CLIAMS:We Claim,
1. A stabilized wound healing composition without the side effect of burning sensation, the claimed composition comprises of therapeutically effective amounts of neutral Collagen, Gentamicin, Aloevera, Preservative and pharmaceutically acceptable carrier wherein the said composition is capable of contacting the wound or to an open ulcer and thereby the collagen breaks down and the Gentamicin is released into the wound or ulcer.

2. The stabilized wound healing composition as claimed in claim 1 wherein the said neutral Collagen, Gentamicin, Aloevera, Preservative and pharmaceutically acceptable carrier being present therein in a proportion producing a synergistic activity with regard to the wound healing activity conferred.

3. A stabilized wound healing formulation without the side effect of burning sensation, the claimed formulation comprises of 40 to 65 % of neutral Collagen, 0.1 to 0.2 % of Gentamicin, 35 to 60 % of Aloevera, 0.3 to 0.55 % of Preservative and pharmaceutically acceptable carrier wherein the said formulation is capable of contacting the wound or to an open ulcer and thereby the collagen breaks down and the Gentamicin is released into the wound or ulcer

4. A stabilized wound healing formulation without the side effect of burning sensation, the claimed formulation comprises of 40% of neutral Collagen, 0.1 % of Gentamicin, 60% of Aloevera, 0.5% of Preservative and pharmaceutically acceptable carrier wherein the said formulation is capable of contacting the wound or to an open ulcer and thereby the collagen breaks down and the Gentamicin is released into the wound or ulcer.

5. The said neutral collagen in any of the preceding claims includes neutral succinylated collagen

6. The said neutral succinylated collagen of claim 5 is prepared by a process comprising
a. Harvesting collagen from –Bovine Tendon/Fish bladder/porcine Tendon
b. cutting and mincing collagen to formed minced collagen
c. subjecting the minced collagen to non-ionic detergent treatment, alkaline treatment followed by enzymatic treatment to obtain normal tissue
d. carrying out succinylation reaction of obtained normal tissue in neutral pH medium to form swell neutral succinylated collagen wherein additional succinyl group is added to prevent subsequent cleavage of lysil peptide bonds with trypsin as a result of succinylation and thereby making the modified protein acts as better substrate for proteases.

7. The wound healing composition/formulation in any of the preceding claims, wherein said pharmaceutically acceptable carrier is an ointment, gel, white petrolatum, light mineral oil, or mixtures thereof.
8. The wound healing composition/formulation in any of the preceding claims wherein the said composition /formulation is capable of evenly spreading over the wound/ulcer for enhanced efficacy.
9. The wound healing composition/formulation in any of the preceding claims wherein the said composition /formulation is stable for atleast 24 months without decomposing into liquid or precipitate.

Dated this 22nd day of June 2015
For COLOGENESIS HEALTHCARE PRIVATE LIMITED
By its Patent Agent
Dr.B.Deepa

,TagSPECI:Form 2

THE PATENT ACT, 1970
(39 of 1970)
&
THE PATENT RULES, 2003
COMPLETE SPECIFICATION
(See section 10 and rule 13)

“MEDICATED COLLAGEN GEL”

in the name COLOGENESIS HEALTHCARE PRIVATE LIMITED, an Indian national having address at Plot No 51,The Salem Co-Operative Industrial Estate Udayapatti,Salem-636140 Tamil Nadu, INDIA.

The following specification particularly describes the invention and the manner in which it is to be performed
FIELD OF THE INVENTION:

The present invention relates to a wound healing composition. More particularly the present invention relates to a topical therapeutic composition for accelerating wound healing.

