KR101281108B1 - Lysophosphatidylethanolamine derivatives having anti-inflammatory activity or pharmaceutically acceptable salt thereof, preparation method thereof and anti-inflammatory composition containing the same as active ingredient - Google Patents

Abstract

The present invention relates to a lysophosphatidylethanolamine derivative having anti-inflammatory activity or a pharmaceutically acceptable salt thereof, a method for preparing the same, and an anti-inflammatory composition containing the same as an active ingredient, wherein the lysophosphatidylethanolamine derivative according to the present invention is oral Dosing inhibits zymosan A-induced peritonitis in vivo and is an inflammatory factor, lipid mediators (LTC ₄ , LTB ₄ ) or cytokines (TNF-α, IL-1 and IL-6) Inhibits the production of) and increases the production of anti-inflammatory cytokines (IL-10), thereby exhibiting excellent anti-inflammatory activity, which can be usefully used as an active ingredient in pharmaceutical compositions, health foods or cosmetic compositions for preventing and treating inflammatory diseases. Can be.

Description

Lysophosphatidylethanolamine derivatives having anti-inflammatory activity or pharmaceutically acceptable salts thereof, preparation methods thereof and anti-inflammatory compositions containing them as active ingredients anti-inflammatory composition containing the same as active ingredient}

The present invention relates to a lysophosphatidylethanolamine derivative having anti-inflammatory activity or a pharmaceutically acceptable salt thereof, a method for preparing the same, and an anti-inflammatory composition containing the same as an active ingredient.

Inflammatory reactions are associated with various inflammatory mediators and immune cells in local blood vessels and body fluids when they are damaged by tissues (cells) or by external infectious agents (bacteria, fungi, viruses, or various types of allergens). It has a series of complex physiological reactions, including substance secretion, fluid infiltration, cell migration and tissue destruction, as well as external symptoms such as erythema, edema, fever and pain. In normal cases, the inflammatory response removes external infectious agents and regenerates damaged tissues to restore the function of life.However, if the inflammatory response is excessive or persistent due to the absence of antigens or internal substances, it promotes mucosal damage. Some of the findings lead to diseases such as cancer development (Jonathan Cohen, Nature, 420, 885-891, 2002).

Inflammation causes a variety of biochemical phenomena in the living body, and especially the biosynthesis of nitric oxide synthase (NOS) and prostaglandin, enzymes that produce nitric oxide (NO). Related enzymes are known to play an important role in mediating the inflammatory response. Thus, NOS, an enzyme that produces NO from L-Arginine, or cyclooxygenase (COX), an enzyme involved in synthesizing prostaglandins from arachidonic acid, is a major factor in blocking inflammation. It is a goal.

Recent studies show that there are several types of NOS: brain NOS (bNOS) in the brain, neuronal NOS (nNOS) in the nervous system, and in the vascular system. Endothelial nitric oxide synthase (endothelial NOS; eNOS) is always expressed at a certain level in the body, and a small amount of NO produced by them plays an important role in maintaining normal homeostasis, such as inducing neurotransmission and vasodilation. On the other hand, NO, which is excessively generated by iNOS (induced NOS) induced by various cytokines or external stimulants, is known to cause cytotoxicity and various inflammatory reactions. Chronic inflammation is associated with an increase in iNOS activity. There is a study (Jon O. Lundberg, et al., Nature Reviews Drug Discovery, 2008). In addition, there are two types of isoforms of COX, while COX-1 is present in cells and synthesizes prostaglandins (PGs) necessary for cytoprotective action. It is known to increase rapidly and play an important role in causing an inflammatory response. Transcriptional inflammatory factors, including iNOS and COX-2, which enhance the degree of NO and PGs, are chronic diseases, including various sclerosis, parkinson's disease, Alzheimer's disease and colon cancer. (Bengt Samuelsson, et al., Pharmacological Reviews, 59, 207-224, 2007).

Lipopolysaccharide (LPS) is an externally secreted bacterial toxin, which stimulates cellular inflammatory responses, nitric oxide (NO), cytokines, tumor necrosis factor (TNF-α), Secretes a variety of inflammatory modulators, including prostaglandin E2 and eicosanoid modulators that promote inflammatory responses (Chen YC, et al., Biochem. Pharmacol., 61, 1417-). 1427, 2001). Nitric oxide synthase is classified into two groups. CNOS is consistently present in various cells (for example, neurons and endothelial cells), and its transcription level is increased by calcium-dependent calmodulin. It is controlled first. On the other hand, smooth muscle cells, macrophages, hepatocytes and astrocytes are induced in response to inflammatory cytokines and lipopolysaccharides. Activation of nitric oxide synthase promotes the formation of large amounts of nitric oxide, which plays an important role in diseases of various inflammatory diseases. Thus, nitric oxide production induced by nitric oxide synthase is an indicator of the extent of inflammation and an indicator of the progression of inflammation. Expression of specific cellular genes includes COX-2 and inducible nitric oxide synthase, as well as several inflammatory transcription factors such as interleukin-1, interleukin-2, interleukin-6 and tumor necrosis factor (Csaba Szabo, et al. , Nature Reviews Drug Discovery, 6, 662-680. 2007).

Recently, lipid-inducing mediators have been known to be involved in various inflammatory reactions (asthma, rheumatoid arthritis or enteritis) (Daleau P., J. Mol. Cell. Cardiol, 1999; 31: 1391-1401; Fuchs B, et al. ., Clin. Biochem 2005; 38: 925-933; Muralikrishna Adibhatla, R., et al., Free Radic. Biol. Med 2006; 40: 376-87; Triggiani M, et al., Biochim Biophys Acta. 1761 ( 11): 1289-300, 2006; Matsumoto, T., et al., Curr. Med. Chem. 14, 3209-3220, 2007; Shi Y, et al., Atherosclerosis 2007; 191: 54-62).

Although research on inflammatory mediators has been actively conducted, the involvement of lipid mediators in the inflammatory response has recently been established. For example, arachidonic acid liberated from phosphatidylcholine by the enzyme PLA2 is converted to prostaglandin (PG), leukotriene (LT) and lipoxin (LX), where prostaglandin and leukotriene are converted into Whereas it is a potent inflammatory mediator (Serhan CN, et al., Nat Immunol. 6: 1191-7, 2005; Farooqui AA, et al., J Neurochem. 101 (3): 577-99, 2007; Funk, CD, Science 294, 1871-5, 2001), rather, lipoxin has become known as a strong anti-inflammatory mediator (Schwab JM, et al., Curr Opin Pharmacol. 6 (4): 414-20, 2006; Kuhn H, et. al., Prog Lipid Res. 2006 Jul; 45 (4): 334-56.Epub 2006 M). That is, during inflammatory processes, the eicosanoids produced from arachidonic acid are converted to prostaglandins and leukotrienes, where 5-lipoxygenase is important for the formation of leukotriene and lipoxin. 12 / 15-lipoxygenase is important for the formation of lipoxin (Schwab JM, et al., Curr Opin Pharmacol. 6 (4): 414-20, 2006; Kuhn H, et. al., Prog Lipid Res. 2006 Jul; 45 (4): 334-56.Epub 2006 M). In addition, saturated lipid lysophosphatidylcholine (lysoPC) produced during PLA2 hydrolysis has been shown to be inflammatory or anti-inflammatory in the inflammatory response (Daleau P., J. Mol. Cell. Cardiol, 1999; 31: 1391-1401 Fuchs B, et al., Clin. Biochem 2005; 38: 925-933; Muralikrishna Adibhatla, R., et al., Free Radic. Biol.Med 2006; 40: 376-87; Shi Y, et al., Atherosclerosis 2007; 191: 54-62; Fuentes L, et al., Circ Res 2002, 91: 681-688). It has also been reported that the action of inflammatory lysophosphatidylcholine is the production of nitric oxide (NO) or reactive oxygen species (ROS) (Colles, SM et al., Journal of Lipid Research 2000; 41: 1188). -1198; Matsubara, M., et al., Atherosclerosis 2005; 178: 57-66; Takeshita S, et al., J. Atheroscler. Thromb 2000; 7: 238-246; Silliman CC, et al., J. Leukoc. Biol 2003; 73: 511-524; Cheon Ho Park, et al., Lysophosphatidylcholine exhibits a selective cytotoxicity, accompanied by ROS formation, in RAW 264.7 macrophages, Lipids, in press, 2009).

