Regulator of sphingosine 1 phosphate receptor subtype 1 and application thereof

Abstract

The invention discloses a modifier of sphingosine 1 phosphate receptor subtype 1 and application of the modifier, and belongs to the field of medicines. The regulator prepared by the invention is a compound shown as a formula I-S or I-R, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester, prodrug, homologue, hydrate or solvate thereof. The invention provides a sphingosine-1-phosphate receptor subtype 1 regulator with polyfluoro-substituted alkyl, the agonistic activity of the sphingosine-1-phosphate receptor subtype 1 of the compound is high, and EC50 is in the range of 10 <-9 >-10 <-10 > M. The regulator prepared by the preparation method disclosed by the invention has good S1PR1 agonist activity.

Description

Technical Field

[0001] The invention belongs to the field of medicines and relates to a regulator of sphingosine 1-phosphate receptor subtype 1 and application thereof. Background Art

[0002] The sphingosine 1-phosphate receptor subtype 1 (S1P1 / EDG1 receptor) is a G-protein coupled receptor (GPCR) and a member of the endothelial differentiation gene (EDG) receptor family. Endogenous ligands for the EDG receptor include lysophospholipids, such as sphingosine 1-phosphate (S1P). Like all GPCRs, ligation of the receptor transmits second messenger signals through the activation of G-proteins (α, β and γ).

[0003] The development of small molecule S1P1 receptor agonists and antagonists has provided insight into the physiological role of the S1P1 / S1P-receptor signaling system. Activation of the S1P1 receptor interferes with lymphocyte trafficking, sequestering them in lymph nodes and other secondary lymphoid tissues, which leads to rapid and irreversible lymphopenia, and this may be due to receptor engagement of lymphatic endothelial cells and lymphocytes themselves (Rosen et al., Immunol. Rev., 2003, 195: 160-177). The clinically valuable result of lymphocyte sequestration is to reduce lymphocytes in surrounding inflammatory tissues and inhibit inflammatory responses.

[0004] Although fingolimod, siponimod, bonecimod and ozanimod are currently on the market for the treatment of multiple sclerosis, these compounds still have the risk of bradycardia. Even fingolimod, siponimod and bonecimod require ECG monitoring when taking the medicine for the first time. All four drugs require a dose-escalation regimen to reduce the incidence of bradycardia. Therefore, more effective and safer S1P1 receptor agonists are still needed clinically. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a compound represented by formula IR or IS, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester, prodrug, homologue, hydrate or solvate thereof, in view of the deficiencies in the prior art.

[0006] The technical problem that the present invention also solves is to provide a pharmaceutical composition containing the compound represented by the above formula IR or IS, or its stereoisomer, or its pharmaceutically acceptable salt, ester, prodrug, homologue, hydrate or solvate.

[0007] The technical problem to be further solved by the present invention is to provide a compound represented by the above formula IR or IS, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester, prodrug, homologue, hydrate or solvate thereof, or an application of the above pharmaceutical composition.

[0008] In order to solve the above-mentioned first technical problem, the present invention discloses a compound represented by formula IR or IS, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester, prodrug, homologue, hydrate or solvate thereof,

[0009]

[0010] in,

[0011] R1 is a C1-C6 alkyl group substituted by halogen;

[0012] R2 and R3 are independently selected from hydrogen, C1-C6 alkyl, C1-C6 acyl, C1-C6 alkylsulfonyl, wherein the C1-C6 alkyl, C1-C6 acyl, C1-C6 alkylsulfonyl are independently unsubstituted or substituted with hydroxyl, amino, carboxyl, C1-C4 alkoxy, -NR4R5, C1-C4 alkyl; R4 and R5 are independently selected from C1-C6 alkyl.

[0013] In some embodiments, R1 is a C1-C4 alkyl group substituted by fluorine; preferably, R1 is difluoromethyl, difluoroethyl or trifluoroethyl.

[0014] In some embodiments, R2 and R3 are independently selected from hydrogen, C1-C4 alkyl, C1-C4 acyl, C1-C4 alkylsulfonyl, wherein the C1-C4 alkyl, C1-C4 acyl, C1-C4 alkylsulfonyl are independently unsubstituted or substituted with hydroxyl, amino, carboxyl, C1-C4 alkoxy, -NR4R5, C1-C4 alkyl, and R4 and R5 are independently selected from C1-C4 alkyl; preferably, R2 and R3 are independently wherein R2 and R3 are independently selected from hydrogen, C1-C4 alkyl, C1-C4 acyl, C1-C4 alkylsulfonyl, wherein the C1-C4 alkyl, C1-C4 acyl, C1-C4 alkylsulfonyl are independently unsubstituted or substituted by hydroxy, amino, carboxyl, C1-C4 alkoxy, -NR4R5, R4 and R5 are independently selected from methyl or ethyl; more preferably, R2 and R3 are independently selected from hydrogen, methyl, hydroxyethyl, acetyl, methylsulfonyl, ethylsulfonyl,

[0015] In some embodiments, the compound is any one of the following structures:

[0016]

[0017]

[0018] In some embodiments, the pharmaceutically acceptable salt is hydrochloride, hydrobromide, sulfate, phosphate, nitrate, acetate, maleate, fumarate, citrate, tartrate, methanesulfonate, p-toluenesulfonate or benzenesulfonate, preferably hydrochloride.

[0019] The use of the above compounds in the preparation of activators or agonists of sphingosine 1-phosphate receptor subtype 1 is also within the protection scope of the present invention.

[0020] Unless otherwise specified, the following terms used in this application have the following meanings. A particular term should not be considered to be uncertain or unclear in the absence of a special definition, but should be understood according to the common meaning in the art. When a trade name appears in this article, it is intended to refer to the corresponding commodity or its active ingredient.

[0021] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent as long as the valence state of the particular atom is normal and the compound after the substitution is stable.

[0022] In this article, C n ~C m , means that the moiety has an integer number of carbon atoms in a given range. For example, "C1-C6" means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms. For example, C1-C3 means that the group can have 1 carbon atom, 2 carbon atoms, or 3 carbon atoms.

[0023] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 2 R's, each R has an independent option.

[0024] When the number of a linking group is 0, such as -(CH2)0-, it means that the linking group is a covalent bond.

[0025] When one of the variables is selected from a covalent bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a covalent bond, it means that the structure is actually AZ.

[0026] When a substituent's bond crosses two atoms in a ring, the substituent may be bonded to any atom in the ring. It means that it can be substituted at any position on the cyclohexyl group or cyclohexadiene.

[0027] The term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

[0028] The term "hydroxy" refers to an -OH group.

[0029] The term "amino" refers to a -NH2 group.

[0030] The term "cyano" refers to a -CN group.

[0031] The term "nitro" refers to a -NO2 group.

[0032] The term "carboxy" refers to a -COOH group.

[0033] The term "alkyl" refers to a group of the formula C n H 2n+1 The alkyl group may be straight chain or branched. For example, the term "C 1- "C6 alkyl" refers to an alkyl group containing 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc.). Similarly, the alkyl portion (i.e., alkyl) of alkoxy, alkylamino, dialkylamino, alkylsulfonyl, and alkylthio has the same definition as above. For another example, the term "C 1- The term "C3 alkyl" refers to an alkyl group containing 1 to 3 carbon atoms (eg, methyl, ethyl, propyl and isopropyl).

[0034] The term "alkoxy" refers to an alkyl group as described above with a specific number of carbon atoms connected by an oxygen bridge. C1-C6 alkoxy groups include C1, C2, C3, C4, C5 and C6 alkoxy groups. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy and S-pentoxy.

[0035] The term "acyl" refers to a group containing a carbonyl moiety, wherein the group is bonded through a carbonyl carbon atom. The carbonyl carbon atom is also bonded to another carbon atom, which may be part of an alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl group, and the like. In the specific case where the carbonyl is attached to hydrogen, the group is a "formyl", which is an acyl group as that term is defined herein. Unless otherwise specified, an acyl group may include 0 to about 12-20 additional carbon atoms attached to the carbonyl group. Acyl groups as referred to herein may contain double or triple bonds. Acryloyl is an example of an acyl group. Acyl groups as referred to herein may also contain heteroatoms. Nicotinoyl (pyridyl-3-carbonyl) is an example of an acyl group as referred to herein. Other examples include acetyl, benzoyl, phenylacetyl, pyridylacetyl, cinnamoyl, and acryloyl, and the like. When the group containing the carbon atom attached to the carbonyl carbon atom contains a halogen, the group is referred to as a "haloacyl" group. An example is trifluoroacetyl.

[0036] The term "alkylsulfonyl" refers to a -SO2-alkyl group.

[0037] The term "treatment" means administering the compound or formulation described herein to improve or eliminate a disease or one or more symptoms associated with the disease, and includes:

[0038] i. To inhibit a disease or disease state, i.e. to curb its development;

[0039] ii. To alleviate a disease or condition, even if the disease or condition regresses.

