EXTERNAL COMPOSITION FOR SKIN HAVING ITCHING RELIEVING EFFECT

Abstract

The present invention relates to a skin topical composition having an antipruritic efficacy, comprising a combination of (A) birch sap and (B) betaine, and the skin topical composition does not comprise water added as a separate component. The skin topical composition is a pharmaceutical composition or a cosmetic composition.

Claims

1. Use of a combination of betaine and birch sap in a skin topical composition having an antipruritic efficacy, wherein the skin topical composition does not comprise water added as a separate component.

2. The use according to claim 1, wherein the birch sap is birch sap stock solution.

3. The use according to claim 1, wherein the birch sap is a concentrated birch sap having a concentration degree of 1.05-8 times, preferably 1.1-4 times, more preferably 1.2-2 times.

4. The use according to any one of claims 1-3, wherein the skin topical composition is a pharmaceutical composition or a cosmetic composition.

5. A skin topical composition having an antipruritic efficacy, comprising a combination of (A) birch sap and (B) betaine, wherein the skin topical composition does not comprise water added as a separate component.

6. The composition according to claim 5, wherein the birch sap is birch sap stock solution.

7. The composition according to claim 5, wherein the birch sap is a concentrated birch sap having a concentration degree of 1.05-8 times, preferably 1.1-4 times, more preferably 1.2-2 times.

8. The composition according to any one of claims 5-7, which is a pharmaceutical composition or a cosmetic composition.

9. The composition according to any one of claims 5-8, which comprises 5-99.9% by weight, preferably 20-95% by weight, more preferably 35-90% by weight of birch sap, based on the total weight of the skin topical composition.

10. The composition according to any one of claims 5-9, which comprises 0.01-20% by weight, preferably 0.1-15% by weight, more preferably 1-10% by weight of betaine, based on the total weight of the skin topical composition.

Description

EXAMPLES

[0034] The present invention is further described in detail below with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention in more detail and are not intended to limit the present invention. All similar substitutions and modifications will be apparent to those skilled in the art, and they are all considered to be included in the scope of the present invention.

Example 1: Preparation of Experimental and Control Samples

[0035] In this example, combination samples having the following formulation were prepared, wherein combinations A-1, A-2, and A-3 fall within the present invention, and combinations B-E are controls.

TABLE-US-00001 Comb*. A-1 Comb. A-2 Comb. A-3 Comb. B Comb. C Comb. D Comb. E (wt %) (wt %) (wt %) (wt %) (wt %) (wt %) (wt %) Water 0 0 0 0 50 90 50 Betaine 10 10 10 0 10 10 0 Birch sap 0 0 90 100 40 0 50 stock solution 1.3 times 90 0 0 0 0 0 0 concentrated birch sap 3.5 times 0 90 0 0 0 0 0 concentrated birch sap *comb. = combination

[0036] The preparation procedure of the above formulations was as follows: birch sap stock solution, concentrated birch sap or water were heated to 80° C., added with betaine, and stirred uniformly for later use. The resulting samples were all colorless and transparent solutions.

Example 2: Effects of Experimental and Control Samples on Histamine Release from RBL-2H3 Cells

[0037] The effects of samples A-E as prepared above on the release of histamine from RBL-2H3 cells were tested according to the following test methods.

[0038] Experimental principle: Compound 48/80 (a polymer produced by condensation of N-methyl-p-methoxyphenylethylamine and formaldehyde) is a tool medicine to induce mast cell degranulation, the mechanism is that it acts on mast cell membrane and induces an increase of intracellular calcium ions to change the amount of second messengers cAMP and cGMP, resulting in mast cell degranulation and histamine release.

[0039] Experimental materials: RBL-2H3 cell line was provided by the Animal Center of Zhejiang Academy of Medical Sciences; the test samples included above seven samples combinations; the reagents included fetal bovine serum, Tyrode's salt, Compound 48/80, and histamine Elisa kit.

