TWI586381B - Film with a hydrophobic drug, and its manufacturing method and use - Google Patents

Description

Thin film containing hydrophobic drug, and manufacturing method and use thereof

The present invention relates to a film, and in particular to a film containing a hydrophobic drug, and a method and use for the film.

The tree tea oil (TTO) is extracted from the leaves of the Melaleuca alternifolia , which is native to southern Queensland, Australia to the north of New South Wales. It smells like camphor and looks pale yellow to transparent. It has been reported in the literature that tea tree oil has good antibacterial effect (refer to Clin Microbiol Rev. 2006 Jan; 19(1): 50-62), so it has been widely used in the skin beauty or medical industry. In general, tea tree oil can be applied directly to the skin surface to penetrate deep into the skin. Based on the hydrophobic nature of the essential oil of tea tree, the tissue fluid between the skin surface (epidermal cells) and the deep layer of the skin (the dermis cells) is not compatible with the tea tree oil. This phenomenon will reduce the skin penetration of tea tree oil, which will affect its skin beauty or medical effects.

For the sake of the job, how to solve the problem of low skin permeability of hydrophobic drugs such as tea tree oil is one of the problems actively solved by people and/or operators in the technical field to which the present invention pertains.

It is an object of the present invention to provide a novel film comprising the following composition: a hydrophilic matrix, and a hydrophobic drug. The hydrophilic matrix contains silk fibrin protein (SFP) and polyvinyl alcohol (PVA); the hydrophobic drug is coated in a hydrophilic matrix.

Within the scope of the present invention, there is further provided a method of producing the above film comprising the steps of providing a mixture comprising a solvent, silk fibroin, polyvinyl alcohol, and a hydrophobic drug; and drying the mixture to remove the solvent.

Within the scope of the present invention, the use of the above-mentioned film, which is a patch for preparing a skin disease for treating a bacterial infection, is further proposed.

Another object of the present invention is to provide a novel film comprising the following composition: a hydrophilic matrix, and a hydrophobic drug. The hydrophilic matrix contains silk fibroin and polyvinyl alcohol, and has a plurality of pores formed on the surface thereof; the hydrophobic drug is coated in the hydrophilic matrix.

Within the scope of the present invention, there is also proposed a method of producing the aforementioned film comprising the steps of: providing a mixture comprising sodium chloride, a solvent, silk fibroin, polyvinyl alcohol, and a hydrophobic drug; and drying the mixture to remove Solvent; and infiltrating the mixture with water to dissolve the sodium chloride.

Within the scope of the present invention, the use of the aforementioned film, which is a patch for preparing a skin disease for treating a bacterial infection, is further proposed.

In order to more clearly understand the above and/or other objects, functions and features of the present invention, the preferred embodiments are described in detail below:

The first embodiment of the present invention proposes a film which inhibits the growth of bacteria such as Propionibacterium acnes . Thus, the film of the present embodiment can be used as a patch for treating a skin disease of bacterial infection for attachment to the skin of an individual having such a therapeutic need. Examples of skin diseases can be, but are not limited to, acne. The film of the present embodiment contains at least: a hydrophilic matrix and a hydrophobic drug. The hydrophilic matrix contains silk fibroin and polyvinyl alcohol. The hydrophobic drug is coated in a hydrophilic matrix, and examples thereof may be, but not limited to, tea tree essential oil. However, the weight ratio of silk protein, polyvinyl alcohol, and hydrophobic drug may be 100:100:6 to 22.

When the film of the present embodiment is attached to the skin of an individual, the silk fibroin and the polyvinyl alcohol itself have a hydrophilic property, thereby assisting the mutual dissolution of the tissue liquid between the film and the skin surface and the deep layer thereof. In this way, the hydrophobic drug can be quickly delivered to the deep layers of the skin to provide efficacy for therapeutic purposes. Moreover, at the same time of mutual solubility, silk protein itself has high biocompatibility (refer to Adv Drug Deliv Rev. 2013 Apr; 65(4): 457-70), which can reduce the biological toxicity of the film to individuals, and can also supply skin. The nutrients needed to grow. In addition, polyvinyl alcohol can impart a film shape so that it can be attached to the skin for a long time and is not easily disintegrated.

