Ionic liquid emulsion as well as preparation method and application thereof in medicine permeation and delivery

Abstract

The invention discloses an ionic liquid emulsion, a preparation method thereof and application of the ionic liquid emulsion in medicine permeation and delivery, and belongs to the technical field of transdermal penetration. The ionic liquid emulsion is composed of a water phase and an oil phase in a volume ratio of (9-7): (1-3), the water phase is an ionic liquid containing organic acid and a sophora flavescens total alkaloid extract, the oil phase is an emulsion containing a transdermal penetration enhancer, a solvent and hydroxypropyl methyl cellulose, and the preparation method comprises the following steps: (1) dissolving the hydroxypropyl methyl cellulose in the solvent, stirring and mixing, adding the transdermal penetration enhancer, and stirring to obtain a mixture; shearing and homogenizing to obtain an emulsion; (2) dissolving organic acid and the sophora flavescens total alkaloid extract in a solvent for reaction, and removing the solvent after the reaction is finished to prepare an ionic liquid; and (3) dissolving the ionic liquid in the emulsion to obtain the ionic liquid. According to the invention, the ionic liquid emulsion and ultrasound are combined for promoting permeation, so that the permeation effect of the medicine on deep skin can be obviously enhanced.

Description

Technical Field

[0001] The invention belongs to the field of transdermal technology, and in particular relates to an ionic liquid emulsion and a preparation method thereof, and an application thereof in drug penetration and delivery. Background Art

[0002] Transdermal drug delivery refers to the delivery of drugs to the site of action through skin absorption to achieve systemic or local therapeutic effects, and is used to prevent or treat diseases. Common dosage forms include patches, paints, ointments, plasters, powders, and cataplasms. Compared with other methods, transdermal drug delivery avoids the effects of digestion and metabolic tissues on drug efficacy, acts directly on target tissues, and has the characteristics of rapid onset and stable drug efficacy of local administration. These advantages make the market potential of transdermal drug delivery technology huge. However, the research and development of transdermal preparations still faces urgent problems to be solved. For example, the penetration depth of transdermal preparations is limited, and they are often accompanied by skin allergies or low blood drug concentrations. Therefore, finding excellent permeation-promoting methods to allow drugs to pass through the skin more effectively is a more mainstream research direction.

[0003] At present, the following strategies can be used to promote transdermal penetration: 1. Physical penetration enhancers. For example, iontophoresis is the process of ionic drugs passing through the skin under the action of an electric field. This method is mostly used in the study of drug delivery systems for biomacromolecules such as proteins and proteins. Electroporation is a method of using an instantaneous high-voltage pulse electric field to form temporary, reversible hydrophilic channels in lipid bilayers such as cell membranes to increase permeability. Ultrasonic introduction, also known as ultrasonic introduction, is a method of using ultrasonic energy to promote transdermal penetration of drugs. Microneedle technology forms a micron-scale hole array on the stratum corneum of the skin through the puncture of microneedles, thereby achieving drug introduction. Needle-free jet drug delivery, laser-promoted transdermal drug delivery, thermal energy-promoted transdermal drug delivery, etc. However, physical penetration enhancement methods also have certain advantages and disadvantages. Taking patches as an example, it is found that the active ingredients of the drugs in the patches are not only released when applied, but also continue to be released after the patches are removed, which is a great factor affecting skin irritation. Therefore, in order to reduce irritation, it is best to find a way to remove the residual drugs after the patches are removed. 2. Chemical penetration enhancers (CPEs). Chemical penetration enhancers include water, azone, sulfoxide and its analogs, surfactants, ethanol, fatty alcohols, pyrrolidines, natural transdermal penetration enhancers, etc. The traditional mechanism of action of CPEs is to change the skin properties, disrupt the arrangement of lipids, form gaps in tight connections, and thus increase the water content of the stratum corneum. Its penetration enhancement process is non-specific, and high concentrations or long-term use can cause adverse reactions such as skin irritation and allergic reactions. CPEs have defects such as low biocompatibility and high irritation. 3. Biological penetration enhancers mainly include the following categories: penetration peptides, metabolic regulators, amino acid derivatives, and ceramides and their analogs. However, before widespread application, at least the following problems need to be solved: establishment of in vitro and in vivo correlation, drugability, establishment of dose-effect relationship, safety issues, etc. 4. Pharmaceutical methods, such as microemulsions, liposomes, nanoparticles, etc. This method has certain advantages and disadvantages. The advantages are to improve drug permeability, adapt to specific treatment needs, and enhance drug stability. The disadvantages are that it may cause skin irritation, cost issues, safety issues, etc. Summary of the invention

[0004] Purpose of the invention: In order to solve the problems existing in the prior art, the first purpose of the present invention is to provide an ionic liquid emulsion with excellent penetration-promoting effect and antibacterial performance, the second purpose of the present invention is to provide a method for preparing the above-mentioned ionic liquid emulsion, and the third purpose of the present invention is to provide the application of the above-mentioned ionic liquid emulsion in drug penetration and delivery.

