Composition for transdermal administration of drugs comprising multiple microbubbles

Abstract

The present invention relates to multiple microbubbles and a composition for transdermal drug administration containing the same. Specifically, the invention relates to multiple microbubbles formed by mixing a perfluorocarbon compound and a fatty acid, or a solution containing a perfluorocarbon compound and a lipid, as a carrier for transdermal drug administration, and a composition for transdermal drug administration containing the same.

Description

Technology Field

[0001] The present invention relates to multiple microbubbles and a composition for transdermal drug administration containing the same. Specifically, the invention relates to multiple microbubbles formed by mixing a perfluorocarbon compound and a fatty acid, or a solution containing a perfluorocarbon compound and a lipid, as a carrier for transdermal drug administration, and a composition for transdermal drug administration containing the same. Background Technology

[0003] A drug delivery system (DDS) is a method of transporting micro-sized bubbles containing drugs into the bloodstream within blood vessels to the affected area to enable the drugs to act effectively. In addition to blood vessels, the drug delivery routes of DDS can accurately deliver drugs to the affected area through methods such as delivery via the digestive tract or introduction through the skin.

[0004] Ultrasound-assisted DDSs using microbubbles as a drug delivery medium are attracting attention because the use of microbubbles for drug delivery allows ultrasound to be utilized in technologies that improve drug absorption.

[0005] Since the magnitude, direction, and location of the acoustic radiation pressure applied to bubbles can be controlled by ultrasound, a method utilizing this to capture microbubbles at a desired location is being considered. Furthermore, using ultrasound allows not only for targeting the affected area but also for simultaneously improving drug absorption within biological tissues.

[0006] In this regard, a technique (sonoporation) is also being discussed that destroys bubbles with powerful ultrasound to release drugs inside, forms reversible micro-spaces (cavitation) on the cell surface through micro-jets upon bubble destruction, and efficiently administers drugs into the cell.

[0007] Against this technical background, the inventors of the present application conducted research to find an effective drug delivery system and confirmed that microbubbles formed by mixing a solution containing a perfluorocarbon compound exhibit useful drug delivery properties, thereby completing the present invention.

[0008] Technologies for delivering drugs via microbubbles have been developed in the past. Korean Published Patent No. 2015-0105228 discloses that liquefied inert gas of perfluorobutane can vaporize to form microbubbles. Additionally, Korean Published Patent No. 2019-0110477 discloses ultrasonically responsive microbubbles for drug delivery via a perfluorocarbon-based inert gas.

[0009] However, while these conventional technologies use microbubbles to form reversible microspaces in cells, the present invention is a technology capable of forming reversible microspaces in the dermis.

[0010] In this regard, Korean published patent No. 2016-0140491 discloses that a microspace can be formed by cavitation by ultrasound at a location near the epidermis by including perfluoropentane, perfluorohexane, perfluoromethylcyclohexane, or perfluorooctane as an inert liquefied gas and vaporizing it by ultrasound.

[0011] However, there is an aspect that differs from the present invention, which is a technology for forming microbubbles through perfluoroether while maintaining a structure in which a perfluorocarbon compound is located in the core and a fatty acid or lipid is located in the shell. The problem to be solved

[0013] The object of the present invention is to provide a composition for transdermal administration of a drug comprising microbubbles, characterized in that the microbubbles comprise multiple microbubbles formed by mixing a perfluorocarbon compound and a solution containing fatty acids or lipids, wherein the perfluorocarbon compound comprises methyl perfluoroisobutyl ether. means of solving the problem

[0015] To achieve the above objective, the present invention comprises a first microbubble formed by including a perfluorocarbon compound and a fatty acid; and

[0016] It comprises multiple microbubbles mixed with second microbubbles formed including a perfluorocarbon compound and lipids, and

[0017] The above microbubbles provide a composition for transdermal administration of a drug capable of acoustic droplet vaporization (ADV) by ultrasonic irradiation. Effects of the invention

[0019] The composition for transdermal administration of a drug according to the present invention comprises multiple microbubbles, which can induce an acoustic droplet vaporization phenomenon by ultrasonic energy upon in vivo administration, and can be used as an ultrasound-based drug delivery system. It provides synergistic and booster effects to ultrasound procedures, which are non-invasive transdermal delivery systems, thereby improving penetration efficiency and contributing to the transdermal delivery of active ingredients. Brief explanation of the drawing

