TWI815349B - Use of mangosteen fruit shell extract in the preparation of a medicament for promoting wound healing in diabetes - Google Patents

Abstract

A use of a composition in preparation of a medicament for promoting wound healing in diabetes is provided, wherein the composition comprises an effective amount of mangosteen fruit shell extract.

Description

Use of mangosteen husk extract for preparing medicine to promote diabetic wound healing

The present invention relates to the use of a composition for preparing a medicament for promoting diabetic wound healing.

The skin is the largest organ of the human body, and there are many types of skin diseases. Skin diseases may be acute (lasting only minutes to hours) or chronic conditions, which may affect an individual for days, months, years or even a lifetime. Skin diseases may It may be a fungal, bacterial, or viral condition, or it may be a non-infectious, immune response, such as an inflammatory reaction with or without allergens, or it may be an idiopathic disease. As a result, symptoms may vary and may range from mild itching, redness and swelling to severe pus and open pain, such as damaging sores, and skin disorders may substantially impact an individual's quality of life.

Diabetes mellitus (DM) is a metabolic disease caused by impaired insulin and/or insulin production leading to hyperglycemia. DM patients are often accompanied by many complications, one of which is impaired self-healing ability.

Diabetic wound healing impairment is related to multiple factors including vascular, neuropathy, immune, and biochemical components, all of which are caused by hyperglycemia. Hyperglycemia stiffens blood vessels, leading to slower circulation, microvascular dysfunction, and reduced tissue oxygenation; hyperglycemia also reduces leukocyte migration into wounds, making the patient more susceptible to infection. Peripheral neuropathy in DM may result in numbness of the area and a reduced ability to feel pain, which may result in the wound not being immediately noticed without appropriate treatment, leading to wound chronicity (Spampinato SF, Caruso GI, De Pasquale R , Sortino MA, Merlo S. The Treatment of Impaired Wound Healing in Diabetes: Looking among Old Drugs. Pharmaceuticals (Basel). 2020; 13(4): 60.).

Of particular note, diabetic foot ulcer (DFU) is a major complication of DM, occurring in 15% of diabetic patients. Risk factors associated with DFU are thought to be neuropathy, vascular disease, and infection. Past studies have also found that diabetic patients have a risk of lower limb amputation that is 15 to 46 times higher than that of non-diabetic patients.

Clinical treatment of DFU should include blood glucose control, reduction of plantar pressure, antibiotics, improvement of peripheral blood circulation, wound dressing, and surgery.

Mangosteen has been studied and applied in the fields of breast cancer prevention and muscle-related diseases. It has also been developed as a nutritional supplement and cosmetic for daily life. It has also been used to treat acute hepatitis, liver fibrosis and prevent cirrhosis.

Matsumoto et al. also studied the purification of α-mangostin, β-mangostin, γ-mangostin, and methyl-β-mangostin from mangosteen husks, and studied the effects of these compounds on the cell cycle. The inhibitory effects at each stage show that the compound has anti-cell proliferation and anti-tumor effects (Bioorg. Med. Chem. 2005, 13, 6064-6069).

The present invention provides the use of a composition for preparing a pharmaceutical composition for treating skin diseases.

Specifically, the present invention provides a composition for preparing diabetic wounds that promote Use of healing medicine, wherein the composition contains an effective dose of mangosteen husk extract. The drug may also be used as a topical treatment, medical device or precision therapy.

Mangosteen husks consist of a softer inner husk and a harder outer husk.

In a preferred embodiment, the mangosteen husk is extracted using a solvent, and the extraction solvent is selected from the group consisting of methanol, ethanol, n-propanol, 2-propanol, n-butanol, acetone, ethyl acetate and water. group.

In another preferred embodiment, the mangosteen husk extract includes mangosteen husk water extract and/or mangosteen husk alcohol extract.

In a preferred embodiment, the mangosteen husk extract is a water extract of mangosteen husk.