[0005] Technical solution: The ionic liquid emulsion of the present invention is composed of an aqueous phase and an oil phase in a volume ratio of 9-7:1-3, wherein the aqueous phase is an ionic liquid containing an organic acid and a total alkaloid extract of Sophora flavescens, and the oil phase is an emulsion including a transdermal penetration enhancer, a solvent and hydroxypropyl methylcellulose.

[0006] Furthermore, the total alkaloid extract of Sophora flavescens includes matrine, oxymatrine, sophoridine, sophorocarpine or oxysophorocarpine.

[0007] Furthermore, the transdermal penetration enhancer is one or more of lavender essential oil, rosemary essential oil, peppermint essential oil, eucalyptus essential oil, tea tree essential oil, geranium essential oil, frankincense essential oil, myrrh essential oil, carrot seed essential oil, sweet orange essential oil, lemon essential oil, grapefruit essential oil, ylang ylang essential oil, patchouli essential oil, sandalwood essential oil, basil essential oil, fennel essential oil, cinnamon essential oil, clove essential oil, and ginger essential oil.

[0008] Preferably, the transdermal penetration enhancer is eucalyptus essential oil, that is, eucalyptol, which is extracted from the leaves of eucalyptus and is the main active ingredient of lavender oil, tea tree essential oil, atractylodes oil, myrtle essential oil, etc. It has the effects of dispelling wind and relieving heat, inhibiting bacteria and inflammation, antiseptic and antipruritic, antioxidant and insecticidal and antipruritic. At the same time, eucalyptus oil also has a good transdermal penetration enhancing effect and is an additive for many skin medication preparations and cosmetics.

[0009] Furthermore, the molar ratio of the organic acid to the total alkaloid extract of Sophora flavescens is 2-1:1-2, preferably 2:1; and the organic acid is citric acid.

[0010] Furthermore, the dosage ratio of the transdermal penetration enhancer, the solvent and hydroxypropyl methylcellulose is 5-15ml:45-35ml:1.35-1.05g; and the solvent is water.

[0011] The present invention provides a method for preparing the above-mentioned ionic liquid emulsion, comprising the following steps:

[0012] (1) dissolving hydroxypropyl methylcellulose in a solvent, stirring and mixing, adding a transdermal penetration enhancer, shearing, and homogenizing to obtain an emulsion;

[0013] (2) dissolving an organic acid and a total alkaloid extract of Sophora flavescens in a solvent for reaction, and removing the solvent after the reaction to obtain an ionic liquid;

[0014] (3) Dissolving the ionic liquid in the emulsion to obtain the ionic liquid emulsion.

[0015] Furthermore, in step (1), the stirring and mixing conditions are: stirring at 35-40°C for 10-15 minutes; the shearing conditions are: shearing at 8000-10000 rpm for 5-10 minutes; and the homogenizing conditions are: homogenizing at a homogenizing pressure of 460-500 bar for 5-20 minutes, and circulating 5-20 times.

[0016] The present invention provides application of the ionic liquid emulsion in drug penetration and delivery.

[0017] Furthermore, the ionic liquid emulsion is subjected to drug penetration and delivery under ultrasound, and the ultrasound conditions are: ultrasound frequency is 1-2 MHz, ultrasound power is 0.8-1 W, and ultrasound time is 10-30 min.

[0018] Furthermore, the penetration depth is 0-1 mm.

[0019] Principle of the invention: The present invention combines pharmaceutical penetration enhancers (organic acid matrine-based ionic liquids), chemical penetration enhancers (transdermal penetration enhancers), and physical penetration enhancement technology (ultrasound) to enhance the penetration strength and depth of drugs on the skin.

