KR102099326B1 - Soluble Microneedle for polyphenol delivery - Google Patents

Abstract

The present invention relates to a microneedle containing polyphenols, and to a skin administration system of polyphenols having a wrinkle improvement effect. The present invention can increase the skin delivery rate of polyphenols. The present invention provides a microneedle for skin administration of polyphenol having an effect of improving skin wrinkles.

Description

Soluble microneedle for polyphenol delivery {Soluble Microneedle for polyphenol delivery}

The present invention relates to a soluble microneedle, and to a soluble microneedle containing a polyphenol excellent in wrinkle improvement effect. More specifically, the present invention provides a skin penetration system for polyphenols using the soluble microneedle.

Drug delivery through the skin has been used in various fields and forms due to its ease of use. Drugs that pass through these skins are mainly intended to be delivered to the systemic circulation through the skin, but in addition, drugs such as atopy treatments, whitening or wrinkle improvement cosmetics are also used for the purpose of delivering to the organs of the skin itself. . Despite this convenience and functionality, there are many difficulties in delivering the drug through the skin due to the structure of the skin, so it is not easy to develop a drug that passes through the skin. The stratum corneum of the skin is a brick structure composed of keratin-rich keratinocytes and a mortar filled with lipids such as ceramide, fatty acid, or wax between the keratinocytes. Structure. This structure acts as a barrier and has very low material permeability. Only low molecular structure components of 500 Da or less can be delivered into the skin by a diffusion method, and only substances with excellent lipid affinity can pass through the skin.

Meanwhile, the release of the neurotransmitter is caused by the formation of a passage between the two boundaries after the synaptic vesicle and the presynaptic membrane located at the nerve terminal containing the neurotransmitter are fused. The three protein complex SNARE protein, consisting of the vesicle protein VAMP (synaptobrevin) and the plasma membrane binding protein, Syntaxin 1a, SNAP-25, provides a fundamental force for the fusion of synaptic vesicles and synaptic membranes. Here, the neurotransmitter release channel is opened by the membrane fusion between the synaptic vesicle and the synaptic membrane, t-SNARE and vesicle (a complex of two kinds of syntacsin 1a protein and SNAP-25 protein attached to the target membrane) vesicle) is the result of the operation of v-SNARE attached to. When the membrane is fused, rearrangement of the lipid bilayer occurs. Since the biofilms strongly push each other, these membranes do not spontaneously fuse, and a strong force from outside must overcome the repulsive force between the membranes, and it is known that the SNARE protein provides this force. Thus, the formation of the SNARE complex is a key phenomenon of exocytosis, including the release of neurotransmitters.

Recently, there is an effort to apply the cosmetic type of the vegetable or the actual botox by using the fact that the plant polyphenols exert similar effects to botox by controlling the release of neurotransmitters, but most polyphenols Since the solubility is not good, the amount that can be contained in the cosmetic formulation is limited, and accordingly, the skin permeation amount is very low, and thus there is a problem that it is difficult to exhibit efficacy.

Therefore, the problem to be solved by the present invention is to provide a soluble microneedle impregnated with a polyphenol component having excellent skin permeability.

By blocking nerve transmission, impregnating the soluble microneedle with polyphenol, which has excellent wrinkle improvement effect, is applied to the skin and penetrated into the skin without pain by the microneedle, and the microneedle is dissolved by moisture in the skin and delivers the drug into the skin To provide a soluble microneedle.

The present invention is to solve the problem of limited solubility of the polyphenols due to poor solubility in the general cosmetic composition, and to provide a skin delivery system that can be applied to the skin by improving the solubility of the polyphenols.

In order to solve the above problems, the present invention provides a microneedle comprising a polyphenol or a derivative thereof. Preferably, the present invention provides a microneedle containing polyphenols and having a wrinkle improving effect. The material forming the microneedle may be dissolved or biodegradable in the skin, and when the skin is applied, the microneedle is dissolved or disintegrated to release polyphenol, and the polyphenol can be stably delivered to the skin.

The present inventors have studied various administration systems, and as for any system, as mentioned above, polyphenols have low solubility, which limits their application by applying only to general cosmetics and applying to the skin. The present inventors made a surprising invention that after several efforts, by containing polyphenols in a soluble microneedle in the skin, it can effectively reduce the transmission of neurotransmitters and show wrinkle improvement effects.

