KR101491493B1 - Anti-inflammatory pharmaceutical composition comprising tangerine pericarp and Psidium guajava extract - Google Patents

Abstract

The present invention relates to a composition for the treatment or prevention of inflammation, which comprises a citrus peel extract (preferably parchment crust extract) and a guava leaf extract together as an active ingredient. The present invention also provides a mixing weight ratio of the citrus peel extract (preferably parchment skin extract) and guava leaf extract exhibiting a synergistic effect in achieving an inflammatory treatment or prevention effect. The present invention is also based on the fact that the antiinflammatory effect is reduced when such extract is substantially contained in the involutional extract.

Description

The present invention relates to an anti-inflammatory pharmaceutical composition comprising tangerine pericarp and Psidium guajava extract,

FIELD OF THE INVENTION The present invention relates to a pharmaceutical composition having anti-inflammatory activity, and more particularly, to a combination composition in which a plurality of active ingredients are contained to synergistically enhance anti-inflammatory activity. The present invention also relates to mixing ratios for the synergistic effects of the plurality of effective ingredients.

Inflammatory responses are a series of immune responses that necessarily occur by activated immune cells. When the immune cells are exposed to foreign substances such as microorganisms including bacteria, viruses and foreign substances in the living body, the immune cells are activated and the activated immune cells accelerate the inflammation reaction. When the inflammatory reaction by lipopolysaccharide (LPS), which is a bacterial inflammation inducer, occurs, various inflammatory factors (pro-inflammatory mediators) are produced. The inflammatory factors include inducible nitric oxide synthase (iNOS) the nitric oxide produced by the like (NO, nitric oxide) and cyclooxygenase (COX-2, cyclooxygenase) prostaglandin _{E 2 (PGE 2, prostaglandin E} 2) produced by the.

NO is a biologically active molecule with high reactivity and is produced from L-arginine by nitric oxide synthase (NOS). NO is involved in neurotransmission, vascular relaxation and cell-mediated immune responses, particularly when the macrophage is stimulated with lipopolysaccharide (LPS) and interferon-γ (IFN-γ), inducible nitric oxide synthase (iNOS) Resulting in a large amount of NO. The large amount of NO produced in this way serves as an inflammatory mediator.

PGE ₂ is a major inflammatory mediator and is involved in the inflammatory response by stimulating macrophages such as cytokines, tumor necrosis factor-α (TNF-α), and lipopolysaccharide (LPS) ), Resulting in excessive production when cyclooxygenase-2 (COX-2) is expressed. PGE ₂ may increase vascular permeability and cause heat, swelling and pain in inflamed areas. PGE ₂ is inhibited by non-steroidal anti-inflammatory drugs (NSAIDs). It is used for antipyretic, analgesic and anti-inflammatory effects. It is an anti-inflammatory agent and is used for the treatment of muscle skeletal diseases such as rheumatoid arthritis, osteoarthritis and ankylosing spondylitis. Is used.

There has been much effort to develop a pharmaceutical composition having an antiinflammatory effect by inhibiting the production of NO and PGE ₂ , and a lot of studies have been made on a pharmaceutical composition for treating or preventing inflammation including a natural plant active ingredient have.

The citrus (tangerine) is the name of a specific fault in the shared horticulture in Japan, Korea and China terminology. The content of citrus contains some vitamin C as a main ingredient though it differs somewhat depending on varieties. Vitamin A is carotene content, 60mg / 100g of early-life Wenzhou, usually about 12mg / 100g Wenzhou, and other vitamins have effects such as vitamin P and Hesperidin contains vitamin E. Citrus fruits include many physiologically active substances such as flavonoids, carotenoids, coumarins, phenylpropanoids and limonoids. Jeju native native citrus fruit ( Citrus hassaku ) is a species belonging to Citrus ( Citrus ), and has been shown to be effective for the treatment and prevention of various diseases.

The content of naringin and hesperidin in citrus peel is higher than that of pulp, and the total dietary fiber content is also higher in the perilla than in the pulp. Citrus peels are rich in carotenoids, bioflavonoids, and terpenes, which have been shown to prevent hypertension, reduce LDL cholesterol in the blood, increase HDL cholesterol content, and prevent or ameliorate circulatory diseases have. The contents of flavonoids, antioxidants, vitamins, and dietary fiber were the highest among the whole citrus fruits, peel and flesh of three citrus samples, and antioxidant activity and weight loss were also reported.