In another preferred embodiment, the mangosteen husk extract is a mangosteen husk alcohol extract.

In a preferred embodiment, the mangosteen husk includes the outer husk/inner husk of mangosteen husk and/or the whole husk of mangosteen husk.

In another preferred embodiment, the mangosteen husk is the inner husk of the mangosteen husk.

In a preferred embodiment, the composition of the present invention can be an oral or parenteral preparation. The parenteral preparation can be a topical preparation. The topical preparation can be a cream, cream, ointment, gel, lotion, etc. agent or patch.

In a preferred embodiment, the mangosteen husk extract of the present invention contains α-mangostin and γ-mangostin.

In another preferred embodiment, the mangosteen husk aqueous extract of the present invention contains α-mangostin and γ-mangostin.

In another preferred embodiment, the mangosteen husk alcohol extract of the present invention contains α-mangostin and γ-mangostin.

The "effective dose" as used in the present invention refers to a dose that achieves an effective result when administered to an individual, or a dose that has the required activity in vivo or in vitro. In the case of wound healing, effective clinical results include reducing wound severity, and/or prolonging the individual's lifespan, and/or improving the individual's quality of life compared to no treatment. The precise amount of compound administered to an individual will depend on the type and severity of the disease or condition, as well as individual characteristics such as the individual's general health, age, gender, weight, and tolerance to the drug. It also depends on the severity and type of the wound. Those skilled in the art will be able to determine appropriate dosages based on these and other factors.

In one embodiment, the effective dose of the mangosteen husk extract of the present invention is 0.5% (w/w) to 20% (w/w); in a preferred embodiment, the mangosteen husk extract of the present invention The effective dose is 1% (w/w) to 15% (w/w); in the best implementation, the effective dose of the mangosteen shell extract is 1.25% (w/w) to 10% (w/ w).

The pharmaceutical composition of the present invention can be formulated into various oral or parenteral preparations. Oral preparations can be formulated into solid preparations, such as powders, granules, tablets, capsules, etc., or into liquid preparations, such as suspensions, emulsions, syrups, etc. Parenteral preparations can be formulated into topical preparations, such as creams, ointments, gels, lotions, patches, etc., or inhalants, aerosols, suppositories, etc.

The pharmaceutical composition of the present invention may contain pharmaceutically acceptable excipients, especially a predetermined solvent or oil, and may further contain a dispersant if necessary.

Examples of solvents used in the present invention include, but are not limited to, water, ethanol, isopropyl alcohol, 1,3-butanediol, propylene glycol, glycerol, etc.

Examples of oils useful in the present invention are selected from the group consisting of corn oil, sesame oil, linseed oil, cottonseed oil, soybean oil, peanut oil, mono-glycerides, di-glycerides, tri-glycerides, mineral oil, and deep-sea fish mackerel. Squalene, jojoba oil, olive oil, evening primrose oil, borage oil, grape seed oil, coconut oil, sunflower oil, shea butter and any combination thereof groups, but not limited to this.

Solvents and oils can be used alone or in any combination.

Examples of useful dispersants may include lecithin, organic monoglycerides, sorbitan fatty acid esters, polyoxyethylene fatty acid esters, sorbitan stearate, etc., but are not limited thereto. These raw materials can also be used alone or in any combination.

If desired, the composition may further comprise additional ingredients, such as antimicrobial agents or preservatives.

At the same time, conventional active ingredients can be used simultaneously with the composition, as long as they do not have any adverse effect on the pharmaceutical activity of the composition of the present invention. For example, moisturizers such as ceramide, commonly known as atopic dermatitis agents, or liquid ingredients such as steroids such as hydrocortin, vitamin A derivatives such as vitamin A palmitate and/or Tocopherols and the like can be used with the composition.

When the pharmaceutical composition is used as an external preparation, an appropriate skin external preparation can be used as the basic raw material. Aqueous solutions, non-aqueous solvents, suspensions, emulsions or freeze-dried preparations can be used and sterilized according to common methods.

In the actual administration or administration of the composition of the present invention, the dosage may be determined according to various factors such as the route of administration, age, gender, and weight of the patient, as well as the severity of the disease and the dosage form as the active ingredient.

Where the composition of the invention may be a food or cosmetic composition, the composition may be prepared by the appropriate addition of at least one food nourishing or cosmetically acceptable carrier.

The food composition can be used in or added to, for example, health foods. As used herein, the term "healthy food" means a food containing the composition of the present invention that has enhanced functions compared to ordinary food. Healthy foods can be prepared by adding the composition to general foods, or by encapsulation, powdering or suspension.

The cosmetic composition may be added by itself or together with other cosmetic ingredients, or may be used appropriately according to other conventional methods. Cosmetics include, but are not limited to, aftershaves, lotions, creams, facial masks and makeup.

Cosmetic compositions can be formulated into various composition forms, such as gels, creams, ointments, and the like. Compositions in the form of gels, creams and ointments may be suitably prepared according to the form of the composition using known methods by adding conventional softeners, emulsifiers and thickeners or other raw materials known in the art.

Compositions in gel form can be prepared, for example, by adding softeners such as trimethyl alcohol propane, polyethylene glycol and glycerol, solvents such as propylene glycol, ethanol and isocetyl alcohol, and pure water.

Compositions in cream form can be prepared, for example, by adding fatty alcohols, such as stearyl alcohol, myristyl alcohol, behenyl alcohol, arachidyl alcohol, isostearyl alcohol and isocetyl alcohol; emulsifiers, such as lipids , such as lecithin, phosphatidyl choline, phosphatidyl ethanolamine, phospholipid serine, phosphatidylinositol and its derivatives, glyceryl stearate, sorbitol palmitate, sorbitol stearate, etc.; natural fats and oils, such as avocado oil, almond oil, babassu oil, borage oil, camellia oil, etc.; lipid compositions, such as ceramide, cholesterol, fatty acids, phytosphingosine, lecithin, etc.; Solvents, such as propylene glycol, etc.; and pure water.

Compositions in the form of ointments can be prepared, for example, by adding softeners, emulsifiers and waxes, such as microcrystalline wax, paraffin, ceresin, beeswax, spermaceti, petroleum jelly and the like.

In another aspect, the present invention provides a method of using the composition to prepare a medicament for treating or alleviating delayed wound healing in diabetes. As used herein, the term "treatment or alleviation" means halting or delaying the course or symptoms of a disease when administered to a patient.

Figure 1 shows the wound healing efficacy of each test article of the present invention compared to the disease control group and the normal control group. 1A : 1.25% TA1; 1B : 2.5% TA1; 1C : 5% TA1; 1D : 2.5% TA2; 1E : 5% TA2; 1F : 2.5% TA3; 1G : 5% TA3.

Figure 2 shows the wound healing efficacy of 10% TA1 in the present invention compared with the disease control group and the normal control group. .

The following embodiments are non-limiting and merely represent aspects and features of the invention.

Experimental example

Material

Preparation of pharmaceutical compositions

Take the mangosteen husk, dry the husk to 50%~95%, extract it with a solvent (such as water or 10%~95% alcohol), and concentrate to obtain the mangosteen husk extract.

Separate the outer husk and inner husk of the mangosteen husk, dry the outer husk and inner husk of the mangosteen husk respectively to 50%~95%, and use a solvent (such as water or 10%~95% alcohol) Carry out extraction and concentration to obtain the outer husk extract of mangosteen husk and the inner husk extract of mangosteen husk.

