A chiral aryl propionic acid derivative and its pharmaceutical composition and use

Abstract

The present invention discloses a chiral arylpropionic acid derivative and a pharmaceutical composition and use thereof. The chiral arylpropionic acid derivative is shown in formula (I). The chiral arylpropionic acid derivative has antipyretic, analgesic and anti-inflammatory effects. The compound of the present invention has high activity and can be locally administered, which is expected to reduce the dosage and adverse reactions.

Description

[0001] This application is a divisional application of an application with an application date of April 18, 2023, application number 202310412212.X, and invention name “A chiral aromatic propionic acid derivative, its pharmaceutical composition and use”. Technical Field

[0002] The present invention relates to the field of pharmaceutical chemistry technology, but is not limited thereto, and in particular to a chiral arylpropionic acid derivative and a pharmaceutical composition and use thereof. Background Art

[0003] Loxoprofen Sodium, chemical name: 2-[4-(2-oxocyclopentane-1-ylmethyl)phenyl] propionate sodium, is the first propionic acid precursor non-steroidal anti-inflammatory drug (NSAIDs), developed by Japan Sankyo Co., Ltd. In July 1986, loxoprofen sodium tablets were launched in Japan under the trade name: Loxonin. It was launched in my country in 1999, and the original Chinese trademark name is "Lesong". The dosage forms of the marketed drugs are tablets, capsules, granules, patches, and gels. Among the various dosage forms of the drugs on the market, the API form selected is loxoprofen sodium dihydrate.

[0004]

[0005] Loxoprofen is a prodrug with weak activity. After oral administration, it needs to be metabolized by the liver and converted into active ingredients to exert its therapeutic effect. Loxoprofen is believed to be metabolized by carbonyl reductase in the skin and subcutaneous muscle tissue after topical administration. Loxoprofen has 2 chiral centers and 4 chiral isomers. Currently, loxoprofen sodium is marketed in the form of a racemate. There is no literature report on the difference in the efficacy of the four isomers of loxoprofen. After oral administration, loxoprofen is rapidly absorbed. The plasma concentration of loxoprofen and its metabolites reaches a peak after 30 to 50 minutes, and the plasma protein binding rates are 97% and 93%, respectively; a single topical administration of 1% loxoprofen (100 mg) transfers 10% of the dose into the body within 12 hours.

[0006] After metabolism, the carbonyl group of loxoprofen is reduced to a hydroxyl group, and the structure will contain three chiral centers and eight isomers, one of which is the trans-OH active metabolite. There is no literature report on the specific pharmacodynamic differences between the corresponding non-corresponding isomers of its in vivo active metabolites containing three chiral centers.

[0007] We conducted a detailed study on several of its isomers and developed a highly effective and fast-acting non-steroidal anti-inflammatory drug derivative through structural modification. Summary of the invention

[0008] The inventors have developed a chiral aryl propionic acid derivative, which has antipyretic, analgesic and anti-inflammatory effects. The compound of the invention has a better anti-rheumatoid arthritis effect, and is expected to reduce the dosage and adverse reactions. In addition, it is unexpectedly found that it is distributed more in the joints. Further studies have shown that it has better synovial fluid distribution and better therapeutic effect when applied externally to the skin, and can be used as a topically administered non-steroidal anti-inflammatory drug.

[0009] In one aspect, the present invention provides a chiral arylpropionic acid derivative, tautomer, solvate, or pharmaceutically acceptable salt thereof as shown in (I):

[0010]

[0011] In formula (I), R 1 is H, or is selected from one or more Substituted or unsubstituted

[0012] R 2 is H, or is selected from one or more Substituted or unsubstituted

[0013] And R 1 When it is hydrogen, R 2 Not hydrogen;

[0014] Y 2 O, or N(R 6 ), or S;

[0015] n 1 、n 2 、n 3 、n 4 、n 5 and n 6 each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10;

[0016] R 3 , R 4 and R 5 Each is independently H, C1-C20 hydrocarbon group, or C1-C20 alkylcarbonyl group, or R 3 and R 4 Together with the connected nitrogen atoms, they form a ring;

[0017] R 6 is H, or a C1-C20 hydrocarbon group;

[0018] A - Represents acceptable inorganic or organic negative ions.

[0019] In an embodiment of the present application, the hydrocarbon group includes an alkane group, an alkene group or an alkynyl group, and also includes a heterocyclic group, an aryl group or a heteroaryl group.

[0020] In an embodiment of the present application, the hydrocarbon group includes a linear, branched or cyclic hydrocarbon group.

[0021] In the embodiments of the present application, the C1-C20 hydrocarbon group refers to a saturated or unsaturated aliphatic hydrocarbon group containing 1 to 20 carbon atoms, including C1-C20 alkane groups, C2-C20 olefin groups, and C2-C20 alkynyl groups. For example, but not limited to: methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, vinyl, ethynyl, propenyl, propynyl, allyl, 2-methyl-2-butenyl, 2-butenyl (-CH 2 -CH=CH-CH 3 ), 3-butenyl (-CH 2 -CH 2 -CH=CH 2 ), 4-pentenyl (-CH 2 -CH 2 -CH 2 -CH=CH 2 ), 2-methyl-2-pentenyl (-CH 2 -C(CH 3 )=CH-CH 2 -CH 3 ), 5-hexenyl (-CH 2 CH 2 CH 2 CH=CH 2 ).

