WO2018026264A2

WO2018026264A2 - Medicament for wound treatment

Info

Publication number: WO2018026264A2
Application number: PCT/MY2017/050046
Authority: WO
Inventors: Isa BIN NAINA MOHAMED; Ahmad Nazrun BIN SHUID; Nurul Yuziana BINTI MOHD YUSOF; Shiplu Roy CHOWDHURY; Shihab Uddin AHMAD
Original assignee: Universiti Kebangsaan Malaysia
Priority date: 2016-08-04
Filing date: 2017-08-03
Publication date: 2018-02-08
Also published as: WO2018026264A3; MY187799A

Abstract

The present invention discloses the use of Labisia or Marantodes plant extract for the manufacture of a topical medicament for the treatment of a variety of wounds such as incision, abrasion, avulsion, laceration and puncture.

Description

MEDICAMENT FOR WOUND TREATMENT

FIELD OF INVENTION

The present invention relates to a medicament for wound treatment. More particularly, the present invention relates to a topical medicament for wound treatment.

BACKGROUND OF THE INVENTION

Skin is the largest organ of the human body. It is soft to allow movement, but tough enough to resist breaking or tearing. It acts as the body's shield, blocking out germs and bacteria. When the skin is breached either due to accidents or surgical procedures, a wound is created. Wounds are physical injuries that results in an opening and break of the skin. Open wounds such as incision, abrasion, avulsion, laceration or puncture wounds are the most common wounds affecting humans. Common complains or complications following wounds are pain, swelling, inflammation or redness, bleeding, infection and loss of function. Although our body begins to repair an injury immediately, the healing process of wounds will continue for days, weeks, months or even years, depending on the type and size of the injury. Numerous topical medicaments have been developed in connection with the treatment of skin injuries or wounds in order to aid the body's natural healing functions and expedite the skin's healing process. An example is disclosed in United States Patent Publication No. US 2004/0001878 A1. The prior art discloses the treatment of wounds by covering and contacting the wound with a dressing that has been infused with a suspension of a starch hydrolysate containing ascorbic acid, collagen and alpha-tocopherol acetate. However, ascorbic acid may irritate sensitive skin. Therefore, it is not uncommon to see side effects such as inflammation, contact dermatitis and heightened skin sensitivity from the use of a dressing or ointment containing ascorbic acid. The body's natural wound healing itself elicits mechanical stress that activates itching. Coupled with inflammation induced by the dressing or ointment containing ascorbic acid, a patient may be urged to scratch the wound. Scratching a wound that is trying to heal could interfere with the body's own healing mechanism as it can cause damage to the new tissue that has grown to replace and repair the damaged tissue. If this happens, it can slow the healing process, which leaves your body susceptible to wound infection longer and can result in excessive scarring. Additionally, it can cause potentially harmful bacteria on the hands to transfer to the wound, increasing the risk of developing an infection on the wound. The most common wound healing agents currently in use today are alcohol, and iodine including its derivatives povidone and flavine. These are antiseptic agents and will prevent wounds from being infected. It has no inherent wound healing properties. Antiseptics do not promote wound healing.

For the reasons described above, there is a need for a topical medicament for wound treatment that addresses the above mentioned limitation of the existing topical medicaments.

SUMMARY OF INVENTION

The present invention relates a medicament for wound treatment. In a first aspect of the invention, the use of Labisia pumila or Marantodes pumilum plant extract in the manufacture of a medicament for wound treatment is disclosed. Preferably, the Labisia plant extract is in an amount of 0.5 to 3.0 percentage by weight of the medicament.

In a second aspect of the invention, a wound bandage comprising an adhesive member adapted for application to a person's skin and a pad on one side of the adhesive member is disclosed. The pad is impregnated with powdered Labisia pumila or Marantodes pumilum plant extract, wherein the powdered Labisia plant extract is suitably in an amount of 0.5 to 5.0 percentage by weight of the pad. In a third aspect of the invention, a gauze for wound treatment is disclosed.

The gauze comprises a dry textile material impregnated with powdered Labisia pumila or Marantodes pumilum plant extract. Preferably, the powdered Labisia plant extract is in an amount of 0.5 to 5.0 percentage by weight of the textile material. BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate embodiments of the invention and, together with the description, serve to explain the principles of the invention. FIG. 1 illustrates a flowchart of a method for preparing Labisia extract ointment according to an embodiment of the present invention.

FIG. 2 illustrates photographic representations showing wound contraction area on Labisia extract ointment treated rats on post-wounding day 9.

FIG. 3 illustrates a bar chart representing complete healing day for all experimental rat groups. FIG. 4 illustrates a graph representing the percentage of wound contraction for all experimental rat groups.

DESCRIPTION OF THE PREFERRED EMBODIMENT

A preferred embodiment of the present invention will be described herein below with reference to the accompanying drawings. In the following description, well known functions or constructions are not described in detail since they would obscure the description with unnecessary detail.

It is an objective of the present invention to provide the use of Labisia pumila, also known as Marantodes pumilum plant extract for the manufacture of a topical medicament for the treatment of a variety of wounds such as incision, abrasion, avulsion, laceration and puncture. The topical medicament may be in the form of an ointment, a wound bandage or a gauze. An embodiment of the present invention relates to the use of Labisia plant extract in the form of an ointment. The Labisia extract ointment may be formed using the method as illustrated in FIG. 1. Initially, Labisia plant material is dried as in step 101. The plant material includes leaves, stems and roots of the Labisia plant. The Labisia plant material is cleaned and dried at a temperature of approximately 40°C for a period of approximately 3 days.

Thereon, the dried plant material is ground as in step 102. Preferably, the dried plant material is ground into powder form in order to facilitate the subsequent water extraction process. Next, an extract is obtained from the ground plant material using water extraction as in step 103. Preferably, the step of obtaining an extract from the ground plant material using water extraction is conducted at a temperature of 50°C to 70°C for a period of approximately 2 hours under continuous stirring. Preferably, the weight ratio of said ground plant material to said water ranges between 1 : 10 to 1 : 12. Said water can either be distilled water, deionized water or any other purified water. Water is chosen as the initial extraction solvent as water is effective in extracting bioactive compounds of Labisia. Most bioactive compounds in Labisia are polar compounds. Water is more effective in extracting polar compounds in comparison to other organic solvents due to its high polarity and short chain. Furthermore, the presence of hydroxyl group in water forms hydrogen bonding with the polar compounds, which further aids the extraction.

Then, the extract is filtered to obtain a filtrate as in step 104.

Thereon, the filtrate is freeze dried to obtain powdered Labisia plant extract as in step 105. Preferably, the filtrate was cooled at a temperature of -80°C for a day prior to being freeze dried. The step of freeze drying is preferably conducted for a period of approximately 4 days. Freeze drying process function to dehydrate the filtrate at low- temperature operating conditions in order to reduce plant extract deterioration by water, microbial activity and chemical reactions.

Finally, the powdered Labisia plant extract is mixed with a carrier substance to obtain Labisia extract ointment as in step 106. The carrier substance acts as a vehicle, excipient and delivery medium for the extract. The carrier substance may be a semi-solid or liquid material. It may be of purified naturally occurring substances, such as aloe vera and olive oil, or of synthetic substances such as Cetomacrogol emulsifying ointment and petrolatum, or any other carrier substances known in the art. Preferably, said carrier substance is compatible with the powdered Labisia plant extract such that a stable and homogeneous mixture may be formed. Preferably, the percentage by weight of said powdered Labisia plant extract to the total weight of the ointment ranges between 0.5 to 3.0 percent.

Optionally, one or more additives and preservatives may be added into the Labisia extract ointment in order to improve its characteristics, such as fragrances, stabilizers, thickeners, emulsifiers, colouring agents, antioxidants, ultraviolet light absorber and absorption promoting or delaying agents.

The following examples are given to demonstrate the preparation of Labisia extract ointment. Ratios or percentages used in the following examples refer to the weight, unless indicated otherwise.

The examples used herein are intended merely to facilitate an understanding of ways in which the embodiments herein may be practiced and to further enable those of skill in the art to practice the embodiments herein. Accordingly, the examples should not be constructed as limiting the scope of the embodiment herein.

Examples of the Experiments are illustrated as follows: Procedures

These examples illustrate the preparation of Labisia extract ointment from Labisia pumila of the var. pumila and var. alata species. In the present experiment, both leaves and roots of the plant were chosen for extraction. However, the leaves and roots were separately extracted in order to examine their individual wound healing capability. Initially, Labisia plant material was cleaned and dried at 40°C for 3 days. The dried plant material was then ground into powder. Then, the powdered plant material was extracted in distilled water at a temperature of 60°C for 2 hours under continuous stirring. The weight ratio of leaves to water was 1 : 13 while the weight ratio of stem-roots to water was 1 : 10, respectively. The extract was then filtered to obtain a filtrate. Thereon, the filtrate was cooled at a temperature of -80°C for a day. Next, the cooled extract was freeze dried for 4 days until a powdered Labisia plant extract was obtained. The powdered Labisia plant extract was then grinded by mortar and pestle to make it into fine powder. Then, the powdered Labisia plant extract was mixed with Cetamacrogol emulsifying ointment at various concentrations in order to determine the most effective formulation.

Wound healing study in rats

72 female Sprague-Dawley rats aged from 3 to 5 months and weighing between 200 to 250g were used for the wound healing study. Throughout the study, the rats were housed in plastic cages at a temperature of 29 ± 3°C under natural day/night cycle. They were fed with commercial food pellets and deionized water ad libitum. The rats were allowed to acclimate to laboratory conditions for a week before undergoing ovariectomy in order to represent oestrogen deficient state in order to illicit phytooestrogen effects of Labisia. After ovariectomy was performed, the rats were kept for a minimum of two weeks to observe estrogen deficiency state. A mixture solution containing 100 mg/ml of ketamine and 20 mg/ml of xylazine at a ratio of 1 : 1 were injected intraperitoneally to anesthetize the rats prior to all surgical procedures. Excision wound model was used to study the wound healing effect of Labisia extract ointment in ovariectomized rats. Briefly, the rats were generally anaesthetized prior to wound creation. The dorsal surface of rats was shaved using a sterilized razor blade and disinfected with 70% ethanol. Four full skin thickness wounds were made bilaterally using a biopsy punch having a 6mm diameter. The dorsal surface was chosen for this experiment to preclude the rats from biting and stretching the wound area. Thereon, the rats were randomly divided into 9 groups, each containing 8 rats. The rat groups were labelled SH for sham operated rats, OC for ovariectomized rats acting as the negative control, OV for ovariectomized rats treated with the carrier substance Cetomacrogol emulsifying ointment, OF for ovariectomized rats treated with flavine dressing, OE for ovariectomized rats treated with estrogens, PL for ovariectomized rats treated with 1 % of Labisia var. pumila leaf extract ointment, PR for ovariectomized rats treated with 1 % of Labisia var. pumila root extract ointment, AL for ovariectomized rats treated with 2% of Labisia var. alata leaf extract ointment and AR for ovariectomized rats treated with 2% of Labisia var. alata root extract ointment. The wound dressings were changed daily, starting from the wound induction until complete healing. The parameters studied were percentage of wound contraction and the time to complete wound closure.

Wound contraction was measured according to clock method using a digital caliper in μηι. The wound area measurements were conducted on selected days until complete closure of the wound. The results were expressed in percentage of wound healing, %, calculated using the equation below;

[(4) - A_t)

Percentage of wound healing, % = x 100% where A₀ is the initial wound area size and A_t is the wound size after time interval t. Relevant descriptive analysis such as frequency, mean and standard deviation and parametric inferential analysis were performed to a statistically significant level below 5 %.

Photographs of wounds treated with Labisia extract ointment were taken on post-wounding day 2, 5, 8 and daily after until complete wound closure.

Results and Discussion

The rats were treated with Labisia extract ointment containing various percentages of leaf and root extract to determine the most efficient concentration and source for the Labisia extract. The percentages of wound healing for rats treated with Labisia var. pumila extract ointment are shown in TABLE 1.

TABLE 1

TABLE 2 shows the percentages of wound healing for rats treated with Labisia var. alata extract ointment on a daily basis. TABLE 2

It is noted that for Labisia pumila var pumila, both leaf and root extract demonstrated best wound healing result with 1 % concentration with complete wound healing at Day 9 as seen in TABLE 1. Labisia pumila var alata leaf and root extract demonstrated a marginally higher best concentration for wound healing at 2% with complete wound closure noted at Day 9 as well. It can be concluded that the best concentration for wound healing using Labisia pumila var pumila is 1 % while for Labisia pumila var alata is 2%. Photographic representations of rat wounds treated with Labisia extract ointment were captured on post-wounding day 9 and are shown in FIG. 2.

