KR101056878B1 - Novel Jasmonate Derivatives and Their Pharmaceutical Uses - Google Patents

Abstract

The present invention relates to a novel jasmonate derivative represented by the following formula (1) and its use, which is responsible for the production of inflammatory mediators such as NO, IL-6 and TNF-α in LPS-activated RAW 264.7 cells. By inhibiting, it can be very usefully used as a therapeutic agent for inflammatory diseases and a therapeutic agent for immune diseases.

[Formula 1]

Jasmonate, anti-inflammatory, immunosuppressant, NO, IL-6, TNF-α

Description

New jasmonate derivatives and pharmaceutical use thereof

The present invention inhibits the production of inflammatory mediators such as NO, IL-6 and TNF-α in LPS-activated RAW 264.7 cells, and thus is a novel jasmonate derivative that can be usefully used as a therapeutic agent for inflammatory diseases and immune diseases. And medicinal uses thereof.

The regulation of inflammatory responses is closely related to the pathogenesis of inflammation-related diseases, which are associated with the sequential activation of various signaling such as cyclooxygenase, NO synthase, cytokines and the like.

Excessive production of inflammatory mediators, including NO, IL-6 and interleukins such as IL-1β and TNF-α, mediates inflammatory diseases such as rheumatoid arthritis, inflammatory bowel disease, osteoporosis, psoriasis, endotoxins, and toxic shock syndrome.

In addition, such inflammatory mediators include autoimmune diseases such as organ transplantation, rheumatoid arthritis, myasthenia gravis, multiple sclerosis, Sjogren's syndrome, insulin dependent diabetes mellitus, systemic lupus erythematosus and the like; Or mediate immune-related diseases associated with allergies such as atopy and allergic rhinitis.

Inflammatory responses occurring in the human body can be classified into acute inflammation and chronic inflammation. An acute inflammatory response is a rapidly occurring reaction that protects the body from foreign antigens.

The onset of an acute inflammatory process is the secretion and induction of inflammatory cytokines such as interleukin-1 beta, interleukin-6, and tumor necorsis factor-alpha (TNF-α). It is accompanied by an increase in inflammatory mediator genes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2).

Macrophages activated by these secreted inflammatory regulators effectively remove the influx of antigen, and once all antigens are removed, further progression stops.

Therefore, it is very unlikely that an acute inflammatory response will develop into human disease, but in some chronic inflammatory diseases, such as pertussis, the expression of IL-23 increases, leading to chronic inflammation ( Infect. Immun. , 73 : 1590-1597, 2005).

Unlike acute, the chronic inflammatory response is an inflammatory response that proceeds for a long time by internal factors rather than external antigens. It is not an inflammatory response caused by macrophages but an inflammatory response involving macrophages, neutrophils and T lymphocytes. Increased expression of 23, resulting in the continuous expression of inflammatory cytokines such as TNF-α ( Trends in Immunol , 27 : 17-23, 2006).

Treatment of these chronic inflammatory diseases has traditionally been an inhibitor of inflammation by steroids such as prednisolone, nonsteroidal anti-inflammatory drugs (NASAIDs) such as naproxen, and calcium and calmodulin dependent protein phosphorylation such as cyclosporine or FK506. Immunosuppressants that bind to the enzyme calcineurin and inhibit its action have been most used.

However, these steroids and immunosuppressive agents are accompanied by side effects such as renal toxicity, infection, lymphoma, diabetes, tremor, headache, diarrhea, high blood pressure, nausea, renal dysfunction.

Therefore, there is an urgent need to develop an effective drug that suppresses the overexpression of inflammatory mediators while reducing side effects of currently used inflammatory diseases and immunosuppressive agents.

It is an object of the present invention to inhibit the production of inflammatory mediators such as NO, IL-6 and TNF-α in LPS-activated RAW 264.7 cells, which can be usefully used as a therapeutic agent for inflammatory and immune diseases. Is in providing.

In addition, another object of the present invention to provide a pharmaceutical composition for the prevention and treatment of inflammatory diseases and immune diseases containing a novel jasmonate derivative as an active ingredient.

In order to achieve the above object, the present invention provides a jasmonate derivative represented by the following formula (1):

[Formula 1]

R ₁ is any one selected from OCH ₃ , OH, NHCH (CH ₃ ) ₂ or NHC ₂ H ₄ OH,

R ₂ is any one selected from hydrogen, CN, COOCH ₃ or halogen,

The bond between C-4 and C-5 and C-9 and C-10 is a single bond or a double bond with a dashed line.

The derivatives are methyl 4,5-didihydrozasmonate (compound 2), methyl 5-cyano-4,5-didihydrozasmonate (compound 3), methyl 5-methoxycarbonyl-4,5- Didihydrozasmonate (compound 4), 4,5-didihydrozasmonate (compound 5), methyl 5-chloro-4,5-didihydrozasmonate (compound 7), 5-chloro-4,5 Didihydrozasmonic acid (compound 8), methyl 5-bromo-4,5-didihydrozasmonate (compound 9), methyl 5-iodo-4,5-didihydrozasmonate (compound 10), Methyl 4,5-didihydro-dihydrozasmonate (Compound 11), 4,5-didihydro-dihydrozasmonic acid (Compound 12), methyl 5-chloro-4,5-didihydro-dihydrozasmo Nate (Compound 13), 5-Chloro-4,5-didihydro-dihydrozasmonic acid (Compound 14), Methyl 5-bromo-4,5-didihydro-dihydrozasmonate (Compound 15), methyl 5- Iodo-4,5-didihydro-dihydrozasmonate (Compound 16), N-isopropyl-4,5-didihydrozasmonylamide (Compound 17), N-hydroxyethyl-4,5- Didihydrozasmonilamide (Compound 18), N-isopropyl-4,5-didihydro-dihydrozasmonilamide (Compound 19) and N-hydroxyethyl-4,5-didihydro-dihydroza It is preferably any one selected from the group consisting of smonylamide (compound 20).

Further, the derivative is more preferably R ₁ is OCH ₃ , R ₂ is halogen, and more specifically methyl-5-chloro-4,5-didihydrozasmonate; Methyl-5-bromo-4,5-didihydrozasmonate; Methyl-5-chloro-4,5-didihydro-dihydrozasmonate; And methyl-5-bromo-4,5-didihydro-dihydrozasmonate.

