KR20120084504A - Pharmaceutical composition for preventing or treating inflammatory diseases comprising sinapic acid - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing sinapinic acid is provided to suppress oxidative stress due to LPS treatment or aging to treat inflammatory diseases. CONSTITUTION: A pharmaceutical composition for preventing or treating inflammatory diseases contains 0.0005-0.0015 weight parts of sinapinic acid as an active ingredient. The sinapinic acid is a PPAR-alpha agonist. The pharmaceutical composition is manufactured in the form of powder, granules, tablets, capsules, suspensions, emulsions, syrups, and or suppository.

Description

Pharmaceutical composition for preventing or treating inflammatory diseases comprising sinapic acid}

The present invention relates to a pharmaceutical composition for preventing or treating an inflammatory disease containing cinafinic acid, a peroxysome growth factor activating receptor-alpha (PPAR-α) agonist, as an active ingredient.

Increasing living standards and increasing metabolic disorders such as hyperlipidemia, diabetes, and obesity are on the rise, and these diseases have complex causes, including genetic and nutritional factors.

Peroxysome (Peroxisome) is one of the intracellular organelles that cause these metabolic abnormalities, and has been considered to play a role in the cellular function for a long time, but a lot of recent research has been in the control of cell proliferation / differentiation, control of inflammatory mediators It plays an important role and has also been reported to have a broad effect on the metabolism of oxygen, glucose, lipids and hormones. Through lipid metabolism and glucose metabolism, peroxysomes are known to play an important role in senescence and tumorigenesis through not only insulin sensitivity but also cell membrane and adipocyte formation and effects on oxidative stress.

Over the past decade, nuclear hormone receptors called peroxisome proliferator activated receptors (PPARs) have been reported to be good targets as pharmacological approaches for many diseases, and recent studies on PPARs, particularly PPAR-α, one of the subtypes, plays an important role in promoting the differentiation / proliferation of keratinocytes in the epidermis of the skin, promoting skin barrier formation through lipid metabolism and suppressing inflammation. Inhibition of inflammatory mediators and erythema production by ultraviolet rays are known.

Looking at the patents for PPAR mainly known activator for γ type, that is, patents for new substances and patents for the search method, use patents for diabetes, obesity and the like. For example, substituted 4-hydroxyphenylalkanoic acid derivatives having agonist activity against PPAR- [gamma] (Korean Patent No. 474202), novel compounds useful for the treatment of PPAR mediated diseases (Korean Patent No. 2005-0055790) No.), chiral oxazole-arylpropionic acid derivatives and their use as PPAR agonists (Korean Patent No. 2005-0055750), novel PPAR ligands that do not cause fluid retention, edema or congestive heart failure (Korean Patent No. 2005- 0055701), N -substituted- 1H -indole-5-propionic acid compounds (Korean Patent Publication No. 2005-0042809) and the like as PPAR agonists useful for treating diabetes.

On the other hand, cinafinic acid is a phenylpropanoid-based natural carboxylic acid represented by the following formula (1), and is known to affect the cell wall structure by forming a dimer on its own or together with ferulic acid.

[Formula 1]

However, there is no research on the medical use of cinafinic acid as a new PPAR-α agonist.

Accordingly, the present inventors have made efforts to discover a new PPAR-α agonist, and as a result, cinafinic acid suppresses oxidative stress generated during LPS treatment or aging, and upregulates PPAR-α to activate NF by oxidative stress. In addition to inhibiting -κB, the present invention was completed by discovering that Apo A1 is increased to increase HDL.

An object of the present invention to provide a pharmaceutical composition for the prevention or treatment of inflammatory diseases containing cinafinic acid as an active ingredient.

In addition, another object of the present invention to provide a health food for improving inflammatory diseases containing cinnafinic acid as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases containing cinafinic acid as an active ingredient.

In particular, the cinafinic acid is a peroxysome growth factor activating receptor-alpha (PPAR-α) agonist, which inhibits oxidative stress caused by LPS in LPS-treated mice, and upregulates PPAR-α to oxidative stress. In addition to inhibiting the activated NF-κB, Apo A1 can be increased to increase HDL. In addition, the cinafinic acid inhibits the oxidative stress generated during the aging process, and upregulates PPAR-α to inhibit NF-κB activated by the oxidative stress, as well as increase Apo A1 to increase HDL. .

