TWI715838B - Uses of a composition in the manufacture of medicaments for promoting hair growth, preventing hair loss or promoting the proliferation of hair follicle stem cells in a subject - Google Patents

Abstract

The present invention relates to the use of a composition in the manufacture of medicaments for promoting hair growth, preventing hair loss or promoting the proliferation of hair follicle stem cells in a subject, wherein the composition comprises Deriant, a sodium deoxyribonucleate, as the main active ingredient.

Description

Use of a composition for preparing a medicament for promoting individual hair growth, preventing hair loss or promoting the production of individual hair follicle stem cells

The present invention relates to the use of a composition for preparing a medicament for promoting individual hair growth, preventing hair loss or promoting the production of individual hair follicle stem cells, wherein the composition comprises an effective dose of Derinat, a deoxyribonucleic acid Sodium salt, as the main active ingredient.

Hair follicles are important accessory organs of the skin used to grow hair, and are composed of various types of cells including hair follicle stem cells. The cycle experienced by hair follicles includes the anagen stage, the catagen stage, and the telogen stage, and the three stages are repeated continuously. The activation period is characterized by the activation of hair follicle stem cells. During this period, the hair follicle stem cells continue to proliferate and grow toward the subcutaneous tissue. In the middle of the activation period, the hair follicle stem cells stop dividing and enter a hibernation state, and the work of hair growth is handed over to the outer root sheath, inner root sheath, and matrix differentiated from the hair follicle stem cells. Matrix cells are responsible, and these cells finally differentiate into a structure with hair scales to form a hair shaft visible to the naked eye.

Abnormal hair follicle cycles often lead to severe hair failure or hair loss. Baldness is one of the most common conditions. Baldness affects the individual's body and mind, regardless of gender, and the weakening of self-esteem affects many aspects of their lives, which is disturbing.

At present, there is no product on the market that can effectively increase hair growth. At most, it can only increase the stability of the hair structure, reduce the chance of hair loss, or inhibit 5α-reductase (5α-reductase) to make dihydrotestosterone (DHT). ) The output is reduced to achieve the purpose of curing baldness, but the latter has the adverse side effects of sexual dysfunction. Therefore, it is necessary to provide an effective and non-toxic composition or method with relatively no adverse side effects to promote hair growth or prevent hair loss.

The Derinat composition is a natural deoxyribonucleic acid (DNA) sodium salt composition extracted from fish eggs (mostly trout or salmon), which is composed of more than 90% DNA sodium salt The composition is about 100~300 base pairs in size. The present invention discloses that the high skin cell dose (1.5 milligrams/ml (mg/ml)) of this sodium salt is not toxic to human skin keratinocytes (KC) and human dermal fibroblasts (HDF) . The Daili sodium salt of the present invention can reduce the concentration of calcium ions in skin cells, and achieve the effects of protecting skin cells and reducing the production of reactive oxygen species (ROS) in skin cells. Furthermore, experiments with nude mice found that the back of the nude mice was exposed to the composition containing an effective dose of Dai Li sodium salt for two hours every day, and after a week, the hair on the back of the nude mice increased; then the nude mice were sacrificed to take the back In the skin, hematoxylin-eosin staining (H&E Stain) found that the hair follicles were hyperplasia, and more hair follicles showed anagen stage. Since the proliferation of hair follicles is related to the activation of stem cells, immunohistochemistry (IHC) staining was used to observe the molecular markers CD49f and Oct-3/4 that the two stem cells would exhibit. The results showed that after treatment with the Daili sodium salt , There are more hair follicle stem cells. Not only that, the Daili sodium salt can also reduce the generation of oxidative free radicals in hair follicle cells, and has the function of protecting skin and hair follicle cells. Based on the above experimental results, Daili sodium salt is not only non-toxic to skin cells, but can further promote hair follicle proliferation and activate hair follicle stem cells; in addition, it can also reduce the production of oxidative free radicals in skin tissues, thereby achieving the effect of protecting skin cells and hair follicle cells .

