A TOPICAL COMPOSITION COMPRISING AN EXTRACT OF COMBINED HERBS COMPRISING LONGANAE ARILLUS FOR TLSP INHIBITION AND THE TREATMENT OR ALLEVIATION OF SKIN INFLAMMATORY DISEASE AND THE USE THEREOF

Abstract

The present invention is related to a topical pharmaceutical composition and cosmetic composition comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix as an active ingredient to inhibit TLSP (thymic stromal lymphopoietin) cytokine expression or to treat and alleviate skin inflammatory diseases.

Claims

1. A topical pharmaceutical composition comprising a combined herb extract of Longanae Arillus Ligustici Tenuissimi Rhizoma and Polygalae radix as an active ingredient to inhibit TLSP (thymic stromal lymphopoietin) cytokine expression or to treat and alleviate skin inflammatory diseases.

2. The topical pharmaceutical composition according to claim 1, wherein said combined herb extract is (a) the combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix with the mixed ratio based on the dried weight of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix (w/w) ranging from 0.01-100:0.01-100:0.01-100 weight part (w/w), or (b) the combination of each extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix with the mixed ratio based on the dried weight of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix (w/w) ranging from 0.01-100:0.01-100:0.01-100 weight part (w/w).

3. The topical pharmaceutical composition according to claim 1, wherein said extract is extracted with at least one solvent selected from water, C 1-C4 lower alkyl alcohol such as methanol, ethanol, propanol, butanol, etc, acetone, ethyl acetate, chloroform, hexane, butyleneglycol, propyleneglycol or glycerin.

4. The topical pharmaceutical composition according to claim 1, wherein said skin inflammatory diseases is the disease selected from group of pruritus caused by aging or atopy, chronic relapsing dermatitis such as atopic dermatitis, psoriasis, and the like; contact dermatitis, seborrheic dermatitis, neurodermatitis, xeroderma, erythema, inflammatory dermatitis, psoriasis, or atopic disease.

5. A method of inhibiting TLSP (thymic stromal lymphopoietin) cytokine, treating or alleviating skin inflammatory diseases in a mammal comprising topically administering to said mammal an effective amount of the combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix and pharmaceutically acceptable carrier thereof.

6. (canceled)

7. (canceled)

8. A cosmetic composition comprising the combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix as an active ingredient in an amount effective to inhibit TLSP (thymic stromal lymphopoietin) cytokine or to treat and alleviate skin inflammatory diseases.

9. The cosmetic composition according to claim 8, wherein said extract is extracted with at least one solvent selected from water, C1-C4 lower alkyl alcohol such as methanol, ethanol, propanol, butanol, etc, acetone, ethyl acetate, chloroform, hexane, butyleneglycol, propyleneglycol or glycerin.

10. The cosmetic composition according to claim 8, wherein said skin inflammatory diseases is the disease selected from group of pruritus caused by aging or atopy, chronic relapsing dermatitis such as atopic dermatitis, psoriasis, and the like; contact dermatitis, seborrheic dermatitis, neurodermatitis, xeroderma, erythema, inflammatory dermatitis, psoriasis, or atopic disease.

11. The cosmetic composition according to claim 8, wherein said composition is a form selected from skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, moisture cream, hand cream, foundation, essence, nutrient essence, pack, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, treatment, beauty solution and the like.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0089] FIG. 1 shows the dermatitis induced dorsal skin lesions treated with test sample (WIN), dexamethasone (DEX) and distilled water (DIW);

[0090] FIG. 2 shows the stained test results with H&E and toluidine blue (TB) on the dermatitis induced dorsal skin lesions treated with test sample (WIN), dexamethasone (DEX) and distilled water (DIW)

BEST MODE FOR CARRYING OUT THE INVENTION

[0091] It will be apparent to those skilled in the art that various modifications and variations can be made in the compositions, use and preparations of the present invention without departing from the spirit or scope of the invention.

[0092] The present invention is more specifically explained by the following examples.

[0093] However, it should be understood that the present invention is not limited to these examples in any manner.

Examples

[0094] The following Examples and Experimental Examples are intended to further illustrate the present invention without limiting its scope.

