CN106604738B

CN106604738B - Pharmaceutical composition containing Massa Medicata Fermentata extract for wound treatment

Info

Publication number: CN106604738B
Application number: CN201580008972.8A
Authority: CN
Inventors: 马珍烈; 李正珍; 权慧恩; 金爱英; 金兑洙; 赵元卿
Original assignee: Korea Institute of Oriental Medicine KIOM
Current assignee: Korea Institute of Oriental Medicine KIOM
Priority date: 2014-02-20
Filing date: 2015-02-17
Publication date: 2021-07-13
Anticipated expiration: 2035-02-17
Also published as: KR20150098436A; CN106604738A; WO2015126151A3; WO2015126151A2; KR101574591B1

Abstract

The present invention relates to a pharmaceutical composition, a quasi-drug composition and a health functional food composition for wound treatment, each containing a medicated leaven extract as an effective ingredient, and a method for wound treatment, comprising the step of administering the composition to an individual or applying the composition to the skin of the individual. The composition containing the medicated leaven extract of the present invention induces keratinocyte proliferation to exhibit an excellent wound treatment effect, and thus can be effectively used in drugs for wound treatment including skin external preparations.

Description

Pharmaceutical composition containing Massa Medicata Fermentata extract for wound treatment

Technical Field

The present invention relates to a pharmaceutical composition, a quasi-drug composition and a health functional food composition for wound treatment, each containing a medicated leaven extract as an effective ingredient, and a method for wound treatment comprising administering the composition to a subject or applying the composition to the skin of a subject.

Background

The skin, which plays a role in protecting the body, is the first line of defense in the body against disease. The epidermis provides a barrier against microbial ingress. Thus, in the treatment of wounds, burns, abrasions, and other skin related injuries, the primary goal is to prevent rapid closure of infections and wound healing. Generally, wound healing is a complex process comprising 3 stages: inflammation, proliferation and remodeling. The first stage is coagulation to achieve hemostasis and recruitment of neutrophils to destroy bacteria and necrotic tissue, followed by concomitant recruitment of macrophages. During the second phase, angiogenesis occurs, at which time endothelial cells enter the injured area and, at the same time, fibroblasts enter the injured area, which helps to generate granulation tissue. The formation of granulation tissue enables epidermal regeneration to occur. During the final phase, the production and destruction of collagen is balanced and the healed wound is slowly altered to reach maximum strength. When any of these stages does not operate in an appropriate or timely manner, wound healing is delayed or disrupted. This can cause chronic wounds, and such wounds can be not only serious problems for individuals, but also costly clinical problems. Thus, there is a need for compositions that accelerate the natural bodily processes that regenerate skin and epidermal tissues.

Wound healing refers to the clinical complete closure of the skin, which is a complex process in which many types of cells, such as keratinocytes, fibroblasts, endothelial cells, macrophages, platelets, etc., interact and regulate each other. Each process is regulated by a complex signaling network that is regulated by various growth factors, cytokines, and chemokines.

Wounds must be treated rapidly to reduce the risk of secondary infection. Furthermore, if the inflammatory response persists during wound therapy, the inflammatory phase may not progress to the proliferative phase, thereby delaying normal wound healing. For this reason, a great deal of research has been conducted on the development of wound therapeutics that do not induce inflammatory responses.

On the other hand, medicated leaven, which is a herb used since ancient times, is reported to have various pharmacological activities. The medicated leaven is known to enhance the function of the digestive tract and is effective for abdominal pain caused by postpartum blood stasis.

In addition, as for the skin-related effects of medicated leaven, korean patent application laid-open No. 10-2011-0112231 discloses that the encapsulated fermented extracts of various natural products (such as medicated leaven) have antioxidant, anti-aging, whitening, moisturizing, etc. effects on the skin. However, the wound healing effect on medicated leaven is not known.

Disclosure of Invention

Technical problem

The present inventors paid attention to keratinocytes among cells known to play an important role in wound treatment, and focused on finding natural herbal medicines having wound treatment effects using keratinocytes. As a result, they found that the medicated leaven extract has a wound healing effect due to its activity of inducing keratinocyte proliferation, thereby completing the present invention.