BACK GROUND OF THE INVENTION AND PRIOR ART:

A wound is a break or opening in the skin. Wounds in mammalian tissue result in tissue disruption and coagulation of the microvasculature at the wound face. Tissue growth and repair are biologic systems wherein cellular proliferation and angiogenesis occur in the presence of an oxygen gradient. The cellular morphology consists of three distinct zones. The central avascular wound space is oxygen deficient, acidotic and hypercarbic, and has high lactate levels. Adjacent to the wound space is a gradient zone of local anemia (ischemia) which is populated by dividing fibroblasts. Behind the leading zone is an area of active collagen synthesis characterized by mature fibroblasts and numerous newly-formed capillaries. There are reports available in the literature for the availability of various wound healing compositions.
US 5487899 discloses a composition derived from the Aloe plant which when used as an adjuvant for the healing of wounds exhibits increased anti-inflammatory and wound healing activity. The composition, an Aloe vera mucilage at approximately a 1:1 ratio, preferably with Aloe pulp fibers, increased open wound healing activity and increased wound tensile strength by 131% over controls.
US 4,760,131 discloses a soft tissue wound healing composition comprising an aqueous mixture of fibrillar collagen, heparin, and undegranulated platelets or platelet releasate. The composition is applied topically to the wound site in conjunction with means to keep it at the site and hydrated or in the form of an occlusive dressing.
US 5,156,847 discloses a wound-healing composition of the aporphine-derived alkaloid taspine, dissolved in non-aqueous solvent, such as dimethylsulfoxide (DMSO)
US 5,698,228 discloses a wound healing composition comprising a sheet of squid chitin laminated to a layer of fish skin collagen wherein the fish skin collagen is not cross linked.
US 6,046,160 discloses a composition for use in promoting the healing of surface wound, comprising a liquid aqueous suspension of vitamins C and E and collagen type I in admixture with a starch hydrolysate with a DE of less than 35 that exhibits a chemotactic effect on fibroblasts or endothelial cells.
US 7,786,074 provided compositions and methods for use in promoting wound healing and tissue regeneration following tissue injury in a subject. The composition comprising at least one polypeptide consisting of the carboxy-terminal most 4 to 30 contiguous amino acids of an alpha connexin protein or conservative variant thereof, wherein said at least one alpha connexin polypeptide is linked at its amino terminus to a cellular internalization transporter.
From the prior art it is inferred about the availability of various composition for wound healing. However they suffer various drawbacks which include decreased efficiency in healing the wound, instability of the product and also causes burning sensation.
Moreover the collagen gels available in the market are in cream form in the paraffin base which results in uneven spreading of the cream and also the collagen content is very less. Further the gels are highly unstable and convert into liquid or sometimes it precipitates. At present collagen gel is available in acidic pH form which is highly unstable and causes sensation of irritation due to acidic pH. Furthermore the collagen when mixed with any other ingredients tends to get precipitated.
Thus there exists an urgent need in the state of art to formulate a wound healing composition which is devoid of above said drawbacks.

OBJECT OF THE INVENTION:

The main object of the present invention is to formulate a highly stable wound healing composition comprising of neutral Collagen, Gentamicin, Aloevera, Preservative and pharmaceutically acceptable carrier.
Another object of the present invention is to formulate a highly stable wound healing composition comprising of neutral Collagen, Gentamicin, Aloevera, Preservative and pharmaceutically acceptable carrier being present therein in a proportion producing a synergistic activity with regard to the wound healing activity conferred.
Another object of the present invention is to utilize neutral succinylated collagen as neutral collagen in the composition.
Yet another object of the present invention is to formulate a wound healing composition which is devoid of the side effect of burning sensation due to presence of succynilated collagen which swells in neutral pH thereby overcoming the condition of acidic collagen pH.
Yet another object of the present invention is to formulate a wound healing composition which is capable of contacting the wound and thus the collagen breaks down and the Gentamicin is released into the wound or ulcer.
Yet another object of the present invention is to utilize aloevera as a stabilising agent and Natural Healer in the composition.
Yet another object of the present invention is to formulate a wound healing composition which is capable of evenly spreading over the wound for enhanced efficacy.
Further object of the present invention is to formulate the wound healing composition into an ointment, gel, white petrolatum, light mineral oil, or mixtures thereof.