[0020] Sophora flavescens-based ionic liquids have a certain transdermal penetration enhancing effect and good biocompatibility. Sophora flavescens total alkaloid extracts include, for example, matrine, oxymatrine, sophoridine, sophorocarpine, oxysophorocarpine, etc., all of which are alkaloids in Sophora flavescens. However, since ionic liquids are in a liquid state and are inconvenient to administer, and the depth of transdermal penetration of existing ionic liquids is a major problem, the present invention selects a transdermal penetration enhancer as an oil phase to prepare an ionic liquid emulsion.

[0021] Ultrasound increases skin permeability by stimulating lipids in the stratum corneum through the ultrasonic cavitation effect. In addition, the thermal effect can also play an additional role in increasing the permeability by increasing the kinetic energy and diffusion rate of the drug, dilating hair follicles and sweat glands, and enhancing blood circulation in the treatment area.

[0022] Beneficial effects: Compared with the prior art, the present invention has the following significant effects: the present invention utilizes organic acid and matrine-based ionic liquid emulsions to significantly enhance the skin penetration effect under ultrasonic conditions. Compared with other transdermal technologies, the ionic liquid emulsion of the present invention also greatly enhances the drug penetration effect on the deep skin, thereby broadening the medical use of ionic liquid emulsions, providing new ideas for subsequent transdermal penetration enhancement therapy, and having important clinical significance. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The graphs are the changes in droplet size of the emulsion in Example 1 before (A) and after (B) high-pressure homogenization;

[0024] Figure 2 This is a graph showing the change in particle size of the emulsion in Example 1 after high pressure homogenization at different cycles;

[0025] Figure 3 This is the laser confocal microscopy image of the emulsion in Example 1;

[0026] Figure 4 The microstructure of the emulsion in Example 1 after storage for 0 days (freshly prepared), 1 day, 3 days, 5 days, and 10 days;

[0027] Figure 5This is a statistical diagram of the particle size of the ionic liquid emulsion changing with time at different water-to-oil ratios in Example 1;

[0028] Figure 6 This is a diagram showing the transdermal penetration enhancement effect of the Chinese medical ultrasonic coupling agent, the smear emulsion, the smear ionic liquid emulsion, the ultrasonic emulsion, and the ultrasonic ionic liquid emulsion on FITC-Dextran with a molecular weight of 5000 in Example 1;

[0029] Figure 7 This is a graph of FITC fluorescence intensity in the skin after different treatments in Example 1, Scale bar: 0.2 mm;

[0030] Figure 8 This is a fluorescence co-localization analysis diagram after different treatments in Example 1;

[0031] Fig. 9 This is an analysis chart of the change in fluorescence intensity from top to bottom of the skin after different treatments in Example 1. DETAILED DESCRIPTION

[0032] The present invention is further described in detail below in conjunction with the embodiments and drawings.

[0033] Example 1: The ionic liquid emulsion provided in this example consists of an aqueous phase and an oil phase, wherein the aqueous phase is an ionic liquid containing citric acid and matrine, and the oil phase is an emulsion containing eucalyptol, a solvent and hydroxypropyl methylcellulose. The specific experimental process is as follows:

[0034] 1.1 Experimental Materials and Instruments

[0035] 1.1.1 Experimental Materials

[0036] Citric acid, matrine, FITC-Dextran (MW.5K), eucalyptol, hydroxypropyl methylcellulose (HPMC), fluorescein sodium, Nile red, medical ultrasound coupling agent.

[0037] 1.1.2 Experimental instruments

[0038] One-ten-thousandth balance, one-hundred-thousandth balance, heating stirrer, shearing instrument, high-pressure homogenizer, inverted microscope, laser confocal microscope, high-speed centrifuge, 10cm culture dish, plate-coated beads, and bacterial constant temperature incubator.

[0039] 1.1.3 Experimental animals

[0040] Male BALB / c-nu nude mice aged 6-8 weeks were selected.

[0041] 1.2 Experimental methods

[0042] 1.2.1 Preparation of emulsion

[0043] 1.2 g of HPMC was weighed into a 100 ml beaker, 40 ml of purified water was added thereto, and the mixture was heated and stirred at 40 ° C for 10 min. After 10 min, 10 ml of eucalyptol was added thereto, and the mixture was sheared at 8000 rpm for 5 min with a shearing instrument, and then homogenized at 460 bar for 5 min (i.e., homogenization cycle 5 times) by a high-pressure homogenizer to obtain an emulsion.

[0044] 1.2.2 Preparation of ionic liquid emulsion

[0045] The citrate matrine ionic liquid is mixed and dissolved in the above emulsion so that the ionic liquid accounts for 5% of the total system volume. That is, when preparing 10 ml of ionic liquid emulsion, 500 μl of ionic liquid is taken and mixed with 9.5 ml of emulsion to obtain the ionic liquid emulsion.

