WO2008140251A2

WO2008140251A2 - Cyclooxygenase-2 inhibitors

Info

Publication number: WO2008140251A2
Application number: PCT/KR2008/002677
Authority: WO
Inventors: Jae Yeol Lee; Dong Joon Choo; Kyung Tae Lee; Jong Taik Moon; Ji Young Jeon
Original assignee: Kyung Hee University
Current assignee: Kyung Hee University
Priority date: 2007-05-14
Filing date: 2008-05-14
Publication date: 2008-11-20
Anticipated expiration: 2009-11-14
Also published as: WO2008140251A3

Abstract

The present invention relates to lH-pyrrole or furan-2,5dione derivatives and a pharmaceutical composition for inhibiting cyclooxygenase-2(COX-2), more specifically for treating or preventing inflammation, an allergic disorder, inflammatory dermatosis, an immunologic disorder, a neurodegenerative disorder, arthritis, pain or fever comprising the same.

Description

Cyclooxygenase-2 Inhibitors Technical Field

[1] The present invention relates to lH-pyrrole or furan-2,5dione derivatives and a pharmaceutical composition for inhibiting cyclooxygenase-2(COX-2), more specifically for treating or preventing inflammation, an allergic disorder, inflammatory dermatosis, an immunologic disorder, a neurodegenerative disorder, arthritis, pain or fever comprising the same.

[2]

Background Art

[3] A human body produces cyclooxygenase- 1 (COX-I) and cyclooxygenase-2(COX-2) as hormones which are associated with inflammation process. Cyclooxygenase- 1 is also called prostaglandin Ηjsynthase and expressed in most of tissues. Prostaglandin Ηjproduced by cyclooxygenase- 1 is involved in the synthesis and supply of arachidonic acid metabolites to regulate physiological processes such as protection of gastrointestinal cells, kidney function and platelet aggregation. Meanwhile, cy- clooxygenase-2 is also known as prostaglandin Η synthase, which produces prostaglandin Η to induce inflammation and pain. Cyclooxygenase-2 is expressed only

2 by stimulation differently from cyclooxygenase- 1.

[4] Non-steroidal anti-inflammation drugs (NSAIDs) such as aspirin and ibuprofen exhibit their medicinal effects through inhibiting the activity of cyclooxygenases and suppressing the biosynthesis of prostaglandins. However, the non-steroidal anti- inflammation drugs not only inhibit the activity of cyclooxygenase-2 to reduce inflammation but also inhibit the activity of cyclooxygenase- 1 to cause adverse effects such as renal toxicity and gastrointestinal ulceration. Therefore, there has been a need in the art for the development of an anti-inflammation drug selectively inhibiting cyclooxygenase-2 over cyclooxygenase- 1.

[5] Cyclooxygenase-2 inhibitors which have been commercialized until now include celecoxib (Celebrex) and valdecoxib (Bextra) developed by Pfizer, and rofecoxib (Vioxx) developed by Merck. Celecoxib (Celebrex) (a) can be used for the treatment of arthritis and severe pain, and rofecoxib (Vioxx) (b) can be used for the treatment of pain and inflammation. Meanwhile, valdecoxib (Bextra) (c) can be extensively used for the treatment of rheumatism and arthritis.

(a) Celecoxib(celebrex) (b) Rofecoxιb(vιoxx) (c) Valdecoxib(bextra)

[7] However, rofecoxib (Vioxx) of Merck was withdrawn from the market due to side effects on cardiovascular system in 2004, and celecoxib (Celebrex) and valdecoxib (Bextra) of Pfizer are being restrictively prescribed since they may increase the risk of cardioplegia and stroke. Therefore, there is still a need in the art for the development of a new chemical drug which can substitute the above drugs.

[8]

Disclosure of Invention Technical Problem

[9] The present inventors have extensively studied to develop a novel cyclooxygenase-2 selective inhibitor which is more effective but less toxic. As a result, they discovered that a compound of the following formula (I) has excellent cyclooxygenase-2 inhibitory effect.

[10] An object of the present invention is, therefore, to provide a compound of the following formula (I) or pharmaceutically acceptable salt thereof having excellent cyclooxygenase-2 inhibitory effect.

[H] Another object of the present invention is to provide a pharmaceutical composition for inhibiting cyclooxygenase-2 comprising the compound of the following formula (I) or pharmaceutically acceptable salt thereof as an active ingredient.

[12]

Technical Solution [13] One aspect of the present invention relates to a compound of the following formula (I) or pharmaceutically acceptable salt thereof having excellent cyclooxygenase-2 inhibitory effect.

(I) [15] wherein,

[16] R , R and R are each independently hydrogen, hydroxy, C -C lower alkoxy, R

C(=O)O, R S, R S(=O), R S(=O) , NH S(=O) ,R C(=O)NH, or (R C(=O)) N; or R

5 5 5 2 2 2 5 5 2 2 and R taken together with the carbon atoms to which they are attached form a heterocycle; [17] R is hydrogen, R S, R S(=O) or R S(=O) ;

4 5 5 5 2

[18] R is C -C lower alkyl; and

5 1 5

[19] X is O or NH.

[20] In the above formula (I), R , R and R are, preferably, each independently hydrogen, hydroxy, C -C lower alkoxy, R C(=O)O, R S, R S(=O) , NH S(=O) , or R C(=O)NH;

1 5 5 5 5 2 2 2 5 the heterocycle is, preferably, dioxolane or dioxane; and R is, most preferably, methyl. [21] [22] The term "C -C lower alkoxy" as used herein means a straight or branched alkoxy group having 1 to 5 carbon atoms, which includes methoxy, ethoxy, propanoxy, etc. but is not limited thereto. [23] The term "Heterocycle" as used herein means a 4 to 74nembered ring having 1 to 3 heteroatom(s) selected from the group consisting of oxygen, sulfur and nitrogen, which includes dioxolane, dioxane, etc. but is not limited thereto.

[24] The term "C -C lower alkyl" as used herein means a straight or branched hydrocarbon having 1 to 5 carbon atoms, which includes methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. but is not limited thereto. [25] The most preferable compound among the compounds of the present invention is selected from the following group.