Example 1. The Preparation of Inventive Combined Extract (1)

[0095] 20 g of dried Longanae Arillus (Buyoung Yakup Co. Ltd.), 20 g of dried Ligustici Tenuissimi Rhizoma (Buyoung Yakup Co. Ltd.) and 20 g of dried Polygalae radix(Buyoung Yakup Co. Ltd.) were cut into small pieces, mixed with 6 fold volume (v/w) of 20% ethanol in water and the mixture was subjected to reflux extraction at 90±5° C. for 3 days. After filtration of the extract through filter paper (pore size, less than 10 μm) to remove the debris, the remaining debris was further extracted two times with 4 fold volume (v/w) of 20% ethanol in water and the extract was filtered with filter paper (pore size, less than 10 μm).

[0096] The collected extract was mixed with together and concentrated under vaccuo (16 21 brix) to afford concentrated extract. The concentrated extract was dried with freeze drying process and pulverized (less than 50 mesh) to obtain 20.5 g (powder as dried basis, yield 33.4%) of inventive combined extract (1) (designated as “WIN-1001X” hereinafter)

Example 2-6. The Preparation of Inventive Combined Extract (2)-(6)

[0097] Excepting adopting different combined ratio as well as different solvents disclosed in Example 1, all the procedure was identical with those in Example 1 to obtain various inventive combined extract of Longanae Arillus (LA), Ligustici Tenuissimi Rhizoma (LT) and Polygalae radix (PR) i.e., inventive combined extract (2) to inventive combined extract (6) of the present invention, which are used as a test samples in following experiment.

TABLE-US-00001 TABLE 1 various kinds of combined extract Sample weight (g) Extract Final Example LA* PR* LT* solvent* name weight yield Example 2 10 5 50 10% EtOH WIN-1002X 16.6 g 25.6% Example 3 20 50 5 Water WIN-1003X 24.7 g 32.9% Example 4 10 80 20 70% BuOH WIN-1004X 32.3 g 29.4% Example 5 5 50 20 50% EtOH WIN-1005X 21.5 g 28.7% Example 6 30 10 2 hexane WIN-1006X 12.6 g 30.1% *Longanae Arillus (LA), Ligustici Tenuissimi Rhizoma (LT), Polygalae radix (PR)

Experimental Example 1. Inhibitory Effect on Cytokine Expression (In Vitro)

[0098] In order to determine the anti-inflammatory activity of inventive extract, following inhibition test of cytokine expression using HaCaT cell was performed according to the procedure disclosed in the literature (Jeong et al., 2019, J. Invest. Dermatol., May; 139 (5): pp 1098-1109).

[0099] HaCaT cell (human epithelial keratinocyte cell, 300493, CLS) was inoculated into DMEM medium containing 10% Fetal bovine serum, 100 units/m1 of penicillin, 100n/m1 of streptomycin (D6429, Sigma-Aldrich Co. Ltd) and was incubated in the incubator (HERA cell 150i, Thermo Fisher Scientific Co. Ltd.) maintaining optimum humidity (85-95%) and 5% CO.sub.2 atmosphere.

[0100] For performing gene expression test, the incubated cells were transferred to 12 wells and 50 ng/ml of TNF alpha (RC214-12, Biobasic Co. Ltd) was treated therewith for 1 hour to induce inflammatory response. Dexamethasone (200 nM, positive control, “DEX”, D4902, Sigma-Aldrich Co. Ltd.) and distilled water (negative control, “DIW”) were used as comparative controls.

[0101] 1 hour after inducing the inflammation, 1 μg/ml of inventive extract prepared in Examples was treated with identical medium and subjected to incubation for 1 hour. After the incubation, RNA (FATRR-001, Favorgen) was extracted from the cell and cDNA was synthesized from the RNA using by cDNA synthesis kit (RRO36A, TAKARA). The polymerization reaction was performed using by the synthesized cDNA and Sybrgreen kit (RT500M, Enzynomics) and then Real-time-PCR was performed using by primers for various cytokines involved in skin inflammation (RPLPO, TSLP, GM-CSF and IL-1beta) as disclosed in Table 2.