[0046] 1.2.3 Preparation of emulsions under different homogenization conditions

[0047] 1.2g of HPMC was weighed into a 100ml beaker, 40ml of purified water was added, and the mixture was heated and stirred at 40°C for 10min. After 10min, 10ml of eucalyptol was added, and the mixture was sheared at 8000rpm for 5min. Then, the mixture was homogenized at 460bar for 5min (i.e., 5 homogenization cycles), 10 times, and 20 times by a high-pressure homogenizer to obtain an emulsion. The emulsions with different cycles were photographed and their fluidity was considered.

[0048] 1.2.4 Emulsion laser confocal microscopy images

[0049] To observe the microstructure of the emulsion, laser confocal scanning was performed on the emulsion prepared under the optimal conditions, i.e., water-oil ratio 8:2, heating at 40°C, and high pressure homogenization cycle 5 times. During the preparation process, the water phase was labeled with sodium fluorescein and the oil phase was labeled with Nile red.

[0050] 1.2.5 Emulsion stability assessment

[0051] Emulsions with different water-oil ratios were prepared, with water-oil ratios of 9:1, 8:2, and 7:3. The microstructures were observed on day 0 (freshly prepared), 1 day, 3 days, 5 days, and 10 days later. Stability was evaluated by observing whether stratification occurred and by particle size and uniformity. The particle size was quantitatively calculated using Image J.

[0052] 1.2.6 In vivo transdermal effect of ionic liquid emulsion under ultrasound

[0053] BALB / c-nu nude mice aged 6-8 weeks were selected to compare the transdermal penetration enhancement effects of medical ultrasonic coupling agent, smear emulsion, smear ionic liquid emulsion, ultrasonic emulsion, and 5000 molecular weight FITC-Dextran in ultrasonic ionic liquid emulsion. First, the nude mice were anesthetized, and 1 ml of the above reagents were applied to their backs. Ultrasound was used when ultrasound was needed. The ultrasonic power and time were 1 MHz, 1 W, and 30 min. The residual preparations on the surface were wiped off with PBS. The mice were killed by cervical dislocation. The mouse skin was taken and washed repeatedly in PBS and stored in paraformaldehyde in the dark. Paraffin sections were prepared by paraffin embedding, and the cell nuclei were stained with DAPI. The sections were examined under a fluorescence microscope. The n of each group was 3. Then, HE-stained sections were made to detect whether the above reagents and ultrasonic treatment would cause certain damage to the mouse skin surface.

[0054] 1.2.7 In vitro antibacterial activity of ionic liquid emulsion under ultrasound

[0055] The in vitro antibacterial ability of ultrasound, emulsion, ionic liquid emulsion, ultrasound emulsion, and ultrasound ionic liquid emulsion was evaluated by the plate coating method. The concentration of the Malassezia bacterial suspension was 1×10^6 CFU, and 100μl of bacterial suspension was added to each culture dish. The antibacterial effect was verified by the above operations. The emulsion and ionic liquid emulsion bacteria were 100μl, and the ultrasonic power and time were 1MHz, 1W, and 8min. Image J was used to perform quantitative calculations based on the bacterial area.

[0056] 1.3 Experimental Results

[0057] 1.3.1 Preparation and characterization of emulsions

[0058] as follows Figure 1 As shown in the figure, A is the state of the emulsion droplets before high-pressure homogenization, and B is the state of the droplets after high-pressure homogenization. It can be seen that after high-pressure homogenization, the emulsion droplets generally become smaller and the particle size is uniform. This shows that high-pressure homogenization can make the emulsion droplets smaller and more uniform. Figure 2 As shown, different numbers of cycles (5 times, 10 times, 20 times) under high-pressure homogenization will not have much effect on the droplet size and uniformity of the emulsion, and it is found that with the increase in the number of cycles, the fluidity of the emulsion is also decreasing, that is, the viscosity of the emulsion is decreasing.

[0059] 1.3.2 Emulsion laser confocal microscopy images

[0060] as follows Figure 3 As shown, the red color is the oil phase labeled with Nile red, and the green color is the water phase labeled with sodium fluorescein. It can be seen that the emulsion is an oil-in-water emulsion.

