WO2020007342A1

WO2020007342A1 - Application of biphenyl phosphate compound as gpr84 antagonist

Info

Publication number: WO2020007342A1
Application number: PCT/CN2019/094702
Authority: WO
Inventors: 南发俊; 谢欣; 肖瑜峰; 张庆; 陈林海; 方友臣
Original assignee: 中国科学院上海药物研究所
Priority date: 2018-07-06
Filing date: 2019-07-04
Publication date: 2020-01-09
Also published as: CN110680823A; CN110680823B

Abstract

Provided is an application of a biphenyl phosphate compound, used for preparing a GPR84 antagonist, used as a GPR84 antagonist, or used for preparing a drug for treating a disease associated with high expression or excessively high excitability of the GPR84 receptor; the structure of said biphenyl phosphate compound is as shown by general formula I; the definitions of each substituent are as described in the description and the claims. Biphenyl phosphate compounds, as GPR84 receptor antagonists, are expected to become a new type of drug capable of treating inflammation-related diseases such as multiple sclerosis, inflammatory bowel disease, arthritis, hepatic fibrosis, and pulmonary fibrosis.

Description

Application of biphenyl phosphate compounds as GPR84 antagonists

Technical field

The present invention relates to the application of biphenyl phosphate ester compounds. As a ligand molecule of G protein-coupled receptor 84 (GPR84), it has GPR84 antagonistic activity and can competitively inhibit GPR84. Activation of this receptor by agonists can be used for the treatment of related diseases caused by high GPR84 expression or excessive excitability, such as multiple sclerosis, inflammatory bowel disease, liver fibrosis, pulmonary fibrosis, arthritis Wait.

Background technique

As a type of chemical catalyst widely used in asymmetric catalysis, biphenyl phosphate compounds have been successfully implemented since 2004 when Akiyama and Terada independently reported asymmetric Maninch reactions catalyzed by chiral phosphate compounds. Such reactions as Mannich reaction, Picter-Spengler reaction, aza-Diels-Alder reaction, Friedel-Crafts reaction, and high enantioselective catalysis of aza-Ene reaction.

GPR84 (G protein-coupled receptor 84) is a medium-chain fatty acid (C9-C14) receptor first discovered by Wittenberger and others in 2001. GPR84 is mainly expressed in bone marrow and peripheral blood leukocytes (including neutrophils). Cells, eosinophils, basophils) and adipocytes. In the presence of lipopolysaccharide (LPS), GPR84 expression in monocytes / macrophages is up-regulated, and medium-chain fatty acids can significantly up-regulate the expression of the IL-12p40 subunit in the macrophage cell line RAW264.7 via GPR84, and regulate Th1 Cellular immune response promotes inflammation and plays an important role in the occurrence of inflammatory diseases such as multiple sclerosis (MS), inflammatory bowel disease, and arthritis. In addition, the occurrence of metabolic diseases such as obesity and diabetes is closely related to chronic inflammation. When macrophages invade adipose tissue, they can promote the occurrence of inflammatory responses by secreting cytokines. At the same time, GPR84 expression in fat cells will increase, indicating that GPR84 is also involved Cross-regulation between fatty acid metabolism and the immune system.

Since GPR84 can promote inflammation, it plays an important role in the occurrence of inflammation-related diseases. Therefore, by inhibiting the activity of the GPR84 through a GPR84 antagonist, it can treat inflammation-related diseases, such as multiple sclerosis, inflammatory bowel disease, arthritis, and the like.

So far, reports of GPR84 antagonists are very scarce, only two patent reports of Galapagos, Belgium (WO 2013/092791; WO 2014/095798).

At present, there is no report on the biological activity of biphenyl phosphate compounds in the field of biological activity, especially GPR84.

Summary of the invention

The purpose of the present invention is to provide the use of a novel biphenyl phosphate ester compound.

According to a first aspect of the present invention, there is provided the use of a compound of the structure represented by Formula I or a pharmaceutically acceptable salt, the use is:

(i) for the preparation of GPR84 antagonists;

(ii) used as a GPR84 antagonist;

(iii) for the preparation of a medicament for the treatment of related diseases caused by high expression or high excitability of the GPR84 receptor,

The compound or pharmaceutically acceptable salt of the structure represented by Formula I is:

Where

Y is O or S; X is O or S;

M is H, or an ion of the following metals: Li, Na, K, Ca, Mg, Cu, Fe, Zn, Al, Mn, or a conjugate acid of the following bases: NH ₃ , arginine, betaine, caffeine , Choline, N, N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethyl Morpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydroxycobalamin, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, hydrazone, poly Amine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine;

