KR20170000740A

KR20170000740A - Soluble microneedle patch for lignin peroxidase delivery

Info

Publication number: KR20170000740A
Application number: KR1020150162716A
Authority: KR
Inventors: 김정아; 심우선; 강내규
Original assignee: 주식회사 엘지생활건강
Priority date: 2015-06-24
Filing date: 2015-11-19
Publication date: 2017-01-03

Abstract

The present invention relates to a skin administration system of lignin peroxidase which can exert an excellent effect by stably impregnating and effectively delivering lignin peroxidase into skin, and is a microneedle including lignin peroxidase. The present invention also provides a skin administration method of lignin peroxidase using the micro-needle.

Description

Soluble microneedle patch for lignin peroxidase delivery for lignin peroxidase delivery

The present invention relates to a microneedle and a method for producing the microneedle.

In general, microneedle is used for delivery of active substances such as drugs and vaccines in vivo, detection of biological analyte, and biopsy. The delivery of the pharmacologically or cosmetically active ingredient using microneedles is intended for the delivery of the active substance through the skin, not the vascular system such as blood vessels or lymphatic vessels.

Examples of the method using the microneedle include a method of forming a predetermined number of holes or more on the skin using a microneedle device such as a roller with a microneedle attached thereto, (Active ingredient) is coated on the skin to allow the active ingredient to be administered simultaneously with the perforation of the skin.

Unlike color cosmetics, which generally act on the surface of skin in cosmetic preparations and exhibit their effects, the effects of wool, hair growth, hair loss prevention, wrinkle improvement, whitening, skin moisturizing, hair removal, skin nutrition, etc., It works inside the skin such as transdermal and dermis and shows its effect.

On the other hand, lignin peroxidase is an enzyme obtained from a fermentation microorganism called phanerochaete chrysosporium and has been known to exhibit excellent whitening effects such as skin tone, skin texture improvement and gloss enhancement (Korean Patent Laid- -2005-0088381).

However, lignin peroxidase has problems such as formulation stability and possibility of inducing skin irritation.

Korean Patent Publication No. 10-2005-0088381

SUMMARY OF THE INVENTION The object of the present invention is to provide a system for effectively delivering lignin peroxidase into the skin by stably impregnating a lignin peroxidase in a soluble micro needle, a method for producing such a system, and a system for producing lignin peroxidase Or a pharmaceutically acceptable salt thereof.

Accordingly, the present invention provides a soluble microneedle comprising lignin peroxidase.

The present invention also provides a method for producing a microneedle comprising lignin peroxidase.

The present invention also provides a microneedle patch comprising the microneedles.

In addition, the present invention provides a cosmetic skin administration method of lignin peroxidase which effectively transfers lignin peroxidase into skin, wherein the micro needle is applied.

In order to solve the above-mentioned problems, the present invention provides a dissolvable microneedle comprising lignin peroxidase, more preferably, the material forming the microneedle is dissolved in the skin, The fine needle dissolves or collapses to release lignin peroxidase into the skin.

The inventors of the present invention have confirmed that the lignin peroxidase can be effectively delivered into the skin by impregnating the lignin peroxidase in the soluble micro-needle after a long period of research, and the instability of the formulation including lignin peroxidase can be solved Thereby completing the present invention. More specifically, impregnating lignin peroxidase in a soluble micro-needle to overcome the problem of discoloration and tearing caused by having a stabilized formulation at a low pH, directly acting inside the skin to alleviate skin irritation, Was more effective than the others.

As used herein, the term "skin irritation mitigation" refers to prevention, mitigation, and treatment of inflammatory irritation such as itching, tingling, burning, and erythema caused by skin irritants, .

The present inventors have studied various administration systems and none of the systems has been able to simultaneously solve various problems of the lignin peroxidase. The inventors of the present invention have surprisingly succeeded in solving the problem of discoloration and deterioration due to unstable formulation of lignin peroxidase by effectively impregnating micro needle with lignin peroxidase and effectively delivering it to the skin.

As used herein, the term "impregnation" may refer to a form in which the lignin peroxidase can be contained in the microneedles, and preferably includes i) a lignin peroxidase in combination with a solubilizing material that forms microneedles, or ii) A lignin peroxidase may be contained in the hole by puncturing the fine needle. When microneedles are prepared in the form of i) and ii), lignin peroxidase can penetrate effectively into the skin, and particularly when preparing microneedles in the form of i), the problem of instability of the lignin peroxidase .

