WO2018070956A2

WO2018070956A2 - Wound and burn material with herbal agent

Info

Publication number: WO2018070956A2
Application number: PCT/TR2017/050373
Authority: WO
Inventors: Asligul KURT; Erdal ERDOGAN
Original assignee: Doku Biyoteknoloji Limited Sirketi
Priority date: 2016-08-11
Filing date: 2017-08-08
Publication date: 2018-04-19
Also published as: WO2018070956A3; GB2566630A8; GB2566630A; GB201820503D0

Abstract

The invention relates to a wound and burn material which is intended to be used in healthcare field, promotes healing of acute and chronic wounds, has two bioabsorbable, gelatin matrix based layers with clotting effect, and contains cinnamon extract, tannic acid and a crosslinking agent, and to a method for production of the mentioned material.

Description

WOUND AND BURN MATERIAL WITH HERBAL AGENT

Technical Field The invention relates to a wound and burn material which is intended to be used in healthcare field, promotes healing of acute and chronic wounds and contains a bioabsorbable herbal agent with clotting effect, and to a method for production of the mentioned material.

Essentially, the invention relates to a gelatin (hydrocolloid polymeric matrix) based wound-burn material comprising cinnamon extract, which is obtained from dried barks of Cinnamomum verum tree and has collagen synthesis triggering function, crosslinking agent and clotting tannic acid, and a method for its production.

Present State of the Art Traditional or modern wound dressings and similar materials are effective in terms of wound recovery and reducing the risk of infection, however, they are known to be inadequate for regenerating the lost tissue and regenerating the dermis layer lost in burn. Today the most helpful wound care matrix materials are developed from hydrocollids such as gelatin, collagen, silk peptides, fibrin, poly(glycolicacid)(PGA) and poly(lactic acid) (PLA).

Many products marketed by the pharmaceutical industry such as creams, salves, pomades are mostly effective in preventing an infection of the present wound and they are rather inadequate for ensuring tissue integrity. On the other hand, ensuring tissue integrity and various bioactive molecules, glycoprotein and peptide structures, a framework which will prompt cells to attach together, and suitable humidity-temperature conditions are highly important in wound-burn healing process.

Different wound care products produced from natural polymers or various synthetic materials are commercially available. However, they fail to deliver required results since they are not produced in a three dimensional structure and therefore they are ineffective for use with wounds which require depth. On the other hand, there are disadvantages associated with synthetic materials such as not being bioabsorbable, having risk of allergy and necessity to be removed from the wound. This results in damaging the granulation tissue, which consists of the first phase of the wound healing process, and even in some cases causes a recurring bleeding of the wound.

Grafts, which are another example of wound dressing materials, are particularly preferred in burn and acute laceration related tissue losses. However, the mentioned material is not preferred since surgical intervention is required in the treatment, for which the mentioned material is used, and a matched donor is required. Moreover, it is known that the transplant tissue may also be damaged, immune reactions due to the graft and related infections can be seen in the patient. Primary treatment step in acute lacerations is stopping the bleeding and starting the healing phase by achieving a strong clotting. Existing products are only aimed towards achieving the clotting and commercially available in liquid form sprays, powder form structures and various medication forms such as fibrin adhesives. While these products have no wound healing function, they cannot be used in treatment of chronic open wounds.

Therefore, based on the aforementioned disadvantages and inadequacy of the existing products in the relevant area, an innovation is necessary in the relevant technical field.

A Brief Description of the Invention Present invention relates to a wound and burn material with a herbal agent which meets the aforementioned requirements, overcomes all disadvantages and provides some further advantages.

Primary purpose of the invention is to produce an effective and combination wound and burn material for rapidly providing tissue integrity in acute and chronic wounds. A further purpose of the invention is to develop a wound material comprising a tree bark (cinnamon) extract which promotes regeneration by triggering collagen synthesis in both layers, to overcome the slow regeneration which is the most important problem encountered during wound healing. Most of the wound dressings comprise growth factors to increase regeneration. On the other hand, these growth factors increase cancer risk while prompting regeneration. It has been observed that the mentioned tree bark (cinnamon) extract provides a substantial increase in regeneration by supporting the cell activity, when applied in a gelatin matrix. Employing the regenerative and antimicrobial features of this extract in a biomaterial is the innovative aspect of the invention.