Technical scheme

The present invention aims to provide a pharmaceutical composition for wound treatment containing a medicated leaven extract as an active ingredient.

It is another object of the present invention to provide a quasi-drug composition for wound treatment containing a medicated leaven extract as an effective ingredient.

Still another object of the present invention is to provide a health functional food composition for wound treatment containing the medicated leaven extract as an effective ingredient.

It is still another object of the present invention to provide a method for wound treatment, which comprises administering to a subject a composition comprising a medicated leaven extract as an effective ingredient.

It is still another object of the present invention to provide a method for wound treatment, which comprises applying a composition containing a medicated leaven extract as an effective ingredient to the skin of an individual.

Advantageous effects

The composition containing a medicated leaven extract of the present invention induces keratinocyte proliferation, thereby exhibiting excellent wound treatment effects, and thus, can be effectively used in drugs for wound treatment (e.g., skin external preparations).

Drawings

FIG. 1 shows the results of confirming the effect of the Massa Medicata Fermentata extract (50. mu.g/mL and 100. mu.g/mL) of the present invention on the induction of keratinocyte proliferation;

fig. 2a shows the results of confirming the wound treatment effect of the medicated leaven extract (50 μ g/mL and 100 μ g/mL) of the present invention against keratinocytes with naked eyes, and fig. 2b shows the above results in a graph.

Detailed Description

In one aspect for achieving the above objects, the present invention provides a pharmaceutical composition for wound treatment containing a medicated leaven extract as an effective ingredient.

The medicated leaven of the present invention is a substance conventionally used as herbal medicines and the like. In the present invention, the induction activity of medicated leaven to keratinocyte proliferation and the wound healing effect after wound generation were confirmed, thereby disclosing a novel use for wound treatment.

As used herein, the term "medicated leaven (also referred to as mold, medicated leaven, medicinal yeast) refers to a medicinal material prepared by mixing flour, bran, cocklebur fruit juice, anemarrhena juice, sweet wormwood juice, mashed almonds, red beans, etc. in korean medicine and making into yeast-like followed by fermentation. Medicated leaven acts on the spleen and stomach to strengthen the function of digestive tract, and has the functions of promoting digestion and relaxing stomach. It is known to be effective in treating dyspepsia, chest distress, emesis, diarrhea, abdominal pain due to puerperal blood stasis, etc. As for dermatology, it is known to have an effect on facial blood circulation, but its effect on wound treatment is not known yet. Thus, in the present invention, a novel effect of medicated leaven on wound treatment is revealed.

The term "extract" as used herein refers to a preparation obtained by squeezing a crude drug out of an appropriate leachate, and then evaporating and concentrating the leachate. Although not limited thereto, the extract may be an extract obtained by an extraction treatment, a diluted or concentrated solution of an extract, a dried product obtained by drying an extract, and a partially purified or purified product thereof. The medicated leaven extract can be prepared by general extraction methods, isolation and purification methods known in the art. The extraction method may preferably use boiling water extraction, hot water extraction, cold-dip extraction, reflux condensation extraction, ultrasonic extraction, or the like, and more preferably may use hot water extraction, but is not limited thereto.

In the present invention, the extract may be prepared by extracting the medicated leaven with an extraction solvent or adding a separation solvent to the extract prepared by the extraction with the extraction solvent and separating it. The extraction solvent may be, but not limited to, water, an organic solvent such as an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, and a polar solvent such as ethyl acetate or acetone, a nonpolar solvent such as hexane or dichloromethane, or a mixed solvent thereof may be used. Further, preferably, water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof may be used, and more preferably, water is used. In an exemplary embodiment of the present invention, medicated leaven is added to water, soaked, and subjected to hot water extraction, thereby producing a medicated leaven extract.

All of the medicated leaven extracts of the present invention can be effectively used for wound treatment. Wounds, collectively referred to as injured organisms, are also referred to as wounds. In the exemplary embodiment of the present invention, it was confirmed that the medicated leaven extract of the present invention has excellent activity in inducing the proliferation of keratinocytes, respectively, using keratinocytes known to play an important role in wound healing (fig. 1), so that a wound healing effect can be predicted. Further, in another embodiment of the present invention, based on the confirmation that superior wound closure effect was confirmed by treating with the medicated leaven extract, respectively, after the wound was generated by causing abrasion-damage to the actual keratinocytes (fig. 2a and 2b), it was finally revealed that the medicated leaven extract of the present invention is suitable for wound treatment. Therefore, the wound treatment effect of the present invention can be achieved by inducing keratinocyte proliferation.