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ are each independently hydrogen, substituted or unsubstituted C ₁ to C ₆ alkyl, substituted or unsubstituted C ₁ to C ₆ silyl, substituted or unsubstituted C ₃ to C ₆ cycloalkyl, substituted or unsubstituted C ₆ to C ₁₆ aryl, substituted or unsubstituted 3-20 membered heteroaryl; or R ₁ , R ₂ Any two of R ₃ , R ₄ and R ₄ form a substituted or unsubstituted C ₃ to C ₆ cyclic hydrocarbon, a substituted or unsubstituted C ₆ to C ₁₀ aromatic ring, or a substituted or unsubstituted 3 -12-membered heteroaryl ring; or any two of R ₅ , R ₆ , R ₇ , and R ₈ and the connected C to form a substituted or unsubstituted C ₃ ～ C ₆ cyclic hydrocarbon, substituted or unsubstituted C ₆ ~ C ₁₀ aromatic ring, or substituted or unsubstituted 3-12 membered heterocyclic ring;

L is unsubstituted, bonded, substituted or unsubstituted C ₁ to C ₆ alkylene, or between L and R ₄ and connected C, and L and R ₈ and connected C are independently substituted or unsubstituted C ₃ -C ₇ cyclic hydrocarbons;

The above-mentioned substitution means having one or more substituents selected from the group consisting of: C ₁ to C ₆ alkyl, C ₁ to C ₆ alkoxy, C ₁ to C ₆ alkylamino, C ₆ to C ₁₆ aryl, halogen , Hydroxyl, amino, -COOC ₁ to C ₆ alkyl, -COOH.

In another preferred example, R ₁ is hydrogen, substituted or unsubstituted C ₁ to C ₆ alkyl, substituted or unsubstituted C ₃ to C ₆ cycloalkyl, or substituted or unsubstituted C ₆ to C ₁₆ aromatic. Group, a substituted or unsubstituted 3-12 membered heterocyclic ring; or a substituted or unsubstituted C ₆ to C ₁₀ aromatic ring formed between R ₁ and R ₂ and an attached C,

In another preferred example, R ₅ is hydrogen, substituted or unsubstituted C ₁ to C ₆ alkyl, substituted or unsubstituted C ₃ to C ₆ cycloalkyl, substituted or unsubstituted C ₆ to C ₁₆ aromatic A substituted or unsubstituted 3-12 membered heterocyclic ring; a substituted or unsubstituted C ₆ to C ₁₀ aromatic ring is formed between R ₅ and R ₆ and the connected C,

In another preferred example, R ₁ and R ₅ are each independently hydrogen, substituted or unsubstituted C ₁ to C ₆ alkyl, substituted or unsubstituted C ₆ to C ₁₆ aryl, or substituted or unsubstituted 4 -6-membered heteroaryl, the substitution means having one or more substituents selected from the group consisting of phenyl, C ₁ to C ₄ alkoxy, C ₁ to C ₄ alkyl, fluorine, chlorine, bromine , -N (C ₁ to C ₄ alkyl) (C ₁ to C ₄ alkyl), C ₆ to C ₁₆ aryl, hydroxyl, amino, -COOC ₁ to C ₆ alkyl, -COOH.

In another preferred example, R ₃ and R ₄ form a substituted or unsubstituted C ₃ to C ₆ cyclic hydrocarbon, a substituted or unsubstituted C ₆ to C ₁₀ aromatic ring, or a substituted or unsubstituted C ₃ to R. 3-10 membered heterocyclic ring;

In another preferred example, R ₇ and R ₈ form a substituted or unsubstituted C ₃ to C ₆ cyclic hydrocarbon, a substituted or unsubstituted C ₆ to C ₁₀ aromatic ring, or a substituted or unsubstituted C and C ₈ . 3-10 membered heterocyclic ring;

In another preferred example, a benzene ring is formed between R ₁ and R ₂ and the connected C.

In another preferred example, a benzene ring is formed between R ₅ and R ₆ and the connected C.

In another preferred example, benzo C ₃ -C ₆ cycloalkyl or naphthyl is formed between R ₃ and R ₄ and the connected benzene ring.

In another preferred embodiment, R ₃ and R ₄ may be fused to be unsubstituted, substituted or unsubstituted C ₃ to C ₆ and cycloalkyl, substituted or unsubstituted C ₆ to C ₁₀ and aryl, substituted or unsubstituted. Substituted C ₃ to C ₁₀ heteroaryl.

In another preferred example, R ₇ and R ₈ are unsubstituted, substituted or unsubstituted C ₃ to C ₆ cycloalkyl, substituted or unsubstituted C ₆ to C ₁₀ aryl, substituted or unsubstituted And C ₃ to C ₁₀ heteroaryl.

In another preferred example, L is none, a bond, or a C ₁ -C ₄ alkylene group; or between L and R ₄ and the connected C, and L and R ₈ and the connected C are each independently and the ring is C ₃ -C ₇ cyclic hydrocarbon.

In another preferred example, L is none, a bond, or a C ₁ -C ₄ alkylene group; or between L and R ₄ and a connected benzene ring, and L and R ₈ and a connected benzene ring each independently form a benzene And C ₃ -C ₆ cycloalkyl.