The inflammatory disease is any one selected from the group consisting of inflammatory vascular disease, allergic disease, inflammatory bowel disease, systemic lupus erythematosus, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, pancreatitis and nephritis The inflammatory vascular disease may be atherosclerosis.

In addition, the allergic disease may be any one selected from the group consisting of hypersensitivity, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis and urticaria.

The pharmaceutical composition according to the present invention preferably contains 0.0005 to 0.0015 parts by weight of cinafinic acid based on 100 parts by weight of the total pharmaceutical composition. If cinafinic acid is contained in a small amount out of the above content range, it is difficult to obtain the desired anti-inflammatory activity, and containing an excessive amount will slow down the treatment and prevention effect of inflammatory diseases compared to the dose, and may cause side effects, and economical This may cause problems.

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 .

In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is 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 sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The amount of cinafinic acid, which is an active ingredient of the pharmaceutical composition according to the present invention, may vary depending on the age, sex, and weight of the patient, but preferably 0.5 to 5 mg / kg, preferably 1 to 3 mg / kg once daily. can do.

In addition, the dosage of cinafinic acid 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 may be administered to various mammals such as mice, mice, livestock, humans, and the like. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrabronchial inhalation, intrauterine dural or intracerebroventricular injection.

The cinafinic acid has a 50% lethal amount (LD50) of 2 g / kg or more, the stability of which can be used in the pharmaceutical composition of the present invention.

In addition, the present invention provides a health food for improving inflammatory disease containing cinnafinic acid as an active ingredient.

The health food may be provided in the form of powder, granules, tablets, capsules, syrups or beverages.

The health food is used with other food or food additives in addition to cinnamic acid, and may be appropriately 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 cinafinic acid contained in the health food may be used according to the effective dose of the pharmaceutical composition, but may be less than the above range for long term intake for health and hygiene purposes or health control purposes. However, since the active ingredient has no problem in terms of safety, it is evident that it can be used in an amount above the above range.

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.

Cinafinic acid according to the present invention is a peroxysome growth factor activating receptor-alpha (PPAR-α) agonist, which inhibits oxidative stress generated by LPS treatment or aging process, and upregulates PPAR-α to oxidative stress. In addition to inhibiting the activated NF-κB, Apo A1 is increased to increase HDL may be useful for treating or preventing various inflammatory diseases including atherosclerosis.

1 is to examine the inhibitory effect of cinnafinic acid on ROS generated in SIN-1 treated vascular endothelial cells,
Figure 2 examines the inhibitory effect of cinnafinic acid on peroxynitrite generated in SIN-1 treated vascular endothelial cells,
Figure 3 examines the effect of increasing the PPARα activity according to the treatment with cinnafinic acid in vascular endothelial cells,
4 shows a molecular docking model between cinafinic acid and PPARα receptors,
Figure 5 examines the inhibitory effect of cinnafinic acid on ROS generated in LPS-treated mice,
Figure 6 examines the inhibitory effect of cinafinic acid on MDA and HAE generated in LPS treated mice,
Figure 7 examines the inhibitory effect of cinafinic acid on NF-κB translocation in LPS treated mice,
8 is a review of the effect of cinafinic acid on Apo A1 and PPARα expression in LPS treated mice,
Figure 9 examines the effect of cinafinic acid on HDL-cholesterol levels in LPS treated mice,
Figure 10 examines the effect of cinnafinic acid on TG levels in serum of LPS treated mice,
Figure 11 examines the inhibitory effect of cinnafinic acid on ROS generated in aging rats.
12 is to examine the inhibitory effect of cinnafinic acid on NO generated in aging rats.
13 is a review of the effects of cinafinic acid on NF-κB translocation and Apo A1 and PPARα expression in senescent rats.
Figure 14 examines the effect of cinafinic acid on HDL-cholesterol levels in aging rats.

Hereinafter, the present invention will be described in more detail with reference to the following Examples and Experimental Examples. However, these examples and the like are merely illustrative of the present invention, the contents of the present invention is not limited to these examples and the like.