Recently, the effects of polyunsaturated lysoPCs containing polyunsaturated lipids on inflammatory responses in vivo have been reported. That is, it was found that arachidonoyl lysophosphatidylcholine (achidonoyl lysoPC) and docosahexaenoyl lysophosphatidylcholine (docosahexaenoyl lysoPC) have excellent anti-inflammatory effects on the inflammatory response in vivo (Nguyen DH et al, 2009; Nguyen DH et al. , 2010). Moreover, polyunsaturated lysophosphatidylcholine containing unsaturated lipids has been reported to be present in significant amounts in animals, such as linoleoyl lysophosphatidylcholine, arachidonoyl lyphophosphatidylcholine (arachidonoyl lysoPC) and docosahexaenoyl lysophosphatilis. docosahexaenoyl lysoPC) is known as plasma (Okita, M. et al., Int. J. Cancer. 1997, 71, 31-34; Croset, M. et al., Biochem. J. 2000, 345, 61- 67; Adachi, J. et al., Kobe.J. Med. Sci. 2006, 52, 127-140), in particular docosahexaenoyl lysophosphatidylcholine is one of the major lipids in the shark liver. (Chen, S., et al., J. Agric. Food. Chem. 2007, 55, 9670-9677). Moreover, choline oxide is present in the heart extract. Therefore, lysophosphatidylcholine containing unsaturated lipids is thought to be metabolized by oxidase, which has recently been demonstrated (Huang, LS et al., Arch. Biochem. Biophys. 2006, 455, 119-126; Huang, LS et. al., Lipids 2007, 42, 981-990; Huang, LS et al., J. Agric.Food.Chem. 2008, 56, 1224-1232. That is, lysophosphatidylcholines, including groups consisting of linoleoyl, arachidonoyl and docohexaenoyl, are reticulocyte 15-LOX or leukocyte 12 /. Good oxidation has been reported by 15-LOX enzymes (Huang, LS et al., Arch. Biochem. Biophys. 2006, 455, 119-126; Huang, LS et al., Lipids 2007, 42, 981-990; Huang, LS et al., J. Agric.Food.Chem. 2008, 56, 1224-1232). However, the physiological activity of unsaturated lipid-containing lysophosphatidylethanolamine derivatives is not known yet.

Accordingly, the present inventors have studied the physiological activity of the unsaturated lipid-containing lysophosphatidylethanolamine derivative, the lysophosphatidylethanolamine derivative is less toxic, in vivo anti-inflammatory effect, in particular when oral administration The present invention was completed by identifying that Zymosan A (zymosan A) -induced peritonitis was inhibited even at a low concentration, and showing that it can be used as an active ingredient of an anti-inflammatory composition.

An object of the present invention is to provide a lysophosphatidylethanolamine derivative having a anti-inflammatory activity or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a method for preparing the lysophosphatidylethanolamine derivative.

Still another object of the present invention is to provide an anti-inflammatory composition containing the lysophosphatidylethanolamine derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to achieve the above object, the present invention provides a lysophosphatidylethanolamine derivative or a pharmaceutically acceptable salt thereof having an anti-inflammatory activity represented by the following formula (1) to 4 having an anti-inflammatory activity.

[Formula 1]

[Formula 2]

(3)

[Formula 4]

The present invention also provides a method for preparing the lysophosphatidylethanolamine derivative.

Furthermore, the present invention provides a pharmaceutical composition for the prevention and treatment of inflammatory diseases containing the lysophosphatidylethanolamine derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also provides a health food for the prevention and improvement of inflammatory diseases containing the lysophosphatidylethanolamine derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

Furthermore, the present invention provides an anti-inflammatory skin external preparation containing the lysophosphatidylethanolamine derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also provides an anti-inflammatory cosmetic composition containing the lysophosphatidylethanolamine derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

The lysophosphatidylethanolamine derivative according to the present invention is orally administered to inhibit peritonitis induced by zymosan A in vivo and is an inflammatory factor, lipid mediators (LTC4, LTB4) or cytokines (TNF- It inhibits the production of α, IL-1 and IL-6) and increases the production of anti-inflammatory cytokines (IL-10), thereby exhibiting excellent anti-inflammatory activity, and therefore, pharmaceutical compositions, health foods or It can be usefully used as an active ingredient of a cosmetic composition.

1 shows LC / ESI- of a product resulting from oxidation using soybean 15-LOX catalyst of 2-ARA-lysoPtdEtn (A, B) or 2-DHA-lysoPtdEtn (C, D) according to one embodiment of the present invention. MS analysis result.
2 is a graph showing the effect of preventing Zymosan A induced plasma leakage on 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn according to an embodiment of the present invention.
FIG. 3 is a graph showing the anti-inflammatory effect of 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn on Zymosan A induced leukocyte extravasation according to an embodiment of the present invention.
4 is a graph showing the inhibitory effect on the formation of Zymosan A induced proinflammatory lipid mediators in mice of 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn according to an embodiment of the present invention.
FIG. 5 is a graph showing the effect of 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn on zymosan A-induced pro-inflammatory and anti-inflammatory cytokine formation according to one embodiment of the present invention.
6 is a graph showing the inhibitory effect of 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn on the formation of Zymosan A induced nitric oxide (NO) in mice according to an embodiment of the present invention.
7 is a graph showing the inhibitory effect of polyunsaturated acyl-lysophosphatidylethanolamine derivatives on Zymosan A-induced plasma efflux or leukocyte efflux in accordance with one embodiment of the present invention.
8 is a graph showing the dose dependent effect on the level of 15-HETE or LXA4 before and after alkaline hydrolysis of effusion from 2-ARA-lysoPtdEtn mice according to one embodiment of the present invention.
Figure 9 is a graph showing the induction of degradation of Zymosan A induced peritonitis by 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn according to an embodiment of the present invention.

Hereinafter, terms used in the present invention are defined.

The term "inflammation" as used herein refers to the defense response of biological tissues against local injury to the body. In other words, the inflammatory response is a biodefense reaction that attempts to recover the original state by removing the injury caused by the stimulus in response to various harmful stressors. Inflammatory stimuli include infection or chemical or physical stimuli. Bioconstituents involved in the inflammatory response include low-molecular or high-molecular chemicals such as free radicals, proteins, sugars, lipids, plasma, blood cells, blood vessels, and connective tissue. The process of inflammation is usually divided into two types, which can be divided into acute and chronic inflammation. Acute inflammation is a short-term reaction within a few days. Plasma components and blood cells are associated with the removal of foreign bodies with microcirculation. Chronic inflammation has a long duration, and tissue growth is seen.

As used herein, the term "prevention" means any action that delays the development of an inflammatory disease by administration of a composition of the present invention.

As used herein, the terms "treatment" and "improvement" refer to any action in which the symptoms of the disease improve or benefit altered by administration of the composition of the present invention.

As used herein, the term "administration" means providing a subject with any of the compositions of the present invention in any suitable manner.

Hereinafter, the present invention will be described in detail.

The present invention provides a lysophosphatidylethanolamine derivative represented by the following Chemical Formulas 1 to 4 or a pharmaceutically acceptable salt thereof.