[26] 3-(3-methoxyphenyl)-4-phenyl-l H-pyrrole-2,5dione;

[27] 3-benzo[ 1 ,3]dioxol-5-yl-4-phenylfuran-2,5dione;

[28] 3-benzo[l,3]dioxol-5-yl-4-phenyl-l H-pyrrole-2,5dione;

[29] 3-(3,4-dihydroxyphenyl)-4-phenyl-l H-pyrrole-2,5dione;

[30] 3-(3,4,5-trihydroxyphenyl)-4-phenyl-l H-pyrrole-2,5dione;

[31] 3-(4-thiomethoxyphenyl)-4-phenylfuran-2,5dione;

[32] 3-(4-thiomethoxyphenyl)-4-phenyl- lH-pyrrole-2,5dione;

[33] 3-(4-methanesulfonylphenyl)-4-phenyl- 1 H-pyrrole-2,5dione;

[34] 3-(4-aminosulfonylphenyl)-4-phenyl- 1 H-pyrrole-2,5dione;

[35] 3-(4-acetoxyphenyl)-4-(4-methanesulfonylphenyl)furan-2,5dione;

[36] 3-(4-hydroxyphenyl)-4-(4-thiomethoxyphenyl)- lH-pyrrole-2,5dione; and

[37] 3-(4-acetaminophenyl)-4-(4-thiomethoxyphenyl)furan-2,5dione. [38] [39] The compounds of the present invention can be prepared according to the processes depicted in the following Reaction Schemes 1, 2 and 3. However, those illustrated in the following Reaction Schemes represent only typical processes used in the present invention. The manipulation order, reagents, reaction conditions, etc. may be changed without limit.

[4⁾] [Reaction Scheme 1]

[42] As shown in the above Reaction Scheme 1, a furan-2,5dione derivative(3) according to the present invention can be prepared by reacting phenylgly oxalic acid (1) and phenylacetic acid (2) in acetic anhydride. The acetic anhydride is used as a reactant as well as solvent. The reaction temperature is preferably from room temperature to 13O⁰C, most preferably about 100⁰C. The reaction time is preferably 2-10 hours, most preferably about 3 hours. As a catalyst, an acid catalyst can be used, which includes titanium chloride(TiCl ), BF (OEt) , thionyl chloride (SOCl ), etc. but is not limited

4 3 2 2 thereto.

[43] Meanwhile, a lH-pyrrole-2,5dione derivative(4) according to the present invention can be prepared by treating a furan-2,5dione derivative (3) with methanol, dimethyl- formamide (DMF) and hexamethyldisilazane (HMDS). The reaction temperature is most preferably room temperature, and the reaction time is preferably 10-36 hours, most preferably 12 hours. Alternatively, ammonia gas can be used in a solvent such as acetonitrile and lower alcohol including methanol.

[44] A lH-pyrrole-2,5dione derivative(5) wherein R , R and/or R are hydroxy can be prepared by dealkylating a lH-pyrrole-2,5dione derivative(4) wherein R , R and/or R are alkoxy or taken together with the carbon atoms to which they are attached form dioxolane or dioxane. The dealkylating agent includes boron tribromide (BBr ), trimethylsilane iodide (TMSI), aluminium chloride (AlCl ), etc. but is not limited thereto.

[45] [Reaction Scheme 2]

2

[47] As shown in the above Reaction Scheme 2, a lH-pyrrole-2,5dione derivative (4) according to the present invention can be prepared by coverting phenylacetic acid (2) to acyl chloride, and then treating the acyl chloride with ammonia water to give pheny- lacetamide (6), followed by the condensation of the phenylacetamide (6) with an ester compound (7).

[48] [49] Meanwhile, the phenylglyoxalic acid (1) and ester compound (7) used in the above Reaction Schemes 1 and 2 can be prepared according to a known process as shown in the following Reaction Scheme 3.

[50] [Reaction Scheme 3]

[51]

[52] [53] Another aspect of the present invention relates to a pharmaceutical composition for inhibiting cyclooxygenase-2 comprising the compound of the above formula (I) or pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable carrier. In particular, the pharmaceutical composition of the present invention can be used for treating or preventing inflammation, an allergic disorder, inflammatory dermatosis, an immunologic disorder, a neurodegenerative disorder, arthritis, pain or fever.

[54]

[55] The pharmaceutical composition according to the present invention can be administered orally, e.g., ingestion or inhalation; or parenterally, e.g., injection, deposition, implantation or suppositories. The injection can be, for example, intravenous, intradermal, subcutaneous, intramuscular or intraperitoneal. Depending on the route of administration, the pharmaceutical composition of the present invention may be formulated as tablets, capsules, granules, fine subtilae, powders, sublingual tablets, suppositories, ointments, injection solutions, emulsions, suspensions, syrups, aerosols, etc. The above various forms of the pharmaceutical composition of the present invention can be prepared in a manner well known in the art using a pharmaceutically acceptable carrier(s) which are usually used for each form. Examples of the pharmaceutically acceptable carriers include excipient, filler, expander, binder, disintegrator, lubricant, preservative, antioxidant, isotonic agent, buffer, coating agent, sweetening agent, dissolvent, base, dispersing agent, wetting agent, suspending agent, stabilizer, colorant, aromatic, etc.

[56] The pharmaceutical composition of the present invention contains 0.01 to 100 wt% of the compound according to the present invention or pharmaceutically acceptable salt thereof depending on the form thereof.

[57] The specific dosage of the present pharmaceutical composition can be varied with species of mammals including a human-being, route of administration, body weight, gender, age, severity of disease, judgment of doctor, etc. It is preferable that 0.01 to 50 mg of the active ingredient is administered per kg of body weight a day for oral use, while 0.01 to 10 mg of the active ingredient is administered per kg of body weight a day for parenteral use. The total daily dosage can be administered once or over several times depending on the severity of disease, judgment of doctor, etc.