Therefore, it is most preferable that the material forming the microneedles is dissolved in the living body so that the lignin peroxidase contained in the microneedles is effectively released into the skin.

In a preferred embodiment of the present invention, the material forming the microneedle is dissolved in the skin, and when the microneedles are applied to the skin, the microneedles are dissolved or collapsed, whereby the lignin peroxidase contained in the microneedles effectively penetrates into the skin can do.

In the present invention, the microneedles are preferably soluble in the skin. For example, hyaluronic acid, sodium carboxymethyl cellulose (Na-CMC), or the like may be used to form a soluble microneedle. , Water-soluble polymers such as vinylpyrrolidone-vinyl acetate copolymer, polyvinyl alcohol, and polyvinyl pyrrolidone; Sugars such as Xylose, Sucrose, Maltose, Lactose and Trehalose; Or a mixture thereof may be used. In consideration of the stability of the lignin peroxidase and the compatibility with the lignin peroxidase, taking into consideration the skin permeation intensity of the micro-needle and the dissolution rate in the skin comprehensively, A mixture of hyaluronic acid, sodium carboxymethyl cellulose (Na-CMC), and saccharide (preferably trehalose) is preferable as a substance, and a mixture of glycerin Is more preferable.

Preferably, the microneedles of the present invention may further include plasticizers, surfactants, preservatives, anti-inflammatory agents, etc. in addition to the above-mentioned components for forming microneedles.

Examples of the plasticizer include polyols such as ethylene glycol, propylene glycol, dipropylene glycols, butylene glycol, and glycerin. Or may be used in combination.

The length of the microneedles according to the present invention is not limited to a specific size.

In the present invention, the lignin peroxidase may be contained in an amount of 0.001 to 10% by weight, preferably 0.01 to 5% by weight, and more preferably 0.1 to 3% by weight based on the total weight of the microneedles. If the content is less than 0.001% by weight, the effective effect may not be exhibited. If the content is more than 10% by weight, the possibility of skin irritation may be high rather than the excellent effect.

In the present invention, the lignin peroxidase may be stably impregnated with the lignin peroxidase so as not to discolor or remove during the production of the microneedles. Conventionally, in the cream formulations to be applied to the skin, it has been necessary to stabilize the lipid peroxidase to include a specific component in order to contain the sensitive lignin peroxidase. However, in the present invention, impregnation into the microneedles has solved such a problem.

When the microneedles according to the present invention are applied to the skin, the lignin peroxidase can be significantly improved in skin permeation compared to other skin administration systems.

In addition, the microneedles according to the present invention can be used for skin whitening purposes.

The present invention also provides a microneedle patch comprising the microneedles, and more particularly, a microneedle patch for administering (or delivering) the microneedle attached lignin peroxidase.

In the present invention, the patch may be a sheet prepared by attaching one or more micro-needles impregnated with the lignin peroxidase of the present invention and having the micro-needle attached thereto. The size of the sheet is not limited to a specific size, but can be appropriately adjusted depending on the amount of the lignin peroxidase to be absorbed on the skin or the attachment site. In addition, one or more, preferably a plurality of micro-needles may be attached to the surface of the sheet which can be attached to the skin.

In addition, lignin peroxidase may be added to the surface of the patch which can be adhered to the skin, so that the lignin peroxidase can penetrate into the hole formed by the microneedles. In order to solve the problem of discoloration and detachment of lignin peroxidase, Preferably, the peroxidase is impregnated with a microneedle.

The present invention also provides a method of preparing a microcapsule comprising: S1) filling a mold with a material forming a microneedle including a lignin peroxidase and an intracutaneous solubility material; And S2) heating, drying, and then separating the mold, and then lignin peroxidase.

The method may be prepared by impregnating a microneedle with lignin peroxidase, and the impregnation may include, but is not limited to, i) lignin peroxidase in combination with i) an intracutaminous solubilizing agent forming microneedles, ii) ) Containing a lignin peroxidase in the hole by puncturing the fine needle, and the like.