[0022] In the embodiments of the present application, the hydrocarbon group contains a C2-C20 heterocyclic group, and the heterocyclic group means that the heterocyclic ring includes at least one or more atoms selected from the following: oxygen, nitrogen and sulfur; including but not limited to: oxirane, aziridine, thiirane, azetidine, oxetanyl, thitanyl, 1,2-dithiyl, 1,3-dithiyl, pyrrolidinyl, dihydro-1H-pyrrolyl, dihydrofuranyl, tetrahydrofuranyl, dihydrothiophenyl, tetrahydrothiophenyl, imidazolidinyl, piperidinyl, piperazinyl, isoquinolinyl , tetrahydroisoquinolyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, hexahydrothiopyranyl, hexahydropyrimidinyl, oxazacyclohexanyl, thiazinyl, thioxanyl, homopiperazinyl, homopiperidinyl, azepanyl, oxepanyl, thiazole, oxazepanyl, diazepanyl, 1,4-diazepanyl, thiazepanyl, tetrahydrothiopyranyl, oxazolidinyl, thiazolidinyl, isothiazolidinyl, 1,1-dioxoisothiazolidinone, oxazolidinone and the like.

[0023] In an embodiment of the present application, the hydrocarbon group comprises an aromatic group, and the aromatic group includes but is not limited to benzene, naphthalene, anthracene, biphenyl, and the like.

[0024] In an embodiment of the present application, the hydrocarbon group comprises a heteroaryl group, including but not limited to: pyrimidine, furan, thiazole, thiophene, pyridine, pyrrole, and imidazole.

[0025] In an embodiment of the present application, the pharmaceutically acceptable salt refers to any form of the chiral arylpropionic acid derivative used according to the present invention, wherein it is in an ionic form or is charged and is coupled to a counterion (anion or cation) or is in solution;

[0026] In an embodiment of the present application, the pharmaceutically acceptable salt comprises a salt of the chiral arylpropionic acid derivative of the present invention and an anion, wherein the anion includes but is not limited to: fluoride ion, chloride ion, bromide ion, iodide ion, acetate ion, benzoate ion, citrate ion, tartrate ion, oxalate ion, malate ion, ascorbate ion, fumarate ion, etc.;

[0027] In the embodiment of the present application, the pharmaceutically acceptable salt includes the chiral aryl propionic acid derivative of the present invention as an inner salt, and the inner salt refers to a salt formed within the molecule when the molecule contains both a carboxyl group and an amine group, and the same molecule has both a positive charge and a negative charge;

[0028] In the embodiments of the present application, the pharmaceutically acceptable salt includes intermolecular salts of the chiral aromatic propionic acid derivatives of the present invention, and the intermolecular salts refer to salts formed between different molecules when the chiral aromatic propionic acid derivative molecules contain both carboxyl groups and amine groups.

[0029] In some embodiments, R 1 is hydrogen, R 2 For one or more Replaced

[0030] In some embodiments, R 2 is hydrogen, R 1 For one or more Replaced

[0031] In some embodiments, R 1 For one or more Replaced R 2 For one or more Replaced

[0032] In some embodiments, R 1 For one or more Replaced R 2 for

[0033] In some embodiments, R 1 for R 2 For one or more Replaced

[0034] In some embodiments, R 1 or R 2 When not hydrogen, its substituent It can replace hydrogen on terminal carbon or non-terminal carbon.

[0035] In some embodiments, R 3 and R 4 All are H;

[0036] In some embodiments, R 3 and R 4 are C1-C20 hydrocarbon groups or C1-C20 alkylcarbonyl groups; preferably, R 3 and R 4 are C1-C6 hydrocarbon groups or C1-C6 alkylcarbonyl groups; preferably, R 3 and R 4 All are C1-C6 alkyl;

[0037] In some embodiments, R 3 For H, R 4is a C1-C20 hydrocarbon group or a C1-C20 alkylcarbonyl group; preferably, R 4 are C1-C6 hydrocarbon groups; more preferably, R 4 is a C1-C6 alkyl group;

[0038] In some embodiments, R 4 For H, R 3 is a C1-C20 hydrocarbon group or a C1-C20 alkylcarbonyl group; preferably, R 3 are C1-C6 hydrocarbon groups; more preferably, R 3 is a C1-C6 alkyl group;

[0039] In some embodiments, R 3 and R 4 Together with the connected nitrogen atom, they form a ring, wherein the nitrogen atom and R3 and / or R4 can be connected by a common carbon-nitrogen bond or an amide bond; R 3 and R 4 The ring formed by the nitrogen atoms connected together can be a four-membered ring, a five-membered ring, a six-membered ring, or a seven-membered ring.