[0021] Figure 1 is a schematic diagram showing the manufacturing process of ADV multi-microbubbles. Figure 2(a) shows the result of storing the prepared ADV multi-microbubble solution at room temperature (25℃) for 1 to 7 days, and Figure 2(b) shows the result of ultrasonically irradiating the ADV multi-microbubble solution stored for 7 days. Figure 3 shows a schematic diagram of the US system setup for the aluminum foil experiment. Figure 4 shows photographic images of aluminum foil after treating ADV single microbubbles and multiple microbubbles according to the present invention with ultrasound (20 kHz, 3 W / cm2) for 5 minutes. (A): ultrasound alone, (B): ultrasound combined with water, (C): ultrasound combined with ADV single microbubbles, (D): ultrasound combined with ADV multiple microbubbles. FIG. 5 shows the penetration depths of the control group treated with water and US, the group combining ADV single microbubbles and US, and the group combining ADV multiple microbubbles and US according to the present invention, when the Evans Blue solution was left for 5 minutes. US: Ultrasound. * p <0.05; The data are the mean and SD values. Figure 6 shows the results of analyzing micro-jet velocity and penetration depth according to the type of perfluorocarbon (PFC) and shell. Control: Control group combining ultrasound and water. Fig. 7 shows ultrasound (20 kHz, 3 W / cm²) on pig skin. 2 This shows images and penetration depths of the Evans Blue solution after treatment with (5 min) and MB1 or MB2 solution. Control: Control group combining ultrasound and water, MB: Microbubbles. Figure 8 shows the size of microchannels formed after treating pig skin with ultrasound (20 kHz, 3 W / cm2, 5 min) and MB1 solution (a) and MB2 solution (b), respectively. Specific details for implementing the invention

[0022] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by a skilled expert in the art to which this invention pertains. In general, the nomenclature used herein is well known and commonly used in the art.

[0023] The present invention relates to a composition for transdermal administration of a drug, comprising a first microbubble formed by including a perfluorocarbon compound and a fatty acid; and a multi-microbubble mixed with a second microbubble formed by including a perfluorocarbon compound and a lipid, wherein the microbubbles are capable of acoustic droplet vaporization (ADV) by ultrasonic irradiation.

[0024] Each of the first microbubbles formed including the perfluorocarbon compound and fatty acid; and the second microbubbles formed including the perfluorocarbon compound and lipid is a microbubble capable of acoustic droplet vaporization (ADV) by ultrasonic irradiation.

[0025] The above perfluorocarbon compound may be characterized by having a structure in which it is located in the core, and the fatty acid or lipid is located in the shell.

[0026] "Microbubbles" may refer to gas spheres that generate air or segmented gas. Engineered microbubbles capable of moving freely within blood vessels can apply cavitation effects to drug delivery.

[0027] The interior of the aforementioned microbubbles contains a perfluorocarbon compound. "Perfluoro" stands for "perfluoro / perfluorinated," meaning that all carbon atoms in the molecular chain have been converted into fluorine atoms.

[0028] "Perfluorocarbons (PFCs)" refers to "perfluorocarbons, perfluorocarbons, or perfluorocarbons," which are compounds in which all CH groups in the chain are substituted with CF, and may include PFC precursors in which the perfluoroalkyl moiety is bonded to a non-perfluorinated atom but has the potential to eventually be converted into a PFC.

[0029] According to the present invention, the perfluorocarbon compound preferably comprises a perfluoro ether, and may be, for example, methyl perfluoroisobutyl ether, methyl perfluorobutyl ether, ethyl perfluorobutyl ether, ethyl perfluoroisobutyl ether, polyperfluoromethylisopropyl ether, polyperfluoroisopropyl ether, or a mixture of two or more.

[0030] More preferably, the perfluoroether may be methyl perfluoroisobutyl ether (MPE). There have been no cases of developing a transdermal drug delivery system combined with ultrasound using MPE, which was previously used only as a cosmetic ingredient. MPE is harmless to the human body and is inexpensive, making it suitable for mass production and commercialization.

[0031] The above MPE can be mixed with a solvent, for example, glycol.

[0032] The glycol mentioned above may be, for example, propylene glycol (PG), butylene glycol, or polyethylene glycol, but is not limited thereto.