Specific compounds of the lysophosphatidylethanolamine derivative represented by Formula 1-4 of the present invention are as follows:

1) 1-lyso-2-arachidonoyl-sn-glycero-3-phosphoethanolamine (2-ARA-lysoPtdEtn) (Formula 1);

2) 1-lyso-2-docosahexaenoyl-sn-glycero-3-phosphoethanolamine (2-DHA-lysoPtdEtn) (Formula 2);

3) 1-Liso-2-15 (S) -hydroxy-5,8,11,13-aicosapentaenoyl-sn-glycero-3-phosphoethanolamine (2- (15-HETE)- lysoPtdEtn) (Formula 3); And

4) 1-Liso-2-17 (S) -hydroxy-4,7,10,13,15,19-docosahexaenoyl-sn-glycero-3-phosphoethanolamine (2- (17 -HDHE) -lysoPtdEtn) (Formula 4).

The lysophosphatidylethanolamine derivative of the present invention can be used in the form of a pharmaceutically acceptable salt, and acid salts formed by the pharmaceutically acceptable free acid are useful as salts. The expression pharmaceutically acceptable salt is a concentration that has a relatively nontoxic and harmless effect on the patient, and any organic or inorganic addition of the compound of formula 1 in which the side effects caused by the salt do not compromise the beneficial efficacy of the compound of formula 1 Salt. Examples of the inorganic acid include hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid and phosphoric acid. Examples of the organic acid include citric acid, acetic acid, lactic acid, maleic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, maleic acid, succinic acid, tartaric acid, galacturonic acid, embic acid, glutamic acid, aspartic acid, oxalic acid, P-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid or malonic acid. These salts also include alkali metal salts (sodium salts, potassium salts, etc.) and alkaline earth metal salts (calcium salts, magnesium salts, etc.). For example, the acid addition salt may be selected from the group consisting of acetate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camylate, citrate, eddylate, Hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, malate, glucoside, gluconate, gluconate, glucuronate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / Hydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydroxyacetate, Lactate, stearate, succinate, tartrate, tosylate, trifluoroacetate Diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, zinc salts, and the like. Preferred is hydrochloride or trifluoroacetate.

In addition, the compounds of Formulas 1 to 4 of the present invention include all salts, hydrates, and solvates that can be prepared by conventional methods, as well as pharmaceutically acceptable salts.

The addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the compound of Formula 1 to 4 in a water miscible organic solvent such as acetone, methanol, ethanol, or acetonitrile and adding excess organic acid. Or by adding an aqueous acid solution of an inorganic acid and then precipitating or crystallizing. Subsequently, in this mixture, a solvent or an excess acid is evaporated and dried to obtain an additional salt, or the precipitated salt may be produced by suction filtration.

The present invention also provides a method for preparing a lysophosphatidylethanolamine derivative represented by Chemical Formulas 1 to 4.

Recipe 1

The compound of Formula 1 to 2 according to the present invention

Adding C18 (plasm) -PtEtn (1- (1Z-octadecenyl) -2-arachidonoyl-sn-glycero-3-phosphoethanolamine) to a mixed solvent of HCl, methanol and chloroform and stirring to hydrolyze (step 1);

It can be prepared by a method comprising the step of separating and purifying the hydrolysis product prepared in step 1 (step 2).

Specifically, in Step 1, C18 (plasm) -PtEtn (1- (1Z-octadecenyl) -2-arachidonoyl-sn-glycero-3-phosphoethanolamine) is added to a mixed solvent of HCl, methanol, and chloroform, followed by hydrolysis. Step. PtEtn is not limited in this step, but it is preferable to use 22: 6 PtEtn or 20: 4 PtEtn.

The hydrolyzed product may be separated and purified using extraction, thin layer chromatography (TLC), and the like, which are commonly used in the art, and then purified to 1-lyso-2-arachidonoyl-sn-glycerol. 3-phosphoethanolamine, 1-lyso-2-docosahexaenoyl-sn-glycero-3-phosphoethanolamine can be prepared.

Recipe 2

In addition, the compounds of Formulas 3 to 4 according to the present invention

Adding soybean lipoxygenase (soybean LOX-1) to a borax buffer containing lysophosphatidylethanolamine and reacting at room temperature to produce lysophosphatidylethanolamine peroxide (step 1); And

It can be prepared by a method comprising the step of reducing the addition of a reducing agent to the lysophosphatidylamine peroxide obtained in step 1 to produce a lysophosphatidylethanolamine hydroxide derivative (step 2).

First, step 1 is a step in which soybean lipoxygenase (soybean LOX-1) is added to borax buffer containing lysophosphatidylethanolamine and reacted at room temperature to produce lysophosphatidylcholine peroxide.

In the preparation method according to the present invention, the lysophosphatidylethanolamine is 1-lyso-2-arachidonoyl-sn-glycero-3-phosphoethanolamine prepared in Preparation Method 1, 1-lyso-2- Docosahexaenoyl-sn-glycero-3-phosphoethanolamine may be used, but is not limited thereto.

Specifically, soybean lipoxygenase is added to the borax buffer containing lysophosphatidylethanolamine and reacted at 20-30 ° C. for 20 minutes to 1 hour, whereby a part of the lysophosphatidylethanolamine is oxidized to lysophosphatidyl by lipoxygenase. Ethanolamine peroxide can be obtained.

Next, step 2 is a step of producing a lysophosphatidylethanolamine hydroxide derivative by reducing by adding a reducing agent to the lysophosphatidylethanolamine peroxide obtained in step 1 above.

In the manufacturing method according to the present invention, the reducing agent may use SnCl ₂ and the like, but is not limited thereto.

The lysophosphatidylethanolamine derivative produced by step 2 may be further purified by performing column chromatography.

Furthermore, the present invention provides a pharmaceutical composition for the prevention and treatment of inflammatory diseases containing the lysophosphatidylethanolamine derivative as an active ingredient.

At this time, the inflammatory disease is preferably any one selected from the group consisting of asthma, periodontitis, stomatitis, peritonitis, gastritis, enteritis, arthritis, nephritis, hepatitis and degenerative diseases, the group consisting of asthma, peritonitis, enteritis and rheumatoid arthritis It is more preferably any one selected from, and most preferably, but not limited to peritonitis.

In order to confirm the superior anti-inflammatory efficacy of the lysophosphatidylethanolamine derivatives according to the present invention, after oral administration of the lysophosphatidylethanol amine derivatives according to the present invention to mice, peritonitis is induced after a certain time using Zymosan A. Plasma outflow induced by Zymosan A, leukocyte outflow, inflammatory mediator lipids, inflammatory cytokines, anti-inflammatory cytokines, nitric oxide, leukocyte infiltration numbers, etc. were measured. In one group, concentration-dependent plasma outflow induced with Zymosan A was reduced (see FIGS . 2 and 7 ), leukocyte outflow was reduced (see FIGS . 3 and 8 ), and abdominal cavity such as LTC4 and LTB4 inflammation in the generation of lipid mediators was suppressed (see Fig. 4), a sense of abdominal significantly depending on the inflammatory cytokine is IL-1, IL-6, it produced a dose of TNF-α (See Fig. 5) and the anti-inflammatory IL-10 is increased, and generation of cytokine, it showed that the NOx generation decreases (see Fig. 6).

In addition, after inducing peritonitis using Zymosan A, oral administration of the lysophosphatidylethanolamine derivative according to the present invention showed that total leukocyte infiltration was effectively decreased with time (see FIG. 9 ).

Therefore, lysophosphatidylethanolamine derivatives according to the present invention inhibit the peritonitis induced by zymosan A in vivo upon oral administration, thereby producing inflammatory mediator lipids or cytokines in the abdominal cavity. It effectively inhibits and increases the production of anti-inflammatory cytokines exhibits excellent anti-inflammatory efficacy, it can be usefully used as an active ingredient in the composition for preventing and treating inflammatory diseases.

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

The administration may contain one or more active ingredients exhibiting the same or similar function. It may further comprise one or more pharmaceutically acceptable carriers for administration. The pharmaceutically acceptable carrier may be a mixture of saline, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and one or more of these components. If necessary, an antioxidant, Other conventional additives such as a bacteriostatic agent may be added. In addition, since the lysophosphatidylethanolamine derivative according to the present invention is water-soluble, it is easy to formulate a variety of formulations, for example, for injection such as an aqueous solution, suspension, emulsion, etc. by additionally adding a diluent, a dispersant, a surfactant, a binder, and a lubricant. It may be formulated into a formulation, powder, tablet, capsule, pill, granule or injection solution. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18th, 1990) in a suitable manner in the art.