The present invention relates to the use of a composition for preparing a medicament for promoting individual hair growth or preventing hair loss, wherein the composition contains an effective dose of Derinat.

In one embodiment, the composition contains 10-1500 micrograms per milliliter (μg/mL) of Dairyi sodium salt.

In one embodiment, the composition contains 10-150 micrograms per milliliter (μg/mL) of Dairyi sodium salt.

In one embodiment, the composition refers to a pharmaceutical composition applied through the skin.

When the composition is prepared into a medicine, the unit dose of Dai Li sodium salt contained in each unit of the medicine is 10-1500 micrograms (μg), more preferably 10-150 micrograms (μg).

In one embodiment, the subject includes a mammal that has no hair growth, hair loss, or is at risk of hair loss or hair loss.

In one embodiment, the hair is located on the scalp, face, beard, head, trunk, limbs, pubic area, upper lip, eyebrows or eyelids.

The present invention also relates to the use of a composition for preparing a medicament for promoting the production of individual hair follicle stem cells, wherein the composition contains an effective dose of Dai Li sodium salt.

In one embodiment, the composition contains 10-1500 micrograms per milliliter (μg/mL) of Dairyi sodium salt.

In one embodiment, the composition contains 10-150 micrograms per milliliter (μg/mL) of Dairyi sodium salt.

In one embodiment, the composition refers to a pharmaceutical composition applied through the skin.

When the composition is prepared into a medicine, the unit dose of Dai Li sodium salt contained in each unit of the medicine is 10-1500 micrograms (μg), more preferably 10-150 micrograms (μg).

In one embodiment, the individual includes a mammal with non-growth hair, hair loss, or at risk of non-hair growth or hair loss.

In one embodiment, the hair is located on the scalp, face, beard, head, trunk, limbs, pubic area, upper lip, eyebrows or eyelids.

The present invention also provides a method for promoting hair growth or preventing hair loss in an individual, which comprises locally administering to the individual a composition containing an effective dose of Dai Li sodium salt.

In one embodiment, the composition contains 10-1500 micrograms per milliliter (μg/mL) of Dairyi sodium salt.

In one embodiment, the composition contains 10-150 micrograms per milliliter (μg/mL) of Dairyi sodium salt.

In one embodiment, the composition refers to a pharmaceutical composition applied through the skin.

In one embodiment, the individual includes a mammal with non-growth hair, hair loss, or at risk of non-hair growth or hair loss.

In one embodiment, the hair is located on the scalp, face, beard, head, trunk, limbs, pubic area, upper lip, eyebrows or eyelids.

The present invention further provides a method for promoting the production of hair follicle stem cells in an individual, which comprises locally administering a composition containing an effective dose of Dai Li sodium to the individual.

In one embodiment, the composition contains 10-1500 micrograms per milliliter (μg/mL) of Dairyi sodium salt.

In one embodiment, the composition contains 10-150 micrograms per milliliter (μg/mL) of Dairyi sodium salt.

In one embodiment, the composition refers to a pharmaceutical composition applied through the skin.

In one embodiment, the individual includes a mammal with non-growth hair, hair loss, or at risk of non-hair growth or hair loss.

In one embodiment, the hair is located on the scalp, face, beard, head, trunk, limbs, pubic area, upper lip, eyebrows or eyelids.

The composition referred to in the present invention includes a composition for topical and regional effects. The term "topical" as used herein refers to the use of the composition described herein incorporated into a suitable pharmaceutical carrier and applied to areas of thinning or alopecia to achieve a topical effect. Therefore, such topical compositions include those pharmaceutical forms in which the compound is applied externally by direct contact with the skin surface to be treated. Conventional pharmaceutical forms used for this purpose include ointments, wipes, creams, shampoos, lotions, pastes, gels, sprays, aerosols, etc., and can be in patches or according to the body part to be treated Application in the form of dipping dressing. The term "ointment" includes preparations (including creams) having an oily base, a water-soluble base, and an emulsion base such as petrolatum, lanolin, polyethylene glycols, and mixtures of these combinations.

