CN110755472A - Soybean fermentation composition for skin wound healing and preparation method thereof - Google Patents
Soybean fermentation composition for skin wound healing and preparation method thereof Download PDFInfo
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- CN110755472A CN110755472A CN201910907899.8A CN201910907899A CN110755472A CN 110755472 A CN110755472 A CN 110755472A CN 201910907899 A CN201910907899 A CN 201910907899A CN 110755472 A CN110755472 A CN 110755472A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
Abstract
The invention provides a pharmaceutical composition for healing skin wounds, in particular to a soybean fermentation-induced composition which is a soybean fermentation-induced fermentation broth prepared by co-culturing soybean liquid and more than one microbial probiotic group, wherein the microbial probiotic group is a group consisting of microorganisms of bifidobacterium, lactobacillus, bacillus, lactobacillus and streptococcus. The invention can effectively promote the healing of skin wounds and solve the problems of chronic wound healing and difficult wound healing.
Description
Technical Field
The invention provides a soybean fermentation composition for skin wound healing and a preparation method thereof.
Background
The main functions of the skin include protection, excretion, secretion, absorption, thermal regulation, accumulation, sensory cognition and regulation of immune processes, so that when there is a wound on the skin, such as a wound caused by a general wound or a decubitus ulcer, the skin is adversely affected and fails to perform its normal intended function.
In addition, it is known that the wounds of diabetes are not easy to heal, and patients are easy to have dry skin and easy to crack due to autonomic neuropathy; the patients are also easy to have the end blood vessels blocked easily and the circulation is poor due to the vascular diseases, so that the skin is easy to shrink.
The product of soybean fermentation using microorganisms can also be used as an active oxygen resistant composition, agent, food, cosmetic and pharmaceutical (for example, Japanese patent No. 4139132). Certain specific soy extracts are useful in cosmetics or pharmaceuticals for the treatment of certain skin disorders, for example, soy extracts containing sphingomyelin and phospholipids in specified ratios are disclosed as being useful in cosmetics for the treatment of dry skin (U.S. patent publication No. US 2002/0009509A 1). However, the extracts are obtained by crushing mature whole soybeans or oil-free soybean meal with water to produce a crude extract, followed by treatment with an aliphatic hydrocarbon and an aliphatic ketone.
Disclosure of Invention
Therefore, the present invention provides a new preparation method of a pharmaceutical composition for healing skin wounds, which can effectively promote the healing of skin wounds and solve the problems of chronic wound healing and difficult wound healing.
The composition is a soybean fermentation-promoting composition, which is prepared by co-culturing soybean liquid and more than one microbial probiotic group, wherein the microbial probiotic group is a group consisting of microorganisms of bifidobacterium, lactobacillus, bacillus, lactobacillus and streptococcus.
In one embodiment of the present invention, the microorganism belonging to the genus Bifidobacterium is Bifidobacterium (Bifidobacterium bifidum) or Bifidobacterium longum (Bifidobacterium longum); the microorganism belonging to the genus Bacillus is Bacillus coagulans (Bacillus coagulans); the microorganism of the genus Lactobacillus is selected from the group consisting of Lactobacillus Acidophilus (Lactobacillus Acidophilus), Lactobacillus casei (Lactobacillus casei), Lactobacillus delbrueckii, Lactobacillus paracasei (Lactobacillus paracasei), Lactobacillus plantarum (Lactobacillus plantarii) and Lactobacillus rhamnosus (Lactobacillus rhamnosus); the microorganism belonging to the genus Lactococcus is Lactococcus lactis; the microorganism belonging to the genus Streptococcus is Streptococcus salivarius (Streptococcus salivarius).
In an embodiment of the present invention, the co-culture ratio of the soybean fermentation broth and the microbial probiotic is 100: 1, and the soybean fermentation broth is non-defatted soybean fermentation broth.
Although the present invention has been described with reference to the above embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention.
Drawings
FIG. 1 illustrates the change in soy fermentation broth gel (FSG) versus wound area. Shows that the soybean fermentation composition (FSG group: soybean fermentation broth gel) has better efficacy on the healing capacity of skin wounds than other experimental samples (Vghile group: blank gel control group, Positive group: steroid-beta pine soft cream BETAMETHASONE oil).
FIG. 2 illustrates the soy fermentation broth gel (FSG) change for wound healing.
FIG. 3, is a schematic representation of a pathological staining assay for wound healing. The mechanism of the soybean fermentation composition (FSG group: soybean fermentation broth gel) for healing skin wounds is to control wound inflammation (similar to steroid) and improve the effects of mature connective tissues and angiogenesis so as to achieve the effect of promoting wound healing.