Statistical comparison on the time taken for complete healing of wound between all experimental rat groups were shown in FIG. 3 and Table 3. The data are given as mean ± standard error, S.E, for eight animals in each group. Referring now to FIG. 3, statistically significant results at P<0.001 in comparison to the control group are indicated as with an "*" while statistically significant result at P<0.05 are indicated with an "a". No significant difference was observed among control groups, SH, OC and OV. Control groups, OF and OE, showed slower healing in comparison to control groups, SH, OC and OV. However, rat groups treated with Labisia extract ointment dressings, PL, PR, AL and AR, demonstrated significantly faster healing compared to at least 3 control groups with PR group demonstrated significantly faster healing compared to all control groups. Rats treated to Labisia extract ointment dressings achieved complete healing on post-wounding day 9. Table 3 further compares the differences in complete wound healing days between treated and non-treated groups whereby significant differences between groups are indicated as same alphabet.

TABLE 3

* Differences between groups were calculated using One-way ANOVA with Scheffe post-hoc analysis conducted. Same alphabet indicates significant differences between treated groups and non-treated groups, p-value was set at 0.05.

FIG. 4 shows a graph representing the percentage of wound contraction for all experimental rat groups. The mean percentage of wound contraction was determined on post-wounding day 2, 5, 8, 9, 10, 11 , 12 and 13. At the early stage of wound contraction, post-wounding day 2, the healing rate for all Labisia extract ointment treated groups were less than the control groups with the exception of OV group. From post-wounding day 5 onwards, wound-healing activity of Labisia extract ointment treated rats increased significantly when compared to non-treated groups. The results showed that all treated groups healed approximately at day 9. The process of wound contraction which is the reduction of the wound surface area size depends on the ability to repair damaged tissue and angiogenesis processes, the type and extent of tissue damage and the general condition of the tissue itself. Results obtained from graphical presentation for wounds treated with Labisia extract showed rapid wound contractions. These results suggest that Labisia extract may have a beneficial influence on cells surrounding wound scab. Surrounding cells function to interact with the scab in order to accelerate the wound healing process. As a conclusion, Labisia extract have the potential to be used in the treatment of wounds.

A further embodiment of the present invention relates to a wound bandage comprising an adhesive member adapted for application to a person's skin and a pad on one side of the adhesive member, characterized in that said pad is impregnated with powdered Labisia plant extract. Preferably, said powdered Labisia plant extract is in an amount of 0.5 to 5.0 percentage by weight of said pad. The powdered Labisia plant extract may be impregnated into the pad by any impregnation methods known in the art. It will be readily apparent to those skilled in the art that the powdered Labisia plant extract may be impregnated along with a substantial amount of antiseptic, disinfectant, antibiotic or the like which is suitable for the treatment of wounds or the like in order to reduce the possibility of infection, sepsis, or putrefaction.

In a further aspect, the present invention relates to a gauze comprising a dry textile material impregnated with powdered Labisia plant extract. Preferably, said powdered Labisia plant extract is in an amount of 0.5 to 5.0 percentage by weight of said textile material. The powdered Labisia plant extract may be impregnated into the textile material by any impregnation methods known in the art. It will be readily apparent to those skilled in the art that the powdered Labisia plant extract may be impregnated along with a substantial amount of antiseptic, disinfectant, antibiotic or the like which is suitable for the treatment of wounds or the like in order to reduce the possibility of infection, sepsis, or putrefaction.

While embodiments of the invention have been illustrated and described, it is not intended that these embodiments illustrate and describe all possible forms of the invention. Rather, the words used in the specifications are words of description rather than limitation and various changes may be made without departing from the scope

Claims

1. Use of Labisia or Marantodes plant extract in the manufacture of a medicament for use in the treatment of wounds.

The use of the Labisia or Marantodes plant extract as claimed in claim 1 , wherein said Labisia plant extract is in an amount of 0.5 to 3.0 percentage by weight of the medicament.

A wound bandage comprising an adhesive member adapted for application to a person's skin and a pad on one side of the adhesive member, characterized in that said pad is impregnated with powdered Labisia or Marantodes plant extract.

The wound bandage as claimed in claim 3, wherein said powdered Labisia or Marantodes plant extract is in an amount of 0.5 to 5.0 percentage by weight of said pad.

A gauze comprising a dry textile material, characterized in that said textile material is impregnated with powdered Labisia or Marantodes plant extract.

The gauze as claimed in claim 5, wherein said powdered Labisia or Marantodes plant extract is in an amount of 0.5 to 5.0 percentage by weight of said textile material.