Guava is a tropical plant of the genus Psidium ( Myrtaceae ) in the United States. It is a medicinal plant that can use potted plants as well as leaves, bark and fruit for healthful and medicinal purposes. Guava contains vitamins and minerals such as vitamin C, magnesium, potassium, and calcium. It contains a number of vitamins and minerals including antioxidative mutagenesis (cancer prevention), hydrogen peroxide scavenging (anti-aging) (Amylase inhibition), anti-diabetic effect (inhibition of glucose absorption), and antiallergic action (inhibition of histamine).

Mugwort is an edible plant native to the whole country. It is a food resource rich in proteins, lipids, fiber, calcium carbonate, phosphorus, and vitamins A, B and C. From ancient times, it has been widely consumed in foods such as herbs, mugwort, porridge and mugwort cake. Among injin (茵蔯) Artemisia is a perennial plant type deciduous shrub belonging to the genus Artemisia Asteraceae, raw dangssuk, aedang mugwort, sacheolssuk (Artemisia capillaries THUNB.), Artillery, Artemisia iwayomogi ), white wormwood ( Artemisa sacrorum subsp. Kitamura), hair wormwood ( Artemisa sacrorum subsp. There is manshuria. vestita Kitamura). In particular, young leaves are called starch or heat, and they have been traditionally used in the private sector as edible, medicinal, and single medicine because of their unique fragrances and medicinal properties. It is widely used for acute hepatitis, chronic hepatitis, liver cirrhosis and liver cancer, and it is also used for diseases such as nuts and gall stones. The pharmacological action is excellence of bile secretion promoting action, hepatic function protecting action, especially hepatocyte regeneration action. Lipid degradation, coronary artery dilation and blood pressure lowering effect, fever, diuretic, antibacterial effect is also seen.

Korean Patent Registration No. 10-0642744 discloses a method of using anti-inflammatory agents in a jinjia (citrus) peel extract having physiological activity and foods using the same, and Korean Patent Publication No. 10-0847103 discloses a guava fermented composition And Korean Patent Registration No. 10-0792613 discloses a method for increasing the content of coumarin and scopollen in an antiinflammatory agent.

The present inventors have found that the anti-inflammatory effect of each of the above natural extracts has not yet reached a satisfactory level, and have conducted a steady study on a method of enhancing the anti-inflammatory effect of such natural extracts.

That is, a technical object to be achieved by the present invention is to provide a pharmaceutical composition with improved therapeutic or preventive effects of inflammation and a method for producing the same.

In order to accomplish the above object, the present invention provides a pharmaceutical composition for treating or preventing inflammation, which comprises citrus peel extract and guava leaf extract as an active ingredient. The complex active ingredients of the composition of the present invention synergistically inhibit the production of NO and PGE ₂ and consequently provide a composite composition in which the anti-inflammatory effect is enhanced more than the respective extracts.

That is, the present invention is based on the surprising fact that the composition containing citrus peel extract and Guava leaf extract at the same time exhibits a very rapid increase in the anti-inflammatory effect than when administered alone.

In the present invention are citrus Satsuma Mandarin (Citrus unshue , citrus hassaku), binary-oriented (Citrus sinensis x reticulata ), Citrus sinensis , Citrus reticulata , Citrus aurantium L, Citrus unshiu S. Marcov., Citrus aurantium L., Fortunella margarita ) and the like to those of ordinary skill in the art to which the present invention belongs, Citrus It refers to the fruit of the plant. Preferably, Citrus hassaku , a native citrus fruit of Jeju Island, is preferred as the citrus fruit of the present invention for the purpose of anti-inflammation of the present invention.

More specifically, the citrus fruit of the present invention is not limited to citrus fruits produced in a part of Jeju Island centered on Seogwipo, but also citrus fruits harvested in Jeju Island below 200 m in altitude and in Tongyoung, Goheung, Wando, Geoje, Can be used. The citrus peel of the present invention can be obtained from the by-product of citrus fruit which occupies 20 to 25% of the total citrus distributed in Korea.

Guava (Psidium according to the invention guajava ) leaves can be obtained from the agricultural by-products that occur in horticultural agriculture.

"Extracts" according to the present invention may be prepared using extraction methods well known to those of ordinary skill in the art. That is, citrus peel or guava leaves are shredded and shredded, and the extracting solvent is extracted by adding 1 to 20 times the volume of citrus peel or guava leaf pulverized product, and then extracted (concentrated), dried or purified can do.

Examples of the extraction solvent include purified water; Lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propyl alcohol and butyl alcohol; Polyhydric alcohols such as glycerin, butylene glycol and propylene glycol; Hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether and dichloromethane; Or a mixed solvent thereof may be used.