The present invention also provides a pharmaceutical composition for preventing and treating inflammatory diseases, which contains the jasmonate derivative represented by Chemical Formula 1 as an active ingredient.

The present invention also provides a pharmaceutical composition for the prevention and treatment of immune diseases, which contains the jasmonate derivative represented by the formula (1) as an active ingredient.

The present invention also provides a health food for improving an inflammatory disease or an immune disease containing a jasmonate derivative represented by the formula (1) as an active ingredient.

Hereinafter, the present invention will be described in more detail.

The present inventors confirmed that the newly synthesized jasmonate derivatives exhibited the activity of inhibiting the production of inflammatory mediators such as NO, IL-6 and TNF-α in LPS-activated RAW 264.7 cells, thereby completing the present invention. It became.

As shown in Figure 1, Compound 2 shows a significantly improved inflammatory mediator inhibitory activity than Compound 1, which is a known methyl jasmonate, it was confirmed the importance of the enone group in the life of the jasmonate derivative.

In addition, as shown in Figures 2 and 3, due to halogen substitution at the α-position of the enone group shows a more excellent inflammatory mediator inhibitory activity, as shown in Figure 4 is more effective than the natural anti-inflammatory prostaglandin Excellent.

Jasmonate derivative of the present invention is preferably contained in 5 to 10 parts by weight based on 100 parts by weight of the pharmaceutical composition for preventing and treating inflammatory diseases or immune diseases. If the derivative is contained in a small amount out of the above range may cause a decrease in the efficacy of the drug, while containing a large amount may cause side effects such as abdominal pain, diarrhea, nausea, vomiting.

In addition, the pharmaceutical composition according to the present invention may further include a suitable carrier, excipient or diluent commonly used in the preparation of the pharmaceutical composition.

Carriers, excipients or diluents usable in the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method .

When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid form preparations include at least one excipient such as starch, calcium carbonate, sucrose ( Prepare by mixing sucrose or lactose, gelatin, etc.

In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The amount of the jasmonate derivative used in the present invention may vary depending on the age, sex, and weight of the patient, but may be administered once to several times daily in an amount of 2 to 10 mg / kg.

In addition, the dose of the jasmonate derivative may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, humans by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The jasmonate derivative of the present invention was confirmed as a safe compound having an ID ₅₀ of 20 mg / kg or more as a result of acute toxicity test by intraperitoneal administration in mice.

The present invention also provides a health food for improving an inflammatory disease or an immune disease containing a jasmonate derivative represented by the formula (1) as an active ingredient.

The health food is used with other food or food additives in addition to the jasmonate derivative of the present invention, and may be suitably used according to a conventional method. The mixed amount of the active ingredient may be appropriately determined depending on the purpose of use thereof, for example, prophylactic, health or therapeutic treatment.

The effective dose of the compound contained in the health food may be used in accordance with the effective dose of the therapeutic agent, but may be less than the above range in the case of long-term intake for health and hygiene purposes or health control purposes It is evident that the component can be used in an amount above the range because there is no problem in terms of safety.

There is no particular limitation on the kind of the health food, for example, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, drinks, tea, Drinks, alcoholic drinks, vitamin complexes, etc. are mentioned.

Jasmonate derivatives according to the present invention inhibit the production of inflammatory mediators such as NO, IL-6 and TNF-α in LPS-activated RAW 264.7 cells, thereby rheumatoid arthritis, inflammatory bowel disease, osteoporosis, psoriasis, endotoxinemia It can be used as a therapeutic agent for inflammatory diseases mediated by inflammatory mediators such as toxic shock syndrome, and also for organ transplantation, rheumatoid arthritis, myasthenia gravis, multiple sclerosis, Sjogren's syndrome, insulin dependent diabetes mellitus, systemic Autoimmune diseases such as lupus erythematosus and the like; Or it can be usefully used as a therapeutic agent for immune-related diseases associated with allergies such as atopic dermatitis and allergic rhinitis.

Preferred examples are provided below to aid in the understanding of the present invention. However, these examples are only provided to more easily understand the present invention, the present invention is not limited to the following examples.

Example 1 Methyl 4,5-didihydrojasmonate (Methyl 4,5-didehydrojasmonate; 2)

To a stirred solution of methyl jasmonate (50 mg; 1) dissolved in DMSO (5 mL) was added IBX ( o- iodoxybenzoic acid, 2.0 equiv) and trifluoroacetic acid (0.3 equiv). The resulting mixture was stirred for 48 h at 80 ° C., then the cooled reaction mixture was diluted with saturated NaHCO ₃ solution and extracted with EtOAc (3 times).

The combined organic phases were washed with water and brine. After drying with MgSO ₄ , the organic layer was concentrated. The resulting residue was purified by reverse phase HPLC (YMC ODS-H80) eluted with 65% water soluble CH ₃ CN to give Compound 2 (15 mg, 30%).

¹ H and ¹³ C spectra for the compounds prepared in the Examples, including the above compounds, were recorded on a Varian Inova 400 spectrometer, and chemical shifts were determined for the relevant residual solvent or deuterium solvent peaks (δ _H 7.24 and δ _C 77.0 for CDCl ₃ ). FABMS data was obtained from a JEOLJMS SX-102A spectrometer, and HRFABMS data was obtained from a JEOLJMS SX-101A spectrometer. In addition, HPLC was performed on a YMC ODS-H80 column (250 × 10 mm, 4 μm, 80 μs) and a C18-5E Shodex packed column (250 × 10 mm, 5 μm, 100 μs) using a Shodex RI-71 detector.

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.61 (1H, dd, J = 6.0, 2.8 Hz, H-4), 6.17 (1H, dd, J = 6.0, 2.4 Hz, H-5), 5.46 ( 1H, m, H-10), 5.24 (1H, m, H-9), 3.70 (3H, s, OCH ₃ ), 2.99 (1H, m, H-3), 2.57 (1H, dd, J = 15.6 , 6.8 Hz, H-2b), 2.532.49 (1H, m, H-8b), 2.45 (1H, dd, J = 15.6, 8.4 Hz, H-2a), 2.29 (1H, m, H-8a) , 2.06 (3H, m, H-7 and H-11), 0.94 (3H, t, J = 8.0 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 210.2 (C-6), 171.8 (C-1), 165.4 (C-4), 134.5 (C-10), 133.7 (C-5), 124.4 (C -9), 51.8 (OCH ₃ ), 50.9 (C-7), 43.2 (C-3), 38.1 (C-2), 27.6 (C-8), 20.5 (C-11), 14.1 (C-12 ) FABMS m / z 223 [M + H] ⁺ HRFABMS m / z 223.1345 [M + H] ⁺ .