The method for producing the film of the present embodiment will be described below with reference to FIG. 1 :

First, as in step S11, a mixed solution containing a solvent, silk fibroin, and polyvinyl alcohol is prepared. The solvent can assist the silk protein, the polyvinyl alcohol and the subsequent hydrophobic drug to be uniformly mixed, and examples thereof can be, but not limited to, formic acid.

Next, as in step S12, a hydrophobic drug is added to the mixture to obtain a mixture. In order to save time, this step can be combined with step S11; in other words, the solvent, silk fibroin, polyvinyl alcohol, and hydrophobic drug can be directly mixed into a mixture.

Then, as in step S13, the mixture is dried to remove the solvent. Specifically, this step can be accomplished by vacuum drying.

Finally, as in step S14, the dried product is taken out, and the obtained product is the film of the present embodiment.

A second embodiment of the present invention proposes a film which can inhibit the growth of bacteria such as acne bacteria. Based on this, the film of the present embodiment can be used as a patch for treating a skin disease of bacterial infection and attached to the skin of an individual having such a therapeutic need, and examples of the skin disease can be, but not limited to, acne. The film of the present embodiment contains at least: a hydrophilic matrix and a hydrophobic drug. The hydrophilic matrix contains silk fibroin and polyvinyl alcohol, and the hydrophilic matrix is formed with a plurality of pores on the surface thereof, and the pores may have a pore diameter of 4 to 27 μm. The hydrophobic drug is coated in a hydrophilic matrix, and examples thereof may be, but not limited to, tea tree essential oil. The weight ratio of silk protein, polyvinyl alcohol, and hydrophobic drug may range from 100:100:6 to 22.

When the film of the present embodiment is attached to the skin, the hydrophilic property of the silk fibroin and the polyvinyl alcohol itself is compatible with the interstitial fluid between the film and the skin surface and the deep layer. In this way, the hydrophobic drug can be rapidly delivered to the deep layer of the skin, thereby providing efficacy for therapeutic purposes. Furthermore, the high biocompatibility of silk fibroin in the presence of miscibility reduces the biological toxicity of the film to the individual and also provides the nutrients needed for skin growth. Moreover, polyvinyl alcohol can impart a film formation, so that the film can be attached to the skin for a long time so as not to be easily disintegrated. In addition, the pores of the hydrophilic matrix can increase the permeability of the film, making it less prone to skin hypoxia and necrosis.

The method for producing the film of the present embodiment will be described below with reference to FIG. 2:

First, as in step S21, a mixed solution containing a solvent, silk fibroin, and polyvinyl alcohol is prepared. The solvent can assist silk fibroin, polyvinyl alcohol, and evenly mixed with a subsequent hydrophobic drug, sodium chloride, and examples thereof can be, but not limited to, formic acid.

Next, as in step S22, a hydrophobic drug and sodium chloride are added to the mixture to obtain a mixture. In addition to saving time, this step can be combined with step S21; in detail, sodium chloride, solvent, silk fibroin, polyvinyl alcohol, and a hydrophobic drug can be directly mixed into a mixture.

Then, as in step S23, the mixture is dried to remove the solvent. In general, vacuum drying can be used to achieve this step.

Next, as in step S24, the mixture is dried with water to remove sodium chloride. Prior to this step, sodium chloride is present in the dry mixture as a solid sample, so this step dissolves sodium chloride with water to remove sodium chloride from the dry mixture.

Finally, as in step S25, the infiltrated product is taken out, and the obtained product is the film of the present embodiment.

Embodiments of the present invention are exemplified in the following examples.

Preparation Example 1

0.5 g of purified silk protein, 0.5 g of polyvinyl alcohol (Sigma-Aldrich) was added to 2 ml of formic acid (Sigma-Aldrich) and completely dissolved. 3.125 μl (about 0.003 g) of Australian tea tree essential oil (Administry Agricultural Committee Hualien District Agricultural Improvement Field) was added to the obtained formic acid solution; then, the obtained mixture was poured into a disk having a diameter of 3 cm. The mixture was vacuum dried for about 24 hours. Finally, a non-porous film as shown in Fig. 3A was taken out from the disk body.