For topical application of the eyebrows or eyelids, the drug can be formulated in an aqueous solution, cream, ointment or oil by adding pharmacologically acceptable buffers and salts. Such pharmaceutical compositions may or may not contain preservatives (e.g., benzalkonium chloride, chlorhexidine, chlorobutanol, p-hydroxybenzoic acid) and phenyl mercury salts (e.g., nitrate, chloride, acetate, and borate). ) Or antioxidants and additives (such as EDTA, sorbitol, boric acid, etc.). In addition, especially the aqueous solution may contain a viscosity-increasing agent such as polysaccharides (such as methylcellulose), mucopolysaccharides (such as hyaluronic acid and chondroitin sulfate) or polyols (such as polyvinyl alcohol). According to the substance that forms an in situ gel (in situ gel, refers to a preparation that can undergo a phase transition at the application site after being administered in a solution state, from a liquid state to a semi-solid gel state), and different slow-release gels can be used Glue and matrix, as well as soluble and insoluble ophthalmic inserts. In actual situations, different amounts of drugs and different dosages can be used as preparations.

For topical application to the skin and scalp, it may be advantageous to use ointments, creams, wipes or patches as active ingredient carriers to formulate pharmaceutical compositions. In addition, these pharmaceutical compositions may or may not contain preservatives. Such preservatives include the above-mentioned pharmaceutical compositions as well as methyl, propyl- or butyl-p-hydroxybenzoic acid, betaine, chlorhexidine, benzalkonium chloride and the like. The pharmaceutical composition can also be delivered with sustained release through different bases. To achieve the daily dosage of the pharmaceutical composition, the composition can be administered one or more times a day in the presence or absence of antioxidants.

The condition of hair loss caused by the lack of hair growth or hair loss according to the present invention can be hair loss (for example, androgenic alopecia, alopecia areata, etc.), hereditary baldness, scars, burns, radiotherapy, chemotherapy, and disease-related Caused by hair loss, accidental injury, hair follicle injury, surgical trauma, incision injury, or trauma to the receiving site of the skin graft.

The present invention can be applied to all mammalian species, including both humans and animals. In humans, the composition of the present invention can be applied to the scalp, face, beard, head, trunk, limbs, pubic area, upper lip, eyebrows or eyelids. In animals raised for obtaining fur, the composition of the present invention can be applied to the entire body surface to improve the entire fur for commercial reasons. For aesthetic reasons, the composition of the present invention can be applied to animals, for example, to the skin of dogs or cats that have a certain degree of baldness due to mite disease or have bald spots due to other diseases.

Unless the context clearly indicates otherwise, the singular forms "a" and "the" used in this specification and the scope of the patent application include plural references. For example, reference to "a derivative" includes a plurality of such derivatives, and reference to "an entity" includes reference to one or more individuals, etc.

Also, unless otherwise stated, the use of "or" means "and/or". Similarly, "include" and "include" are used interchangeably and are not intended to be limiting.

It should be further understood that when the description of each embodiment uses the term "comprising", those familiar with the art should understand that in some specific cases, or may use the language "basically composed of" or "by. ..... Composition" to describe the embodiment.

Unless otherwise defined, all technical and scientific terms used in this article have the same meaning as commonly understood by those skilled in the technical field to which this disclosure belongs. Although methods and materials similar or equivalent to the methods and materials described herein can be used to implement the disclosed methods and compositions, only exemplary methods, devices, and materials are described herein.

The public cases discussed above and in the entire article are only provided for the disclosure content before the filing date of this application. In any respect, this article should not be construed as an admission that the inventor has no right to prioritize the disclosure due to the previous disclosure.