TABLE-US-00002 TABLE 2 The used primers in RT-PCR method Sequence human* direction sequence I. D RPLP0 forward 5′- AGC CCA GAA CAC TGG TCT C-3′ 1 reverse 5′- ACT CAG GAT TTC AAT GGT GCC-3′ 2 TSLP forward 5′-TAT GAG TGG GAC CAA AAG TAC CG-3′ 3 reverse 5′-GGG ATT GAA GGT TAG GCT CTG G-3′ 4 GM-CSF forward 5′-TCC TGA ACC TGA GTA GAG ACA C-3′ 5 reverse 5′-TGC TGC TTG TAG TGG CTG G-3′ 6 IL-1β forward 5′-CTC CAG GGA CAG GAT ATG GA-3′ 7 reverse 5′-TCT TTC AAC ACG CAG GAC AG-3′ 8 *abbreviation- RPLP0 (Ribosomal Protein Lateral Stalk Subunit P0); TSLP (thymic stromal lymphopoietin); GM(Granulocyte-macrophage)-CSF (colony stimulating factor); IL (interleukin)

[0102] As can be seen in Table 3 showing quantitative result of the RT-PCR, the test sample group treated with the inventive extract, sharply inhibited the expressed level of various cytokine involved in skin inflammation comparing with negative control group treated with distilled water (DIW) and it has been confirmed that the inhibitory activity of the test sample on the expression of various cytokine involved in skin inflammation is equivalent to that of positive control group treated with dexamethasone (DEX).

[0103] Accordingly, it has been confirmed that the various kind of inventive combined extract prepared in Examples 1-6 have potent inhibitory effect on skin inflammation.

TABLE-US-00003 TABLE 3 Inhibition effect on cytokine expression TSLP — TNFα TNFα TNFα TNFα TNFα TNFα — DIW WIN-1001X WIN-1002X WIN-1003X WIN-1005X Dex 1 132.4692 47.43735 60.85783 48.9323 55.34286 52.49334 0.462769 26.91228 9.645089 24.95619 19.85678 26.59252 11.2336 GM-CSF — TNFα TNFα TNFα TNFα TNFα TNFα — DIW WIN-1001X WIN-1002X WIN-1003X WIN-1005X Dex 1 4.473627 1.982161 2.069408 2.384771 1.917569 1.935997 0.111 0.817826 0.889233 0.326074 0.871501 0.599711 0.581338 IL-1β — TNFα TNFα TNFα TNFα TNFα TNFα — DIW WIN-1001X WIN-1002X WIN-1003X WIN-1005X Dex 1 4.152715 1.407169 1.437399 2.064964 1.662578 1.080503 0.483565 1.087056 0.394622 0.1926 0.620225 0.193175 0.413136

Experimental Example 2. Inhibitory Effect on Atopic Dermatitis (In Vivo)

[0104] To confirm the inhibitory effect of inventive extract on the atopic dermatitis, the animal model test using by mice, was performed according to the method disclosed in the reference (Li et al., Drug. Des. Devel. Ther., 2016, Feb. 19; 10:781-191).

[0105] 100 μL of 0.15% (w/v) DNFB (2,4-dinitrofluorobenzene, D1529, Sigma-Aldrich Co. Ltd.) was spread on abdominal cavity of 6 weeks-old BALB/C female mouse (DBL Co., Ltd. Incheon, Korea) and after removing the central dorsal hair, 100 μL of 0.15% (w/v) DNFB (2,4-dinitrofluorobenzene, D1529, Sigma-Aldrich Co. Ltd.) was spread at every third day starting from at 7th day to 16th day to induce skin inflammation. The test sample (10 mg/ml of inventive extract WIN-1001X, prepared in Examples) as well as Dexamethasone (200 μM, positive control, “DEX”, D4902, Sigma-Aldrich Co. Ltd.) and distilled water (negative control, “DIW”) used as comparative controls were spread every starting from at 7th day to 16.sup.th day of initial treatment day of DNFB.

[0106] For the purpose of comparing with test group, the identical test excepting using acetone instead of DNFB and DIW instead of test sample with the above procedure was performed.

[0107] As can be seen in FIG. 1, it can be seen that skin dermatitis occurred on the dorsal skin of mice at 16th day after the sample treatment and the severity of skin dermatitis has been classified into four scores, i.e., score 0 (no syndrome), 1 (mere dermatitis), 2 (mean dermatitis) and 4 (severe dermatitis) with respect to four criteria on the syndrome, i.e., (i) erythema/bleeding, (ii) edema, (iii) abrasion/maceration, and (iv) scar/dryness to be total score 12.

[0108] As can be seen in FIG. 1 and Table 4, it has been confirmed that the inventive combined extract showed potent improving effect on skin dermatitis comparing with negative control group, of which effect is similar to that of positive control group treated with dexamethasone (DEX).

