CN116763832A

CN116763832A - Application of mangosteen shell extract in preparation of medicine for promoting diabetic wound healing

Info

Publication number: CN116763832A
Application number: CN202210233876.5A
Authority: CN
Inventors: 陈谷政; 陈衍儒; 陈诗茵; 庄苡品
Original assignee: Xantho Biotechnology Co ltd
Current assignee: Xantho Biotechnology Co ltd
Priority date: 2022-03-10
Filing date: 2022-03-10
Publication date: 2023-09-19

Abstract

The invention provides a use of a composition for preparing a medicament for promoting diabetic wound healing, wherein the composition comprises an effective dose of mangosteen shell extract.

Description

Application of mangosteen shell extract in preparation of medicine for promoting diabetic wound healing

Technical Field

The invention relates to application of a composition in preparing a medicament for promoting diabetic wound healing.

Background

Skin is the largest organ of the human body, skin diseases are also numerous, skin diseases may be acute (lasting only minutes to hours) or chronic conditions, which may affect individuals for days, months, years or even life, skin diseases may be fungal, bacterial, or viral conditions, or may be non-infectious, immune reactions, such as inflammatory reactions with or without allergens, or may be idiopathic. Thus, symptoms can be varied and can range from mild itching, redness and swelling to severe purulent and open pain, such as a nociceptive ulcer, skin conditions can substantially affect the quality of life of an individual.

Diabetes Mellitus (DM) is a metabolic disease, which is caused by impaired insulin and/or insulin producing functions leading to hyperglycemia. DM patients are often associated with a number of complications, one of which is impaired self-repair.

Diabetic wound healing disorders are associated with a variety of factors, such as blood vessels, neuropathy, immune and biochemical components, and are caused by hyperglycemia. Hyperglycemia causes vascular sclerosis, which in turn leads to slowing of circulation and microvascular dysfunction, and to reduced tissue oxygenation; hyperglycemia also reduces migration of leukocytes to the wound, making the patient more susceptible to infection. Whereas peripheral neuropathy in DM may lead to numbness in the area and reduced ability to feel pain, which may lead to the wound not being immediately noticed and appropriate treatment being taken, resulting in 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 Ulcers (DFUs) are a major complication of DM, which occurs in 15% of diabetics. Risk factors associated with DFU are considered to be neuropathies, vascular diseases and infections. Past studies have also found that diabetic patients have 15 to 46 times the risk of amputation of their lower limbs than non-diabetic patients.

Clinical treatments for DFU should include controlling blood glucose, reducing plantar pressure, antibiotics, improving peripheral blood circulation, wound dressing and surgery.

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

Matsumoto et al also studied the purification of alpha-mangostin, beta-mangostin, gamma-mangostin, and methyl-beta-mangostin from mangosteen hulls, and studied the inhibition of the compounds on various stages of the cell cycle, indicating that the compounds have anti-cell proliferation and anti-tumor effects (bioorg. Med. Chem.2005,13, 6064-6069).

Disclosure of Invention

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

In particular, the invention provides a use of a composition for preparing a medicament for promoting diabetic wound healing, wherein the composition comprises an effective dose of mangosteen shell extract. The medicament may also be used for topical treatment, medical devices or precision treatment.

The mangosteen rind comprises a softer inner rind and a harder outer rind.

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

In another preferred embodiment, the mangosteen pod extract comprises a mangosteen pod water extract and/or a mangosteen pod alcohol extract.

In a preferred embodiment, the mangosteen pod extract is a mangosteen pod aqueous extract.

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

In a preferred embodiment, the mangosteen rind comprises the outer/inner rind of the mangosteen rind and/or the whole rind of the mangosteen rind.

In another preferred embodiment, the mangosteen rind is an outer rind of a mangosteen rind.

In a preferred embodiment, the composition of the present invention may be an oral or parenteral formulation, which may be an external formulation, which may be a cream, an ointment, a gel, a lotion or a patch.

In a preferred embodiment, the mangosteen rind extract of the invention comprises alpha-mangostin (α -mangostin) and gamma-mangostin (γ -mangostin).

In another preferred embodiment, the mangosteen rind aqueous extract of the invention comprises alpha-mangostin (alpha-mangostin) and gamma-mangostin (gamma-mangostin).

In yet another preferred embodiment, the mangosteen hull alcohol extract of the present invention comprises alpha-mangostin (alpha-mangostin) and gamma-mangostin (gamma-mangostin).