BRIEF DESCRIPTION OF DRAWINGS:

Figure 1 illustrates photographs of a human subject burned and treated with formulation of the present invention to enhance healing, according to one embodiment of the present invention
Figure 2 illustrates photographs of a human subject cut injured and treated with formulation of the present invention to enhance healing, according to one embodiment of the present invention

SUMMARY OF THE INVENTION:

The present invention discloses a stabilized wound healing synergistic composition comprises of therapeutically effective amounts of neutral Collagen, Gentamicin, Aloevera, Preservative and pharmaceutically acceptable carrier. The neutral collagen used in the composition includes neutral succinylated collagen. The composition is capable of contacting the wound or to an open ulcer and thus the collagen breaks down and the Gentamicin is released into the wound or ulcer for effective healing. The wound healing composition of the present invention is devoid of the side effect of burning sensation due to presence of succynilated collagen which swells in neutral pH and thus overcoming the condition of acidic collagen pH.

DETAILED DESCRIPTION OF THE INVENTION:

The present invention relates to a process for the healing of wound using collagen and Gentamicin combination.
The importance of collagen as a biomaterial rests largely on the facts that it is a natural material of low immunogenicity and is therefore recognised by the body as a normal constituent rather than a foreign matter .Collagen has been produced in range of physical forms such as sheets, sponges, powders etc to be utilized in medical process.
Gentamicin is an amino glycoside antibiotic used to treat many type of bacterial infection. It is an bactericidal antibiotics that works by binding the 30s subunit of the bacterial ribosome by interrupting protein synthesis.
The Gentamicin collagen cream when applied on the wound or to an open ulcer the collagen breaksdown and the Gentamicin is released in the ulcers.
Fibrous collagen possesses high tensile strength and its degree of natural cross linking makes it easier to purify without degradation. Tendon collagen is relatively purer and more resistant to decomposition in view of inherent cross-links, which are very firm.
The helical structure of collagen provides its resistance to proteolysis and also masks the antigenic determinants.
For use as a biomaterial ,collagen is recovered in the undenatured form i.e with a little or no damage to the basic rigid triple helical structure. The undenatured collagen in fibril form is obtained by acid,base,salt and enzyme extraction.
Collagen in its nature tends to get precipitated soon when there is change in the temperature and there is also chance of showing burning sensation when the gel is in acidic PH so in order to overcome these issues the collagen is modified by means of adding additional succinyl group to the ends of the protein molecule which is known as succinylation
The tendon is cut into bits and minced thoroughly and is subjected to non-ionic detergent treatment, alkaline treatment followed by enzymatic treatment to obtain normal tissue.
The obtained tissue which gets swells in acidic pH is changed from its form that it gets swell in the neutral pH by means of the process called succinylation reaction during which additional succinyl group is added to it. As a result of succinylation subsequent cleavage of lysil peptide bonds with trypsin is prevented moreover the modified protein acts as better substrate for proteases.
The tissue which is succinylated get swell in the neutral PH and gel is formed to which aloevera and required antibiotic and preservative is added and filled in the tubes for dispatch.
The inventiveness of the invention lies in the following .
• The collagen is succinylated and the nature of the collagen is changed and it is made stable.
• Due to change in its nature the acidic nature is modified which rules out the problem of irritation or burning sensation.
• The collagen gets easily mixed with any ingredients which help in preserving the material in gel form.
• Since the collagen is water based without any greasy material the spreading of layer is very easy.
• When the layer is formed it acts as a barrier to the wound.
• The impregnation or addition of Gentamicin in the gel helps to control the pathogen in the wound.
• The content t of the collagen in the compostion of the present invention is more than 90%.
• There is even spreading of the gel since it is fully water based.
• The stability of the gel is maintained for longer period.
• The gel when mixed with the aloevera gets more stable than any other gel.
• The aloevera which is used as a stabilising agent acts as an Natural Healer also.
In one of the preferred embodiment the present invention shall disclose a stabilized wound healing composition without the side effect of burning sensation. The composition comprises of therapeutically effective amounts of neutral Collagen, Gentamicin, Aloevera, Preservative and pharmaceutically acceptable carrier in which the composition is capable of contacting the wound or to an open ulcer and thereby the collagen breaks down and the Gentamicin is released into the wound or ulcer.
In another preferred embodiment the present invention shall disclose the stabilized wound healing composition in which the neutral Collagen, Gentamicin, Aloevera, Preservative and pharmaceutically acceptable carrier being present therein in a proportion producing a synergistic activity with regard to the wound healing activity conferred.
In another preferred embodiment the present invention shall disclose a stabilized wound healing formulation without the side effect of burning sensation. The formulation comprises of 40 to 65% of neutral Collagen, 0.1 to 0.2 % of Gentamicin, 35 to 60% of Aloevera, 0.3 to 0.55 % of Preservative and pharmaceutically acceptable carrier in which the formulation is capable of contacting the wound or to an open ulcer and thereby the collagen breaks down and the Gentamicin is released into the wound or ulcer.
In another preferred embodiment the present invention shall disclose a stabilized wound healing formulation without the side effect of burning sensation. The formulation comprises of 40% of neutral Collagen, 0.1 % of Gentamicin, 60% of Aloevera, 0.5% of Preservative and pharmaceutically acceptable carrier in which the formulation is capable of contacting the wound or to an open ulcer and thereby the collagen breaks down and the Gentamicin is released into the wound or ulcer.
As per the invention the neutral used in the formulation includes neutral succinylated collagen
In accordance with the invention the neutral succinylated collagen is prepared by a process comprising
a. Harvesting collagen from –Bovine Tendon/Fish bladder/porcine Tendon
b. cutting and mincing collagen to formed minced collagen
c. subjecting the minced collagen to non-ionic detergent treatment, alkaline treatment followed by enzymatic treatment to obtain normal tissue
d. carrying out succinylation reaction of obtained normal tissue in neutral pH medium to form swell neutral succinylated collagen in which additional succinyl group is added to prevent subsequent cleavage of lysil peptide bonds with trypsin as a result of succinylation and thus making the modified protein acts as better substrate for proteases.