The administration may be parenteral (eg, applied intravenously, subcutaneously, intraperitoneally or topically) or orally depending on the method of administration, and the dosage may be weight, age, sex, health condition, diet, administration of the patient. The range varies depending on the time, the method of administration, the rate of excretion and the severity of the disease. The daily dosage of the compound of the present invention is 0.0001 to 100 mg / kg, preferably, the amount of 0.001 to 10 mg / kg may be administered once to several times daily.

The present invention also provides an anti-inflammatory skin external preparation containing lysophosphatidylethanolamine derivative as an active ingredient.

In the external preparation for skin, the lysophosphatidylethanolamine derivative of the present invention is preferably formulated in an amount of 0.001 to 5% by weight based on the total weight of the external preparation, but is not limited thereto. Can be used together.

When the compound of the present invention is used as an external preparation for skin, it is additionally used as a fatty substance, organic solvent, solubilizer, thickening and gelling agent, emollient, antioxidant, suspending agent, stabilizer, foaming agent, fragrance, surfactant, Commonly used in water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or external preparations for the skin It may contain adjuvants conventionally used in the field of dermatology, such as any other ingredients used. In addition, the components can be introduced in amounts commonly used in the dermatology field.

In addition, the present invention provides an anti-inflammatory cosmetic composition containing a lysophosphatidylethanolamine derivative as an active ingredient.

In the composition of the present invention, the lysophosphatidylethanolamine derivative is preferably contained in an amount of 0.005 to 10% by weight, based on the total weight of the composition. Can be used.

The composition of the present invention may further contain at least one active ingredient exhibiting the same or similar function in addition to the lysophosphatidylethanolamine derivative.

The cosmetic composition of the present invention includes, but is not limited to, a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids and seaweed extracts.

The water-soluble vitamin may be any compound as long as it can be blended into cosmetics, preferably vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, vitamin H and the like. And salts thereof (thiamine hydrochloride, sodium ascorbate salt, etc.) and derivatives (ascorbic acid-2-sodium phosphate salt, ascorbic acid-2-magnesium phosphate salt and the like).

The oil-soluble vitamin may be any compound that can be formulated in cosmetics, preferably vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (DL-alpha tocopherol, D-alpha tocopherol, L-alpha tocopherol), and Derivatives (ascorbic palmitate, ascorbic stearate, ascorbicate dipalmitate, DL-alpha tocopherol acetate, DL-alpha tocopherol nicotinic acid, DL-pantothenyl alcohol, D-pantothenyl alcohol, pantothenylethyl ether, etc.) Include.

The polymer peptide may be any compound as long as it can be incorporated into cosmetics, and collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin or keratin is preferable.

The polymer polysaccharide may be any compound as long as it can be blended into cosmetics, and hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or a salt thereof (such as sodium salt) is preferable.

Any of the sphingolipids can be used as long as they can be incorporated into cosmetics, and ceramide, phytosphingosine, and sphingosaccharide lipids are preferable.

The seaweed extract may be any compound as long as it can be blended into cosmetics, and brown algae extract, red algae extract or green algae extract is preferable, and calginane, arginic acid, sodium arginate or potassium arginate purified from these seaweed extracts is preferable. Do.

The cosmetic of the present invention may be blended with the above essential components, if necessary, with other ingredients normally blended into the cosmetic. The other components include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavorings, blood circulation accelerators, Cooling agents, limiting agents or purified water are preferred, but not limited thereto.

The oil or fat component is preferably an ester oil, a hydrocarbon oil, a silicone oil, a fluorine oil, an animal oil or a plant oil. Examples of ester-based fats include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isostearyl isostearate, Butyl isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isosilyl myristate, isostearic acid isostearyl, isostearyl palmitate, octyldodecyl myristate, Trimethylol propane, triisostearic acid trimethylol propane, tetra 2-ethylhexanoic acid pentaerythritol tetra (2-ethylhexanoate) , Decyl caprylate, decyl laurate, hexyl laurate, myristate decyl, myristyl myristate, myristine monoethyl stearate, stearyl stearate, decyl oleate, ricinoleic acid tri , Isostearyl stearate, isostearyl stearate, isodecyl stearate, octyldodecyl oleate, octyldodecyl linoleate, isopropyl isostearate, isopropyl stearate, isopropyl stearate, isopropyl stearate, -Hexyl stearate, stearyl ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di (capryl, capric acid) propylene glycol, Propyleneglycol propionate, propyleneglycol propionate, dicaproic acid neopentyl glycol, dioctanoic acid neopentyl glycol, tricarboxylic acid glyceryl, triunsaturated glyceryl, triisopalmitic acid glyceryl, triisostearic acid glyceryl, neopentanoic acid octyldodecyl Octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, Sothete octylate, polyglycerol oleate, polyglycerine isostearate, triisocetyl citrate, triisoalkyl citrate, triisoctyl citrate, lauryl lactate, myritic lactate, octyl lactate, octyl lactate, tricitric acid Ethyl, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malic acid, 2-ethylhexyl hydroxystearate, di2-ethylhexyl succinate, diisobutyl adipicate, diisopropyl sebacinate, Dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, physteryl isostearate, phytic oleate, 12-Steloylhydroxystearate isocetyl, 12-steloylhydroxystearate stearyl or 12-ste Preferred esters such as alloylhydroxystearic acid isostearyl are not limited thereto.

The hydrocarbon-based oils and fats are preferably hydrocarbon-based oils such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax or waselin, but are not limited thereto.

Examples of the silicone-based oils and fats include polymethylsilicone, methylphenylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane methylcetyloxysiloxane copolymer, dimethylsiloxane methylsteoxysiloxane copolymer, and alkyl modification. Silicone oil or amino modified silicone oil is preferred but not limited thereto.

As the fluorine-based fat or oil, perfluoropolyether is preferable, but is not limited thereto.

Examples of the animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, soybean oil, soybean oil, corn oil, rapeseed oil, almond oil, palm kernel oil, palm oil, castor oil, sunflower oil, grapes. Seed oil, Cottonseed oil, Palm oil, Cuckoo nut oil, Wheat germ oil, Rice germ oil, Shea butter, Walnut colostrum, Marker demia nut oil, Meadow home oil, Egg yolk oil, Uji, Horse oil, Mink oil, Orange rape oil, Jojoba Animal or vegetable fats and oils, such as oil, candler wax, carnava wax, liquid lanolin, or hardened castor oil, are preferred, but are not limited thereto.

The moisturizing agent is preferably a water-soluble low molecular moisturizer, a fat-soluble molecular moisturizer, a water-soluble polymer or a fat-soluble polymer, but is not limited thereto. As the water-soluble low molecular moisturizer, serine, glutamine, sorbitol, mannitol, pyrrolidone sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (polymerization degree n = 2 or more), polypropylene Preferred are, but not limited to, glycol (polymerization degree n = 2 or more), polyglycerol B (polymerization degree n = 2 or more), lactic acid or lactic acid salt. The fat-soluble low molecular moisturizer is preferably cholesterol or cholesterol ester, but is not limited thereto. The water-soluble polymer is preferably a carboxyvinyl polymer, polyaspartic acid, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan or dextrin But is not limited thereto. The fat-soluble polymer is preferably, but not limited to, polyvinylpyrrolidone-eicocene copolymer, polyvinylpyrrolidone hexadecene copolymer, nitrocellulose, dextrin fatty acid ester or polymer silicone. As the emollient, long-chain acyl glutamic acid cholesteryl ester, hydroxy stearic acid cholesterol, 12-hydroxystearic acid, stearic acid, rosin acid or lanolin fatty acid cholesteryl ester are preferred, but not limited thereto.