A further purpose of the invention is develop a wound healing biomaterial comprising a tree bark (cinnamon) which contributes to the synthesis of intermediate molecules required in various stages of cell activities (cell growth, aging, differentiation, multiplication, combination, migration and programmed cell death). Furthermore, there are scientific studies which report the related wound healing biomaterial to contribute in reducing the cellular stress and preventing the spread of cancer cells.

The invention may also provide cosmetic expectations of the patients especially in burn cases, in addition to the wound healing features of the cinnamon extract. Cinnamon extract of the invention contributes to the restoration of the wound cosmetically by reducing scar tissue formation due to degeneration of the skin elasticity and lack of humidity, and this also supports the rehabilitation process in psychological aspects.

A further purpose of the invention is to develop a wound-burn material, intended to be used also with acute wounds, which comprises "tannic acid", which is also known as "tannin", which is available in plants and used in medicine since old times for its vascular and mucosa contracting (clotting mechanism) effect.

A further purpose of the invention is to develop a three dimensional and porous wound healing biomaterial by using a herbal crosslinking (genipin) agent, without causing high toxicity side effects.

A further purpose of the invention is to provide a more effective wound-burn material production process, without causing temperature sensitive repairing and regenerative molecules to be denaturized, by employing a freeze drying process.

A further purpose of the invention is to develop a fully bioabsorbable wound-burn material, which comprises 2 layers, includes key features required in critical steps of wound healing and having a synergic effect at the same time.

Intrinsic and characteristic features of the invention and all its advantages will be readily understood with reference to the detailed description. Therefore, it is necessary to take these figures and the detailed description into consideration during assessment. Detailed Description of the Invention

In this detailed description, a wound and burn material with a herbal agent is only presented only for better appreciation of the subject matter and it does not impose any limiting effects. One of the agents used in production process of the wound and burn material with herbal agent is cinnamon. Cinnamon extract, obtained from dried barks of Cinnamomum verum tree, promotes synthesis of proteins such as collagen and elastin by stimulating fibroblast cells, which are main skin cells. Therefore it supports scabbing of the open wound while at the same time being incorporated in the cell membrane structure. Cinnamon extract, with its antibacterial and antifungal features, kills bacteria, helps preventing growth of resistant fungi and therefore development of infection is prevented. Cinnamon extract is rich in iron and calcium; and it is effective in clotting and enhancing the immune system. In addition to the abovementioned features, its usage in wound-burn treatment provides an important innovation to the technical field since it is an affordable plant extract.

The other active agent used in the composition provided in the invention is tannic acid. Tannic acid supports the clotting mechanism by contracting the blood vessels and reducing the blood flow with its (anti-vasodilator) effect. It is antimicrobial and functions to protect the wound against infections. In addition, its antioxidant and antimutagenic features are also among the most important properties of tannic acid. Therefore it is advantageous in preventing chronic wounds becoming cancerous.

In the wound and burn material with herbal agent, genipin is used as a crosslinking agent. Genipin, which has much lower cytotoxicity compared to other crosslinking agents such as gluteraldehide, is more advantageous compared to glutarealdehide since it shows longer lasting stability. Therefore it provides a more stable 3 dimensional structure and presents high biocompatibility. In the wound and burn material with herbal agent according to the invention; genipin, gluteraldehide or aldehyde derivatives can be used as the relevant crosslinking agent.

Gelatin (hydrocolloid polymeric matrix) is another raw material that is present in the composition of the woung and burn material with herbal agent. It is an essential matrix element used in various biomaterials, in chemistry, pharmaceutics and food industry. The mentioned gelatin can be of animal or plant origin. Its purpose of use in the invention is controlled transfer of the relevant active agents inside the tissue and providing a foam that acts as a framework to help the cells attach together. Since gelatin has a polypeptide form, it also contributes to wound healing by being absorbed by body.

Table 1 shows the raw materials used in the wound and burn material with herbal agent and ratios of their amounts. Following the basic review of the production steps, it is disclosed clearly in reference to the detailed description without being limiting in any way.

Wound and burn material production process basically involves the following steps:

□ Obtaining C.verum tree bark extracts and alcohol (methanol or ethanol),

□ Preparation of gelatin solution,

□ Adding C.verum tree bark extracts (cinnamon) to gelatin solution,

□ Splitting the gelatin and cinnamon extract containing solution into 2 containers,

□ Separately preparing the genipin solution,

□ Adding tannic acid into one of the splitted containers,

□ Obtaining foam structures from splitted 2 containers,

□ Adding previously prepared genipin solution into foam structures in splitted 2 containers and thus enabling crosslinking of foam structures to take place,

□ Obtaining a semisolid consistency by freezing the crosslinked foam structures at -20 °C,

□ Physically combining semisolid structures from 2 splitted containers and solidifying them as a 2 layer form by freezing at -80 °C, □ Drying the solidified 2 layer structure,

□ Obtaining a 2 layer, 3 dimensional porous sponge structure.

Wound and burn material production process starts with obtaining Cinnamomum verum tree barks (cinnamon), alcohol extracts. After powdering the obtained cinnamon by grinding, this cinnamon powder is added into alcohol with 1 /6 (w/v) ratio and shaken for 24 hours at 25 °C. This shaken mixture is distilled with filter paper and the remaining colorful liquid evaporated using an evaporator. At the end, total extract is obtained.

A 5 g of the pharmaceutical gelatin in the powder form is added into 100 ml purified water. Gelatin is dissolved inside a magnetic stirrer for 20 minutes at 37 °C and 250 rpm and homogenized. Finally, a 5% (w/v) gelatin solution is obtained.

About 10 mg of the extract, which has been shaken with alcohol, filtered and liquid was evaporated from, is added into 100 ml gelatin solution. Cinnamon extract is dissolved and homogenously distributed at 37 °C and 250 rpm for 10 minutes. At the end, a gelatin solution containing %0.01 (w/v) is obtained. Obtained 100 ml solution is splitted into 2 separate 50 ml containers using a graduated cylinder.

In a separate container, a total of 6 g pure genipin dust is added into 100 ml purified water at room temperature an distributed homogenously by stirring at 250 rpm for 10 minutes. Solution is maintained between +2 °C - +12 °C until the time of use and a solution with 6% (w/v) concentration is obtained.

0,5 g tannic acid powder is added into only one of the solutions containing 50 ml gelatin and cinnamon extract and homogenously dissolved by stirring at 37 °C and 250 rpm for 10 minutes in a magnetic stirrer. Then the tannic acid concentration becomes 1 % (w/v).

Solutions prepared inside 2 separate containers are, while still keeping them in separate containers, are stirred at room temperature and at 1400 rmp for 15 minutes until foam structures are obtained.

After obtaining foam structures inside 2 different containers, 10 ml of 6% genipin solutions is added into both of the containers by the help of a pipette. A genipin concentration of 1 % w/v is reached in both of the containers. Stirring is continued for 10 more minutes at 37 °C and 1400 rpm to ensure crosslinking of foam structures. Foams that have been crosslinked inside 2 separate containers are poured into moulds separately. For the foams to reach a semisolid consistency, all the moulds are left to cooling inside a deep freezer at -20 °C for 30 minutes.

After the cooling process, semisolid foam structure inside either one of the moulds are carefully removed from the mould and placed on the other semisolid foam structure. To allow this 2 layer structure to attach together, it is left to solidify inside a deep freezer at -80 °C for 4 hours.

After completion of the solidification process, frozen, 2 layer foam structure is placed into a freeze dryer device for drying process. Drying process is conducted under 20Pa pressure for 18 hours.

The resulting material from the device is the final product which has a spongy form, is porous and has a 2 layer structure.

When the layered hydrocolloid polymer mentioned in the invention absorbs exudate, tannic acid in the first layer diffuses into the tissue. Depending on the condition of the exudate and at most in 12 hours, the first layer dissolves and the second layer comes into contact with the wound. Fibroblast cells are excited and collagen synthesis is triggered since both layers contain cinnamon extract, which supports cell proliferation, has a regulating effect on apoptosis mechanisms, prevents excessive drying of the wound and provides protection against infection with its antimicrobial quality. Therefore, the condition of the wound is improved into a healing phase and the wound heals quickly.