Example 2 Methyl 5-hydroxymethylene-jasmonate; 1a)

Several drops of methyl jasmonate (50 mg; 1) were added to a stirred suspension of NaH (3.0 equiv) and a catalytic amount of MeOH dissolved in anhydrous toluene (50 mL) at 0 ° C. After stirring at 0 ° C. for 10 minutes, several drops of ethyl formate (7.0 equiv) were added and the reaction mixture was stirred overnight at room temperature. Then, the aqueous solution extracted with cold water was extracted once more with ether and the unreacted starting material was removed. This aqueous solution was acidified with 10% HCl and the orange oil layer was removed by extraction with ether. This ether solution was washed with dilute NaHCO ₃ to remove excess acid and dried over MgSO ₄ .

After evaporation of the ether solution, the obtained residue (48 mg, 86%) was used for the next reaction without purification.

Yellow oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.17 (1H, s), 5.47 (1H, m), 5.30 (1H, m), 3.68 (3H, s), 2.75 (1H, m), 2.62 (1H , m), 2.442.14 (6H, m), 2.03 (2H, m), 0.95 (3H, t, J = 7.6 Hz).

<Example 3> O, N -Bis- (trifluoroacetyl) -hydroxyamine [ O, N -Bis- (trifluoroacetyl) -hydroxylamine]

O, N -bis- (trifluoroacetyl) -hydroxyamine was prepared by refluxing 3.2 moles of anhydrous trifluoroacetic acid with 1 mole of hydrochloric acid hydroxyamine for 1.5 hours. After trifluoroacetic acid and trifluoroacetyl chloride were removed under vacuum, the residue was recrystallized from CH ₂ Cl ₂ . The obtained product was O, N -bis- (trifluoroacetyl) -hydroxyamine with moisture absorption and volatile beading (80%).

Example 4 Methyl 5-cyano-jasmonate (1b)

To the previously prepared O, N -bis- (trifluoroacetyl) -hydroxyamine (3.0 equiv) was added a solution of compound 1a (20 mg) and pyridine (6.0 equiv) dissolved in toluene, and the resulting mixture was stirred for 1 hour. Heated under reflux. After cooling, the organics were diluted with EtOAc, washed with H ₂ O, washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure.

The crude product was purified by reverse phase HPLC (YMC ODS-H80) eluted with 50% water soluble MeOH to give compound 1b (6.0 mg, 30%).

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 5.51 (1H, m), 5.21 (1H, m), 3.70 (3H, s), 3.40 (1H, dd, J = 8.4, 5.6 Hz, minor isomer), 3.10 (1H, dd, J = 12.8, 8.4 Hz, major isomer), 2.73 (1H, m), 2.562.29 (6H, m), 2.13 (1H, m), 2.03 (2H, m), 0.95 (3H , t, J = 7.2 Hz, minor isomer), 0.93 (3H, t, J = 7.2 Hz, major isomer).

Example 5 Methyl 5-cyano-4,5-didehydrojasmonate (3)

A mixture of compound 1b (5 mg) and 2,3-dichloro-5,6-dicyanobenzoquinone (DDQ, 2.0 equiv) dissolved in anhydrous toluene was heated under reflux for 10 minutes. Insoluble material was removed by filtration through a cotton swab clogged pipette. The filtrate was concentrated in vacuo to give a residue, which was purified by reverse phase HPLC (YMC ODS-H80) eluted with 70% aqueous MeOH to give compound 3 (1.4 mg, 28%).

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 8.23 (1H, d, J = 2.4 Hz, H-4), 5.52 (1H, m, H-10), 5.19 (1H, m, H-9), 3.71 (3H, s, OCH ₃ ), 3.13 (1H, m, H-3), 2.68 (1H, dd, J = 16.0, 6.0 Hz, H-2b), 2.54 (2H, m, H-2a and H -8b), 2.38 (1H, m, H-8a), 2.26 (1H, m, H-7), 2.02 (2H, m, H-11), 0.95 (3H, t, J = 7.6 Hz, H- 12) FABMS m / z 248 [M + H] ⁺ HRFABMS m / z 248.1314 [M + H] ⁺ .

Example 6 Methyl 5-methoxycarbonyl-jasmonate; 1c)

Methyl jasmonate (20 mg; 1) was added to a stirred suspension of NaH (2.5 equiv) and a catalytic amount of MeOH dissolved in toluene at room temperature. After stirring for 15 minutes, dimethyl carbonate (0.5 mL) was added and the reaction mixture was heated under reflux for 3 hours. The reaction mixture was then stirred overnight, acidified with 10% HCl and extracted with EtOAc (3 times). The combined extracts were washed with water and brine, dried over MgSO ₄ and filtered. The solvent was removed under reduced pressure and the crude product was purified by reverse phase HPLC (YMC ODS-H80) eluted with 70 % aqueous CH ₃ CN to give compound 1c (12.5 mg, 50%).

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 5.45 (1H, m), 5.21 (1H, m), 3.74 (3H, s), 3.70 (3H, s), 3.34 (1H, dd, J = 9.6, 4.0 Hz, minor isomer), 3.13 (1H, dd, J = 12.4, 8.4 Hz, major isomer), 2.70 (1H, m), 2.592.18 (6H, m), 2.132.00 (3H, m), 0.94 (3H, t, J = 7.6 Hz).

Example 7 Methyl 5-methoxycarbonyl-4,5-didihydrojasmonate (Methyl 5-methoxycarbonyl-4,5-didehydrojasmonate; 4)

Compound 1c (5 mg) and 2,3-dichloro-5,6-dicyanobenzoquinone (DDQ, 2.0 equiv) dissolved in anhydrous toluene were heated under reflux for 5 hours. Insoluble material was removed by filtration through a cotton swab clogged pipette. The filtrate was concentrated in vacuo to give a residue, which was purified by reverse phase HPLC (YMC ODS-H80) eluted with 50% aqueous CH ₃ CN to give compound 4 (1.5 mg, 30%).