To achieve the above task, the microneedle must be soluble in the skin, and to form a soluble microneedle hyaluronic acid, sodium-carboxymethyl cellulose (Na-CMC, sodium carboxymethyl cellulose), vinyl skin Water-soluble polymers such as lollydon-vinyl acetate copolymer, poly vinyl alcohol, and poly vinyl pyrrolidone; Sugars such as xylose, sucrose, maltose, lactose, and trehalose; Or mixtures of these can be used. In particular, considering the skin penetration strength of the microneedle and the dissolution rate in the skin comprehensively, hyaluronic acid, sodium-carboxymethyl cellulose (Na-CMC, sodium carboxymethyl cellulose), and saccharide (saccharide) ) (More preferably, a mixture of Trehalose) is more preferable, and even more preferably mixed with Glycerine (described below). Preferably, the microneedle according to the present invention may additionally include a plasticizer, a surfactant, a preservative, an anti-inflammatory agent, etc. in addition to the above-mentioned components forming the microneedle.

As the plasticizer, for example, polyols such as ethylene glycol, propylene glycol, dipropylene glycol, butylene glycol, and glycerine are solely used. It can be used as or mixed.

Preferably, the microneedle of the present invention regulates neurotransmitter discharge, and contains polyphenols having a wrinkle improvement effect in an amount of 0.01 to 10% by weight based on the total weight of the microneedle, more preferably 0.1 to 5% by weight have.

The microneedle may contain a high content of a polyphenol component, and may provide a polyphenol skin delivery system in which the amount delivered to the skin is significantly increased compared to a formulation of a general cosmetic composition.

 The polyphenol component has poor solubility, and thus the amount contained in the cosmetic composition is extremely small, but the microneedle of the present invention has a very low water content and can be delivered to the skin by applying a high content without precipitation of the polyphenol component.

The polyphenols are camperol, quercetin, myricetin, luteolin, delchinidine, cyanidin, butane, ellagic acid, ampelopsin, hespyridine, aurantinidin, europinidin, pella It may be any one or more selected from the group consisting of pelagindin, malvidin, peonidin, petunidin, and rosinidin and derivatives thereof. May be any one or more selected from the group consisting of ampelopsin, hesperidin or derivatives thereof.

In the present specification, the derivatives of the polyphenols are free form (free acid or base form) and all prodrugs, polymorphs, hydrates, solvates, tautomers, and sterics Unless otherwise specified, such as isomers (stereoisomer), includes all pharmaceutically acceptable forms as described above, and was used to include all active forms of the compound. According to an embodiment of the present invention, a microneedle containing polyphenols inhibits SNARE complex formation, thereby inhibiting membrane fusion and, as a result, inhibiting the release of neurotransmitters, for example, acetylcholine, muscle cells. Can suppress the contraction.

In addition, the present invention provides a microneedle patch system for administration (for delivery) of a polyphenol or a derivative thereof to which the microneedle is attached.

The microneedle of the present invention can be manufactured by a method for manufacturing a conventional soluble microneedle, and the manufacturing method is not particularly limited.

The present invention also provides a method for skin administration of polyphenol with improved solubility and improved skin delivery rate, characterized by using a microneedle according to the present invention.

In addition, the present invention provides a use for improving wrinkles of a microneedle containing a polyphenol or a derivative thereof having the effect of improving wrinkles and controlling wrinkles.

According to an embodiment of the present invention, the present invention provides a microneedle containing polyphenols or derivatives thereof, and inhibiting SNARE complex formation.

The present invention provides a microneedle having an effect of improving skin wrinkles.

The present invention provides a microneedle for skin administration, which can improve the skin penetration efficiency of polyphenols or derivatives thereof having a wrinkle improving effect. The present invention also provides a method for skin administration of a polyphenol or a derivative thereof, characterized by using such a microneedle.