More specifically, an extraction solvent is added to extract an active ingredient substance as in the above-mentioned method, (a) heating at a temperature of 50 to 100 ° C for 1 to 20 hours in a state where a cooling condenser is installed to prevent evaporation of the solvent, (B) immersing the mixture at a temperature of 5 to 37 DEG C for 1 to 15 days to extract the active ingredient. That is, it can be produced by an extraction method such as stirring extraction, reflux cooling extraction, cold extraction, ultrasonic extraction or supercritical extraction.

Preferably, the present invention relates to an anti-inflammatory composition comprising a citrus peel extract and a guava leaf extract, wherein the citrus peel extract and the guava leaf extract are mixed at a weight ratio of 87: 13 to 62:38, more preferably 80:20 to 70: 30 weight ratio, and most preferably 75: 25 weight ratio (citrus peel extract: Guava leaf extract). In the present invention, the weight ratio means a weight ratio of the dried extracts.

The present invention is based on another fact that when the citrus peel extract and Guava leaf extract are incorporated at the above weight mixing ratio, the antiinflammatory effect is surprisingly higher than that of the composition containing other mixing ratios.

More preferably, the present invention provides a composition for anti-inflammatory properties of the present invention containing the citrus peel extract and the guava leaf extract raw dangssuk, aedang mugwort, sacheolssuk (injinho, Artemisia capillaries , warmth ( Artemisia iwayomogi ), white wormwood ( Artemisa sacrorum subsp. Kitamura), hair wormwood ( Artemisa sacrorum subsp. There is manshuria. vestita Kitamura). The present invention provides a pharmaceutical composition for improving or preventing inflammation.

Considering the fact that the anti-inflammatory effect of the composition of the present invention is lowered when the mugwort extract is included in the anti-inflammatory composition containing the citrus peel extract and guava leaf extract according to the present invention, the anti-inflammatory effect of the mugwort extract is high. Lt; / RTI >

In the present invention, "substantially not included" means that the extract contains not more than 5% by weight, preferably not more than 3% by weight, more preferably not more than 1% By weight, preferably not more than 0.5% by weight.

In the present invention, the pharmaceutical composition for treating or preventing inflammation containing the citrus peel extract and Guava leaf extract as an active ingredient comprises (S1) extracting the extract from citrus peel and / or Guava leaf with water or an alcoholic aqueous solution as an extraction solvent step; And (S2) citrus peel extract and guava leaf extract at a weight ratio of 80:20 to 70:30.

Meanwhile, the composition for the treatment or prevention of inflammation prepared by the mixing ratio according to the present invention can be manufactured in the form of pharmaceuticals. Such medicaments may include pharmaceutically acceptable excipients or additives. The extract mixtures of the present invention may be administered alone or in admixture with any convenient vehicle, excipient, etc., and such dosage forms may be single-dose or repeated-dose formulations.

The pharmaceutical product containing the extract mixture of the present invention may be a solid preparation or a liquid preparation. Solid preparations include, but are not limited to, powders, granules, tablets, capsules, suppositories, and the like. Solid form preparations may include, but are not limited to, excipients, flavoring agents, binders, preservatives, disintegrants, lubricants, fillers, and the like. Examples of the liquid preparation include water, a solution such as a solution of propylene glycol, a suspension, an emulsion, and the like, but not limited thereto, and it can be prepared by adding a suitable coloring agent, a flavoring agent, a stabilizer, a tackifying agent and the like.

For example, the acid can be prepared by simply mixing a mixture of citrus peel extract and guava leaf extract prepared at a specific mixing ratio, which is an effective ingredient of the present invention, and pharmaceutically acceptable excipients such as lactose, starch and microcrystalline cellulose. The granule is the above-mentioned extract mixture of the present invention; Suitable excipients which are pharmaceutically acceptable; And a suitable pharmaceutically acceptable binder such as polyvinylpyrrolidone or hydroxypropylcellulose, followed by wet granulation using a solvent such as water, ethanol or isopropanol or dry granulation using a compressive force . Further, the tablet may be prepared by mixing the above granule with a suitable pharmaceutically acceptable salt such as magnesium stearate and then tableting it using a tablet machine.

