CLIAMS:We Claim:
1. A composition for burns and chronic wounds comprising an oil phase, aqueous phase and an active phase;
wherein said oil phase comprising component(s) selected from a group comprising cetomacrogol 1000, cetostearyl alcohol, liquid paraffin, white petroleum jelly, propylene glycol, isopropyl myristate or combinations thereof;
aqueous phase comprising component(s) selected from a group comprising sodium methyl paraben, sodium propyl paraben, EDTA sodium, citric acid or combinations thereof and purified water (Qs);
an active phase comprising a peptide of sequence id no. 1.
2. The composition as claimed in claim 1, wherein said oil phase is present in an amount ranging from 2 to 96% by wt.
3. The composition as claimed in claim 1, wherein said active phase present in an amount ranging from 0.01 to 2.5% by wt.
4. The composition as claimed in claim 1, wherein said aqueous phase is present in an amount ranging from 0.5 to 2% by wt.
5. The composition as claimed in claim 1, wherein said cetomacrogol 1000 is present in an amount ranging from 2 to 10%.
6. The composition as claimed in claim 1, wherein said cetostearyl alcohol is present in an amount ranging from 2 to 10%.
7. The composition as claimed in claim 1, wherein said liquid paraffin is present in an amount ranging from 2 to 7%.
8. The composition as claimed in claim 1, wherein said white petroleum jelly is present in an amount ranging from 15 to 50%.
9. The composition as claimed in claim 1, wherein said propylene glycol is present in an amount ranging from 2 to 9%.
10. The composition as claimed in claim 1, wherein said isopropyl myristate is present in an amount ranging from 2 to 10%.
,TagSPECI:Field of the Invention:
The present application is being filed as patent of addition application on the main patent application 4462/CHENP/2007.
The present invention provides an effective composition for treating burns, diabetic wounds and other type of chronic wounds including treatment of eroded or cracked tissue. The composition shows enhanced therapeutic effect in wound healing and controlling infection rate.
Background of the Invention:
The process of healing of open wounds and burns is a very complex, not fully understood process. Nowadays the most common means of treating open wounds or burns mainly comprise temporary wound closure/covering for protection against e.g. moisture loss and infection etc. Both the nature of the wound and microorganism-specific factors influence the rate of microbial proliferation in and penetration of the wound tissue. The wound as rich in coagulated protein and well hydrated by the trans-eschar movement of fluid and serum, creates an excellent microbial culture medium. Bacterial proliferation in the wound can also be enhanced by factors such as wound maceration, pressure necrosis, and wound desiccation with neo-eschar formation. In addition, secondary impairment of blood flow to the wound could further predispose the patient to invasive infection by curtailing the delivery of oxygen, nutrients, and phagocytic cells to the viable sub-eschar tissue.
The character of the microbial flora of the burn wound changes with time and Gram-negative organisms replace Gram-positive organisms that predominate in the early post wound period by the second week. Without the application of topical antimicrobial agents, the density of bacteria progressively increases, and the microorganisms penetrate the eschar by migration along sweat glands and hair follicles until they reach the eschar/nonviable tissue interface. If the density and invasiveness of the microorganisms exceed the host's defense capacity, proliferating organisms in the sub-eschar space can invade the underlining viable tissue, leading to invasive burn wound infection and even systemic spread to remote tissues and organs. Bacterial invasion is uncommon unless the number of microorganisms exceeds 105/grams of biopsy tissue. Over the past 20 years, significant changes in the microbial ecology of the burn has been observed. The recovery of Pseudomonas and other gram-negative bacteria, which were once the most common organisms causing burn wound infection, has markedly declined with improvements in the isolation of patients. Patients who have received broad-spectrum antibiotics for perioperative coverage or treatment of septic complications and whose wounds remain open for many days owing to the extent of the burn are at increased risk of burn wound colonization and infection by yeasts, fungi, and multiple antibiotic-resistant bacteria. The true fungi have replaced bacteria as the most common microbes causing burn wound infection in recent years. Pseudomonas aeruginosa, Staphylococcus aureus and Aspergillus species are the commonest organisms associated with burn wound infection.