[0061] 1.3.3 Emulsion stability assessment

[0062] Emulsions with different water-oil ratios were prepared, with water-oil ratios of 9:1, 8:2, and 7:3. The microstructures after day 0 (freshly prepared), 1 day, 3 days, 5 days, and 10 days are shown below. Figure 4 As shown in the figure, as the proportion of the oil phase increases, the emulsion is easy to separate, and conversely, as the proportion of the oil phase decreases, the viscosity of the emulsion decreases. It is also found that the particle size of the emulsion droplets gradually increases with time. The particle size is evaluated and plotted using Image J, as shown below Figure 5 shown.

[0063] 1.3.4 In vivo transdermal effect of ionic liquid emulsion under ultrasound

[0064] like Figure 6 As shown in the figure, the transdermal penetration enhancement effect of FITC-Dextran with a molecular weight of 5000 in medical ultrasound coupling agent (Control), smear emulsion (EM), smear ionic liquid emulsion (IL-EM), ultrasound emulsion (US-EM), and ultrasound ionic liquid emulsion (US-IL-EM) was compared. It can be seen that the transdermal penetration enhancement effect of ionic liquid under ultrasound is the best, both in terms of depth and transdermal strength. And HE staining found that ultrasound conditions will not cause damage to the skin, with a certain degree of safety. The fluorescence intensity was evaluated and plotted by image j. Figure 7 For quantitative analysis of fluorescence intensity, Figure 8-Figure 9 Fluorescence colocalization analysis shows the trend of fluorescence intensity with the change of skin depth. Scale bar: 0.2 mm. It was found that the ionic liquid emulsion has the best effect on the skin depth of FITC-Dextran under ultrasound conditions.

Claims

1. An ionic liquid emulsion, characterized in that: The invention consists of an aqueous phase and an oil phase in a volume ratio of 9-7:1-3. The aqueous phase is an ionic liquid containing organic acid and a total alkaloid extract of Sophora flavescens, and the oil phase is an emulsion containing a transdermal penetration enhancer, a solvent and hydroxypropyl methylcellulose.

2. The ionic liquid emulsion according to claim 1, characterized in that The total alkaloid extract of Sophora flavescens includes matrine, oxymatrine, sophoridine, sophoracarpine or oxysophoracarpine.

3. The ionic liquid emulsion according to claim 1, characterized in that The transdermal penetration enhancer is one or more of lavender essential oil, rosemary essential oil, peppermint essential oil, eucalyptus essential oil, tea tree essential oil, geranium essential oil, frankincense essential oil, myrrh essential oil, carrot seed essential oil, sweet orange essential oil, lemon essential oil, grapefruit essential oil, ylang ylang essential oil, patchouli essential oil, sandalwood essential oil, basil essential oil, fennel essential oil, cinnamon essential oil, clove essential oil, and ginger essential oil.

4. The ionic liquid emulsion according to claim 1, characterized in that The molar ratio of the organic acid to the total alkaloid extract of Sophora flavescens is 2-1:1-2.

5. The ionic liquid emulsion according to claim 1, characterized in that The dosage ratio of the transdermal penetration enhancer, the solvent and the hydroxypropyl methylcellulose is 5-15ml:45-35ml:1.35-1.05g.

6. A method for preparing the ionic liquid emulsion according to any one of claims 1 to 5, characterized in that: The following steps are involved: (1) dissolving hydroxypropyl methylcellulose in a solvent, stirring and mixing, adding a transdermal penetration enhancer, shearing, and homogenizing to obtain an emulsion; (2) dissolving an organic acid and a total alkaloid extract of Sophora flavescens in a solvent for reaction, and removing the solvent after the reaction to obtain an ionic liquid; (3) Dissolving the ionic liquid in the emulsion to obtain the ionic liquid emulsion.

7. The method according to claim 6, characterized in that: In step (1), the stirring and mixing conditions are: stirring at 35-40° C. for 10-15 min; the shearing conditions are: shearing at 8000-10000 rpm for 5-10 min; and the homogenizing conditions are: homogenizing at a homogenizing pressure of 460-500 bar for 5-20 min, and circulating 5-20 times.

8. Use of the ionic liquid emulsion according to any one of claims 1 to 5 in drug penetration and delivery.

9. The use according to claim 8, characterized in that: The ionic liquid emulsion performs drug penetration and delivery under ultrasound, and the ultrasound conditions are: ultrasound frequency is 1-2 MHz, ultrasound power is 0.8-1 W, and ultrasound time is 10-30 min.

10. The use according to claim 8, characterized in that: The penetration depth is 0-1 mm.

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