According to the invention the pharmaceutically acceptable carrier present in the wound healing composition/formulation can be an ointment, gel, white petrolatum, light mineral oil, or mixtures thereof.
As per the invention the wound healing composition/formulation is capable of evenly spreading over the wound/ulcer for enhanced efficacy.
In accordance with the invention the wound healing composition/formulation is stable for atleast 24 months without decomposing into liquid or precipitate.
Example : 1
The stabilized wound healing composition of the present invention comprises of combination of neutral Collagen, Gentamicin, Aloevera, Preservative and pharmaceutically acceptable carrier.
Example : 2
The stabilized wound healing composition of the present invention comprises of therapeutically effective amounts of neutral Collagen, Gentamicin, Aloevera, Preservative and pharmaceutically acceptable carrier.
Example : 3
The stabilized wound healing composition of the present invention comprises of 40% of neutral Succinylated Collagen, 0.1 % of Gentamicin, 60% of Aloevera, 0.5% of Preservative and pharmaceutically acceptable carrier.
Example : 4
The stabilized wound healing composition of the present invention comprises of 40 to 65% of neutral Succinylated Collagen, 0.1 to 0.2 % of Gentamicin, 35 to 60% of Aloevera, 0.3 to 0.55 % of Preservative and pharmaceutically acceptable carrier.
Example : 5
The stabilized wound healing composition of the present invention comprises of 50% of neutral Succinylated Collagen, 0.125 % of Gentamicin, 50% of Aloevera, 0.23% of Preservative and pharmaceutically acceptable carrier.
Example : 6
The stabilized wound healing composition of the present invention comprises of 60% of neutral Succinylated Collagen, 0.175 % of Gentamicin, 40% of Aloevera, 0.35% of Preservative and pharmaceutically acceptable carrier.
Example : 7
The stabilized wound healing composition of the present invention comprises of 55% of neutral Succinylated Collagen, 0.15 % of Gentamicin, 45% of Aloevera, 0.4% of Preservative and pharmaceutically acceptable carrier.
Example : 8
The stabilized wound healing composition of the present invention comprises of 45% of neutral Succinylated Collagen, 0.2 % of Gentamicin, 55% of Aloevera, 0.45 % of Preservative and pharmaceutically acceptable carrier.
Example : 9 Clinical studies
The clinical and safety study results of the wound healing composition of the present invention showed that there was no treatment related clinical signs and abnormality observed during the entire period of study. The study did not produce any significant biochemical and histological changes. Hence the composition of the present invention was found to possess significant wound healing activity.
Example 9 a- Burn healing activity of the formulation of the present invention.
In this study burn injury patents were subjected to the formulation of the present invention. Patients were instructed to apply 5gms of the formulation of the present invention on the injured area on daily basis and it was found that the wound was healed completely within 10 days. Figure 1 depicts the burn wound healing process.
Example 9 b- Cut injury healing activity of the formulation of the present invention.
In this cut injury patents were subjected to the formulation of the present invention. Patients were instructed to apply 1gms of the formulation of the present invention on the injured area on daily basis and it was found that the wound was healed completely within 15 days. Figure 2 depicts the cut injury healing process.