As the surfactant, nonionic surfactants, anionic surfactants, cationic surfactants or amphoteric surfactants are preferred, but not limited thereto. Nonionic surfactants include self-emulsifying glycerin monostearate, propylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerol fatty acid esters, sorbitan fatty acid esters, POE (polyoxyethylene) sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE Glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hardened castor oil, POE castor oil, POEPOP (polyoxyethylene polyoxypropylene) copolymer, POEPOP alkyl ether, polyether modified silicone, lauric acid alkanolamide, alkyl Amine oxides or hydrogenated soybean phospholipids are preferred but not limited thereto. Anionic surfactants include fatty acid soaps, alpha-acyl sulfonates, alkyl sulfonates, alkyl allyl sulfonates, alkyl naphthalene sulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkylphosphates, and alkylamides. Phosphates, alkyloylalkyltaurine salts, N-acylamino acid salts, POE alkyl ether carboxylate salts, alkylsulfosuccinate salts, sodium alkylsulfoacetates, acylated hydrolyzed collagen peptide salts or perfluoroalkyl phosphate esters are preferred. It is not limited. As cationic surfactant, alkyl trimethylammonium chloride, stearyl trimethyl ammonium chloride, stearyl trimethyl ammonium chloride, cetostearyl trimethyl ammonium chloride, distearyl dimethyl ammonium chloride, stearyl dimethyl benzyl ammonium bromide, behenyl trimethyl ammonium chloride, chloride Benzalkonium, diethylaminoethylamide stearate, dimethylaminopropylamide stearate or lanolin derivatives Quaternary ammonium salts are preferred, but not limited thereto. As the amphoteric surfactant, such as carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amide sulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type or amideamine type, etc. Amphoteric surfactants are preferred but not limited thereto.

Examples of the organic and inorganic pigments include silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengal, clay, bentonite, titanium film mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, and oxide Inorganic pigments such as aluminum, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine, chromium oxide, chromium hydroxide, coloramine and composites thereof; Polyamide, Polyester, Polypropylene, Polystyrene, Polyurethane, Vinyl Resin, Urea Resin, Phenol Resin, Fluoro Resin, Silicon Resin, Acrylic Resin, Melamine Resin, Epoxy Resin, Polycarbonate Resin, Divinylbenzene Styrene Copolymer, Silk Organic pigments such as powder, cellulose, CI pigment yellow, and CI pigment orange, and composite pigments of inorganic pigments and organic pigments thereof are preferable, but are not limited thereto.

As said organic powder, Metal soaps, such as a calcium stearate; Metal salts of alkyl phosphates such as sodium zinc cetylate, zinc laurylate and calcium lauryl laurate; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc and N-lauroylglycine calcium; Amidosulfonic acid multivalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; Such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylidene, N-alpha-paratyylnitin, N-alpha-lauroyl arginine, N- Acyl basic amino acids; N-acylpolypeptides such as N-lauroylglycylglycine; Alpha-amino fatty acids such as alpha-aminocaprylic acid, alpha-aminoaurauric acid, and the like; Or polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene styrene copolymer, ethylene tetrafluoride, but is not limited thereto.

As said ultraviolet absorber, paraamino benzoic acid, ethyl paraamino benzoate, amyl paraamino benzoate, octyl paraamino benzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylic acid, and cinnamic acid Benzyl, paramethoxy cinnamic acid-2-ethoxyethyl, paramethoxy cinnamic acid octyl, diparamethoxy cinnamic acid mono-2-ethylhexaneglyceryl, paramethoxy cinnamic acid isopropyl, diisopropyl diisopropyl cinnamic acid ester mixture, wuro Canonic acid, ethyl urokanoate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and salts thereof, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenone sodium sulfonate, dihydroxybenzophenone , Tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p- (carbo-2'-ethylhexyl-1'-oxy) -1 , 3,5-triazine or 2- ( 2-hydroxy-5-methylphenyl) benzotriazole is preferred, but not limited thereto.

Examples of the fungicides include hinokithiol, trichloric acid, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcinol, isopropylmethylphenol, azulene, salicylic acid, zinphylthione, benzalkonium chloride, Photosensitizer No. 301, mononitroguicol sodium or undecylenic acid are preferred, but not limited thereto.

Preferred antioxidants include, but are not limited to, butylhydroxyanisole, propyl gallic acid, or elisorbic acid.

The pH adjusting agent is preferably, but is not limited to, citric acid, sodium citrate, malic acid, sodium malate, fmaric acid, sodium fmarate, succinic acid, sodium succinate, sodium hydroxide or sodium hydrogen phosphate.

The alcohol is preferably a higher alcohol such as cetyl alcohol, but is not limited thereto.

In addition, the compounding component which may be added other than this is not limited to this, Moreover, Although all said components can be mix | blended within the range which does not impair the objective and effect of this invention, it is mix | blended in 0.01 to 5% weight percentage with respect to gross weight. Preferably, it is more preferably blended at 0.01 to 3% by weight, but is not limited thereto.

The cosmetic of the present invention may take the form of a solution, an emulsion or a viscous mixture, but is not limited thereto. The components included in the cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the compound of the present invention as an active ingredient, and common auxiliaries and carriers such as stabilizers, solubilizers, vitamins, pigments and flavorings. It may include, but is not limited to.

The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, but may be prepared in an emulsion, cream, lotion, pack, foundation, lotion, essence or hair cosmetic, but is not limited thereto. Specifically, the cosmetic composition of the present invention skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisturizing cream, hand cream, foundation, essence, nutrition essence, Formulations of packs, soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions or body cleansers.

When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, waxes, paraffins, starches, tracants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. However, it is not limited thereto.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant, such as butane or dimethyl ether.

In the case of the solution or emulsion of the present invention, a solvent, a solvating agent or an emulsifying agent is used as a carrier component, and water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, Fatty acid esters of 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan may be used, but are not limited thereto.

When the dosage form of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant may be used but is not limited thereto.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide But are not limited to, ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are illustrative of the present invention, and the contents of the present invention are not limited by the following examples.

< Example  1> 2- ARA - lysoPtdEtn Manufacturing

C18 (plasm) -20: 4 PtEtn (1- (1Z-octadecenyl) -2-arachidonoyl-sn-glycero-3-phosphoethanolamine) was added 2.5 M HCl (0.8 ml), methanol (2 ml) and chloroform (1 ml). It was added to a mixed solvent of and suspended by stirring at room temperature for 20 minutes. The hydrolysis product from the reaction mixture was extracted by Bligh and Dyer method and then separated and purified by silica gel TLC (elution solvent-chloroform: methanol: water = 65: 25: 4). Eventually the band containing lysoPtdEtn was eluted, which was extracted with methanol and stored at -80 ° C until use.

< Example  2> 2- DHA - lysoPtdEtn Manufacturing

Example 1 except for using C18 (plasm) -20: 4 PtEtn instead of C18 (plasm) -22: 6 PtEtn (1- (1Z-octadecenyl) -2-docosahexaenoyl-sn-glycero-3-phosphoethanolamine) It was carried out in the same way as.

< Example  3> 2- (15- HETE ) - lysoPtdEtn Manufacturing

2-ARA-lysoPtdEtn (200 μM) prepared in Example 1 was added to 5 ml of borax buffer (50 mM, pH 9.0), soybean LOX-1B (4 KU / ml) After oxidizing for 1 hour, 1 mM SnCl _{2 was added} to the peroxide, followed by stirring at room temperature for 10 minutes to reduce the hydroxide. The hydroxide product was extracted by Bligh and Dyer method and then reversed-HPLC (elution solvent-acetonitrile: water: formic acid = 60:40) using Zorbrax eclipse XDB C18 column (5 μm, 50 × 4.6 mm, Agilent Technologies, USA) : 0.1) was carried out to separate 2- (15-HETE) -lysoPtdEtn, and the amount of separated 12- (15-HETE) -lysoPtdEtn was calculated from an extinction coefficient at 234 nm (E1 m, 1 cm = 28,000). The 15-hydroxyaicosapentaenoyl lysophosphatidylcholine was stored at −80 ° C. before use.

