CN106821981A - A kind of infrared light increases activity RNA（RNAa）Sequence enters skin and produces the medication of bioactivity - Google Patents

Abstract

The invention relates to a drug delivery method for increasing the sequence of activating RNA (RNAa) into the skin by infrared light and producing biological activity, comprising the following steps: (1) dissolving liposomes in optimized culture medium and standing; (2) Dissolve the activating RNA sequence in the optimized culture medium and let stand; (3) Mix the solutions of step (1) and step (2) and let stand; (4) Dissolve the solution of step (3) in the carrier solution (5) For the site where the skin needs to be administered, after irradiating with a certain dose of infrared light, apply the solution described in step (4) evenly on the irradiated site; the wavelength range of the infrared light is 1500nm-2000nm. The invention uses infrared light to irradiate the skin to increase the amount of activating RNA sequences entering the skin, and has biological activity on the skin, laying the foundation for the clinical use of activating RNA sequences to treat skin diseases, allowing patients to receive more effective treatment.

Description

An infrared light increases the entry of activating RNA (RNAa) sequences into the skin and produces biological active drug delivery method

technical field

The invention relates to a drug delivery method that uses infrared light of a certain waveband to increase the entry of activating RNA sequences into the skin and exert biological effects in the skin.

Background technique

Some activating RNAs have been shown to be useful in the treatment of skin diseases such as skin melanoma, congenital pachyonychia, skin inflammation, skin cancer, etc. However, due to the complex physiological environment in the gastrointestinal tract, the RNA sequence will eventually be degraded, and it is difficult for biological macromolecules to pass through the gastrointestinal tract membrane. The intravenous administration method will cause the patient a strong sense of pain, and the RNA sequence will be metabolized or excreted before reaching the skin. None of these traditional methods of drug delivery had a significant effect at one time. At present, the most commonly used skin drug delivery method is to directly give the activating RNA sequence to the skin, so as to regulate the gene expression of the skin tissue in situ, thereby achieving a therapeutic effect.

Due to the tight structure of the stratum corneum and the epidermis, it is an important factor for the greatly reduced permeability of the drug. Therefore, it is particularly important to improve the permeability of the skin so that the activating RNA sequence can enter the skin and have a therapeutic effect. At present, the methods of improving skin permeability that have been compared and studied are mainly divided into chemical enhancers and physical enhancers. Chemical enhancers use nanomaterials, liposomes, etc. to increase the permeability of the activating RNA sequence to the skin. This type of enhancer has high requirements on the length and structure of the activating RNA sequence, and at the same time, the preparation process is cumbersome and complicated. The more popular physical enhancers include microneedle drug delivery, electrical drug delivery and other methods. However, microneedle administration and electrical administration can cause great pain to patients.

So far, in the research on the use of laser to enhance the permeability of activating RNA sequences to the skin, there is no report that the use of laser can effectively enhance the biological activity of activating RNA sequences in the skin. These biological activities can only be proved by methods such as the detection of the content of the target protein and the detection of the fluorescence produced by the target protein. Therefore, it is of great clinical significance to invent a method that can effectively and significantly improve the biological activity of exogenous activating RNA sequences in the skin.

Contents of the invention

The purpose of the present invention is to overcome the shortcomings of the above-mentioned prior art. The inventors found that after the skin is irradiated with infrared light, the biological effect of the activating RNA sequence on the skin is significantly increased. This effect is proved by the detection of the content of the target protein and the detection of the fluorescence produced by the target protein. Therefore, based on this new discovery, the present invention breaks through the difficulties of the prior art and finds a method that can improve the biological effect of the activating RNA sequence on the skin.

The specific technical solution of the present invention is a drug delivery method in which infrared light increases the sequence of activating RNA (RNAa) into the skin and produces biological activity, comprising the following steps:

(1) Dissolve liposomes in optimized culture medium and let stand;

(2) Dissolve the activating RNA sequence in the optimized culture medium and let stand;

(3) Mix the solutions of step (1) and step (2) and let stand;

(4) Dissolving the solution of step (3) in the carrier solution;

(5) For the site where the skin needs to be administered, after irradiating with a certain dose of infrared light, apply the solution described in step (4) evenly on the irradiated site; the wavelength range of the infrared light is 1500nm-2000nm.

Further, the ratio of the liposome to the optimized culture medium can be adjusted according to the administration effect.

Further, the ratio of the activating RNA sequence to the optimized culture medium can be adjusted according to the administration effect.

Further, the ratio of the liposome to the activating RNA sequence can be adjusted according to the administration effect.

Further, all the resting times can be adjusted according to the administration effect.