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 8.31 (1H, d, J = 2.4 Hz, H-4), 5.48 (1H, m, H-10), 5.22 (1H, m, H-9), 3.82 (3H, s, OCH ₃ ), 3.71 (3H, s, OCH ₃ ), 3.13 (1H, m, H-3), 2.64 (1H, dd, J = 16.0, 6.0 Hz, H-2b), 2.582 .51 (1H, m, H-8b), 2.49 (1H, dd, J = 16.0, 8.4 Hz, H-2a), 2.35 (1H, m, H-8a), 2.25 (1H, m, H-7 ), 2.04 (2H, m, H-11), 0.94 (3H, t, J = 7.6 Hz, H-12) FABMS m / z 281 [M + H] ⁺ HRFABMS m / z 281.1400 [M + H] ⁺ .

Example 8 4,5-Didehydrojasmonic acid (5)

To a solution of Compound 2 (8 mg) dissolved in acetone (0.1 mL) was added phosphate buffer (pH 8.0, 0.9 mL) and swine liver esterase (12 L) at ambient temperature and the mixture was stirred overnight. The solution was then acidified to dilute HCl solution to pH 4 and the mixture was extracted with EtOAc. The organic extract was dried over MgSO ₄ , filtered and evaporated to give a residue, which was purified by reverse phase HPLC (YMC ODS-H80) eluted with 6 5 % aqueous CH ₃ CN to give compound 5 (6 mg, 80%). Got.

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.63 (1H, dd, J = 6.0, 2.4 Hz, H-4), 6.20 (1H, dd, J = 6.0, 2.0 Hz, H-5), 5.45 ( 1H, m, H-10), 5.23 (1H, m, H-9), 2.99 (1H, m, H-3), 2.63 (1H, dd, J = 15.6, 6.8 Hz, H-2b), 2.552 .50 (1H, m, H-8b), 2.49 (1H, dd, J = 15.6, 8.4 Hz, H-2a), 2.30 (1H, m, H-8a), 2.09 (1H, m, H-7 ), 2.03 (2H, m, H-11), 0.93 (3H, t, J = 7.2 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 210.3 (C-6), 177.0 (C-1), 165.2 (C-4), 134.6 (C-10), 133.9 (C-5), 124.2 (C-9), 50.9 (C-7), 42.9 (C-3), 37.9 (C-2), 27.6 (C-8), 20.5 (C-11), 14.1 (C-12) FABMS m / z 209 [M + H] ⁺ HRFABMS m / z 209.1174 [M + H] ⁺ .

Example 9 Methyl 5-chloro-4,5-didihydrojasmonate (7)

30% hydrogen peroxide solution (50 L) was added to the stirred methanolic solution at 10 ° C. A few drops of 10% KOH (12 L) was added to the resulting mixture and stirred at room temperature for 12 hours. The reaction mixture was neutralized with 1N HCl and concentrated by evaporation with methanol. The residual oil was dissolved in EtOAc, washed with water, dried over MgSO ₄ and concentrated in vacuo to give epoxide compound 2a (20 mg). The epoxide compound was used without purification for the next reaction. A solution of compound 2a (5 mg) dissolved in acetic acid containing 1M HCl and CHCl ₃ was stirred at room temperature for 12 hours. The reaction mixture was diluted with CH ₂ Cl ₂ and washed three times with water. After working according to the standard method, the residue was purified using reverse phase HPLC (YMC ODS-H80) eluted with 65% aqueous CH ₃ CN to give compound 7 (2.0 mg, 38%).

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.52 (1H, d, J = 2.4 Hz, H-4), 5.48 (1H, m, H-10), 5.21 (1H, m, H-9), 3.70 (3H, s, OCH ₃ ), 2.96 (1H, m, H-3), 2.60 (1H, dd, J = 16.0, 6.4 Hz, H-2b), 2.562.51 (1H, m, H-8b ), 2.45 (1H, dd, J = 16.0, 7.6 Hz, H-2a), 2.35 (1H, m, H-8a), 2.22 (1H, m, H-7), 2.04 (2H, m, H- 11), 0.94 (3H, t, J = 7.6 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 201.4 (C-6), 171.4 (C-1), 158.1 (C-4), 135.7 (C-5), 135.2 (C-10), 123.6 (C -9), 52.0 (OCH ₃ ), 50.2 (C-7), 40.4 (C-3), 37.9 (C-2), 27.8 (C-8), 20.6 (C-11), 14.1 (C-12 ) FABMS m / z 257 [M + H] ⁺ HRFABMS m / z 257.0937 [M + H] ⁺ .

Example 10 5-Chloro-4,5-didehydrojasmonic acid (8)

Compound 8 (0.8 mg, 16%) was prepared in the same manner as Compound 7.

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz) δ 7.54 (1H, d, J = 2.8 Hz, H-4), 5.50 (1H, m, H-10), 5.23 (1H, m, H-9), 2.98 (1H, m, H-3), 2.68 (1H, dd, J = 16.0, 6.4 Hz, H-2b), 2.592.52 (1H, m, H-8b), 2.50 (1H, dd, J = 16.0 , 7.6 Hz, H-2a), 2.37 (1H, m, H-8a), 2.25 (1H, m, H-7), 2.06 (2H, m, H-11), 0.94 (3H, t, J = 7.6 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 201.5 (C-6), 176.6 (C-1), 157.9 (C-4), 135.9 (C-5), 135.3 (C-10), 123.2 (C -9), 50.2 (C-7), 40.1 (C-3), 37.7 (C-2), 27.8 (C-8), 20.6 (C-11), 14.1 (C-12) FABMS m / z 243 [M + H] ⁺ HRFABMS m / z 243.0765 [M + H] ⁺ .

Example 11 Methyl 5-bromo-4,5-didehydrojasmonate (9)

To a mixture of compound 2a (5 mg) and LiBr (3 equiv) dissolved in CH ₂ Cl ₂ was added silica gel (200 mg / mmol). The resulting suspension was shaken briefly at room temperature, evaporated to dryness. After holding at room temperature for 24 hours, silica gel was removed by filtration and the crude product was purified using reverse phase HPLC (YMC ODS-H80) eluted with 65% aqueous CH ₃ CN to give compound 9 (1.0 mg, 16%). Got.