On the other hand, the pharmaceutical composition of the present invention can be formulated into various forms, respectively, based on the conventional methods. For example, the pharmaceutical composition may be formulated into oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and the like, and may have been formulated into external preparations, suppositories, and sterile injection solutions for use. However, for its purpose in wound treatment, it may be most preferred to provide the composition of the present invention in the form of a skin external preparation. Specifically, the composition may be used in the form of a skin external preparation such as a liquid preparation, an ointment, a cream, a lotion, a spray, a patch, a gel, an aerosol, and the like.

In addition, it may be prepared to further include pharmaceutically acceptable carriers, for example, carriers known in the art, such as buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, binders, excipients, lubricants, etc., on a per dosage form basis. For example, for external preparations for skin to be topically applied in the injured area, conventional additives such as preservatives, solvents for facilitating penetration of drugs, and for ointments and creams, emollients, and the like may be included, and conventional carriers such as ethanol or oleyl alcohol may be included.

In addition, in the pharmaceutical composition of the present invention, a compound generally used for wound treatment, such as an antibiotic, an anti-inflammatory agent, an auxiliary agent for skin treatment, an anesthetic, an analgesic, and the like, may be further contained. These agents may be slightly changed depending on each dosage form, and when provided in the form of a skin external preparation, these agents are preferably contained, but not limited thereto. Generally, an agent such as an antibiotic, an antibacterial agent, or the like may be provided to the injured area to prevent infection of the injured area or wound deterioration caused by the infection. In addition, local anesthetics, analgesics, and the like may be provided for pain relief, and anti-inflammatory agents such as antihistamines may be provided to prevent excessive inflammatory response during wound treatment. By providing a composition comprising various functional drugs, convenience of use can be improved and an environment conducive to effective wound healing can be created, and thus, the effect of wound treatment can be more improved.

In another aspect, the pharmaceutical composition of the invention is administered in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit/risk ratio suitable for medical treatment, but insufficient to cause side effects. The effective dosage level may be determined according to factors including the health of the patient, the type of wound, the severity, the activity of the drug, the sensitivity of the drug, the method of administration, the duration of administration, the route and rate of excretion, the period of treatment, the drug being co-administered or co-administered, and factors well known in the medical arts.

In another aspect, the present invention provides a quasi-drug composition for wound treatment comprising a medicated leaven extract. The medicated leaven extract and the wound healing effect thereof are the same as described above, and the composition of the present invention may be added to a quasi drug for the purpose of healing or improving a wound.

As used herein, the term "quasi-drug" refers to one of the following: a fiber, rubber article or the like for the purpose of treating, alleviating, treating or preventing a disease in a human or animal; non-tool or non-mechanical types and the like having weak or no direct action on the body; and preparations for sterilization, disinsection and the like for the purpose of preventing infection. Thus, the term refers to items excluding those non-tool, non-mechanical or non-device categories among items used for the purpose of diagnosing, treating, alleviating, treating or preventing diseases of humans or animals, and excluding those non-tool, non-mechanical or device categories among items used for the purpose of pharmaceutically affecting the structure and function of humans or animals.

When the composition of the present invention is used as a quasi-drug additive, the composition may be added directly or in combination with other quasi-drugs or quasi-drug ingredients, and may be used appropriately based on the conventional methods. The mixing amount of the effective ingredient may be appropriately determined depending on the purpose of use.

The quasi-drug composition of the present invention may preferably be a disinfectant cleanser, a body wash, a mouth wash (garglin), a wet wipe, a detergent soap, a hand sanitizer, a humidifier pack, a mask, a paste, or a filter pack, but is not limited thereto.

In still another aspect, the present invention provides a health functional food composition for wound treatment containing the medicated leaven extract as an effective ingredient. The medicated leaven extract and the wound healing effect thereof are the same as described above, and the composition of the present invention can be added to health functional foods for the purpose of healing or improving wounds.