[58]

Advantageous Effects

[59] The compounds of the present invention have excellent cyclooxygenase-2 inhibitory activity and low cytotoxicity. Therefore, the compounds of the present invention can be used for preparing pharmaceutical compositions for treating or preventing inflammation, an allergic disorder, inflammatory dermatosis, an immunologic disorder, a neurodegenerative disorder, arthritis, pain or fever. Best Mode for Carrying Out the Invention

[61] The present invention is further illustrated by the following examples, which are not to be construed to limit the scope of the invention.

[62]

[63] Example 1: Preparation of 3-benzori.31dioxol-5-yl-4-phenylfuran-2.5dione

(MCR-3004^s)

[64] Benzoylformic acid (1.67 g) and 3,4-(methylenedioxy)phenylacetic acid (2 g) were dissolved in acetic anhydride (20 ml) and refluxed for 3 hours. After the reaction was completed, the reaction mixture was allowed to cool to room temperature and treated with excessive water. Then, the reaction mixture was neutralized with a saturated NaHCO aqueous solution, extracted with ethyl acetate three times, and washed with a saline solution. The organic phase was dried with anhydrous magnesium sulfate (MgSO ) and distilled under reduced pressure. The resulting residue was recrystallized

4 with ethanol and methanol to give the target compound as a yellow solid (2.93 g, 90 %). [65] ¹H NMR(430 MHz, DMS0<i ) δ 7.50-7.44(5H, m, Ph), 7.06-7.00(2H, m, PhOCH

6 2

O), 6.89(1H, m, PhOCH O), 6.10(2H, s, PhOCH. O)

[66] ¹³C NMR(IOO MHz, DMS0<i ) δ 165.6, 149.9,^"l47.9, 138.4, 137.2, 13Q9, 129.8,

129.3, 128.2, 125.3, 121.4, 109.5, 109.3, 102.3.

[67]

[68] Example 2: Preparation of 3-benzo[1.31dioxol-5-yl-4-phenyl-l H -pyrrole -2.5dione

(MCR-3005)

[69] To a solution of the compound (0.68 g, 2.311 mmol) obtained in Example 1 dissolved in DMF (5 ml) were added HMDS (4.88 ml, 23.11 mmol) and methanol (0.47 ml, 11.555 mmol) in sequence, and the reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed, the reaction mixture was extracted with ethyl acetate three times. The organic phase was dried with anhydrous magnesium sulfate (MgSO ) and distilled under reduced pressure. The resulting

4 residue was subjected to silica gel column chromatography (n-hexane:EtOAc = 3:1) and recrystallized with ethanol and methanol to give the target compound as a yellow solid(0.61 g, 89 %). [70] ¹H NMR(430 MHz, DMSO<i ) δ 11.19(1H, s, -NH), 7.42-7.36(5H, m, Ph), 6.94(2H,

6 s, PhOCH O), 6.85(1H, s, PhOCH O), 6.05(2H, s, PhOCH. O)

2

[71] ¹¹³³CC NNMMRR((IIOOOO MMHHzz,, DDMM!S0<i ) δ 171.5, 148.3, 147.1, 1353, 129.5, 128.4, 124.4, 122.1, 109.3, 108.4, 101.4. [72]

[73] Example 3: Preparation of 3-G.4-dihydroxyphenylV4-phenyl-l H -pyrrole -2.5dione

(MCR-3006) [74] To a solution of the compound (0.06 g, Q 190 mmol) obtained in Example 2 dissolved in CH Cl was dropped BBr (1.14 ml, 1.142 mmol) at -78°C under nitrogen

2 2 3 atmosphere, and the reaction mixture was stirred for 16 hours. After the reaction was completed, the reaction mixture was slowly neutralized with a saturated NaHCO aqueous solution and extracted with ethyl acetate three times. The organic phase was dried with anhydrous magnesium sulfate (MgSO ) and distilled under reduced

4 pressure. The resulting residue was subjected to silica gel column chromatography (n-hexane: EtOAc = 3:1) to give the target compound as a yellow solid(0.01 g, 16 %). [75] ¹H NMR(430 MHz, DMS0<i ) δ 7.42-7.36(5H, m, Ph), 6.88(1H, d, J = 2.0 Hz, Ph

(OH) ), 6.73(1H, dd, J = 2.0 Hz, J = 8.4 Hz, Ph(OH) ), 6.68(1H, d, J= 8.4 Hz, Ph(OH)

2 2

[76] ¹³C NMR(IOO MHz, DMS0<i ) δ 172.0, 171.8, 147.3, 145.0, 136.5, 133.8, 129.5,

116.9, 115.5.

[77]

[78] Example 4: Preparation of 2-phenylacetamide

[79] To phenylacetic acid (4.25 g, 31.216 mmol) was dropped thionyl chloride (10 ml) at room temperature, and the reaction mixture was stirred for 6 hours. One drop of DMF was added and refluxed for 1 hour. After the reaction was completed, the solvent was evaporated under reduced pressure and the resulting residue was diluted with CH Cl . Then, NH OH (100 ml) was slowly added and stirred at room temperature for 12

4 hours. After the reaction was completed, the reaction mixture was extracted with CH Cl three times. The organic phase was dried with anhydrous magnesium sulfate (MgSO ) and distilled under reduced pressure. The resulting residue was recrystallized with CH Cl to give the target compound as a white solid (3.8 g, 90 %). [80] ¹H NMR(430 MHz, DMS0<i ) δ 7.47(1H, s, -CONH. ), 7.30-7.20(5H, m, Ph),

6 2

6.87(1H, s, -C0NH_ )

13 ²

[81] C NMR(IOO MHz, DMS0<i ) δ 172.2, 136.5, 129.0, 128.1, 126.2, 42.2.