The current treatment of the burned patient involves fluid resuscitation, hemodynamic stabilization and the application of topical antimicrobials prior to wound excision and grafting. Bacteria flourish in dead tissue and intravenous antibiotics cannot penetrate this tissue because of its impaired blood supply. This necessitates the use of topical antimicrobials that are meant to curtail the systemic invasion of bacteria. Unfortunately, the commonly employed topical antimicrobials are often not enough to prevent the fatal consequences of burn wound sepsis. Synthetic antimicrobial peptides are specific proteins designed to encompass a greater spectrum of activity against bacteria (including multi-drug resistant organisms) and fungus than traditional topical agents. It is clear that the topical agents are crucial in the ultimate eradication of the burn and infected wound pathogens since it is extremely difficult to administer the intravenous antibiotics to non-perfused tissue such as burned skin. The poorly vascularized, wounded skin is, therefore, the portal of entry and the ongoing nidus of infection for burn victims. The ideal topical agent should be highly active against common and multi-resistant pathogens, such as methicillin resistant Staphylococcus aureus, vancomycin resistant Enterococcus faecium/faecalis, and extended spectrum ß-lactamase producing Gram- negative organisms, while having a neutral or even beneficial effect on the wound healing process.
The currently existing treatment options include compositions based on active agents like Mafenide acetate, 1% SSD and silver nitrate. These three are the most commonly employed topical antimicrobial agents for burn wound care. However, each of these agent has specific limitations and advantages like 1% SSD burn cream is most effective when applied to burns soon after injury to minimize bacterial proliferation on the wound's surface but it has limited solubility in water and, therefore, limited ability to penetrate into the eschar. Also, with continual use, resistance to the sulfonamide component of 1% SSD is common, particularly in certain strains of Pseudomonas and many Enterobacter species.
While, the other agent Mafenide acetate diffuses freely into the eschar owing to its high degree of water solubility. It is the preferred agent if the patient has heavily contaminated burn wounds or has had burn wound care delayed by several days. Hypersensitivity reactions occur in 7% of patients. However, the use of this agent causes inhibition of carbonic anhydrase, and promotes diuresis of bicarbonate as observed after its use. The resultant metabolic acidosis could accentuate post-burn hyperventilation, and significant acidemia could develop if compensatory hyperventilation is impaired.
Similarly, use of 0.5% silver nitrate solution has a broad spectrum of antibacterial activity imparted by the silver ion. However, this agent does not penetrate the eschar because the silver ions are rapidly precipitated on contact with any protein or cationic material.
There are various patent applications relating to the composition for the treatment of burns which are based on the herbal technology. WO 2012151438 provides an antimicrobial silver hydrogel composition useful for the treatment of burns and wounds.
The composition comprises an anhydrous, hydrous stabilized, silver salt and aloe vera gel in an aqueous vehicle.
Similarly, WO 2003015808 provides a medicinal composition for treating pressure sore ulcerations and other skin conditions. The composition is based on herbs comprising Prunus armeniaca, Moras albae, Ledebouriella seseloides, Artemisia argyi, Schizoneptatenuifolia, Cnidium monnieri, Alum and Glycyrrhiza uralensis.
However, the herbal based compositions have their limitation in term of efficacy in wound treatment and sometimes may fail to meet the desired therapeutic tendency.
Despite of various ongoing developments in the domain for finding an appropriate strategy treatment for burns/wound treatment, there is still a need for a highly efficient composition which could show promising effects in healing of chronic wounds/diabetic wounds/cracked or eroded tissues etc. along with showing improved effect in terms of reduced itching, pain, distress upon change of dressing on to wound area.
Object(s) of the Invention:
A primary object of the present invention is to overcome the drawbacks of the prior art.
Yet another objective of the present invention is to provide a highly efficient composition for the treatment of burns/wounds/diabetic ulcer.
Yet another objective of the present invention is to provide an efficient composition with improved wound healing properties.
Yet another objective of the present invention is to provide a highly efficient composition with improved infection clearance rate.
Yet another objective of the present invention is to provide a composition in the form of a cream with good textural effects.
Yet another objective of the present invention is to provide a composition with the ability to produce minimal scar, itching and breaking in/around the skin of wound area.