In another preferred example, each of the above substitutions also has one or more substituents selected from the group consisting of: oxo (= O),-(C ₁ to C ₆ alkylene) COOC ₁ to C ₆ alkyl ,-(C ₁ to C ₆ alkylene) COOH,-(C ₁ to C ₆ alkylene) CONR'R ", wherein R 'and R" are each independently selected from: hydrogen, C ₁ to C ₆ alkane Group,-(C ₁ to C ₆ alkylene) -C ₆ to C ₂₀ aryl,-(C ₁ to C ₆ alkylene) -O- (C ₁ to C ₆ alkylene) -O- ( C ₁ to C ₆ alkylene) -NHBoc,-(C ₁ to C ₆ alkylene) -C ₆ to C ₂₀ heteroaryl,-(C ₁ to C ₆ alkylene) -NHCO-C ₆ to C ₂₀ heteroaryl, wherein C ₆ to C ₂₀ aryl and C ₆ to C ₂₀ heteroaryl are optionally substituted with a group selected from the group consisting of oxo (= O) and C1 to C6 alkylamino groups.

In another preferred example, the C1-C6 alkylamino group includes, but is not limited to, NHCH ₃ , N (CH ₃ ) (CH ₂ CH ₃ ), NHCH ₃ , NH (CH ₂ CH ₃ ), N (CH ₃ ) (CH ₂ CH ₂ CH ₃ ), N (CH ₂ CH ₃ ) (CH ₂ CH ₂ CH ₃ ), N (CH ₃ ) ₂ , N (CH ₂ CH ₃ ) ₂ or N (CH ₂ CH ₂ CH ₃ ) ₂ .

In another preferred example, the substituent on the substituted aryl group is selected from the following group:

In another preferred example, the compound is:

In another preferred example, each ring and substituent described in Formula I is independently a corresponding group in each specific compound described in the specification.

In another preferred example, the compound is a racemic compound or a chiral compound. In another preferred example, when the compound is a chiral compound, each chiral carbon is independently an R configuration or an S configuration.

In another preferred example, the disease is multiple sclerosis, inflammatory bowel disease, arthritis, pulmonary fibrosis or liver fibrosis.

According to a second aspect of the present invention, there is provided a method for treating a related disease caused by high expression of GPR84 receptor or excessive excitability, and administering a compound of the present invention or a pharmaceutically acceptable salt to a patient in need.

The biphenyl phosphate compounds of the present invention have previously been reported more as chiral catalyzed ligand molecules, and their reports on GPR84 biological activity testing are currently unavailable. Through in-depth research on these compounds, this was discovered for the first time creatively. A class of compounds has unexpectedly high antagonistic activity against GPR84. The result of this GPR84 antagonistic activity is the first such discovery report in the world to date, which is very innovative. It is expected to become a new drug that can treat inflammation-related diseases such as multiple sclerosis, inflammatory bowel disease, arthritis, liver fibrosis, and pulmonary fibrosis.

Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equal to those described can be used in the method of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

It should be understood that, within the scope of the present invention, the above technical features of the present invention and the technical features specifically described in the following (such as the embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, I will not repeat them here.

detailed description

After extensive and intensive research, the inventors of the present application discovered for the first time that biphenyl phosphate compounds have a high antagonistic activity on the GPR84 target and can competitively inhibit the receptors caused by GPR84 agonists. Activation, which can be used to prepare drugs for treating related diseases caused by high expression of GPR84 receptor or excessive excitability, including diseases such as multiple sclerosis, inflammatory bowel disease, arthritis, liver fibrosis, pulmonary fibrosis, etc. . Based on this, the present invention has been completed.

definition

In the present invention, C ₆ -C ₁₀ means having 6-10 carbon atoms, C ₃ -C ₆ means having 3-6 carbon atoms, and so on.

The integers 0-4 refer to 0, 1, 2, 3, 4; 6-10 carbon atoms refer to 6, 7, 8, 9, 10 carbon atoms, and so on.

A 3-10 member means a ring having 3-10 ring atoms.

In the present invention, unless otherwise stated, terms such as aromatic ring, cycloalkane ring, and alkyl group have the same meanings as those skilled in the art. For example, alkyl refers to a saturated linear or branched hydrocarbon group; for example, -CH ₃ or -CH (CH ₃ ) ₂ ; alkylene refers to a saturated hydrocarbon group that formsally removes the remaining two monovalent hydrogens, including but not It is limited to methylene (-CH _2- ), ethylene (-CH ₂ CH _2- ), and the like. Alkoxy refers to -O- (alkyl), including but not limited to -OCH ₃ , -OCH ₂ CH ₃ and the like. Cycloalkyl refers to a saturated cyclic hydrocarbon group, such as cyclohexyl. Heterocyclic refers to heterocycloalkyl or heteroaryl ring, heterocycloalkyl refers to a saturated cyclic hydrocarbon group containing at least one heteroatom (such as N, O, or S); heteroaryl ring or heteroaryl refers to containing at least one heteroatom A heteroatom aromatic ring. A cyclic hydrocarbon means a saturated, unsaturated cyclic hydrocarbon group. Aryl or aromatic ring refers to a full-carbon monocyclic or fused polycyclic ring having a conjugated π-electron system, and includes phenyl (Ph), naphthyl, fluorenyl, anthracenyl, and phenanthryl.