[82]

[83] Example 5: Preparation of ethyl (4-thiomethoxyphenyl)oxoacetate

[84] To anhydrous CH Cl was dropped anhydrous AlCl (2.73 g, 21.97 mmol) and ethyl chlorooxoacetate (3.5 ml, 31.33 mmol) at -5 ⁰C. Afterwards, thioanisole (2.58 ml, 21.97 mmol) was dropped and stirred for 3 hours. After the reaction was completed, the reaction mixture was extracted with CH Cl three times, and the organic phase was

2 2 dried with anhydrous magnesium sulfate (MgSO ) and distilled under reduced

4 pressure. The resulting residue was subjected to silica gel column chromatography (n-hexane: EtOAc = 3:1) to give the target compound as a pale yellow oil(3.58 g, 73 %). [85] ¹H NMR (4)0 MHz, DMS0<i ) δ 7.91(2H, d, J= 7.8 Hz, Ph), 7.45(2H, d, J = 7.8

6

Hz, Ph), 4.45(2H, q, J = 7.2 Hz, OCH. CH ), 2.60(3H, s, PhSCH. ), 1.37(3H, t, J= 7.2 Hz, OCH CH_ )

2 3

[86] ¹³C NMR (100 MHz, DMS04 ) δ 187.3, 164.2, 136.0, 132.2, 130.2, 130.0, 62.8,

6

14.3.

[87]

[88] Example 6 Preparation of 3-(4-thiomethoxyphenylV4-phenyl-l H -pyrrole-2.5dione

(MCR-301 D

[89] To a solution of ethyl (4-thiomethoxyphenyl)oxoacetate(0.5 g, 2.229 mmol) obtained in Example 5 and 2-phenylacetamide(0.25 g, 1.850 mmol) obtained in Example 4 dissolved in anhydrous THF at 0 ⁰C was added NaH(0.2 g, 8.333 mmol), and the reaction mixture was stirred for 24 hours. Concentrated HC1(3 ml) was slowly added at O⁰C, and the reaction mixture was diluted with water and extracted with ethyl acetate three times. The organic phase was dried with anhydrous magnesium sulfate (MgSO )

4 and distilled under reduced pressure. The resulting residue was recrystallized with ethanol to give the target compound as a yellow solid (Q37 g, 68 %). [90] ¹H NMR(430 MHz, DMSO<i ) δ 11.2(1H, s, -NH), 7.42-7.37(5H, m, Ph), 7.35(2H,

6 d, J = 8.3 Hz, PhSCH ), 7.24(2H, d, J= 8.3 Hz, PhSCH ), 2.47(3H, s, PhSCH. )

3 3 3

[91] ¹³C NMR(IOO MHz, DMSO<i ) δ 172.1, 141.3, 136.5, 136.3, 130.5, 129.9, 129.4,

128.9, 125.6, 125.2, 14.5.

[92]

[93] Example 7: Preparation of 3-(44nethanesulfonylphenylV4-phenyl-l H - pyrrole-2.5dione (MCR-SOM)

[94] The compound (0.1 g, Q339 mmol) obtained in Example 6 was dissolved in CH Cl and cooled to 0 ⁰C. m-CPBA(meta-chloroperbenzoic acid) (Q 15 g, Q677 mmol) was dropped and stirred at 0 ⁰C for 2 hours. The reaction was quenched by adding 10 % Na S O and the reaction mixture was extracted with ethyl acetate and a saturated NaHCO

2 2 3 aqueous solution. The organic phase was washed with distilled water and a saline

3 solution, dried with anhydrous magnesium sulfate (MgSO ) and distilled under

4 reduced pressure. The resulting residue was subjected to silica gel column chro- matography (n-hexane: EtOAc = 1:1) to give the target compound as a pale yellow solid (Q09 g, 81 %). [95] ¹H NMR(4)0 MHz, DMSCki ) δ 11.4)(1H, s, -N H), 7.95(2H, d, J=8.43 Hz, Ph

SOCH ), 7.61(2H, d, J=8.43 Hz, PhSOCH ), 7.44-7.37(5H, m, Ph), 3.25(3H, s, -SCH.

3 3 3

[96] ¹³C NMR(IOO MHz, DMSO<i ) δ 171.7, 171.6, 141.7, 138.9, 134.5, 131.1, 13Q4,

130.2, 129.0, 128.7, 127.4, 43.7.

[97]

[98] Example 8: Preparation of 4-chlorosulfonylphenylacetic acid

[99] To chlorosulfonic acid(35 ml) was dropped phenylacetic acid(5 g, 36.724 mmol) at 0

⁰C, and the reaction mixture was stirred at 0 ⁰C for 1 hour and then at room temperature for 12 hours. After the reaction was completed, the reaction mixture was slowly cooled to 0 ⁰C and further stirred for 20 minutes. The resulting solid was filtered to give the target compound(7.8 g, 91 %).

[100] ¹H NMR (430 MHz, DMSO<i ) δ 7.55-7.47 (2H, m, Ph), 7.29-7.20 (2H, m, Ph), 3.58

6

(IH, s, -CH. COOH) 3.56 (IH, s, -CH. COOH)

13 ^{2 2}

[101] C NMR (100 MHz, DMS04 ) δ 172.5, 172.4, 147.6, 146.0, 135.6, 134.6, 129.7,

128.8, 127.7, 126.4, 125.4, 123.8.

[102]

[103] Example 9: Preparation of aminosulfonylphenylacetamide

[104] To a solution of the compound (3 g, 12.784 mmol) obtained in Example 8 dissolved in SOCl (30 ml) was dropped 6 drops of DMF. Afterwards, the reaction mixture was stirred at room temperature for 16 hours, and SOCl was evaporated under reduced pressure. The resulting residue was dissolved in acetonitrile and bubbled with NH gas at 0 ⁰C. Then, the reaction mixture was stirred at 0 ⁰C for 10 minutes and at room temperature for 3 hours in sequence. The resulting solid was filtered and the filtrate was distilled under reduced pressure. Then, distilled water was added and kept in a refrigerator. The resulting solid was filtered and the filtrate was saturated with a saline solution and extracted with ethyl acetate three times. The organic phase was dried with anhydrous magnesium sulfate(MgSO ) and distilled under reduced pressure to give a solid product. The solid product combined with the previously obtained solid was re- crystallized with acetone to give the target compound as a white solid(Q9 g, 33 %).