The alcohol and water extracts of mangosteen whole shell (test product 1; TA1), mangosteen outer shell (test product 2; TA2), and mangosteen inner shell (test product 3; TA3) were prepared into different concentrations. of cream or ointment.

experimental animals

The present invention uses adult (7-8 weeks old) male Crl:CD ^® (SD) rats with an average body weight of 200-250 g. Ear-notch and cage tags are used for animal identification. Animals were housed individually in polycarbonate cages under the following conditions: temperature maintained at 22 ± 3 °C; humidity maintained at 50 ± 20%; 12 h/12 h light/dark cycle. Food and water were provided ad libitum throughout the study.

method

Streptozotocin (STZ)-induced diabetes model

A total of 84 rats were used in this invention, of which 12 rats were used as normal controls, and the remaining 72 rats were injected intraperitoneally (IP) with a single dose (50 mg/kg) of streptozotocin (STZ; Sigma Aldrich S0130 ) to induce type I diabetes. STZ is freshly prepared in 0.1M citrate buffer (pH=4~4.5) to a concentration of 5mg/mL.

At 2, 4, 6, and 7 weeks after STZ administration, the fasting blood glucose level of each STZ-induced rat was measured using a blood glucose meter. Diagnostic criteria for diabetes in this model include: fasting blood glucose concentration >250 mg/dL, weight loss, polyuria, and polydipsia.

One week after the diagnosis of diabetes, all diabetic rats were weighed, fasting blood glucose concentrations were measured, and any rats showing blood glucose concentrations <250 mg/dL or weight gain were removed from the study.

Fifty-four rats were divided into 9 groups (6 rats in each group) for use in the study of the present invention. The groups, treatment methods, test articles and number of animals in each group are as shown in the table below:

To further evaluate the wound healing efficacy of higher dose test items, the present invention also administered 10% of TA1, TA2 and TA3 to rats. A total of 30 male rats (6 rats in each group) were used for this test. and carry out the same experimental process. The group number, treatment, test article and number of animals in each group are as shown in the table below:

Creation of wound model

STZ-induced diabetic rats or healthy rats were anesthetized, the backs of all rats were shaved, and full-thickness excision wounds were made at the panniculus carnosus muscle layer. (20mm×20mm). The wound is not sutured or covered, but secondary healing is allowed. Wounds were inflicted in the same manner on all study rats.

Topical administration

Each test product (whole husk extract, inner husk extract, and outer husk extract) or vehicle was topically applied to the entire wound once a day in a film-like form with a thickness of approximately 0.25 mm (total 0.1g/wound). The wound was gently cleaned with normal saline before each application; a total of 21 administrations were performed per rat.

Animal observations will be made for all surviving study rats at least once daily, and body weights will be recorded on Day 0 (pre-dose), at least weekly, and on the day of necropsy.

Wound measurement

During the period of topical administration, gross observations of each wound were performed daily. Wound area was measured in two different ways: (i) by tracing on transparent plastic sheets every 3 days, and (ii) by recording digital images with a digital camera every 3 days.

Wound area was calculated using Image J software. Initial post-trauma (Day 0) wound area was used to calculate percent wound closure for each wound at each dosing date. The calculation formula for the percentage of wound area covered by new granulation tissue is {[(Area _i (A _i )-Area _n (A _n )]/Area _i (A _i )}×100, where A _i is the initial wound area, A _n is the wound area on the nth day.

analyze

This invention uses SigmaPlotTM Statistical Software for WindowsTM, Release 12.0 software (Jandel Scientific Inc., USA) for statistical analysis, and all statistical tests use a significance standard of 0.05.

result

diabetes animal model

All rats used in the present invention showed characteristic signs of hyperglycemia on the second day after streptozotocin (STZ) induction. 48 hours after STZ induction, the blood glucose concentrations of rats in all groups increased significantly and remained elevated throughout the experiment (see Table 1). Until the end of the animal experiment, all groups except group 1 All rats were in a diabetic state.

Table 1

wound healing

The following results were obtained during wound healing studies in control and diabetic experimental rats (Table 2).