< Example  4> 2- (17- HDHE ) - lysoPtdEtn Manufacturing

Except for using 2-DHA-lysoPtdEtn prepared in Example 2 instead of 2-ARA-lysoPtdEtn was carried out in the same manner as in Example 3.

LC Of ESI - MS  Structure identification

LC-ESI-MS analysis was performed to confirm the formation of peroxides by the 15-LOX catalyst of 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn prepared in Examples 1 or 2. The LC-ESI-MS analysis was performed using an MSDI spectrometer (HP 1100 series LC / MSA, Hewlett Packard, USA) mounted on a Zorbrax eclipse XDB C18 column (5 μm, 50 × 4.6 mm, Agilent Technologies, USA) It was.

Full scan mass spectrometry at 234 nm was performed in the m / z 250-700 range and data acquisition was performed in positive mode and negative mode ().

The measurement results are shown in Fig .

1A shows a peak with a retention time of 1.6 minutes, whereby the mass spectrum of the compound corresponding to the peroxide derivative of 2-ARA-lysoPtdEtn is shown. In FIG. 1B , 532 (MH) ⁻ in negative mode and 500 (in positive mode) are shown . Mass to charge ratios of M + HH ₂ OO) ⁺ , 516 (M + HH ₂ O) ⁺ , 534 (M + H) ⁺ , 556 (M + Na) ⁺ , and 572 (M + K) ⁺ were shown.

Likewise, in FIGS. 1C and 1D , the mass spectrum of the compound corresponding to the peroxide derivative of 2-DHA-lysoPtdEtn was shown.

From this, it can be seen that 2-ARA-lysoPtdEtn and 2-DHA-lysoPtdEtn are converted to their peroxides by soy LOX catalyst, and through this process 2- (15-HETE) -lysoPtdEtn and 2- (17-HDHE)- You can see that lysoPtdEtn is created.

< Experimental Example  1> of the present invention Lysophosphatidylethanolamine  Derivative Zymosan  Inhibitory Effect of A (zymosan A) Induced Plasma Outflow

5-Lipoxygenase (5-LOX) plays an important role in the production of leukotriene, an inflammatory mediator from arachidonic acid, and thus leads to a decrease in inflammatory response through inhibition of 5-LOX. Has been reported. In addition, zymosan induced peritonitis is known to be mediated by leukotriene produced by 5-LOX (RaoTS, et al., J Pharmacol Exp Ther. 1994 Jun; 269 (3)). : 917-25; Doherty NS, et al., Prostaglandins. 1985 Nov; 30 (5): 769-89).

Accordingly, the present inventors, in order to determine the inhibitory effect of lysophosphatidylethanolamine derivatives according to the present invention on the zymoic acid-induced peritonitis, Zymosan A (Saccharomyces cerevisiae) (Sigma-Aldrich Corp, St. Louis, MO, USA) ) Induces intraperitoneal inflammation, and then the anti-inflammatory effect was evaluated by oral administration of each lysophosphatidylethanolamine derivative.

Specifically, mice reared under SPF conditions (6 weeks old, males) were kept in 12 hours light-cycle cycle conditions in animal storage rooms at the College of Pharmacy 12 hours after delivery, followed by an intraperitoneal inflammatory response experiment. First, in each group of mice consisting of 5 to 10 mice, 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn prepared in Example 1 or 2 as test groups was diluted in cold PBS at a dose of 50 or 150 μg / kg. Oral administration. One hour later, Zymosan A (1 mg / mouse) dissolved in saline was administered intraperitoneally, followed by injection of 5% Evans Blue dye solution (100 μl / mouse) into the tail vein.

Two hours after administration, the mice were cervically cut under mild anesthesia [ether or fluorane], 4 ml of ice-cold PBS was intraperitoneally injected, and the peritoneal fluid was recovered by a plastic syringe and centrifuged. (3,000 rpm, 10 minutes). The anti-inflammatory effect of the supernatant obtained by centrifugation was measured by measuring the exudation of Evans blue at 610 nm using a spectrophotometer (Arita M, et al., Proc Natl Acad). Sci US A. 2005 May 24; 102 (21): 7671-6; Takenaka M, et al., Biol Pharm Bull. 2005 Jul; 28 (7): 1291-3).

The results are shown in Figure 2 and Table 1.

Plasma outflow inhibition rate (%) ED ₅₀ (μg / kg) 50 μg / kg 150 μg / kg Example 1 17 33 221 Example 2 To 42 ~ 58 111

As shown in FIG . 2 and Table 1, 2-ARA-lysoPtdEtn and 2-DHA-lysoPtdEtn reduced concentration of Zymosan-induced plasma efflux. Therefore, the lysophosphatidylethanolamine derivative of the present invention has an inhibitory effect on the plasma outflow induced by Zymosan A and confirmed the anti-inflammatory effect.

< Experimental Example  2> of the present invention Lysophosphatidylethanolamine  Derivative Zymosan  Inhibition of A-induced Leukocyte Outflow Inhibition

Leukocyte leakage is a phenomenon that occurs continuously after reversible opening of the endothelial cell dense joint at the onset of acute inflammation (Serhan CN et al., J Exp Med (2002) 196: 1025-1037). Therefore, in order to determine the ability of lysophosphatidylethanolamine derivatives according to the present invention to control the zymosan A-induced inflammation, the effect of inhibiting zymosan A-induced leukocyte leakage was measured as follows.

Specifically, in each group of mice, 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn was diluted in cold PBS at a dose of 50 or 150 μg / kg as a test group, and then diluted in PBS with PD146176 (15-lipoxygenas inhibitor) or PD146176. It was administered orally without. Then, after 1 hour, Zymoic acid A (1 mg / mouse) dissolved in saline was administered intraperitoneally. After 4 hours, the peritoneal lavage was then stained using tryphane blue dye, and the total number of white blood cells was measured using a light microscope. The results were expressed as mean ± SEM (n = 5), expressed as # for p <0.001 compared to PBS treated group, and * for p <0.05 compared to PBS treated group, and p <0.01 compared to PBS treated group. The case was marked ** and the case marked with significant differences between treatment groups. The results are shown in FIG. 3 and Table 2.

Leukocyte leakage inhibition rate (%) ED ₅₀ (μg / kg) 50 μg / kg 150 μg / kg Example 1 34 33 221 Example 2 42 ~ 58 111

As shown in FIG . 3 and Table 2, orally administered 2-ARA-lysoPtdEtn and 2-DHA-lysoPtdEtn reduced concentrations of zymoacid-induced leukocyte leakage and administered Zymosan A at 150 μg / kg The lysophosphatidylethanolamine derivatives of the present invention exhibited an inhibitory effect on leukocyte efflux induced by Zymosan A, thereby confirming the anti-inflammatory effect. .

< Experimental Example  3> Lysophosphatidylethanolamine  Bio of Derivatives Abdominal cavity  Identification of effects on the production of inflammatory lipid mediators

In order to confirm the effect of lysophosphatidylethanolamine derivatives according to the present invention on the production of inflammatory lipid mediators in vivo in vivo, the contents of mouse intraperitoneal inflammatory lipid mediators LTC ₄ and LTB ₄ by Zymosan A induction were measured.

Specifically, mice in each group were administered orally by diluting 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn to 50 or 150 μg / kg dose in cold PBS. Then, after 1 hour, Zymoic acid A (1 mg / mouse) dissolved in saline was administered intraperitoneally. After 4 hours, peritoneal lavage was performed and the contents of proinflammatory lipid mediators LTC ₄ and LTB ₄ in the cell-free supernatant were measured using an enzyme immunoassay kit (EIA). Each sample was measured twice to determine the measured value. The results were expressed as mean ± SEM (n = 5), expressed as # for p <0.001 compared to PBS treated group, and * for p <0.05 compared to PBS treated group, and p <0.01 compared to PBS treated group. In the case indicated by **.

