WO2006075932A2

WO2006075932A2 - Remedy to accelerate healing of wound and burn

Info

Publication number: WO2006075932A2
Application number: PCT/RU2005/000670
Authority: WO
Inventors: Nina Sergeevna Demina; Sergei Vasilyevich Lysenko; Eugenie Borisovich Shustov
Original assignee: Otkrytoe Akzionernoe Obschestvo 'sistema Venture'
Priority date: 2005-01-13
Filing date: 2005-12-27
Publication date: 2006-07-20
Also published as: RU2005100533A; WO2006075932A3

Abstract

The present invention relates to a medicine, in particular to remedies for the external application, and can be used to accelerate healing of infected wounds and burns, trophic ulcers, complicated trophic defeats of soft tissues at obliterating vascular diseases of finiteness. A remedy contains antiseptic, osmotically active and hydrophilic components, as well as collagenase of microbial origin. It possesses a high anti-inflammatory, bactericidal and recycling effect at treatment of the different diseases. The invention provides a rapid healing process.

Description

REMEDY TOACCELERATE HEALINGOFWOUNDAND BURN

The present invention relates to the field of medicine, in particular to remedies for the external application and can be used to accelerate healing of infected wounds and burns, trophic ulcers, complicated trophic defeats of soft tissues at obliterating vascular diseases of finitenesses. BACKGROUND OF THE INVENTION

There is known application of antiseptic tanks to accelerate healing of infected wounds, for instance, Konkov ointment is performed with bee product dissolved in fatty substance, water solution containing 15 % antiseptic, as which is used ethacridine (Mashkovskiy M.D., Medical products, V.2, M.: Medicine, 1997, 286 p.). Lack of the ethacridine antiseptic is weak influence on both anaerobic and gram-negative micro flora and the microscopic fungi often infecting wound and burn surfaces as well as having an painting effect on tissues.

There are known ways of treatment of not healing wounds for a long time by means of the preparations containing enzymes, for example, Iruxol ointment (M.A. Trunin, A.S.Yemelyanov "Application of Iruxol in surgical clinic". M. "Surgery", 1982, N 8, p. 88), enzyme tripsin (P.I.Tolstyh and etc. "Influence of biologically active bandages for wounds on current of wound process", M. "The bulletin of surgery", 1987, N 3, p.57). However usage of fermental preparations promotes only to clarification of wounds from necrotic tissues, fibrinogenous strike, but does not influence on processes of regeneration and epithehzation of wounds. One of the lacks of Iruxol is that as an antimicrobial component it contains a chloramphenicol (synthomycin, levomicetin) rendering but only bacteriostatic action not bactericidal and does not render influence on fungoid microflora.

At the same time if the wound surface does not contain a significant amount necrotic tissues and a fibrinogenous strike, Iruxol practically does not

3AMEIMIOmHH JIHCT (πPABHJIO 26) influence on its clarification from purulent separated and the subsequent regeneration and epithelization.

In such cases it is reasonable to use of osmotically active components, which effectively promote washing away from wound surface tissues of microorganisms, bacterial toxins, products of disintegration of tissues and the antiinflammatory biologically active substances formed in a wound process zone. Usage of a hypertonic salt solution for washing of wounds and impregnation of bandages is most known in this context. However, if the bandage containing a hypertonic solution dries up bacterial and endotoxins being direct contact with a wound surface in high concentration sharply brake processes of regeneration and epithelization of wounds. The patent RU 2240780 of 05.09.2003 discloses a remedy ointment for treatment of the infected wounds, which contain antiseptics iodine (1 - 2 %) and a hydrophilic basis - lanolin (17 - 20 %). Additional components of this remedy are benzocaine, beeswax, dimexid, fir oil, and vaseline as ointment bases. Lack of such means is a high frequency of irritating action of iodine (iodism) down to development of chemical burns and allergic reactions on dimexid, and also absence of the components, hydrolizing necrotic tissues and deleting pus and exudation. According to antimicrobial activity spectrum the iodine possesses insufficiently expressed action concerning anaerobic flora and microorganisms, which are deeply being in tissues and nooks. Besides, iodine is capable to brake formation granulation tissues and epithelization of wound surface at some people.