The following drawings attached to the present specification are intended to illustrate preferred embodiments of the present invention, and serve to further understand the technical idea of the present invention together with the contents of the above-described invention, so the present invention is limited to those described in those drawings. It should not be interpreted as limited.
1 is a view showing an example of a number of methods for manufacturing a microneedle according to the present invention.
Figure 2 shows a Franz diffusion cell for evaluating the drug release behavior of the microneedle according to the present invention.
3 exemplarily shows a polyphenol having a botox-like effect of the present invention. Myricetin, delchinidine, cyanidine, camperol, quercetin, picetin, butane, luteolin, ellagic acid, epigallocatechin gallate (EGCG), ampelopsin, hesperidine and derivatives thereof can be used.
Figure 4 shows the results of the SNARE formation inhibition of camphorol, myricetin, and ampelopsin. (Cell line: C2C12 (muscle cell) + NG108-15 (nerve cell) condition, concentration: ppm)
5 is a graph showing the results of comparing the content of ampelopsin in pig skin and acceptor solution.
6 is a graph showing the results of confirming the wrinkle improvement effect of the ampelopsin cream and ampelopsin microneedle on the human body.
Figure 7 shows the results confirming the effect of inhibiting the muscle contraction of camphorol, myricetin, and ampelopsin.

Hereinafter, examples and the like will be described in detail to help understanding of the present invention. However, the embodiments according to the present invention can be modified in many different forms, and the scope of the present invention should not be construed as being limited to the following examples. The embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art. The contents described in this specification are expressed as weight% unless otherwise specified.

<Production of Soluble Micro Needle>

Soluble Micro Needle is manufactured by solution casting method, and the solution is cast into a mold, filled with a liquid in a vacuum or centrifuge, and then dried. At this time, the general synthetic and natural water-soluble polymers are used to form the micro needle structure. .

<Manufacture of SNARE-forming polyphenol microneedle>

A microneedle having the composition of Table 1 below was prepared. After dissolving Hyaluronic acid (Oligo-HA ™), Sodium carboxymethyl cellulose (Na-CMC) and Trehalose in purified water, Glycerin, PEG-40 hydrogenated castor oil (HCO-40 ™) and ampelopsin solution (Ampelopropsin 10%, DPG 90 %) Was added to prepare an ampelopsin solution (DPG: Dipropylene glycol). After casting the prepared polyphenol dispersion solution into a Silicone Micro Needle mold, centrifugation was performed at 3,000 rpm for 10 minutes to fill the micro mold with the liquid.

After filling the solution, dried in a drying oven (70 ° C.) for 3 hours, the micro needle was separated from the silicone mold using an adhesive film. The contents of Table 1 below are expressed in weight percent.

The manufacturing process is shown in FIG. 1.

Polyphenol MN oligo-HA 6 Na-CMC 6 Trehalose 10 Glycerin 5 HCO-40 0.2 Ampropoxin-DPG solution (10%) 1.5 water To 100

<Preparation of SNARE-forming polyphenol oil-in-water cream>

In order to compare the skin permeation amount of the polyphenols impregnated in the microneedle, as a comparative example, a general water-in-oil emulsifier type cream was impregnated and compared.

ingredient Formulation example C14-22 alcohol, C12-20 alkyl glucoside
(Mixture C14-22 alcohol: C12-20 alkyl glucoside = 80: 20 weight ratio)
Glyceryl stearate, PEG-100 stearate
(50:50 weight ratio of the mixture)
Glyceryl stearate
Cetearyl alcohol
Polyglyceryl-3 methyl glucose distearate
Hydrogenated polydecene
Cyclohexasiloxane
Carbomer
Tromethamine
glycerin
DPG
1,2-hexanediol
Ampelopsin
Purified water 1.5

1.2

0.9
1.5
1.5
4.5
3.5
0.2
0.2
3
5
2
0.5
Residual amount (To 100)

<Drug release behavior>

Ampelopin release from the microneedle prepared above was evaluated using a Franz diffusion cell equipped with pig skin (see FIG. 2). A PBS solution containing 30% by weight of DPG was used as an aqueous solution.

That is, with the Franz diffusion cell, the ampeloxin content of pig skin tissue and acceptor solution over time was measured using Liquid Chromatography.

Although the amount of the cream containing ampelopsin penetrated through the skin was minimal, the ampelopsin impregnated in the micro needle penetrated directly into the skin by the needle, and the amount of penetration was higher than that of the cream.