Summary of the Invention:

In one aspect of the Invention, there is provided a composition for burns and chronic wounds comprising an oil phase, aqueous phase and an active phase;

wherein said oil phase comprising component(s) selected from a group comprising cetomacrogol 1000, cetostearyl alcohol, liquid paraffin, white petroleum jelly, propylene glycol, isopropyl myristate or combinations thereof;
aqueous phase comprising component(s) selected from a group comprising sodium methyl paraben, sodium propyl paraben, EDTA sodium, citric acid or combinations thereof and water;
an active phase comprising a peptide of sequence id no. 1 .
Brief description of drawing(s):
Figure 1(a) and (b): illustrates the photographic comparison of treatments performed with 1%SSD and the composition of present invention
Figure 2: illustrates the efficacy of composition for the treatment of diabetic wound
Figure 3: illustrates the efficacy of composition for the treatment of chronic wounds
Detailed description of the Invention:
The present application is filed as “patent of addition” over the main invention protected in the patent application no. 4462/Chenp/2007.
The present invention in accordance with the above stated object(s) provides an improved composition (herein after referred as XYLENTRA throughout the specification), specifically useful for the treatment of burns and chronic wounds such as those found in diabetic wounds and eroded or cracked tissue particularly on the extremities.
The composition shows promising results with significantly accelerated wound healing effect.
The present application comprises improvement or modification of the invention claimed in the specification of the main patent application bearing no. 4462/Chenp/2007, in the following manner:
• The composition shows improved wound healing and controlled infection rate
• The composition is efficacious in controlling non-viable tissue and granulation in the process of wound healing
• The composition is effective in controlling edema, erythema and purulence
• The composition shows reduced scar, itching and breaking in/around the skin of wound area and promotes the scar maturation and normal skin formation
• The clinical trials show that the patients treated with the composition of present application returned to their normal skin color without any scar or contracture formation
• Significantly less hyper pigmentation and hypo pigmentation was observed in the patient treated with the composition of present invention
• Clinical trials show very less itching in the wound area of the patient treated with the composition
The invention is further described in detail as below:
The composition comprises an oil phase, aqueous phase and an active phase. The oil phase comprises component(s) selected from a group comprising cetomacrogol 1000, cetostearyl alcohol, liquid paraffin, white petroleum jelly, propylene glycol, isopropyl myristate or any combinations thereof.
The aqueous phase comprises component(s) selected from a group comprising sodium methyl paraben, sodium propyl paraben, EDTA sodium, citric acid or combinations thereof and purified water.
While, the active phase comprises a peptide with sequence id no.1 having following amino acid sequence:
FAKKFAKKFAKFAKKFAKFAFAF
The surprising effect achieved by the present composition is due to the specific components i.e. components of oil phase, aqueous phase and the active phase, which are customized to be present in a particular amount of range, thus imparting an enhanced healing effect. The composition also have desirable sensory characteristics like textural properties, thus allowing the easier and smoother application of the composition.
The oil phase is present in an amount ranging from 2 to 96% by wt. while the aqueous phase is present in an amount ranging from 0.5 to 2% by wt, purified water (QS) while the active phase is present in an amount ranging from 0.01 to 2.5% by wt.
The composition is preferable in the form of a cream. However, the composition could be formulated in any other appropriate topical forms as well.
The components of oil phase are present in following particular amount range:
a. Cetomacrogol 1000, acting as Solubilizer, Emulsifying Agent, Non Ionic surfactant as it helps in lowering the surface tension between two liquids) is present in an amount ranging from 2 to 10%.
b. Cetostearyl Alcohol acting as emulsion stabilizer and viscosity increasing agent is present in an amount ranging from 2 to 10%.
c. Liquid paraffin acting as moisturizer is present in an amount ranging from 2 to 7%.
d. White petroleum jelly, acting as Ointment base and useful for soothing application is present in an amount ranging from 15 to 50%.
e. Propylene glycol, acting as humectants to keep the skin moist is present in an amount ranging from 2 to 9%.
f. Isopropyl myristate acting as emollient for softening and soothing effect to skin is present in an amount ranging from 2 to 10%.
The aqueous phase comprises Sodium Methyl Paraben, Sodium Propyl Paraben, EDTA sodium and Citric acid which are present in an amount ranging from 0.5 to 2% by wt. The aqueous phase further comprises purified water(Qs).
Sodium Methyl Paraben is present as a preservative. In the composition, sodium propyl paraben with sodium methyl paraben shows synergistic effect.
While active phase comprises a peptide of sequence id 1 i.e. Genopep 1. The peptide showed significant antibacterial activity on test organisms Staphylococcus aureus MTCC 96 and Pseudomonas aeroginosa MTCC 741. The test compound showed 100% killing of Staphylococcus aureus on exposure to 1µM (4.3 µg/ml) and 5 µM (21.5 µg/ml) concentrations for 1 hr at pH 7.2, and at pH 8.4 an exposure of 4 hrs was required to get 100% killing. Whereas, 100% killing of Pseudomonas aeruginosa was observed on exposure to the test compound for 1 hr at pH 8.4 and an exposure of 4 hrs was required for 100% killing at pH 7.2.
The microbiological studies with “Xylentra” in-vivo using a rat burn wound model were also conducted. The observations on the bacterial growth in eschar and sub-eschar muscles on post burn day one, two or three in peptide treated and control treated groups were made. A substantial decrease in the microbial population level was observed in animals treated with peptide.
Sub Acute Toxicity studies of (Xylentra) in rats and rabbits demonstrated that it was safe. No abnormalities in physical, physiological, biochemical and histo-pathological parameters were observed on application of the peptide. No mortality was seen in animals from any group. There is evidence (dermal histopathology findings) to show that Xylentra® has stimulatory action on tissue growth (increased collagen content in granulation tissue and re-epithelialization) thus promoting improved wound healing.
The results of a Phase-I clinical trial (n = 24) on healthy human patients revealed that Xylentra cream when administered topically twice a day was safe. There was an absence of any symptoms or signs of toxicity at the evaluation site and considered a measure of primary efficacy / tolerability was established in this study. The composition was safe and adverse events were found to be minimal in the Phase-I Study. Mild redness and itching was seen in 2/24 volunteers. Treatment Groups (2% and 4%) were similar in efficacy/ tolerability and safety parameters studied.
While, phase II study (n = 120) with the Xylentra, NO Treatment Groups (0.02% & 0.05%) reveals that the incidence rate of wound healing was better in terms of time to complete wound healing and achievement of a superior overall wound evaluation score over the Placebo Group. Scar formation was also significantly lower compared to that in Placebo Group, indicating that treatment with Xylentra enabled more complete healing with less morbidity. It is safe and highly effective in promoting burn wound healing for patients with partial thickness burns that are less than or equal to 20% without any side effects. None of the Xylentra treated volunteers developed any adverse reaction. Pharmacokinetic data suggested the absence of systemic absorption.
Yet another study was performed for comparing safety and efficacy of 1% SSD (composition existing in the market) with Xylentra in patients with partial thickness burns in India.
One hundred and sixty patients completed the study. The objective was to compare the safety and efficacy of Xylentra with Silver Sulfadiazine in patients with partial thickness burns. The patients were topically administered with either Xylentra or Silver Sulfadiazine cream based on allocation concealment once daily for 4 weeks or until complete wound healing whichever is earlier.