The widely employed surface-active antiseptics are Myramistin and chlorhexidine, which are used for healing of wound and burn surfaces, treatment of the infectious processes on mucous membranes. For example, patent RU 2185155 of 27.04.2000 describes a wound-healing glue containing Myramistin and patent RU2174847 of 01.03.2000 describes a bandage for treatment of the wounds containing chlorhexidine.

3AMEIMIOmHH JIHCT (πPABHJIO 26) Remedy for treatment of burns containing surface-active antiseptics Myramistin and hydrophilic gel basis (patent RU 2185157 of 26.06.2000) is selected as the prototype. However, effectively counteracting development in a wound of pathogenic microflora and supporting necessary for reparation processes hydrophilicity of tissues it does not render direct influence on clarification of a wound from necrotic tissues and endotoxins, as well as on epithelization of wound surface. ^:

SUMMARY OF THE INVENTION

A primary aim of the present invention is to create a multifunctional versatile remedy for accelerating the healing processes, which is applicable to suppress growth of as much as possible wide spectrum of pathogenic microflora (including anaerobic and fungoid), to delete exudation and necrotic tissues from a wound, to carry out their fast healing, to support tissue hydrophilicity not rendering thus allergic and resoφtive reactions. The claimed aim is achieved by that the present remedy includes a surface- active antiseptic, osmotically active component, hydrophilic component and microbial origin collagenase.

Hlorhexidine or Myramistin can be used as a surface-active component, proxanol - as osmotically active component, as a hydrophilic component - lanolin or others hydrophilic components of ointment bases resolved to application in a pharmaceutical practice. Microbial origin collagenase produced from non-pathogenic strain Streptomyces lavendulae (patent RU 2075219 of 23.03.1995) can be used as a fermental component of the declared remedy. Antioxidants, antihypoxants, stimulators of epithelization and regeneration of tissues strengthening the separate parties of its pharmacological activity compatible to the basic components can be entered into structure of a compounding in addition. Any resolved to application in the pharmaceutical practice ointment and gel bases, as well as neutral creams can be applied as a basis.

3AMEIMIOmHH JIHCT (IIPABHJIO 26) EXAMPLES

A studying of healing process of the declared remedy has been lead in comparison with use of the prototype.

A research was carried out with white non-inbred male mice by weight 20-30 g on the model of flat aseptic skin wound. The flap has been cut out at animals in sterile conditions under narcosis (sodium ethaminal) in interscapular region of back after preliminary removal of hair so that the oval wound area should be 80-100 mm². A comparative studying; of wound healing dynamics has been carried out with help of visual supervision and planimetric definition of the wound area has been carried out by means of overlay grid within 2-3 weeks daily estimating the state of the wound edges, wound secretions and formation of the scar. The treatment of animals began at once after the surgery: declared remedy hydrophilic ointment has been put on the wound. There was used the remedies as follows: Test-1 - analogue of the prototype (Myramistin 0,05 % on the lanolin- vaseline hydrophilic ointment basis);

Test-2 - the lanolin-vaseline hydrophilic ointment basis (the percentage of lanolin - 20 %); Test -3 - ointment "Iruxol".

Table 1 shows results of healing activity of declared remedy. This table demonstrates that the present remedy authentically accelerates wound healing since the third day in comparison with both hydrophilic ointment basis and antiseptic ointment Myramistin and ointment "Iruxol" containing collagenase.

Non-inbred white male rats by weight 300 - 320 g have been used at research on model of the infected wound. Hair were cut out from the back cervical area of rats under the general ether narcosis and the flap has been cut out with hypodermic cellular tissue and superficial fascia at aseptic conditions. The teflon ring (diameter of 8 mm) covered from above the punched film was

3AMEIMIOmHH JIHCT (πPABHJIO 26) inserted into the wound. The wound edges and bottom have been injured by gear clip additionally. Suspension of daily culture of pathogenic microorganism Staphylococcus aureus (golden staphylococcus) in amounts of 0,5 ml approximately corresponding concentration 1,5 x 10⁹ microbial cells/ml of suspension was entered into the wound surface inside of the ring. Rings were removed after 2 days and there was spent a standard primary surgical processing of wound and then there was began a treatment by declared remedy. There was used the remedies as follows: Test-1 - analogue of the prototype (Myramistin 0,05 % on the lanolin- vaseline hydrophilic ointment basis);

Test-2 - the lanolin-vaseline hydrophilic ointment basis (the percentage of lanolin - 20 %); Test -3 - ointment "Iruxor. Each testing group has been contained 8 animals.