[0040] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0041] The term "pharmaceutically acceptable salt" refers to salts of compounds of the present application, prepared from compounds with specific substituents found in the present application and relatively non-toxic acids or bases. When the compounds of the present application contain relatively acidic functional groups, base addition salts can be obtained by contacting a neutral form of such compounds with a suitable base. When the compounds of the present application contain relatively basic functional groups, acid addition salts can be obtained by contacting a neutral form of such compounds with a suitable acid. Certain specific compounds of the present application contain basic and acidic functional groups, and thus can be converted into either base or acid addition salts.

[0042] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present application or their salts and pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present application to an organism.

[0043] The word "comprise" or "comprises" and its English variations such as comprises or comprising should be construed in an open and non-exclusive sense, ie, "including but not limited to".

[0044] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All of these isomers and their mixtures are included within the scope of the present invention.

[0045] Unless otherwise indicated, "(D)" or "(+)" indicates dextrorotatory, "(L)" or "(-)" indicates levorotatory, and "(DL)" or "(±)" indicates racemic.

[0046] Unless otherwise specified, the key is a solid wedge. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed key To indicate the relative configuration of a stereocenter, use a wavy line Indicates a wedge-shaped solid key or dotted wedge key Or use a wavy line Represents a straight solid bond and straight dashed key

[0047] Optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished by using chromatography, which uses a chiral stationary phase and is optionally combined with a chemical derivatization method (for example, a carbamate is generated from an amine).

[0048] The compounds of the present invention may contain non-natural proportions of atomic isotopes on one or more atoms constituting the compound. For example, compounds may be labeled with radioactive isotopes, such as tritium (3H), iodine-125 (125I) or C-14 (14C). For another example, deuterated drugs may be formed by replacing hydrogen with heavy hydrogen, such as d3-methyl, which means that all three hydrogen atoms on the methyl group are replaced by deuterium atoms. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have the advantages of reducing toxic side effects, increasing drug stability, enhancing therapeutic effects, and extending the biological half-life of drugs. All isotopic composition changes of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.

[0049] The present application also includes isotope-labeled compounds of the present application that are identical to those described herein, but one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number commonly found in nature. Examples of isotopes that can be incorporated into compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2H, 3H, 11C, 13C, 14C, 13N, 15N, 15O, 17O, 18O, 31P, 32P, 35S, 18F, 123I, 125I, and 36Cl, etc.

[0050] Certain isotopically labeled compounds of the present invention (e.g., those labeled with 3H and 14C) can be used in compound and / or substrate tissue distribution analysis. Tritiated (i.e., 3H) and carbon-14 (i.e., 14C) isotopes are particularly preferred due to their ease of preparation and detectability. Positron emitting isotopes, such as 15O, 13N, 11C, and 18F can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the present invention can generally be prepared by replacing an isotopically labeled reagent with an isotopically labeled reagent by the following procedures similar to those disclosed in the schemes and / or embodiments below.

[0051] In addition, the use of heavier isotopes such as deuterium (i.e. 2 H)) substitution may provide certain therapeutic advantages arising from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and may therefore be preferred in certain circumstances, wherein deuterium substitution may be partial or complete, partial deuterium substitution means that at least one hydrogen is replaced by at least one deuterium, and all such forms of compounds are included within the scope of the present application.

[0052] The compounds of the present application may be asymmetric, for example, having one or more stereoisomers. Unless otherwise indicated, all stereoisomers are included, such as enantiomers and diastereomers. The compounds of the present application containing asymmetric carbon atoms can be separated in optically pure form or racemic form. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral raw materials or chiral reagents.

[0053] The pharmaceutical composition of the present application can be prepared by combining the compound of the present application with suitable pharmaceutically acceptable excipients, for example, it can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols, etc.

[0054] Typical routes of administration of the compounds of the present application or their pharmaceutically acceptable salts or their pharmaceutical compositions include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0055] The pharmaceutical composition of the present application can be manufactured by methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, sugar-coated pill making methods, grinding methods, emulsification methods, freeze-drying methods, etc.

[0056] The therapeutic dose of the compounds of the present application may be determined, for example, based on the specific use of the treatment, the method of administering the compound, the health and condition of the patient, and the judgment of the prescribing physician. The ratio or concentration of the compounds of the present application in the pharmaceutical composition may not be fixed, depending on a variety of factors, including dosage, chemical properties (e.g., hydrophobicity), and route of administration. For example, the compounds of the present application may be provided by a physiological buffered aqueous solution containing about 0.1 to 10% (g / mL) of the compound for parenteral administration. Some typical dosage ranges are about 1 μg / kg to about 1 g / kg body weight / day. In certain embodiments, the dosage range is about 0.01 mg / kg to about 100 mg / kg body weight / day. The dosage is likely to depend on such variables as the type and degree of development of the disease or condition, the general health status of the specific patient, the relative biological efficacy of the selected compound, the excipient formulation, and its route of administration. The effective dose can be obtained by extrapolation of a dose-response curve derived from an in vitro or animal model test system.

[0057] The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitution methods well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present application.

[0058] The chemical reactions of the specific embodiments of the present application are carried out in a suitable solvent, which must be suitable for the chemical changes of the present application and the reagents and materials required. In order to obtain the compounds of the present application, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction processes based on the existing embodiments.

[0059] Beneficial effects:

[0060] (1) The present invention provides a 1-sphingosine phosphate receptor subtype 1 modulator having a polyfluorosubstituted alkyl group, wherein the compound has high agonist activity for 1-sphingosine phosphate receptor subtype 1, EC 50 In 10 -9 ~10 -10 Within the range of M.

[0061] (2) The regulator prepared by the present invention has good S1PR1 agonist activity. DETAILED DESCRIPTION

[0062] Example 1

[0063] Synthesis of 2-(difluoromethoxy)-5-(3-((1S)-1-((2-hydroxyethyl)amino)-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)benzonitrile hydrochloride (IS-3), the structural formula of which is as follows:

[0064]

[0065] The synthetic route is as follows:

[0066]

[0067]

[0068] The synthesis method is as follows:

[0069] Step 1: Synthesis of compound 1-1

[0070] Methyl 3-bromo-4-hydroxybenzoate (10 g, 0.043 mol), sodium difluorochloroacetate (19.4 g, 0.127 mol), cesium carbonate (28.2 g, 0.087 mol) and N,N-dimethylformamide (DMF, 60 mL) were added to a three-necked round-bottom flask, and stirred at 110°C for 16 h under nitrogen protection. The reaction solution was cooled to room temperature, filtered, and the filter cake was washed with ethyl acetate (EA, 80 mL). 600 mL of water phase was added to the organic phase, and the aqueous phase was extracted with EA (80 mL × 3). The organic phases were combined, washed with water (50 mL × 3), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a yellow oily crude product, which was purified by column chromatography (PE: EA = 100: 1) to obtain product 1-1 (8.72 g, 71.7%).

[0071] 1 H NMR (400MHz, Chloroform-d) δ8.33(d,J=2.1Hz,1H),8.02(dd,J=8.6,2.1Hz,1H),7.30–7.26(m,1H),6.63(t,J=72.6Hz,1H),3.95(s,3H).

[0072] Step 2: Synthesis of compound 1-2

[0073] Compound 1-1 (8 g, 28.47 mmol) was dissolved in DMF (50 mL), and cuprous cyanide (7.6 g, 84.86 mmol), cuprous iodide (0.5 g, 2.632 mmol), and 2-acetylcyclohexanone (0.03 g, 0.214 mmol) were added in sequence, and the mixture was stirred at 120°C for 16 h under nitrogen protection. The reaction solution was cooled to room temperature, diluted with EA (50 mL), filtered, and the filter cake was rinsed with EA (150 mL). Water (250 mL) was added to the filtrate, and the phases were separated. The aqueous phase was extracted with EA (50 mL × 3), and the organic phases were combined, washed with water (20 mL × 3), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain yellow solid compound 1-2 (5.72 g, 88.4%).

[0074] Step 3: Synthesis of Compound 1-3

[0075] Compound 1-2 (5.17 g, 22.76 mmol) was dissolved in 1,4-dioxane (20 mL), and lithium hydroxide monohydrate (2.9 g, 69.11 mmol) was dissolved in 20 mL of water and slowly added to the reaction solution, and stirred at room temperature for 2 h. 1N HCl was added to the reaction solution under ice-water bath conditions to adjust the pH to 1, and filtered to obtain white solid compound 1-3 (4.5 g, 92.8%).

[0076] 1 H NMR (400MHz, DMSO-d6) δ13.53(s,1H),8.37(d,J=2.1Hz,1H),8.29(dd,J=8.8,2.2Hz,1H),7.77–7.31(m,2H).

[0077] Step 4: Synthesis of Compound 1-4

[0078] Dissolve 4-bromoindanone (80 g, 0.379 mol) in DMF (200 mL), add cuprous cyanide (102 g, 1.14 mol), cuprous iodide (7.2 g, 0.038 mol), and 2-acetylcyclohexanone (1.8 g, 0.013 mol) in sequence, and stir the reaction at 120°C under nitrogen protection for 21 h. The reaction solution was cooled to room temperature, diluted with dichloromethane (DCM, 250 mL), filtered, the filter cake was rinsed with DCM (500 mL), the filtrate was added with water (1.6 L) for phase separation, the aqueous phase was extracted with DCM (250 mL × 3), the organic phases were combined, washed with water (200 mL × 3), and concentrated under reduced pressure to obtain a green solid crude product. The crude product was heated to reflux with 150 mL (ethanol: methanol = 4: 1) and stirred for 1 h. After cooling to room temperature, stirring was continued for 30 min and filtered to obtain a yellow solid compound 1-4 (43.1 g, 72.4%).