Example : 10 Synergism studies
Title : To Study the effect of the formulation of the present invention viz-a-viz its components when individually applied..
Succynilated Collagen/ Aloevera / Gentamicin combination in rat full thickness wound model:
Sonsor : Cologenesis Healthcare P Ltd
Randomisation and Grouping:
Total No.of Animals : 15
Table 1: Randomisation and Grouping:
Groups Treatment
I Control
2 Succ. Collagen gel
3 Gentamicin Cream
4 Aloevera Gel
5 Treatment with our Formulation of the invention.

Study Detail:
All Rats were anaesthetized by injection of Ketamine/ Xylazine . dorsum of the rats was shaved and scrubbed with povidone Iodine solution. Full thickness skin wound of 1 X 1 cms size was created surgically on the dorsum. The wound was infected by MRSA with 40 mic.lit of 2*10^9 CFU/ml and covered with Aluminium foil and then further covered with bandage tape.
The gel/ cream was applied every alternate days. The quantity of gel/ cream applied is each 3 Gms. per application.
The Wound was measured on Day 1, 6,11,16 and 21 Days.
All animals were observed on daily basis for any abnormal clinical signs upto end of the study.
Results:
There was no adverse clinical signs in any animal upto the end of the study. And there was no mortality in any group upto end of the study.
Wound Measurement Detail :
The wound was measured on Day 1, 6,11,16 and 21. The data obtained were tabulated below:
Table 2: Synergism studies
Animal No. Group Day-1 Day-6 Day-11 Day-16 Day-21
L W L W L W L W L W
1 1 14.5 13.2 15.7 11.3 7.8 7.3 2.6 1.6 1.6 1
2 1 12.7 11.8 12.7 12.8 6.9 8.6 4 5.7 2.6 2.2
3 1 12.9 12.4 16.3 11.6 6.7 6.8 2.4 2.2 2 1.8
4 2 12.4 14.5 12.3 14 5.3 4.8 2.1 1.8 0.5 0.7
5 2 15.4 11.3 15.1 12 6.1 4.8 1.9 0.8 0.9 0.5
6 2 12.1 13.3 12.3 13 4.9 4.4 1.6 1.1 0.9 0.4
7 3 14.2 12.4 15.2 13.1 6.6 5.9 2.9 2.7 1 1.1
8 3 12.4 14.4 14.2 15.8 7.4 5.9 2.6 3 1.8 1.9
9 3 15.3 11.3 16.2 13.8 6.3 4.8 3.2 2.4 2.4 1.7
10 4 12.4 14.4 14.2 15.1 7.1 8.3 3.7 4.1 1.4 1.2
11 4 12 13.2 13.2 14 5.7 6.1 3 4 2 1.6
12 4 11.7 12.9 13.5 14.3 5.4 6.6 2.5 3.1 1.1 1.4
13 5 9.95 11 10.5 10.5 1.01 0.52 0.5 0.7 0.5 0.5
14 5 13.2 12.6 9.6 9.5 2.08 2.01 1 0.9 0.4 0.5
15 5 12.6 10.9 14.8 10.7 1.07 0.78 0.7 0.5 0.5 0.5

L= Length in mm W= Width in mm

Conclusion:
Study results clearly indicates that the wound healing is much faster in the new formulation treatment group animals as compared to control group animals.
MRSA infection in all animals belong to all the groups was induced on day 1 and then after the treatment was initiated from Day 3 . All the wounds were covered with gel / cream to cover the entire wound. At each application of gel/ cream, wound healing was observed in each animal by measurement.
Hence it is concluded that the MRSA infected animals belonging to the treatment dose group have shown faster recovery for wound healing as compared to other groups including the control group with MRSA infected animals. (The complete healing was noticed in Day 11 itself, where as the other treatments shows complete healing during 21 days only) This also explains that the formulations of the present invention exhibits significant wound healing activity, compared to the other conventional treatments. This illustrates the synergism of the components present in the composition.