The results are shown in FIG. 4 and Table 3.

EC ₅₀ (μg / kg) LTC ₄ LTB ₄ Example 1 132 139 Example 2 84 211

As shown in FIG . 4 and Table 3, orally administered 2-ARA-lysoPtdEtn and 2-DHA-lysoPtdEtn are concentration dependent, producing leukotriene C ₄ (LTC ₄ ) and leukotriene B ₄ (LTB ₄ ) It can be seen that it inhibits the production of.

Therefore, the lysophosphatidylethanolamine derivative of the present invention exhibits an effect of significantly inhibiting the production of intraperitoneal inflammatory lipid mediators induced by Zymosan A, thereby confirming the anti-inflammatory effect.

< Experimental Example  4> Lysophosphatidylethanolamine  Derivative Zymosan  A induction Abdominal cavity  Inflammation Cytokines  Check the suppression effect on generation

To confirm the effect of lysophosphatidylethanolamine derivatives according to the present invention on the production of in vivo intraperitoneal inflammatory cytokines, cytokines IL-1, IL-6 and TNF-, which are intraperitoneal inflammation mediators of mice induced by Zymosan A The content of α and the content of IL-10 as anti-inflammatory cytokines were measured.

Specifically, mice in each group were administered orally by diluting 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn to 50 or 150 μg / kg dose in cold PBS. Then, after 1 hour, Zymoic acid A (1 mg / mouse) dissolved in saline was administered intraperitoneally. After 4 hours, the peritoneal lavage was performed and the contents of the respective cytokines IL-1, IL-6, TNF-α, and IL-10 were measured using an enzyme immunoassay kit (EIA). Each sample was measured twice to determine the measured value. The results were expressed as mean ± SEM (n = 5), expressed as # for p <0.001 compared to PBS treated group, and * for p <0.05 compared to PBS treated group, and p <0.01 compared to PBS treated group. In the case indicated by **.

The results are shown in Fig.

As shown in FIG . 5 , the results of the measurement showed that the oral administration of lysophosphatidylethanolamine derivatives according to the present invention significantly induces the production of in vivo intraperitoneal inflammatory cytokines IL-1, IL-6 and TNF-α according to the dosage. The anti-inflammatory cytokine IL-10 was found to increase significantly with dose.

Therefore, it can be seen that the lysophosphatidylethanolamine derivative according to the present invention exhibits an anti-inflammatory effect by inhibiting cytokines causing inflammation and increasing anti-inflammatory cytokines.

< Experimental Example  5> Lysophosphatidylethanolamine  Derivative Zymosan  Identification of inhibitory effects on A-induced nitric oxide production

Nitric oxide (NO) plays an important role in inflammatory processes, particularly including the regulation of inflammatory cells inside the site of inflammation and the upregulation of vascular permeability. In Zymosan A induced peritonitis, nitric oxide is produced by activated macrophages and eventually oxidized to nitrite or nitrate within seconds (Guzik TJ et al., J Physiol Pharmacol (2003) 54: 469-487). Therefore, in order to examine the anti-inflammatory effect of the lysophosphatidylethanolamine derivative of the present invention, the production of Zymosan A induced nitric oxide was measured as follows.

Specifically, 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn prepared in Examples 1 or 2 as test groups were diluted orally administered to cold PBS at a dose of 50 or 150 μg / kg. Then, after 1 hour, Zymoic acid A (1 mg / mouse) dissolved in saline was administered intraperitoneally. After 4 hours, the peritoneal lavage was performed and the content of nitric oxide in the cell-free supernatant was measured using colorimetric method. The results were expressed as mean ± SEM (n = 5), expressed as # for p <0.001 compared to PBS treated group, and * for p <0.05 compared to PBS treated group, and p <0.01 compared to PBS treated group. The case is marked **.

The results are shown in FIG. 6 and Table 4.

Nitric oxide inhibition rate (%) 50 μg / kg 150 μg / kg Example 1 60 70 Example 2 89 79

As shown in FIG . 6 and Table 4, orally administered 2-ARA-lysoPtdEtn and 2-DHA-lysoPtdEtn significantly inhibited nitric acid-induced nitric oxide production. Therefore, the lysophosphatidylethanolamine derivative of the present invention exhibits an inhibitory effect on the production of nitric oxide produced in inflammation, thereby confirming the anti-inflammatory effect.

< Experimental Example  6> Lysophosphatidylethanolamine  Derivative Zymosan  Confirmation of A-induced Plasma Outflow Inhibitory Effect

Experimental Examples 1 and 2 except that the dosage was 15, 50, 150 μg / kg with 2- (15-HETE) -lysoPtdEtn and 2- (17-HDHE) -lysoPtdEtn prepared in Example 3 or 4. Zymosan A-induced plasma efflux inhibition experiments were performed in the same manner and the results are shown in FIGS. 7 and 5.

Plasma outflow inhibition rate (%) ED ₅₀ (μg / kg) 15 μg / kg 50 μg / kg 150 μg / kg Example 3 77 49 38 48 Example 4 62 47 35 42

As shown in FIG . 7 and Table 5, 2- (15-HETE) -lysoPtdEtn and 2- (17-HDHE) -lysoPtdEtn effectively reduced plasma efflux induced by zymoic acid in a concentration dependent manner. Therefore, the lysophosphatidylethanolamine derivative of the present invention has an inhibitory effect on the plasma outflow induced by Zymosan A and confirmed the anti-inflammatory effect.

< Experimental Example  7> Lysophosphatidylethanolamine  Derivative Zymosan  Inhibition of A-induced Leukocyte Outflow

Experimental Example 2 with the exception that the dosage was 15, 50, 150 μg / kg with 2- (15-HETE) -lysoPtdEtn and 2- (17-HDHE) -lysoPtdEtn prepared in Example 3 or 4 Zymosan A-induced leukocyte leakage inhibition experiment was performed in the same manner, and the results are shown in FIGS. 8 and 6.

Leukocyte leakage inhibition rate (%) ED ₅₀ (μg / kg) 15 μg / kg 50 μg / kg 150 μg / kg Example 3 55 23 23 28 Example 4 46 22 15 26

As shown in FIG . 8 and Table 6, 2- (15-HETE) -lysoPtdEtn and 2- (17-HDHE) -lysoPtdEtn effectively reduced the leukocyte induced leukocyte efflux in a concentration dependent manner. Therefore, the lysophosphatidylethanolamine derivative of the present invention exhibits an inhibitory effect on leukocyte outflow induced by Zymosan A and confirmed the anti-inflammatory effect.

< Experimental Example  8> Lysophosphatidylethanolamine  Derivative Zymosan  Confirmation of A-induced Peritonitis Inhibitory Effect

In order to investigate whether the lysophosphatidylethanolamine derivative according to the present invention can attenuate Zymosan A-induced peritonitis, a Zymosan A-induced leukocyte infiltration experiment was performed in ICR-based mice as follows.

Specifically, Zymosan A (1 mg / mouse) dissolved in physiological saline was administered intraperitoneally of mice. Then, when inflammation was at its peak (12 hours after Zymosan A treatment), no treatment was performed as a control or orally administered 2-ARA-lysoPtdEtn or 2-DHA-lysoPtdEtn removed in Examples 1 or 2. 15, 20 and 24 hours after the time of administration, the samples which were intraperitoneally washed were collected and stained using tryphane blue dye, and the total number of infiltrated leukocytes was measured using a light microscope. The results were expressed as mean ± SEM (n = 5), and when the 2-ARA-lysoPtdEtn treatment group was p <0.05 compared to the Zymosan A treatment group, it was *, and 2-DHA- compared to the Zymosan A treatment group. The lysoPtdEtn treatment group was expressed as + for p <0.05 and ++ for p <0.01. The results are shown in FIG. 9 and Table 7.