The amount of cream containing ampoplopin is about 1 ug that penetrates through the skin, but the amount is negligible, but the polyphenol impregnated in the microneedle penetrates directly into the skin by the needle, and its penetration is 13 ug or more, which is about 13 times higher than the cream The amount of skin permeation was shown. The results are shown in FIG. 5.

<Wrinkle improvement effect>

Ampheplopsine cream and Ampelopsin-impregnated Microneedle were treated daily for 12 weeks on the wrinkles around the eyes, and the degree of wrinkle improvement was confirmed by a silicone replica and an image analysis method of wrinkles. (N = 20)

Compared to ampoplopsin cream, it showed more than 5 times better improvement effect in the microneeedle impregnated with ampelopsin, which was confirmed that the effect of wrinkle improvement was high because the ampelopsin effectively penetrated into the skin by the microneedle. The results are shown in FIG. 6.

<Inhibition experiment of SNARE complex formation>

In order to confirm the inhibition of SNARE complex formation of polyphenols, such as ampelopsin, camperol, and myricetin, SDS-PAGE (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) analysis was performed. It was confirmed that the complex formed when the SNARE proteins SNAP25, SynH3, and Vps proteins were mixed at a molar concentration of 1: 1: 1, was inhibited by adding a polyphenol compound at a concentration of 1-50 ppm. After dropping each protein into a 1 ml tube, a polyphenol compound was added to the mixture, followed by reaction at room temperature for 30 minutes. Thereafter, electrophoresis was performed on 12% SDS-PAGE to confirm whether SNARE complex was formed.

It was confirmed that camphorol, myricetin, and ampelopsin all effectively inhibit the SNARE complex in a concentration-dependent manner. As a result of the experiment, it was confirmed that the ability to inhibit SNARE complex formation was active at lower concentrations of camphorol and ampelopsin than myricetin. The effect of inhibiting SNARE formation of myricetin, ampelopcin (dihydroxymycetin), camperol, and A440 was confirmed and shown in FIG. 4. (Cell line: C2C12 (muscle cell) + NG108-15 (nerve cell) condition, concentration: ppm)

<Experiment of muscle contraction>

C2C12 cells were cultured in DMEM medium containing 10% fetal calf serum and 1% antibiotic on a plate. Thereafter, neuroblasts were additionally co-cultured on the same plate. Thereafter, when the C2C12 cell contraction starts, the number of C2C12 cell contractions is measured for 30 seconds, and after removing all the medium, washed three times with PBS, and the medium containing no calf serum and 50 ppm of polyphenol compound are reacted for 2 hours. Ordered. Afterwards, the number of contractions of C2C12 cells was measured for another 30 seconds to confirm the degree of muscle contraction inhibition.

It was confirmed that the results of reducing the number of contractions of C2C12 cells by inhibiting the release of neurotransmitters from camphorol, myricetin, and ampelopsin from neurons. Among them, the effect of inhibiting the release of ampelopsin and camperol compared to myricetin was relatively high. The results are shown in FIG. 7.

When preparing the cosmetic composition containing the ampelopsin of the present invention, less than 0.5% (PEG400 5%, based on water 70%) may include ampelopsin, but the microneedle produced by the method of manufacturing the microneedle of the present invention The ampelopsin content can be increased to about 5% of the dry weight of the microneedle. Therefore, it is possible to manufacture a product containing a high concentration of polyphenols.

Claims (11)

A microneedle for improving skin wrinkles containing ampelopsin. delete delete According to claim 1, The microneedle microneedle for improving skin wrinkles, characterized in that it contains 0.01 to 10% by weight of ampelopsin relative to the total weight of the microneedle. The microneedle for improving skin wrinkles according to claim 1, wherein the material forming the microneedle is dissolved in the skin. The material of claim 4, wherein the material forming the microneedle is hyaluronic acid, sodium-carboxymethyl cellulose, sodium carboxymethyl cellulose (Na-CMC), vinylpyrrolidone-vinyl acetate copolymer, polyvinyl alcohol (Poly vinyl alcohol), polyvinyl pyrrolidone (Poly vinyl pyrrolidone), microneedle for improving skin wrinkles, characterized in that the saccharides or mixtures thereof. The microneedle for improving skin wrinkles according to claim 6, further comprising a plasticizer in addition to the material forming the microneedle. delete delete delete delete

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