Yellow oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.72 (1H, d, J = 2.8 Hz, H-4), 5.45 (1H, m, H-10), 5.23 (1H, m, H-9), 3.70 (3H, s, OCH ₃ ), 2.99 (1H, m, H-3), 2.60 (1H, dd, J = 16.0, 6.0 Hz, H-2b), 2.562.51 (1H, m, H-8b ), 2.45 (1H, dd, J = 16.0, 8.4 Hz, H-2a), 2.36 (1H, m, H-8a), 2.22 (1H, m, H-7), 2.03 (2H, m, H- 11), 0.94 (3H, t, J = 7.6 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 202.0 (C-6), 171.4 (C-1), 162.6 (C-4), 135.2 (C-10), 125.8 (C-5), 123.6 (C -9), 51.9 (OCH ₃ ), 49.8 (C-7), 42.4 (C-3), 37.8 (C-2), 27.9 (C-8), 20.6 (C-11), 14.1 (C-12 ) FABMS m / z 301/303 [M + H] ⁺ .

Example 12 Methyl 5-iodo-4,5-didehydrojasmonate (10)

Compound 10 (1.5 mg, 21%) was obtained by treating Compound 2a (5 mg), LiI (3 equivalents) and silica gel (200 mg / mmol) dissolved in CH ₂ Cl ₂ in the same manner as in the preparation of Compound 9. .

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.96 (1H, d, J = 2.4 Hz, H-4), 5.48 (1H, m, H-10), 5.20 (1H, m, H-9), 3.70 (3H, s, OCH ₃ ), 3.03 (1H, m, H-3), 2.58 (1H, dd, J = 16.0, 6.4 Hz, H-2b), 2.542.50 (1H, m, H-8b ), 2.45 (1H, dd, J = 16.0, 8.8 Hz, H-2a), 2.34 (1H, m, H-8a), 2.19 (1H, m, H-7), 2.04 (2H, m, H- 11), 0.94 (3H, t, J = 7.6 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 204.0 (C-6), 171.4 (C-1), 170.3 (C-4), 135.1 (C-10), 123.7 (C-9), 102.6 (C -5), 51.9 (OCH ₃ ), 48.5 (C-7), 45.3 (C-3), 37.8 (C-2), 27.9 (C-8), 20.6 (C-11), 14.1 (C-12 ) FABMS m / z 349 [M + H] ⁺ HRFABMS m / z 349.0314 [M + H] ⁺ .

Example 13 Methyl 4,5-didihydro-dihydrojasmonate (11)

Treatment of methyl dihydrozasmonate (50 mg) obtained by hydrogenation of IBX (2 equivalents) and TFA (0.3 equivalents) dissolved in DMSO under a palladium-charcoal (10% Pd-C) catalyst was carried out in the same manner as in the preparation of Compound 2. Compound 11 (16 mg, 32%) was obtained.

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.60 (1H, dd, J = 5.6, 2.4 Hz, H-4), 6.14 (1H, dd, J = 5.6, 2.4 Hz, H-5), 3.70 ( 3H, s, OCH ₃ ), 3.00 (1H, m, H-3), 2.56 (1H, dd, J = 16.0, 7.2 Hz, H-2b), 2.44 (1H, dd, J = 16.0, 8.4 Hz, H-2a), 1.98 (1H, m, H-7), 1.70 (1H, m, H-8b), 1.45 (1H, m, H-8a), 1.29 (6H, m, H-9, H- 10, and H-11), 0.85 (3H, t, J = 6.4 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 211.0 (C-6), 171.9 (C-1), 165.1 (C-4), 133.7 (C-5), 51.8 (OCH ₃ ), 51.2 (C- 7), 43.9 (C-3), 38.4 (C-2), 31.9 (C-10), 30.7 (C-9), 26.5 (C-8), 22.4 (C-11), 14.0 (C-12 ) FABMS m / z 225 [M + H] ⁺ HRFABMS m / z 225.1500 [M + H] ⁺ .

<Example 14> 4,5-didihydro-dihydrojasmonic acid (4,5-Didehydro-dihydrojasmonic acid; 12)

Compound 12 (6 mg) by treating Compound 11 (8 mg) and Porcine Liver Esterase (12 L) in the same manner as in the preparation of Compound 5 in a mixture of acetone (0.1 mL) and phosphate buffer (pH 8.0, 0.9 mL) , 80%).

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.64 (1H, dd, J = 5.2, 2.4 Hz, H-4), 6.18 (1H, dd, J = 5.2, 2.4 Hz, H-5), 3.02 ( 1H, m, H-3), 2.63 (1H, dd, J = 16.0, 6.4 Hz, H-2b), 2.49 (1H, dd, J = 16.0, 8.8 Hz, H-2a), 2.03 (1H, m , H-7), 1.73 (1H, m, H-8b), 1.47 (1H, m, H-8a), 1.30 (6H, m, H-9, H-10, and H-11), 0.85 ( 3H, t, J = 6.8 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 211.1 (C-6), 177.1 (C-1), 165.0 (C-4), 133.9 (C-5), 51.2 (C-7), 43.6 (C -3), 38.2 (C-2), 31.9 (C-10), 30.7 (C-9), 26.5 (C-8), 22.4 (C-11), 14.0 (C-12) FABMS m / z 211 [M + H] ⁺ HRFABMS m / z 211.1349 [M + H] ⁺ .

<Example 15> Methyl 5-chloro-4,5-didihydro-dihydrojasmonate (Methyl 5-chloro-4,5-didehydro-dihydrojasmonate; 13)

Compound 11a (5 mg) obtained by epoxidizing compound 11 in acetic acid including 1M HCl and CHCl ₃ was prepared in the same manner as in the preparation of compound 7, to obtain Compound 13 (2.1 mg, 40%).