≪ Example 1 > in Vitro  Experiment

1. Preparation of Vascular Endothelial Cells (YPEN-1)

YPEN-1 cells (rat prostatic endothelial cell line) were obtained from American Type Culture Collection, Manassas, VA, USA, which cells were 2 mM L-glutamine, 100 mg / ml streptomycin, 2.5 mg / L amphotericin B. And cultured using DMEM (Dulbecco's Modified Eagle Medium, Nissui, Tokyo, Japan) containing 5% inactivated fetal bovine serum (FBS). The cells were also maintained at 37 ° C. under the same conditions as a humid atmosphere containing 5% CO ₂ and 95% air. And 5% FBS was not added was used as a serum-free medium (SFM, serum-free medium). Cell lines were maintained by subculture every two days in a 100 mm plastic flask (Corning Co., New York, USA).

2. ROS measurement

It was measured by DCFDA (2 ', 7'-dichlorodihydrofluorescein diacetate) assay according to a known method (Chem Res Toxicol. 5: 227-231, 1992). That is, 12.5 mM DCFDA dissolved in 99.9% ethanol and 600 U / ml esterase dissolved in tertiary distilled water were stored as stock solutions at -20 ° C. In the experiment, DCFH (2 'prepared by mixing 10 mM DCFDA and 6 U / ml esterase) was used. , 7'-dichlorodihydrofluorescein) solution was incubated at 22 ° C. for 20 minutes and then stored frozen in the dark until use. The fat-soluble DCFDA undergoes esterase or oxidative hydrolysis to deacetylate it into non-fluorescent DCFH, and DCFH is oxidized by active oxygen to become a highly fluorescent DCF (2 ', 7'-dichlorofluorescein), so the excitation wavelength 485 nm And it was measured by a fluorescence photometer (GENios, TECAN) at an emission wavelength of 530nm. As a source of active oxygen, 50 μM of SIN-1 (3-morpholinosydnonimine hydrochloride) was pretreated to vascular endothelial cells for 1 hour.

As a result, as shown in FIG. 1, it was confirmed that cinnafinic acid inhibits ROS generated in vascular endothelial cells in a concentration-dependent manner.

3. Peroxynitrate Measurement

Samples were taken in 96-well microplates and sodium phosphate buffer (pH 7.4) containing 90 mM NaCl, 5 mM KCl and 100 mM diethylenetriaminepenta acetic acid and 10 mM DHR 123 was added. And it was measured at excitation 500nm and emission 536nm using a fluorophotometer (GENios, TECAN). At this time, as the source of peroxynitrite, SIN-1 (3-morpholinosydnonimine hydrochloride) 50μM was used for 1 hour pretreatment to vascular endothelial cells.

As a result, as shown in FIG. 2, it was confirmed that cinafinic acid suppressed peroxynitrate generated in vascular endothelial cells in a concentration-dependent manner.

Therefore, it was found that the oxidative stress generated from vascular endothelial cells was effectively inhibited by cinnafinic acid.

4. Western Blot Analysis

Cell culture containing a certain amount of protein and gel loading buffer (0.125M Tris-HCl, pH 6.8, 4% SDS, 10% 2-mercaptoethanol, 0.2% bromine phenol blue) was mixed and then boiled for 5 minutes. Each sample (100 mg) was electrophoresed at 110V for 1 hour 3 minutes in 10-15% sodium dodecyl sulfate (SDS) -polyacrylamide mini-gel. Transfer to polyvinylidene difluoride (PVDF) membrane with Towbin buffer (25 mM Tris-Cl, 192 mM glycine, 20% MeOH) and wash the membrane with 5% skim milk to block nonspecific protein binding After stirring for 1 hour in a buffer (10 mM Tris-Cl, pH 7.5, 100 mM NaCl, 0.1% tween-20), washed once with washing buffer and reacted with each primary antibody for 5 hours at room temperature. Washed three times with washing buffer and reacted with horseradish-peroxidase-conjugated secondary antibody for 1 hour at room temperature. After washing four times with a washing buffer, the phosphorescence generated by adding the ECL detection reagent was exposed to the hyperfilm to develop a signal. Molecular weight was determined using pre-stained blue protein markers (Bio-Rad).

As a result, as shown in Figure 3 as the treatment of cinnafinic acid in vascular endothelial cells it was confirmed that the concentration-dependent increase in the activity of PPARα.