[0033] In order to produce the first microbubble, the MPE may be included in the solution containing glycol at a concentration of, for example, 1.0 to 3.0% (v / v). Specifically, the MPE may be included in the solution containing glycol at a concentration of 1.5 to 2.5% (v / v) to produce the first microbubble.

[0034] In order to produce the second microbubble, the MPE may be included in the solution containing glycol at a concentration of, for example, 1.0 to 3.0% (v / v). Specifically, the MPE may be included in the solution containing glycol at a concentration of 1.5 to 2.5% (v / v) to produce the second microbubble.

[0035] The above fatty acids may include, for example, stearic acid, myristic acid, palmitic acid, trans fatty acids, cisoleic acid or linoleic acid, or mixtures thereof, but are not limited thereto.

[0036] More preferably, the fatty acid may be, for example, stearic acid.

[0037] The above lipids are, for example, 1,2-dioleoyl-sn-glycero-3-phosphocholine (1,2-dioleoyl-sn-glycero-3-phosphocholine, DOPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, DOPE), 1,2-diheptanoyl-sn-glycero-3-phosphocholine (1,2-diheptanoyl-sn-glycero-3-phosphocholine, DHPC), 1,2-dilauroyl-sn-glycero-3-phosphocholine (1,2-dilauroyl-sn-glycero-3-phosphocholine, DLPC), and 1,2-dipyristoyl-sn-glycero-3-phosphocholine (1,2-dimyristoyl-sn-glycero-3-phosphocholine, DMPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (1,2-dipalmitoyl-sn-glycero-3-phosphocholine, DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (1,2-distearoyl-sn-glycero-3-phosphocholine, DSPC), 1,2-dilinoleoyl-sn-glycero-3-phospho-L-serine (1,2-dilinoleoyl-sn-glycero-3-phosphocholine, DLPC), 1-stearoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (1-stearoyl-2-oleoyl-sn-glycero-3-phospho-L-serine, SOPS), It may be 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), CTAB (cetyltrimethylammonium bromide), or a mixture thereof, but is not limited thereto.

[0038] More preferably, the lipid may include DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) and / or CTAB (cetyltrimethylammonium bromide).

[0039] The above fatty acids or lipids may be mixed with a solvent, for example, glycol.

[0040] The glycol mentioned above may be, for example, propylene glycol (PG), butylene glycol, or polyethylene glycol, but is not limited thereto.

[0041] In some cases, the solution for forming the first microbubble and / or the second microbubble may contain a surfactant. Both anionic and cationic surfactants may be used as the surfactant, and nonionic surfactants may also be used. The above surfactants may be, for example, sodium dodecyl sulfate (SDS), potassium lauryl sulfate, sodium dodecylbenzenesulfonate, sodium laureth sulfate, sodium lauroyl sarcosinate, sodium myreth sulfate, sodium phares sulfate, sodium stearate, ammonium lauryl sulfate, or dioctyl sodium sulfosuccinate, but are not limited thereto.

[0042] The fatty acid may be included in a solution containing glycol to produce the first microbubble at a concentration of, for example, 0.01 to 1.0% (v / v). Specifically, the fatty acid may be included in a solution containing glycol to produce the first microbubble at a concentration of 0.01 to 0.5% (v / v).

[0043] The above lipid may be included in a solution containing glycol for producing the second microbubble at a concentration of, for example, 0.01 to 1.0% (v / v). Specifically, the above lipid may be included in a solution containing glycol for producing the second microbubble at a concentration of 0.01 to 0.1% (v / v).

[0044] Specifically, in an embodiment according to the present invention, the fatty acid is stearic acid, and the lipid may include DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) and / or CTAB (cetyltrimethylammonium bromide).

[0045] Each of the above fatty acids and lipids is mixed in butylene glycol and may be included in the solution at concentrations of 0.01 to 0.5% (v / v) and 0.01 to 0.1% (v / v), respectively.

[0046] The first microbubble and the second microbubble are mixed to form multiple microbubbles. It includes a first microbubble composed of a stearic acid shell and a second microbubble composed of a DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) shell. The first microbubble and the second microbubble both contain MPE (Methyl perfluoroisobutyl ether) inside.

[0047] The first microbubble and the second microbubble may be mixed in a concentration ratio of 7.5:2.5 to 9.5:0.5. Specifically, the first microbubble and the second microbubble may be mixed in a concentration ratio of 8:2 to 9:1, and more specifically, the first microbubble and the second microbubble may be mixed in a concentration ratio of 9:1.

[0048] The microbubbles according to the present invention can undergo acoustic droplet vaporization (ADV) by ultrasonic irradiation. Specifically, a core portion containing perfluorocarbon can induce an acoustic droplet vaporization phenomenon by ultrasonic waves.

[0049] Ultrasound refers to sound waves with frequencies higher than the range of sounds audible to humans.

[0050] The above ultrasound may have a frequency of, for example, 20 kHz to 3 MHz. The frequency of ultrasound generally used for therapeutic purposes is 1 to 3 MHz, and the frequency used in aesthetics is mostly 20 to 50 kHz.

[0051] As the frequency increases, the resonance size may decrease, and taking this into account, the minimum particle size of the microbubble may be 4 or larger. If the microbubble has a suitable size that responds to the ultrasonic frequency, vaporization can occur more effectively.

[0052] Taking this into consideration, the microbubbles according to the present invention are 4 μm or larger, 15 m It can have a particle size of m or less.

[0053] "Acoustic droplet vaporization (ADV)" refers to the process of vaporizing a superheated droplet emulsion into bubbles using ultrasound. Acoustic cavitation can be considered as one of the theories explaining the physical mechanism of the acoustic droplet vaporization.

[0054] Cavitation can generally be classified into stable cavitation and inertial cavitation. Stable cavitation occurs when bubbles repeatedly contract and expand due to low acoustic pressure amplitude. When this phenomenon occurs at an interface, microstreaming takes place, inducing shear stress, which can enhance drug penetration efficiency through biological barriers such as cell membranes.

[0055] In contrast, inertial cavitation refers to the phenomenon where bubbles rapidly expand and then collapse upon exposure to high-amplitude ultrasound. This bubble collapse occurs asymmetrically near the interface, and the resulting microjet temporarily reduces the barrier function of the interface, thereby improving drug penetration efficiency.

[0056] As such, cavitation is a major mechanism of ultrasound-based drug delivery technology, and microbubbles can serve as key drug carriers to implement this.

[0057] Based on this, the present invention is a delivery vehicle for transdermal administration of a drug containing microbubbles, wherein the drug may be mixed in a solution containing a first microbubble and / or a second microbubble.

[0058] Non-invasive transdermal drug delivery is possible through the present invention. By applying ultrasonic energy to promote the transdermal absorption of a drug applied to the skin, rapid and easy drug administration can be provided. Since the necessary amount of drug to maximize efficacy and effect can be administered non-invasively to the affected area, not only can side effects be reduced, but the accelerated absorption during administration can also rapidly enhance the therapeutic effect of the drug.

[0059] The above drug may include, for example, a functional cosmetic ingredient or a pharmaceutical ingredient as an active ingredient for transdermal administration.

[0060] The above composition may additionally include 0.01 to 5 weight percent of a functional cosmetic ingredient or a pharmaceutical ingredient based on the total amount of the multi-microbubble composition.

[0061] At this time, the functional cosmetic ingredient may include, for example, an active ingredient for whitening, wrinkle improvement, UV protection, cell regeneration, moisturizing, elasticity, exfoliation, sebum removal, or the prevention or treatment of atopic dermatitis or acne.

[0062] The above medicinal ingredients may include, for example, ingredients for relieving inflammation or pain, providing cooling and warming effects, or for wound healing.

[0064] The present invention will be described in more detail below through examples. These examples are intended solely to illustrate the present invention, and it will be obvious to those skilled in the art that the scope of the present invention is not to be interpreted as being limited by these examples.

[0066] example

[0068] [Preparation Example] Preparation of ADV (Acoustic Droplet Vaporization) Multi-Microbubble Solution

[0069] The ADV multi-microbubble solution comprises a first microbubble composed of a stearic acid membrane (shell) and a second microbubble composed of a DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) membrane (shell). Both the first and second microbubbles contain MPE (Methyl perfluoroisobutyl ether) internally. [Fig. 1]

[0070] A first microbubble solution was prepared by adding and mixing 50 mL of distilled water, 10 mL of butylene glycol, 0.5% (w / v) of stearic acid, and 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). A second microbubble solution was prepared by adding 0.1% (w / v) of DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) to 50 mL of distilled water, heating and mixing at 80°C, and then adding and stirring 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). Next, the first microbubble solution and the second microbubble solution were mixed in a 9:1 ratio to prepare an ADV multi-microbubble solution.

[0072] [Example 1] Storage Stability

[0074] To determine storage conditions, a stability test was conducted at room temperature (25℃). 3.5ml of a multi-microbubble solution stored at room temperature was dispensed into a silicone tube with a volume of 4ml, and then placed at a temperature similar to human body temperature (36℃). 20 kHz, 3 W / cm² 2 The vaporization of microbubbles was confirmed by treating with ultrasound under certain conditions.

[0075] As shown in Figure 2, it was confirmed that the multiple microbubbles in the ADV solution were still vaporized even after being stored at room temperature for 7 days, which indicates that the storage stability of ADV multiple microbubbles is superior compared to SonoVue, which has a stability of 6 hours.

[0077] [Example 2] Aluminum Foil Penetration Analysis

[0079] (1) Preparation of multi-microbubble solution using ADV (Acoustic Droplet Vaporization)

[0080] The ADV multi-microbubble solution comprises a first microbubble composed of a stearic acid shell and a second microbubble composed of a DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) shell. Both the first microbubble and the second microbubble contain MPE (Methyl perfluoroisobutyl ether) inside.

[0081] A first microbubble solution was prepared by adding and mixing 50 mL of distilled water, 10 mL of butylene glycol, 0.5% (w / v) of stearic acid, and 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). A second microbubble solution was prepared by adding 0.1% (w / v) of DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) to 50 mL of distilled water, heating and mixing at 80°C, and then adding and stirring 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). Next, the first microbubble solution and the second microbubble solution were mixed in a 9:1 ratio to prepare an ADV multi-microbubble solution.

[0082] The ADV single-microbubble used in this experiment refers to a first microbubble composed of a stearic acid shell.

[0084] (2) Aluminum foil

[0085] The aluminum foil used is 100 m m thickness, 5 x 5 cm 2 I cut it into the appropriate size and used it.

[0087] (3) Aluminum Foil Penetration Assay

[0088] Aluminum foil penetration analysis was performed using a modified method of Wolloch L., Kost J., The importance of microjet vs shock wave formation in sonophoresis, J. Control. Release, 148(2) 204-211 (2010). Thickness 100 m m, 5 x 5 cm 2 Add ADV single and multi-microbubbles to aluminum foil prepared to size, then 20 kHz, 3 W / cm 2 The condition was treated with ultrasound for 5 minutes. 1 x 1 cm 2 A square plate with holes of a certain size was used, and to prevent leakage of the ADV microbubble solution, an ultrasonic gel was used to form a square border, after which the solution was added and ultrasonic treatment was performed. After ultrasonic treatment, the ultrasonic gel was washed and removed, and the diameter of the holes formed in the aluminum foil was measured. (Fig. 3)

[0090] (4) Result

[0092] As a result of measuring the diameter of the holes formed in the aluminum foil using an aluminum foil penetration assay, pores measuring 3.23 ± 0.15 mm and 9.16 ± 0.25 mm were identified in the groups treated simultaneously with US and ADV single and multiple microbubbles, respectively. In addition, no traces of penetration or pores were observed in the US and the groups treated with US and water. (Fig. 4)

[0093] The group treated with US and ADV multi-microbubbles showed a pore size more than 2.83 times larger compared to the group treated with US and ADV single-microbubbles. This suggests that ADV multi-microbubbles can contribute to microchannel formation more than ADV single-microbubbles and can improve the penetration efficiency of active ingredients.

[0094] [Table 1] Pore size formed in aluminum foil

[0095]

[0097] [Example 3] Agarose phantom penetration assay

[0099] (1) Preparation of multi-microbubble solution using ADV (Acoustic Droplet Vaporization)

[0100] The ADV multi-microbubble solution comprises a first microbubble composed of a stearic acid shell and a second microbubble composed of a DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) shell. Both the first microbubble and the second microbubble contain MPE (Methyl perfluoroisobutyl ether) inside.

