KR101983570B1 - Lipid nano particle composition for improvement of orobol stability and method for production of it - Google Patents

Abstract

The present invention relates to a lipid nanoparticle composition comprising, as active ingredients, orobol, solid and liquid lipids, and a surfactant, and a method for producing the same. According to the present invention, it has been confirmed that stability of orobol is improved as compared with conventional emulsion formulations. Also, it has been confirmed that the lipid nanoparticles according to the present invention do not cause human skin irritation, and that the skin residual amount of orobol can be also increased. Thus, the lipid nanoparticle composition comprising orobol according to the present invention, when applied to the skin, allows better topical dermal delivery of orobol without causing skin irritation, and thus is expected to be an effective cosmetic formulation or composition with improved orobol stability.

Description

TECHNICAL FIELD The present invention relates to a lipid nanoparticle composition for improving the stability of an oval ball,

The present invention relates to a nanoparticle for improving the stability of a bioabsorbable material and a method for producing the nanoparticle.

Soy isoflavones have attracted attention as new materials for the treatment of cancer, obesity and cardiovascular disease due to antioxidant and anti-cancer effects. Recently, it has been shown that it has an important effect in skin diseases. Genistein and daidzein, which are isoflavones, have been shown to inhibit UV-induced oxidation when applied topically (H Wei et al. / Cancer Letters 185.1 (2002) 21-29). This study suggests that isoflavones may be used as a treatment for skin wrinkles and skin cancer. In particular, the metabolism of genistein, Orobol (5,7,3,4, -tetrahydroxyisoflavone), is the most efficacious of the isoflavone family of skin (Seoul National University Industry Collaboration Foundation, Bob Sno / ≪ / RTI > 1016026290000).

Although Orobolt was not found in nature, Orogenol has recently been attracted attention as a next-generation skin treatment material by succeeding in mass production by attaching OH group to genistein (catechol type structure material manufactured by Seoul National University Industry Collaboration Team / tyrosinase, Method and its application, 1015666720000). However, the isoflavones genistein and daidzein react with organic solvents when exposed to sunlight, resulting in degradation (MM Kelly et al. / Environmental science & technology 46.10 (2012) 5396-5403) And the browning caused by this occurs. Therefore, formulation studies that can maintain stability for skin application are essential, but there are few studies on the formulation of ooloball.

On the other hand, examples of nanoparticles that can improve the stability of the active ingredient include lipid nanoparticle formulations. In particular, Nanostructured Lipid Carrier (NLC), which has been studied since the early 2000s, is an improved formulation of SLN (Solid Lipid Nanoparticle) developed in the 1990s.

SLN is a formulation in which a solid lipid forms a hydrophobic core unlike emulsion or liposome. NLC is a formulation made by mixing solid lipid and liquid lipid to improve SLN. Both SLN and NLC lipid nanoparticles maintain the solids content at room temperature, which enhances the stability of the efficacious components trapped inside and facilitates release behavior compared to other formulations. It is also safe to use on the skin because it can be composed of biodegradable materials. Indeed, studies have shown that when tretinoin is sutured to SLN and NLC, it improves light stability and maximizes efficacy (K. Raza et al. / International journal of pharmaceutics 456 (2013) 6572). Also, there is a study showing that SLN formulation improved retinol stability rather than simple emulsion when retinol was stored for 6 months (R. H. Muet al. / Advanced Drug Delivery Reviews 59 (2007) 522530).

Therefore, in the present invention, it is intended to develop a formulation and a composition capable of enhancing the stability of the oolor ball by applying lipid nanoparticles to the oolol ball, which is a poorly stable active ingredient. The content of the oolol ball, , And to evaluate the improved stability by observing the color change.

Korean Patent Registration No. 1016026290000. Korean Patent Registration No. 1015666720000.

The present invention provides a method for producing lipid nanoparticles and an emulsion produced thereby by enclosing a poorly stable oolium into a solid lipid or a mixture of 'solid and liquid lipid' to improve the stability of the oolab will be.

The present invention relates to a process for preparing a liquid product comprising the steps of: (a) preparing an oil phase by adding and dissolving Orobol, solid lipid and liquid lipid in a solvent; (B) a step of adding a surfactant-added aqueous phase to the oil phase prepared in the step (a) and homogeneously mixing to form nanoparticles; And (c) cooling the nanoparticles to a temperature below the melting point of the solid lipid to solidify the solid lipid present in the core of the nanoparticles. And a manufacturing method thereof.

In the process for producing the lipid-containing lipid nanoparticles of the present invention, it is preferable that the step (a) further comprises adding a liquid lipid to the solvent.