[0040] In some embodiments, R 3 , R 4 and R 5 All are H;

[0041] In some embodiments, R 3 and R 4 are C1-C20 hydrocarbon groups or C1-C20 alkylcarbonyl groups, R 5 is H; preferably, R 3 and R 4 are C1-C6 hydrocarbon groups or C1-C6 alkylcarbonyl groups, R 5 is H; preferably, R 3 and R 4 All are C1-C6 alkyl, R 5 is H;

[0042] In some embodiments, R 3 , R 4 and R 5 are C1-C20 hydrocarbon groups or C1-C20 alkylcarbonyl groups; preferably, R 3 , R 4 and R 5 are C1-C6 hydrocarbon groups; preferably, R 3 , R 4 and R 5 All are C1-C6 alkyl;

[0043] In some embodiments, R 3 and R 4 Together with the connected nitrogen atom, R5 are H or C1-C20 hydrocarbon groups, wherein the nitrogen atom and R3 and / or R4 can be connected by a common carbon-nitrogen bond or an amide bond; R 3 and R 4 The ring formed by the nitrogen atoms connected together can be a four-membered ring, a five-membered ring, a six-membered ring, or a seven-membered ring.

[0044] In some embodiments, Y 2 All O, or N(R 6 ), or S;

[0045] In some embodiments, Y 2 or N(R 6 ), or S.

[0046] In some embodiments, Y 2 or N(R 6 ), or O.

[0047] In some embodiments, Y 2 O, or N(R 6 ), or S.

[0048] In some embodiments, R 6 is H;

[0049] In some embodiments, R 6 is a C1-C20 hydrocarbon group; preferably, R 6 is a C1-C6 hydrocarbon group; more preferably, R 6 It is a C1-C6 alkyl group.

[0050] In some embodiments, n1, n2 and n3 are each independently 0, or 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or 10, preferably, n1, n2 and n3 are each independently 0, or 1, or 2, or 3, or 4;

[0051] In some embodiments, n1, n2, and n3 are all zero.

[0052] In some embodiments, n4, n5 and n6 are each independently 0, or 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or 10, preferably, n1, n2 and n3 are each independently 0, or 1, or 2, or 3, or 4;

[0053] In some embodiments, n4, n5, and n6 are all zero.

[0054] In some embodiments, A -Selected from halide, perhalate, nitrate, sulfate, hydrogen sulfide, sulfite, phosphate, hydrogen phosphate, C1-C8 alkyl acid, C1-C8 alkyl sulfonate, C1-C8 alkyl sulfate, C1-C8 aryl sulfonate;

[0055] In some more specific embodiments, A - Preferred are sulfate, phosphate, acetate, propionate, chloride and bromide.

[0056] In some embodiments, when R 1 is hydrogen, R 2 For one or more Replaced When, under appropriate conditions, the chiral aryl propionic acid derivatives of the present invention can form internal salts or intermolecular salts;

[0057] In some embodiments, when R 1 is hydrogen, R 2 For one or more Replaced When, under appropriate conditions, the chiral arylpropionic acid derivatives of the present invention can form salts with other anions, and the anions are as described above.

[0058] In some embodiments, R 2 is hydrogen, R 1 For one or more Replaced When, under appropriate conditions, the chiral aryl propionic acid derivatives of the present invention can form salts with other anions, and the anions are as described above;

[0059] In some embodiments, R 1 For one or more Replaced R 2 For one or more Replaced When, under appropriate conditions, the chiral arylpropionic acid derivatives of the present invention can form salts with other anions, and the anions are as described above.

[0060] In some embodiments, the chiral arylpropionic acid derivative provided by the present invention is selected from the following compounds:

[0061]

[0062] or a pharmaceutically acceptable salt.

[0063] In another aspect, the present invention provides a pharmaceutical composition comprising the chiral arylpropionic acid derivative, tautomer, solvate or a pharmaceutically acceptable salt thereof.

[0064] The invention discloses a pharmaceutical composition, which is composed of the compound, tautomer, solvate or pharmaceutically acceptable salt thereof as an active ingredient or a main active ingredient, and a pharmaceutically acceptable carrier.

[0065] The chiral arylpropionic acid derivatives of the present invention can be formulated into pharmaceutical compositions and administered to patients in a variety of suitable administration routes, including parenteral topical administration, such as skin topical preparations, eye preparations, inhalation preparations, etc.

[0066] In some embodiments of the present invention, the chiral arylpropionic acid derivative of the present invention and lactose are mixed and crushed to prepare an inhalant.

[0067] In some embodiments of the present invention, the chiral aryl propionic acid derivative of the present invention and a suitable amount of a surfactant and an osmotic pressure regulator are dissolved together to prepare a solution for inhalation.

[0068] In some embodiments of the present invention, the chiral arylpropionic acid derivative of the present invention and an appropriate amount of a surfactant are prepared together to prepare a skin external preparation.

[0069] In some embodiments of the present invention, the chiral arylpropionic acid derivatives of the present invention and suitable excipients are prepared into ophthalmic preparations.

[0070] Thirdly, the compounds of the present invention have high bioavailability when administered intra- or extra-intestinally and are rapidly converted into active ingredients, and thus have better effects when administered to the skin or orally.

[0071] The compound of the present invention has a better anti-rheumatoid arthritis effect. The compound of the present invention has a significant inhibitory effect on the proliferation of human rheumatoid arthritis fibroblast-like synoviocytes (HFLS-RA) induced by INF-α, and significantly inhibits the expression of inflammatory cytokines, showing higher activity, and is expected to reduce the dosage and adverse reactions.

[0072] In addition, it was unexpectedly found that the compound of the present invention has a higher distribution in the joints after oral administration. Further studies have shown that it has better joint fluid distribution and better therapeutic effect when applied externally to the skin, and can be used as a topically administered non-steroidal anti-inflammatory drug.

[0073] In the fourth aspect, the present invention provides the use of the above-mentioned chiral aromatic propionic acid derivatives, tautomers, and pharmaceutically acceptable salts thereof as non-steroidal anti-inflammatory drugs, which are mainly suitable for anti-inflammatory and analgesic effects for arthritis, rheumatoid arthritis, low back pain, scapulohumeral periarthritis, cervical shoulder-wrist syndrome and the like, as well as postoperative anti-inflammatory and analgesic effects and antipyretic and analgesic effects for acute respiratory inflammation. BRIEF DESCRIPTION OF THE DRAWINGS

[0074] Figure 1 It shows the comparative distribution of control 1, control 2 and the compounds of the present invention DSC4813 and DSC4821 in the joints. DETAILED DESCRIPTION

[0075] The following examples can enable those skilled in the art to more fully understand the present invention, but are not intended to limit the present invention in any way. The structures of all compounds are determined by MS or 1 H NMR confirmed.

[0076] Example 1: Synthesis of trans-OH metabolite

[0077] The compound trans-OH metabolite was synthesized with reference to the literature (Mandai, T. and T. Yamakawa (2000). "An Efficient Synthesis of (2S)-2-[4-((1R,2S)-2-Hydroxycyclopentylmethyl)phenyl]propionic Acid." Synlett 2000 (06): 0862-0864.), mp 87-88 ° C, [MH] - =247.16. 1 HNMR (300MHz, CDCl 3 )δ:1.20-1.32(m,1H),1.41-1.82(m,7H),1.88-2.06(m,2H),2.46(dd,1H),2.75(dd,1H),3.69(q,1H),3.86-3.95(m,1H),7.10-7.24(m,4H).

[0078] Example 2: Synthesis of DSC4801

[0079]

[0080] 0.8g of compound trans-OH metabolite was added to 10mL of N,N-dimethylformamide (DMF), the system was cooled to 0°C, 1.01g of compound 1 (2-bromo-N,N-diethylethylamine hydrobromide) and 0.85g of sodium carbonate were added, and the system was stirred for 18 hours. Water was added to precipitate the solid, which was filtered, and 20mL of dichloromethane was added to the solid, and water was added to separate the solid. The organic phase was concentrated to dryness and purified by silica gel column to obtain 0.72g of compound DSC4801, with a yield of 64%, [M+H] + =348.33, 1 H NMR (300 MHz, CDCl 3 )δ:0.95(t,6H),1.19-1.30(m,1H),1.41-1.71(m,6H),1.75-1.85(m,1H),1.94-2.14(m,2H),2.44( dd,1H),2.51(m,4H),2.78(m,3H),3.74(q,1H),3.86-4.05(m,1H),4.26(t,2H),7.11-7.21(m,4H).

[0081] Example 3: Synthesis of DSC4804

[0082]

[0083] 0.45 g of compound DSC4801, 0.15 g of pyridine and 10 mL of acetonitrile were added to the reaction flask, the reaction mixture was heated to 35°C, 0.2 g of acetic anhydride was added at 35-40°C, and then reacted at 50°C for 4 hours. The system was concentrated to dryness, cooled to room temperature, 15 mL of dichloromethane and 15 mL of water were added, the liquids were separated, and the organic phase was concentrated to dryness and purified by silica gel column to obtain 0.39 g of compound DSC4804, with a yield of 77%; [M+H] + =390.19, 1 H NMR (300 MHz, CDCl 3 )δ:1.08(t,6H),1.17-1.30(m,1H),1.45-1.86(m,7H),1.94-2.11(m,5H),2.49(dd,1H), 2.53(m,4H),2.75(m,3H),3.76(q,1H),3.81-3.98(m,1H),4.3(t,2H),7.07-7.20(m,4H).