[0040] Test Steps:

TABLE-US-00002 Preparation 1.0 mg of 48/80 powder was weighed and diluted with 1 of Com- mL of Tyrode's solution, and 50 uL of the dilution pound was diluted to 1000 uL, i.e. Compound 48/80 solution 48/80 at a concentration of 50 ug/mL, and use solution solution right after it was ready. Preparation One part of Tyrode's salt was diluted with 80 mL of of sterilized deionized water and volumed to 100 mL, Tyrode's thereby preparing Tyrode's solution of 10 times solution concentration, and autoclave sterilized. Cell RBL-2H3 cells in logarithmic growth phase were treatment spread to the bottom of a culture flask and digested with a digestive solution containing 0.25% trypsin and 0.03% EDTA, centrifuged at 1200 rpm for 5 mins; resuspended with 1 times Tyrode's solution and counted, and diluted to 1 × 10.sup.4/mL; centrifuged again at 1200 rpm for 5 mins to remove the supernatant. Sample Model group: 700 uL of cell suspension was centrifuged grouping and and resuspended with Tyrode's solution, held at experiments 37° C. for 30 mins; sample group: 700 uL of cell suspension was centrifuged and resuspended with sample solution, held at 37° C. for 30 mins; the samples were mixed well and dispensed into 3 tubes, 200 uL per tube; the model group and the 7 sample groups: 50 uL of Compound 48/80 solution was added to the cell suspension to reach a final concentration of 10 ug/mL, and held at 37° C. for 20 mins. 3 replicates in each group. Histamine The above groups of samples were cooled in an ice box for determi- 10 mins to stop the reaction. The samples after the reaction nation were centrifuged (4° C., 1500 rpm, 30 mins) to precipitate the cells at the bottom of EP tube, and the supernatants were transferred to new centrifuge tubes. The precipitated cells were resuspended with 250 μL of 1 times Tyrode's solution, heated in a 90° C. water bath, taken out after 5 mins and immediately placed in an ice box, after the samples were completely cooled, they were taken out and heated again in a 90° C. water bath, and frozen-thawed repeatedly several times to break cells. The supernatants and the cell solutions were diluted 10 times, and the stock solutions were stored in a refrigerator at −20° C. The diluted samples were determined for histamine using an imported histamine kit. The histamine release was calculated using the following formula: histamine release (%) = extracellular histamine release amount/(extracellular histamine release amount + intracellular histamine amount) × 100%

[0041] Experimental results: The experimental results are summarized in Table 1 below.

TABLE-US-00003 TABLE 1 Test results of histamine release in cell model P value P value (vs. Release % (vs. model) Combination A-1) Model group 72.7 ± 3.2 — 0.000 Combination A-1 30.9 ± 1.1 0.000 — Combination A-2 36.3 ± 1.6 0.000 0.003 Combination A-3 40.4 ± 1.0 0.000 0.009 Combination B 45.1 ± 2.9 0.000 0.004 Combination C 51.8 ± 3.9 0.001 0.001 Combination D 57.5 ± 4.7 0.004 0.001 Combination E 59.9 ± 2.8 0.003 0.000

[0042] The above results of the cell experiments showed that combinations A-1, A-2 and A-3 within the scope of the present invention significantly reduced histamine release amount as compared to control combinations B-E. This showed that the combination of birch sap and betaine or the combination of concentrated birch sap and betaine had synergistic effects on inhibition of histamine release, and the combination of 1.3 times concentrated birch sap and betaine showed further better effects.

Example 3: Effect of Experimental and Control Samples on Release of Interleukin-4 (IL-4) from Mast Cells

[0043] According to the following test method, the effects of the seven samples A-E as prepared above on the release of IL-4 from mast cells were tested.

[0044] Experimental principle: The specific antigen is crosslinked with the IgE antibody bound to the cell membrane, resulting in cell activation and rapid secretion of inflammatory mediators related to granulation (histamine, IL-4, IL-13, etc.). In addition to immune triggering, the degranulation reaction of mast cells can be caused by non-specific triggering agents such as anti-IgE, Compound 48/80, etc., these agents exhibit characteristics similar to those that rely on IgE release mediators. In this study, Compound 48/80 was used to stimulate mast cells to degranulate and release the inflammatory mediator IL-4 to investigate the protective efficacy of the test samples on the allergic reaction of mast cells.

