KR102445155B1 - Soluble Microneedle for polyphenol delivery - Google Patents

Abstract

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

Description

Soluble Microneedle for polyphenol delivery

The present invention relates to a soluble microneedle, and to a soluble microneedle containing polyphenol having excellent anti-wrinkle effect. More specifically, the present invention provides a system for permeating the skin of polyphenols using the soluble microneedles.

Delivery of drugs through the skin is used in various fields and forms due to its ease of use. Drugs that pass through the skin are mainly for delivery to the systemic circulation through the skin, but in addition, drugs such as atopic therapeutics, whitening or wrinkle-improving cosmetics are used for the purpose of delivering to the organs of the skin itself. . Despite such convenience and functionality, there are many difficulties in delivering a 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 has a brick structure composed of keratin-rich keratinocytes, and a mortar filled with lipids such as ceramide, fatty acid, or wax between these keratinocytes. consists of structure. This structure acts as a barrier and has very low material permeability. Only low molecular weight components of 500 Da or less can be delivered into the skin by diffusion, and only substances with excellent lipid affinity can pass through the skin.

On the other hand, the discharge of neurotransmitters is caused by the fusion of a synaptic vesicle and a presynaptic membrane located at the nerve endings containing the neurotransmitter, and then a passage between the two boundaries is formed. The SNARE protein, a three-protein complex consisting of the vesicle protein VAMP (synaptobrevin), the plasma membrane binding protein Syntaxin 1a, and SNAP-25, provides the fundamental force for the fusion of the synaptic vesicle and the presynaptic membrane. Here, the opening of the neurotransmitter release pathway by membrane fusion between the synaptic vesicle and the presynaptic membrane is t-SNARE and vesicle ( It is a result according to the action of v-SNARE attached to the vesicle. When the membrane is fused, rearrangement of the lipid bilayer occurs. Since biological membranes strongly repel each other, these membranes do not spontaneously fuse, but a strong external force is applied to overcome the repulsive force between the membranes. It is known that the SNARE protein provides this force. As such, the formation of the SNARE complex is a key phenomenon of exocytosis, including the release of neurotransmitters.

Recently, efforts have been made to apply the plant polyphenols to situations where cosmetics or actual Botox cannot be used by taking advantage of the fact that they exert similar effects to Botox by controlling the release of neurotransmitters, but most polyphenols are Due to poor solubility, the amount that can be contained in the cosmetic formulation is limited, and thus the skin permeation amount is very low, so there is a problem that it is difficult to show efficacy.

Accordingly, an object of the present invention is to provide a soluble microneedle impregnated with a polyphenol component having an excellent skin permeability.

Polyphenol, which has excellent anti-wrinkle effect by blocking nerve transmission, is impregnated into the soluble microneedle, applied to the skin, and penetrates into the skin without pain by the microneedle, while the microneedle is dissolved by the moisture in the skin and the drug is delivered into the skin An object of the present invention is to provide a soluble microneedle that can be used.

An object of the present invention is to provide a skin delivery system that can be applied to the skin by solving the problem of limited use of polyphenols due to poor solubility in general cosmetic compositions, and improving the solubility of 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 polyphenol and having a wrinkle-improving effect. The material forming the microneedle may be dissolved or biodegradable in the skin, and when the microneedle is applied to the skin, the polyphenol is released by dissolving or disintegrating the microneedle, and the polyphenol can be stably delivered to the skin.

The present inventors have studied various administration systems, and as mentioned above in any system, polyphenol has a low solubility, so there is a limit to its application only by applying it to the skin by applying it to general cosmetics. After several efforts, the present inventors have made a surprising invention that can effectively reduce the transmission of neurotransmitters by containing polyphenols in soluble microneedles in the skin, thereby exhibiting a wrinkle-improving effect.

In order to achieve the above task, the microneedle must be soluble in the skin, and in order to form a soluble microneedle, hyaluronic acid, sodium-carboxymethyl cellulose (Na-CMC, Sodium carboxymethyl cellulose), vinyl skin water-soluble polymers such as rolidone-vinyl acetate copolymer, poly vinyl alcohol, and poly vinyl pyrrolidone; sugars such as xylose, sucrose, maltose, lactose, and trehalose; or mixtures thereof may be used. In particular, when the microneedle's skin penetration strength and dissolution rate in the skin are comprehensively considered, hyaluronic acid (Oligo-Hyaluronic acid), sodium-carboxymethyl cellulose (Na-CMC, Sodium carboxymethyl cellulose), and saccharides ) (more preferably, a mixture of trehalose) is more preferable, and even more preferably mixed with glycerine, which will be described later. Preferably, the microneedle according to the present invention may further include a plasticizer, a surfactant, a preservative, an anti-inflammatory agent, and the like, 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 alone. may be used as or in combination.

Preferably, the microneedle of the present invention may contain 0.01 to 10% by weight, more preferably 0.1 to 5% by weight, of polyphenol having a wrinkle-improving effect for controlling neurotransmitter release, based on the total weight of the microneedle. have.

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

Although the amount of the polyphenol component contained in the cosmetic composition was very small due to poor solubility, the microneedle of the present invention has an extremely low water content, so it can be applied to the skin in a high content without precipitation of the polyphenol component.

The polyphenol is kaempferol, quercetin, myricetin, luteolin, delcinidine, cyanidin, butane, ellagic acid, ampelopsin, hespiridine, aurantinidin (Aurantinidin), europinidin (Europinidin), pella Gonidin (Pelargonidin), malvidin (Malvidin), pheonidin (Pheonidin), petunidin (Petunidin), and may be any one or more selected from the group consisting of rosinidine (Rosinidin) and derivatives thereof, preferably may be any one or more selected from the group consisting of ampelopsin, hesperidin, or derivatives thereof.

In the present specification, the derivative of the polyphenol is a free form (free acid or base form) and all prodrugs, polymorphs, hydrates, solvates, tautomers, stereotypes Unless otherwise specified, such as isomers and the like, all pharmaceutically acceptable forms as described above are included, and all active forms of the compound are included. According to an embodiment of the present invention, the microneedle containing polyphenol inhibits the formation of a complex of SNARE to inhibit membrane fusion, and as a result inhibits the release of neurotransmitters, for example, acetylcholine, so that muscle cells contraction can be inhibited.

In addition, the present invention provides a microneedle patch system for administering (for delivery) polyphenol or a derivative thereof to which the above microneedles are attached.

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

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

In addition, the present invention provides a wrinkle-improving use of a microneedle containing polyphenol or a derivative thereof, which controls the neurotransmitter release and has a wrinkle-improving effect.

According to one embodiment of the present invention, the present invention provides a microneedle containing a polyphenol or a derivative thereof and inhibiting the formation of a SNARE complex.

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 permeation efficiency of polyphenol or a derivative thereof having a wrinkle-improving effect. The present invention also provides a method for skin administration of polyphenols or derivatives thereof, characterized in that such microneedles are used.

The following drawings attached to this specification illustrate preferred embodiments of the present invention, and serve to further understand the technical idea of the present invention together with the above-described content of the invention, so the present invention is limited to the matters described in those drawings It should not be construed as being limited.
1 is a view showing an example of several 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 is an exemplary view of polyphenols having a botox-like effect of the present invention. Myricetin, delcinidine, cyanidin, kaempferol, quercetin, fisetin, butane, luteolin, ellagic acid, EGCG (Epigallocatechin gallate), ampelopsin, hesperidin and derivatives thereof may be used.
Figure 4 shows the SNARE formation inhibition confirmation results of kaempferol, paroxetine, and ampelopsin. (Cell line: C2C12 (muscle cell) + NG108-15 (neuron 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 ampelopsin cream and ampelopsin microneedle on the human body.
7 shows the results of confirming the muscle contraction inhibitory effect of kaempferol, paroxetine, and ampelopsin.

Hereinafter, examples and the like will be described in detail to help the understanding of the present invention. However, the embodiments according to the present invention may be modified in various other 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 completely explain the present invention to those of ordinary skill in the art. The content described herein is expressed in wt% unless otherwise specified.

<Soluble Micro Needle Manufacturing>

Soluble Micro Needle is manufactured by the solution casting method, and the solution is cast into a mold, filled with a liquid in a micro mold with a vacuum or centrifuge, and then dried. .

<Manufacture of polyphenol microneedle that inhibits SNARE formation>

Microneedles having the composition shown in Table 1 below were prepared. Hyaluronic acid (Oligo-HA™), sodium carboxymethyl cellulose (Na-CMC) and Trehalose were dissolved in purified water and then Glycerin, PEG-40 hydrogenated castor oil (HCO-40™) and ampelopsin solution (ampelopsin 10%, DPG 90) %) to prepare an ampelopsin solution (DPG: Dipropylene glycol). After casting the prepared polyphenol dispersion solution to a silicone micro needle mold, centrifugation was performed at 3,000 rpm for 10 minutes to fill the micro mold.

After filling the solution, it was dried in a drying oven (70° C.) for 3 hours, and the Micro Needle was separated from the silicone mold using an adhesive film. The contents in Table 1 below are expressed in wt%.

The manufacturing process is shown in FIG. 1 .

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

<SnARE formation inhibition polyphenol oil-in-water cream production>

In order to compare the skin permeation amount of polyphenols impregnated in microneedle, as a comparative example, polyphenols were impregnated in a cream of a general emulsion in water formulation.

ingredient Formulation example C14-22 alcohol, C12-20 alkyl glucoside
(mixture C14-22 alcohol:C12-20 alkylglucoside = 80:20 weight ratio)
Glyceryl Stearate, PEG-100 Stearate
(Mixture 50:50 weight ratio)
Glyceryl Stearate
cetearyl alcohol
Polyglyceryl-3 methylglucose 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
Remaining amount (To 100)

<Drug release behavior>

Ampelopsin release from the microneedles prepared above was evaluated using a Franz diffusion cell equipped with pig skin (see FIG. 2 ). As an aqueous solution (acceptor solution), a PBS solution containing 30% by weight of DPG was used.

That is, using a Franz diffusion cell, the ampelopsin content of pig skin tissue and acceptor solution according to time was measured using Liquid Chromatography.

The amount of cream containing ampelopsin permeated through the skin was insignificant, but the ampelopsin impregnated in the micro needle directly penetrated into the skin by the needle and showed a higher amount of permeation than the cream.

The amount of the cream containing ampelopsin permeated through the skin was about 1 ug, which was insignificant, but the polyphenol impregnated in the microneedle directly penetrated into the skin by the needle and the permeation amount was 13 ug or more, which is about 13 times higher than the cream. The skin permeation amount was indicated. The results are shown in FIG. 5 .

<Wrinkle improvement effect>

Ampelopsin cream and microneedle impregnated with ampelopsin were treated daily for 12 weeks on the wrinkles around the eyes, and the degree of wrinkle improvement was confirmed through a silicone replica and wrinkle image analysis method. (N=20)

Compared to the ampelopsin cream, the microneedle impregnated with ampelopsin showed an improvement effect of more than 5 times, which confirmed that the ampelopsin effectively penetrated into the skin by the microneedle, and the wrinkle improvement effect was high. These results are shown in FIG. 6 .

<SNARE complex formation inhibition experiment>

To determine whether polyphenols such as ampelopsin, kaempferol, and myricetin inhibit the formation of the SNARE complex, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis was performed. It was confirmed whether 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 addition of a polyphenol compound at a concentration of 1 - 50 ppm. After each protein was dripped into a 1ml tube, a polyphenol compound was added, mixed, and reacted at room temperature for 30 minutes. After that, it was confirmed whether the SNARE complex was formed by electrophoresis on 12% SDS-PAGE.

It was confirmed that all of kaempferol, myricetin, and ampelopsin effectively inhibited the SNARE complex in a concentration-dependent manner. As a result of the experiment, it was confirmed that the ability to inhibit the formation of the SNARE complex was active at lower concentrations of kaempferol and ampelopsin compared to paroxetine. The SNARE formation inhibitory effect of paroxetine, ampelopsin (dihydroxymyricetin), kaempferol, and A440 was confirmed and shown in FIG. 4 . (Cell line: C2C12 (muscle cell) + NG108-15 (neuron cell) condition, concentration: ppm)

<Experiment to inhibit muscle contraction>

C2C12 cells were cultured in DMEM medium containing 10% fetal calf serum and 1% antibiotics on the plate. Then, neuroblasts were additionally co-cultured on the same plate. Then, when the cell contraction of the C2C12 cells starts, the number of contractions of the C2C12 cells is measured for 30 seconds, and after removing all the medium, it is washed 3 times with PBS. made it After that, the number of contractions of C2C12 cells was measured again for 30 seconds to confirm the degree of muscle contraction inhibition.

It was confirmed that kaempferol, myricetin, and ampelopsin reduced the number of contractions of C2C12 cells by inhibiting the release of neurotransmitters from neurons. Among them, ampelopsin and kaempferol were measured to have a relatively high effect on inhibiting release compared to myricetin. The results are shown in FIG. 7 .

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

Claims (5)

As a water-soluble microneedle containing any one or more polyphenols selected from the group consisting of ampelopsin, kaempferol, quercetin, myricetin and luteolin,
The microneedle does not contain liposomes, the polyphenol is impregnated in the structure of the microneedle, and the material forming the structure of the water-soluble microneedle contains a water-soluble polymer,
The water-soluble microneedle, characterized in that when the water-soluble microneedle is applied to the skin, the polyphenol is released by dissolving or disintegrating the microneedle by moisture in the skin. The water-soluble microneedle according to claim 1, wherein the water-soluble microneedle contains 0.01 to 10% by weight of polyphenol based on the total weight of the microneedle. According to claim 1, wherein the material forming the structure of the water-soluble microneedle is hyaluronic acid (Hyaluronic acid), sodium-carboxymethyl cellulose (Sodium carboxymethyl cellulose), vinyl pyrrolidone-vinyl acetate copolymer, polyvinyl alcohol (Poly vinyl alcohol), polyvinyl pyrrolidone (Poly vinyl pyrrolidone), a water-soluble microneedle, characterized in that the saccharide or a mixture thereof. The water-soluble microneedle according to claim 3, wherein the water-soluble microneedle further comprises a plasticizer in addition to the material forming the structure of the microneedle. A microneedle patch for wrinkle improvement, comprising the water-soluble microneedle according to any one of claims 1 to 4,
The polyphenol is a microneedle patch for wrinkle improvement, characterized in that ampelopsin.

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