Unless otherwise stated, aryl (ring), heteroaryl (ring), cycloalkane ring, cyclic hydrocarbon, alkyl, alkylene, alkoxy, silyl, alkylamino, cycloalkyl, Heterocycloalkyl includes both substituted and unsubstituted. Possible substituents include, but are not limited to: C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₁ -C ₆ alkylamino, C ₂ -C ₁₀ alkynyl, C ₃ -C ₂₀ cycloalkyl, C ₃ -C ₂₀ cycloalkenyl, C ₁ -C ₂₀ heterocycloalkyl, C ₁ -C ₂₀ heterocycloalkenyl, C ₁ -C ₁₀ Alkoxy, C ₆ -C ₁₆ aryl, aryloxy, heteroaryl, heteroaryloxy, amino, hydroxyl, halogen, mercapto, cyano, nitro, carboxyl and carboxylate groups and the like.

GPR84 antagonist

The GPR84 antagonist provided by the present invention is a compound having the structure of Formula I:

The definition of each substituent is the same as above.

Most preferably, the compound of formula I described in the present invention is a compound prepared in the examples.

The present invention also provides pharmaceutically acceptable salts thereof, including salts obtained by reacting a compound of formula I with an inorganic base or an organic base compound.

Salts derived from inorganic bases include, but are not limited to: aluminum, ammonium, calcium, copper, iron, ferrous, lithium, magnesium, manganese, manganese, potassium, sodium, Zinc salt and so on. Particularly preferred are ammonium, calcium, magnesium, potassium, and sodium salts.

Salts obtained from pharmaceutically acceptable organic non-toxic bases, including but not limited to salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ions Exchange resins such as arginine, betaine, caffeine, choline, N, N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, aminoethanol , Ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydroxycobalamin, isopropylamine, lysine, methylglucosamine, Porphyrin, piperazine, piperidine, amidine, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.

Preparation

Compounds of formula I can be obtained by one of the following routes or by reference (Org. Lett, 2013, 15, 10, 2382-2385; Chem. Eur. J. 2009, 15, 6008-6014; J. Org. Chem, 1991, 56, 423 -425; J. Org. Chem, 1991, 56,423-425) to achieve:

Route one:

The reaction is carried out in pyridine; the reaction temperature is 60 ° C to 100 ° C; the reaction time is about 1 to 24 hours; after the reaction is completed, it is extracted with solvents such as AcOEt, Et ₂ O, CH ₂ Cl ₂ , CHCl ₃ , washed with saturated brine, and After drying, the solvent was removed under reduced pressure at low temperature. The target product was obtained by column chromatography of the concentrate, and the obtained product was confirmed by a method such as NMR.

The definition of each substituent is as described above.

Line two

S2 reacts with NaH and chloromethyl methyl ether in tetrahydrofuran to obtain S3 at room temperature, the reaction time is 4-12 hours; S3 reacts with n-butyl lithium and halogen element under tetrahydrofuran to obtain S4 and S5 (for S4, the halogen element is 1 -1.5 equivalents, halogen simple substance for S5 and 2.5-3.5 equivalents), the reaction temperature is 0 ℃ ～ 25 ℃, the reaction time is 4-12 hours; S4, S5 and boric acid compounds are in tetratriphenylphosphonium palladium, sodium carbonate The intermediate was refluxed under DME for 10-24 hours to obtain the intermediate, and the intermediate was deprotected by hydrochloric acid / THF to obtain S6; S6 was obtained by the method described in route one to obtain P2.

Line three:

S7 can be obtained according to the method described in line 2. After reacting S7 with n-butyllithium and a halogenated compound in THF, S4 can be obtained by deprotection of hydrochloric acid / THF, and S4 can be obtained by the method described in line 1.

Route four: into salt

The raw material P1 was dissolved in EA, and an alkali (a conjugate base of M, a hydroxide of M, or a carbonate of M) was added and washed twice. The aqueous layer was back-extracted with EA, the EA layer was concentrated, and the crude product was subjected to silica gel column chromatography. The product P2 is obtained.

M is a cation of the following metals: Li, Na, K, Ca, Mg, Cu, Fe, Zn, Al, Mn;

Conjugated acids of or below: NH ₃ , arginine, betaine, caffeine, choline, N, N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2- Dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydroxycobalamin, isopropylamine, lysine Acid, methylglucosamine, morpholine, piperazine, piperidine, hydrazone, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc .; others The definition of each substituent is as described above.

use

As an antagonist of GPR84, a compound of formula I can competitively inhibit the activation of the receptor caused by an agonist of GPR84, and can be used to prepare a drug for the treatment of related diseases caused by high expression of GPR84 receptor or excessive excitability. The diseases include multiple sclerosis, inflammatory bowel disease, arthritis, liver fibrosis, and pulmonary fibrosis.