The study met its primary endpoints. The proportion of patients with complete wound closure was statistically significant between the Xylentra and Silver Sulfadiazine group. The patients with complete wound closure were 75 (94%) (N=80) and 65 (81%) (N=80) in the Xylentra and Silver Sulfadiazine groups, respectively in the four week time period. Moreover, Xylentra demonstrated accelerated healing rates when compared to Silver Sulfadiazine. The time taken for complete wound closure was found to be less in Xylentra group compared to Silver Sulfadiazine group as more number of patients had complete wound closure at various time points in the Xylentra group when compared to the Silver Sulfadiazine group. Cumulative wound healing was found to be better in Xylentra group when compared to Silver Sulfadiazine group. Cumulative wound healing was 85.42% Xylentra group and 78.71% in Silver Sulfadiazine.

Burn wounds are a richer source of infection than surgical wounds, primarily because of the larger area involved and longer duration of patient stay in the hospital. In this study, clinical and statistically significant differences were found in the proportion of patients with controlled secondary infections between the groups. Xylentra showed increased efficacy in controlling the infection rate as evident from 41.7% of patients with controlled secondary infection in the Xylentra composition group when compared to 25% of patients with controlled secondary infection in Silver Sulfadiazine group. Additionally, antibiotic usage was lower by patients in the Xylentra group when compared to the Silver Sulfadiazine group.