TABLE-US-00004 TABLE 4 Improving effect on skin dermatitis Acetone DNFB DNFB DNFB DIW DIW WIN-1001X DEX 0 8.8 5 2.2 0 1.30384 0.707107 0.83666

[0109] Additionally, the histological analysis on skin tissue of test animal has been performed.

[0110] Specifically, the dorsal skin tissue occurring dermatitis was fixed on 4% (w/v) paraformaldehyde solution (P6148, Sigma-Aldrich) in shaker maintaining 4° C. (CR300, FINEPCR) at 16th day after the sample treatment and 12 hours after the fixation, the dehydrated and transparent tissue was embedded in paraffin, sliced (5 μm of width) to make tissue slices.

[0111] The tissue slices were stained with H&E staining agent for staining skin epidermal layer {hematoxylin (S3309<DAKO) & Eosin (109844, Millipore)} or toluidine blue for staining mast cells infiltrated into inflamed lesion (TB, 185426, Sigma-Aldrich) and the result was observed using by microscope (EVOS XL., Life Technologies).

[0112] As can be seen FIG. 2 and Tables 5-6, it has been confirmed that the test group treated with inventive combined extract showed significantly reduced width of skin epidermal layer as well as reduced number of mast cell infiltrated into inflamed lesion comparing with negative control group treated with DIW, of which effect is similar to that of positive control group treated with dexamethasone (DEX).

TABLE-US-00005 TABLE 5 effect on the relative area of epidermis (fold) Acetone DNFB DNFB DNFB DIW DIW WIN-1001X DEX 1 7.06432606 3.55861484 1.638117 0.173221 2.07300369 0.42330352 0.875806

TABLE-US-00006 TABLE 6 effect on the relative number of mast cells (fold) Acetone DNFB DNFB DNFB DIW DIW WIN-1001X DEX 1 3.182796 1.72043 0.854839 0.312754 1.003359 0.263386 0.270532

Experimental Example 3. Inhibitory Effect on Cytokine Expression (In Vivo)

[0113] In order to determine the anti-inflammatory activity of inventive extract, following inhibition test of cytokine expression using test animal was performed according to the procedure disclosed in the literature (Li et al., Drug. Des. Devel. Ther., 2016, Feb. 19; 10:781-191).

[0114] At 16th day after the treatment of DNFB, RNA (FATRR-001, Favorgan) was extracted from the dorsal skin of mice prepared in Experimental Example 2 and cDNA was synthesized from the RNA using by cDNA synthesis kit (RRO36A, TAKARA). The polymerization reaction was performed using by the synthesized cDNA and Sybrgreen kit (RT500M, Enzynomics) and then Real-time-PCR was performed using by primers for various cytokines involved in skin inflammation (GADPH, TSLP, GM-CSF, IL-4, IL-10, IL-13, IL-31 and IL-33) as disclosed in Table 7.

TABLE-US-00007 TABLE 7 The used primers in RT-PCR method Sequence mouse* direction sequence I. D GAPDH forward 5′- AGG TCG GTG TGA ACG GAT TTG-3′ 9 reverse 5′-TGT AGA CCA TGT AGT TGA GGT CA-3′ 10 TSLP forward 5′-AGC TTG TCT CCT GAA AAT CGA G-3′ 11 reverse 5′-AGG TTT GAT TCA GGC AGA TG TT-3′ 12 CSF2 forward 5′-AGG GTC TAC GGG GCA ATT TC-3′ 13 reverse 5′-TCA CAG TCC GTT TCC GGA GTT-3′ 14 IL-4 forward 5′-GGT CTC AAC CCC CAG CTA GT-3′ 15 reverse 5′-GCC GAT GAT CTC TCT CAA GTG AT-3′ 16 IL-10 forward 5′-GCT CTT ACT GAC TGG CAT GAG-3′ 17 reverse 5′-CGC AGC TCT AGG AGC ATG TG-3′ 18 IL-13 forward 5′-CCT GGC TCT TGC TTG CCT T-3′ 19 reverse 5′-GGT CTT GTG TGA TGT TGC TCA-3′ 20 IL-22 forward 5′-ATG AGT TTT TCC CTT ATG GGG AC-3′ 21 reverse 5′-GCT GGA AGT TGG ACA CCT CAA-3′ 22 IL-31 forward 5′-TCA GCA GAC GAA TCA ATA CAG C-3′ 23 reverse 5′-TCG CTC AAC ACT TTG ACT TTC T-3′ 24 IL-33 forward 5′-GCT GCA GAA GGG AGA AAT CAC G-3′ 25 reverse 5′-GGA GTT GGA ATA CTT CAT TCT AGG 26 TCT CAT-3′ *abbreviation- GAPDH (Glyceraldehyde-3-phosphate dehydrogenase); TSLP (thymic stromal lymphopoietin); GM(Granulocyte-macrophage)-CSF (colony stimulating factor); IL (interleukin)

