is hydrogen or methyl; R2 is independently selected from hydrogen, F, Cl, Br, hydroxy, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, n represents 1, 2, 3 or 4 substituents; R3 is hydrogen or methyl; m is 0 or 1; R4 is a substituted or unsubstituted phenyl group, a substituted or unsubstituted thienyl group; The substituents in R4 may be selected from the following groups: F, Cl, Br, hydroxyl, amino, nitro, cyano, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, vinyl, ethynyl, hydroxymethyl, carboxyl, sulfonic acid, methanesulfonic acid, boric acid, methylthio, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxycarbonyl, C1-C4 alkoxysulfonyl, C1-C4 alkylamino.

6. The compound according to any one of claims 1 to 5 and a pharmaceutically acceptable salt thereof, which is selected from the following compounds:

7. A method for preparing a compound according to any one of claims 1 to 6, comprising the following steps: Compound A and compound B undergo condensation and ring closure to obtain compound C, which is then reduced to obtain intermediate D; when m=0 and R4 is a cycloalkyl group, D and R4Br undergo a substitution reaction under alkaline conditions to obtain a compound of formula (I); when m=0 and R4 is an aryl group, D and R4Br undergo a CN coupling reaction to obtain a compound of formula (I); compound D and R4CHO undergo an amination reduction to obtain a compound of formula (I); compound D and R3I undergo a substitution reaction to obtain compound E, which is then subjected to an amination reduction with R4CHO to obtain a compound of formula (I); Wherein, R1, R2, R3, R4, m, and n are defined as any one of claims 1-5.

8. Use of the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof in the preparation of a papain-like protease (PLpro) inhibitor.

9. A pharmaceutical composition, characterized in that The composition comprises a therapeutically and / or prophylactically effective amount of the compound according to any one of claims 1 to 6 and a pharmaceutically acceptable salt thereof and optionally one or more pharmaceutically acceptable carriers, excipients, diluents, adjuvants and vehicles.

10. Use of the compound according to any one of claims 1 to 6 and a pharmaceutically acceptable salt thereof or the composition according to claim 9 in the preparation of a medicament for treating and / or preventing infectious diseases caused by coronavirus.

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