Figure 1 is a series of studies on the toxicity of Dali sodium salt to human skin cells. The results show that the high concentration of Dali sodium salt is 1.5 mg/ml (mg/ml) in skin keratinocytes (KC) and skin fibroblasts (HDF). After the treatment, the cells did not die significantly.

Figure 2 is the treatment of only low concentrations of 15 micrograms/ml (μg/mL) and 150 micrograms/ml (μg/mL) Daili sodium in skin keratinocytes (KC) and skin fibroblasts (HDF) respectively, namely It can inhibit the motivation of calcium ions ([Ca ²⁺ ] _i ).

Figure 3 is to explore the effect of Dai Li sodium salt on the back hair growth of nude mice. It is maintained for one week, and the back of nude mice is exposed to 14 micrograms/ml (μg/mL) or 60 micrograms/ml (μg/mL) Dai Li sodium every day. Within hours, it was found that the back hairs of the nude mice in the Daili sodium salt group increased.

Figure 4 shows that the back skin of nude mice was sacrificed and stained with hematoxylin-eosin (H&E Stain). It was found that the hair follicles in the Daili sodium salt group were proliferated and more hair follicles showed an active period.

Figure 5: The proliferation of hair follicles is related to the activation of hair follicle stem cells. Therefore, immunohistochemistry (IHC) staining was used to observe the molecular markers CD49f and Oct-3/4 that the two hair follicle stem cells would exhibit. The results showed that Daili sodium salt After treatment, there are more hair follicle stem cells.

Fig. 6A is the detection of the expression of reactive oxygen species (ROS) in skin tissue or hair follicle cells by biopsy staining. Figure 6A shows that Dai Li sodium salt can reduce oxidative free radicals in skin cells.

Fig. 6B shows the expression of reactive oxygen species (ROS) in skin tissue or hair follicle cells by biopsy staining. Figure 6B shows that Daili sodium salt can reduce oxidative free radicals in hair follicle cells. The dotted line indicates the range of hair follicles, and the arrow points to the hair follicle cells.

The following examples are not for limiting purposes, but only for presenting various aspects of this invention.

Example 1: Discussion on the toxicity of Dai Li sodium salt to human skin cells

Daili sodium salt was applied to human skin keratinocytes and skin fibroblasts to test its toxicity to human skin cells. The results showed that there was no obvious death of skin keratinocytes (KC) and skin fibroblasts (HDF) under the treatment of high concentration of 1.5 mg/ml (mg/ml) Dai Li sodium salt (Figure 1).

Example 2: Test of Dai Li sodium salt for inhibiting changes in intracellular calcium ion concentration

It is known that when the concentration of calcium ions in skin cells increases, excessive calcium ions will affect mitochondria, causing mitochondria to produce reactive oxygen species (ROS), which in turn causes damage to skin cells. In order to observe the effect of Derinat on the activity of calcium ion channels in skin cells and intracellular calcium ion concentration, the first generation of human skin keratinocytes and skin fibroblasts were treated with Derinat for 24 hours, and then calcium ions were used. Dye (Fluo-4-AM) to detect changes in intracellular calcium ion concentration. When observing the effect of Derinat on calcium ion channels, due to the characteristics of calcium ion channels and more accurate detection of calcium ion channels, ATP is used to stimulate cells to open calcium ion channels when detecting the dynamic changes of intracellular calcium ions. First, use ATP in a calcium ion-free buffer solution to stimulate the opening of calcium ion channels, then wash away the ATP, then change to a buffer solution containing 2 millimolar (mM) calcium ions, then detect the curve height, and do Quantification of height area. Figure 2 shows that compared with the control group not administered Derinat and the group administered DMSO, low concentrations of 15 micrograms/ml (μg/mL) and 150 micrograms/ml (μg/mL) of Dai Li sodium can inhibit The calcium ion channel activity of skin keratinocytes and skin fibroblasts reduces the intracellular calcium ion concentration to achieve the effect of protecting skin cells.