FIG. 4 illustrates an embodiment of the present invention for wound healing in a human.
FIG. 5 illustrates an embodiment of the present invention for wound healing in a human.
FIG. 6 illustrates an embodiment of the present invention for wound healing in a human.
FIG. 7 illustrates an embodiment of the present invention for wound healing in a human.
FIG. 8 illustrates an embodiment of the present invention for wound healing in a human.
Detailed Description
In order to evaluate whether the soybean fermentation broth contained in the composition of the present invention has the efficacy of promoting the healing of skin wounds, a test evaluation is performed.
Example 1 preparation of Soybean fermentation broth contained in the composition of the present invention
The invention is to grind non-defatted non-genetically modified soybeans into pulp by using a pulping machine, add black sugar and mineral salt into soybean liquid with the ratio of the soybeans to water of 1: 16, heat the mixture and obtain a soybean liquid culture medium without filtering for culturing microbial probiotics. Respectively carrying out liquid culture on the soybean liquid culture medium and microorganisms of Bifidobacterium (Bifidobacterium), Bacillus (Bacillus), Lactobacillus (Lactobacillus), Lactococcus (Lactobacillus) and Streptococcus (Streptococcus) in a Lactobacillus fermentation method (like fermentation or fermentation with different fermentation), adding the soybean liquid culture medium for amplification culture, finally mixing all strains together, adding the soybean liquid culture medium for final culture; wherein the final culture takes a vacuum bag as a plane bag type fermentation-fermentation container, the fermentation-fermentation temperature is controlled at 38 ℃, and the fermentation-fermentation time is more than 168 hours.
The microorganism belonging to the genus Bifidobacterium is Bifidobacterium or Bifidobacterium longum; the microorganism belonging to the genus Bacillus is Bacillus coagulans (Bacillus coagulans); the microorganism of the genus Lactobacillus is selected from the group consisting of Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus casei (Lactobacillus casei), Lactobacillus delbrueckii (Lactobacillus delbrueckii), Lactobacillus paracasei (Lactobacillus paracasei), Lactobacillus plantarum (Lactobacillus plantarii) and Lactobacillus rhamnosus (Lactobacillus rhamnosus); the microorganism belonging to the genus Lactococcus is Lactococcus lactis (Lactococcus lactis); and the microorganism belonging to the genus Streptococcus is Streptococcus salivarius (Streptococcus salivarius).
In the above embodiment, the ratio of co-culturing the soybean liquid and the microbial probiotic group is 100: 1, wherein the bifidobacterium accounts for 0.10-0.30% of the total volume, and the bifidobacterium longum accounts for 0.05-0.15% of the total volume; the bacillus coagulans accounts for 0.10-0.30% of the total volume; the lactobacillus acidophilus accounts for 0.10-0.30% of the total volume, the lactobacillus casei accounts for 0.10-0.30% of the total volume, the lactobacillus delbrueckii accounts for 0.10-0.30% of the total volume, the lactobacillus paracasei accounts for 0.100-0.30% of the total volume, the lactobacillus plantarum accounts for 0.100-0.30% of the total volume, and the lactobacillus rhamnosus accounts for 0.100-0.30% of the total volume; the lactococcus lactis accounts for 0.05-0.15% of the total volume, and the streptococcus salivarius accounts for 0.05-0.15% of the total volume.
In the above embodiment, after the final cultured soybean fermentation broth is heated, sterilized and filtered, the concentration of soluble solids can reach about 5.0Brix, so as to obtain the soybean fermentation broth contained in the composition of the present invention.
Adding appropriate excipient into the fermented soybean fermentation broth, and making into spray, gel, emulsion, cream, ointment, etc. Preferably, the Soybean fermentation compositions for skin wound healing of the present invention are in the form of gels and sprays, namely, Fermented Soybean Filter Gel (FSG) and Fermented Soybean Filter Liquid (FSL).
Example 2 animal experiment module for skin wound healing using the composition of the invention
In order to evaluate whether the soybean fermentation broth contained in the composition of the present invention has the efficacy of promoting the healing of skin wounds, a test evaluation is performed.
This example employed approximately 6-week old male ICR strain mice (purchased from lesco biotechnology, inc.) of approximately 8 individuals per group weighing approximately 30 to 35 grams, and given a 7-day acclimation period, mice were provided free feeding with mouse feed (Chow 5001) and water. The experiments of this example are grouped as follows:
(1) FSG group: namely the experimental group of the invention, the soybean fermentation broth is added with the Gel prepared by the excipient, namely the feedstuff Soybean Filter Gel (FSG).
(2) Vehicle group: water is used to replace the soybean fermentation broth, and the gel prepared by the excipient with the same content in the experimental group is added.