[0084] Example 4: Synthesis of DSC4807

[0085]

[0086] Synthesis of compound 2

[0087] 1.0 g of compound trans-OHmetabolite was added to 10 mL of N,N-dimethylformamide (DMF), the system was cooled to 0°C, 0.83 g of benzyl bromide and 0.55 g of sodium carbonate were added, and the system was stirred for 3 hours. Water was added to precipitate the solid, which was filtered, and 20 mL of dichloromethane was added to the solid, and water was added to separate the solid. The organic phase was concentrated to dryness and purified by silica gel column to obtain 1.12 g of compound 2, with a yield of 82%; [M+H] + =339.10;

[0088] Synthesis of compound 3

[0089] 1.0 g of compound 2, 0.3 g of pyridine and 20 mL of anhydrous dichloromethane were added to the reaction flask, the reaction mixture was cooled to 0°C, 0.66 g of bromoacetyl bromide was added, the system was reacted at room temperature for 12 hours, water was added to separate, the organic phase was concentrated to dryness and purified by silica gel column to obtain 1.06 g of compound 3, the yield was 78%; [M+H] + =459.21;

[0090] Synthesis of compound 4

[0091] 0.2g of compound diethylaminoethanol and 15mL of anhydrous tetrahydrofuran were added to the reaction flask, the system was cooled to 0°C, 0.05g of sodium hydride (60%) was added, and the system was stirred for 30 minutes. 0.71g of compound 3 was added, the reaction mixture was heated to 35°C for 4 hours, the system was concentrated to dryness, cooled to room temperature, 15mL of dichloromethane and 15mL of water were added, the liquids were separated, the organic phase was concentrated to dryness and purified by silica gel column to obtain 0.63g of compound 4, with a yield of 75%; [M+H] + =496.34;

[0092] Synthesis of compound DSC4807

[0093] 1.19 g of 10% palladium on carbon was added to 0.5 g of compound 4, 10 mL of methanol was added to the system, a hydrogen balloon was connected, and the system was heated to 30°C for 8 hours. The system was filtered, the filtrate was concentrated to dryness, and purified by silica gel column to obtain 0.36 g of compound DSC4807, with a yield of 88%; [MH] - =404.29.

[0094] Example 5: Synthesis of DSC4806

[0095]

[0096] Synthesis of compound 6

[0097] Compound 6 was synthesized according to the method in the literature ("Characterization of N,N-dimethyl amino acids by electrospray ionization–tandemmass spectrometry." J. Mass Spectrom. 2015, 50, 771–781).

[0098] Synthesis of compound 7

[0099] 1.2 g of compound 6 and 30 mL of ethanol were added to the reaction flask, 10 mL of 2N sodium hydroxide solution was added to the system, and 1.51 g of benzyl bromide was added under vigorous stirring. After the system reacted at room temperature for 2 hours, concentrated hydrochloric acid was added to adjust the pH to neutral, and solids precipitated. The filter cake was washed with water and ethanol in turn, and the system was dried to obtain 1.48 g of compound 7, with a yield of 77%, [MH] - =238.25.

[0100] Synthesis of compound 8

[0101] 1.2 g of compound 7 and 15 mL of dichloromethane were added to the reaction flask, 1 g of dichlorothionyl was added to the system, and the reaction was carried out at room temperature for 3 hours. The system was concentrated to dryness, and 30 mL of dichloromethane was added again and concentrated to dryness. 15 mL of dichloromethane and 1 mL of triethylamine were added to the system, and 1.74 g of compound DSC4801 was added. The system was heated to 40 degrees and reacted for 5 hours, concentrated to dryness, and purified by silica gel column to obtain 1.83 g of compound 8, with a yield of 64%, [M+H] + =569.33;

[0102] Synthesis of compound DSC4806

[0103] 0.25 g of 5% palladium on carbon was added to 0.6 g of compound 8, 10 mL of methanol was added to the system, a hydrogen balloon was connected, and the system was heated to 30°C for 8 hours. The system was filtered, the filtrate was concentrated to dryness, and purified by silica gel column to obtain 0.43 g of compound DSC4806, with a yield of 85%; [M+H] + =479.29.

[0104] Example 6: Synthesis of DSC4815 and DSC4816

[0105]

[0106] Synthesis of compound DSC4815

[0107] Under nitrogen protection, at room temperature, 20 mL of tetrahydrofuran and 2.6 g of compound DSC4801 were added to the reaction bottle in sequence, and 8 mL of tetrahydrofuran solution of 0.54 g of ethyl chloride was slowly added. After the addition was completed, the mixture was heated to reflux and reacted for 30 minutes. After the reaction was completed, the temperature was lowered to 0-10°C, filtered, and dried to obtain a crude product, which was recrystallized from a methanol / acetone mixed solvent to obtain 1.33 g of compound DSC4815, with a yield of 43%, [M+H] + =376.03;

[0108] Synthesis of compound DSC4816

[0109] 1 g of compound DSC4815 and 5 mL of anhydrous ethanol were added to the reaction flask, heated at 50 °C, 0.45 g of silver acetate was added, the reaction mixture was stirred for 3 h, hot filtered, the filtrate was cooled to about 10 °C, 13 mL of methyl tert-butyl ether was added, and crystallization was performed to obtain 0.39 g of compound DSC4816, with a yield of 37%, [M+H] + =376.03,[MH] - =59.01.