An "effective dose" as referred to herein is a dose that achieves an effective result when administered to an individual, or a dose that possesses a desired activity in vivo or in vitro. In the case of wound healing, effective clinical outcomes include slowing the severity of the wound, and/or extending the life of the individual, and/or improving the quality of life of the individual, as compared to untreated. The precise amount of compound administered to an individual will depend on the type and severity of the disease or condition and the individual characteristics, such as the general health, age, sex, weight and tolerance to drugs of the individual, and also on the wound severity and type. One skilled in the art will be able to determine the appropriate dosage based on these and other factors.

In one embodiment, the effective dose of mangosteen rind extract of the invention is from 0.5% (w/w) to 20% (w/w); in a preferred embodiment, the effective dose of mangosteen rind extract of the present invention is 1% (w/w) to 15% (w/w); in a preferred embodiment, the mangosteen pod extract is effective at a dosage of 1.25% (w/w) to 10% (w/w).

The pharmaceutical compositions of the present invention may be formulated in a variety of oral or parenteral formulations. Oral formulations may be formulated as solid formulations, such as powders, granules, lozenges, capsules, etc., or as liquid formulations, such as suspensions, emulsions, syrups, etc. Parenteral formulations may be formulated as external preparations such as creams, ointments, gels, lotions, patches and the like, or as inhalants, aerosols, suppositories and the like.

The pharmaceutical composition of the present invention may comprise pharmaceutically acceptable excipients, and in particular may further comprise a predetermined solvent or oil, if necessary, and may further comprise a dispersing agent.

Examples of solvents used in the present invention include, but are not limited to, water, ethanol, isopropanol, 1, 3-butanediol, propylene glycol, glycerin, and the like.

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, mackerel Squalene (squarene), jojoba Oil (jojoba Oil), olive Oil, evening primrose Oil, borage Oil (Borage Oil), grape seed Oil, coconut Oil, sunflower seed Oil, shea butter, and any combination thereof, but are not limited thereto.

The solvents and oils may be used alone or in any combination thereof.

Examples of useful dispersants may include, but are not limited to, lecithin, organic monoglycerides, fatty acid esters of resveratrol, polyoxyethylene fatty acid esters, sorbitan stearate, and the like. These materials may also be used alone or in any combination thereof.

If desired, the composition may further comprise additional materials, such as an antimicrobial agent or preservative.

Meanwhile, the conventional active ingredient may be used simultaneously with the composition as long as it has no adverse effect on the pharmaceutical activity of the composition of the present invention. For example, lotions such as ceramides (ceramides) are commonly used as conventional atopic dermatitis agents, or liquid ingredients, steroids such as hydroepinephrine, vitamin a derivatives such as vitamin a palmitate and/or tocopherol, and the like may be used with the compositions.

When the pharmaceutical composition is used as an external preparation, an appropriate external preparation for skin can be used as a base material, and an aqueous solution, a nonaqueous solvent, a suspension, an emulsion, a lyophilized preparation or the like can be used and sterilized by a conventional method.

In the compositions of the present invention which are actually administered or administered, the dosage may be determined according to various factors such as the route of administration, age, sex, and weight of the patient, and severity of the disease and the type of medicament as an active ingredient.

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

The food composition may be used or added to, for example, a health food. As used herein, the term "health food" means a food containing the composition of the present invention having an enhanced function compared to general foods. The health food may be prepared by adding the composition to general foods, or by encapsulation, powdering, or suspension liquefaction.

The cosmetic composition may be added as it is or together with other cosmetic ingredients, or may be suitably used according to other conventional methods. The cosmetic includes, but is not limited to, after-shave lotion (after shaves), astringent, cream, facial mask, and make-up.

The cosmetic compositions may be formulated in various composition forms, such as gels, creams, ointments, and the like. Compositions in the form of gels, creams and ointments may be suitably prepared by adding conventional softeners, emulsifiers and thickeners or other materials known in the art, depending on the form of the composition, using known methods.

The gel-form composition can be prepared, for example, by adding a softener such as trimethylol propane, polyethylene glycol, and glycerin, a solvent such as propylene glycol, ethanol, and isocetyl alcohol, and pure water.

The compositions in the form of creams can be prepared, for example, by adding fatty alcohols, such as stearyl alcohol, myristyl alcohol, behenyl alcohol, arachidyl alcohol, isostearyl alcohol and isocetyl alcohol; emulsifying agents, for example, lipids, such as lecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol and derivatives thereof, glyceryl stearate, sorbitol palmitate, sorbitol stearate, and the like; natural fats and oils such as avocado oil, almond oil, babassu oil (babassu oil), borage oil, camellia oil, and the like; lipid compositions such as ceramides, cholesterol, fatty acids, phytosphingosine, lecithins, and the like; solvents such as propylene glycol and the like; and (3) pure water.

The compositions in the form of ointments may be prepared, for example, by adding softeners, emulsifiers and waxes, such as microcrystalline wax, paraffin wax, ceresin (ceresin), beeswax, spermaceti, petrolatum and the like.

In another aspect, the invention provides a method of using the composition to prepare a medicament for treating or alleviating delayed healing of a diabetic wound. As used herein, the term "treating or alleviating" means stopping or delaying the course or symptoms of a disease when the patient is taking the agent.

Drawings

Fig. 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.

Fig. 2 is a graph showing the wound healing efficacy of 10% of the TA1 of the present invention compared to the disease control and normal control.

Detailed Description

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

Experimental example

Material

Preparation of pharmaceutical compositions

Drying mangosteen shells to 50% -95%, extracting with solvent (such as water or 90% + -10% alcohol), and concentrating to obtain mangosteen shell extract.

Separating outer and inner shells of mangosteen, drying the outer and inner shells to 50% -95%, extracting with solvent (such as water or 90% + -10% alcohol), and concentrating to obtain outer and inner shells extract.

Respectively preparing alcohol and water extracts of whole fruit shell (test 1; TA1), outer fruit shell (test 2; TA2) and inner fruit shell (test 3; TA3) into cream or ointment with different concentrations.

Experimental animal

The invention uses adult (7-8 week old) male Crl with an average body weight of 200-250 g:(SD) rats. Ear-notch (ear-notch) and cage tags are used for animal identification. Animals were housed individually in polycarbonate cages under the following conditions: the temperature is kept at 22+/-3 ℃; the humidity is kept at 50+/-20%; light/dark cycle of 12 hours/12 hours. Food and water were provided ad libitum throughout the study.

Method

Streptozotocin (STZ) -induced diabetes model

The present invention used a total of 84 rats, of which 12 were used as normal control groups and the remaining 72 were induced by intraperitoneal Injection (IP) of single dose (50 mg/kg) of streptozotocin (STZ; sigma Aldrich S0130) to induce type I diabetes. STZ was freshly prepared in 0.1M citrate buffer (ph=4-4.5) to a concentration of 5mg/mL.

At weeks 2, 4, 6 and 7 after STZ dosing, fasting blood glucose levels were measured for each STZ-induced rat using a glucometer. Diagnostic criteria for diabetes in this model include: fasting blood glucose concentrations >250mg/dL, weight loss, polyuria and polydipsia.

After one week of diagnosis of diabetes, all diabetic rats were weighed, fasting blood glucose concentrations were measured, and any rats exhibiting a blood glucose concentration <250mg/dL or weight gain were removed from the study.

54 rats were divided into 9 groups (6 rats per group) for the study of the present invention. The group, treatment regimen, test article and animal numbers for each group are shown in the following table:

further evaluation of wound healing efficacy of higher dose test articles, 10% of TA1, TA2 and TA3 were also administered to rats, and a total of 30 male rats (6 rats per group) were used for this test and the same experimental procedure was performed. Group numbers, treatments, test articles and animal numbers for each group are shown in the following table:

group of	Treatment mode	Test article	Number of animals
				10	Wound + vehicle	Medium agent (normal control group)	6 pieces of
11	STZ+wound+vehicle	Vehicle (disease control group)	6 pieces of
				12	Stz+wound+10% TA1	TA1	6 pieces of
13	Stz+wound+10% TA2	TA2	6 pieces of
				14	Stz+wound+10% TA3	TA3	6 pieces of

Wound model creation

STZ-induced diabetic rats or healthy rats were anesthetized, the backs of all rats were shaved, and full-thickness excision wounds (20 mm x 20 mm) were made in the myofibrillar (panniculus carnosus) muscle layer. The wound is not sutured or covered, but is allowed to heal in the second phase. All study rats were wound-inflicted in the same manner.

Topical administration

Each test article (whole, inner and outer shell extracts) or vehicle was applied topically to the entire wound once per day in the form of a film with a thickness of about 0.25mm (total 0.1 g/wound). Lightly cleaning the wound with normal saline before each coating; a total of 21 doses were administered to each rat.

Animal observations were made at least once daily for all surviving study rats, and body weight was recorded on day 0 (pre-dose), at least weekly, and on necropsy.

Wound measurement

During topical administration, each wound was generally observed daily. Wound area is measured in two different ways: (i) Tracking on transparent plastic film once every 3 days, and (ii) recording digital images using a digital camera once every 3 days.