[0079] 1 H NMR (400MHz, Chloroform-d) δ7.97(d,J=7.7Hz,1H),7.88(d,J=7.2Hz,1H),7.52(t,J=7.6Hz,1H),3.37~3.29(m,2H),2.83~2.76(m,2H).

[0080] Step 5: Synthesis of Compound 1-5

[0081] Compound 1-4 (52.9 g, 0.337 mol) was dissolved in toluene (250 mL), and (s)-(-)-tert-butylsulfenamide (44.9 g, 0.370 mol) and tetraethyl titanate (115 g, 0.504 mol) were added in sequence. The reaction was stirred at 60 ° C for 4 h under nitrogen protection. The reaction solution was diluted with tetrahydrofuran (THF, 400 mL) and cooled to -30 ° C. Sodium borohydride (45.9 g, 1.21 mol) was added in batches, and the temperature did not exceed -25 ° C during the period. After stirring for one hour, the reaction solution was slowly heated to 0 ° C and stirred for 16 h. The reaction solution was quenched by adding saturated potassium sodium tartrate (400 mL) and saturated sodium chloride solution (100 mL) under ice bath. A large amount of light yellow solid was precipitated, EA (550 mL) was added and stirred for 2 h, filtered, the filter cake was rinsed with EA (300 mL), the filtrate was separated, the aqueous phase was extracted with EA (250 mL × 3), the organic phases were combined, the organic phases were washed once with saturated sodium bicarbonate (100 mL), and then washed with saturated sodium chloride water (100 mL × 2), and concentrated to obtain yellow solid compound 1-5 (55.1 g, 62.5%). The crude product was used directly in the next step without further purification.

[0082] Step 6: Synthesis of Compound 1-6

[0083] Compound 1-5 (55.1 g, 0.21 mol) was dissolved in a methanol solution of HCl (w / w 13.5%, 90 mL) and 1,4-dioxane (60 mL), and the mixture was stirred at room temperature for about 3 h. The reaction solution was filtered, the filter cake was rinsed with methanol (180 mL), and the filtrate was concentrated under reduced pressure to obtain a green sticky solid crude product (38 g). Acetonitrile (180 mL) was added to the crude product and heated to reflux and stirred for 2 h, then slowly cooled to room temperature, stirred in an ice bath for 10 min, and filtered to obtain a yellow-brown solid 1-6 (18.1 g, 44.3%).

[0084] 1 H NMR(400MHz, Deuterium Oxide)δ7.79(dd,J=7.8,4.1Hz,2H),7.51(t,J=7.8Hz,1H),4.97(dd,J=8.1,5.0Hz ,1H),3.40~3.28(m,1H),3.26~3.14(m,1H),2.80~2.66(m,1H),2.30~2.17(m,1H).

[0085] Step 7: Synthesis of Compound 1-7

[0086] Compound 1-6 (3.13 g, 0.016 mol) was dissolved in DCM (10 mL), stirred, and triethylamine (3.33 g, 0.033 mol) was added dropwise under ice bath. Di-tert-butyl dicarbonate (4.89 g, 0.022 mol) was dissolved in DCM (10 mL) and slowly added dropwise to the reaction solution. The reaction was stirred at room temperature for 2 h. The reaction solution was diluted with DCM (30 mL), and then washed with saturated sodium chloride solution (30 mL × 3). The organic phase was dried with anhydrous sodium sulfate and filtered. After adding DCM to the filtrate to a total volume of 70 mL, 1.2 g of activated carbon was added and stirred under hot reflux for 30 min. After that, it was filtered while hot (diatomaceous earth), and the filtrate was concentrated to obtain yellow-green solid compound 1-7 (3.74 g, 90.1%). The crude product was used for the next step without further purification.

[0087] Step 8: Synthesis of Compound 1-8

[0088] Compound 1-7 (3.33 g, 12.9 mmol) was dissolved in DMF (25 mL), and under nitrogen protection, NaH (2.52 g, 63 mmol, NaH purity of 60%) was added in batches at 0°C, and the internal temperature was kept at 0°C and stirred for 1 h. After 1 h, (2-bromoethoxy)-tert-butyldimethylsilane (6.38 g, 26.7 mmol) was added and stirred for 5 min, and then the reaction was stirred at room temperature for 2 h. The reaction solution was added with saturated sodium bicarbonate solution to quench the reaction until no bubbles were generated, and water (100 mL) was added to dilute it, and it was extracted with EA (30 mL×3), and the organic phases were combined, washed with saturated sodium chloride solution (20 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a brown oily crude product, which was purified by column chromatography [eluent: petroleum ether: ethyl acetate (40:1)] to obtain a colorless oily compound 1-8 (3.15 g, 58.7%).

[0089] 1 H NMR(400MHz,Chloroform-d)δ7.52(d,J=7.6Hz,1H),7.41(d,J=23.5Hz,1H),7.31(d,J=7.6Hz,1H),5.50(d,J=196.1Hz,1H),3.96 –3.40(m,3H),3.34–3.14(m,2H),3.11–2.82(m,1H),2.51(s,1H),2.28(d,J=37.8Hz,1H),1.26(s,9H),0.88(s,9H),0.08(s,6H).

[0090] Step 9: Synthesis of Compound 1-9

[0091] Compound 1-8 (2.4 g, 5.77 mmol) was dissolved in ethanol (20 mL), and hydroxylamine hydrochloride (1.2 g, 17.3 mmol) and triethylamine (1.75 g, 17.3 mmol) were added in sequence, and the mixture was stirred under reflux for 3 h under nitrogen protection. The reaction solution was concentrated and redissolved in DCM (80 mL), washed with water (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a light green semi-solid compound 1-9 (2.33 g, 89.6%).

[0092] 1 H NMR (400MHz, DMSO-d6) δ9.53(s,1H),7.37(d,J=7.6Hz,1H),7.22(t,J=7.6Hz,1H),7.08(d,J=21.2Hz,1H),5.68(d,J=6.9Hz,2H),5.55 (s,1H),5.24(s,1H),3.61(d,J=33.6Hz,3H),3.18~2.75(m,5H),2.13(d,J=117.6Hz,3H),1.45(s,5H),1.31~1.17(m,5H),0.83(s,9H).

[0093] Step 10: Synthesis of Compound 1-10

[0094] Compound 1-3 (0.16 g, 0.751 mmol) was dissolved in DMF (5 mL), and 1-hydroxybenzotriazole (0.24 g, 1.78 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.46 g, 2.396 mmol), and triethylamine (0.25 g, 2.48 mmol) were added in sequence, and stirred at room temperature for 1 h. Compound 1-9 (0.3 g, 0.666 mmol) was dissolved in DMF (5 mL) and added to the reaction system, and stirred at room temperature for 3 h. The reaction solution was diluted with saturated saline (80 mL), extracted with EA (20 mL × 3), and the organic phases were combined and washed once with saturated sodium bicarbonate solution (8 mL), and then washed with saturated sodium chloride solution (10 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 0.46 g of yellow oil. The crude product was redissolved in DMF (5 mL) and stirred at 85 ° C overnight. The reaction solution was cooled to room temperature and diluted with water (50 mL), extracted with EA (8 mL × 3), and the organic phases were combined and washed once with saturated sodium bicarbonate solution (5 mL), and then washed with saturated sodium chloride solution (5 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a yellow oil compound 1-10 (0.4 g, 95.6%). The crude product was used directly in the next step without further purification.

[0095] Step 11: Synthesis of compound IS-3

[0096] A methanol solution of HCl (w / w 13.5%, 6 mL) was added to compound 1-10 (0.13 g, 0.207 mmol), and the mixture was stirred at 50°C for 2 h. The reaction solution was concentrated under reduced pressure, and then stirred with methanol (12 mL) for 1 h and filtered to obtain a white solid compound IS-3 (0.07 g, 75.3%).

[0097] 1 H NMR (400MHz, DMSO-d6) δ9.43(d,J=24.4Hz,2H),8.75(d,J=2.2Hz,1H),8.57(dd,J=8.9,2.2Hz,1H ),8.17(dd,J=7.8,1.0Hz,1H),8.04(d,J=7.6Hz,1H),7.75(d,J=8.8Hz,1H),7.66(t,J=71.8Hz,1H ),7.58(t,J=7.7Hz,1H),5.29(t,J=5.2Hz,1H),4.93(s,1H),3.74(q,J=5.2Hz,2H),3.59–3.43(m, 1H),3.32–3.21(m,1H),3.04(d,J=23.9Hz,2H),2.62–2.53(m,1H),2.39–2.26(m,1H).MS(ESI)m / z calcd for C 21 H 19 F2N4O3[M+H]+413.14,found 413.57.