[105] ¹H NMR (430 MHz, DMS0<i ) δ 7.74 (2H, d, J = 7.4 Hz, Ph), 7.55 (IH, br, -CONH ), 7.42 (2H, d, J= 7.4, Ph), 7.30 (2H, br, -SO NH. ), 6.95 (IH, br, -CONH. ), 3.46

2 2 2 2

(2H, s, -CH. -) [106] ¹³C NMR (100 MHz, DMS04 ) δ 171.5, 142.1, 1415, 129.5, 125.5, 41.8.

[107]

[108] Example 10: Preparation of 3-(4-aminosulfonyrphenyl)-4-phenyl-l H - pyrrole-2.5dione (MCR-BOlS)

[109] To a solution of ethyl phenyloxoacetate(463 mg) and aminosulfonylpheny- lacetamide(355 mg) obtained in Example 9 dissolved in anhydrous THF(IO ml) at 0 ⁰C was added NaH(60%, 133 mg), and the reaction mixture was stirred for 24 hours. Concentrated HC1(5 ml) was slowly added at O⁰C, and the reaction mixture was diluted with water and extracted with ethyl acetate three times. The organic phase was dried with anhydrous magnesium sulfate(MgSO ) and distilled under reduced pressure. The

4 resulting residue was recrystallized with ethanol to give the target compound as a yellow solid(Q37 g, 68 %). [110] ¹H NMR(430 MHz, DMSCki ) δ 11.3(1H, s, -NH), 7.82(2H, d, J = 8.2 Hz, PhSO

6 2

NH ), 7.53(2H, d, J = 8.2 Hz, PhSO NH ), 7.44(1H, s, PhSO NH. ), 7.44-7.36(5H, m,

2 2 2 2 2

Ph)

13,

[111] ¹³C NMR(IOO MHz, DMSO<i ) δ 171.3, 172.0, 144.5, 137.9, 135.4, 132.2, 130.2,

129.8, 129.7, 129.6, 128.5, 128.3, 128.0, 125.9, 125.6, 62.Q

[112]

[113] Example 11 : Preparation of (4-thiomethoxyphenyl)oxoacetic acid [114] To the compound(4 g, 17.835 mmol) obtained in Example 5 was dropped 2N NaOH(8.9 ml) at room temperature, and the reaction mixture was stirred at room temperature for 3 hours. After the reaction was completed, the reaction mixture was acidified with concentrated HCl to pH 2 and extracted with CH Cl three times. The

2 2 organic phase was dried with anhydrous magnesium sulfate(MgSO ) and distilled

4 under reduced pressure. The resulting residue was recrystallized with CH Cl and hexane to give the target compound as a pale yellow solid(3.04 g, 87 %). [115] ¹H NMR(430 MHz, CDCl ) δ 8.23(2H, d, J= 8.6 Hz, Ph), 7.30(2H, d, J = 8.6 Hz, Ph), 2.54(3H, s, -SCH. )

3

[116] ¹³C NMR(IOO MHz, CDCl ) δ 183.6, 162.1, 150.2, 131.5, 128.0, 124.9, 14.5.

[117]

[118] Example 12: Preparation of 3-(4-thiomethoxyphenyl)-4-phenylfuran-2.5dione

(MCR-BOlO) [119] The compound obtained in Example 11 and phenylacetic acid were used according to the same procedure as Example 1 to give the target compound(69 %). [120] ¹H NMR (430 MHz, DMSO<i ) δ 7.50-7.45(5H, m, Ph), 7.34(2H, d, J = 8.5 Hz, Ph -

6 SCH ), 7.31(2H, d, J = 8.5 Hz, Ph-SCH ), 2.50(3H, s, -SCH. )

13 ^{3 3 2}

[121] C NMR (100 MHz, CDCl ) δ 165.1, 142.5, 137.7, 137.0, 13Q4, 129.7, 129.2,

128.7, 127.7, 125.1, 123.3, 13.8. [122] [123] Example 13: Preparation of 3-G.4.5-trimethoxyphenylV4-phenylfuran-2.5dione

(MCR-3007^s) [124] 3,4,5-trimethoxyphenylacetic acid was used according to the same procedure as

Example 1 to give the target compound(69 %). [125] ¹H NMR (430 MHz, DMSCki ) δ 7.50 (s, 5H, aromatic-C H_ ), 6.79 (s, 2H, C H. ),

3.72(s, 3H, -OCH ), 3.58(s, 3H, -OCH ) [126] ¹³C NMR (100 MHz, DMS04 ) δ 164.9, 152.6, 139.5, 137.8, 137.6, 130.4, 129.3,

6

128.7, 127.7, 122.4, 107.1, 60.1, 55.6.

[127]

[128] Example 14: Preparation of 3-(34nethoxyphenyl^s)-4-phenyl-l H -pyrrole -2.5dione (MCR-3002)

[129] To a solution of 3-(34nethoxypheynyl)-4-phenylfuran-2,5dione(1.022 g, 3.646 mmol) dissolved in methanol were added hexamethyldisilazane(5.885 g, 36.46 mmol) and DMF(5 ml), and the reaction mixture was stirred at room temperature for 24 hours. Then, the reaction mixture was neutralized with a saturated NaHCO aqueous solution and extracted with ethyl acetate. The organic phase was washed with a saturated saline solution, dried with anhydrous magnesium sulfate(MgSO ) and distilled under reduced pressure. The resulting solid was washed with petroleum ether and diethyl ether, and filtered to give the target compound as a yellow solid(0.468 g, 46 %).