The invention uses infrared light to irradiate the skin, increases the amount of activating RNA sequence entering the skin, and exerts biological activity on the skin, and establishes a basis for clinically using the activating RNA sequence to treat skin diseases, enabling patients to receive more effective treatment.

Description of drawings

Fig. 1 Fluorescent three-dimensional view of fluorescently labeled activating RNA sequences entering the skin.

Figure 2 Fluorescent projection of fluorescently labeled activating RNA sequences entering the skin.

detailed description

The present invention will be further illustrated by specific description below.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

It is determined that infrared light can increase the biological activity of activating RNA sequences in the skin, and a novel skin-activating RNA sequence drug delivery method has been established.

According to the present invention, male C57 mice with a weight ranging from 15-25 g are used. After the administration of the activating RNA sequence is completed, the mice are raised in the animal room under the conditions of 22-24° C., 12 hours of light/dark Circulation, and free access to food and water.

Mouse skin using fluorescently labeled RNA-seq was imaged using confocal laser microscopy over a 24-hour period.

Within 24 hours, detect the mouse skin using the RNA sequence of the target protein for analysis of the target protein content.

Within 24 hours, the mouse skin was damaged by ultraviolet light, and the biological effect of using RNA sequence was detected.

Storage of skin tissue: After the skin was taken out, an appropriate amount of tissue was taken out and soaked in 4% paraformaldehyde for making paraffin sections. The remaining tissue was wrapped in aluminum foil, frozen with liquid nitrogen, and then transferred to a -80°C refrigerator for long-term storage.

Quantify the target protein content in the skin.

Statistical analysis: All data are given in the form of mean ± standard deviation, and the data are evaluated by one-way analysis of variance, and a P value less than 0.05 is considered statistically significant.

The results of the mouse experiments are as follows:

Figure 1: Three fluorescent views of fluorescently labeled activating RNA sequences entering the skin, skin treated with infrared light, more fluorescently labeled activating RNA sequences entering.

Figure 2: Fluorescent projection of fluorescently labeled activating RNA sequences entering the skin, more fluorescently labeled activating RNA sequences entering the skin treated with infrared light.

From the above experiments, it can be concluded that after the skin is irradiated with infrared light, the increase of activating RNA sequences has biological effects on the skin. Therefore, more effective skin disease treatment can be carried out using this method.

Based on this discovery, the present invention establishes a drug delivery method for increasing the sequence of activating RNA (RNAa) into the skin by infrared light and producing biological activity, the method comprising the following steps:

(1) Dissolve liposomes in optimized culture medium and let stand for a certain period of time

(2) Dissolving the activating RNA sequence in the optimized culture medium and standing for a certain period of time;

(3) Mix the solution of step (1) and step (2) and let it stand for a certain period of time;

(4) Dissolving the solution of step (3) in the carrier solution;

(5) For the site where the skin needs to be administered, after irradiating with a certain dose of infrared light, apply the solution described in step (4) evenly on the irradiated site; the wavelength range of the infrared light is 1500nm-2000nm.

In the preparation method of the activating RNA sequence of the present invention, the ratio of the activating RNA sequence to the optimized culture medium can be adjusted according to the administration effect.

In the preparation method of the activating RNA sequence of the present invention, the ratio of the liposome to the optimized culture medium can be adjusted according to the administration effect.

In the preparation method of the activating RNA sequence of the present invention, the ratio of the activating RNA sequence to the liposome can be adjusted according to the administration effect.

The invention is based on irradiating the skin with infrared light of a certain wave band, thereby significantly improving the mixture of liposome and activating RNA RNA sequence to enter the skin and exert biological effects on the skin. This effect is proved by methods such as detection of the content of the target protein and detection of the fluorescence produced by the target protein. This approach could significantly increase the efficacy of activating RNAs for skin diseases.

Those skilled in the art will appreciate that, although specific embodiments of the invention have been described herein for purposes of illustration, various modifications can be made thereto without departing from the spirit and scope of the invention. Therefore, the detailed description and examples of the present invention should not be considered as limiting the scope of the present invention. The invention is limited only by the appended claims. All documents cited in this application are hereby incorporated by reference in their entirety.

Claims (1)

1. A drug delivery method in which infrared light increases the sequence of activating RNA (RNAa) into the skin and produces biological activity, comprising the following steps: (1) Dissolve liposomes in optimized culture medium and let stand; (2) Dissolve the activating RNA sequence in the optimized culture medium and let stand; (3) Mix the solutions of step (1) and step (2) and let stand; (4) Dissolving the solution of step (3) in the carrier solution; (5) For the site where the skin needs to be administered, after irradiating with a certain dose of infrared light, apply the solution described in step (4) evenly on the irradiated site; the wavelength range of the infrared light is 1500nm-2000nm.