The compounds of the invention can be formulated for use in pharmaceutical compositions. A pharmaceutical composition contains a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

"Pharmaceutically acceptable carrier", "pharmaceutically acceptable carrier" or "pharmaceutically acceptable carrier" means one or more compatible solid or liquid fillers or gel substances which are suitable for human use and There must be sufficient purity and low enough toxicity. "Compatibility" here means that each component in the composition can blend with the active ingredient of the present invention (the compound of formula I or a pharmaceutically acceptable salt thereof) and each other without significantly reducing the active ingredient Effect. Examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, Magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween

), Wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

The compounds and pharmaceutical compositions of the present invention can be in a variety of forms, and can be administered orally or parenterally in the form of injections, such as capsules, tablets, granules, solutions, powders, powders, or syrups. The compounds and pharmaceutical compositions may be present in a suitable solid or liquid carrier and in a suitable sterilizing device for injection or instillation. The above formulations can be prepared by conventional pharmaceutical methods.

The compounds and pharmaceutical compositions of the present invention are useful for clinical use in mammals, including humans and animals, and can be administered via the oral, nasal, or gastrointestinal tract. The most preferred route of administration is oral.

The above-mentioned features mentioned in the present invention, or the features mentioned in the embodiments can be arbitrarily combined. All features disclosed in the specification of this case can be used in combination with any composition form, and each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equal, or similar purpose. Unless otherwise stated, the disclosed features are only general examples of equal or similar features.

The present invention will be further described below with reference to specific embodiments. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods in the following examples are not marked with specific conditions, usually in accordance with conventional conditions such as Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer Suggested conditions. Unless stated otherwise, percentages and parts are percentages by weight and parts by weight.

In the following examples, NMR was measured by Mercury-VX 300M, Mercury-VX400M or AVANCE-III 500M instrument manufactured by Varian, NMR calibration: δH 7.26ppm (CDCl ₃ ), 2.50ppm (DMSO-d ₆ ), 3.15ppm (CD ₃ OD); reagents are mainly provided by Shanghai Chemical Reagent Company; TLC thin-layer chromatography silica gel plate is produced by Shandong Yantai Huiyou Silica Gel Development Co., Ltd., model HSGF 254; Chemical plant branch production, model zcx-11, 200-300 mesh.

Example 1

Preparation of compound XYF248B

The raw material G1 (100 mg, 0.54 mmol) was dissolved in dry pyridine (1.5 mL), and heavy distilled POCl ₃ (250 uL, 2.68 mmol) was added dropwise under the protection of N ₂ , and the reaction was performed at 60 ° C. for 5 h. The reaction solution was cooled to room temperature, diluted with water, adjusted to pH = 2 with 1N HCl, extracted with ethyl acetate, and back-extracted with the aqueous layer three times, and dried over anhydrous Na ₂ SO ₄ . Filtration, concentration and silica gel column chromatography (DCM / MeOH = 20/1 to 10/1) gave the target compound XYF248 (92 mg, 69%, white solid). ¹ H NMR (d ₆ -DMSO, 500 MHz): δ 7.63 (dd, J = 7.5, 1.5 Hz, 2H), 7.49 (t, J = 7.5 Hz, 2H), 7.38 (t, J = 7.5 Hz, 2H ), 7.25 (d, J = 8.0Hz, 2H)

The following compounds were synthesized in the same way:

Example 2

Preparation of compound XYF512S

Material G1 (4g, 13.97mmol) was dissolved in anhydrous THF (30mL), under N ₂ was added NaH (1.4g, 34.9mmol), 25 ℃ reaction was added MOMCl (2.65ml, 34.9mmol) after 1h, After 5h reaction at room temperature, water was added to quench the reaction, EA extraction was performed, and anhydrous Na ₂ SO _{4 was} dried. Filtration, concentration and silica gel column chromatography (PE / EA = 20/1) gave the target compound G2 (3.2 g, 61%, white solid). G2 (200mg, 0.57mmol) was dissolved in anhydrous THF, 0.95ml n-butyllithium in hexane (1.5mmol) was added at 0 ° C, and after reacting for 3 hours at room temperature, iodine (408mg, 1.6mmol) was added at -78 ° C. After reaction for 1 h, it was quenched with water, extracted with EA, and dried over anhydrous Na ₂ SO ₄ . Filtration, concentration and silica gel column chromatography (PE / EA = 20/1) gave the target compound G3 (230 mg, 65%, white solid). G3 (200 mg, 0.32 mmol) was dissolved in DME, and tetratriphenylphosphine palladium (20 mg, 10%), 0.85 ml of a 2M sodium carbonate solution, and 3-borate thiophene (143 mg, 1.12 mmol) were added. After replacement by nitrogen at reflux overnight, filtered, and spin dry product was dissolved in 10ml THF, was added 1.5ml of concentrated hydrochloric acid, was stirred 4 h at RT, extracted with EA, dried over anhydrous Na2SO _4. Filtration, concentration and silica gel column chromatography (PE / EA = 15/1) gave the target compound G3 (120 mg, 83%, white solid). G4 refers to the method in Example 1 to obtain XYF512S ¹ H NMR (d ₆ -DMSO, 500MHz): δ 8.38 (dd, J = 2.5, 1.0 Hz, 2H), 8.25 (s, 2H), 8.03 (d, J = 8.0Hz, 2H), 7.94 (dd, J = 5.0, 1.0Hz, 2H), 7.65-7.63 (m, 2H), 7.42 (t, J = 8.0Hz, 2H), 7.23 (td, J = 8.0, 1.0Hz, 2H), 7.00 (d, J = 9.0Hz, 2H)