[0098] Example 2

[0099] Synthesis of 5-(3-((1S)-1-amino-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)-2-(difluoromethoxy)benzonitrile hydrochloride (IS-1), the structural formula of which is as follows:

[0100]

[0101] The synthetic route is as follows:

[0102]

[0103] The synthesis method is as follows:

[0104] Step 1: Synthesis of compound 2-1

[0105] Compound 1-7 (1 g, 3.87 mmol) was dissolved in ethanol (10 mL), and hydroxylamine hydrochloride (0.81 g, 11.66 mmol) and triethylamine (1.2 g, 11.86 mmol) were added in sequence, and the mixture was stirred under reflux for 3 h under nitrogen protection. The reaction solution was concentrated and redissolved in DCM (30 mL), washed with water (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 1.37 g of a yellow oily crude product, which was purified by column chromatography (PE: EA = 2: 1) to obtain product 2-1 (1.1 g, 97.3%).

[0106] 1 H NMR(400MHz,Chloroform-d)δ7.78(s,1H),7.38-7.25(m,1H),7.27–7.00(m,3H),5.19(d,J=8.5Hz,1H),5.00(d ,J=8.9Hz,1H),4.83(s,1H),3.26–2.99(m,1H),2.91-2.87(m,1H),2.54(s,1H),1.85–1.64(m,1H),1.52(s,9H).

[0107] Step 2: Synthesis of compound 2-2

[0108] Compound 1-3 (0.34 g, 1.6 mmol) was dissolved in DMF (5 mL), and 1-hydroxybenzotriazole (0.45 g, 3.3 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1 g, 5.22 mmol), and triethylamine (0.54 g, 5.35 mmol) were added in sequence, and stirred at room temperature for 1 h. Compound 2-1 (0.42 g, 1.4 mmol) was dissolved in DMF (5 mL) and added to the reaction system, and stirred at room temperature for 3 h. The reaction solution was diluted with saturated saline (80 mL), extracted with EA (20 mL × 3), and the organic phases were combined and washed once with saturated sodium bicarbonate solution (8 mL), and then washed with saturated sodium chloride solution (10 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 0.74 g of yellow solid. The crude product was redissolved in DMF (5 mL), and stirred at 85 ° C for overnight. The reaction solution was cooled to room temperature and diluted with water (50 mL), extracted with EA (8 mL × 3), and the organic phases were combined and washed once with a saturated sodium bicarbonate solution (5 mL), and then washed with a saturated sodium chloride solution (5 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a yellow solid compound 2-2 (0.64 g, 94.7%). The crude product was used directly in the next step without further purification.

[0109] Step 3: Synthesis of compound IS-1

[0110] A methanol solution of HCl (w / w 13.5%, 6 mL) was added to compound 2-2 (0.21 g, 0.448 mmol), and the mixture was stirred at 50°C for 2 h. The reaction solution was concentrated under reduced pressure, and then stirred with methanol (12 mL) for 1 h and filtered to obtain a white solid compound IS-1 (0.13 g, 72.2%).

[0111] 1 H NMR (400MHz, DMSO-d6) δ8.74(d,J=2.2Hz,1H),8.63(s,2H),8.60–8.53(m,1H),8.14(dd,J=7.8,1.1Hz,1H),7.90(d,J=7.6Hz,1 H),7.84–7.46(m,3H),4.82(s,1H),3.55–3.42(m,1H),3.28–3.23(m,1H),2.62–2.52(m,1H),2.18–2.05(m,1H).HRMS(ESI)m / z calcdfor C 19 H 15 F2N4O2[M+H] + 369.1163, found 369.1171.

[0112] Example 3

[0113] Synthesis of N-((1S)-4-(5-(3-cyano-4-(difluoromethoxy)phenyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)acetamide (IS-2) having the following structural formula:

[0114]

[0115] Its synthetic route is as follows:

[0116]

[0117] The synthesis method is as follows:

[0118] Step 1: Synthesis of compound IS-2

[0119] Compound IS-1 (0.044 g, 0.109 mmol) was dissolved in DCM (5 mL), and acetic anhydride (0.017 g, 0.168 mmol) and triethylamine (0.017 g, 0.168 mmol) were added in sequence, and the mixture was reacted at room temperature for 3 h. The reaction solution was diluted with DCM (40 mL), washed once with saturated sodium bicarbonate solution (8 mL), and then washed with saturated sodium chloride solution (5 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain white solid compound IS-2 (0.032 g, 71.7%).

[0120] 1 H NMR (400MHz, DMSO-d6) δ8.74(d,J=2.2Hz,1H),8.56(dd,J=8.8,2.2Hz,1H),8.31(d,J=8.2Hz,1H),8.01(p,J=3.8Hz,1H),7.82–7.62(m,2H),7.44 (d,J=4.3Hz,2H),5.36(q,J=8.0Hz,1H),3.43–3.36(m,1H),3.18–3.05(m,1H),2.48–2.42(m,1H),1.89(s,3H),1.87–1.78(m,1H).HRMS(ESI)m / z calcd for C 21 H 17 F2N4O3[M+H] + 411.1269, found 411.1270.

[0121] Example 4

[0122] Synthesis of 5-(3-((1S)-1-(bis(2-hydroxyethyl)amino)-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)-2-(difluoromethoxy)benzonitrile (IS-4), the structural formula of which is as follows:

[0123]

[0124] The synthetic route is as follows:

[0125]

[0126] Step 1: Synthesis of compound IS-4

[0127] Compound IS-1 (0.3 g, 0.741 mmol) was dissolved in H2O (5 mL), and (30 wt.% in THF) ethylene oxide (20 mL), triethylamine (0.11 g, 1.089 mmol) were added in sequence, and the mixture was reacted at room temperature for 26 h. The reaction solution was diluted with EA (20 mL) and washed with saturated sodium chloride solution (5 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM: MEOH = 35: 1) to obtain a white solid compound IS-4 (110 mg, 32.4%).

[0128] 1H NMR(400MHz,Chloroform-d)δ8.58(d,J=2.1Hz,1H),8.47(dd,J=8.8,2.2Hz,1H),8.12(d ,J=7.7Hz,1H),7.58(t,J=8.5Hz,2H),7.44(t,J=7.6Hz,1H),6.79(t,J=70.8Hz,1H),5.32 (s,1H),4.68(t,J=7.5Hz,1H),3.82–3.70(m,2H),3.68–3.57(m,2H),3.50–3.40(m,1H),3 .29–3.14(m,1H),2.84–2.69(m,4H),2.43–2.33(m,1H),2.18–2.07(m,2H).HRMS(ESI)m / z calcd for C 23 H 23 F2N4O4[M+H] + 457.1687, found 457.1686.

[0129] Example 5

[0130] Synthesis of 5-(3-((1S)-1-((2-hydroxyethyl)amino)-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)-2-(2,2,2-trifluoroethoxy)benzonitrile hydrochloride (IS-11), the structural formula of which is as follows:

[0131]

[0132] Its synthetic route is as follows:

[0133]

[0134] The synthesis method is as follows:

[0135] Step 1: Synthesis of compound 5-1

[0136] Methyl 3-bromo-4-hydroxybenzoate (10 g, 0.043 mol), trifluoroethyl iodide (23 g, 0.1095 mol), potassium carbonate (15 g, 0.1085 mol) and N,N-dimethylformamide (DMF, 50 mL) were added to a three-necked round-bottom flask, and the mixture was stirred at 100°C for 15 h under nitrogen protection. The reaction solution was cooled to room temperature, filtered, and the filter cake was washed with ethyl acetate (EA, 50 mL). 400 mL of water phase was added to the organic phase, and the aqueous phase was extracted with EA (50 mL × 3). The organic phases were combined, washed with water (30 mL × 3), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a white solid crude product, which was purified by column chromatography (PE: EA = 30: 1) to obtain product 5-1 (4.85 g, 36.3%).

[0137] 1 H NMR (400MHz, Chloroform-d) δ8.30(d,J=2.1Hz,1H),8.02(dd,J=8.6,2.1Hz,1H),6.95(d,J=8.6Hz,1H),4.49(q,J=7.9Hz,2H),3.93(s,3H).

[0138] Step 2: Synthesis of compound 5-2

[0139] Compound 5-1 (4.6 g, 14.7 mmol) was dissolved in DMF (35 mL), and cuprous cyanide (3.9 g, 43.5 mmol), cuprous iodide (0.28 g, 1.47 mmol), and 2-acetylcyclohexanone (0.06 g, 0.428 mmol) were added in sequence, and the mixture was stirred at 120°C for 16 h under nitrogen protection. The reaction solution was cooled to room temperature, diluted with EA (25 mL), filtered, and the filter cake was rinsed with EA (150 mL). Water (250 mL) was added to the filtrate, and the phases were separated. The aqueous phase was extracted with EA (50 mL × 3), and the organic phases were combined, washed with water (20 mL × 3), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a yellow solid compound 5-2 (2.7 g, 71%). 1 H NMR (400MHz, Chloroform-d) δ8.34(d,J=2.1Hz,1H),8.28(dd,J=8.8,2.2Hz,1H),7.06(d,J=8.8Hz,1H),4.59(q,J=7.7Hz,2H),3.96(s,3H).