In order to impregnate the lignin peroxidase into the microneedles, the microneedle forming material and the lignin peroxidase are mixed together in a mold for microneedle formation, and then the mold is heated, dried and separated So that a fine needle can be produced.

In the present invention, the in-skin solubility material may include, for example, hyaluronic acid, sodium carboxymethyl cellulose (Na-CMC), vinyl pyrrolidone-vinyl acetate copolymer, polyvinyl alcohol vinyl alcohol, and polyvinyl pyrrolidone; Sugars such as Xylose, Sucrose, Maltose, Lactose and Trehalose; Or a mixture thereof may be used.

In addition, the present invention provides a cosmetic skin administration method of lignin peroxidase, which effectively transfers lignin peroxidase into the skin, characterized by applying the microneedles according to the present invention.

The term "application" is used herein to include both attaching a microneedle according to the present invention to the skin or attaching a microneedle patch according to the present invention to the skin.

The present invention provides a method for preparing a system for administration of lignin peroxidase which can exhibit an excellent effect through effective skin transfer of lignin peroxidase. The present invention also provides a skin administration method of lignin peroxidase, characterized by applying such microneedles.

BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments of the invention and, together with the description of the invention, Should not be construed as limited.
FIG. 1 is a view showing one example of various methods for manufacturing a microneedle according to the present invention.
FIG. 2 shows a Franz diffusion cell for evaluating the drug release behavior of the microneedles according to the present invention.
3 is a graph showing the skin permeation rate of lignin peroxidase of Example 1 and Comparative Example 1. Fig. (Red: Example 1, blue: Comparative Example 1)
4 is a graph showing the effect of improving the skin color of the microneedles according to the present invention.

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.

&Lt; Preparation of soluble fine needle >

Soluble microneedles are prepared by solution casting method, and they are prepared by casting solution into mold, filling solution into micro mold by vacuum or centrifugation and drying.

Conventional synthetic and natural water-soluble polymers were used for the materials forming the microneedle structure.

&Lt; Preparation of soluble micro-needle containing lignin peroxidase (Example 1) >

ingredient Example 1
(Unit: wt%) oligo-HA 6 Na-CMC 6 Trehalose 10 Glycerin 5 Lignin peroxidase 2 Citric acid Suitable amount water To 100

After dissolving oligo-HA (Hyaluronic acid), Na-CMC (Sodium carboxymethyl cellulose), Trehalose, Glycerine citric acid and lignin peroxidase in purified water, 4.5. The prepared solution was cast in a silicon microneedle mold and then centrifuged at 3000 rm for 10 minutes to fill the micro mold with the solution. After the solution was filled, it was dried in a drying oven (70 ° C) for 3 hours, and the fine needle was separated from the silicone mold using an adhesive film (Example 1).

&Lt; Preparation of lignin peroxidase-containing cream (Comparative Example 1) >

Raw material name Comparative Example 1
(Unit: wt%) One Cetearyl alcohol 2.5 2 Polyglyceryl-3 methyl glucoside distearate 2 3 Glyceryl stearate 2 4 Hydrogenated polydecene 5 5 Cyclohexasiloxane 3 6 Purified water To 100 7 glycerin 5 8 Dipropylene glycol 5 9 1,2-hexanediol 2 10 Carbomer Suitable amount 11 Polyacrylate Crosspolymer-6 0.5 12 Xanthan gum 0.3 13 Lignin peroxidase 2 14 Citric acid Suitable amount

To compare the lignin peroxidase impregnated with the microneedles and the lignin peroxidase applied to the oil-in-water cream, a cream having a pH of 4 to 4.5 and containing lignin peroxidase was prepared.

The raw materials 1 to 5 and the raw materials 6 to 14 were heated and melted at 75 ° C to 80 ° C as shown in Table 2, and the mixture was stirred at 5500 rpm for 2 minutes and then cooled to produce a homogeneous mixture (Comparative Example 1).

< Experimental Example 1>

Confirmed over time stability

Example 1 and Comparative Example 1 were stored at a constant temperature of 0 ° C, room temperature, 40 ° C, and 50 ° C for 4 weeks to confirm the stability of sample precipitation and discoloration.

division 30 days Room temperature 40 ℃ 50 ℃ 0 ℃ Example 1 stability stability Non-discolored stability Comparative Example 1 discoloration Discoloration Discoloration stability

As can be seen from the results of Table 3, the results of Example 1 were good at both the low temperature and the high temperature as compared with Comparative Example 1.