The following compounds were synthesized in the same way

Example 3

Preparation of compound XYF432S

The raw material G1 was purchased from a reagent company. G1 (200 mg, 0.57 mmol) was dissolved in anhydrous THF, and 0.95 ml of a n-butyllithium hexane solution (1.5 mmol) was added at 0 ° C. After 3 hours of reaction at room temperature, the solution was added at -78 ° C 1-iodopropane (272mg, 1.6mmol), warmed to room temperature for 1h the reaction was quenched with water, extracted with EA, dried over anhydrous Na ₂ SO _4. Filtration and concentration, the product was dissolved in 10 ml of THF, 1.5 ml of concentrated hydrochloric acid was added, and after stirring at room temperature for 4 hours, EA was extracted and dried over anhydrous Na ₂ SO ₄ . Filtration, concentration and silica gel column chromatography (PE / EA = 15/1) gave the target compound G2 (98 mg, 46%, white solid). The final synthesis of XYF432S is the same as the method in Example 1. 1H NMR (d6-DMSO, 500MHz): δ7.92 (d, J = 8.0Hz, 2H), 7.84 (s, 2H), 7.37 (t, J = 7.5Hz, 2H), 7.19 (t, J = 8.0 Hz, 2H), 7.08 (d, J = 8.5Hz, 2H), 3.17-3.11 (m, 2H), 2.79-2.75 (m, 2H), 1.81-1-75 (m, 4H), 1.02 (t, J = 7.5Hz, 6H).

The following compounds were synthesized in the same way:

Example 4

Preparation of compound XYF813S

Compound G1 was synthesized according to the literature (US2013 / 204014). Compound XYF616S (110mg, 0.18mmol) was dissolved in 5mL of methanol solution, and an aqueous solution of LiOH (43mg, 1.8mmol) was added. The mixture was reacted at reflux for 4h, cooled to room temperature, and 1M hydrochloric acid solution was added. Adjust the pH to acidic, add ethyl acetate and deionized water, extract with ethyl acetate, and wash the organic phase with deionized water and saturated brine, dry the organic phase with anhydrous sodium sulfate, filter, spin dry, and then flash The crude intermediate was separated and purified by column chromatography. The crude intermediate (24 mg, 0.039 mmol) was dissolved in 1.5 mL of anhydrous DMF, and BOP (21 mg, 0.047 mmol) was added at 0 ° C. After stirring at 0 ° C for half an hour, the amino group was added. DMF solution of compound (13.5mg, 0.050mmol) and DIPEA (10mg, 0.078mmol), stirred at 0 ° C for 5h, added deionized water and ethyl acetate, extracted with ethyl acetate, and the organic phase was deionized water and saturated common salt, respectively. The organic phase was washed with water, dried over anhydrous sodium sulfate, and then filtered. The organic phase was separated and purified by flash column chromatography (DCM: MeOH = 10: 1) to obtain XYF813S, 25 mg white solid, molar yield: 85%.

¹ H NMR (d ₆ -DMSO, 500 MHz): δ 8.50 (t, J = 5.5 Hz, 1 H), 8.35 (d, J = 9.5 Hz, 1 H), 8.19-00 (m, 12 H), 7.99 (d , J = 8.0Hz, 1H), 7.96 (d, J = 7.5Hz, 1H), 7.89 (s, 1H), 7.72 (s, 1H), 7.50-7.45 (m, 4H), 7.42-7.38 (m, 3H), 7.19-7.16 (m, 1H), 6.99 (d, J = 8.5Hz, 1H), 6.89 (d, J = 9.0Hz, 1H), 6.80 (d, J = 8.5Hz, 1H), 5.03- 4.94 (m, 2H), 2.68-2.65 (m, 2H), 2.23-2.20 (m, 2H), 1.65-1.57 (m, 4H).

The following compounds were synthesized in the same way

Example 5

Experimental purpose

A GPR84 antagonistic activity test of the compounds of the present invention was performed.

2. Source of materials

A human-derived GPR84 cell line was obtained by transfecting a plasmid encoding the GPR84 and Gα16 proteins in a HEK293 cell line. The fluorescent dye Fluo-4AM was purchased from Invitrogen.

3. Test principle

Intracellular Ca ²⁺ is a very important second messenger of G protein coupled receptor signaling pathway. When GPR84 coupled with Gα ₁₆ protein and agonist are combined, the intracellular Ca ²⁺ concentration can be significantly increased. . Fluo-4 is a Ca ²⁺ ion-specific fluorescent probe that can quantitatively bind to Ca ²⁺ ions and emit fluorescence. Therefore, the fluorescence detection method is used to detect the agonistic or antagonistic activity of the compound in a 96-well or 384-well flat-bottomed microtiter plate.