Preparation Example 2

The non-porous film of this preparation example (Fig. 3B) was prepared according to the procedure shown in Preparation Example 1, except that the volume of the Australian tea tree essential oil was 6.25 μl (about 0.006 g).

Preparation Example 3

The non-porous film of this preparation example (Fig. 3C) was prepared according to the procedure shown in Preparation Example 1, except that the volume of the Australian tea tree essential oil was 12.5 μl (about 0.011 g).

Preparation 4

The non-porous film of this Preparation Example was prepared according to the procedure shown in Preparation Example 1, except that the volume of the Australian tea tree essential oil was 25 μl (about 0.024 g).

Preparation Example 5

The non-porous film of this preparation example was obtained by the procedure shown in Preparation Example 1, except that the volume of the Australian tea tree essential oil was 50 μl (about 0.048 g).

Preparation Example 6

The non-porous film of this preparation example was obtained by the procedure shown in Preparation Example 1, except that the volume of the Australian tea tree essential oil was 100 μl (about 0.096 g).

Comparative Example 1

The non-porous film of this comparative example (Fig. 3D) was prepared according to the procedure shown in Preparation Example 1, except that the Australian tea tree essential oil was not used.

Preparation Example 7

0.5 g of purified silk protein, 0.5 g of polyvinyl alcohol (Sigma-Aldrich) was added to 2 ml of formic acid (Sigma-Aldrich) and completely dissolved. 3.125 μl (about 0.003 g) of Australian tea tree essential oil (Administry Agricultural Committee Hualien District Agricultural Improvement Field) was added to the obtained formic acid solution; then, the obtained mixture was poured into a disk having a diameter of 3 cm and containing 0.05 g of sodium chloride. . The mixture was vacuum dried for about 24 hours. Add pure water to the dried product in the dish and ultrasonically oscillate it. Finally, a film having a hole as shown in Fig. 4A was taken out from the disk body. As shown in Fig. 5, the film of this preparation example has a pore diameter of about 12.02 ± 15.40 μm.

Preparation Example 8

The apertured film of this preparation example (Fig. 4B) was prepared according to the procedure shown in Preparation Example 7, except that the volume of the Australian tea tree essential oil was 6.25 μl (about 0.006 g). As shown in Fig. 5, the film of this preparation example had a pore diameter of about 8.76 ± 6.11 μm.

Preparation Example 9

The apertured film of this preparation example (Fig. 4C) was prepared according to the procedure shown in Preparation Example 7, except that the volume of the Australian tea tree essential oil was 12.5 μl (about 0.011 g). As shown in Fig. 5, the film of this preparation example has a pore diameter of about 3.19 ± 4.22 μm.

Preparation Example 10

The apertured film of this preparation example was prepared in accordance with the procedure shown in Preparation Example 7, except that the volume of the Australian tea tree essential oil was 25 μl (about 0.024 g).

Preparation Example 11

The apertured film of this preparation example was prepared in accordance with the procedure shown in Preparation Example 7, except that the volume of the Australian tea tree essential oil was 50 μl (about 0.048 g).

Preparation Example 12

The apertured film of this Preparation Example was prepared according to the procedure shown in Preparation Example 7, except that the volume of the Australian tea tree essential oil was 100 μl (about 0.096 g).

Comparative Example 2

The apertured film of this comparative example (Fig. 4D) was prepared according to the procedure shown in Preparation Example 7, except that the Australian tea tree essential oil was not used. As shown in Fig. 5, the film of this preparation example has a pore diameter of about 27.23 ± 15.21 μm.

Analysis Example 1

Fibroblast cells are located in the dermis, which mainly produce the interstitial required for dermis cells and maintain skin elasticity. Therefore, this experiment uses MTT method to analyze whether the membrane is toxic to mouse 3T3 cells. As shown in Fig. 6, the membrane-treated cells of Preparation Examples 1, 2, 7, and 8 all had activity higher than 80%.