<Example 2> in Silico  Experiment-Molecular Docking Analysis

Molecular docking analysis was observed using the program (J Mol Biol, 261: 470-489, 1996). The PPARa receptor structure was constructed in 3D using information obtained from X-rays to obtain binding values that appear when the histone, serine, tyrosine residues of the active receptor and cinnamic acid bind.

 As a result, it was observed that cinnafinic acid binds to the PPARα receptor, which is data recorded through the protein database as shown in FIG. 4.

Therefore, oxidative stress is inhibited by the treatment of cinnafinic acid in vascular endothelial cells, and one of the reasons why the inflammatory response is suppressed was determined to be because PPARα is activated to regulate NF-κB activity.

Example 3 In LPS Treated Mice in Vivo  Experiment

1. Preparation of LPS-treated Inflammatory Response Mice

SPF male 6-week-old C57BL / 6 mice were received from Samtako and tested. C57BL / 6 mice were intraperitoneally injected with cinafinic acid 1, 3, and 9 mg / kg / day for 2 days, and 1 hour later were intraperitoneally injected with LPS 5 mg / kg to induce an inflammatory response. Six animals were divided into groups. After 1 hour, sacrifice was performed using kidney tissue.

2. Preparation of animal test tissue

The 300 mg tissue prepared above was prepared using 2 ml of homogenate buffer A (10 mM HEPES, pH 7.8, 10 mM KCl, 2 mM MgCl 2, 1 mM dithiothreitol (DTT), 0.1 mM MEDTA, 0.1 mM MPMSF, 1 mM pepstatin, 1 mM p- Aminobenzamidine). After 15 minutes, 125% of 10% Nonidet p40 (NP 40) was added and mixed for about 20 seconds, followed by centrifugation at 12,000 rpm for 5 minutes to use the supernatant as a cytoplasmic fraction. 50 ml of buffer C (50 mM HEPES, pH 7.8, 50 mM KCl, 300 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 0.1 mM PMSF, 10% (v / v) glycerol) was mixed with the remaining pellets for 30 minutes under a cold reaction. Centrifuge for 5 minutes at. Supernatant was used as nuclear fraction.

3. ROS measurement

DCFDA was dissolved in pure ethanol and stored in the dark at -70 ℃ at a concentration of 12.5mM. The solution required for the experiment was prepared using phosphate buffer (50 mM, pH 7.4). The change in fluorescence generated by adding 25 mM DCFDA at the final concentration to 250 ml tissue homogenate of final volume was measured with a fluorescence photometer (GENios, TECAN) for 30 minutes at excitation 485 nm and emission 530 nm.

As a result, as shown in Figure 5 ROS generated by LPS concentration-dependently reduced by the treatment with cinafinic acid.

4. Lipid Peroxide Measurement

The concentrations of representative lipid peroxides, malondialdehyde (MDA) and 4-hydroxyalkenal (HAE), were measured using a Biooxytech LPO-586 Assay Kit (Oxis Health, Inc., Portland, OR, USA). Reactions containing 100 ml tissue homogenate, 8.1% sodium dodecyl sulfate, 20% acetate buffer (pH 3.5) and 0.8% thiobarbituric acid (TBA) solution were mixed for 3 minutes and then treated at 37 ° C. for 60 minutes. Absorbance at 582 nm was expressed in nmol per mg protein.

As a result, MDA and HAE generated by LPS as shown in Figure 6 was reduced in a concentration-dependent manner by the treatment with cinafinic acid.

5. Western Blot Analysis

Tissue homogenate containing a certain amount of protein and gel loading buffer (0.125M Tris-HCl, pH 6.8, 4% SDS, 10% 2-mercaptoethanol, 0.2% bromphenol blue) were mixed and then boiled for 5 minutes. Each sample (100 mg) was electrophoresed at 110V for 1 hour 3 minutes in 10-15% sodium dodecyl sulfate (SDS) -polyacrylamide mini-gel. Transfer to polyvinylidene difluoride (PVDF) membrane with Towbin buffer (25 mM Tris-Cl, 192 mM glycine, 20% MeOH) and wash the membrane with 5% skim milk to block nonspecific protein binding After stirring for 1 hour in a buffer (10 mM Tris-Cl, pH 7.5, 100 mM NaCl, 0.1% tween-20), washed once with washing buffer and reacted with each primary antibody for 5 hours at room temperature. Washed three times with washing buffer and reacted with horseradish-peroxidase-conjugated secondary antibody for 1 hour at room temperature. After washing four times with a washing buffer, the phosphorescence generated by adding the ECL detection reagent was exposed to the hyperfilm to develop a signal. Molecular weight was determined using pre-stained blue protein markers (Bio-Rad).

As a result of examining whether or not cinnafinic acid reduced NF-κB activated by LPS, as shown in FIG. 7, NF-κB activated by LPS was concentration-dependently reduced by cinnafinic acid treatment. Therefore, it was found that when PPARα is activated by cinnafinic acid treatment, NF-κB is down-regulated to inhibit the expression of inflammation-related genes.

In addition, as a result of examining whether cinafinic acid increases Apo A1 expression due to PPARα activation, as shown in FIG. 8, expression of PPARα and Apo A1 was increased in a concentration-dependent manner by cinnafinic acid treatment.

6. HDL-Cholesterol Measurement

HDL concentration was measured using a quantitative kit (HDL-C555, Shinyang Diagnostics, Seoul, Korea) using the tungstophosphoric acid-Mg ²⁺ precipitation method. 200 ml of serum and 200 ml of precipitant were mixed well, and left at room temperature for 10 minutes or more, centrifuged at 300 rpm or more for 10 minutes, 50 ml of the supernatant was mixed with 3 ml of enzyme solution, warmed for 5 minutes in a 37 ℃ hot water bath, and absorbed at 550 nm. Was measured.

As a result, the concentration of HDL in the serum as shown in Figure 9 was increased concentration-dependently by the treatment with cinnafinic acid.

7. TG Measurement

TG was measured using an quantitative kit by enzyme method (Triglyzyme-V, Shinyang Diagnostics, Seoul, Korea). 20 ml of serum and 200 ml of color developing solution were mixed well and warmed in a 37 ° C. hot water bath for 5 minutes and absorbance was measured at 550 nm.

As a result, the concentration of TG in the serum as shown in Figure 10 was reduced concentration-dependently by the treatment with cinnafinic acid.

Example 4 In Aged Rats in Vivo  Experiment

1. Aging Rats and Tissue Preparation

SPF male 6-month-old and 22-month-old SD rats were tested in Samtako. Cinnamic acid was fed to rats at 1 mg / kg / day and 3 mg / kg / day for 10 days. The experimental group consisted of a 6-month-old cinafinic acid 1mg / kg diet group, a 6-month-old cinafinic acid 3mg / kg diet group, a 22-month-old cinafinic acid 1mg / kg diet group, and a 22-month-old cinafinic acid 3 mg / kg diet group. The diets were classified as 6 months old with dietary restriction and 22 months old with 40% dietary restriction for 4 weeks. Cinafinic acid was sacrificed for 10 days and sacrificed to test with kidney tissue. At this time, the tissue was prepared in the same manner as in Example 3.

6 months old 22 months old 6 months old
Dietary restriction 22 months old
Dietary restriction Cinafinic acid
1mg / kg Cinafinic acid
3mg / kg Weight change Initial weight (g) 502.5 ± 10.16 631.6 ± 27.36 570.5 ± 18.52 647.8 ± 31.97 670.2 ± 38.81 642.6 ± 22.26 Final weight (g) 542.3 ± 7.01 660.4 ± 24.49 522.2 ± 15.77 630.6 ± 11.36 671.2 ± 38.32 640.2 ± 45.23 Diet (g / day) 22.9 ± 0.46 27.7 ± 0.86 15.2 ± 0.28 15.6 ± 0.30 26.3 ± 2.06 24.9 ± 0.38

2. ROS measurement

As a result of measuring the ROS in the aging rats in the same manner as in Example 3, it was observed that ROS induced by the aging process is dependent on the concentration of cinnafinic acid in the group fed with cinnamic acid as shown in FIG. 11.

3. NO measurement

10 ml tissue homogenate and 140 ml of 50 mM phosphate buffer were mixed in a 96-well plate, and NO production was measured. NO activity was measured by fluorescence spectrophotometry at 495 nm exitation and 515 nm emission for 30 minutes using DAF-2. The amount of NO produced was obtained by dividing the change in fluorescence value per minute by the protein concentration.