[0110] Example 7: Synthesis of DSC4821

[0111]

[0112] Synthesis of compound 11

[0113] 0.18 g of diethylamine hydrochloride, 0.5 g of compound 3 and 15 mL of anhydrous tetrahydrofuran were added to the reaction flask, the system was cooled to 0 ° C, 0.7 mL of triethylamine was added, the reaction mixture was heated to 35 ° C for 4 hours, the system was concentrated to dryness, cooled to room temperature, 15 mL of dichloromethane and 15 mL of water were added, the liquids were separated, the organic phase was concentrated to dryness, and purified by silica gel column to obtain 0.34 g of compound 11, with a yield of 69%; [M+H] + =452.34;

[0114] Synthesis of compound DSC4821

[0115] 50 mg of palladium on carbon (10%) was added to 0.3 g of compound 11, 10 mL of methanol was added to the system, a hydrogen balloon was connected, and the system was heated to 30°C for 8 hours. The system was filtered, the filtrate was concentrated to dryness, and purified by silica gel column to obtain 0.2 g of compound DSC4821, with a yield of 84%; [MH] - =360.29, 1 H NMR (300 MHz, CDCl 3)δ:0.99(t,6H),1.22-1.30(m,1H),1.37-1.66(m,3H),1.45(d,3H),1.70-1.79(m,1H),1.83-2.02(m, 2H),2.43-2.66(m,6H),2.75(dd,1H),3.15(s,2H),3.65(q,1H),3.76-3.88(m,1H),7.08-7.27(m,4H).

[0116] Example 8: Synthesis of DSC4826

[0117]

[0118] Synthesis of compound 12

[0119] 0.7 g of compound 2, 1.25 g of p-nitrophenyl chloroformate, 1 mL of triethylamine and 10 mL of tetrahydrofuran were added to the reaction flask. The system was reacted at room temperature for 1 hour. The system was concentrated to dryness, ethyl acetate and water were added for extraction, and the organic phase was concentrated to dryness and purified by silica gel column to obtain 0.8 g of compound 12 with a yield of 77%, [M+H] + =504.21;

[0120] Synthesis of compound 13

[0121] 0.13g of compound diethylaminoethanol and 15mL of anhydrous tetrahydrofuran were added to the reaction flask, the system was cooled to 0°C, 65mg of sodium hydride (60%) was added, and the system was stirred for 30 minutes. 0.55g of compound 12 was added, the reaction mixture was heated to 35°C for 4 hours, the system was concentrated to dryness, cooled to room temperature, 15mL of dichloromethane and 15mL of water were added, the liquids were separated, the organic phase was concentrated to dryness and purified by silica gel column to obtain 0.37g of compound 13, with a yield of 71%; [M+H] + =482.39;

[0122] Synthesis of compound DSC4826

[0123] Referring to the synthesis method of compound DSC4821, 0.18 g of compound DSC4826 was synthesized using compound 13 as the raw material, with a yield of 84%, [MH] - =390.33, 1 H NMR (300 MHz, CDCl 3)δ:1.01(t,6H),1.22-1.30(m,1H),1.38-1.82(m,7H),1.91-2.06(m,2H),2.46(dd,1H),2.51-2.5 5(m,4H),2.75-2.88(m,3H),3.71(q,1H),3.86-3.91(m,1H),4.23-4.25(t,2H),7.05-7.20(m,4H).

[0124] The following example compounds were synthesized by the same method as in the above examples using commercially available compounds or intermediate compounds appropriately synthesized from commercially available compounds.

[0125]

[0126]

[0127] Synthesis of Comparative Example

[0128]

[0129] The synthetic method of the synthetic reference example compound of control 1 was used to obtain [M+H] - =317.37, 1 H NMR (300 MHz, CDCl 3 )δ:0.78(t,3H),1.06-1.29(m,3H),1.38-1.76(m,7H),1.81-2.02(m,2H),2.11(t,2 H),2.47(dd,1H),2.79(dd,1H),3.63(q,1H),3.85-3.95(m,1H),7.13-7.27(m,4H).

[0130] Control 2 was purchased from the market.

[0131] The synthesis of control 3 refers to the synthesis of the synthesis method in the embodiments of this patent and patent CN103705496B.

[0132] Example 9: Effect on the proliferation activity of HFLS-RA cells

[0133] Grouping and sample concentration: The experiment was divided into blank control group (2 groups in total, group 1 and group 2, group 1 was used for CCK-8 detection at 0h, group 2 was used for CCK-8 detection at 72h; both groups did not contain test sample and TNF-α), TNF-α group (only TNF-α was added without test sample, and the final concentration of TNF-α was 10ng / mL), test sample group (the test samples were DSC4801~DSC4826 and control 1~control 3, the structures of control 1~control 3 can be seen in the comparative example synthesis part, the final concentrations of each test sample and TNF-α incubated with cells were 100μg / mL and 10ng / mL, respectively) and positive control group (the final concentrations of methotrexate and TNF-α incubated with cells were 1μg / mL and 10ng / mL, respectively).