[0101] A first microbubble solution was prepared by adding and mixing 50 mL of distilled water, 10 mL of butylene glycol, 0.5% (w / v) of stearic acid, and 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). A second microbubble solution was prepared by adding 0.1% (w / v) of DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) to 50 mL of distilled water, heating and mixing at 80°C, and then adding and stirring 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). Next, the first microbubble solution and the second microbubble solution were mixed in a 9:1 ratio to prepare an ADV multi-microbubble solution.

[0102] The ADV single-microbubble used in this experiment refers to a first microbubble composed of a stearic acid shell.

[0104] (2) Manufacture of Agarose Phantom

[0106] The agarose phantom used was prepared by adding agarose, distilled water, 70% ethanol, and NaCl (sodium chloride) to a beaker, respectively, and then heating and stirring at 120°C. After stirring, the dissolved liquid was cooled to a gel state to produce an agarose phantom (diameter 1 cm, height 2 cm). (Table 2)

[0107] [Table 2] Composition of Agarose Phantom Solution

[0108]

[0110] (3) Agarose phantom penetration assay

[0111] The agarose phantom penetration assay was conducted by modifying the method of Liao AH, Ma WC, Wang CH, and Yeh MK, Penetration depth, concentration and efficiency of transdermal α-arbutin delivery after ultrasound treatment with albumin-shelled microbubbles in mice, Drug Deliv., 23(7) 2173-2182 (2016). ADV single microbubbles and 600 ADV multi-microbubble solution according to the present invention were applied to an agarose phantom. m After applying L to each, ultrasound (20 kHz, 3 W / cm²) 2 ) was treated for 5 minutes. After sonication, Evans Blue (0.1 mg / mL) 200 m L was added to the sonication area and left to absorb for 5 minutes. Then, the penetration depth of the Evans Blue solution was measured using a fluorescence microscope (DMIL LED Fluo, Leica Co. Ltd., Germany).

[0113] (4) Agarose phantom penetration measurement results

[0114] As a result of measuring the penetration depth of Evans Blue solution using the agarose phantom penetration assay, the control group treated with ultrasound (US) and water was 12.68 ± 1.08 m The group treated with single microbubbles in m, US and ADV was 30.73 ± 2.56 m The group treated with m, US and ADV multi-microbubbles was 78.12 ± 1.75 m The penetration depth of m was shown (Table 3, Fig. 5).

[0115] The group treated with multiple microbubbles of US and ADV simultaneously showed penetration efficiencies of 6.16 and 2.54 times, respectively, compared to the control group treated with US and water and the group treated with single microbubbles of US and ADV.

[0116] ADV multi-microbubbles have superior penetration efficiency compared to ADV single-microbubbles, which can improve penetration efficiency by providing a booster effect to ultrasound procedures used for transdermal delivery.

[0118] [Example 4] Microjet velocity and penetration depth according to shell type

[0119] (1) Preparation of a Surfactant-Microbubble composed of a surfactant shell

[0120] Surfactant shell-microbubbles composed of a surfactant membrane were prepared by adding 50 mL of distilled water, 10 mL of butylene glycol, and 0.1% (v / v) of CTAB (cetyltrimethylammonium bromide), followed by MPE (methyl perfluoroisobutyl ether) and PFB (perfluorobutane, C4F 10 ), PFH(PerfluoroHexane, C6F 14 It was prepared by adding and stirring 2.5% (v / v).

[0122] (2) Preparation of Lipid-Microbubble composed of a lipid shell

[0123] Lipid shell microbubbles composed of lipid membranes were prepared by adding 50 mL of distilled water, 10 mL of butylene glycol, and 0.1% (w / v) of DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine), heating and mixing at 80°C, and then preparing PFB (PerfluoroButane, C4F 10 ), PFH(PerfluoroHexane, C6F 14 It was prepared by adding and stirring 2.5% (v / v).

[0125] (3) Preparation of acrylamide gel

[0126] A 40% acrylamide gel solution was diluted with distilled water, and then 10% ammonium persulfate and TEMED (Tetramethylethylenediamine) were added as polymerization initiators to prepare a 20% acrylamide gel.

[0128] (4) Analysis of micro-jet velocity and penetration depth

[0129] 600 μL of each prepared microbubble was added to a 20% acrylamide gel, followed by ultrasound (20 kHz, 3 W / cm²). 2 ) was treated for 5 minutes. The micro-jet velocity of the microbubbles was confirmed by analyzing images captured at a high frame rate using a high-speed camera. After sonication, 150 of 0.25% bromophenol blue solution m L was added to the ultrasonic treatment area and left to be absorbed for 5 minutes to analyze the penetration depth.