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.53 (1H, d, J = 2.4 Hz, H-4), 3.71 (3H, s, OCH ₃ ), 2.99 (1H, m, H-3), 2.60 (1H, dd, J = 16.0, 6.0 Hz, H-2b), 2.46 (1H, dd, J = 16.0, 8.8 Hz, H-2a), 2.14 (1H, m, H-7), 1.75 (1H, m, H-8b), 1.53 (1H, m, H-8a), 1.29 (6H, m, H-9, H-10, and H-11), 0.86 (3H, t, J = 6.8 Hz, H -12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 202.0 (C-6), 171.5 (C-1), 157.9 (C-4), 135.7 (C-5), 52.0 (OCH ₃ ), 50.4 (C- 7), 41.2 (C-3), 38.2 (C-2), 31.8 (C-10), 30.7 (C-9), 26.3 (C-8), 22.4 (C-11), 14.0 (C-12 ) FABMS m / z 259 [M + H] ⁺ HRFABMS m / z 259.1103 [M + H] ⁺ .

Example 16 5-Chloro-4,5-didehydro-dihydrojasmonic acid (14)

Compound 14 (0.7 mg, 14%) was prepared in the same manner as in compound 13.

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.55 (1H, d, J = 2.4 Hz, H-4), 2.99 (1H, m, H-3), 2.68 (1H, dd, J = 16.4, 6.0 Hz, H-2b), 2.51 (1H, dd, J = 16.4, 9.6 Hz, H-2a), 2.17 (1H, m, H-7), 1.76 (1H, m, H-8b), 1.55 (1H , m, H-8a), 1.31 (6H, m, H-9, H-10, and H-11), 0.87 (3H, t, J = 6.8 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 202.1 (C-6), 176.7 (C-1), 157.7 (C-4), 135.5 (C-5), 50.4 (C-7), 40.9 (C -3), 38.0 (C-2), 31.8 (C-10), 30.7 (C-9), 26.3 (C-8), 22.4 (C-11), 14.0 (C-12); FABMS m / z 245 [M + H] ⁺ HRFABMS m / z 245.0961 [M + H] ⁺ .

Example 17 Methyl 5-bromo-4,5-didehydro-dihydrojasmonate (15)

Compound 15 (1.1 mg, 18%) was obtained by treating compound 11 a (5 mg), LiI (3 equivalents) and silica gel (200 mg / mmol) in CH ₂ Cl ₂ in the same manner as in the preparation of compound 9.

Yellow oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.73 (1H, d, J = 2.8 Hz, H-4), 3.72 (3H, s, OCH ₃ ), 2.99 (1H, m, H-3), 2.60 (1H, dd, J = 16.0, 6.4 Hz, H-2b), 2.46 (1H, dd, J = 16.0, 8.8 Hz, H-2a), 2.14 (1H, m, H-7), 1.76 (1H, m, H-8b), 1.50 (1H, m, H-8a), 1.29 (6H, m, H-9, H-10, and H-11), 0.86 (3H, t, J = 6.8 Hz, H -12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 202.0 (C-6), 171.4 (C-1), 162.3 (C-4), 125.6 (C-5), 52.0 (OCH ₃ ), 50.0 (C- 7), 43.2 (C-3), 38.0 (C-2), 31.8 (C-10), 30.9 (C-9), 26.3 (C-8), 22.4 (C-11), 14.0 (C-12 ) FABMS m / z 303/305 [M + H] ⁺ .

Example 18 Methyl 5-iodo-4,5-didihydro-dihydrojasmonate (Methyl 5-iodo-4,5-didehydro-dihydrojasmonate; 16)

Compound 16 (2.1 mg, 30%) was obtained by treating compound 11 a (5 mg), LiI (3 equivalents) and silica gel (200 mg / mmol) in CH ₂ Cl ₂ in the same manner as in the preparation of compound 9.

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.97 (1H, d, J = 2.4 Hz, H-4), 3.71 (3H, s, OCH ₃ ), 3.05 (1H, m, H-3), 2.58 (1H, dd, J = 15.6, 5.6 Hz, H-2b), 2.45 (1H, dd, J = 15.6, 8.8 Hz, H-2a), 2.14 (1H, m, H-7), 1.73 (1H, m, H-8b), 1.55 (1H, m, H-8a), 1.28 (6H, m, H-9, H-10, and H-11), 0.86 (3H, t, J = 6.8 Hz, H -12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 204.0 (C-6), 171.4 (C-1), 170.0 (C-4), 102.7 (C-5), 52.0 (OCH ₃ ), 48.7 (C- 7), 46.1 (C-3), 38.0 (C-2), 31.8 (C-10), 30.9 (C-9), 26.3 (C-8), 22.4 (C-11), 14.0 (C-12 ) FABMS m / z 351 [M + H] ⁺ HRFABMS m / z 351.0450 [M + H] ⁺ .

&Lt; Example 19 > N Isopropyl-4,5-didihydrozasmonylamide ( N -Isopropyl-4,5-didehydrojasmonylamide; 17)

To a mixture of compound 5 (2 mg) dissolved in EtOAc and triethylamine (TEA, 2 equiv) 2- (1H-benzotriazol-1 yl) -1,2,3,3-tetramethyluronium tetrafluoro Borate (TBTU, 1 equiv) was added. After stirring for 1 hour at room temperature, isopropylamine (2 equiv) was added and the reaction mixture was stirred for 12 hours. The mixture was washed with water, dried and purified using reverse phase HPLC (YMC ODS-H80) eluted with 65% water soluble CH ₃ CN to give compound 17 (1.5 mg, 63%).

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.65 (1H, dd, J = 5.6, 2.8 Hz, H-4), 6.14 (1H, dd, J = 5.6, 2.4 Hz, H-5), 5.45 ( 1H, m, H-10), 5.26 (1H, m, H-9), 4.07 (1H, septet, J = 6.4 Hz, -CH (CH ₃ ) ₂ ), 3.08 (1H, m, H-3) , 2.50 (1H, m, H-8b), 2.35 (1H, dd, J = 14.4, 6.8 Hz, H-2b), 2.29 (1H, m, H-8a), 2.22 (1H, dd, J = 14.4 , 8.4 Hz, H-2a), 2.05 (3H, m, H-7 and H-11), 1.14 (6H, d, J = 6.8 Hz,-(CH ₃ ) ₂ ), 0.94 (3H, t, J = 7.2 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 210.6 (C-6), 169.3 (C-1), 166.4 (C-4), 134.4 (C-10), 133.4 (C-5), 124.5 (C -9), 51.1 (C-7), 43.9 (C-3), 41.5 (C-2), 40.9 (-CH (CH ₃ ) ₂ ), 27.8 (C-8), 22.8 (-(CH ₃ ) ₂ ), 20.6 (C-11), 14.2 (C-12) FABMS m / z 250 [M + H] ⁺ HRFABMS m / z 250.1798 [M + H] ⁺ .