Analysis Example 2

When hemorrhoids are formed, the lesions are often accompanied by an inflammatory reaction that causes redness and swelling. When an inflammatory reaction occurs, it activates neutrophils and macrophages to produce free radicals, such as nitric oxide (NO). Excessive nitric oxide may cause tissue hypoxia and necrosis; conversely, nitric oxide may promote tissue repair when appropriate. This test is to simulate the inflammatory reaction in vitro, first stimulated mouse RAW264.7 cells with lipopolysaccharides (LPS), and then treated with a thin film. Since nitric oxide is rapidly oxidized to stable nitrite, this experiment uses Griess reagent (Sigma-Aldrich) to directly know the nitrite concentration measured from the membrane-treated cells, and then obtain the membrane-treated cells. Nitric oxide concentration. As shown in Fig. 7, after comparison with only lipopolysaccharide-stimulated cells (positive control group) and only membrane-treated cells (negative control group), Comparative Examples 1, 2, and Preparation Examples 1 to 3, 7 were found. Up to 9 can promote the production of nitric oxide by lipopolysaccharide-stimulated cells, but the concentration of nitric oxide is between the concentration of nitric oxide produced by the positive control group and the negative control group.

Analysis Example 3

After immersing 0.1 g of the film in 2 ml of reinforced clostridial medium (RCM) for about 24 hours, the soaked medium was taken to the bacterium of Acne bacillus (ATCC 6919) and cultured for 24 hours. The culture solution was diluted with 10 times of the medium which was previously only soaked, and then cultured for 24 hours under an anaerobic environment at 37 °C. Finally, the concentration of the cells in the culture solution was measured to determine the inhibitory effect of the film on the acne bacillus. As shown in Figs. 8 and 9, the films of Comparative Examples 1, 2, and Preparation Examples 1 to 3, 7 to 9 all had an inhibitory effect on acne bacteria. Further, the inhibitory effects of the films of Preparation Examples 1 to 3 and 7 to 9 were all more remarkable than those of the films of Comparative Examples 1 and 2.

Analysis Example 4

This test is to analyze the release of the Australian tea tree oil in the buffer. As shown in FIG. 10, the films of Preparation Examples 1 to 3, 7 to 9 can release the tea tree essential oil in the buffer over time, and the percentage of release of the tea tree essential oil is also found in the film (before release). The weight ratio is reversed.

Analysis Example 5

Forty-eight hours after the injection of acne bacteria into the ears of mice, the commercially available 3M Tegaderm dressings, Comparative Examples 1, 2, and Preparations 1, 2, 7, and 8 were applied to the infected ears. As shown in Figs. 11 and 13, the commercially available dressings, the comparative examples 1, 2, and the films of Preparation Examples 1, 2, 7, and 8 all eliminated the redness of the infected ears on the 9th day after the treatment. However, the reddish phenomenon of the ears coated with the films of Comparative Examples 1, 2, and Preparations 1, 2, 7, and 8 on the 3rd day after the treatment was higher than that of the ears coated with the commercially available dressings on the 3rd day after the treatment. The redness is not obvious.

In order to verify the above phenomenon, the amount of remaining bacteria in the ears of the treated mice was specifically analyzed. As shown in Figs. 12 and 14, it can be seen that the amount of remaining bacteria in the tissues of the ears of the preparation examples 1, 2, 7, and 8 after the first and third days after the treatment is less than that of the commercially available dressings. The amount of bacteria remaining in the tissues after the first and third days after treatment. This result represents that the films of Preparation Examples 1, 2, 7, and 8 have better bacteriostatic effects on mouse ears on the first and third days after treatment than commercially available dressings, which is consistent with the above-mentioned phenomenon of ear redness.

The above is only the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto; therefore, the simple equivalent changes and modifications made by the scope of the present invention and the contents of the description of the invention, All remain within the scope of the invention patent.