[0140] Step 3: Synthesis of compound 5-3

[0141] Compound 5-2 (2.37 g, 9.14 mmol) was dissolved in 1,4-dioxane (10 mL), and lithium hydroxide monohydrate (1.5 g, 35.8 mmol) was dissolved in 10 mL of water and slowly added to the reaction solution, and stirred at room temperature for 2 h. 1N HCl was added to the reaction solution under ice-water bath conditions to adjust the pH to 1, and filtered to obtain white solid compound 5-3 (2.1 g, 93.8%).

[0142] 1 H NMR (400MHz, Chloroform-d) δ12.55 (s, 1H), 8.42 (d, J = 2.2Hz, 1H), 8.35 (dd, J = 8.9, 2.2Hz, 1H), 7.11 (d, J = 8.9Hz, 1H), 4.61 (q, J = 7.7Hz, 2H).

[0143] Step 4: Synthesis of compound 5-4

[0144] Compound 5-3 (0.35 g, 1.43 mmol) was dissolved in DMF (5 mL), and 1-hydroxybenzotriazole (0.41 g, 3.03 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.77 g, 4.02 mmol), and triethylamine (0.42 g, 4.16 mmol) were added in sequence, and stirred at room temperature for 1 h. Compound 1-9 (0.5 g, 1.11 mmol) was dissolved in DMF (5 mL) and added to the reaction system, and stirred at room temperature for 3 h. The reaction solution was diluted with saturated saline (100 mL), extracted with EA (15 mL × 3), and the organic phases were combined and washed once with saturated sodium bicarbonate solution (15 mL), and then washed with saturated sodium chloride solution (15 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 0.83 g of light yellow solid. The crude product was redissolved in DMF (8 mL) and stirred at 85 ° C overnight. The reaction solution was cooled to room temperature and diluted with water (80 mL), extracted with EA (8 mL × 3), and the organic phases were combined and washed once with saturated sodium bicarbonate solution (10 mL), and then washed with saturated sodium chloride solution (10 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a yellow oil compound 5-4 (0.71 g, 97.3%). The crude product was used directly in the next step without further purification.

[0145] Step 5: Synthesis of compound IS-11

[0146] A methanol solution of HCl (w / w 13.5%, 10 mL) was added to compound 5-4 (0.71 g, 1.08 mmol), and the mixture was stirred at 50°C for 2 h. The reaction solution was concentrated under reduced pressure, and then stirred with methanol (12 mL) for 1 h and filtered to obtain a white solid compound IS-11 (0.28 g, 53.8%).

[0147] 1 H NMR (400MHz, DMSO-d6) δ9.18(s,2H),8.64(d,J=2.2Hz,1H),8.53(dd,J=9.0,2.2Hz,1H ),8.18(d,J=7.7Hz,1H),7.96(d,J=7.6Hz,1H),7.68(d,J=9.0Hz,1H),7.59(t,J=7.7Hz ,1H),5.19(q,J=8.6Hz,2H),4.93(s,1H),3.72(t,J=5.3Hz,2H),3.57–3.46(m,2H),3.1 7(s,1H),3.10–2.98(m,2H),2.58–2.54(m,1H),2.34–2.27(m,1H).HRMS(ESI)m / zcalcd for C 22 H 20 F3N4O3[M+H] + 445.1488, found 445.1491.

[0148] Example 6

[0149] Synthesis of 5-(3-((1S)-1-amino-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)-2-(2,2,2-trifluoroethoxy)benzonitrile hydrochloride (IS-9), the structural formula of which is as follows:

[0150]

[0151] Its synthetic route is as follows:

[0152]

[0153] The synthesis method is as follows:

[0154] Step 1: Synthesis of compound 6-1

[0155] Compound 5-3 (1.53 g, 6.24 mmol) was dissolved in DMF (15 mL), and 1-hydroxybenzotriazole (2 g, 14.81 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3 g, 15.65 mmol), and triethylamine (2.12 g, 20.99 mmol) were added in sequence, and stirred at room temperature for 1 h. Compound 2-1 (1.65 g, 5.66 mmol) was dissolved in DMF (15 mL) and added to the reaction system, and stirred at room temperature for 4 h. The reaction solution was diluted with saturated saline (300 mL), extracted with EA (35 mL × 3), and the organic phases were combined and washed once with saturated sodium bicarbonate solution (15 mL), and then washed with saturated sodium chloride solution (15 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 3.3 g of yellow solid. The crude product was redissolved in DMF (20 mL) and stirred at 85 ° C overnight. The reaction solution was cooled to room temperature and diluted with water (200 mL), extracted with EA (25 mL × 3), and the organic phases were combined and washed once with saturated sodium bicarbonate solution (25 mL), and then washed with saturated sodium chloride solution (25 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain yellow solid compound 6-1 (2.8 g, 98.9%). The crude product was used directly in the next step without further purification.

[0156] Step 2: Synthesis of compound IS-9

[0157] A methanol solution of HCl (w / w 13.5%, 20 mL) was added to compound 6-1 (2.8 g, 5.6 mmol) and stirred at 50°C for 2 h. The reaction solution was concentrated under reduced pressure, stirred with methanol (20 mL) for 1 h, and filtered to obtain a white solid compound IS-9 (1.58 g, 64.7%).

[0158] 1 H NMR (400MHz, DMSO-d6) δ8.63(s,1H),8.59(s,2H),8.52(d,J=9.0Hz,1H),8.14(d,J=7.7Hz,1H),7.88(d,J=7.6Hz,1H),7.67(d,J=9.1Hz,1H),7.57(t, J=7.7Hz,1H),5.18(q,J=8.6Hz,2H),4.82(t,J=7.0Hz,1H),3.56–3.43(m,1 H),3.29–3.19(m,1H),2.60–2.54(m,1H),2.18–2.04(m,1H).HRMS(ESI)m / z calcd for C 20 H 16 F3N4O2[M+H] +401.1225, found 401.1216.

[0159] Example 7

[0160] Synthesis of N-((1S)-4-(5-(3-cyano-4-(2,2,2-trifluoroethoxy)phenyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)acetamide (IS-10) having the following structural formula:

[0161]

[0162] Its synthetic route is as follows:

[0163]

[0164] The synthesis method is as follows:

[0165] Step 1: Synthesis of compound IS-10

[0166] Compound IS-9 (0.3 g, 0.687 mmol) was dissolved in DCM (5 mL), and acetic anhydride (0.11 g, 1.077 mmol) and triethylamine (0.1 g, 0.988 mmol) were added in sequence. The reaction mixture was diluted with DCM (60 mL), washed once with saturated sodium bicarbonate solution (10 mL), and then washed with saturated sodium chloride solution (10 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a white solid crude product. The crude product was purified by column chromatography (DCM: MeOH = 50: 1) to obtain a white solid compound IS-10 (0.21 g, 63.6%).

[0167] 1 H NMR(400MHz,Chloroform-d)δ8.49(d,J=2.2Hz,1H),8.42(dd,J=8.9,2.2Hz,1H),8.09 (d,J=7.6Hz,1H),7.49(d,J=7.5Hz,1H),7.40(t,J=7.6Hz,1H),7.18(d,J=8.9Hz,1H), 5.72(d,J=8.6Hz,1H),5.58(q,J=7.9Hz,1H),4.61(q,J=7.7Hz,2H),3.52–3.44(m,1H) ,3.26–3.17(m,1H),2.74–2.65(m,1H),2.07(s,3H),1.94–1.84(m,1H).HRMS(ESI)m / z calcd for C 22 H 18 F3N4O3[M+H] +443.1331, found 443.1312.

[0168] Example 8

[0169] Synthesis of 5-(3-((1S)-1-(bis(2-hydroxyethyl)amino)-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)-2-(2,2,2-trifluoroethoxy)benzonitrile (IS-12), the structural formula of which is as follows:

[0170]

[0171] Its synthetic route is as follows:

[0172]

[0173] The synthesis method is as follows:

[0174] Step 1: Synthesis of compound IS-12

[0175] Compound IS-9 (0.05 g, 0.114 mmol) was dissolved in H2O (5 mL), and (30 wt.% in THF) ethylene oxide (5 mL), triethylamine (0.018 g, 0.174 mmol) were added in sequence, and the mixture was reacted at room temperature for 26 h. The reaction solution was diluted with EA (20 mL) and washed with saturated sodium chloride solution (5 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM: MEOH = 5: 1) to obtain a white solid compound IS-12 (20 mg, 32.8%).

[0176] 1 H NMR(400MHz,Chloroform-d)δ8.50(t,J=1.9Hz,1H),8.42(dd,J=8.8,2.2Hz,1H),8.08(d, J=7.6Hz,1H),7.56(d,J=7.4Hz,1H),7.41(t,J=7.6Hz,1H),7.18(d,J=8.8Hz,1H),4.68–4. 57(m,3H),3.77–3.68(m,2H),3.63–3.57(m,2H),3.47–3.38(m,1H),3.24–3.13(m,1H),2. 79–2.68(m,4H),2.39–2.29(m,2H),2.14–2.04(m,1H),1.28(d,J=19.6Hz,1H).MS(ESI)m / z calcd for C 24 H 24 F3N4O4[M+H] +489.18, found 489.11.