(3) Positive group: a commercially available drug (steroid Betamethsonisomentine, soft extract of Betacathon) was purchased as Positive Control.
The animals are fasted for more than 8 hours a day before the creation, and the drinking water is not limited. Firstly, gasifying isoflurane by using a gas anesthesia instrument, stably putting the experimental animal into a deep anesthesia stage, shaving hair on the back side of an operation area, smearing a part to be cut with excellent iodine, and wiping off residual excellent iodine by using 75% alcohol. Lidocaine (3mg/kg) is injected into the wound part for local anesthesia, the operation is started after 5 minutes, the square wound (1 centimeter multiplied by 1 centimeter) of the whole cortex is wound, only a single wound is opened on the back side of each mouse, the experimental gel administration and the photo shooting record are carried out, and finally, the wound is covered by gauze and a protective cover is worn.
Immediately after the first wound was opened, different experimental samples (FSG group; Vehicle group; Positive group) were given to the wound and the same experimental sample was given continuously with time-based change and wound record (see FIG. 1). The Day of wound initiation is defined as Day 0, and then the wound is subjected to drug replacement and recording every 2 to 3 days; the experimental gel replacement, wound cleaning and recording and other processes are carried out after the experimental animals stably enter a deep anesthesia stage (as shown in figure 2).
Wound area measurement: the area of wound healing was calculated by software (Image J) analysis and the percentage of wound healing was calculated. Percent wound healing ═ wound area on day 0-wound area on day N)/wound area on day 0 x 100%.
Results of wound area change: (see FIG. 1), the wound was found to have a slight area increase due to inflammation in Day 2; differences between groups were exhibited starting at Day4, with the FSG group being significantly lower than the Vehicle group (P ═ 0.006), but the Positive group was not significantly different from the FSG and Vehicle groups, respectively (P > 0.05); significant differences between groups were seen between Day7, FSG group was significantly lower than that of Positive and that of Vehicle (P0.015; P < 0.0001), and Positive group was also significantly lower than that of Vehicle (P0.025); greater interclass differences were exhibited between Day9 wound area groups, with the FSG group being significantly lower than the Positive and Vehicle groups (P0.045; P0.001), and the Positive group also being significantly lower than the Vehicle group (P0.048); day11 showed differences between groups, with the FSG group being significantly lower than the Vehicle group (P ═ 0.006), but the Positive group was not significantly different from the FSG and Vehicle groups (P > 0.05); at the time points of Day14, 17, and 21, there was an inter-group difference between groups, and the Vehicle group was significantly higher than the Positive and FSG groups (P < 0.05), but the Positive and FSG groups were not significantly different (P > 0.05).
The above results show that the soybean fermentation composition (FSG group: soybean fermentation broth gel) of the present invention has better efficacy for skin wound healing than other experimental samples (Vghile group: blank gel control group, Positive group: steroid-beta Person Soft OINTMENT BETAMETHASONE oil).
Example 3 Effect of the compositions of the invention on the histopathology of skin wounds
In the experiment described in example 2 above, 2 animals were sacrificed for each group at Day9 and tissue was taken over the wound area for histopathological sectioning and analysis. (see fig. 3)
Re-epithelialization (H & E stabilizing) assessment (fig. 3, row 1) is an important indicator of wound healing in the epidermal layer of the skin, and results show that the results in the FSG and Positive groups are more favorable than the Vehicle group.
In the evaluation aspect of granulation tissue, a large amount of proliferated microvessels, fibrous connective tissue and inflammatory cells can be seen in the wound healing proliferation stage, so the area size of the granulation tissue has significance for wound healing. This experiment uses MassonTrichrome staining (see row 2 of FIG. 3) to differentiate granulation tissue from wounds, and after observation, the areas of granulation tissue in both FSG and Positive groups were found to be smaller than that in Vehicle, indicating that it may have the effect of slowing the inflammatory response and accelerating wound healing.
Fibrous connective tissue evaluation, more mature connective tissue for wound healing means that entry into the anaphase of granulation group repair has begun, so the results of the above Masson Tricheome staining were further verified by differentiating different types of fibrous connective tissue using Van Gieson staining (fig. 3, row 3), and after observation, it was found that mature granulation tissue in both FSG and Positive groups was more abundant than in Vehicle group.
When the area around the wundbed was observed under a 400-fold visual field with an optical microscope (see FIG. 3, row 4), the number of newly formed blood vessels was found to be significantly greater in both the FSG group and the Positive group than in the Vehicle group.
According to the pathological section analysis, the soybean fermentation composition (FSG group: soybean fermentation broth gel) of the invention has the mechanism of controlling wound inflammation (similar to steroid) and improving the effects of mature connective tissue and angiogenesis for the healing of skin wounds, thereby achieving the effect of promoting the healing of the wounds.
EXAMPLE 4 practical application of the compositions of the present invention to human skin wound healing
Case 1: (see fig. 4) head and neck cancer is treated by radiation, the radiotherapy is suspended due to the discomfort of burning and skin breaking of the skin, the soybean fermentation composition is used on the surface of the skin for 2 times a day, and the skin wound is healed after 5 days.
Case 2: (see fig. 5) head and neck cancer is also treated by radiation, the radiation therapy is suspended due to the discomfort of burning and skin breaking of the skin, the soybean fermentation composition is used on the surface of the skin for 2 times a day, and the skin wound is well healed after 13 days.
Case 3: (see fig. 6) the foot abrasion did not heal after two weeks, and the skin wound healed after 5 days and 13 days after 1 day of application of the soybean fermentation composition of the present invention to the skin surface.
Case 4: (see fig. 7) the diabetic foot wound can not heal for two years, after the soybean fermentation composition is used on the skin surface, 1 time per day, the skin wound heals after 27 days to show a healing trend, only a small hole is left, and the skin wound heals completely after 52 days.
Case 5: (fig. 8) bedsore wounds of long-term bedridden patients are known to be difficult to treat, even most patients are more and more serious, most of the patients can only delay the continuous deterioration even if receiving treatment, and the patients want to heal the bedsore wounds easily, wherein the patients use the soybean fermentation composition on the skin surface 1 time every day and heal the skin wounds to 0.5 square centimeter of surface wounds after 181 days.
Claims (4)
1. A composition for healing skin wound is soybean fermentation broth, which is prepared by co-culturing soybean fermentation broth and more than one microbial probiotic group, wherein the microbial probiotic group is a group consisting of microorganisms of the genera Bifidobacterium (Bifidobacterium), Bacillus (Bacillus), Lactobacillus, and Streptococcus (Streptococcus).
2. The composition of claim 1, wherein the soy fermentation broth is non-defatted soy fermentation broth.
3. The composition of claim 1, wherein the soy fermentation broth is formulated with suitable excipients as a spray, gel, emulsion, cream, ointment, etc.
4. The composition of claim 1, wherein the soy fermentation broth is effective in controlling wound inflammation (steroid-like) and enhancing the neogenesis of mature connective tissue and blood vessels, thereby promoting wound healing.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20120027654A (en) * | 2010-09-13 | 2012-03-22 | (주)아모레퍼시픽 | Cosmetic composition containing extracting fraction of fermented beans as an active ingredient |
| KR20140018666A (en) * | 2012-08-03 | 2014-02-13 | (주)아모레퍼시픽 | Composition for skin external application containing fermented soybean extract |
| CN105395567A (en) * | 2014-09-10 | 2016-03-16 | 宝耕生技股份有限公司 | Composition for relieving side effects of chemotherapeutic drugs |
| KR20180025826A (en) * | 2016-08-31 | 2018-03-09 | 전북대학교산학협력단 | Fish Sauces With Anti-inflammatory Effect Comprising Curcuma Longa L. |
| WO2019143055A1 (en) * | 2018-01-18 | 2019-07-25 | 코스맥스 주식회사 | Composition, comprising extract of fermented beans obtained from fermentation by aspergillus fumigatus strain, for skin care improvement |
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- 2019-09-24 CN CN201910907899.8A patent/CN110755472A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20120027654A (en) * | 2010-09-13 | 2012-03-22 | (주)아모레퍼시픽 | Cosmetic composition containing extracting fraction of fermented beans as an active ingredient |
| WO2012036414A2 (en) * | 2010-09-13 | 2012-03-22 | (주)아모레퍼시픽 | Cosmetic composition comprising a fermented soybean extract fraction as an active ingredient |
| KR20140018666A (en) * | 2012-08-03 | 2014-02-13 | (주)아모레퍼시픽 | Composition for skin external application containing fermented soybean extract |
| CN105395567A (en) * | 2014-09-10 | 2016-03-16 | 宝耕生技股份有限公司 | Composition for relieving side effects of chemotherapeutic drugs |
| KR20180025826A (en) * | 2016-08-31 | 2018-03-09 | 전북대학교산학협력단 | Fish Sauces With Anti-inflammatory Effect Comprising Curcuma Longa L. |
| WO2019143055A1 (en) * | 2018-01-18 | 2019-07-25 | 코스맥스 주식회사 | Composition, comprising extract of fermented beans obtained from fermentation by aspergillus fumigatus strain, for skin care improvement |
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