[130] ¹H NMR (430 MHz, CDCl +DMSO-J ) δ 10.38 (s, IH, -N H), 7.46-7.24 (m, 5H,

3 6 aromatic-C H ),7.04~6.89 (m, 4H, aromatic-C H_ -OCH ), 3.67 (s, 3H, -OCH. )

6 5 6 4 3 3

[131] ¹³C NMR (100 MHz, CDCl +DMSO-J ) δ 171.5, 159.2, 136.9, 136.5, 129.8, 129.5,

3 6

128.6, 128.3, 122.1, 115.7, 114.7, 55.Q [132]

[133] Example 15: Preparation of 3.4-diphenyl-l H -pyrrole-2.5dione (MCR-SOOD [134] 3,4-diphenylfuran-2,5dione was used according to the same procedure as Example

14 to give the target compound(37 %). [135] ¹H NMR (430 MHz, CDCl +DMSO-J ) δ 10.88 (s, IH, -NH), 7.50-7.33 (m, ICH,

3 6 aromatic-C H )

6 5

[136] ¹³C NMR (100 MHz, CDCl +DMSO-J ) δ 170.9, 135.9, 129.0, 128.8, 128.0, 127.7.

3 6

[137] [138] Example 16 Preparation of 3-(3.4.5-trimethoxyphenyl^s)-4-phenyl-l H - pyrrole-2.5dione fMCR-SX)^ [139] 3-(3,4,5-trimethoxyphenyl)-4-phenylfuran-2,5dione obtained in Example 13 was used according to the same procedure as Example 14 to give the target compound(97

%). [143] ¹H NMR (430 MHz, DMSCM ) δ 11.27 (br, IH, -NH), 7.42 (s, 5H, aromatic-C H_

6 6 5

), 6.72 (q, 2H, aromatic-C H_ ), 3.69-3.67 (q, 3H, -OCH. ), 3.56 (s, 6H, -OCH. ) [141] ¹³C NMR (100 MHz, DMSO-d ) δ 172.1, 171.9, 152.9, 139.1, 136.7, 136.5, 13Ql^",

129.8, 129.4, 128.8, 124.3, 107.7, 6Q6, 56.1. [142] [143] Example 17: Preparation of 3-(3-hvdroxyphenyl^s)-4-phenyl-l H -pyrrole-2.5dione

(MCR-3003) [144] To a solution of 3-(34nethoxyphenyl)-4-phenyl-l H-pyrrole-2,5dione(0.2 g, Q716 mmol) obtained in Example 14 dissolved in CH Cl (24 ml) was slowly added BBr

2 2 3

(0.179 g, 0716 mmol) at -78 °C, and the reaction mixture was left to room temperature and stirred for 12 hours. After the reaction was completed, the reaction mixture was neutralized with a saturated NaHCO aqueous solution and extracted with ethyl acetate. The organic phase was washed with a saturated saline solution, dried with anhydrous magnesium sulfate(MgSO ) and distilled under reduced pressure to give the

4 target compound as a yellow solid(100 mg, 50%). [145] ¹H NMR (430 MHz, CDCl ) δ 10.8 (s, IH, -NH), 7.46-7.43 (m, 5H, aromatic-C H

3 6

[ 171.6, 129.8, 129.3 128.9 128.2 12Q5 116.5

6Q 1.

[147]

[148] Example 18: Preparation of 3-(3A5-trihvdroxyphenyl)-4-phenyl-l H - pyrrole-2.5dione (MCR- 3009^s) [149] 3-(3,4,5-trimethoxyphenyl)-4-phenyl-l H-pyrrole-2,5dione obtained in Example 16 was used according to the same procedure as Example 17 to give the target compound(88 %). [150] ¹H NMR (430 MHz, DMSO<i ) δ 11.02 (IH, s, -NH), 9.00 (2H, s, OH), 8.61 (IH, s,

6

OH), 7.39 (5H, s, Ph), 6.42 (2H, s, Ph(OH) ) [151] ¹³C NMR (100 MHz, DMS04 ) δ 172.4, 172.3, 146.2, 137.2, 135.9, 134.2, 13Ql,

129.9, 129.5, 128.8, 119.0, 109.5. [152] [153] Example 19: Preparation of

3 - (4-acetoxyphenyl V 4- (44nethanesulf onyrphenyl^s)furan-2.5dione (MCR- 3018^s)

[154] (4-Thiomethoxyphenyl)oxoacetic acid obtained in Example 11 and

(4-acetoxyphenyl)acetic acid were used according to the same procedure as Example 1 to provide 3-(4-acetoxyphenyl)-4-(4-thiomethoxyphenyl)furan-2,5dione, which was used according to the same procedure as Example 7 to give the target compound(26 %).

[155] ¹H-NMR (430 MHz, DMSCki ) δ 8.04(2H, d, J = 8.8 Hz, aromatic-C H ), 7.69(2H,

6 6 2 d, J = 8.8 Hz, aromatic-C H ), 7.50(2H, d, J = 8.6 Hz, aromatic-C H ), 7.27 (2H, d, J

6 2 6 2

= 8.6 Hz, aromatic-C H ), 3.28(3H, s, PhSO CH ), 2.28(3H, s, PhOCOCH ).

6 2 2 3 3

[156]

[157] Example 20: Preparation of 3-(4-hydroxyphenylV4-(4-thiomethoxyphenyl^s) -I H - pyrrole-2.5dione (MCR-SO^

[158] (4-Thiomethoxyphenyl)oxoacetic acid obtained in Example 11 and

(4-acetoxyphenyl)acetic acid were used according to the same procedure as Example 1 to provide 3-(4-acetoxyphenyl)-4-(4-thiomethoxyphenyl)furan-2,5dione, which was used according to the same procedure as Example 2 to give the target compound(30 %).

[159] ¹H-NMR (430 MHz, DMS04 ) δ 11.08(1H, s, OH), 7.33(2H, d, J = 6.8Hz,

6 aromatic-C H ), 7.32-7.24(4H, m, aromatic-C H ), 6.68-6.74(2H, m, aromatic-C H

6 2 6 4 6 2

), 2.48(3H,s, C H S). [160] [161] Example 21: Preparation of

3-(4-acetaminophenyl)-4-(4-thiomethoxyphenyl)furan-2.5dione (MCR- 3026) [162] (4-Thiomethoxyphenyl)oxoacetic acid obtained in Example 11 and

(4-acetaminophenyl)acetic acid were used according to the same procedure as

Example 1 to give the target compound(59 %). [163] ¹H-NMR (430 MHz, DMS04 ) δ 1Q21(1H, s, NH), 7.65(2H, d, J = 8.8 Hz,

6 aromatic-C H ), 7.42(4H, t, J = 8.0 Hz, aromatic-C H ), 7.33(2H, d, J = 8.8 Hz,

6 2 6 4 aromatic-C H ), 2.50(3H, s, SCH ), 2.07(3H, s, PhNHCOCH ).