In the method for producing the lipid-containing lipid nanoparticles according to the present invention, the solid lipid may include, for example, cetyl palmitate, Precirol ATO 5, Compritol 888 ATO, glyceryl palmitostearate, glyceryl monostearate, glyceryl behenate, glyceryl palmitostearate, shea butter, and the like. but are not limited to, butter, Cocoa butter, stearic acid, palmitic acid, decanoic acid, stearyl alcohol, palmityl alcohol and myristyl alcohol myristyl alcohol, and the like.

In the method of manufacturing the lipid-containing lipid nanoparticles of the present invention, the liquid lipid may be, for example, Miglyol, Kernel Oil, Seed oil, flower oil, fruit oil, Capmul MCM, olive oil, eutanol G, cetiol CC, medium-chain triglyceride, squalane, coconut oil, jojoba oil , Almond oil, olive oil, argan oil, foolish rice oil, and camellia oil.

In the production method of the lipid nanoparticles containing the Ouroborate of the present invention, the solvent may be, for example, transcutol, ethyl acetate, ether, n-hexane, cyclohexane, tetrahydrofuran, methylene, chloride, formamide, One or more members selected from the group consisting of dimethylsulfoxide, isopropanol, methanol, ethanol, chloroform, propanol, 1,3-butylene glycol, propylene glycol, glycerin, 1,2-pentanediol, di-panthenol, dipropylene glycol and acetone .

In the production method of the lipid nanoparticles containing the Ouro Ball according to the present invention, the surfactant may be an alkyl ether sulfate such as Brij, Poloxamer, Tween, Span, And may be one or more selected from the group consisting of alcohol ethoxylate, low foaming surfactant, EO-PO block polymer, polyoxyethylene sorbitan fatty acid ester, and PEG .

In the production method of the lipid-containing lipid nanoparticles of the present invention, the steps (a) and (b) are preferably carried out at 30 to 90 ° C.

In the method for producing the lipid-containing lipid nanoparticles of the present invention, the water phase of step (b) may be an aqueous buffer solution having a pH of 4.0 to 8.5 which does not cause water or skin irritation.

In the method for preparing the lipid nanoparticles containing the oolabic of the present invention, the nanoparticles of the step (b) may be formed by, for example, a bath-sonicator, a tip-sonicator, , A high-pressure emulsifier (Microfluidzer), and an SPG membrane.

In the method for producing the lipid-containing lipid nanoparticles of the present invention, the cooling in step (c) is preferably performed at room temperature.

The present invention provides an emulsion comprising an emulsion containing a cured solid lipid, wherein the cured solid lipid comprises a spheroid.

In the emulsion of the present invention, the emulsion may be one in which the liquid lipid forms lipid nanoparticles and the solid lipid cured in the lipid nanoparticles is dispersed.

In the emulsion of the present invention, the solid lipid may be, for example, Cetyl palmitate, Precirol ATO 5, Compritol 888 ATO, trimyristin, tripalmitin ), Tristearin, glyceryl monostearate, glyceryl behenate, glyceryl palmitostearate, Shea butter, Cocoa butter ), Stearic acid, palmitic acid, decanoic acid, stearyl alcohol, palmityl alcohol, and myristyl alcohol. It can be one or more selected.

In the emulsion of the present invention, the liquid lipid may be, for example, Miglyol, Kernel Oil, Seed oil, flower oil, fruit oil, Capmul MCM, olive oil, eutanol G, cetiol CC, medium-chain triglyceride, squalane, coconut oil, jojoba oil, almond oil, olive oil, Argan oil, foolish rice oil, and camellia oil.

In the emulsion of the present invention, the emulsion is preferably a surfactant, selected from the group consisting of Alkyl Ether Sulfate such as Brij, Poloxamer, Tween, Span, A polyoxyethylene sorbitan fatty acid ester, and PEG may be used as the surfactant in the present invention. The surfactant may be one selected from the group consisting of Alcohol Ethoxylate, Low Foaming Surfactant, EO-PO Block Polymer, polyoxyethylene sorbitan fatty acid ester and PEG.

The present invention provides a cosmetic composition containing the emulsion of the present invention.

 The composition according to the present invention is a formulation for stable skin delivery of the oolop component, which is an effective ingredient of the anti-aging effect of skin, which has not been sufficiently studied in the formulation until now, and overcomes the stability problem that the conventional oolol composition can not solve, It is expected to be a formulation.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view of a process for preparing lipid nanoparticles containing oolobium.
Figure 2 shows the formation of precipitates in the formulations after preparation of the lipid nanoparticles of Comparative Examples 1 to 6.
FIG. 3 is a graph showing changes in the content of the oololbu after 4 weeks of the composition of Comparative Example 7 and Examples 2, 7 and 8.
4 is a graph showing a change in the content of the oololbu after 4 weeks' standing of the composition of Comparative Example 7 and Examples 5, 9 and 10.
FIG. 5 is a photograph showing the color change of the formulation immediately after the preparation of the compositions of Comparative Examples 7 to 8 and Examples 2 and 5 and after leaving for one week.
Fig. 6 shows the skin residual amount of the composition of Comparative Example 7 and Examples 2 and 5.
Fig. 7 shows the particle size distribution of the lipid nanoparticles of Example 5. Fig.
Fig. 8 shows the morphological shape of the lipid nanoparticles of Example 5. Fig.

The present invention provides an emulsion comprising an emulsion containing a cured solid lipid, wherein the cured solid lipid is loaded with a spherical ball, and a cosmetic composition containing the emulsion.

On the other hand, the present invention relates to a process for producing an oil phase comprising the steps of: (a) preparing an oil phase by adding and dissolving Orobol, solid lipid in a solvent; (B) a step of adding a surfactant-added aqueous phase to the oil phase prepared in the step (a) and homogeneously mixing to form nanoparticles; (C) cooling the nanoparticles to a temperature below the melting point of the solid lipid to solidify the solid lipid present in the core of the formed nanoparticles.

Hereinafter, the process for preparing the lipid nanoparticles of the present invention will be described in detail for each step.

<Step (a): Orobor  And dissolving the solid lipid in ethanol to prepare an oil phase.

Orobol and solid lipids are added to ethanol and heated to completely dissolve. At this time, the liquid lipid may be further added to ethanol. The order of addition may vary depending on the lipid, and the temperature may be raised from about 40 ° C to 80 ° C. Preferably, the temperature at which the solid lipid can be completely dissolved (the melting point of the solid lipid + 10 ° C) may be suitable. More preferably, it is preferable to use, as solid lipids, waxes which can be melted even when heated to a high temperature in consideration of thermal stability. Further, stirring may be carried out for complete dissolution.

&Lt; Step (b): After mixing the oil phase and water phase prepared above, Granulated  Step>

The oil phase prepared above and the water phase prepared separately are homogeneously mixed. When mixing, the mixture may be stirred using a magnetic bar or a stirrer, or may be stirred using a homogenizer. Preferably at the same temperature as the oil phase at the time of stirring. As the water phase, distilled water or a pH buffer may be used, and one or more types of stabilizers or surfactants may be included. The temperature of the aqueous phase may be from about 40 캜 to 80 캜, preferably a temperature such as the above-mentioned oil phase may be suitable.

Sufficient energy is applied to the mixture of the oil phase and water phase to form nanoparticles. For making the mixture solution into nanoparticles, a bath-sonicator, a tip-sonicator, a high-pressure emulsifier (Microfluidzer), an SPG membrane and the like can be used, Lt; / RTI &gt;

&Lt; Step (c): Cooling the Formulation >

By cooling the formulation, the solid lipid present in the core of the nanoparticles formed above is solidified to ultimately form lipid nanoparticles. The formulation can be allowed to stand for a short time in a cold state to be hardened. Preferably, the formulation is allowed to stand at room temperature for a long time and hardened.

A schematic diagram of the entire production process described above is shown in Fig. The lipid nanoparticles of the present invention prepared in the above process are present in the o / w emulsion. At this time, the lipid nanoparticles may be prepared using only 'solid lipids' or may be produced using 'solid lipids and liquid lipids'. In the present invention, the solid lipid is hardened through the process of the step (c), and the oval ball is mounted inside the solid lipid. In addition, when the lipid nanoparticles are produced by using the 'solid lipid and liquid lipid', the hardened solid lipid is dispersed in the lipid lipid, and the lipid is in the solid lipid.

[ Example  1 ~ 6: Depending on the type and amount of solid lipids Orobor  Preparation of lipid-containing lipid nanoparticles]

Orobol and solid lipids or liquid lipids were added to ethanol and completely dissolved by warming. The order of addition may vary depending on the lipids, and the temperature was raised to about 40-80 ° C according to the lipid selected below. The temperature at which the solid lipid is completely melted is suitable. It is preferable to heat the solid lipid to the melting point + 10 ° C. Further, stirring may be carried out in order to dissolve completely.

Meanwhile, the oil phase prepared above and the water phase prepared separately were homogeneously mixed. At the time of mixing, it is preferable to carry out stirring using a magnetic bar or a stirrer, or agitation using homogenizer or agar. In addition, it is preferable to warm to the same temperature as the oil phase at the time of stirring. As the water phase, distilled water or a pH buffer may be used, and one or more types of stabilizers or surfactants may be included. The temperature of the water phase may be warmed to about 40 to 80 DEG C, preferably a temperature such as the above-mentioned oil phase is suitable.