The extract mixture of the present invention can be administered orally or parenterally depending on the condition to be treated and the condition to be treated, such as an oral preparation, an injection (for example, intramuscular injection, intraperitoneal injection, intravenous injection, infusion, subcutaneous injection, implant), an inhalant, Rectal, rectal, transdermal, topical, and the like, but the present invention is not limited thereto. May be formulated into suitable dosage unit formulations, including those conventionally used and non-toxic, pharmaceutically acceptable carriers, additives, vehicles according to the route of administration.

To achieve the object of improvement of inflammation, the mixture of citrus peel extract and guava leaf extract of the present invention may be administered at a daily dose of about 0.0001 mg / kg to about 10 g / kg, and about 0.001 mg / kg to about 1 g / kg is preferred. However, the dosage may vary depending on the degree of purification of the mixture, the condition of the patient (age, sex, weight, etc.), severity of the condition being treated, and the like. For convenience, the total daily dose may be divided as needed and divided into several doses throughout the day.

The pharmaceutical composition of the present invention is a mixture of active ingredients in citrus peel extract and guava leaf extract to provide an improved anti-inflammatory effect, thus providing therapeutic or preventive use of inflammation. In particular, pharmaceutical compositions composed of optimal mixing ratios are synergistically enhanced to produce an anti-inflammatory effect, which is excellent for the treatment or prevention of inflammation.

Figure 1 is a simplex design configuration for mixture design.
Figure 2 shows the regression and variance analysis for NO production in LPS-induced RAW 264.7 cells.
3 (A) is a triangular-dimensional contour map showing the effect of NO production on the parasitic pea (X1), guava (X2), and phosphorescence (X3) in LPS-induced RAW 264.7 cells. The contour line represents% NO production. Figure 3 (B) is a reaction surface showing the effect of different mixing ratios of parasitic, guava and guinea pig extracts on NO production in LPS-induced RAW 264.7 cells.
Figure 4 shows the regression and variance analysis results for PGE ₂ production in LPS-induced RAW 264.7 cells.
5 (A) is a triangular-dimensional contour plot showing the effect of the parachute rind (X1), guava (X2), and rhinophyse (X3) on PGE ₂ production in LPS-induced RAW 264.7 cells. Contour lines are PGE ₂ Generate%. Figure 5 (B) shows that in LPS-induced RAW 264.7 cells, PGE ₂ It is a reaction surface showing the effect of different mixing ratios of the parchment, guava, and guinea pig extracts on the production.
FIG. 6 is a graph showing the mixing ratios of parasitic pea, guava, and phosphorus, which are the minimum amounts of NO and PGE ₂ production.

Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.

Guava  Manufacture of leaf extract

Guaaba ( Psidium guajava ) was dried and pulverized using a grinder and then passed through a 20 mesh sieve.

The pulverized guava leaves were subjected to reflux extraction for 45 hours at an extraction temperature of 47 캜 using a 20% by weight aqueous solution of 55% ethanol of guava leaves, followed by filtration. The resulting extract was concentrated at 45 캜 or lower on a rotary vacuum evaporator And lyophilized.

Manufacture of Injin-ho extract

Artemisia capillaris Thumb was dried and pulverized using a crusher and then passed through 20 mesh sieve.

The crushed Artemisia leaves were refluxed for 6.5 hours at an extraction temperature of 63 DEG C using a 54% ethanol aqueous solution of 20 times the weight of Artemisia leaves, and then filtered. The extract was concentrated under a rotary vacuum evaporator at 45 DEG C or lower And lyophilized.

Production of Parchmenta pea extract

The supercritical carbon dioxide extraction equipment was used to carry out the extraction of parchmental scales

1) Pearl peel was ground with a household blender and weighed 531.3g. Then, it was placed in a 3L supercritical water extraction tank and the extractor was sealed.

2) Carbon dioxide is injected from the bottom of the extraction tank up to 300 atm using a high pressure gas pump (Haskel Gas Booster, Model No .: AGT-30/75). The internal temperature of the extraction bath is maintained at 50 캜.

3) The extraction valve was opened at the upper part of the extractor, and the high-pressure gas pump was adjusted to maintain the internal pressure and temperature at 300 atm and 50 ° C through the recovery tank.

4) Extraction was carried out for a total of 3 hours, and it was confirmed that the extract was no longer produced. The amount of CO ₂ used was 19 Kg (flow rate: 0.1 Kg / min).

5) 19.77 g of a crude extract containing water was obtained.

Experimental design for optimization of material mixing ratio

Each extract was diluted to a concentration that did not show cytotoxicity through MTT assay and used in the experimental design of the mixture.