[0131] (5) Analysis of micro-jet velocity and penetration depth according to perfluorocarbon and shell type

[0132] The correlation between the micro-jet velocity and penetration depth of microbubbles according to the type of perfluorocarbon and shell was analyzed. As a result of analyzing the micro-jet velocity for surfactants and lipid membranes, microbubbles composed of lipid membranes exhibited a micro-jet velocity approximately 1.3 times higher than microbubbles composed of surfactant membranes.

[0133] The microjet velocity of the Lipid shell-MPE group was the highest at 190.14 m / s, and the penetration depth was measured at 374.75 μm (Fig. 6).

[0134] As a result of analyzing the penetration depth according to the type of perfluorocarbon, it was confirmed that the Lipid shell-MPE group had a penetration depth 3 times and 1.05 times greater than that of the Lipid shell-PFB group and the Lipid shell-PFH group, respectively.

[0135] A lipid shell thicker than a surfactant shell can provide a faster microjet velocity. Additionally, faster microjet velocities increase penetration depth, which can contribute to improved penetration efficiency and transdermal delivery.

[0137] [Example 5] Microchannel formation and penetration effects according to shell type

[0139] (1) MB1 manufacturing

[0140] The ADV multi-microbubble solution composed of a surfactant membrane comprises a first microbubble composed of a stearic acid membrane (shell) and a second microbubble composed of a CTAB (cetyltrimethylammonium bromide) membrane (shell). Both the first microbubble and the second microbubble contain MPE (Methyl perfluoroisobutyl ether) inside.

[0141] A first microbubble solution was prepared by adding and mixing 50 mL of distilled water, 10 mL of butylene glycol, 0.5% (w / v) of stearic acid, and 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). A second microbubble solution was prepared by adding 0.1% (w / v) of CTAB (cetyltrimethylammonium bromide) to 50 mL of distilled water, heating and mixing at 80°C, and then adding and stirring 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). Next, the first microbubble solution and the second microbubble solution were mixed in a 9:1 ratio to prepare an ADV multi-microbubble solution.

[0143] (2) MB2 manufacturing

[0144] The ADV multi-microbubble solution composed of a lipid membrane comprises a first microbubble composed of a stearic acid membrane (shell) and a second microbubble composed of a DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) membrane (shell). Both the first microbubble and the second microbubble contain MPE (Methyl perfluoroisobutyl ether) inside.

[0145] A first microbubble solution was prepared by adding and mixing 50 mL of distilled water, 10 mL of butylene glycol, 0.5% (w / v) of stearic acid, and 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). A second microbubble solution was prepared by adding 0.1% (w / v) of DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) to 50 mL of distilled water, heating and mixing at 80°C, and then adding and stirring 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). Next, the first microbubble solution and the second microbubble solution were mixed in a 9:1 ratio to prepare an ADV multi-microbubble solution.

[0147] (3) Prepare Porcine skin

[0148] Pig skin purchased from the slaughterhouse was used within one week of freezing (-20℃). After slowly thawing at room temperature, 2.5 x 2.5 cm 2 , cut to a thickness of 2mm and used.

[0150] (4) Porcine skin penetration assay

[0151] Porcine skin penetration analysis was performed on 1,350 MB1 and MB2 solutions on fixed porcine skin. mAfter applying L to each, ultrasound (20 kHz, 3 W / cm²) 2 ) was treated for 5 minutes. After sonication, 150 Evans Blue solution (0.25 mg / mL) m L was added to the sonication area and left to absorb for 5 minutes. Then, after washing and removing the treated Evans Blue solution, the penetration depth of the Evans Blue solution was measured using a fluorescence microscope (DMIL LED Fluo, Leica Co. Ltd., Germany).

[0153] (5) Porcine skin penetration measurement results

[0154] As a result of measuring the penetration depth of Evans Blue solution into porcine skin, the control group treated with ultrasound (US) and water was 20.59 ± 0.65 m The group treated with m, US, and MB1 was 166.37 ± 4.29 m The group treated with m, US, and MB2 was 272.11 ± 3.35 m The penetration depth of m was shown (Fig. 7).