[0115] As can be seen in Table 8 showing quantitative result of the RT-PCR, the test sample group treated with the inventive extract, sharply inhibited the expressed level of various cytokine involved in skin inflammation comparing with negative control group treated with distilled water (DIW) and it has been confirmed that the inhibitory activity of the test sample on the expression of various cytokine involved in skin inflammation is equivalent to that of positive control group treated with dexamethasone (DEX).

[0116] Accordingly, it has been confirmed that the inventive combined extract prepared in Example has potent inhibitory effect on skin inflammation.

TABLE-US-00008 TABLE 8 Inhibition effect on cytokine expression CSF2 Acetone DNFB* DNFB DNFB DIW DIW** WIN-1001X DEX*** 1 72.14648 4.676082 7.817781 0.210396 12.96696 1.723294 0.98556 IL-4 Acetone DNFB DNFB DNFB DIW DIW WIN-1001X DEX 1 11.36284 1.06316 4.798023 0.543902 3.453585 0.037212 2.312695 IL-10 Acetone DNFB DNFB DNFB DIW DIW WIN-1001X DEX 1 19.29307 8.810186 20.44472 0.354329 1.973736 0.411741 4.157348 IL-13 Acetone DNFB DNFB DNFB DIW DIW WIN-1001X DEX 1 48.62991 6.488924 12.56024 0.465986 10.37666 3.599525 8.866739 IL-22 Acetone DNFB DNFB DNFB DIW DIW WIN-1001X DEX 1 29.18521 1.303359 13.08191 0.577444 2.801416 0.465745 1.945029 IL-31 Acetone DNFB DNFB DNFB DIW DIW WIN-1001X DEX 1 3.933601 1.217765 1.000609 0.473134 0.44744 0.304217 0.13203 IL-33 Acetone DNFB DNFB DNFB DIW DIW WIN-1001X DEX 1 9.471517 0.541047 0.736505 0.487178 2.797898 0.460806 0.486004 *DNFB: 2,4-dinitrofluorobenzene; **DIW: distilled water; ***DEX: Dexamethasone

Experimental Example 4. Inhibitory Effect on TSLP Cytokine Expression (In Vivo)

[0117] In order to determine the anti-inflammatory activity of inventive extract, following inhibition test of TSLP cytokine expression using test animal was performed according to the procedure disclosed in the literature (Li et al., Drug. Des. Devel. Ther., 2016, Feb. 19; 10:781-191).

[0118] At 16th day after the treatment of DNFB, RNA (FATRR-001, Favorgan) was extracted from the dorsal skin of mice prepared in Experimental Example 2 and cDNA was synthesized from the RNA using by cDNA synthesis kit (RRO36A, TAKARA). The polymerization reaction was performed using by the synthesized cDNA and Sybrgreen kit (RT500M, Enzynomics) and then Real-time-PCR was performed using by primers for TSLP cytokines involved in skin inflammation as disclosed in Table 7.

[0119] As can be seen in Table 9 showing quantitative result of the RT-PCR, the test sample group treated with the inventive extract, sharply inhibited the expressed level of TSLP cytokine involved in skin inflammation comparing with negative control group treated with distilled water (DIW) and it has been confirmed that the inhibitory activity of the test sample on the expression of TSLP cytokine involved in skin inflammation is equivalent to that of positive control group treated with dexamethasone (DEX).

[0120] Accordingly, it has been confirmed that the inventive combined extract prepared in Example has potent inhibitory effect on the expression of TSLP cytokine.