The design, data analysis and optimization of the experiment were performed using the statistical software MINITAB ver. 18 and designed according to the mixture design in the experimental design method. As the independent variables, phallic supernatant extract, guava leaf extract, and guinho extract were set as the independent variables. In the mixture experiment, the lower limit was set for the mixing ratio of at least 50% The Simplex Centroid Design was used in the design of the mixture according to the composition ratio of the samples having the constraint, and the setting repetition of the entire design was limited to one. When the total amount of the mixture was set to 1, the limit of each component was restricted as shown in the following equation (1).

Equation (1)

X1 + X2 + X3 = 1

0 = X1 = 0.5

0 = X2 = 1.0

0 = X3 = 1.0

Herein, in the formula (1), X1 represents supercritical extract of parchmental scallion, X2 represents guava leaf extract, and X3 represents aninhinho extract. Ten experimental points were set at the axis and center when each set range was input using the Minitab's mixture design (Fig. 1). Table 1 shows the material mixture ratios of the experiment points. Were performed randomly.

Supercritical extracts of crustacea, crushed guava leaves and crustaceans were mixed at the mixing ratios shown in Table 1 below.

Example Echinacea crassifolia extract Guava leaf extract Injinho extract One 1.0000 0.0000 0.0000 2 0.5000 0.5000 0.0000 3 0.5000 0.0000 0.5000 4 0.7500 0.2500 0.0000 5 0.7500 0.0000 0.2500 6 0.5000 0.2500 0.2500 7 0.6667 0.1667 0.1667 8 0.8333 0.0833 0.0833 9 0.5833 0.3333 0.0833 10 0.5833 0.0833 0.3333

NO production measurement

The NO concentration was determined by measuring the nitrite concentration in the culture medium using the Griess regent. RAW 264.7 cells were plated in 96-well plates at 1 × 10 ⁵ cells / well, cultured for 24 hours, pretreated with each mixture, treated with LPS (lipopolysaccharide) (1 ug / mL) for 2 hours and cultured for 24 hours. The cell culture supernatant (100 uL) was placed in a 96-well plate, and an equal volume of Griess regent was added thereto. After reacting at room temperature for 10 minutes, the absorbance was measured at 540 nm using an ELISA microplate reader. The NO concentration in the culture medium was determined using a standard curve for the concentration of sodium nitrite.

The NO production amount (%) according to the mixing ratio in each of Examples 1 to 10 was measured in the range of 36.00 to 47.58% as shown in Table 2. It was found that the NO extracts of guava leaves and Injinho extracts were lower than those of 45.81 ± 0.62% and 57.79 ± 0.82%, respectively.

As can be seen from the following Table 2, it was found that the mixture of parchment and guava leaves inhibited the NO production more than the mixture of parchment and guinea pigs by comparing Examples 2 and 3 and Examples 4 and 5, respectively. Comparing Examples 2 and 4, it was found that the ratio of 3: 1 to the mixture ratio of parchment and guava leaves was more inhibited than that of 1: 1. Therefore, it is found that the amount of NO produced is decreased as the amount of phosphorus in the mixture is smaller, and the amount of NO production is lower as the ratio of pear pea and guava leaves is closer to 3: 1.

PGE ₂ Production amount measurement

To measure the amount of PGE _2, RAW 264.7 cells were first plated on a 6-well plate at a density of 1 × 10 ⁶ cells / ml, and cultured in a 37, 5% CO ₂ incubator for 24 hours. The cells were pretreated with each mixture, incubated for 2 hours, and cultured for 24 hours with LPS (1 ug / mL). The separated and the cell culture was centrifuged for 3 minutes at 5,000 rpm the supernatant using the PGE ₂ enzyme immunoassay (EIA) kits (R & D Systems, Abington, UK) was measured both PGE _2.

The amount (%) of PGE ₂ produced according to the mixing ratio in each of Examples 1 to 10 was measured in the range of 49.73 to 67.08% as shown in Table 2.

As shown in the following Table 2, when the compositions of Examples 2 and 3 and the compositions of Examples 4 and 5 were compared, it was found that the mixture of Euphrates and Guava leaves inhibited the generation of PGE ₂ more than the mixture of Euphrates and Injinho. Comparing Examples 2 and 4, it was shown that the ratio of 3: 1 to that of 1: 1 mixture of parchment and guava leaves suppressed PGE ₂ production. Therefore, PGE ₂ production will lower the amount of injinho mixture is small, the proportion of pericarp and palsak guava leaves 3: The closer to 1 it can be seen that the PGE ₂ production decreases.