Leukocyte infiltration inhibition rate (%) After 15 hours After 20 hours After 24 hours Example 1 23 21 33 Example 2 29 33 44

As shown in Figure 9 and Table 7, orally administered 2-ARA-lysoPtdEtn and 2-DHA-lysoPtdEtn were administered at the peak of inflammation, effectively reducing the total leukocyte infiltration number. Therefore, the lysophosphatidylethanolamine derivative of the present invention can be usefully used as an anti-inflammatory composition by effectively inhibiting Zymosan A induced peritonitis.

< Manufacturing example  1> Preparation of Pharmaceutical Formulations

<1-1> Sanje  Produce

Lysophosphatidylethanolamine derivative 0.1 mg

Lactose 100 mg

After mixing the above components, the mixture was packed in an airtight container to prepare a powder.

<1-2> Preparation of tablets

Lysophosphatidylethanolamine derivative 0.1 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

&Lt; 1-3 > Preparation of capsules

Lysophosphatidylethanolamine derivative 0.1 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

<1-4> Production of liquid agent

Lysophosphatidylethanolamine derivative 0.2 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

<1-5> Preparation of Injection

Lysophosphatidylethanolamine derivative 100 μg / ml

    Dilute hydrochloric acid BP until pH 7.6

    Main sodium chloride BP up to 1 ml

The lysophosphatidylethanolamine derivative was dissolved in an appropriate volume of main sodium chloride BP, the pH of the resulting solution was adjusted to pH 7.6 with dilute hydrochloric acid BP, and the volume was adjusted with a main volume of sodium chloride BP and mixed well. The solution was filled into a 5 ml Type I ampoule of clear glass, encapsulated under an upper grid of air by dissolving the glass, and autoclaved at 120 ° C. for at least 15 minutes to prepare an injection.

< Manufacturing example  2> Manufacturing of food

Food containing the lysophosphatidylethanolamine derivative of the present invention was prepared as follows.

<2-1> Production of flour food

0.01 to 0.1 parts by weight of the lysophosphatidylethanolamine derivative of the present invention was added to flour, and bread, cake, cookies, crackers and noodles were prepared using this mixture to prepare foods for health promotion.

<2-2> Dairy products ( dairy products Manufacturing

0.01 to 0.1 parts by weight of the lysophosphatidylethanolamine derivative of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<2-3> Preparation of Wire

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh. Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer. The lysophosphatidylethanolamine derivative of the present invention was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by spraying and drying with a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder. The dry grains of the grains, seeds and 15-hydroxyaicosapentaenoyl lysophosphatidylcholine prepared above were prepared by combining the following ratios.

(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)

Lysophosphatidylethanolamine derivative (0.1 parts by weight),

(0.5 parts by weight), and

Rhubarb (0.5 parts by weight).

<2-4> Production of health supplement

Lysophosphatidylethanolamine derivative 1 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

Biotin 10 μg

Nicotinamide 1.7 mg

Folic acid 50 μg

Calcium Pantothenate 0.5 mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

15 mg potassium monophosphate

Dicalcium Phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

< Manufacturing example  3> Health drink  Produce

    Lysophosphatidylethanolamine derivative 1 mg

    Citric acid 100 mg

    Oligosaccharide 100 mg

    Plum concentrate 2 mg

    Taurine 100 mg

    Add 500 ml of purified water

After mixing the above components in accordance with a conventional healthy beverage production method, and then stirred and heated at 85 for about 1 hour, the resulting solution is filtered and obtained in a sterilized 1 L container, sealed sterilization and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions. Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

Preparation Example 4 Preparation of Cosmetics

<4-1> Cosmetic Preparation of Emulsion Formulation

Cosmetics of the emulsifier type were prepared with the composition shown in Table 1. The production method is as follows.

1) The mixture which mixed the raw materials of 1-9 was heated at 65-70 degreeC.

2) 10 was added to the mixture of step 1).

3) The mixture of the raw materials of 11-13 was heated at 65-70 degreeC, and was completely dissolved.

4) While passing through step 3), the mixture of 2) was slowly added to emulsify for 2 to 3 minutes at 8,000 rpm.

5) The raw material of 14 was dissolved in a small amount of water, and then added to the mixture of step 4) and emulsified for 2 minutes.

6) 15 to 17 of the raw materials were weighed, respectively, and added to the mixture of step 5) and emulsified for 30 seconds.

7) After emulsifying the mixture of step 6) through a degassing process to prepare a cosmetic of emulsion type by cooling to 25 ~ 35 ℃.

Composition of Emulsifying Formulations 1, 2, 3 Furtherance Emulsifier 1 Emulsifier 2 Emulsifier 3 One Stearic acid 0.3 0.3 0.3 2 Stealy alcohol 0.2 0.2 0.2 3 Glyceryl monostearate 1.2 1.2 1.2 4 Wax 0.4 0.4 0.4 5 Polyoxyethylene sorbitan monolauric acid ester 2.2 2.2 2.2 6 Methyl paraoxybenzoate 0.1 0.1 0.1 7 P-hydroxybenzoic acid profile 0.05 0.05 0.05 8 Cetyl ethyl hexanoate 5 5 5 9 Triglyceride 2 2 2 10 Cyclomethicone 3 3 3 11 Distilled water ~ 100 ~ 100 ~ 100 12 Concentrated glycerin 5 5 5 13 Triethanolamine 0.15 0.15 0.15 14 Polyacrylic acid polymer 0.12 0.12 0.12 15 Pigment 0.0001 0.0001 0.0001 16 incense 0.10 0.10 0.10 17 Lysophosphatidylethanolamine derivative 0.001 0.01 0.1

<4-2> Cosmetic Preparation of Solubilized Formulation

A cosmetic of solubilized formulation was prepared with the composition shown in Table 2. The production method is as follows.

1) Raw materials 2 to 6 were placed in raw material 1 (purified water) and dissolved using a still mixer.

2) The raw materials of 8 to 11 were put into the raw material of 7 (alcohol) and completely dissolved.

3) The mixture of step 2) was solubilized with slow addition to the mixture of step 1).

Solubilization  Composition of Formulations 1, 2, 3 Furtherance  Solubilized Formulation 1 Solubilized Formulation 2  Solubilized Formulation 3 One Purified water  ~ 100  ~ 100  ~ 100 2 Concentrated glycerin  3  3  3 3 1,3-butylene glycol  2  2  2 4 EDTA-2Na  0.01  0.01  0.01 5 Pigment  0.0001  0.0002  0.0002 6 Lysophosphatidylethanolamine derivative  0.1 0.2 0.5 7 Alcohol (95%)  8 8 8 8 Methyl paraoxybenzoate  0.1  0.1  0.1 9 Polyoxyethylene
Hydrogenated Ester  0.3  0.3  0.3 10 incense  0.15  0.15  0.15 11 Cyclomethicone  -  -  0.2

Claims (8)

Risophosphatidylethanolamine derivative represented by any one of the following Chemical Formulas 3 to 4 or a pharmaceutically acceptable salt thereof:
(3)

[Chemical Formula 4]

.
A pharmaceutical composition for the prevention and treatment of inflammatory diseases, comprising the lysophosphatidylethanolamine derivative of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
The composition for preventing and treating inflammatory diseases according to claim 2, wherein the inflammatory disease is any one selected from the group consisting of asthma, periodontitis, stomatitis, peritonitis, gastritis, enteritis, arthritis, nephritis, hepatitis and degenerative diseases. .
The composition for preventing and treating inflammatory diseases according to claim 2, wherein the composition is used as an oral preparation or injection.
Claim 1 lysophosphatidyl ethanolamine derivative or a pharmaceutically acceptable salt thereof as an active ingredient for preventing and improving inflammatory diseases health food.
The method of claim 5, wherein the inflammatory disease is any one selected from the group consisting of asthma, periodontitis, stomatitis, peritonitis, gastritis, enteritis, arthritis, nephritis, hepatitis and degenerative diseases food.
An anti-inflammatory skin external preparation containing the lysophosphatidylethanolamine derivative of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
An anti-inflammatory cosmetic composition comprising the lysophosphatidylethanolamine derivative of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

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