TABLE-US-00009 TABLE 9 Inhibition effect on TSLP cytokine expression TSLP Acetone DNFB DNFB DNFB DIW DIW WIN-1001X DEX 1 7.514841 1.525431 1.310709 0.45284 1.954331 0.784039 0.32691

[0121] Statistics Analysis

[0122] The average and standard error were calculated from the test results obtained from the experiment. The significance difference test was analyzed using t-test, and the significance level (P-value) was expressed as P≤0.05=*, P≤0.01=**, and P≤0.001=***.

MODE FOR THE INVENTION

[0123] Hereinafter, the formulating methods and kinds of excipients will be described, but the present invention is not limited to them. The representative preparation examples were described as follows.

[0124] Preparation of Skin Lotion

[0125] Extract of Example (WIN-1001X) 1.00%

[0126] Glycerol 3.00%

[0127] Ethanol 1.00%

[0128] Propylene glycol 0.10%

[0129] Flavour trace amount

[0130] Distilled water up to 100%

[0131] Skin preparation was prepared by dissolving the active components according to conventional lotion preparation method.

[0132] Preparation of Lotion

[0133] Extract of Example (WIN-1002X) 3.00%

[0134] L-ascorbic acid-2-magnesium phosphate 1.00%

[0135] Soluble collagen (1% solution) 1.00%

[0136] Sodium citric acid 0.10%

[0137] 1,3-butylene glycol 3.00%

[0138] Distilled water up to 100%

[0139] Lotion preparation was prepared by dissolving the active components according to conventional lotion preparation method.

[0140] Preparation of Cream

[0141] Extract of Example (WIN-1003X) 3.00%

[0142] Polyethyleneglycomonosterate 2.00%

[0143] Monostearate glycerin 1.00%

[0144] Cetyl alcohol 4.00%

[0145] Squalene 6.00%

[0146] Tri 2-glycerly ethylhexanoate 6.00%

[0147] Sphingo-glycolipid 1.00%

[0148] 1,3-butylene glycol 3.00%

[0149] Distilled water up to 100%

[0150] Cream preparation was prepared by dissolving the active components according to conventional cream preparation method.

[0151] Preparation of Pack

[0152] Extract of Example (WIN-1004X) 5.00%

[0153] Polyvinyl alcohol 13.00%

[0154] L-ascorbic acid-2-magnesium phosphate 1.00%

[0155] Lauroylhydroxyproline 1.00%

[0156] Soluble collagen (1% solution) 2.00%

[0157] 1,3-butylene glycol 3.00%

[0158] Ethanol 5.00%

[0159] Distilled water up to 100%

[0160] Sugar 20 g

[0161] Fructose 20 g

[0162] Lemon flavor optimum amount

[0163] Distilled water 100 ml

[0164] Pack preparation was prepared by dissolving the active components according to conventional pack preparation method.

[0165] Preparation of Beauty Solution

[0166] Extract of Example (WIN-1005X) 2.00%

[0167] Hydroxyethylene cellulose (2% solution) 12.00%

[0168] Xanthin gum (2% solution) 2.00%

[0169] 1,3-butylene glycol 3.00%

[0170] Glycerin concentration 4.00%

[0171] Sodium hyaluronte 5.00%

[0172] Distilled water 100 ml

[0173] Beauty solution preparation was prepared by dissolving the active components according to conventional beauty solution preparation method

[0174] The invention being thus described as will be obvious that it may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the present invention, and all such modifications as would be obvious to those skilled in art are intended to be included within the scope of the following claims.

[0175] The invention being thus described, it will be obvious that the same may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the present invention, and all such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.

INDUSTRIAL APPLICABILITY

[0176] As described in the present invention, the present invention provides a topical composition and cosmetic composition comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix, the present inventors demonstrated that the anti-inflammatory effects of inventive combined composition is potent by accomplishing in vitro experiments such as the inhibitory test on the expression of cytokines involved in skin inflammation (RPLPO, TSLP, GM-CSF and IL-1beta) (Experimental Example 1); as well as in vivo experiments such as the inhibitory effect on the atopic dermatitis using by BALB/C mice (Experimental Example 2); the inhibition test of expression of various cytokines involved in skin inflammation (GADPH, TSLP, GM-CSF, IL-4, IL-10, IL-13, IL-31 and IL-33) using test animal (Experimental Example 3); the inhibition test of TSLP cytokine expression using test animal (Experimental Example 4), therefore, it is confirmed that inventive combined extract is very useful in the alleviation or treatment of skin inflammation as a form of topical medicament or cosmetic composition.