Most commonly encountered and clinically significant impediments to wound healing are the presence of non-viable tissue and granulation. Xylentra proved to be efficacious in controlling both non-viable tissue and granulation throughout the study period. Both the evaluation of mean score of non-viable tissue and patient wise evaluation confirmed that Xylentra remarkably reduced non-viable tissue and granulation in comparison to Silver Sulfadiazine. On study termination day, no non-viable tissue was found in 78 (97.50%) and 71 patients (88.75%) in Xylentra and Silver Sulfadiazine groups, respectively. It was clinically observed that non-viable tissue became necrotic tissue in the Silver Sulfadiazine treated patients. However non-viable tissue appeared during the initial visits in Xylentra treated patients but no necrotic tissue was observed during the study period. Similarly, at the end of study termination day, the number of patients observed with no granulation in Xylentra group was 75 (93.75%) and 63 (78.75%) in the Silver Sulfadiazine group.

Xylentra also proved to be potent in wound evaluation based on clinical parameters. It was effective in controlling edema, erythema and purulence as evident from the results of mean score analysis and patient wise analysis. Patients with no edema, no erythema and no purulence were found to be less in number in Xylentra group when compared to Silver Sulfadiazine group. On study termination day, no erythema was found in 78 (97.50%) and 74 patients (92.50%) in Xylentra and Silver Sulfadiazine groups respectively. No edema was found in 79 (98.75%) and 74 (92.50%) in Xylentra and Silver Sulfadiazine groups, respectively. No purulence was found in 78 (97.50%) and 74 (92.50%) patients in Xylentra and Silver Sulfadiazine groups, respectively. In addition, the mean scores of erythema, edema and purulence were found to be considerably lower in Xylentra group in comparison to Silver Sulfadiazine group.

In addition to these primary efficacy and secondary efficacy endpoints, other variables that demonstrate the potency of the drug such as drug’s ability to reduce scar (pigmentation and thickness), itching and breaking were evaluated. Scar maturation and normal skin formation always will be depending on the quality of wound healing. Patients of post-burn contractures, scars defects and disfigurements constitute almost half of the workload of many general plastic surgeons; especially this will directly increase the cost to patient for surgical procedures. Hence choosing the quality healing medicine is very important step in burn wound management. The clinical observations showed that most of the Xylentra treated patients’ wound area returned to their normal skin color without any scar or contracture formation. However, most of the patients treated with Silver Sulfadiazine had dark pigmentation with hypertrophic scars with contractures. Hyper pigmentation and hypo pigmentation was significantly less in Xylentra group (13% and 16%) when compared to Silver Sulfadiazine group (25% and 21%). The area with paper thin and hypertrophic scar was more than twice in Silver Sulfadiazine group (6% and 25%) when compared to Xylentra group (2% and 12%). Itching was found significantly greater in treated patients (51%) in Silver Sulfadiazine group in comparison to Xylentra group (14%). In addition, 18% of treated patients in Silver Sulfadiazine group had breaking of skin in comparison to 3% of treated patients in Xylentra. These results demonstrate the fact that Xylentra has the ability to heal the wound effectively and controlling the development of scars, pigmentation, itching and breaking.

The treatment emergent adverse events (TEAEs) observed were mostly skin related disorders. All the adverse events were found to be mild in nature. No serious adverse events and no deaths were reported in the study. The adverse events frequently experienced by patients in the study were swelling, irritation, itching, redness and pain. All these adverse events were observed in high percentage of patients and were present for longer duration in Silver Sulfadiazine group when compared to Xylentra group.

A summary of the statistical measurements of the test variables is listed in Table 1 while the photographic comparison of both treatments can be found in Figure 1A and B.

Figure 1(a) and (b) shows hypopigmentation and scarring effect present in the patients treated with 1% SSD while the patient treated with the composition of present invention does not show any scarring effect and the pigmentation also returns to the normal level.