Steps S11 to S14, S21 to S25‧‧

Fig. 1 is a flow chart for explaining a method of producing a film according to a first embodiment of the present invention. Fig. 2 is a flow chart for explaining a method of producing a film according to a second embodiment of the present invention. Fig. 3A is a photographic view showing the appearance of the film of Preparation Example 1. Fig. 3B is a photographic view showing the appearance of the film of Preparation Example 2. Fig. 3C is a photographic view showing the appearance of the film of Preparation Example 3. Fig. 3D is a photographic view showing the appearance of the film of Comparative Example 1. 4A is a photographic view showing the appearance of the film of Preparation Example 7. Fig. 4B is a photographic view showing the appearance of the film of Preparation Example 8. 4C is a photographic view showing the appearance of the film of Preparation Example 9. 4D is a photographic view showing the appearance of the film of Comparative Example 2. Figure 5 illustrates the pore size of the film of Comparative Example 2 and Preparation Examples 7 to 9. Figure 6 illustrates the membrane-treated mouse 3T3 cell activities of Comparative Examples 1, 2, and Preparations 1 to 12. Figure 7 is a graph showing the concentration of nitric oxide produced by the RAW264.7 cells of the post-treated mice of Comparative Example 1, 2, Preparations 1 to 3 and 7 to 9 by lipopolysaccharide stimulation. Fig. 8 is a view showing the inhibitory effect of the acne bacillus of the film of Comparative Example 1, and Preparation Examples 1 to 3. Fig. 9 is a view showing the acne bacillus inhibitory effect of the film of Comparative Example 2 and Preparation Examples 7 to 9. Figure 10 is a view showing the release of tea tree essential oil in the films of Preparation Examples 1 to 3 and Preparation Examples 7 to 9 in a buffer. Figure 11 is a view showing the appearance of a mouse ear to which a 3M Tegaderm dressing, a comparative example 2, and a film of Preparation Examples 7 and 8 were applied after the first infection of the acne bacillus. Figure 12 illustrates the amount of bacteria in the ear tissue of the mouse of Figure 11. Fig. 13 is a view showing the appearance of a mouse ear to which a 3M Tegaderm dressing, a comparative example 1, and a film of Preparation Examples 1 and 2 were applied before the infection with acne bacteria. Figure 14 illustrates the amount of bacteria in the ear tissue of the mouse of Figure 13.

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Claims (11)

A film comprising: a hydrophilic matrix comprising silk fibrin protein (SFP) and polyvinyl alcohol (PVA); and a hydrophobic drug coated in the hydrophilic matrix; The weight ratio of the silk protein, the polyvinyl alcohol, and the hydrophobic drug is 100:100:6 to 22. The film of claim 1, wherein the hydrophobic drug is tree tea oil (TTO). The method for producing a film according to any one of claims 1 to 2, further comprising: providing a mixture comprising a solvent, the silk protein, the polyvinyl alcohol, and the hydrophobic drug; and drying The mixture is removed to remove the solvent. The use of a film according to any one of claims 1 to 2 for the preparation of a patch for treating a skin disease of a bacterial infection. The use of claim 4, wherein the skin disorder is acne. A film comprising: a hydrophilic matrix comprising silk fibroin and polyvinyl alcohol, and having a plurality of pores formed on the surface thereof; and a hydrophobic drug coated in the hydrophilic matrix; wherein the silk fibroin The weight ratio of the polyvinyl alcohol to the hydrophobic drug is from 100:100:6 to 22. The film of claim 6, wherein the pores have a pore diameter of 4 to 27 μm. The film of claim 6, wherein the hydrophobic drug is tea tree oil. The method for producing a film according to any one of claims 6 to 8, further comprising: providing a mixture comprising sodium chloride, a solvent, the silk protein, the polyvinyl alcohol, and the hydrophobicity a drug; drying the mixture to remove the solvent; and soaking the dried mixture with water to dissolve the sodium chloride. The use of a film according to any one of claims 6 to 8 for the preparation of a patch for treating a skin disease of a bacterial infection. The use of claim 10, wherein the skin disorder is acne.