[0155] The group treated with MB2, which has a shell composed of US and DOPE, showed superior penetration efficiency of 13.21 and 1.64 times, respectively, compared to the control group treated with US and water and the group treated with MB1, which has a shell composed of US and CTAB.

[0156] The longer the length of the chains constituting the shell, the more it improves penetration efficiency and can contribute to the transdermal delivery of active ingredients.

[0158] [Example 6] Microchannel formation according to shell type

[0159] (1) MB1 manufacturing

[0160] The ADV multi-microbubble solution composed of a surfactant membrane comprises a first microbubble composed of a stearic acid membrane (shell) and a second microbubble composed of a CTAB (cetyltrimethylammonium bromide) membrane (shell). Both the first microbubble and the second microbubble contain MPE (Methyl perfluoroisobutyl ether) inside.

[0161] A first microbubble solution was prepared by adding and mixing 50 mL of distilled water, 10 mL of butylene glycol, 0.5% (w / v) of stearic acid, and 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). A second microbubble solution was prepared by adding 0.1% (w / v) of CTAB (cetyltrimethylammonium bromide) to 50 mL of distilled water, heating and mixing at 80°C, and then adding and stirring 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). Next, the first microbubble solution and the second microbubble solution were mixed in a 9:1 ratio to prepare an ADV multi-microbubble solution.

[0163] (2) MB2 manufacturing

[0164] The ADV multi-microbubble solution composed of a lipid membrane comprises a first microbubble composed of a stearic acid membrane (shell) and a second microbubble composed of a DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) membrane (shell). Both the first microbubble and the second microbubble contain MPE (Methyl perfluoroisobutyl ether) inside.

[0165] A first microbubble solution was prepared by adding and mixing 50 mL of distilled water, 10 mL of butylene glycol, 0.5% (w / v) of stearic acid, and 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). A second microbubble solution was prepared by adding 0.1% (w / v) of DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine) to 50 mL of distilled water, heating and mixing at 80°C, and then adding and stirring 2.5% (v / v) of MPE (Methyl perfluoroisobutyl ether). Next, the first microbubble solution and the second microbubble solution were mixed in a 9:1 ratio to prepare an ADV multi-microbubble solution.

[0167] (3) Prepare Porcine skin

[0168] Pig skin purchased from the slaughterhouse was used within one week of freezing (-20℃). After slowly thawing at room temperature, 2.5 x 2.5 cm 2 , cut to a thickness of 2mm and used.

[0170] (4) SEM pretreatment of pig skin

[0171] The ADV ultrasonically treated pig skin was fixed with 2.5% glutaraldehyde and then washed with PBS (Phosphate Buffered Saline). After dehydration while increasing the ethanol concentration from 25% to 100%, the skin was dried using a critical point dryer (Bal-Tec, Germany) and then photographed with SEM (Hitachi Ltd., Tokyo, Japan).

[0173] (5) SEM analysis of pig skin

[0174] As a result of measuring the size of microchannels formed in porcine skin, the group treated with US and MB1 had a diameter of 12 μm, whereas the group treated with US and MB2 formed microchannels with a maximum diameter of 33 μm. (Fig. 8)

[0175] The group treated with MB2, which consists of US and DOPE shells, formed microchannels that were more than 2.75 times larger than the group treated with MB1, which consists of US and CTAB shells.

[0176] The longer the length of the chains constituting the shell of multiple microbubbles, the larger the size of the formed microchannels, suggesting that this can contribute to the transdermal delivery efficiency of the active ingredient.

[0178] Foregoing, specific parts of the content of the present invention have been described in detail. It will be apparent to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the present invention. Accordingly, the actual scope of the present invention is defined by the appended claims and their equivalents.

Claims

Claim 1 A composition for transdermal administration of a drug, comprising a first microbubble formed by including i) methyl perfluoroisobutyl ether and ii) stearic acid; and a multi-microbubble mixed with a second microbubble formed by including i) methyl perfluoroisobutyl ether and ii) DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine), wherein the microbubbles are capable of acoustic droplet vaporization (ADV) by ultrasonic irradiation. Claim 2 delete Claim 3 delete Claim 4 delete Claim 5 A composition according to claim 1, characterized in that the ultrasound has a frequency of 20 kHz to 3 MHz.

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