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

Analysis

The present invention uses SigmaPlotTM Statistical Software for WindowsTM, release 12.0 software (Jandel Scientific inc., USA) for statistical analysis, all statistical tests using a 0.05 significance standard.

Results

Animal model for diabetes

All rats used in the present invention exhibited the characteristic signs of hyperglycemia the following day after induction with Streptozotocin (STZ). After 48 hours of STZ induction, blood glucose concentrations were significantly elevated in all groups of rats and maintained in an elevated state throughout the experiment (see table 1) until the termination of the animal experiment, all groups of rats were in a diabetic state except group 1.

TABLE 1

* : p <0.05 (compared to group 2)

Wound healing

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

TABLE 2

The wound areas measured at various times after wound creation, i.e. days 3,6, 9, 12, 15, 18 and 21, showed that topical administration of the test articles (TA 1, TA2 and TA 3) had a positive effect on the healing of diabetic wounds. A reduction in wound closure rate was observed in the present invention in the disease control rats (group 2) compared to the normal control rats (group 1), indicating that the chronic wound healing model used in the present invention was validated and that the effect was more pronounced from day 3.

As shown in fig. 1, the wound-contracting ability of the experimental rats (group 3: 1A, group 4: 1B, and group 5: 1C) topically administered TA1 showed remarkable wound-healing ability from day 15, compared to the disease control rats (group 2). Whereas at day 21, the maximum wound healing rate (percent) was observed in animals treated with 5% ta1 (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), respectively.

The ability of the experimental rats (group 6: 1D and group 7: 1E) to contract wounds with TA2 administered topically showed significant wound healing from day 15 compared to the disease control rats (group 2).

The ability of the experimental rats (group 8: 1F and group 9: 1G) to contract wounds, which were topically given TA3, showed significant wound healing from day 15, compared to the disease control rats (group 2).

In summary, the studies of the present invention have found that extracts of whole mangosteen (TA 1), outer mangosteen (TA 2) and inner mangosteen (TA 3) have the potential to promote wound healing, as they have better epidermis and dermis regeneration, as well as granulation tissue formation and epidermis migration.

The results indicate that TA1, TA2 and TA3 significantly improved the symptoms of slow wound healing in diabetic rats. Among them, 5% of whole mangosteen shells, 2.5% and 5% of inner mangosteen shells have the best wound healing effect, and the concentration of pro-inflammatory cytokines is reduced, which is considered as the optimal formulation. Comparing the three test products with the same concentration (5%), the sequence of wound healing capacity is that the inner shells of the mangosteen are the best, the whole shells of the mangosteen are the next time, and finally the outer shells of the mangosteen.

Results for wound healing effects of 10% TA1, TA2 and TA3 there was a significant difference in wound healing of 10% TA1 from day 12 compared to disease control rats (fig. 2), with the wound healing effect of 10% TA1 being better than 5% TA1.TA2 and TA3 also show the same trend, and the sequence of wound healing capacity is the best of the inner shells of the mangosteen, the second time of the whole shells of the mangosteen, and finally the outer shells of the mangosteen.

While the invention has been described and illustrated in sufficient detail to enable those skilled in the art to make and practice the invention, various alternatives, modifications and improvements should be apparent without departing from the spirit and scope of the invention.

Those skilled in the art will readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The cells, animals, and processes and methods of producing them are merely representative of the best modes for carrying out, are illustrative, and are not intended to limit the scope of the invention. Modifications and other uses thereof will occur to those skilled in the art which are encompassed within the spirit of the invention and are defined by the scope of the claims.

Claims

1. Use of a composition for the manufacture of a medicament for promoting diabetic wound healing, wherein the composition comprises an effective dose of mangosteen shell extract.

2. The use according to claim 1, wherein the mangosteen pod extract comprises a mangosteen pod water extract and/or a mangosteen pod alcohol extract.

3. Use according to claim 1, wherein the mangosteen rind comprises the outer rind of the mangosteen rind and/or the inner rind of the mangosteen rind.

4. The use of claim 1, wherein the mangosteen rind is an inner rind of a mangosteen rind.

5. The use of claim 1, wherein the mangosteen pod extract comprises alpha-mangostin alpha-and gamma-mangostin gamma-.

6. The use of claim 1, wherein the composition is a parenteral formulation.

7. The use according to claim 6, wherein the parenteral formulation is a topical formulation.

8. The use of claim 1, wherein the effective dose is 0.5% w/w to 20% w/w.

9. The use of claim 1, wherein the effective dose is 1% w/w to 15% w/w.

10. The use of claim 1, wherein the effective dose is 1.25% w/w to 10% w/w.

11. The use of claim 1, wherein the wound comprises a diabetic foot ulcer.