[0177] Example 9

[0178] Synthesis of 2-(2,2-difluoroethoxy)-5-(3-((1S)-1-((2-hydroxyethyl)amino)-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)benzonitrile hydrochloride (IS-7), the structural formula of which is as follows:

[0179]

[0180] Its synthetic route is as follows:

[0181]

[0182]

[0183] The synthesis method is as follows:

[0184] Step 1: Synthesis of compound 9-1

[0185] In a three-necked round-bottom flask, methyl 3-bromo-4-hydroxybenzoate (10 g, 0.043 mol), difluoroethyl bromide (8.67 g, 0.06 mol), potassium carbonate (15 g, 0.1085 mol) and N,N-dimethylformamide (DMF, 50 mL) were added to the mixture, and the mixture was stirred at 100 °C for 15 h under nitrogen protection. The reaction solution was cooled to room temperature, filtered, and the filter cake was washed with ethyl acetate (EA, 50 mL). 400 mL of water phase was added to the organic phase, and the aqueous phase was extracted with EA (50 mL × 3). The organic phases were combined, washed with water (30 mL × 3), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a white solid product 9-1 (12.6 g, 98.7%).

[0186] 1 H NMR (400MHz, Chloroform-d) δ8.28(d,J=2.0Hz,1H),8.01(dd,J=8.6,2.1Hz,1H),6.93(d,J=8.6Hz,1H),6.43–5.97(m,1H),4.42–4.21(m,2H),3.93(s,3H).

[0187] Step 2: Synthesis of compound 9-2

[0188] Compound 9-1 (12.2 g, 41.34 mmol) was dissolved in DMF (30 mL), and cuprous cyanide (11.1 g, 123.9 mmol), cuprous iodide (0.79 g, 4.148 mmol), and 2-acetylcyclohexanone (0.17 g, 1.21 mmol) were added in sequence, and the mixture was stirred at 130°C for 23 h under nitrogen protection. The reaction solution was cooled to room temperature, diluted with EA (30 mL), filtered, and the filter cake was rinsed with EA (150 mL). The filtrate was added with brine (250 mL), phases were separated, and the aqueous phase was extracted with EA (30 mL × 3). The organic phases were combined, washed with water (20 mL × 3), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a yellow-green solid crude product, which was purified by column chromatography (PE: EA = 5: 1) to obtain a white product 9-2 (8.07 g, 80.9%).

[0189] 1 H NMR (400MHz, Chloroform-d) δ8.30(d,J=2.1Hz,1H),8.26(dd,J=8.8,2.2Hz,1H),7.05(d,J=8.8Hz,1H),6.43–6.01(m,1H),4.44–4.35(m,2H),3.95(s,3H).

[0190] Step 3: Synthesis of compound 9-3

[0191] Compound 9-2 (7.77 g, 32.21 mmol) was dissolved in 1,4-dioxane (20 mL), and lithium hydroxide monohydrate (3.54 g, 84.37 mmol) was dissolved in 40 mL of water and slowly added to the reaction solution, and stirred at room temperature for 2 h. 1N HCl was added to the reaction solution under ice-water bath conditions to adjust the pH to 1, and filtered to obtain white solid compound 9-3 (6.5 g, 88.8%).

[0192] 1 H NMR (400MHz, Chloroform-d) δ8.39(d,J=2.2Hz,1H),8.33(dd,J=8.8,2.2Hz,1H),7.09(d,J=8.9Hz,1H),6.40–6.03(m,1H),4.46–4.38(m,2H).

[0193] Step 4: Synthesis of compound 9-4

[0194] Compound 9-3 (0.33 g, 1.453 mmol) was dissolved in DMF (5 mL), and 1-hydroxybenzotriazole (0.4 g, 2.96 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.77 g, 4.02 mmol), and triethylamine (0.42 g, 4.16 mmol) were added in sequence, and stirred at room temperature for 1 h. Compound 1-9 (0.5 g, 1.11 mmol) was dissolved in DMF (5 mL) and added to the reaction system, and stirred at room temperature for 3 h. The reaction solution was diluted with saturated saline (100 mL), extracted with EA (15 mL × 3), and the organic phases were combined and washed once with saturated sodium bicarbonate solution (15 mL), and then washed with saturated sodium chloride solution (15 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 0.77 g of light yellow solid. The crude product was redissolved in DMF (8 mL) and stirred at 85 ° C overnight. The reaction solution was cooled to room temperature and diluted with water (80 mL), extracted with EA (8 mL × 3), and the organic phases were combined and washed once with saturated sodium bicarbonate solution (10 mL), and then washed with saturated sodium chloride solution (10 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a yellow oil compound 9-4 (0.69 g, 97.2%). The crude product was used directly in the next step without further purification.

[0195] Step 5: Synthesis of compound IS-7

[0196] A methanol solution of HCl (w / w 13.5%, 10 mL) was added to compound 9-4 (0.69 g, 1.077 mmol), and the mixture was stirred at 50°C for 2 h. The reaction solution was concentrated under reduced pressure, and then stirred with methanol (12 mL) for 1 h and filtered to obtain a white solid compound IS-7 (0.3 g, 60.2%).

[0197] 1H NMR (400MHz, DMSO-d6) δ9.39(m,2H),8.59(d,J=2.2Hz,1H),8.47(dd,J=9.0,2.3Hz,1H),8.15 (d,J=7.7Hz,1H),8.01(d,J=7.6Hz,1H),7.67–7.53(m,2H),6.67–6.34(m,1H),5.28(t,J=5.2 Hz,1H),4.90(d,J=7.8Hz,1H),4.75–4.64(m,2H),3.73(q,J=5.2Hz,2H),3.55–3.45(m,1H),3 .30–3.21(m,1H),3.02(d,J=25.1Hz,2H),2.59–2.51(m,1H),2.36–2.26(m,1H).HRMS(ESI)m / z calcd for C 22 H 21 F2N4O3[M+H] + 427.1582, found 427.1583.

[0198] Example 10

[0199] Synthesis of 5-(3-((1S)-1-amino-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)-2-(2,2-difluoroethoxy)benzonitrile hydrochloride (IS-5), the structural formula of which is as follows:

[0200]

[0201] Its synthetic route is as follows:

[0202]

[0203] The synthesis method is as follows:

[0204] Step 1: Synthesis of compound 10-1

[0205] Compound 9-3 (2 g, 8.81 mmol) was dissolved in DMF (15 mL), and 1-hydroxybenzotriazole (2.51 g, 18.59 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.7 g, 24.52 mmol), and triethylamine (2.57 g, 25.45 mmol) were added in sequence, and stirred at room temperature for 1 h. Compound 2-1 (2 g, 6.87 mmol) was dissolved in DMF (20 mL) and added to the reaction system, and stirred at room temperature for 3 h. The reaction solution was diluted with saturated saline (300 mL), extracted with EA (35 mL × 3), and the organic phases were combined and washed once with saturated sodium bicarbonate solution (15 mL), and then washed with saturated sodium chloride solution (15 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 3.24 g of yellow-white solid. The crude product was redissolved in DMF (20 mL) and stirred at 85 ° C overnight. The reaction solution was cooled to room temperature and diluted with water (200 mL), extracted with EA (25 mL × 3), and the organic phases were combined and washed once with saturated sodium bicarbonate solution (25 mL), and then washed with saturated sodium chloride solution (25 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain yellow solid compound 10-1 (3.21 g, 97%). The crude product was used directly in the next step without further purification.

[0206] Step 2: Synthesis of compound IS-5

[0207] A methanol solution of HCl (w / w 13.5%, 40 mL) was added to compound 10-1 (3.21 g, 6.66 mmol) and stirred at 50°C for 2 h. The reaction solution was concentrated under reduced pressure, stirred with methanol (20 mL) for 1 h, and filtered to obtain a white solid compound IS-5 (1.8 g, 64.5%).

[0208] 1 H NMR (400MHz, DMSO-d6) δ8.65 (s, 3H), 8.59 (d, J = 2.3Hz, 1H), 8.47 (dd, J = 9.0, 2.3Hz ,1H),8.13(d,J=7.7Hz,1H),7.90(d,J=7.6Hz,1H),7.63(d,J=9.0Hz,1H),7.56(t, J=7.7Hz,1H),6.70–6.29(m,1H),4.81(d,J=7.6Hz,1H),4.76–4.64(m,2H),3.57–3 .41(m,1H),3.30–3.20(m,1H),2.61–2.53(m,1H),2.16–2.06(m,1H).HRMS(ESI)m / z calcd forC 20 H 17F2N4O2[M+H] + 383.1320, found 383.1314.

[0209] Embodiment 11

[0210] Synthesis of N-((1S)-4-(5-(3-cyano-4-(2,2-difluoroethoxy)phenyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)acetamide (IS-6) having the following structural formula:

[0211]

[0212] Its synthetic route is as follows:

[0213]

[0214] Step 1: Synthesis of compound IS-6

[0215] Compound IS-5 (0.3 g, 0.718 mmol) was dissolved in DCM (5 mL), and acetic anhydride (0.11 g, 1.08 mmol) and triethylamine (0.1 g, 0.988 mmol) were added in sequence, and the mixture was reacted at room temperature for 5 h. The reaction solution was dried by rotary evaporation, and then methanol (8 mL) was added and stirred for 2 h, and a white solid compound IS-6 (0.22 g, 66.6%) was obtained by suction filtration.