6 2 3 3

[164]

[165] The chemical structures and spectroscopic data of the representative compounds of the present invention which were prepared in the above Examples are summarized in

Table 1. [166] Table 1

[169] [170] Biological Activity Test [171] For the biological activities of the compounds of the present invention, the concentration to inhibit the production of prostaglandin E2(PGE 2 ) by 50% in Raw264.7 murine macrophage cell line treated with lipopolysacchande(LPS) (IC ) was analyzed, and for the compounds showing less than 10 μM of IC in the above test, the concentration to inhibit the activity of COX-2 enzyme which is associated with the production of PGE by 50% (IC ) was tested. Also, for the cytotoxicities of the

2 50 compounds according to the present invention, cell viability (IC ) was measured according to a 3-(4,5dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide (MTT) assay.

[172]

[173] Experiment 1: Inhibition of LPS -induced PGE 2 production

[174] Raw264.7 cells(murine macrophage cell line), which were obtained from Korean Cell Line Bank(KCLB), were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS), penicillin(100 units/mL) and streptomycin sulfate(10Q7g/mL) at 37 ⁰C in a humidified atmosphere with 5% CO . 1 mL of Raw264.7 cells in DMEM(5xl0 cells/mL) was seeded into each well of 24-well plates, left overnight and replaced with a fresh medium. Various concentrations of compounds were added and then incubated for 1 hour. Lipopolysaccharide(LPS) was added at lμg/mL and incubated for 24 hours(or appropriate time). The obtained supernatant was diluted five fold. 150μL of analysis buffer was added to non specific binding(NSB) wells and lOOμL of analysis buffer was added to zero standard(BO) wells. lOOμL of standard samples were added to the other wells. 50μL of PGE conjugates were added (except for NSB). Then, 50μL of PGE antibody solution was added and shaken for 2 hours. Each well was subject to suction and washed with washing buffer five times. 200μL of para-nitrophenyl phosphate (pNPP) substrate was added to each well and left at room temperature for 1 hour in bench, and then 50μL of stop solution was added. The absorbance was measured at 4)5 nm. The production of PGE 2 was evaluated using the measured absorbance and standard curve, and the con- centration to inhibit by 50% (IC ) compared to the group treated with only LPS was analyzed. The results are summarized in Table 2, where the values were obtained from three independent experiments.

[175]

[176] Experiment 2: Inhibition of COX-2 activity

[177] To a background tube were added 970μL of reaction buffer and lOμL of heme, and then lOμL of COX-2 protein inactivated by boiling for 3 minutes. To a COX-2 100% initial activity tube were added 970μL of reaction buffer and lOμL of heme, and then lOμL of COX-2 protein. To a COX-2 inhibition tube were added 950μL of reaction buffer and lOμL of heme, and then lOμL of COX-2 protein and 20μL of standard sample. All tubes were left at 37 ⁰C for 10 minutes. lOμL of arachidonic acid was added and vortexed, and then left at 37 ⁰C for 2 minutes. 50μL of IM HCl and lOOμL of saturated tin chloride solution were added in sequence and vortexed, and then left at room temperature for 5 minutes. lOOμL of Enzyme ImmunoAssay(EIA) buffer was added to non specific binding wells and 50μL of EIA buffer was added to zero standard(BO) wells. Background sample, COX-2 100% initial activity sample diluted 4)00: 1 and COX-2 inhibition sample were added in 50μL aliquots to the other wells. Prostaglandin screening tracer was added in 50μL aliquots except for total activity(TA) and Bik-like killer(Blk). PG screening antiserum was added in 50μL aliquots except for TA, NSB and BIk, and left at room temperature for 18 hours. Each well was subject to suction and washed with washing buffer 5 times. Ellman's reagent was added in 200μL aliquots and tracer was added in 5μL aliquots to only TA wells. The plates were shaken for 60-90 minutes with shielding the light and the absorbance was measured at 4)5 nm. The inhibition of COX-2 activity was evaluated using the measured absorbance and standard curve, and the concentration to inhibit by 50% (IC

50

) compared to the group of COX-2 100% initial activity samples was analyzed. The results are summarized in Table 2, where the values were obtained from three independent experiments.

[178]

[179] Experiment 3: Cytotoxicities

[180] Raw264.7 cells(murine macrophage cell line) were cultured in Dulbecco's modified Eagle's medium(DMEM) containing 10% fetal bovine serum(FBS), penicillin(100 units/mL) and streptomycin sulfate(100 //g/mL) at 37 ⁰C in a humidified atmosphere with 5% CO . The cells were collected by centrifugation and scraper, and added in 1x10 cells/well to 96- well plates having lOOμL of Roswell Park Memorial Institute(RPM) 164) medium containing 10% FBS.

3-beta,4-beta-epoxy-8a-isobutyryloxyguaia- 1 ( 10), 11 ,( 13)-diene- 12.6aolide was dissolved in dimethylsulfoxide(DMSO) and the concentration of DMSO did not exceed Ql% for all tests. After overnight, samples and LPS(I //g/mL) were added and the plates were incubated for 24 hours. The cells were washed once and 50 μL of medium without FBS containing 5 mg/mL of

3-(4,5dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide(MTT) was added. Then, the plates were incubated at 37 ⁰C for 4 hours. Afterwards, the medium was removed and formazan blue formed in the cells was dissolved in 100 μL of DMSO. The absorbance was measured at 54) nm to analyze the cytotoxicities in IC values. IC

50 50 means the concentration to exhibit 50% reduction in the number of cells compared to the group which was not treated with compounds. The results are summarized in Table 2, where the values were obtained from three independent experiments.