A cytological research of healing process has been used as the method to estimate speed and completeness of wound treatment. For this purpose there was made an impression smear from the wound surface by the touch of subject glass to the wound preliminary cleared of pus and fibrin film by the hydrogen peroxide solution on the third day after surgical processing of the wound. The smear was dried on air then it was fixed by methyl spirit and after that it was painted by Romanovskomu-Gimze method. There were counted up at least 400 cells in one smear: neutrophilous leukocytes, mononuclear cells, macrophages, fibroblasts and then there was calculated a percentage. Table 2 shows the research results.

The analysis of Table 2 describes an improvement of the infected wound healing process under influence of both Myramistin and Iruxol, as well as declared remedy. Thus, there is noted a lowest level of the neutrophilous leukocytes reflecting reaction of the wound tissue on microbial contamination for declared remedy (authentic differences from both Test-2

3AMEIMIOmHH JIHCT (πPABHJIO 26) and Test-3). Differences from Myramistin level by this parameter are not authentic because of equal content of the antiseptic in both the present remedy and Test-1. However, there is certain strengthening of activity due to other components of remedy also. Parameters of the fibroblast level reflecting activity of wound healing process are maximal also for declared remedy and authentically differ from all kinds of the tests. A research of qualitative characteristics of formed during wound healing scar was carried out on model of the skin wound. For this purpose after preliminary cutting hair in interscapular areas of back of the laboratory animals (non-inbred white male rats by weight 300 - 310 g) under the general ether narcosis there has been made 2 linear cuts through all skin layers by 1 cm length: one - on the right site and one - on the left site of backbone approximately at 1,5 - 2 cm distance from it. One interrupted stitch has been imposed on the wound by sterile catgut suture. The present remedy has been laid on one linear wound, the testing preparation - on another. There was used the remedies as follows:

Test-1 - analogue of the prototype (Myramistin 0,05 % on the lanolin- vaseline hydrophilic ointment basis);

There were 15 animals in the research group and 5 animals - in the testing group.

After surgery on 13-th day there were taken slices of tissue from wound scar zone for histological research. They was fixed in Karnua liquid, then there were produced step paraffin cuts and were painted by hematoxylin and eosin. The analysis of Table 3 demonstrates that application of declared remedy promotes formation of a mature connecting tissue without predisposition to

JIHCT (πPABHJIO 26) keloid scarring. Both declared remedy and Myramistin promote faster transition of fibroblasts into febricities on hydrophilic ointment basis. Thus, the carried out researches have confirmed that the declared remedy exceeds preparations of comparison at ability to accelerate healing both infected and non-infected wounds, promotes faster and qualitative scarring of them and epithelialization without increase of risk of keloid scar appearance.

3AMEIMIOmHH JIHCT (IIPABHJIO 26) Table 1

Testing the healing of aseptic flat wounds by Remedy

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Table 1 (continuation)

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Table 2

Testing the healing of infected wounds by remedies (microscopy of smear from the wounded surface, group mean, n=7)

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Note:* - differences with respect to Test-2 are known for certain, p< 0,05 (criterion of exact Fisher's method) # - differences with respect to Test-3 are known for certain, p< 0,05 (criterion of exact Fisher's method)

Table 3

Qualitative indices of the healing wounds (model of the linear, histological indices)

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Note: * - differences with respect to Test-2 are known for certain, p< 0,05

(criterion of exact Fisher's method)

Claims

1. A remedy to accelerate healing of wound and burn comprising an antiseptic, and characterized in that it additionally comprises both an osmotically active hydrophilic component and a collagenase of microbial origin with activity 3 - 10 units in 100 grams by weight.

2. A remedy according to claim 1, wherein the antiseptic is Myramistin representing 0,1 to 0,5 % by weight, the osmotically active component is a proxanol representing 20 to 30 % by weight, hydrophilic component and ointment base is a polyethilenoxide 400 up to 100 % by weight.

3. A remedy according to claim 2, wherein the antiseptic is a glorgecidine bigluconat representing 0,1 to 0,5 % by weight.

3AMEIMIOmHH JIHCT (IIPABHJIO 26)