As used herein, the term "health functional food" refers to a food prepared or processed into a dosage form such as tablets, capsules, granules, liquids, pills, etc. using raw materials or ingredients having useful functions to the human body. The health functional food of the present invention can be prepared by a method generally used in the art, and at the time of preparation, can be prepared by adding raw materials and ingredients generally added in the art. In addition, any formulation of health functional food may be used, and there is no limitation as long as the formulation is recognized as health functional food.

There is no limitation on the kind of the health functional food to which the composition of the present invention can be applied. Since medicated leaven is known as an edible herb, it can be administered orally in addition to a skin external preparation, and thus, it can be applied to health functional foods for use.

In yet another aspect, the present invention provides a method of wound treatment comprising administering to a subject a composition comprising a medicated leaven extract as an active ingredient.

In still another aspect, the present invention provides a method of wound treatment, which comprises applying a composition containing a medicated leaven extract as an effective ingredient to the skin of an individual.

The medicated leaven extract of the composition and the composition containing the same as an active ingredient are the same as described above, and the wound healing effect is also the same as described above.

The compositions of the present invention may be used for the treatment of wounds or for promoting wound therapy by administration to a subject. Preferably, when provided in the form of a skin external preparation, the composition may be used to treat wounds by being applied to the skin of an individual, and furthermore, the composition may be used not only in therapeutic methods for treating wounds and local and systemic inflammation associated therewith, but also in various diseases associated with or caused by wounds.

As used herein, the term "individual" refers to all animals, e.g., mammals, including mice, livestock, humans, and the like.

Since the medicated leaven extract of the present invention contains natural products, which have been used as herbal medicines since ancient times, as raw materials, no side effects occur when used as pharmaceutical compositions and the like, and thus, it can be safely included in medicines, quasi drugs, and health functional foods for effective use.

Modes for carrying out the invention

Hereinafter, the structure and effect of the present invention will be described in more detail with reference to the following examples. However, these examples are merely illustrative of the present invention, and the scope of the present invention is not limited to these examples.

Example 1: preparation of medicated leaven extract

The medicated leaven extract was prepared based on the method shown below.

1-1: preparation of medicated leaven extract

50g of pulverized medicated leaven was added to 1L of water and soaked for 1 hour. Then, the solution was subjected to hot water extraction for 4 hours using a Daewoong (teddy bear) extractor (DWP-5000M, bengal, korea). After hot water extraction, the resulting solution was concentrated under reduced pressure, and freeze-dried to obtain 100g of a medicated leaven extract.

1-2: preparation method of arillus longan (Longanae arillus) extract

As a comparative group, arillus longan was used. 50g of minced longan pulp is added to 1L of water and soaked for 1 hour. Then, the solution was subjected to hot water extraction for 4 hours using a Daewoong extractor (DWP-5000M, Renchuan, Korea). After hot water extraction, the resulting solution was concentrated under reduced pressure, and freeze-dried to obtain 100g of a arillus longan extract.

1-3: preparation of Kochia scoparia (Kochia scoparia) extract

As a comparative group, kochia scoparia was used. 50g of pulverized Kochia scoparia was added to 1L of water and soaked for 1 hour. Then, the solution was subjected to hot water extraction for 4 hours using a Daewoong extractor (DWP-5000M, Renchuan, Korea). After hot water extraction, the resulting solution was concentrated under reduced pressure, and freeze-dried to obtain 100g of a Kochia scoparia extract.

1-4: preparation method of Fructus Jujubae (Zizyphi Fructus) extract

As a positive control group, jujube whose wound treatment effect was previously known was used. 50g of crushed Chinese dates were added to 1L of water and soaked for 1 hour. Then, the solution was subjected to hot water extraction for 4 hours using a Daewoong extractor (DWP-5000M, Renchuan, Korea). After hot water extraction, the resulting solution was concentrated under reduced pressure, and freeze-dried to obtain 100g of jujube extract.

Example 2: confirmation of Induction Activity for proliferation of human keratinocytes (HaCat)

In order to confirm the wound healing effect of the medicated leaven extract prepared in example 1, the effect on the proliferation of keratinocytes was previously confirmed.