[0134] Co-culture: HFLS-RA cells in the logarithmic growth phase were taken and cultured at 3x10 4 The cells were inoculated at a density of 100 μL / ml in a 96-well culture plate. The inoculation volume in each well was 100 μL. After inoculation, the plate was incubated at 37°C with 5% CO. 2 After 24 hours of incubation in an incubator (cell adhesion and confluence reached about 25%), appropriate amounts of sample solutions were added according to the grouping and final sample concentration, with 3 replicate wells for each group. Before adding samples (0 h), CCK-8 detection was performed on group 1 of the blank control group, and CCK-8 detection was performed on the remaining groups after 72 hours of incubation in an incubator.

[0135] Detection and calculation: discard the culture medium, add 100 μL of culture medium containing 10% CCK-8 solution to each well, incubate at 37°C for 2 h, measure the absorbance (OD value) at 450 nm with an enzyme reader, and calculate the cell proliferation rate according to the following formula. The results are shown in Table 1.

[0136] Calculation formula: Cell proliferation rate = [OD (72h) -OD (0h) ] / OD (0h) x100%, where OD (0h) Indicates the OD value at 0h, OD (72h) Indicates the OD value at 72h.

[0137] Table 1: Proliferation rate of HFLS-RA cells after 72 hours of treatment

[0138] Group Cell proliferation rate Group Cell proliferation rate Group Cell proliferation rate Blank control group 61.32±2.01 TNF-α group 88.57±3.66 Positive control group 18.37±1.32 DSC4801 31.18±1.82 DSC4802 27.26±2.41 DSC4803 30.17±1.85 DSC4804 29.36±2.33 DSC4805 29.28±1.92 DSC4806 24.19±2.24 DSC4807 22.23±3.04 DSC4808 23.11±2.25 DSC4809 22.47±1.99 DSC4810 24.95±2.65 DSC4811 22.37±1.78 DSC4812 23.81±2.21 DSC4813 22.34±2.72 DSC4814 24.36±1.99 DSC4815 28.31±2.26 DSC4816 29.71±2.83 DSC4817 23.32±2.46 DSC4818 22.54±2.14 DSC4819 20.36±2.10 DSC4820 21.11±3.56 DSC4821 20.96±1.85 DSC4822 31.54±2.23 DSC4823 22.78±1.87 DSC4824 22.17±2.29 DSC4825 22.53±2.66 DSC4826 21.43±1.94 Comparison 1 39.33±3.46 Comparison 2 48.25±3.21 Comparison 3 44.31±4.15

[0139] As can be seen from Table 1, compared with the blank control group, the TNF-α group significantly promoted the proliferation of HFLS-RA cells (P<0.001); compared with the TNF-α group, the positive control group and the test sample (compounds DSC4801 to DSC4826 of the present invention and control 1 to control 3) groups could significantly inhibit the proliferation of HFLS-RA cells induced by TNF-α (P<0.001); compared with control group 1, control group 2 and control group 3, the level of inhibition of TNF-α-induced HFLS-RA cell proliferation by the compounds of the present invention was significantly higher (P<0.001). The results show that the compounds of the present invention can exert anti-inflammatory effects by inhibiting the proliferation of HFLS-RA cells, and are significantly better than control compounds 1, 2 and 3.

[0140] Example 10: Effect on HFLS-RA cytokine secretion

[0141] The experiment was divided into a blank control group (without test article or TNF-α), a TNF-α group (the final concentration of TNF-α incubated with cells was 10 ng / mL), and a test article group (a total of 8 groups, namely DSC4801 group, DSC4807 group, DSC4810 group, DSC4813 group, DSC4821 group, DSC4826 group, control group 1, and control group 3. The final concentrations of the test articles and TNF-α incubated with cells were 100 μg / mL and 10 ng / mL, respectively).

[0142] HFLS-RA cells in the logarithmic growth phase were taken and 5×10 4 The cells were inoculated at a density of 10 cells / ml in a 6-well culture plate with a volume of 200 μL per well. After inoculation, the plates were incubated at 37°C with 5% CO 2 The cells were cultured in an incubator for 2 h, and appropriate amounts of each sample solution were added according to the grouping and final sample concentration, with 3 replicate wells in each group. After 48 h of continuous culture, the cell supernatant was collected, and the ELISA kit was used to detect the content of IL-6 and IL-8 in the supernatant. The results are shown in Table 2.