On the other hand, sufficient energy was applied to the mixed liquid of the oil phase and water phase to form nanoparticles. For the nanoparticle mixture, either a bath-sonicator, a tip-sonicator, a high-pressure emulsifier (Microfluidizer), or an SPG membrane was selectively used. At this time, it is preferable to perform heating by heating to the same temperature as the oil phase.

Meanwhile, By cooling the formulation prepared above, the solid lipid present in the core of the nanoparticles formed above was hardened to form the final lipid nanoparticles. The formulation can be allowed to stand for a short time in a cold state to be hardened. Preferably, the formulation is allowed to stand at room temperature for a long time and hardened.

Table 1 below shows the composition components of Examples 1 to 6 prepared by the above production method and their addition amounts.

The composition components and the addition amounts of Examples 1 to 3 Classification ingredient Example 1 Example 2 Example 3 Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Active ingredient Orobol 5 1.1 5 1.0 5 0.8 Solid lipid Cocoa butter 50 11.0 100 19.8 200 33.1 Liquid lipid Capmul MCM - - - - - - Surfactants Tween 20 200 44.0 200 39.6 200 33.1 Dissolution aid Transcutol 200 44.0 200 39.6 200 33.1

The composition components and the addition amounts of Examples 4 to 6 Classification ingredient Example 4 Example 5 Example 6 Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Active ingredient Orobol 5 1.1 5 1.0 5 0.8 Solid lipid Shea butter 50 11.0 100 19.8 200 33.1 Liquid lipid Capmul MCM - - - - - - Surfactants Tween 20 200 44.0 200 39.6 200 33.1 Dissolution aid Transcutol 200 44.0 200 39.6 200 33.1

[ Experimental Example  1: Example  1 to 6 Orobor  The average particle size of the lipid-containing lipid nanoparticles, Ourob  Measurement of the content of the filler and the content in the formulation]

Examples 1 to 6 An ELS spectrophotometer (ELS 8000; Otsuka Electronics Co., Ltd., Tokyo, Japan) was used to measure the average size of lipid nanoparticles. The lipid nanoparticles were transferred to a quartz cell and then repeatedly measured at 25 ° C three times. To obtain only the lipid nanoparticles encapsulated with oval balls, the formulation was appropriately diluted and then placed in an amicon tube having a certain pore size and centrifuged for 20 minutes. Thereafter, the orobol solution, which is not encapsulated in the nanoparticles but dissolved in the water and exited from the tube, is appropriately diluted and injected into high-performance liquid chromatography (Waters; Waters Co., Milford, Mass., USA) The amount of the oolball was quantified.

The rate of inclusion of the Orobolt is 100% of the total, minus the percentage of Orolobals not enclosed. In quantitative analysis, the composition of the mobile phase was determined to be 10 mM Na ₂ H ₂ PO ₄ (pH = 6): ACN = 40:60 and the flow rate was 1.0 mL / min. The column used was a Kinetex C18 column (4.6 x 250 mm, 5 μm; Phenomenex Inc., Torrance, CA, USA) and the volume of the sample injected was 20 μL. The detector used was a UV detector, and the detection wavelength was 245 nm.

Examples 1-3 The average particle size of the lipid nanoparticles and the inclusion rate and content of the oolubol parameter Example 1 Example 2 Example 3 Particle mean size (nm) 115 ± 0.42 113 ± 1.27 156 ± 1.10 Orobol inclusion rate (%) 95.8 ± 0.31 95.7 ± 0.16 92.9 ± 0.13 Orobol content (%) 1.05 ± 0.002 0.95 ± 0.001 0.77 ± 0.001

Examples 4 to 6 Average particle size of lipid nanoparticles and the inclusion rate and content of the oolubol parameter Example 4 Example 5 Example 6 Particle mean size (nm) 129 ± 0.94 106 ± 1.72 131 ± 1.43 Orobol inclusion rate (%) 92.5 ± 0.10 95.5 ± 0.25 95.7 ± 0.12 Orobol content (%) 1.02 0.001 0.94 ± 0.002 0.79 ± 0.001

From the results of Tables 3 and 4, it can be seen that the average particle size is about 110 to 160 nm and the average particle size is increased when the amount of solid lipid is 200 mg. On the other hand, when the amount of solid lipid was 100 mg, the smallest size was obtained.

In addition, the inclusion rate of oolubol was high regardless of the solid lipid, and the formulations containing 100 mg of solid lipid showed high inclusion rate and content. Therefore, it was concluded that the size, encapsulation rate and content of lipid nanoparticles were influenced by the amount of solid lipid. Based on this, it was found that the setting of the amount of solid lipid is important.

[ Comparative Example  1 ~ 6: No addition of surfactant Orobor  Preparation of lipid-containing lipid nanoparticles]

Oolvol-containing lipid nanoparticles were prepared according to the methods described in Examples 1 to 6, except that no surfactant was added.