Detection of the inhibitory effect of GPR84 antagonists on receptors: After GPR84 cells are incubated with the fluorescent dye Fluo-4, they are added with different concentrations of antagonist compounds and incubated for a period of time to occupy the binding site (antagonistic binding site) of the agonist to GPR84. A certain concentration of agonist (6-OAU) is added to compete with the antagonist for binding sites. At the same time, it is excited by a light source with a wavelength of 485nm and the change of the fluorescence intensity of the dye caused by the change of the calcium ion concentration in the cell is detected. PRISM software calculated half-maximum inhibition concentration (IC _50).

4. Experimental process

Formulated HBSS: 0.4g / L KCl (5.4mM), 0.12g / L Na ₂ HPO ₄ · 12H ₂ O (0.3mM), 0.06g / L KH ₂ PO ₄ (0.4mM), 0.35g / L NaHCO ₃ ( 4.2mM), 0.14g / L CaCl ₂ (1.3mM), 0.10g / L MgCl ₂ · 6H ₂ O (0.5mM), 0.05g / L MgSO ₄ (0.6mM), 8.0g / L NaCl (137mM), Weigh out the above components and dissolve them with ultrapure water, adjust the pH to 7.4 with hydrochloric acid or NaOH solution, filter, and store at 4 ° C for one month.

Preparation of calcium buffer solution: first prepare 560mM D-glucose (100X) water-soluble storage solution, 250mM 1,2-diphenyl-4- (2-phenylsulfinyl) ethyl-3,5-pyrazolidinedione ( 1000X) storage solution. Then add 100 mL of HBSS to BSA (0.5 g), 560 mM D-glucose stock solution (1 mL) and 250 mM 1,2-diphenyl-4- (2-phenylsulfinyl) ethyl-3,5-pyridine Zolanedione stock solution (100 μL), to a final concentration of 0.5% BSA, 5.6 mM D-glucose, 250 μM, 1,2-diphenyl-4- (2-phenylsulfinyl) ethyl-3,5 -Pyrazolidinedione, mix well, ready to use.

Formulation of dyes: First, prepare 3% Cremophor EL (100X) PBS-dissolved storage solution and 2mM Fluo-4 (1000X) DMSO-dissolved storage solution, and then prepare 1 μL of 2mM Fluo-4AM and 10 μL of each dye per ml. Mix 3% Cremophor EL, then dilute with 1mL calcium buffer and mix.

The cells were seeded at a density of 40,000 cells / well onto a 96-well cell culture plate, and the culture was continued for more than 24 hours to make the cell density to 80-90% for experimental detection. Aspirate the culture medium in the wells of the cells to be tested, add 40 μL / well of freshly prepared dye, and place in a 37 ° incubator at a constant temperature for 40 to 50 minutes.

Formulate compounds during cell incubation (this step can also be prepared in advance): freshly prepare calcium buffer before the experiment to dilute the compound used as an antagonist to 1.5 times the final working concentration, and dilute the compound used as an agonist to the final work 3 times the concentration (if the compound is dissolved in DMSO, the DMSO concentration should not exceed 1% in the final work).

After the incubation step is completed, the dye is sucked up and discarded. After washing with calcium buffer solution, change to 50 μL of calcium buffer solution containing different concentrations of antagonists and incubate for another 10 min.

Use a FlexStation III microplate detector to add 25 μL / well of calcium buffer containing a certain concentration (generally an effective concentration of the agonist EC ₈₀ or so) to the stimulus. At the same time, stimulate with a 485nm light source and detect cells at the 525nm band. Changes in the fluorescence intensity of the dye caused by changes in the internal calcium ion concentration.

5. Experimental results

Table 1. IC _{50 of} GPR84 calcium flow model compounds

All documents mentioned in the present invention are incorporated by reference in this application, as if each document was individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims

A use of a compound of the structure represented by Formula I or a pharmaceutically acceptable salt, wherein the use is:

(i) for the preparation of GPR84 antagonists;

(ii) used as a GPR84 antagonist;

(iii) for the preparation of a medicament for the treatment of related diseases caused by high expression or high excitability of the GPR84 receptor,

The compound or pharmaceutically acceptable salt of the structure represented by Formula I is:

In the formula, Y is O or S; X is O or S;

M is H, or an ion of the following metals: Li, Na, K, Ca, Mg, Cu, Fe, Zn, Al, Mn, or a conjugate acid of the following bases: NH 3 , arginine, betaine, caffeine , Choline, N, N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethyl Morpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydroxycobalamin, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, hydrazone, poly Amine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine;

R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 are each independently hydrogen, substituted or unsubstituted C 1 to C 6 alkyl, substituted or unsubstituted C 1 to C 6 silyl, substituted or unsubstituted C 3 to C 6 cycloalkyl, substituted or unsubstituted C 6 to C 16 aryl, substituted or unsubstituted 3-20 membered heteroaryl; or R 1 , R 2 Any two of R 3 , R 4 and R 4 form a substituted or unsubstituted C 3 to C 6 cyclic hydrocarbon, a substituted or unsubstituted C 6 to C 10 aromatic ring, or a substituted or unsubstituted 3 -12-membered heteroaryl ring; or any two of R 5 , R 6 , R 7 , and R 8 and the connected C to form a substituted or unsubstituted C 3 ～ C 6 cyclic hydrocarbon, substituted or unsubstituted C 6 ~ C 10 aromatic ring, or substituted or unsubstituted 3-12 membered heterocyclic ring;