[181] [182] Table 2 [183]

PGE2 COX-2 PGE2 COX-2

Cell Cell

Compound Product ion Activity Compound Production Activity Viability Viabi lity Code Inhibit ion Inhibition Code Inhibition Inhibition IC₅₀ (liM) IC₅₀ (iiM) IC₅₀ ⁽UM⁾ IC₅₀(UM) IC₅₀(UM) IC₅₀ (uM)

MCR3001 269.90 27.94 ND MCR3016 333.94 30.46 ND

MCR3002 259.63 9.95 ND MCR3017 261.90 68.75 ND

MCR3003 158.01 13.83 ND MCR3018 261.82 4.41 16.63

MCR3004 328.68 7.13 18.52 MCR3019 124.74 0.84 21.01

MCR3005 195.36 7.96 ND MCR3020 266.08 12.6 ND

MCR3006 206.59 2.69 16.04 MCR3021 283.66 23.75 ND

MCR3007 312.56 37.1 ND MCR3022 271.24 - ND

MCR3008 150.55 25.08 ND MCR3023 121.22 25.5 ND

MCR3009 261.45 8.19 17.51 MCR3024 242.04 45.2 ND

MCR3010 194.77 3.58 18.45 MCR3025 260.76 10.76 ND

MCR3011 151.88 4.73 16.34 MCR3026 249.75 5.71 20.11

MCR3012 303.22 21.52 ND MCR3027 276.61 78.48 ND

MCR3013 265.89 15.42 ND MCR3028 257.40 71.29 ND

MCR3014 268.74 2.71 17.5 MCR3029 305.41 131.53 ND

MCR3015 158.18 0.61 15.62

Claims

[1] A compound of formula (I) or pharmaceutically acceptable salt thereof:

(I) wherein,

R , R and R are each independently hydrogen, hydroxy, C -C lower alkoxy, R

1 2 3 1 5 5

C(=O)O, R S, R S(=O), R S(=O) , NH S(=O) ,R C(=O)NH, or (R C(=O)) N; or

5 5 5 2 2 2 5 5 2

R and R taken together with the carbon atoms to which they are attached form a

2 3 heterocycle;

R is hydrogen, R S, R S(=O) or R S(=O) ;

4 5 5 5 2

R is C -C lower alkyl; and

5 1 5

X is O or NH.

[2] The compound according to claim 1 or pharmaceutically acceptable salt thereof, wherein R , R and R are each independently hydrogen, hydroxy, C -C lower alkoxy, R C(=O)O, R S, R S(=O) , NH S(=O) , or R C(=O)NH.

5 5 5 2 2 2 5

[3] The compound according to claim 2 or pharmaceutically acceptable salt thereof, wherein R is methyl.

[4] The compound according to claim 1 or pharmaceutically acceptable salt thereof, wherein R is hydrogen; R and R taken together with the carbon atoms to which they are attached form dioxolane or dioxane. [5] The compound according to claim 1 or pharmaceutically acceptable salt thereof, wherein R , R and R are each independently hydrogen, hydroxy, or C -C lower

1 2 3 1 5 alkoxy; or R and R taken together with the carbon atoms to which they are

2 3 attached form dioxolane or dioxane; and R is hydrogen.

4

[6] The compound according to claim 1 or pharmaceutically acceptable salt thereof, wherein R and R are hydrogen; R is hydroxy, C -C lower alkoxy, R C(=O)O,

1 3 2 1 5 5

R S, R S(=O), R S(=O) , NH S(=O) ,R C(=O)NH, or (R C(=O)) N; and

5 5 5 2 2 2 5 5 2

R is hydrogen.

4

[7] The compound according to claim 6 or pharmaceutically acceptable salt thereof, wherein R is methyl.

[8] The compound according to claim 1 or pharmaceutically acceptable salt thereof, wherein R and R are hydrogen; R is hydrogen, hydroxy, C -C lower alkoxy, R C(=O)O, R S, R S(=O), R S(=O) , NH S(=O) ,R C(=O)NH, or (R C(=O)) N;

5 5 5 5 2 2 2 5 5 2 and

R is R S, R S(=O) or R S(=O) .

4 5 5 5 2

[9] The compound according to claim 8 or pharmaceutically acceptable salt thereof, wherein R is methyl. [10] The compound according to claim 1 or pharmaceutically acceptable salt thereof, wherein the compound is selected from the following group:

3-(34nethoxyphenyl)-4-phenyl- 1 H-pyrrole-2,5dione;

3 -benzo [1,3] dioxol-5-yl-4-phenylfuran-2,5dione ;

3 -benzo [1,3] dioxol-5-yl-4-phenyl- 1 H-pyrrole-2,5dione ;

3-(3,4-dihydroxyphenyl)-4-phenyl- 1 H-pyrrole-2,5dione;

3-(3,4,5-trihydroxyphenyl)-4-phenyl- 1 H-pyrrole-2,5dione;

3 - (4-thiomethoxyphenyl) -4-phenyrfuran-2,5dione ;

3 - (4-thiomethoxyphenyl) -4-phenyl- 1 H-pyrrole-2,5dione ;

3-(44nethanesulfonylphenyl)-4-phenyl- 1 H-pyrrole-2,5dione;

3 -(4-aminosulfonylphenyl) -4-phenyl- 1 H-pyrrole-2,5dione;

3-(4-acetoxyphenyl)-4-(4-methanesulfonylphenyl)furan-2,5dione;

3 - (4-hydroxyphenyl) -4- (4-thiomethoxyphenyl) - 1 H-pyrrole-2,5dione ; and

3-(4-acetaminophenyl)-4-(4-thiomethoxyphenyl)furan-2,5dione. [11] A pharmaceutical composition for inhibiting cyclooxygenase-2(COX-2) comprising the compound of formula (I) according to any one of claims 1 to 10 or pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable carrier. [12] The pharmaceutical composition according to claim 11 for treating or preventing inflammation, an allergic disorder, inflammatory dermatosis, an immunologic disorder, a neurodegenerative disorder, arthritis, pain or fever.