Specifically, 4.0 × 10⁴cells/mL HaCat cells were transferred to 96-well plates and cultured in DMEM (dartbuck modified eagle medium) containing 10% (v/v) Fetal Bovine Serum (FBS) for 24 hours. The medium was treated with the Massa Medicata Fermentata extracts having concentrations of 50. mu.g/mL and 100. mu.g/mL, respectively, and then further cultured for 24 or 48 hours. For the comparison group, the medium was treated with arillus longan and kochia scoparia extract having a concentration of 50 μ g/mL for comparison, and for the positive control group, the medium was treated with jujube extract having a concentration of 500 μ g/mL for comparison, and for the negative control group, the medium was treated with the same amount of sterile distilled water for dissolving the extract. After 22 hours or 46 hours, the medium was treated with CCK-8 reagent (Dojindo, Japan), and the effect of induction of HaCat cell proliferation was measured after 2 hours using optical density. The results are shown in fig. 1.

As shown in fig. 1, first, when the treatment time was 24 hours and when treated with the medicated leaven extract having concentrations of 50 μ g/mL and 100 μ g/mL, it was confirmed that the induction effect on HaCat cell proliferation was significantly increased compared to the effect of the negative control group (control), and it was confirmed that this was similar to the effects of the longan pulp and the kochia scoparia used as the comparative group and the effect of the jujube extract used as the positive control. Furthermore, when the treatment time was 48 hours, the effect of inducing the proliferation of HaCat cells was also significantly increased in all experimental groups treated with the medicated leaven extract having concentrations of 50 μ g/mL and 100 μ g/mL, and the effect could be found to be more significant than that of 24 hours of treatment.

Based on these results, the effect of wound treatment by inducing keratinocyte proliferation can be predicted, and can be confirmed in more detail in the following experiments.

Example 3: confirmation of wound healing Activity

After example 2, in order to confirm the wound healing effect of the medicated leaven extract, keratinocytes were wounded and wound healing activity was confirmed.

Specifically, 2.5 × 10⁵cells/mL HaCat were transferred to 6-well plates and cultured in DMEM containing 10% (v/v) Fetal Bovine Serum (FBS) for 24 hours, after which "bruise damage" was caused on the HaCat cell monolayer by p200 pipette tips. The "bruised injured" HaCat cell layer was treated with the Massa Medicata Fermentata extract at concentrations of 50. mu.g/mL and 100. mu.g/mL and then further cultured for 24 or 48 hours. For the comparison group, the cell monolayers were treated with the arillus longan and the kochia scoparia extract having a concentration of 50 μ g/mL for comparison, and for the positive control group, the cell monolayers were treated with the jujube extract having a concentration of 150 μ g/mL for comparison, and for the negative control group, the cell monolayers were treated with the same amount of sterile distilled water for dissolving the extract.

Thus, as shown in fig. 2a and 2b, it can be confirmed that: in the experimental groups treated with the medicated leaven extract having the concentrations of 50. mu.g/mL and 100. mu.g/mL for 24 hours and 48 hours after the treatment, the area of abrasion was reduced as the "abrasion damage" was cured, compared to the negative control group. FIG. 2a shows photographs morphologically comparing scratch areas, and FIG. 2b shows a graph of area ratios derived from the above results using the Scion-Image (Scion Corporation, MA) program. Based on these results, it was confirmed that: the medicated leaven extract has significantly higher wound healing activity compared to the effect of the negative control group, which has not only higher levels compared to the arillus longan extract and the kochia scoparia extract used as the experimental comparison group, but also similar levels to the jujube extract used as the positive control group.

In summary, it can be confirmed that: the medicated leaven extract has obvious wound treatment effect.

Claims

1. Use of a medicated leaven extract as an active ingredient in the preparation of a medicament for the treatment of wounds, wherein the extract is extracted by hot water.

2. Use according to claim 1, wherein the wound treatment is achieved by inducing keratinocyte proliferation.

3. The use according to claim 1, wherein the medicament is provided in the form of a skin external preparation.

4. The use according to claim 1, wherein the medicament is a liquid formulation, an ointment, a cream, a spray, a patch, a gel or an aerosol.

5. The use of claim 1, wherein the medicament is a lotion.