The composition components and the addition amounts of Comparative Examples 1 to 3 Classification ingredient Comparative Example 1 Comparative Example 2 Comparative Example 3 Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Active ingredient Orobol 5 2.0 5 1.6 5 1.2 Solid lipid Cocoa butter 50 19.6 100 32.8 200 49.4 Liquid lipid Capmul MCM - - - - - - Surfactants Tween 20 - - - - - - Dissolution aid Transcutol 200 78.4 200 65.6 200 49.4

The composition components and the addition amounts of Comparative Examples 4 to 6 Classification ingredient Comparative Example 4 Comparative Example 5 Comparative Example 6 Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Active ingredient Orobol 5 2.0 5 1.6 5 1.2 Solid lipid Shea butter 50 19.6 100 32.8 200 49.4 Liquid lipid Capmul MCM - - - - - - Surfactants Tween 20 - - - - - - Dissolution aid Transcutol 200 78.4 200 65.6 200 49.4

[ Experimental Example  2: Comparative Example  1 to 6 Orobor  Observation of formation of precipitate after preparation of lipid-containing nanoparticles]

After preparation of the lipid nanoparticles of Comparative Examples 1 to 6, it was observed whether a precipitate was formed after a certain period of time to evaluate whether or not the lipid nanoparticles were stably dispersed in the aqueous phase. After preparation of the lipid nanoparticles, the photograph of the formulation is shown in Fig. As a result, in Comparative Examples 1 to 6, in which no surfactant was present, lipid nanoparticles were not dispersed evenly, and aggregation and precipitation were observed. As a result, it has been found that at least one kind of surfactant is essential for obtaining a formulation in which the lipid nanoparticles containing the oolone ball are stable and uniformly dispersed in the aqueous phase.

[ Example  7 to 10: Addition of liquid lipid Orobor  Preparation of lipid-containing lipid nanoparticles]

The oval ball-containing lipid nanoparticles were prepared according to the methods described in Examples 1 to 6, except that the amount of liquid lipid was varied to prepare the oval ball-containing lipid nanoparticles.

The composition components and the addition amounts of Examples 2, 7, and 8 Classification ingredient Example 2 Example 7 Example 8 Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Active ingredient Orobol 5 1.0 5 0.9 5 0.9 Solid lipid Cocoa butter 100 19.8 100 18.7 100 18.0 Liquid lipid Capmul MCM 0 0 30 5.6 50 9.0 Surfactants Tween 20 200 39.6 200 37.4 200 36.0 Dissolution aid Transcutol 200 39.6 200 37.4 200 36.0

The composition components and the addition amounts of Examples 5, 9 and 10 Classification ingredient Example 5 Example 9 Example 10 Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Active ingredient Orobol 5 1.0 5 0.9 5 0.9 Solid lipid Shea butter 100 19.8 100 18.7 100 18.0 Liquid lipid Capmul MCM 0 0 30 5.6 50 9.0 Surfactants Tween 20 200 39.6 200 37.4 200 36.0 Dissolution aid Transcutol 200 39.6 200 37.4 200 36.0

[ Experimental Example  3: Example  2, 5, and 7 to 10 Orobor  The average particle size of the lipid-containing lipid nanoparticles, the loading rate of the oolubol, and the content in the formulation]

The average particle size of the lipid nanoparticles, the loading rate and content of the oval balls in Examples 2, 5 and 7 to 10 were measured in the same manner as in Experimental Example 1.

The average particle size of the lipid nanoparticles of Examples 2, 7 and 8 and the content and content of the oolabol parameter Example 2 Example 7 Example 8 Particle mean size (nm) 118 ± 1.40 450 ± 16.3 743 ± 3.80 Orobol inclusion rate (%) 98.6 ± 0.01 98.4 ± 0.002 98.1 ± 0.01 Orobol content (%) 0.98 ± 0.001 0.92 ± 0.001 0.88 ± 0.001

Examples 5, 9 and 10 The average particle size of lipid nanoparticles and the inclusion rate and content of the oolabol parameter Example 5 Example 9 Example 10 Particle mean size (nm) 165 ± 2.25 289 ± 5.19 324 ± 9.99 Orobol inclusion rate (%) 98.2 ± 0.01 98.1 ± 0.01 98.3 ± 0.01 Orobol content (%) 0.97 ± 0.002 0.91 ± 0.001 0.86 ± 0.001

The results showed that the average particle size of lipid nanoparticles increased with increasing amount of liquid lipid. This tendency varied depending on the kinds of solid lipids, and in the case of Examples 5, 9 and 10 in which the solid lipid was shea butter, the average particle size was maintained at about 330 nm or less. However, the inclusion rate of Orobolt was higher than 98% regardless of addition of liquid lipid.

However, the addition or the amount of liquid lipid will be important as the amount of liquid lipid is excessively large, thereby increasing the average particle size of the formulation and reducing the oval content in the formulation.

[ Comparative Example  7 ~ 8: Without using solid lipids Orobor  contain emulsion  And ethanol solution]

As a control, the composition of the following Table 11 was prepared to prepare the oolovol containing emulsion and the ethanol solution without using solid lipids, and then mixed using the method of Example 1 above.

Composition of emulsion containing ethanol and ethanol solution Classification ingredient Comparative Example 7 Comparative Example 8 Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Active ingredient Orobol 5 0.05 5 0.05 Liquid lipid Capmul MCM 2 mL 19.99 - - Surfactants Labrasol 1.43 mL 14.30 - - Dissolution aid Transcutol 2.87 mL 28.68 - - Aqueous solvent DW 3.7 mL 36.98 - - Organic solvent Ethanol - - 10 mL 99.95

[ Experimental Example  4: Comparative Example  7 Orobor  contain Emulsion and Example  2, 5, and 7 to 10 Orobor  Evaluation of the stability of lipid nanoparticles containing

As a stability evaluation, the average particle size was measured and the content of ovalbone relative to the initial content was measured immediately after production. Each sample was allowed to stand at room temperature and light for 4 weeks, and the change of the average particle size and the content of the ool ball was evaluated over time. The average particle size of the formulation and the content of the oolubol were measured in the same manner as in Experimental Example 1.

The average particle size (nm) change of the composition of Comparative Example 7 and Examples 2, 5 and 7 to 10 for 4 weeks Comparative Example 7 Example 2 Example 5 Example 7 Example 8 Example 9 Example 10 Immediately after manufacture (Day 0) 252 ± 23 118 ± 1 165 ± 2 450 ± 16 743 ± 4 289 ± 5 324 ± 10 Day 3 263 ± 21 122 ± 2 164 ± 4 455 ± 34 502 ± 24 246 ± 8 288 ± 4 Day 5 224 ± 9 121 ± 1 170 ± 1 456 ± 35 741 ± 34 301 ± 10 325 ± 3 Day 7 217 ± 42 110 ± 2 160 ± 2 416 ± 4 563 ± 10 298 ± 7 323 ± 1 Day 14 217 ± 52 111 ± 2 160 ± 3 416 ± 4 563 ± 12 298 ± 9 323 ± 1 Day 21 344 ± 14 106 ± 1 151 ± 1 375 ± 1 490 ± 9 265 ± 2 330 ± 2 Day 28 391 ± 27 106 ± 1 154 ± 1 370 ± 5 798 ± 38 271 ± 2 348 ± 7

(%) Change of the ovalboy content of the composition of Comparative Example 7 and Examples 2, 5 and 7 to 10 for 4 weeks Comparative Example 7 Example 2 Example 5 Example 7 Example 8 Example 9 Example 10 Immediately after manufacture (Day 0) 100 100 100 100 100 100 100 Day 3 87.2 ± 2.8 99.7 ± 0.0 100 ± 0 99.8 ± 0.1 98.0 ± 0.1 99.9 ± 0.0 99.2 0.0 Day 5 76.9 ± 2.2 99.3 ± 0.0 99.7 ± 0.0 98.6 ± 0.0 98.5 ± 0.0 98.2 ± 0.1 96.5 ± 0.1 Day 7 55.1 ± 0.6 97.0 ± 0.7 95.6 ± 1.6 100 ± 0 94.7 ± 1.8 97.6 ± 2.1 93.2 ± 0.6 Day 14 40.9 ± 2.2 85.5 ± 0.5 87.8 ± 0.0 91.2 ± 0.0 86.8 ± 0.0 92.9 ± 0.0 88.7 ± 0.1 Day 21 39.1 ± 1.5 82.1 ± 0.1 78.3 ± 1.5 75.8 ± 1.4 78.7 ± 1.4 76.7 ± 0.0 80.3 ± 2.7 Day 28 27.9 ± 0.4 75.1 ± 3.1 74.5 ± 0.1 77.9 ± 1.8 70.2 ± 0.7 80.2 ± 0.2 80.0 ± 0.5

The results of Table 12 show that the average particle size after 4 weeks of standing did not change much in both the Comparative Example 7 (emulsion) composition and the examples (lipid nanoparticles). Comparative Example 7 showed a tendency to increase the particle size due to slight aggregation over time but did not settle for 4 weeks. In contrast, the above examples did not change particle size significantly over time except for Example 8, and no precipitate was formed for 4 weeks.

On the other hand, according to the results shown in Table 13 and Figs. 3 and 4, the content of oolabol in all the compositions continued to decrease with time as compared with the initial content immediately after preparation. This is probably due to instability due to temperature or light of the material itself. However, such a reduction in the content of the lipid nanoparticle composition was much more relaxed in the examples of the lipid nanoparticle composition.

The content of oolabol in the composition of Comparative Example 7, which is a typical emulsion formulation, dropped to about 28% over 4 weeks, while the composition of the lipid nanoparticle formulations was maintained at about 75% or more except Example 8 It was confirmed that the lipid nanoparticle composition increased the long term content stability of the Orobolt by about 2.5 times as compared with the general emulsion.

The improvement in the stability of such lipid nanoparticle formulations is due to the fact that the core of the formulation is a solid and the phenomenon that the enclosed oolium interacts with or is released from the outer water phase is relatively less than that of a typical emulsion formulation in which the core is a liquid As shown in Fig. All of the lipid nanoparticle compositions retained similar contents, among which the average particle sizes of Examples 2 and 5 were the smallest, and the ratio of solid lipids in the nanoparticle cores was the highest. The compositions of Examples 2 and 5 are considered as preferred compositions.

[ Experimental Example  5: Comparative Example  7 to 8 and Example  2 and 5 Composition Observation of Formulation Color Change Immediately After and After 2 Weeks]

On the basis of the phenomenon that the color of the solution changed to yellow when the oolong ball was dissolved in the aqueous phase and became unstable, in order to examine the stability improvement of the oololbuhn, Comparative Examples 7 to 8 and Examples 2 and 5 The color change of the formulation was observed after standing. A color change photograph of the formulation is shown in Fig.

As a result, the composition of Comparative Example 7 was slightly yellow even after the preparation, and after standing for 2 weeks, it was confirmed that the composition had a darker yellow color. That is, in a typical emulsion formulation, it is considered that the color of the formulation changes and the stability of the oval ball is not maintained because there is interaction between the outer aqueous phase and the enclosed ourobium. However, in the case of Examples 2 and 5, which were the lipid nanoparticle compositions of the present invention, they were the same as the color of the formulation immediately after preparation even after 2 weeks. Therefore, in the case of the above-mentioned lipid nanoparticle composition in which the core of the nanoparticle is solid, it is confirmed that the enclosed oolob is less likely to interact with the outer oolong, and that the stability of the content of oolubol can be maintained without changing the color of the formulation will be.

[ Comparative Example  9 to 10: For skin irritation test, Oroball  Preparation of Lipid Nanoparticles Not Contained]

Using the method described in Example 1 above, lipid nanoparticles free from ovalballs were prepared from the composition components and contents shown in Tables 14 and 15 below.

Production of lipid nanoparticles without oval balls for skin irritation test Classification ingredient Comparative Example 9 Example 7 Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Active ingredient Orobol - - 5 0.9 Solid lipid Cocoa butter 100 18.9 100 18.7 Liquid lipid Capmul MCM 30 5.7 30 5.6 Surfactants Tween 20 200 37.7 200 37.4 Dissolution aid Transcutol 200 37.7 200 37.4

Production of lipid nanoparticles without oval balls for skin irritation test Classification ingredient Comparative Example 10 Example 9 Amount (mg) Content ratio (%) Amount (mg) Content ratio (%) Active ingredient Orobol - - 5 0.9 Solid lipid Shea butter 100 18.9 100 18.7 Liquid lipid Capmul MCM 30 5.7 30 5.6 Surfactants Tween 20 200 37.7 200 37.4 Dissolution aid Transcutol 200 37.7 200 37.4

[ Experimental Example  6: Comparative Example  9 to 10 Example  7 and &lt; RTI ID = 0.0 &gt; 9 &lt; / RTI &

When the compositions of Comparative Examples 9 to 10 containing no active ingredient oolovol and the compositions of Examples 7 and 9 containing oolol were applied in a single round and cumulatively, the irritation to human skin was evaluated. A total of 30 subjects were selected to meet the criteria. The skin test was applied to the subject 's back for 24 hours. The cumulative test was applied to the skin for 5 weeks for 2 weeks. After 2 weeks of rest, the skin was stimulated for 48 hours. Skin sensitization responses were scored according to the International Society of Contact Dermatology and the American Society of Cosmetics Safety Assessment Guidelines.

The single and cumulative skin irritation scores of the compositions of Comparative Examples 9 to 10 and Examples 7 and 9, parameter Comparative Example 9 Comparative Example 10 Example 7 Example 9 Skin irritation
score Single application 0.017 (non-magnetic) 0.017 (non-magnetic) 0.017 (non-magnetic) 0.017 (non-magnetic) Cumulative application 0.00 (non-magnetic) 0.00 (non-magnetic) 0.02 (non-magnetic) 0.01 (non-magnetic)

As a result, all of the lipid nanoparticle compositions were found not to be irritating to skin, regardless of the content of the active ingredient, ourobil. In other words, it was confirmed that the active ingredient, Orobol, and all the components used were biocompatible. Based on this, it is thought that the lipid nanoparticle composition can be a nano dosage form capable of improving the stability of the ovalbone, and at the same time, be a suitable cosmetic formulation having no irritation to human skin.

[ Experimental Example  7: Comparative Example  7 and Example  2 and 5 composition Ourob  Skin residual amount measurement]

The Keshary-Chien diffusion cell was used for measuring the skin residual amount of the oolium according to Comparative Example 7 and Example 2 and 5 lipid nanoparticles in the emulsion formulations. In addition, Strat-M ^® membrane (diameter 25 mm; Merck Millipore Ltd., Burlington, MA, USA) was used as an artificial membrane to represent the skin. The Strat-M ^® membrane was placed between the donor and receptor cells with the smooth side facing the donor cell and clamped using a clamp. The receptor cells were filled with PBS with 0.5% tween 80 solution and stirred at 600 rpm.

After the donor cells into about 1 mL of the embodiment, after sealing was 6 hours and the amount of ORO ball remaining in Strat-M ^® membrane was measured by high performance liquid chromatography. The total diffusing cell was tested at 32 ° C and the residual measurement was repeated at least three times. The skin residual amount results are shown in Fig.

The results showed that the residual amount of the oolab was about 3.5 times higher than that of Comparative Example 7, which is a typical emulsion formulation, after application of the lipid nanoparticle compositions of Examples 2 and 5. As a result, the lipid nanoparticle composition can be expected to improve the stability of the ourobule, increase the functional effect due to the skin irritation, and increase the skin residual amount.

[ Experimental Example  8: Orobor  Measurement of particle size distribution of lipid nanoparticles containing [

Example 5 The particle size distribution of lipid nanoparticles was measured in the same manner as in Experimental Example 1 using an ELS spectrophotometer. The particle size distribution graph is shown in FIG.

As a result, it was confirmed that the prepared lipid nanoparticles had a uniform particle size distribution in a normal distribution curve shape having a range of about 150 nm.

[ Experimental Example  9: Orobor  Observation of morphological shape of lipid nanoparticles containing [

Example 5 The morphology of lipid nanoparticles was observed through a transmission electron microscope (TEM). The morphological shape is shown in Fig.

As a result, it was confirmed that the prepared lipid nanoparticles were spherical with diameters ranging from about 100 to 150 nm. This was confirmed to be the result corresponding to the average particle size through the ELS spectrophotometer of Experimental Examples 1, 3, 4 and 8 described above.

Claims (16)

(A) preparing an oil phase by adding Orobol, a solid lipid Cocoa butter or Shea butter to a solvent and dissolving at 40 to 80 ° C;
(B) a step of adding a surfactant-added aqueous phase to the oil phase prepared in the step (a) and homogeneously mixing to form nanoparticles;
And (c) cooling the nanoparticles at room temperature to solidify the solid lipid present in the core of the nanoparticles formed.
delete delete delete The method according to claim 1,
The solvent may be,
But are not limited to, transcutol, ethylacetate, ether, n-hexane, cyclohexane, tetrahydrofuran, methylene, chloride, formamide, dimethylsulfoxide, isopropanol, methanol, ethanol, chloroform, propanol, Wherein the at least one lipophilic compound is at least one selected from the group consisting of glycol, glycerin, 1,2-pentanediol, di-panthenol, dipropylene glycol and acetone.
The method according to claim 1,
The surfactant,
Alkyl Ether Sulfate such as Brij, Poloxamer, Tween, Span, Alcohol Ethoxylate, Low Foaming Surfactant, Wherein the EO-PO block polymer is at least one selected from the group consisting of EO-PO block polymer, polyoxyethylene sorbitan fatty acid ester and PEG.
The method according to claim 1,
The step (b)
At a temperature of 40 to 80 캜.
The method according to claim 1,
The water phase of step (b)
Wherein the water-soluble buffer solution is a water-soluble buffer solution having a pH of 4.0 to 8.5 that does not cause water or skin irritation.
The method according to claim 1,
The nanoparticle formation of step (b)
Characterized in that any one selected from the group consisting of a water-in-oil emulsion, a water-in-oil emulsion, a bass-sonicator, a tip-sonicator, a microfluidizer and an SPG membrane is used. A method for producing nanoparticles.
delete An emulsion comprising the oval ball-containing lipid nanoparticles prepared by the method of claim 1.
delete delete delete delete 12. A cosmetic composition containing the emulsion of claim 11.

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