The invention, which achieves an improvement in the normal healing duration by 1 /4 as a result of the conducted studies, is suitable for use with both acute and chronic wound cases. The drying process mentioned in the invention is based on absorption of the water molecules, which are present in the frozen material, under high pressure. This process is one of the required intermediate steps that is necessary to achieve the spongy material provided by the invention. The aforementioned 3 dimensional porous structure provides the physical structure required for the cells to attach together and helps a controlled emission of the active agents to the area of the wound by acting as a framework. Wound and burn material with herbal agent is placed onto any kind of open wound without an infection or right after an acute injury with bleeding, on the surface where the skin integrity is damaged, in a manner where the tannic acid containing layer of the invention comes into contact first. Especially in cases with acute bleeding, layer that contains tannic acid will support the clotting process by contracting the blood vessels and reducing the blood flow, and later upon contact with the other active agent, tissue healing process will start. In chronic open wound cases again the tannic acid containing layer will enable the cells to attach together by contracting the tissue to restore the tissue integrity and with the other components of the invention melting on the wound surface, a synergic effect will be observed.

The composition of the invention can be used in acute injuries where a clotting effect is required and any kind of wounds when a rapid tissue healing is expected and not infected wounds. At the same time, its bioabsorbability enables it to be absorbed entirely by the wound in at most 24 hours. After applying the wound and burn dressing to the wound, it turns into a gel form by mixing with the wound fluid and entirely covers the wound surface; and also by covering the open nerve endings, it shows pain relieving effect.

□ It is also possible to obtain a spray based product with liquid form instead of the product with spongy form. A wound and burn material with liquid for which is obtained by placing a solution containing cinnamon extract, gelatin and tannic acid and another solution containing only gelatin and cinnamon extract into spray containers without mixing can be applied by spraying. To be used without mixing with the other solution, tannic acid containing gelatin-cinnamon solution should be transferred to the first phase storage place of the spray bottle. Gelatin- cinnamon solution that does not contain tannic acid should be transferred to the second phase storage place of the spray bottle to be used without mixing with the other solution. In this way, foam production and crosslinking agent usage is not required. On the other hand, it is known that in case of wounds where the wound surface is required to be covered with gelling or wounds with depth, spongy 3 dimensional structure is known to yield better results. Product with liquid form is more suitable for practical use in terms of handling and storage convenience.

Claims

A wound and burn material with herbal agent which is intended to be used in healthcare and supports healing of acute and chronic wounds, has clotting effect, contains gelatin, wherein it has cinnamon extract, tannic acid and crosslinking agent in its content.

A wound and burn material with herbal agent according to claim 1 , wherein it has cinnamon extract between 0.001 -0.1 % range, tannic acid between 0.1 - 10 % range, 0.07 - 3 % genipin, 0.1 - 10 % gelatin in its content.

A wound and burn material with herbal agent according to claim 1 , wherein the mentioned gelatin is of animal and/or plant origin.

A wound and burn material with herbal agent according to claim 1 , wherein the mentioned crosslinking agent is genipin, gluraldehyde or aldehyde derivative.

A method for production of the wound and burn material with herbal agent which is intended to be used in healthcare, wherein it involves following process steps:

□ Preparation of crosslinking agent solution,

□ Adding cinnamon extract to the gelatin solution,

□ Splitting the cinnamon extract-gelatin solution into two containers,

□ Adding tannic acid into one of the solutions split into two containers,

□ Producing a 3 dimensional 2 layer spongy solid product or a liquid based spray product from mentioned solutions of 2 separate containers.

A method for production of the wound and burn material with herbal agent according to claim 5, wherein it involves following steps for obtaining a 3 dimensional 2 layer spongy solid product:

□ Obtaining foam structures inside the mentioned two containers,

□ Adding the crosslinking solution to mentioned foam structures to enable crosslinking,

□ Obtaining a semisolid consistency by freezing the crosslinked foam structures and physically combining them in a 2 layer form,

□ Drying the solidified 2 layer structure. A method for production of the wound and burn material with herbal agent according to claim 5, wherein it involves following steps for obtaining a spray based liquid product:

□ Transferring the tannic acid containing gelatin-cinnamon solution to a first phase storage place of the spray bottle to be used without mixing with the other solution,

□ Transferring the gelatin-cinnamon solution that does not contain tannic acid to a second phase storage place of the spray bottle to be used without mixing with the other solution.