[0216] 1 H NMR (400MHz, DMSO-d6) δ8.59(t,J=2.4Hz,1H),8.48(dd,J=9.0,2.3Hz,1H),8.31(d ,J=8.2Hz,1H),8.02(p,J=3.8Hz,1H),7.62(d,J=9.0Hz,1H),7.45(d,J=4.5Hz,2H) ,6.72–6.26(m,1H),5.36(q,J=7.9Hz,1H),4.74–4.65(m,2H),3.45–3.36(m,1H),3 .17–3.06(m,1H),2.49–2.43(m,1H),1.90(s,3H),1.89–1.80(m,1H).HRMS(ESI)m / z calcd forC 22 H 19 F2N4O3[M+H] + 425.1425, found 425.1423.

[0217] Example 12

[0218] Synthesis of 5-(3-((1R)-1-(bis(2-hydroxyethyl)amino)-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)-2-(2,2-difluoroethoxy)benzonitrile (IS-8), the structural formula of which is as follows:

[0219]

[0220] Its synthetic route is as follows:

[0221]

[0222] The synthesis method is as follows:

[0223] Step 1: Synthesis of compound IS-8

[0224] Compound IS-5 (0.05 g, 0.12 mmol) was dissolved in H2O (5 mL), and (30 wt.% in THF) ethylene oxide (5 mL), triethylamine (0.017 g, 0.174 mmol) were added in sequence, and the mixture was reacted at room temperature for 28 h. The reaction solution was diluted with EA (20 mL) and washed with saturated sodium chloride solution (5 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM: MEOH = 5: 1) to obtain a white solid compound IS-8 (15 mg, 26.8%).

[0225] 1 H NMR(400MHz,Chloroform-d)δ8.48(t,J=1.8Hz,1H),8.41(dd,J=8.9,2.2Hz,1H),8.09(d,J=7.7Hz,1 H),7.56(d,J=7.4Hz,1H),7.41(t,J=7.6Hz,1H),7.17(d,J=8.9Hz,1H),6.38–6.07(m,1H),4.65(t,J= 7.5Hz,1H),4.47–4.39(m,2H),3.77–3.69(m,2H),3.63–3.58(m,2H),3.47–3.38(m,1H),3.25–3.13(m ,1H),2.77–2.71(m,4H),2.40–2.27(m,2H),2.12–2.06(m,1H),1.28(d,J=20.7Hz,1H).HRMS(ESI)m / z calcd for C 24 H 25 F2N4O4[M+H] + 471.1838, found 471.1847.

[0226] Embodiment 13

[0227] Synthesis of 5-(3-((1R)-1-methanesulfonylamino-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)-2-(2,2-difluoroethoxy)benzonitrile (IS-21), the structural formula of which is as follows:

[0228]

[0229] Its synthetic route is as follows:

[0230]

[0231] The synthesis method is as follows:

[0232] Compound IS-5 (0.3 g, 0.718 mmol) was added to DCM (5 mL), followed by methanesulfonyl chloride (0.12 g, 1.04 mmol) and triethylamine (0.1 g, 1.04 mmol), and the mixture was reacted at room temperature for 5 h. The reaction solution was concentrated, methanol (8 mL) was added, and the mixture was stirred for 2 h, and then filtered to obtain compound IS-21 (0.20 g, 60.7%) as a white solid.

[0233] 1 H NMR (400MHz, DMSO-d6) δ8.59(t,J=2.4Hz,1H),8.48(dd,J=9.0,2.3Hz,1H),8.31(d ,J=8.2Hz,1H),8.02(p,J=3.8Hz,1H),7.62(d,J=9.0Hz,1H),7.45(d,J=4.5Hz,2H) ,6.72–6.26(m,1H),5.36(q,J=7.9Hz,1H),4.74–4.65(m,2H),3.45–3.36(m,1H),3 .17–3.06(m,1H),2.93(s,3H),2.49–2.44(m,1H),1.90–1.82(m,1H).HRMS(ESI)m / z calcd forC 21 H 19 F2N4O4S[M+H] + 461.1095, found 461.1073.

[0234] Embodiment 14

[0235] Synthesis of 5-(3-((1R)-1-ethanesulfonylamino-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)-2-(2,2-difluoroethoxy)benzonitrile (IS-22), the structural formula of which is as follows:

[0236]

[0237] Its synthetic route is as follows:

[0238]

[0239] The synthesis method is as follows:

[0240] Referring to the synthesis method of Example 13, compound IS-22 was prepared using IS-5 and ethanesulfonyl chloride as raw materials.

[0241] 1 H NMR (400MHz, DMSO-d6) δ8.59(t,J=2.4Hz,1H),8.48(dd,J=9.0,2.3Hz,1H),8.31(d,J=8.2 Hz,1H),8.02(p,J=3.8Hz,1H),7.62(d,J=9.0Hz,1H),7.45(d,J=4.5Hz,2H),6.72–6.26(m, 1H),5.36(q,J=7.9Hz,1H),4.74–4.65(m,2H),3.45–3.36(m,1H),3.17–3.06(m,1H),3.05( q,J=8.5Hz,2H)2.49–2.43(m,1H),1.89–1.80(m,1H),1.29(t,J=8.7Hz,3H).HRMS(ESI)m / z calcd for C 22 H 21 F2N4O4S[M+H] + 475.1252, found 475.1260.

[0242] Embodiment 15

[0243] Synthesis of N-((1S)-4-(5-(3-cyano-4-(2,2,2-trifluoroethoxy)phenyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)methanesulfonamide (IS-14), the structural formula of which is as follows:

[0244]

[0245] Its synthetic route is as follows:

[0246]

[0247] The synthesis method is as follows:

[0248] Referring to the synthesis method of Example 13, compound IS-14 was prepared using IS-9 and methanesulfonyl chloride as raw materials.

[0249] 1H NMR(400MHz,Chloroform-d)δ8.49(d,J=2.2Hz,1H),8.42(dd,J=8.9,2.2Hz,1H),8.09 (d,J=7.6Hz,1H),7.49(d,J=7.5Hz,1H),7.40(t,J=7.6Hz,1H),7.18(d,J=8.9Hz,1H), 5.72(d,J=8.6Hz,1H),5.58(q,J=7.9Hz,1H),4.61(q,J=7.7Hz,2H),3.53–3.43(m,1H) ,3.28–3.19(m,1H),3.06(s,3H),2.74–2.65(m,1H),1.94–1.84(m,1H).HRMS(ESI)m / z calcd for C 21 H 18 F3N4O4S[M+H] + 479.1001, found 479.1031

[0250] Example 16

[0251] Synthesis of N-((1S)-4-(5-(3-cyano-4-(2,2,2-trifluoroethoxy)phenyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)ethanesulfonamide (IS-20), the structural formula of which is as follows:

[0252]

[0253] Its synthetic route is as follows:

[0254]

[0255] The synthesis method is as follows:

[0256] Referring to the synthesis method of Example 13, compound IS-20 was prepared using IS-9 and ethanesulfonyl chloride as raw materials.

[0257] 1H NMR(400MHz,Chloroform-d)δ8.49(d,J=2.2Hz,1H),8.42(dd,J=8.9,2.2Hz,1H),8.09(d,J=7 .6Hz,1H),7.49(d,J=7.5Hz,1H),7.40(t,J=7.6Hz,1H),7.18(d,J=8.9Hz,1H),5.72(d,J=8.6H z,1H),5.58(q,J=7.9Hz,1H),4.61(q,J=7.7Hz,2H),3.52–3.44(m,1H),3.26–3.17(m,1H),3.0 9(q,J=8.5Hz,2H),2.74–2.65(m,1H),1.94–1.84(m,1H),1.26(t,J=8.9Hz,3H).HRMS(ESI)m / z calcd for C 22 H 20 F3N4O4S[M+H] + 493.1157, found 493.1142.

[0258] Embodiment 17

[0259] Synthesis of (S)-5-(3-(1-((2-methoxyethyl)amino)-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)-2-(2,2,2-trifluoroethoxy)benzonitrile (IS-17), the structural formula of which is as follows:

[0260]

[0261] Its synthetic route is as follows:

[0262]

[0263] The synthesis method is as follows:

[0264] The synthetic method was similar to that in Example 1 to prepare compound IS-17.

[0265] 1H NMR (400MHz, DMSO-d6) δ9.39(m,2H),8.59(d,J=2.2Hz,1H),8.47(dd,J=9.0,2.3Hz,1H ),8.15(d,J=7.7Hz,1H),8.01(d,J=7.6Hz,1H),7.67–7.53(m,2H),6.67–6.34(m,1H), 5.28(t,J=5.2Hz,1H),4.75–4.64(m,2H),3.73(q,J=5.2Hz,2H),3.35(s,3H),3.30–3. 21(m,1H),3.02(d,J=25.1Hz,2H),2.59–2.51(m,1H),2.36–2.26(m,1H).HRMS(ESI)m / z calcd for C 23 H 22 F3N4O3[M+H] + 459.1644, found 459.1640.