[0143] Table 2: Effects of samples on the expression of cytokines IL-6 and IL-8 in HFLS-RA

[0144] Group IL-6 IL-8 Group IL-6 IL-8 Blank control group 44.51±0.58 107.35±3.01 TNF-α group 93.85±1.74 162.08±7.35 DSC4801 46.26±0.35 119.08±4.55 DSC4807 48.01±0.77 121.66±4.78 DSC4810 47.88±0.72 118.57±3.92 DSC4813 46.82±0.63 119.48±4.07 DSC4821 46.58±0.61 117.72±3.43 DSC4826 47.59±0.45 117.77±4.23 Comparison 1 67.45±2.68 131.21±6.88 Comparison 3 74.02±3.53 135.76±7.61

[0145] As can be seen from Table 2, compared with the blank control group, TNF-α can significantly induce the expression of IL-6 and IL-8 inflammatory factors in HFLS-RA cells (P<0.001); compared with the TNF-α group, control group 1 and control group 3, the compounds of the present invention can significantly inhibit the expression of IL-6 and IL-8 inflammatory factors in HFLS-RA cells induced by TNF-α (P<0.001). The results show that the compounds of the present invention can exert anti-inflammatory effects by inhibiting the expression of IL-6 and IL-8 inflammatory factors, and are significantly better than control compounds 1 and 3.

[0146] Example 11: Joint tissue distribution

[0147] 36 healthy male SD rats, weighing 200±20g, were raised at room temperature of 20-26℃, humidity of 40%-70%, and light-dark cycle of 12h / 12h. The rats were free to eat during the feeding process. After 3 days of adaptive feeding, the SD rats were randomly divided into 4 groups (A / B / C / D groups), with 9 rats in each group. Each group was randomly divided into 3 time groups, with 3 rats in each time group. Each group was orally gavaged with the suspension of the test article (the test article was suspended with 1% MS), among which group A was given 3mg / kg of DSC4813, group B was given 2.94mg / kg of DSC4821, group C was given 2.47mg / kg of control 1, and group D was given 2mg / kg of control 2 (loxoprofen). Three rats in each group were anesthetized and killed at 0.5h, 1h, and 2h after administration, and the left and right ankle joints of the hind legs were dissected, the skin at the joints was peeled off, and the tissues around the ankle joints (including the medial and lateral collateral ligaments, medial and lateral deltoid ligaments, fascia, tendons, joint synovium and other tissues) were weighed. The joint tissue samples were cut into pieces and ultrasonically extracted with 10mL of methanol for 20min. After the extraction, they were placed in EP tubes, sealed, and stored at -20℃ for testing. The analytes were detected by LC-MS / MS (groups A / B / C all detected the compound trans-OHmetabolite, group D detected loxoprofen and all isomers of the carbonyl group reduced to OH, and the results of group D were the sum of the concentrations of loxoprofen and all isomers of the carbonyl group reduced to OH). The results are shown in the figure. Figure 1 shown.

[0148] Depend on Figure 1 It can be seen that after oral gavage, the distribution of the compounds DSC4813 and DSC4821 in the joints was significantly increased (P<0.001) compared with the control 1 and the control 2. Therefore, it is inferred that the compounds DSC4813 and DSC4821 of the present invention have a strong anti-arthritis effect and are superior to the control compounds 1 and 2.

[0149] Although the present invention has been disclosed as above with preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art may make some modifications and improvements without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention shall be based on the definition of the claims.

Claims

1. A chiral arylpropionic acid derivative, tautomer, or pharmaceutically acceptable salt thereof as shown in (I): In formula (I), R1 is H, or R2 is selected from Replaced Y2 is O, or N(R6), or S; n1, n2, n3, n4, n5 and n6 are each independently 0, 1, 2, 3, 4; R3 and R4 are each independently C1-C6 alkyl; R5 is H, or C1-C6 alkyl; R6 is H, or C1-C6 alkyl; A - Represents acceptable inorganic or organic negative ions.

2. The chiral arylpropionic acid derivative, tautomer, or pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R1 is hydrogen.

3. The chiral arylpropionic acid derivative, tautomer, or pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R1 is ethyl.

4. The chiral arylpropionic acid derivative, tautomer, or pharmaceutically acceptable salt thereof as claimed in claim 1 or 2, wherein the pharmaceutically acceptable salt comprises a salt thereof with an anion, wherein the anion is a fluoride ion, a chloride ion, a bromide ion, an iodide ion, an acetate ion, a benzoate ion, a citrate ion, a tartrate ion, an oxalate ion, a malate ion, an ascorbate ion, or a fumarate ion.

5. The chiral arylpropionic acid derivative, tautomer, or pharmaceutically acceptable salt thereof as claimed in claim 4, wherein the pharmaceutically acceptable salt comprises an internal salt or an intermolecular salt.

6. The chiral arylpropionic acid derivative, tautomer, or pharmaceutically acceptable salt thereof according to claim 2, selected from the following compounds:

7. The chiral arylpropionic acid derivative, tautomer, or pharmaceutically acceptable salt thereof according to claim 3, selected from the following compounds:

8. A pharmaceutical composition comprising the chiral arylpropionic acid derivative, tautomer, or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7.

9. Use of the chiral arylpropionic acid derivative, tautomer, or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, or the pharmaceutical composition according to claim 8 in the preparation of antipyretic, analgesic, and anti-inflammatory drugs.

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