< Experimental Example 2>

Skin irritation improvement effect

The present inventors observed the degree of skin irritation (N = 20) by attaching the microneedles for 1 hour to the inside of the arm and the microneedles for 2 hours on the inside of the arms, respectively.

The reason why the skin irritation of Example 1 is smaller than that of Comparative Example 1 is that it is because the cream is applied for a long time in actual use, but the contact time of the skin is short because the microneedles are removed after a short time (1 hour) .

division After 1 day Two days later After 4 days After 6 days After 8 days After 10 days After 12 days After 14 days Example 1 0.05 0.05 0.08 0.08 0.08 0.05 0.10 0.08 Comparative Example 1 0.05 0.10 0.15 0.30 0.28 0.20 0.25 0.26

* Skin irritation is judged to be irritation when the skin irritation index is 0.2 or more, and skin irritation is usually 0.05 to 0.1 in the case of functional cosmetic.

< Experimental Example 3>

Drug release behavior

The present inventors measured the lignin peroxidase content of porcine skin tissue and the acceptor solution with time using Liquid Chromatography using a Franz diffusion cell. Example 1 was applied to pig skin and Comparative Example 1 was applied (time: 2 hours, temperature: 32 ° C) to penetrate the pig skin and dissolve, and then the microneedles and cream were removed.

Pig skin absorbed with lignin peroxidase by Example 1 and Comparative Example 1 was placed in a franz diffusion cell (Fig. 2), and skin permeation amount of lignin peroxidase was compared with time.

As a result, as shown in FIG. 3, the amount of the lignin peroxidase impregnated in the fine needle was directly transmitted to the skin by the needle, and the permeation amount thereof was about 25 μg or more, about 25 times And the skin permeability was higher than that of the cream.

< Experimental Example 4>

Color difference system Melanin index measurement

The present inventors evaluated the degree of skin color improvement by measuring the melanin index using a colorimeter (MPA-580), treating the melanin index at each time point The results are shown in Table 5 and FIG. (N = 20)

As a result, an improvement of about 2 times as much as that of Example 1 was obtained as compared with Comparative Example 1, and it was confirmed that lignin peroxidase effectively penetrated into skin by microneedles and skin whitening effect was high, As a result of the melanin index analysis, it was confirmed that the test group using Example 1 had a lower melanin index than the test group using Comparative Example 1.

　
　 The melanin index (N = 20) 0 weeks 1 week 2 weeks 4 weeks 6 weeks Comparative Example 1 215.63 203.98 198.99 194.57 185.53 Example 1 211.04 196.21 191.96 182.22 170.45

Claims

Soluble microneedles containing lignin peroxidase.

The method according to claim 1,
Wherein the material forming the microneedle is dissolved in the skin.

3. The method of claim 2,
The microneedle forming material may be selected from the group consisting of hyaluronic acid, sodium-carboxymethylcellulose, sodium carboxymethyl cellulose (Na-CMC), vinylpyrrolidone-vinyl acetate copolymer, polyvinyl alcohol Poly vinyl alcohol, polyvinyl pyrrolidone, saccharides, or a mixture of two or more thereof.

The method of claim 3,
Wherein the microneedles further comprise a plasticizer in addition to the material forming the microneedles.

The method according to claim 1,
Wherein the lignin peroxidase is contained in an amount of 0.001 to 10% by weight based on the total weight of the microneedles.

The method according to claim 1,
Wherein the microneedles improve the skin permeability of the lignin peroxidase.

The method according to claim 1,
Wherein the microneedles are for skin whitening.

A microneedle patch comprising a microneedle according to any one of claims 1 to 7.

S1) filling a mold with a substance which forms a microneedle comprising a lignin peroxidase and an intracutaneously soluble substance, and mixing the same; And
S2) heating the mold, drying and then separating the lignin peroxidase.

A method for the cosmetic skin administration of a lignin peroxidase which transfers a lignin peroxidase into the skin, characterized by applying a microneedle according to any one of claims 1 to 7.