[0266] Embodiment 18

[0267] Synthesis of (S)-N-(4-(5-(3-cyano-4-(2,2,2-trifluoroethoxy)phenyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)-2-hydroxyacetamide (IS-18), the structural formula of which is as follows:

[0268]

[0269] Its synthetic route is as follows:

[0270]

[0271] Step 1: Synthesis of IS-18

[0272] IS-9 (0.3 g, 0.687 mmol) was added to 5 mL of dichloromethane, and triethylamine (0.069 g, 0.688 mmol), 2-hydroxyacetic acid (0.052 g, 0.688 mmol), 4-dimethylaminopyridine (0.017 g, 0.139 mmol) and N, N'-dicyclohexylcarbodiimide (0.15 g, 0.728 mmol) were added in sequence, and stirred at room temperature overnight. After the reaction was completed, the mixture was filtered and the filter cake was washed with dichloromethane. The filtrate was concentrated and the crude product was purified by column chromatography to obtain IS-18 (0.26 g, 82.5%).

[0273] 1H NMR(400MHz,Chloroform-d)δ8.49(d,J=2.2Hz,1H),8.42(dd,J=8.9,2.2Hz,1H),8.09(d,J =7.6Hz,1H),7.49(d,J=7.5Hz,1H),7.40(t,J=7.6Hz,1H),7.18(d,J=8.9Hz,1H),5.72(d,J= 8.6Hz,1H),5.58(q,J=7.9Hz,1H),4.61(q,J=7.7Hz,2H),4.21(d,J=5Hz,2H),3.92(br,1H) ,3.52–3.44(m,1H),3.26–3.17(m,1H),2.74–2.65(m,1H),1.94–1.84(m,1H).HRMS(ESI)m / z calcd for C 22 H 18 F3N4O4[M+H] + 459.1280, found 443.1301

[0274] Embodiment 19:

[0275] Synthesis of (S)-N-(4-(5-(3-cyano-4-(2,2,2-trifluoroethoxy)phenyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)-2-aminoacetamide (IS-19), the structural formula of which is as follows:

[0276]

[0277] Its synthetic route is as follows:

[0278]

[0279] IS-9 (0.3 g, 0.687 mmol) was added to 5 mL of dichloromethane, and triethylamine (0.069 g, 0.688 mmol), Boc-glycine (0.12 g, 0.688 mmol), 4-dimethylaminopyridine (0.017 g, 0.139 mmol) and N,N'-dicyclohexylcarbodiimide (0.15 g, 0.728 mmol) were added in sequence, and stirred at room temperature overnight. After the reaction was completed, the mixture was filtered and the filter cake was washed with dichloromethane. The filtrate was concentrated, 10 mL of 1M HCl in methanol was added, and the mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was concentrated, 20 mL of water was added to the residue, and the pH was adjusted to 9-10 with triethylamine. The mixture was extracted 3 times with ethyl acetate, and the organic phases were combined and washed 3 times with saturated brine. The organic phase was dried over anhydrous sodium sulfate and concentrated, and the crude product was purified by column chromatography to obtain IS-19 (0.17 g, 54.1%).

[0280] 1 H NMR(400MHz,Chloroform-d)δ8.49(d,J=2.0Hz,1H),8.42(dd,J=9.0,2.2Hz,1H),8.09(d,J =7.6Hz,1H),7.49(d,J=7.5Hz,1H),7.40(t,J=7.6Hz,1H),7.18(d,J=9.0Hz,1H),5.72(d,J =8.6Hz,1H),5.58(q,J=8.0Hz,1H),4.60(q,J=7.7Hz,2H),4.22(d,J=5Hz,2H),3.70(s,2H) ,3.52–3.44(m,1H),3.27–3.19(m,1H),2.74–2.65(m,1H),1.96–1.84(m,1H).HRMS(ESI)m / z calcd for C 22 H 19 F3N5O3[M+H] + 458.1440, found 458.1449.

[0281] Experimental Example 1: S1PR1 agonist activity detection

[0282] Ozamod used in this Experimental Example 1 was prepared with reference to the prior art (patent publication number CN105061350B), and the purity was 96.7% as determined by HPLC.

[0283] 1. Cell culture and solution preparation:

[0284] Cell line: Flpin-CHO-S1P1

[0285] F12K complete medium (commercially purchased): F12K (Hyclone) + 10% (v / v) fetal bovine serum (AUSGeneX) + 1× penicillin-streptomycin (Gibco), penicillin 100 IU / mL, streptomycin 100 μg / mL in F12K complete medium.

[0286] Working buffer: 1× Hanks' buffer (Gibco) + 20 mM HEPES (Gibco) + 0.1% (v / v) BSA (Perkin Elmer) + 500 μM 3-isobutyl-1-methylxanthine (Sigma)

[0287] 2. Activity detection:

[0288] a) Flpin-CHO-S1P1 cells were cultured with F12K complete medium and seeded into a 384-well plate at a density of 8000 cells / well;

[0289] b) using working buffer to prepare test compounds (compounds prepared in Examples 1 to 19) into test compound solutions with concentrations of 8000 nM, 2000 nM, 500 nM, 125 nM, 31.25 nM, 7.81 nM, 1.95 nM, 0.49 nM, 0.12 nM, and 0.03 nM, respectively;

[0290] c) Add 2.5 μL / well of the test compound solution to a 384-well plate and incubate at 37°C for 10 min;

[0291] d) Prepare 8 μM forskolin (Selleck) solution with working buffer, add 2.5 μL / well to a 384-well plate and incubate at 37°C for 30 min;

[0292] e) Dilute the Eu-cAMP tracer in the LANCE Ultra cAMP Detection Kit (Perkin Elmer) to 50 times with the detection buffer in the LANCE Ultra cAMP Detection Kit (Perkin Elmer) and add 10 μL to each well;

[0293] f) Dilute the Ulight-anti-cAMP in the LANCE Ultra cAMP Detection Kit (Perkin Elmer) to 150 times with the detection buffer in the LANCE Ultra cAMP Detection Kit (Perkin Elmer), and add 10 μL to each well;

[0294] h) incubate at room temperature for 1 h;

[0295] i) Envision 2105 microplate reader, excitation wavelength 340nM, emission wavelength 615nm and 665nm;

[0296] g) Graphpad drawing and calculation of EC 50 .

[0297] 3. Experimental results

[0298] The EC of the compounds prepared in Examples 1 to 19 for S1PR1 50 As shown in Table 1, the data results show that the compounds of the present invention have good agonist activity on S1PR1.

[0299] Table 1 EC values ​​of the compounds prepared in Examples 1 to 19 for S1PR1 50

[0300]

[0301] The present invention provides a modulator of sphingosine 1-phosphate receptor subtype 1 and its application ideas and methods. There are many methods and ways to implement the technical solution. The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the protection scope of the present invention. All components not specified in this embodiment can be implemented by existing technologies.

Claims

1. A compound having the formula IR or IS, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester, prodrug, homologue, hydrate or solvate thereof, in, R1 is a C1-C6 alkyl group substituted by halogen; R2 and R3 are independently selected from hydrogen, C1-C6 alkyl, C1-C6 acyl, C1-C6 alkylsulfonyl, wherein the C1-C6 alkyl, C1-C6 acyl, C1-C6 alkylsulfonyl are independently unsubstituted or substituted with hydroxyl, amino, carboxyl, C1-C4 alkoxy, -NR4R5, C1-C4 alkyl; R4 and R5 are independently selected from C1-C6 alkyl.

2. The compound according to claim 1, characterized in that R1 is a C1-C4 alkyl group substituted by fluorine; preferably, R1 is a difluoromethyl group, a difluoroethyl group or a trifluoroethyl group.

3. The compound according to claim 1, characterized in that R2 and R3 are independently selected from hydrogen, C1-C4 alkyl, C1-C4 acyl, C1-C4 alkylsulfonyl, wherein the C1-C4 alkyl, C1-C4 acyl, C1-C4 alkylsulfonyl are independently unsubstituted or substituted by hydroxyl, amino, carboxyl, C1-C4 alkoxy, -NR4R5, C1-C4 alkyl, and R4 and R5 are independently selected from C1-C4 alkyl; preferably, R2 and R3 are independently selected from hydrogen, C1-C4 alkyl, C1-C4 acyl, C1-C4 alkylsulfonyl, wherein the C1-C4 alkyl, C1-C4 acyl, C1-C4 alkylsulfonyl are independently unsubstituted or substituted by hydroxyl, amino, carboxyl, C1-C4 alkoxy, -NR4R5, and R4 and R5 are independently selected from methyl or ethyl; more preferably, R2 and R3 are independently selected from hydrogen, methyl, hydroxyethyl, acetyl, methylsulfonyl, ethylsulfonyl, 4. The compound according to claim 1, characterized in that The compound is any one of the following structures:

5. The compound according to claim 1, characterized in that The pharmaceutically acceptable salt is hydrochloride, hydrobromide, sulfate, phosphate, nitrate, acetate, maleate, fumarate, citrate, tartrate, methanesulfonate, p-toluenesulfonate or benzenesulfonate.

6. Use of the compound according to any one of claims 1 to 5 in the preparation of an activator or agonist of sphingosine 1-phosphate receptor subtype 1.

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