[0045] Experimental materials: Mast cells were provided by the Animal Center of Zhejiang Academy of Medical Sciences. Reagents included fetal bovine serum, RPM1640 medium, Tyrode's salt, Compound 48/80, sodium cromoglycate, and Elisa (interleukin-4) kit. The test samples included control group, model group, positive drug (sodium cromoglycate) group, and above seven samples A-E.

[0046] Experimental method: Each of samples was prepared using RPMI1640 medium, and each of samples was provided with 8 duplicate wells. Mast cells were inoculated into a 96-well culture plate at a density of 5×10.sup.6/mL, 90 μL per well, and added with 10 μL/well of a corresponding sample; cultured in a 5% CO.sub.2 incubator at 37° C., and taken out after 4 h; in addition to the control group, the other groups were added with a stimulus 48/80 (final concentration 20 mg/L), and continued to incubate in an incubator at 37° C. for 2 h, and then centrifuged at 1500 rpm for 5 mins to get the supernatant, and the content of IL-4 in the supernatant was detected by Elisa (interleukin-4) kit.

[0047] Experimental results: The experimental results are summarized in Table 2 below.

TABLE-US-00004 TABLE 2 Test results of IL-4 release from cell models Release % P value(vs model) Control group 34.4 ± 7.9  0.000 Model group 67.6 ± 9.9  — Positive drug group 44.2 ± 12.3 0.003 Combination A-1 50.8 ± 14.7 0.014 Combination A-2 52.9 ± 15.7 0.035 Combination A-3 53.2 ± 11.3 0.031 Combination B 55.7 ± 12.5 0.043 Combination C 57.6 ± 15.0 0.083 Combination D 61.4 ± 12.1 0.109 Combination E 61.6 ± 9.1  0.132

[0048] The results showed that the combinations A-1, A-2 and A-3 within the scope of the present invention all significantly inhibited (p<0.05) IL-4 release caused by Compound 48/80 stimulating mast cells. This showed that the combination of birch sap and betaine or the combination of concentrated birch sap and betaine has a synergistic effect on the inhibition of IL-4 release, and the combination of 1.3 times concentrated birch sap and betaine showed further better effects.

Example 4: Preparation of Spray Composition Having Antipruritic Efficacy

[0049] This example provided a spray composition having an antipruritic efficacy, and its formulation was as follows:

TABLE-US-00005 Amount Ingredients (wt %) Birch sap 77.75 Betaine 5 Glycerin 5 Glycerin polyether-26 0.5 Phytosteryl/octyldodecyl lauroyl glutamate 1 Butanediol 5 Pentanediol 2 Glycerol triethylhexanoate 2 PEG-60 hydrogenated castor oil 1 Citric acid 0.1 Arginine 0.1 Methyl hydroxybenzoate 0.15 Phenoxyethanol 0.4

[0050] The above spray composition was prepared as follows:

1. Birch sap was heated to 80° C., added with betaine, citric acid, glycerin, methyl hydroxybenzoate, butanedio and pentanediol in turn and mixed, stirred to dissolve uniformly;
2. Phytosteryl/octyldodecyl lauroyl glutamate, glycerol trimethylhexanoate, glycerin polyether-26, and PEG-60 hydrogenated castor oil were heated to 80° C., stirred uniformly, and then added with the mixture obtained in 1;
3. The resultant was cooled to 40° C., added with phenoxyethanol, and then the pH was adjusted with arginine, and discharged.