Example 3: Effect of Dai Li sodium salt on hair growth of nude mice

In order to explore the effect of Dai Li sodium salt on the hair growth of nude mice, 14 micrograms/ml (μg/mL) (ie 3.5 micrograms/cm² (μg/cm ² )) or 60 micrograms/ml (μg/mL) (ie 14 micrograms/cm² (μg/cm ² )) of Daili sodium salt was applied to the back of nude mice daily for two hours. After a week of testing, it was found that the hair on the back of the nude mice in the Daili sodium salt group There is an increase (Figure 3). Since 14 micrograms per milliliter (μg/mL) of Daili sodium salt is effective, follow-up experiments will be based on this dose. After the nude mice were sacrificed, the back skin was removed and sectioned with hematoxylin-eosin staining. It was found that the hair follicles in the Daili sodium salt group had hyperplasia, and more hair follicles showed an anagen stage (Figure 4). Since the proliferation of hair follicles is related to the activation of stem cells, tissue immunostaining was used to observe the molecular markers CD49f and Oct-3/4 that the two hair follicle stem cells would exhibit. The results showed that after treatment with Dai Li sodium salt, the molecular markers CD49f and Oct- 3/4 of the expression increased, indicating that more hair follicle stem cells were produced (Figure 5).

Example 4: The effect of Dai Li sodium salt on the generation of oxidative free radicals in skin tissues

In order to further understand the effect of Dai Li sodium salt on the generation of oxidative free radicals in skin tissues, biopsy staining was used to detect the expression of oxidative free radicals in skin cells and hair follicle cells. It was also found that whether it is for skin cells or hair follicle cells, wear Sodium salt can reduce the production of oxidative free radicals, and then achieve the effect of protecting skin cells and hair follicle cells (Figure 6A and Figure 6B).

In order for those with ordinary knowledge in the technical field of this invention to understand the method of making and using this technique, this invention has been described and exemplified in sufficient detail. However, various variations, modifications or improvements should be considered It is tantamount to the spirit and scope of this invention.

Those with ordinary knowledge in the technical field of the present invention can easily understand and achieve the objectives of the present invention, and obtain the results and advantages mentioned earlier. The cells, animals, materials, and the processes and methods for producing them used in the present invention represent the best embodiments, are exemplary in nature, and are not intended to limit the scope of the present invention. Those skilled in the art and the modifications or other uses that will be produced when making or using this technique are all within the spirit of the present invention and are defined by the scope of rights.

Claims (10)

The use of a composition for preparing a medicament for promoting individual hair growth or preventing hair loss, wherein the composition contains an effective dose of Derinat. Such as the use of item 1 in the scope of the patent application, wherein the composition contains an effective dose of the Daili sodium salt of 10-1500 micrograms/ml. Such as the use of item 1 in the scope of the patent application, wherein the composition contains an effective dose of the Daili sodium salt of 10-150 micrograms/ml. Such as the use of item 1 in the scope of patent application, wherein the composition refers to a pharmaceutical composition for transdermal administration. Such as the application of the first item of the scope of patent application, where the hair is located on the scalp, face, beard, head, torso, limbs, pubic area, upper lip, eyebrows or eyelids. The use of a composition for preparing a medicament for promoting the production of individual hair follicle stem cells, wherein the composition contains an effective dose of Daili sodium salt. Such as the use of item 6 of the scope of patent application, wherein the composition contains an effective dose of the Daili sodium salt of 10-1500 micrograms/ml. Such as the use of item 6 in the scope of patent application, wherein the composition contains an effective dose of the Daili sodium salt of 10-150 micrograms/ml. For example, the use of item 6 in the scope of patent application, wherein the composition refers to a pharmaceutical composition for transdermal administration. Such as the use of item 6 of the scope of patent application, where the hair is located on the scalp, face, beard, head, trunk, limbs, pubic area, upper lip, eyebrows or eyelids.

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