As a result, as shown in FIG. 12, the NO induced by the aging process was observed to decrease depending on the concentration of cinnafinic acid in the group fed the cinnamic acid.

4. Western Blot Analysis

NF-κB and PPARα protein expression in nuclear extracts in senescent rats was analyzed in the same manner as in Example 3.

As a result, it was observed that NF-kB activated during the aging process also decreased depending on the concentration of cinafinic acid as shown in FIG. 13. In addition, cinafinic acid was observed to increase concentration-dependently Apo A1 expression due to PPARα activation.

5. HDL-Cholesterol Measurement

As a result of measuring HDL-cholesterol in aging rats in the same manner as in Example 3, it was observed that HDL was increased in serum depending on the concentration of cinnafinic acid.

Example 5 Toxicity Test

Male Balb / c mice were suspended in 0.5% methylcellulose solution in cinafinic acid in a single oral dose of 0.5 g / kg, 1 g / kg and 2 g / kg and examined for survival and body weight for 7 days.

After the administration, the mortality, clinical symptoms, and weight changes of the animals were observed. Hematological and hematological examinations were performed. Necropsy was performed to visually observe the abdominal and thoracic organ abnormalities.

As a result, no significant clinical symptoms or dead animals were noted in all animals, and no toxic changes were observed in weight changes, blood tests, blood biochemistry tests, and autopsy findings.

As a result, the cinafinic acid of the present invention does not show a change in toxicity in rats up to 2 g / kg, therefore, the oral administration intermediate dose (LD50) was determined to be a safe substance of more than 2 g / kg.

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

Preparation Example 1 Preparation of Powder

Powder was prepared by mixing 20 mg of cinnafinic acid, 100 mg of lactose and 10 mg of talc and filling into an airtight cloth.

&Lt; Formulation Example 2 > Preparation of tablet

Tablets were prepared by mixing 20 mg of cinnafinic acid, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate, followed by compression according to a conventional method for preparing tablets.

Preparation Example 3 Preparation of Capsule

Cinnamic acid 10 mg, corn starch 100 mg, lactose 100 mg and magnesium stearate 2mg were mixed and the above ingredients were mixed according to a conventional capsule preparation method and filled into gelatin capsules to prepare a capsule.

Preparation Example 4 Preparation of Injection

Cinnamic acid 10 mg, sterile distilled water for injection, and a pH adjusting agent were mixed, and then prepared in the above-described ingredient content per ampoule (2 ml) according to a conventional injection method.

Preparation Example 5 Preparation of Health Food

Cinnamic acid 1 mg, vitamin mixture (70 mg of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B, 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, 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 monophosphate, calcium diphosphate dibasic) 55 mg, potassium citrate 90 mg, calcium carbonate 100 mg, magnesium chloride 24.8 mg) were mixed, granules were prepared, and health food was prepared according to a conventional method.

Preparation Example 6 Preparation of Health Beverage

1 mg of cinnamic acid, 1000 mg of citric acid, 100 g of oligosaccharide, 2 g of plum concentrate, 1 g of taurine and purified water are added to make it total 900 ml. After stirring and heating at 85 ° C. for an hour, the resulting solution was collected by filtration into a sterilized 2 L container, sealed sterilized and then refrigerated.

Claims (7)

A pharmaceutical composition for preventing or treating inflammatory diseases containing cinnafinic acid as an active ingredient. The pharmaceutical composition for preventing or treating inflammatory diseases according to claim 1, wherein the cinafinic acid is a peroxysome growth factor activating receptor-alpha (PPAR-α) agonist. The method according to claim 1 or 2, wherein the inflammatory disease is inflammatory vascular disease, allergic disease, inflammatory bowel disease, systemic lupus erythematosus, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, pancreatitis and nephritis Inflammatory disease prevention or treatment pharmaceutical composition, characterized in that any one selected from the group consisting of. The pharmaceutical composition for preventing or treating inflammatory diseases according to claim 3, wherein the inflammatory disease is atherosclerosis. The method of claim 3, wherein the allergic disease is any one selected from the group consisting of hypersensitivity, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis and urticaria Or therapeutic pharmaceutical compositions. Health food for improving inflammatory disease containing cinnamic acid as an active ingredient. The health food for improving inflammatory disease according to claim 6, wherein the health food is a powder, granule, tablet, capsule, syrup or beverage.

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