[0051] Efficacy test: 30 patients over 40 years old with pruritus senilis, male or female, only itch and no skin lesions were selected. The above spray composition was applied to the itch site after cleaning every morning and evening. Before and 4 weeks after the use of the composition, self-evaluation was carried out for itch and was ranked into significant improvement, obvious improvement, certain improvement, no significant improvement, and no improvement depending on the degree of improvement. The results showed that 27 patients (90%) had a significant improvement in itch after using the product, while the other 3 patients also had certain improvement. This showed that the spray composition had an excellent antipruritic efficacy.

Example 5: Preparation of Lotion Composition Having Antipruritic Efficacy

[0052] This example provided a lotion composition having an antipruritic efficacy, and its formulation was as follows:

TABLE-US-00006 Amount Ingredients (wt %) Birch sap 76.7 Betaine 6 Allantoin 0.4 Panthenol 0.6 Glycerin 5 Hydrolyzed sodium hyaluronate 0.5 Pentanediol 3 Caprylic/capric triglycerides 3 Butyrospermum parkii (shea butter) 1 Carbomer 0.1 Xanthan gum 0.1 Cetyl alcohol 1 Glycerol stearate/PEG-100 stearate 1 Glycerol triethylhexanoate 1 Phenoxyethanol 0.4 Methyl hydroxybenzoate 0.1 Arginine 0.1

[0053] The above lotion composition was prepared as follows:

1. Aqueous phase: birch sap, glycerin, betaine, panthenol, allantoin, hydrolyzed sodium hyaluronate, pentanediol, xanthan gum, carbomer and methyl hydroxybenzoate were mixed and heated to 80° C., stirred to dissolve uniformly;
2. Oil phase: cetyl alcohol, glycerol stearate/PEG-100 stearate, caprylic/capric triglyceride, glycerol methylhexanoate, butyrospermum parkii (shea butter). The above raw materials were heated to 80° C., and stirred to dissolve uniformly;
3. The aqueous phase and the oil phase were mixed, homogenized and emulsified for 5 mins. Upon the completion of the emulsification, arginine was added, stirred for 15 mins, cooled to 40° C., and phenoxyethanol was added.

[0054] Efficacy test: 30 patients over 40 years old with pruritus, male or female, only itch and no skin lesions were selected. The above lotion composition was applied to the itch site after cleaning every morning and evening. Before and 4 weeks after the use of the composition, self-evaluation was carried out for itch and was ranked into significant improvement, obvious improvement, certain improvement, no significant improvement, and no improvement depending on the degree of improvement. The results showed that 28 patients (93.3%) had a significant improvement in itch after using the product, while the other 2 patients also had certain improvement. This showed that the lotion composition had an excellent antipruritic efficacy.

Example 6: Preparation of Body Milk Composition Having Antipruritic Efficacy

[0055] This example provided a body milk composition having an antipruritic efficacy, and its formulation was as follows:

TABLE-US-00007 Lotion A Ingredients (wt %) Birch sap 70.03 Betaine 3 Allantoin 0.1 Panthenol 0.6 Glycerin 8 Hydrolyzed sodium hyaluronate 0.1 Trehalose 2 Polymethylsilsesquioxane 1 Myristyl alcohol 1 Beeswax 0.2 Stearyl alcohol 0.3 Stearic acid 0.5 Polydimethylsiloxane 1 Nicotinamide 0.3 Triethanolamine 0.2 Acrylates/C10-30 alkyl acrylate crosspolymer 0.15 Dipropylene glycol 3 Caprylic/capric triglycerides 3 Butyrospermum parkii (shea butter) 2 Carbomer 0.1 Xanthan gum 0.1 Hydrogenated lecithin 0.3 Glycerol stearate/PEG-100 stearate 1 Caprylyl glycol 0.02 Phenoxyethanol 0.4 Methyl hydroxybenzoate 0.1 Avena Sativa extract 1.5

[0056] The above body milk composition was prepared as follows:

1. Aqueous phase: birch sap, glycerin, betaine, panthenol, allantoin, hydrolyzed sodium hyaluronate, dipropylene glycol, trehalose, hydrogenated lecithin, xanthan gum, carbomer, acrylates/C10-30 alkyl acrylate crosspolymer and methyl hydroxybenzoate were mixed and heated to 80° C., stirred to dissolve uniformly;
2. Oil phase: glycerol stearate/PEG-100 stearate, caprylic/capric triglyceride, polymethylsilsesquioxane, myristyl alcohol, beeswax, stearyl alcohol, stearic acid, butyrospermum parkii (shea butter), polydimethylsiloxane. The above raw materials were heated to 80° C., stirred to dissolve uniformly;
3. The aqueous phase and the oil phase were mixed, homogenized and emulsified for 5 mins. Upon the completion of the emulsification, triethanolamine was added, stirred for 15 mins, cooled to 40° C., and phenoxyethanol, Avena Sativa extract, caprylyl glycol and nicotinamide were added.

[0057] Efficacy test: 30 patients over 40 years old with pruritus, male or female, only itch and no skin lesions were selected. The above lotion composition was applied to the itch site after cleaning every morning and evening. Before and 4 weeks after the use of the composition, self-evaluation was carried out for itch and was ranked into significant improvement, obvious improvement, certain improvement, no significant improvement, and no improvement depending on the degree of improvement. The results showed that 29 patients (96.7%) had a significant improvement in itch after using the product, while the other 1 patient also had an obvious improvement. This showed that the body milk composition had an excellent antipruritic efficacy.

Example 7: Preparation of Cream Composition Having Antipruritic Efficacy

[0058] This example provided a cream composition having an antipruritic efficacy, and its formulation was as follows:

TABLE-US-00008 Amount Ingredients (wt %) Birch sap 76.95 Betaine 3 Allantoin 0.1 Panthenol 0.5 Glycerin 5 Hydrolyzed sodium hyaluronate 0.5 Glycerin polyether-26 2 Xanthan gum 0.1 Glyceryl caprylate/caprate 2 Butyrospermum parkii (shea butter) 1 Cetyl alcohol 1 Glycerol stearate/PEG-100 stearate 1.5 Pentaerythrityl tetraethylhexanoate 3 Polydimethylsiloxane 2 Methyl hydroxybenzoate 0.2 Propyl hydroxybenzoate 0.1 Carbomer 0.3 Phenoxyethanol 0.4 Caprylyl glycol 0.05 Arginine 0.3

[0059] The above cream composition was prepared as follows:

1. Aqueous phase: birch sap, betaine, glycerin, panthenol, allantoin, hydrolyzed sodium hyaluronate, glycerin polyether-26, xanthan gum, carbomer, methyl hydroxybenzoate;
2. Oil phase: cetyl alcohol, glyceryl caprylate/caprate, pentaerythrityl tetraethylhexanoate, glycerol stearate/PEG-100 stearate, butyrospermum parkii (shea butter, polydimethylsiloxane, propyl hydroxybenzoate. The above raw materials were heated to 80° C., stirred to dissolve uniformly;
3. The aqueous phase and the oil phase were mixed, homogenized and emulsified for 5 mins. Upon the completion of the emulsification, Arginine was added, stirred for 15 mins, cooled to 40° C., and phenoxyethanol and caprylyl glycol were added in turn.

[0060] Efficacy test: 30 patients over 40 years old with pruritus, male or female, only itch and no skin lesions were selected. The above lotion composition was applied to the itch site after cleaning every morning and evening. Before and 4 weeks after the use of the composition, self-evaluation was carried out for itch and was ranked into significant improvement, obvious improvement, certain improvement, no significant improvement, and no improvement depending on the degree of improvement. The results showed that 28 patients (93.3%) had a significant improvement in itch after using the product, while the other 2 patient also had an obvious improvement. This showed that the cream lotion composition had an excellent antipruritic efficacy.

[0061] The technical solutions of the above-mentioned examples are preferred embodiments of the present invention, and do not constitute a limitation to the scope of the appended claims. Various modifications and variations can be made without departing from the spirit and principle of the present invention, and these modifications and variations should also be considered within the scope of the present invention.