Example 20 N -Hydroxyethyl-4,5-didihydrozasmonylamide ( N -Hydroxyethyl-4,5-didehydrojasmonylamide; 18)

Compound 18 (1.3 mg, 54%) was obtained by treating compound 5 (2 mg), TBTU (1 equiv), TEA (2 equiv) and ethanolamine (2 equiv) in EtOAc in the same manner as in the preparation of compound 17. .

Yellow oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.66 (1H, dd, J = 5.6, 2.4 Hz, H-4), 6.15 (1H, dd, J = 5.6, 2.0 Hz, H-5), 5.89 ( 1H, brs, NH), 5.46 (1H, m, H-10), 5.26 (1H, m, H-9), 3.73 (2H, t, J = 5.6 Hz, -CH ₂ CH ₂ OH), 3.44 ( 2H, quint, J = 6.0 Hz, -CH ₂ OH), 3.08 (1H, m, H-3), 2.50 (1H, m, H-8b), 2.46 (1H, dd, J = 14.8, 6.8 Hz, H-2b), 2.30 (2H, m, H-2a and H-8a), 2.06 (3H, m, H-7 and H-11), 0.94 (3H, t, J = 7.6 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 210.6 (C-6), 171.3 (C-1), 166.2 (C-4), 134.5 (C-10), 133.5 (C-5), 124.5 (C -9), 62.2 (-CH ₂ OH), 51.1 (C-7), 43.8 (C-3), 42.2 (C-2), 40.5 (-CH ₂ CH ₂ OH), 27.8 (C-8), 20.6 (C-11), 14.2 (C-12) FABMS m / z 252 [M + H] ⁺ HRFABMS m / z 252.1588 [M + H] ⁺ .

&Lt; Example 21 > N Isopropyl-4,5-didihydro-dihydrozasmonylamide ( N -Isopropyl-4,5-didehydro-dihydrojasmonylamide; 19)

Compound 19 (1.5 mg, 63%) was treated with EtOAc 12 (2 mg), TBTU (1 equiv), TEA (2 equiv) and isopropylamine (2 equiv) in the same manner as in the preparation of compound 17. Got it.

Colorless oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.63 (1H, dd, J = 5.2, 2.4 Hz, H-4), 6.12 (1H, dd, J = 5.2, 1.6 Hz, H-5), 5.29 ( 1H, brs, NH), 4.08 (1H, septet, J = 6.8 Hz, -CH (CH ₃ ) ₂ ), 3.11 (1H, m, H-3), 2.33 (1H, dd, J = 14.0, 6.8 Hz , H-2b), 2.22 (1H, dd, J = 14.0, 8.4 Hz, H-2a), 1.93 (1H, m, H-7), 1.68 (1H, m, H-8b), 1.47 (1H, m, H-8a), 1.30 (6H, m, H-9, H-10, and H-11), 1.15 (6H, d, J = 6.8 Hz,-(CH ₃ ) ₂ ), 0.85 (3H, t, J = 6.4 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 211.5 (C-6), 169.3 (C-1), 166.1 (C-4), 133.3 (C-5), 51.4 (C-7), 44.5 (C -3), 41.5 (C-2), 41.2 (-CH (CH ₃ ) ₂ ), 31.9 (C-10), 30.8 (C-9), 26.5 (C-8), 22.8 (-(CH ₃ ) ₂ ), 22.4 (C-11), 14.0 (C-12) FABMS m / z 252 [M + H] ⁺ HRFABMS m / z 252.1945 [M + H] ⁺ .

<Example 22> N -Hydroxyethyl-4,5-didihydro-dihydrozasmonylamide ( N -Hydroxyethyl-4,5-didehydro-dihydrojasmonylamide; 20)

Compound 12 (2 mg), TBTU (1 equiv), TEA (2 equiv) and ethanolamine (2 equiv) were treated in the same manner as in the preparation of compound 17 in EtOAc to obtain compound 20 (1.2 mg, 50%). .

Yellow oil; ¹ H NMR (CDCl ₃ , 400 MHz): δ 7.65 (1H, dd, J = 6.0, 2.4 Hz, H-4), 6.13 (1H, dd, J = 6.0, 2.0 Hz, H-5), 5.94 ( 1H, brs, NH), 3.73 (2H, t, J = 5.2 Hz, -CH ₂ CH ₂ OH), 3.58 (2H, quint, J = 6.0 Hz, -CH ₂ OH), 3.11 (1H, m, H -3), 2.44 (1H, dd, J = 14.8, 6.8 Hz, H-2b), 2.30 (1H, dd, J = 14.8, 8.4 Hz, H-2a), 1.96 (1H, m, H-7) , 1.70 (1H, m, H-8b), 1.48 (1H, m, H-8a), 1.30 (6H, m, H-9, H-10, and H-11), 0.85 (3H, t, J = 6.8 Hz, H-12)

¹³ C NMR (CDCl ₃ , 100 MHz): δ 211.4 (C-6), 171.4 (C-1), 165.9 (C-4), 133.6 (C-5), 62.2 (-CH ₂ OH), 51.4 ( C-7), 44.4 (C-3), 42.2 (C-2), 40.9 (-CH ₂ CH ₂ OH), 31.9 (C-10), 30.8 (C-9), 26.5 (C-8), 22.5 (C-11), 14.0 (C-12) FABMS m / z 254 [M + H] ⁺ HRFABMS m / z 254.1763 [M + H] ⁺ .

Experimental Example 1 Cell Culture and Cytotoxicity Analysis

Murine macrophage RAW 264.7 was purchased from ATCC (Rockville, MD) and DMEM supplemented with 10% (v / v) heat-active fetal bovine serum, streptomycin (100 g / mL) and penicillin (100 U / mL) (Dulbecco's modified Eagle's medium) was incubated at a temperature of 37 ℃ and 5% CO ₂ .

For cytotoxicity analysis, MTT (3- [4,5-dimethylthiazol-2yl] -2,5-diphenyl tetrazolium bromide) assay was used in cells cultured for 24 hours. After 4 hours of adding MTT to the cultured cells, the cultures were removed from the incubator and the formazan crystals were dissolved by adding DMSO. Metabolic activity was quantified by measuring absorbance at 540 nm.