L is unsubstituted, bonded, substituted or unsubstituted C 1 to C 6 alkylene, or between L and R 4 and connected C, and L and R 8 and connected C are independently substituted or unsubstituted C 3 -C 7 cyclic hydrocarbons;

The above-mentioned substitution means having one or more substituents selected from the group consisting of: C 1 to C 6 alkyl, C 1 to C 6 alkoxy, C 1 to C 6 alkylamino, C 6 to C 16 aryl, halogen , Hydroxyl, amino, -COOC 1 to C 6 alkyl, -COOH.
The use according to claim 1, wherein R 1 is hydrogen, substituted or unsubstituted C 1 to C 6 alkyl, substituted or unsubstituted C 3 to C 6 cycloalkyl, substituted or unsubstituted C 6 to C 16 aryl, substituted or unsubstituted 3-12 membered heterocyclic ring; or

A substituted or unsubstituted C 6 to C 10 aromatic ring is formed between R 1 and R 2 and the connected C,

The above-mentioned substitution means having one or more substituents selected from the group consisting of: C 1 to C 6 alkyl, C 1 to C 6 alkoxy, C 1 to C 6 alkylamino, C 6 to C 16 aryl, halogen , Hydroxyl, amino, -COOC 1 to C 6 alkyl, -COOH.
The use according to claim 1, wherein R 5 is hydrogen, substituted or unsubstituted C 1 to C 6 alkyl, substituted or unsubstituted C 3 to C 6 cycloalkyl, substituted or unsubstituted C 6 to C 16 aryl, substituted or unsubstituted 3-12 membered heterocyclic ring; or

R 5 and R 6 form a substituted or unsubstituted C 6 to C 10 aromatic ring between C and C,

The above-mentioned substitution means having one or more substituents selected from the group consisting of: C 1 to C 6 alkyl, C 1 to C 6 alkoxy, C 1 to C 6 alkylamino, C 6 to C 16 aryl, halogen , Hydroxyl, amino, -COOC 1 to C 6 alkyl, -COOH.
The use according to claim 1, characterized in that R 3 and R 4 form a substituted or unsubstituted C 3 to C 6 cyclic hydrocarbon, and a substituted or unsubstituted C 6 to C 10 aromatic ring between C and C. Or a substituted or unsubstituted 3-10 membered heterocyclic ring;

The above-mentioned substitution means having one or more substituents selected from the group consisting of: C 1 to C 6 alkyl, C 1 to C 6 alkoxy, C 1 to C 6 alkylamino, C 6 to C 16 aryl, halogen , Hydroxyl, amino, -COOC 1 to C 6 alkyl, -COOH.
The use according to claim 1, wherein R 7 and R 8 form a substituted or unsubstituted C 3 to C 6 cyclic hydrocarbon, a substituted or unsubstituted C 6 to C 10 aromatic ring between C and R. Or a substituted or unsubstituted 3-10 membered heterocyclic ring;

The above-mentioned substitution refers to having one or more substituents selected from the group consisting of C 1 to C 6 alkyl, C 1 to C 6 alkoxy, C 1 to C 6 alkylamino, and C 6 to C 16 aryl. , Halogen, hydroxyl, amino, -COOC 1 to C 6 alkyl, -COOH.
The use according to claim 1, wherein L is none, a bond, or a C 1 -C 4 alkylene group; or between L and R 4 and a connected C, and between L and R 8 and a connected C Each independently and cyclically is a C 3 -C 7 cyclic hydrocarbon.
The use according to claim 1, wherein the substitution further has one or more substituents selected from the group consisting of oxo (= O),-(C 1 to C 6 alkylene) COOC 1 to C 6 alkyl,-(C 1 to C 6 alkylene) COOH,-(C 1 to C 6 alkylene) CONR'R ", wherein R 'and R" are each independently selected from: hydrogen, C 1 to C 6 alkyl,-(C 1 to C 6 alkylene) -C 6 to C 20 aryl,-(C 1 to C 6 alkylene) -O- (C 1 to C 6 alkylene Group) -O- (C 1 to C 6 alkylene) -NHBoc,-(C 1 to C 6 alkylene) -C 6 to C 20 heteroaryl,-(C 1 to C 6 alkylene) -NHCO-C 6 to C 20 heteroaryl, wherein C 6 to C 20 aryl and C 6 to C 20 heteroaryl are optionally substituted with a group selected from the group consisting of oxo (= O), C1 ~ C6 alkylamino.
The use according to claim 1, wherein the compound is:
The use according to claim 1, wherein the compound is a racemic compound or a chiral compound.
The use according to claim 9, characterized in that when the compound is a chiral compound, each chiral carbon is independently an R configuration or an S configuration.
The use according to claim 1, wherein the disease is multiple sclerosis, inflammatory bowel disease, arthritis, pulmonary fibrosis or liver fibrosis.