Example In RAW 264.7 cells
LPS-induced NO production In RAW 264.7 cells
The amount of LPS-induced PGE ₂ production One 43.12 63.61 2 40.60 63.98 3 45.35 67.08 4 36.00 49.73 5 46.39 63.98 6 39.78 49.83 7 47.58 52.26 8 38.97 58.16 9 36.29 56.80 10 37.56 57.48

Based on the experimental results shown in Table 2 above, the mixing ratios of the pear pea extract, guava leaf extract and guinea pig extract, which can minimize the NO production amount and the PGE ₂ production amount for the anti-inflammatory activity enhancing effect, were evaluated.

Regression analysis

The analysis of the experimental results was carried out using Minitab 's mixture design analysis. The results of the mixture analysis, the contour diagram and the surface profile of the NO production amount of the parchment, guava and guinea mixture are shown in FIG. 2 and FIG. FIG. 3 (A) is a contour map that visually shows the relationship between the independent variable and the dependent variable calculated from the linear regression equation, and FIG. 3 (B) The position of the optimum condition can be generally known. Fig was a p value of 0.045 in 2 guava palsak * p * value of guava injinho is shown as 0.050 it showed significance at the 5%. In FIG. 3 (A), it was found that as the amount of parchment and guava increased, the amount of NO decreased as the amount of phosphorus decreased. The regression analysis showed that the data fit of the linear model was 90.04% R-squared, and the data were somewhat significant for the linear model. The regression analysis showed a p- value of 0.039 and a linear coefficient of p = 0.078.

PGE ₂ of mixed extract of guinea pea, Guava leaf and Injin-ho The result of the mixture analysis and the surface diagram for the amount of production are shown in FIG. 4 and FIG. 5. In Figure 4, was a p value of 0.011 guava palsak * showed high significance within 5% p value of guava * injinho is shown as 0.006. In Fig. 5 (A), the extract of two of the three components showed a significant effect on the reaction factors. In the case of Paraquat * Guava and Guava * Injin, the reaction factors were different depending on the composition ratio of the two components. Regression analysis showed that the data fit of the linear model showed high reliability with R-squared = 93.07% and regression analysis showed a relatively significant level of p = 0.020.

Optimization of reaction

The Optimization Technique for finding the optimal level of independent factors in the design of a mixture is to find the optimum value of the parameters by finding the response variables for the experimental points at each step and then moving toward the more optimized directions.

In order to predict the optimum mixing ratio of parchment, guava, and phosphorus to minimize the amount of NO and PGE ₂ formation, the mixing ratio was investigated using Minitab Software 's reaction optimization tool. First, in the case of minimizing the NO production amount, the target value of the nitrate characteristic was set to 35.0 based on the results shown in FIG. 3 (A) and Table 2, and the upper limit value of allowable maximum reaction value was set to 48.0. In the case of minimizing the amount of PGE ₂ produced, the target value of the RN characteristic was set at 48.0 and the upper limit of the allowable maximum reaction value was set at 68.0 based on the results of FIG. 5 (A) and Table 2. The weight of NO and PGE ₂ production was set equal to 1, giving equal importance to the target value and the limit value. In the case of importance of NO and PGE ₂ production, the importance of each reaction was set to a default value of 1 because all reactions were equally important.

Result of a reaction Optimizer using the above conditions, the optimal conditions are palsak and blood supercritical extract (X _1), guava leaf extract (X _2), injinho extract (X ₃₎ are respectively 75.9%, 24.1%, 0.0 %. The NO and PGE ₂ production amounts of the mixed extracts were 35.1% and 51.7%, respectively (FIG. 6). The individual satisfaction rate (d) of NO production was 0.99 and the individual satisfaction rate of NO production was 0.81. Overall satisfaction (D) combined with individual satisfaction was 0.90.

Claims (6)

Citrus peel extract and Guava leaf extract as active ingredients,
Wherein the citrus peel extract and guava leaf extract are contained in a weight ratio of 87:13 to 62:38 (citrus peel extract: Guava leaf extract). The pharmaceutical composition for treating or preventing inflammation according to claim 1, wherein the citrus is Citrus hassaku . delete The pharmaceutical composition according to claim 1, wherein the composition comprises citrus peel extract and guava leaf extract in a weight ratio of 80:20 to 70:30 (citrus peel extract: Guava leaf extract). The pharmaceutical composition according to claim 1, wherein the composition contains 5% or less by weight of the mugwort extract. [Claim 5] The pharmaceutical composition according to claim 5, wherein the mugwort extract is an extract of a guinea pig.

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