Experimental Example 2 Inhibition of Inflammatory Mediator

The production of nitric oxide (NO) was measured according to a previously known method ( Planta Med . 66 , 358360, 2000 ) using a Greases reagent (Sigma, MO, USA). That is, LPS (1 g / mL) was used to stimulate RAW 264.7 cells and 100 L of supernatant with 100 L of grease reagent (0.1% naphthylethylenediamine dihydrochloride, 1% sulfanylamide, 2.5% H ₃ PO ₄ ). Mixed. This mixture was incubated for 10 minutes at room temperature by blocking light. Absorbance was measured at 540 nm using an ELISA reader (Amersham Pharmacia Biotech, UK, USA) and the results were compared with a titration curve using sodium nitrite as standard.

In addition, the inhibitory activity on the production of IL-6 and TNF-α of the jasmonate derivative of the present invention was examined according to a known method ( Eur. J. Pharmacol . 398 , 399407, 2000). Samples were dissolved in EtOH diluted with DMEM and the final concentration of this solvent was not greater than 0.1% in culture medium. Under these conditions, lysed solvents did not alter IL-6 and TNF-α production in RAW 264.7 cells.

Before stimulation with LPS (1 g / mL) and test material, RAW 264.7 cells were incubated for 18 hours in 24-well plates. LPS and test material were then added to the cultured cells and IL-6 and TNF-α assays were performed using a mouse ELISA kit (R & D Systems Inc., MN, USA).

As a result, the jasmonate derivatives prepared in Examples as shown in FIGS. 1 to 3 inhibited the production of inflammatory mediators such as NO, IL-6 and TNF-α superior to the known methyl jasmonate. . In addition, as shown in FIG. 4, the jasmonate derivatives prepared in Examples inhibited the production of inflammatory mediators such as NO, IL-6 and TNF-α superior to the natural anti-inflammatory prostaglandins.

Experimental Example 3 Toxicity Test

150 ㅁ 5 g of ICR-based mice (central experimental animals) were divided into three groups of three, respectively, and the compound 7 prepared in Example was intraperitoneally administered at a dose of 20 mg / kg, 10 mg / kg, and 1 mg / kg, respectively. Toxicity was observed for 24 hours.

As a result, no deaths were observed in all three groups, and no symptoms were observed in weight gain and feed intake.

Hereinafter, an example of the preparation of a pharmaceutical composition containing the jasmonate derivative of the present invention will be described, but the present invention is not intended to be limited thereto, but is intended to be described in detail.

Preparation Example 1 Preparation of Injection

10 mg of compound 7 prepared in Example, 3.0 mg of sodium metabisulfite, 0.8 mg of methylparaben, 0.1 mg of propylparaben, and sterile distilled water for injection were mixed and prepared to have a final volume of 2 ml by a conventional method. An injection was prepared by filling a 2 ml ampoule and sterilizing it.

&Lt; Formulation Example 2 > Preparation of tablet

Tablets were prepared by mixing 10 mg of compound 7 prepared in the Examples, 100 mg of lactose, 100 mg of starch, and magnesium stearate in appropriate amounts and tableting according to a conventional tablet preparation method.

Preparation Example 3 Preparation of Capsule

10 mg of compound 7 prepared in Example, 50 mg of lactose, 50 mg of starch, 2 mg of talc, and magnesium stearate were mixed and filled into gelatin capsules according to a conventional capsule preparation method to prepare capsules.

Preparation Example 4 Preparation of Health Food

0.5 mg of compound 7 prepared in the Example, a suitable amount of vitamin mixture (70 μg of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B 1, 0.15 mg of vitamin B 2, 0.5 mg of vitamin B 6, 0.2 μg of vitamin B 12, vitamin C 10 mg, biotin 10 μg, nicotinic acid amide 1.7 mg, folic acid 50 μg, calcium pantothenate 0.5 mg and mineral mixture (1.75 mg ferrous sulfate, 0.82 mg zinc oxide, 25.3 mg magnesium carbonate, 15 mg potassium phosphate) , 55 mg of dibasic calcium phosphate, 90 mg of potassium citrate, 100 mg of calcium carbonate, 24.8 mg of magnesium chloride) were mixed, and then granules were prepared and health food was prepared according to a conventional method.

1 to 3 show the production inhibitory activity of the inflammatory mediator of the jasmonate derivative according to the present invention,

4 compares the NO inhibitory activity of the jasmonate derivative according to the present invention with prostaglandin, an anti-inflammatory agent,

Figure 5 shows the structure-activity correlation of the jasmonate derivative according to the present invention,

Figure 6 illustrates a jasmonate derivative according to the present invention,

Figure 7 shows the manufacturing process of the jasmonate derivative according to the present invention.

Claims (6)

Jasmonate derivative represented by the following formula (1): [Formula 1]

R ₁ is OCH ₃ , R ₂ is halogen, The bond between C-4 and C-5 and C-9 and C-10 is a single bond or a double bond with a dashed line. delete The method of claim 1, wherein the derivative is methyl-5-chloro-4,5-didihydrojasmonate; Methyl-5-bromo-4,5-didihydrozasmonate; Methyl-5-chloro-4,5-didihydro-dihydrozasmonate; And methyl-5-bromo-4,5-didihydro-dihydrozasmonate, wherein the compound is any one of jasmonate derivatives. A pharmaceutical composition for preventing and treating inflammatory diseases containing a jasmonate derivative represented by Formula 1 as an active ingredient: [Formula 1]

R ₁ is OCH ₃ , R ₂ is halogen, The bond between C-4 and C-5 and C-9 and C-10 is a single bond or a double bond with a dashed line. A pharmaceutical composition for preventing and treating immune diseases, comprising as an active ingredient a jasmonate derivative represented by Formula 1 below: [Formula 1]

R ₁ is OCH ₃ , R ₂ is halogen, The bond between C-4 and C-5 and C-9 and C-10 is a single bond or a double bond with a dashed line. Health foods for improving inflammatory diseases or immune diseases containing jasmonate derivative represented by the following formula 1 as an active ingredient: [Formula 1]

R ₁ is OCH ₃ , R ₂ is halogen, The bond between C-4 and C-5 and C-9 and C-10 is a single bond or a double bond with a dashed line.

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