Table 2

Wound areas measured at different times after wound creation, namely days 3, 6, 9, 12, 15, 18 and 21, showed that topical administration of the test articles (TA1, TA2 and TA3) had an effect on diabetic wounds. The healing of the mouth has a positive effect. It was observed in the present invention that the wound closure rate of the disease control rats (Group 2) was reduced compared with the normal control rats (Group 1), which shows that the chronic wound healing model used in the present invention has been validated, and The effect can be seen more obvious from the 3rd day onwards.

As shown in Figure 1, compared with the disease control rats (Group 2), the experimental rats that were topically administered TA1 from the 15th day (Group 3: 1A, Group 4: 1B and Group 5: 1C )'s wound contraction ability shows obvious wound healing ability. While on day 21, the maximum wound healing rate (percentage) was observed in 5% TA1 treated animals (Group 5) compared to disease control rats (Group 2): Wound healing rates were 63.18± 10.65 (Group 2) and 88.35±2.68 ( P <0.05) (Group 5).

Compared with disease control rats (Group 2), the wound shrinkage ability of experimental rats topically administered TA2 (Group 6: 1D and Group 7: 1E) showed significant wound healing from day 15 onwards.

Compared with disease control rats (Group 2), the wound shrinkage ability of experimental rats topically administered TA3 (Group 8: 1F and Group 9: 1G) showed significant wound healing from day 15 onwards.

In summary, the research of the present invention found that the extracts of mangosteen whole husk (TA1), mangosteen outer husk (TA2) and mangosteen inner husk (TA3) all have the potential to promote wound healing because they With better epidermal and dermal regeneration, as well as granulation tissue production and epidermal migration.

The results pointed out that TA1, TA2 and TA3 significantly improved the symptoms of slow wound healing in diabetic rats. Among them, 5% mangosteen whole husk, 2.5% and 5% mangosteen inner husk have the best wound healing effect and reduce the concentration of pro-inflammatory cytokines, so they are considered the best formula. Compare Compared with the three test products with the same concentration (5%), the order of wound healing ability is that the inner shell of mangosteen fruit is the best, followed by the whole mangosteen shell, and finally the outer shell of mangosteen fruit.

Regarding the wound healing effect results of 10% TA1, TA2 and TA3, compared with the disease control rats, 10% TA1 had a significant difference in wound healing from the 12th day (Figure 2), among which the wound healing effect of 10% TA1 was superior. At 5% TA1. TA2 and TA3 also showed the same trend. In order of wound healing ability, the inner shell of mangosteen fruit was the best, followed by the whole mangosteen shell, and finally the outer shell of mangosteen fruit.

While the invention has been described and exemplified in sufficient detail to enable those of ordinary skill in the art to make and practice it, it will be apparent that various substitutions, modifications and improvements can be made without departing from the spirit and scope of the invention. of.

It will be readily understood by those of ordinary skill in the art that the present invention is well adapted to carry out the stated objects and to obtain the objects and advantages mentioned and those inherent therein. The cells, animals, and processes and methods for producing them represent only the best embodiments, are illustrative, and are not intended to limit the scope of the invention. Modifications and other uses can be conceived by those of ordinary skill in the art, and these modifications are included in the spirit of the invention and are limited by the scope of the patent application.

Claims (8)

A composition is used to prepare a medicament for promoting diabetic wound healing, wherein the composition includes an effective dose of mangosteen husk extract, wherein the mangosteen husk extract is a mangosteen husk extract. The use as described in claim 1, wherein the mangosteen husk extract contains mangosteen endohusk water extract and/or mangosteen endohusk alcohol extract. The use as described in claim 1, wherein the mangosteen husk extract contains α-mangostin (α-mangostin) and γ-mangostin (γ-mangostin). The use as claimed in claim 1, wherein the composition is a parenteral preparation. The use as claimed in claim 4, wherein the parenteral preparation is an external preparation. The use as claimed in claim 1, wherein the effective dose is 0.5% w/w to 20% w/w. The use as claimed in claim 1, wherein the effective dose is 1% w/w to 15% w/w. The use as claimed in claim 1, wherein the effective dose is 1.25% w/w to 10% w/w.

Images