Parameters Neutral Succinylated Collagen Aloevera Gentamicin Formulation of the present invention
Minimum workable dosage 20gms 15gms 8gms 1-5gms
Wound healing activity in percentage
93% 92% 89% 99%
Time required for complete healing 21days 20 days 21 days 15 days
Stability 6 months stored at 40C 1 year 2year 2year

Table 3 Synergism studies

From the above data, it is inferred that for treatment with the composition of the present invention requires only minimum dosage amount when compared to individual constituents. This facilitates reduction in the quantity constituents used. The formulation results in faster and complete healing of wound/ulcer compared to individual constituents and previously reported methods available in public domain. The formulation of the present invention exhibits significant wound healing activity, and this feature enhances the efficacy to treat subjects with wound/ulcer. Further the formulation is highly safe and is free of any side effects compared to other methods. The above discussion illustrates the synergism of the components present in the composition.

From the foregoing it will be appreciated that, although specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without deviating from the spirit and scope of the invention. Accordingly, the invention is not limited except as by the appended claims.

We Claim,
1. A stabilized wound healing composition without the side effect of burning sensation, the claimed composition comprises of therapeutically effective amounts of neutral Collagen, Gentamicin, Aloevera, Preservative and pharmaceutically acceptable carrier wherein the said composition is capable of contacting the wound or to an open ulcer and thereby the collagen breaks down and the Gentamicin is released into the wound or ulcer.

2. The stabilized wound healing composition as claimed in claim 1 wherein the said neutral Collagen, Gentamicin, Aloevera, Preservative and pharmaceutically acceptable carrier being present therein in a proportion producing a synergistic activity with regard to the wound healing activity conferred.

3. A stabilized wound healing formulation without the side effect of burning sensation, the claimed formulation comprises of 40 to 65 % of neutral Collagen, 0.1 to 0.2 % of Gentamicin, 35 to 60 % of Aloevera, 0.3 to 0.55 % of Preservative and pharmaceutically acceptable carrier wherein the said formulation is capable of contacting the wound or to an open ulcer and thereby the collagen breaks down and the Gentamicin is released into the wound or ulcer

4. A stabilized wound healing formulation without the side effect of burning sensation, the claimed formulation comprises of 40% of neutral Collagen, 0.1 % of Gentamicin, 60% of Aloevera, 0.5% of Preservative and pharmaceutically acceptable carrier wherein the said formulation is capable of contacting the wound or to an open ulcer and thereby the collagen breaks down and the Gentamicin is released into the wound or ulcer.

5. The said neutral collagen in any of the preceding claims includes neutral succinylated collagen

6. The said neutral succinylated collagen of claim 5 is prepared by a process comprising
a. Harvesting collagen from –Bovine Tendon/Fish bladder/porcine Tendon
b. cutting and mincing collagen to formed minced collagen
c. subjecting the minced collagen to non-ionic detergent treatment, alkaline treatment followed by enzymatic treatment to obtain normal tissue
d. carrying out succinylation reaction of obtained normal tissue in neutral pH medium to form swell neutral succinylated collagen wherein additional succinyl group is added to prevent subsequent cleavage of lysil peptide bonds with trypsin as a result of succinylation and thereby making the modified protein acts as better substrate for proteases.

7. The wound healing composition/formulation in any of the preceding claims, wherein said pharmaceutically acceptable carrier is an ointment, gel, white petrolatum, light mineral oil, or mixtures thereof.
8. The wound healing composition/formulation in any of the preceding claims wherein the said composition /formulation is capable of evenly spreading over the wound/ulcer for enhanced efficacy.
9. The wound healing composition/formulation in any of the preceding claims wherein the said composition /formulation is stable for atleast 24 months without decomposing into liquid or precipitate.

Dated this 22nd day of June 2015
For COLOGENESIS HEALTHCARE PRIVATE LIMITED
By its Patent Agent
Dr.B.Deepa