Figure 2 shows the efficacy achieved by the present composition in the treatment of Diabetic wound. The patients were treated with the composition cream once per day for the indicated times listed in Figure. The efficacy can clearly be seen as significant healing has occurred over the test period. Diabetic wounds are notoriously difficult to treat. Oftentimes, the only recourse is amputation.

Figure 3 shows the efficacy achieved by the present composition in the treatment of Chronic Wounds. The test was conducted for the treatment of Cracked Feet Experiments where patients were treated with the composition cream once per day for the indicated times listed on Figure 3. As evident, the composition shows good results.

Primary Variables p-value Significant
Variable 1
% Patients achieving complete wound closure/healing 0.0168 YES
Variable 2
% Patients with controlled secondary infections
(on day 20) 0.0198 YES
Secondary Variables
Variable 1
% of wound covered with non-viable tissue 0.0036 YES
Variable 2
Degree of granulation 0.0001 YES
Variable 3

Summary of Statistical Measurements of Test Variables
Table 1
The synergistic effect of the composition is further described with the help of following negative experiments performed to observe the therapeutic efficacy.
Formulation I with following components is prepared:
Components:
• Active phase: Xylentra Cream 0.05% i.e. peptide of sequence Id 1;
• Size: 500gr
• B.No:ISCT.001 (Naik)
S.No INGREDIENTS
OIL PHASE
1 White Petroleum jelly
2 Steryl Alcohol
3 Liquid Paraffin
4 Stearic Acid
5 Glycerin
6 Isipropyl Myristate
AQUEOUSE PHASE
1 Methyl Paraben Sodium
2 Propyl Paraben Sodium
3 EDTA
4 Purified Water
ACTIVE PHASE
1 *Genopep 1
2 Purified Water
Observation: Texture of the cream was not good & there is no smoothness

Another formulation II with following components was prepared

• Active phase: Xylentra Cream 0.05% i.e. peptide of sequence Id 1;
• Batch Size: 1.0kg.
• Mfg Dt:Sep.2014
• Exp Dt: Aug.2016
PHASE S.No INGREDIENTS
OIL PHASE 1 Cetomacrogol 1000
2 Steryl Alcohol
3 Liquid Paraffin
4 White Petroleum jelly
5 Propylene Glycol
AQUEOUSE PHASE 6 Sodium Methyl Paraden
7 Sodium Propyl Paraben
8 Sodium Phosphate
9 Sodium Acid Phospate
10 EDTA Disodium
11 Purified Water
12 Caffeine
13 Citric Acid

Observation: The cream was too waxy while rubbing
Yet another formulation III with following components was prepared:
Composition of Cream
S. No. Components Amount(%)
1 Cetomacrogol-1000 2-5 %
2 Stearyl Alcohol 8-15 %
3 Liquid Paraffin 3- 6%
4 White Petrolium Jelly 15-20 %
5 Propylene Glycol 6-10 %

6 Peptide of sequence Id 1 0.05- 2.0 %
7 Sodium Methyl Paraben 0.1-0.2 %
8 Sodium Propyl Paraben 0.05- 0.1 %
9 Sodium Phosphate 0.1-0.3 %
10 Sodium Acid Phosphate 0.01-0.03 %
11 EDTA Disodium 0.01-0.03 %
12 Citric Acid 0.3 – 0.8 %
13 Purified Water 50-65 %
Observation:
As evident, that enhanced healing effect is achieved only when the oil phase, aqueous phase and the active phase is present in the prescribed amount of the present composition. The above stated formulation I, II and III shows about the adverse effect achieved when the components are not present as per the composition of present invention.
The invention will be described with the help of non-limiting examples:
Example 1:
An initial double blind randomized, phase II/III clinical trial for the evaluation of a cream based peptide preparation in the topical treatment of burn wounds was conducted at Gandhi Medical College Hospital in Hyderabad, India in 2008. It was observed that the mean duration to reach the total wound closure for peptide treated patients was found to be 8.4 days while the placebo group was 11.1 days. This was statistically significant at the (P<0.001). A second study consisting of 120 burn patients, begun in late 2008 was carried out at two hospitals in Hyderabad, India. In these studies the patients received no additional antibiotic therapy and none of the wounds were debrided. The final report indicated that there were no infections and significant accelerated healing with minimal scarring and the patients had significantly fewer hospital visits.
Example 2:

Example 3: