SPRAY COMPOSITIONS OF CHITOSAN FOR WOUND HEALING

Abstract

The present invention relates to topical spray compositions comprising chitosan and the process for preparation thereof. The topical spray compositions comprising chitosan are used for the treating burn wounds, wounds resulting from chemical burns, wounds from physical trauma, neuropathic ulcers, pressure sores, diabetic ulcers. Further the spray compositions composition is used for treating gynaecological diseases, urogenital infections (vaginal bacteriosis, urinary tract infection and vaginitis) with good effect, safe, natural, and good biocompatibility by restoring and/or maintaining the pH of vagina. More specifically the present invention relates to topical spray compositions comprising chitosan and carbonic acid and the process for preparation thereof.

Claims

1. A topical spray composition comprising chitosan and carbonic acid.

2. The topical spray composition according to claim 1, wherein the composition comprises about 0.01% w/v to about 1.0% w/v of chitosan.

3. The topical spray composition according to claim 1, wherein the composition comprises about 0.1% w/v to about 1.0% w/v of carbonic acid.

4. The topical spray composition according to claim 1, wherein the composition further comprises lactic acid and water.

5. The topical spray composition according to claim 1, wherein the spray composition is stored in a pressurized vessel.

6. The topical spray composition according to claim 5, wherein pressurized vessel is a Bag-On-Valve (BOV) aerosol dispensing system.

7. A topical spray composition comprising chitosan, carbonic acid, lactic acid and water.

8. The topical spray composition according to claim 7, wherein the composition comprises of about 0.01% w/v to about 1.0% w/v of chitosan, about 0.1% w/v to about 1.0% w/v of carbonic acid, 0.01% w/v to 2.0% w/v of lactic acid and water.

9. The topical spray composition according to claim 8, wherein the spray composition is stored in a pressurized vessel.

10. The topical spray composition according to claim 9, wherein pressurized vessel is a Bag-On-Valve (BOV) aerosol dispensing system.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0049] FIG. 1 discloses the rate of wound contraction (healing) calculated of the present inventive composition [chitosan in carbonic acid and water (carbonated water) as disclosed in Example-1] in comparison to chitosan solution (without carbonation), carbonic acid in water (carbonated water), reference and normal control (untreated).

[0050] FIG. 2 discloses the average wound breaking strength of Example-1 in comparison to chitosan solution (without carbonation), carbonated water, reference and normal control (untreated).

DETAILED DESCRIPTION OF THE INVENTION

[0051] The present invention relates to the topical spray composition comprising chitosan and carbonic acid.

[0052] In a specific embodiment, the present invention contains chitosan. The molecular weight of the chitosan used in the present invention is in range of about 50,000 to about 100,000. In the topical spray composition, chitosan is present in the range of about 0.01% w/v to about 1.0% w/v. More preferably chitosan used in the topical spray composition is about 0.01% w/v, 0.015% w/v, 0.02% w/v, 0.025% w/v, 0.03% w/v, 0.04% w/v, 0.045% w/v, 0.05% w/v, 0.055% w/v, 0.06% w/v, 0.065% w/v, 0.07% w/v, 0.075% w/v, 0.08% w/v, 0.085% w/v, 0.09% w/v, 0.095% w/v, 0.1% w/v, 0.15% w/v, 0.20% w/v, 0.25% w/v, 0.30% w/v, 0.40% w/v, 0.50% w/v, 0.60% w/v, 0.70% w/v, 0.75% w/v, 0.80% w/v, 0.90% w/v and 1.0% w/v. Most preferably chitosan used in the topical spray composition is of about 0.05% w/v to about 0.1% w/v and even most preferably chitosan used in the topical spray composition is of about 0.05% w/v, 0.055% w/v, 0.06% w/v, 0.065% w/v, 0.07% w/v, 0.075% w/v, 0.08% w/v, 0.085% w/v, 0.09% w/v, 0.095% w/v, 0.1% w/v.

[0053] In embodiments of the present invention, the inventive topical spray composition comprises carbonic acid. Carbonic acid used in the present invention is in range of about 0.1% w/v to about 1.0% w/v. More preferably carbonic acid used in the spray composition is about 0.2% w/v to about 0.4% w/v of carbonic acid and most preferably carbonic acid used in the spray composition is of about 0.2% w/v, 0.25% w/v, 0.3% w/v, 0.35% w/v and 0.4% w/v.

[0054] The present invention further provides the topical spray composition comprising chitosan, carbonic acid, lactic acid and water.

[0055] In embodiments of the present invention, the inventive spray composition comprises lactic acid. Lactic acid is used to dissolve the chitosan. Lactic acid used in the present invention is in range of about 0.01% w/v to about 2.0% w/v. More preferably lactic acid used in the topical spray composition is about 0.01% w/v, 0.02% w/v, 0.03% w/v, 0.04% w/v, 0.05% w/v, 0.055% w/v, 0.06% w/v, 0.065% w/v, 0.07% w/v, 0.075% w/v, 0.08% w/v, 0.085% w/v, 0.09% w/v, 0.095% w/v, 0.1% w/v, 0.15% w/v, 0.20% w/v, 0.25% w/v, 0.30% w/v, 0.40% w/v, 0.50% w/v, 0.60% w/v, 0.70% w/v, 0.75% w/v, 0.80% w/v, 0.90% w/v and 1.0% w/v, 1.05% w/v, 1.1% w/v, 1.15% w/v, 1.2% w/v, 1.25% w/v, 1.3% w/v, 1.35% w/v, 1.4% w/v, 1.45% w/v, 1.5% w/v, 1.55% w/v, 1.6% w/v, 1.65% w/v, 1.7% w/v, 1.75% w/v, 1.8% w/v, 1.85% w/v, 1.9% w/v, 1.95% w/v and 2.0% w/v.

[0056] In embodiments of the present invention, the inventive spray composition comprises water. Water used in the present composition quantity sufficient to 100% w/v of composition.

[0057] In embodiments of the present invention, the present invention, the topical spray composition comprises chitosan, carbonic acid, lactic acid and water, wherein the topical spray composition has a pH of about 2.0 to about 4.5.

[0058] In further embodiments of the invention, the present invention provides the topical spray composition comprising additional excipients selected from glycerin. Glycerin is preferably used in the range from about 0.1% w/v to 1% w/v of the composition and more preferably 0.2% w/v, 0.3% w/v, 0.4% w/v, 0.5% w/v, 0.6% w/v, 0.7% w/v and 0.8% w/v.

[0059] In embodiments of the present invention, the present invention provides the topical spray composition comprising chitosan and carbonic acid stored in a pressurized vessel.

[0060] In further embodiments of the present invention, the present invention provides the topical spray composition comprising chitosan, carbonic acid, lactic acid and water stored in a pressurized vessel.

[0061] In embodiments of the invention, carbonic acid gas continuously introduced into water, carbonic acid gas is dissolved in water to form carbonic acid solution. The topical spray composition according to the present invention is stored in a pressure vessel. Higher pressures may facilitate the dissolved carbonic acid in solution to form a stable solution of chitosan. The topical spray composition of the present invention was charged in vials of high pressure for portable use. By shaking the pressurized container, the solution of carbonic acid gas released, after the discharge nozzle, spray composition quickly escape, which forms a solid film or film on the wound surface for the treating burn wounds, wounds resulting from chemical burns, wounds from physical trauma, neuropathic ulcers, pressure sores and diabetic ulcers or surface of skin for the treatment of urogenital infections (vaginal bacteriosis, urinary tract infection and vaginitis). In embodiments of the invention the pressurized vessel used is high-pressure resistant container bottle with a nozzle in a canning mode (where the composition of the present invention is mixed with propellant like nitrogen and filled into container and sealed) or high-pressure resistant Bag-On-Valve (BOV) aerosol dispensing system. Most preferably the pressurized vessel used in the present invention is Bag-On-Valve (BOV) aerosol dispensing system.

[0062] In the embodiments of the present invention, the composition comprising chitosan, carbonic acid, lactic acid and water is mixed with propellant like nitrogen and/or compressed air and filled into the high-pressure resistant container bottle with a nozzle in canning mode.

[0063] In another embodiment of the present invention, the composition comprising chitosan, carbonic acid, lactic acid and water if filled into the bag, the bag is fitted into the pressurized container containing propellant like nitrogen and/or compressed air and sealed to fitted with actuator to form the bag-on-valve (BOV) aerosol dispensing system.

[0064] In embodiments of the invention, the present invention provides the topical spray composition consisting essentially about 0.01% w/v to about 1.0% w/v of chitosan and about 0.1% w/v to about 1.0% w/v of carbonic acid.

[0065] In another embodiment of the invention, the present invention provides the topical spray composition consisting essentially about 0.01% w/v to about 1.0% w/v of chitosan and about 0.1% w/v to about 1.0% w/v of carbonic acid, wherein spray composition is stored in a pressurized vessel.

[0066] In a further embodiment of the invention, the present invention provides the topical spray composition consisting essentially about 0.01% w/v to about 1.0% w/v of chitosan and about 0.1% w/v to about 1.0% w/v of carbonic acid, wherein spray composition is stored in high-pressure resistant container bottle with a nozzle in a canning mode.

[0067] In a still further embodiment of the invention, the present invention provides the topical spray composition consisting essentially about 0.01% w/v to about 1.0% w/v of chitosan and about 0.1% w/v to about 1.0% w/v of carbonic acid, wherein spray composition is stored in high-pressure resistant Bag-On-Valve (BOV) aerosol dispensing system.

[0068] In a specific embodiment of the invention, the present invention provides the topical spray composition consisting essentially about 0.05% w/v to about 0.1% w/v of chitosan and about 0.2% w/v to about 0.4% w/v of carbonic acid.

[0069] In one more embodiment of the invention, the present invention provides the topical spray composition consisting essentially about 0.05% w/v to about 0.1% w/v of chitosan and about 0.2% w/v to about 0.4% w/v of carbonic acid, wherein the spray composition is stored in a pressurized vessel.

[0070] In another embodiment of the invention, the present invention provides the topical spray composition consisting essentially about 0.05% w/v to about 0.1% w/v of chitosan and about 0.2% w/v to about 0.4% w/v of carbonic acid, wherein the spray composition is stored in high-pressure resistant container bottle with a nozzle in a canning mode.

[0071] In a further embodiment of the invention, the present invention provides the topical spray composition consisting essentially about 0.05% w/v to about 0.1% w/v of chitosan and about 0.2% w/v to about 0.4% w/v of carbonic acid, wherein the spray composition is stored in high-pressure resistant Bag-On-Valve (BOV) aerosol dispensing system.

[0072] In embodiments of the invention, the present invention provides the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 2.0% w/v of lactic acid, about 0.1% w/v to about 1.0% w/v of carbonic acid and water.

[0073] In a further embodiment of the invention, the present invention provides the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 2.0% w/v of lactic acid, about 0.1% w/v to about 1.0% w/v of carbonic acid and water, wherein the spray composition is stored in a pressurized vessel.

[0074] In another embodiment of the invention, the present invention provides the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 2.0% w/v of lactic acid, about 0.1% w/v to about 1.0% w/v of carbonic acid and water, wherein spray composition is stored in high-pressure resistant container bottle with a nozzle in a canning mode.

[0075] In a still further embodiment of the invention, the present invention provides the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 2.0% w/v of lactic acid, about 0.1% w/v to about 1.0% w/v of carbonic acid and water, wherein spray composition is stored in in high-pressure resistant Bag-On-Valve (BOV) aerosol dispensing system.

[0076] In a further embodiment of the invention, the present invention provides the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 2.0% w/v of lactic acid, about 0.1% w/v to about 1.0% w/v of carbonic acid and water quantity sufficient to 100% w/v of composition.

[0077] In another embodiment of the invention, the present invention provides the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 0.2% w/v of lactic acid, about 0.1% w/v to about 1.0% w/v of carbonic acid and water, wherein the spray composition is stored in a pressurized vessel.

[0078] In a further embodiment of the invention, the present invention provides the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 0.2% w/v of lactic acid, about 0.1% w/v to about 1.0% w/v of carbonic acid and water, wherein the spray composition is stored in high-pressure resistant container bottle with a nozzle in a canning mode.

[0079] In a still further embodiment of the invention, the present invention provides to the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 0.2% w/v of lactic acid, about 0.1% w/v to about 1% w/v of carbonic acid and water, wherein spray composition is stored in in high-pressure resistant Bag-On-Valve (BOV) aerosol dispensing system.

[0080] In another of the invention, the present invention provides the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 2.0% w/v of lactic acid, about 0.1% w/v to about 1.0% w/v of carbonic acid and water quantity sufficient to 100% w/v of composition, wherein the spray composition has a pH of about 2.0 to about 4.5.

[0081] In another embodiment of the invention, the present invention provides the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 0.2% w/v of lactic acid, about 0.1% w/v to about 1.0% w/v of carbonic acid and water, wherein the spray composition has a pH of about 2.0 to about 4.5 and wherein the spray composition is stored in a pressurized vessel.

[0082] In a further embodiment of the invention, the present invention provides the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 0.2% w/v of lactic acid, about 0.1% w/v to about 1.0% w/v of carbonic acid and water, wherein the spray composition has a pH of about 2.0 to about 4.5 and wherein the spray composition is stored in high-pressure resistant container bottle with a nozzle in a canning mode.

[0083] In a still further embodiment of the invention, the present invention provides the topical spray composition consisting about 0.01% w/v to about 1.0% w/v of chitosan, about 0.01% w/v to 0.2% w/v of lactic acid, about 0.1% w/v to about 1% w/v of carbonic acid and water, wherein the spray composition has a pH of about 2.0 to about 4.5 and wherein spray composition is stored in in high-pressure resistant Bag-On-Valve (BOV) aerosol dispensing system.

[0084] The following example is provided to illustrate the present invention. It is understood, however, that the invention is not limited to the specific conditions or details described in the example below. The example should not be construed as limiting the invention as the examples merely provide specific methodology useful in the understanding and practice of the invention and its various aspects. While certain preferred and alternative embodiments of the invention have been set forth for purposes of disclosing the invention, modification to the disclosed embodiments can occur to those who are skilled in the art.

Examples 1 to 6

[0085] Spray Composition:

TABLE-US-00001 Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Ingredient (w/v) (w/v) (w/v) (w/v) (w/v) (w/v) Chitosan 0.05% 0.075% 0.1% 0.05% 0.05% 0.05% Lactic acid 0.04% 0.07% 0.14% 0.04% 0.04% 0.04% Carbonic acid 0.4% 0.4% 0.4%  0.4%  0.4%  0.4% Glycerol — — —   1%   3%   5% Water Q.S to Q.S to Q.S to Q.S to Q.S to Q.S to 100% 100% 100% 100% 100% 100% pH 3.5 3.5 3.5 3.5 3.5 3.5

[0086] Process for Preparation of Examples 1-3 [0087] a. Collect 70% batch size of water for injection into suitable manufacturing vessel. [0088] b. Add and disperse the chitosan in step a. [0089] c. Add required quantity of 1 Molar lactic acid slowly till the chitosan get completely solubilized. [0090] d. Makeup the volume to 100% batch size using water for injection. [0091] e. Filter the solution through 0.2μ and cool to below 5° C. [0092] f. Carbonate the solution and immediately fill into the container and closed. [0093] Note: In case of non-aseptic filling, the product shall be sterilized by gamma irradiation or other suitable method of sterilization.

[0094] Process for Preparation of Examples 4-6 [0095] a. Collect 70% batch size of water for injection into suitable manufacturing vessel. [0096] b. Add and disperse the chitosan in step-a. [0097] c. Add required quantity of 1 Molar lactic acid slowly till the chitosan get completely solubilized. [0098] d. Add dispensed quantity of glycerol to step c. [0099] e. Makeup the volume to 100% batch size using water for injection. [0100] f. Filter the solution through 0.2μ and cool to below 5° C. [0101] g. Carbonate the solution and immediately fill into the container and closed.

[0102] Note: In case of non-aseptic filling, the product shall be sterilized by gamma irradiation or other suitable method of sterilization.

Example-7: Procedure for Excision Wound Healing Study

[0103] Rats were anesthetized with ketamine (30 mg/kg, ip) and an area of about ≈500 mm.sup.2 was marked on the back of the rat by a standard ring. Full thickness of the marked skin was then cut carefully. Treatment was started from 1.sup.st day and continued till day 22. Wounds were traced on OHP sheet and calculated by superimposing on graph paper on the day of wounding and subsequently at a gap period of 4 days till 12.sup.th day, then on the alternate days until healing was completed.

[0104] The changes in wound area was measured regularly and the rate of wound contraction calculated at the end of the study and the results are depicted in Table-1 and FIG. 1.

TABLE-US-00002 TABLE 1 Excision wound area - % Inhibition: Treatment Day Formulation 4 8 12 14 16 18 20 22 Chitosan in carbonated water 36.21 56.49 86.07 92.87 96.44 97.15 98.74 99.56 (Example-1) Chitosan Solution 27.62 49.38 86.21 92.64 98.39 98.74 99.14 99.65 (without carbonation) Carbonated water 40.30 54.20 80.20 92.99 95.48 96.83 97.97 99.02 Reference Savlon ® 24.37 37.83 71.18 83.69 95.47 96.90 98.90 99.73 Normal Control (Untreated) 28.23 40.59 71.52 91.59 95.83 96.23 97.79 99.17

Example-8: Procedure for Incision Wound Healing Study

[0105] Two parallel six cm paravertebral incisions were made through the full thickness of the skin, 1 cm lateral to the midline of vertebral column after giving anesthesia. Wounds were closed with interrupted sutures, 1 cm apart, with surgical suture. The given formulations were applied topically for 10 days. The sutures were removed on the 7.sup.th post-wounding day. Wound breaking strength was measured on the 10.sup.th post-wounding day in anaesthetized rats. Wound breaking strength was determined by calculating the weight required to break the wound and the results are depicted in Table-2 and FIG. 2.

TABLE-US-00003 TABLE 2 Incision-wound healing: Average Wound Break Composition Strength (Grams) Chitosan in carbonated water (Example-1) 398 Chitosan Solution (without carbonation) 314 Carbonated water 241 Reference Savlon ® 366 Normal Control (Untreated) 216

Examples 9 to 12

[0106] Spray Composition:

TABLE-US-00004 Example 9 Example 10 Example 11 Example 12 Ingredient (w/v) (w/v) (w/v) (w/v) Chitosan 0.1% 0.1% 0.1% 0.1% Lactic acid 0.5% 0.5% 1.2% 1.2% Carbonic acid 0.2% 0.3% 0.2% 0.3% Water Q.S to 100% Q.S to 100% Q.S to 100% Q.S to 100% pH 2.96  2.96  2.58  2.61 

[0107] Process for Preparation of Examples 9 to 12

[0108] Manufacturing Process A [0109] a. Collect 110% purified water required for the batch in the manufacturing vessel. [0110] b. Collect 90% of the purified water required for the batch in a separate vessel. Remaining quantity shall be used for volume make up. [0111] c. To the step b, add Lactic acid under stirring. [0112] d. To the step c, add Chitosan and stir until Chitosan is completely solubilized. [0113] e. Volume made-up to 100% of the batch size with purified water. [0114] f. Filter the solution through 0.2μ and cool to 2-10° C. [0115] g. Carbonate the solution and immediately fill into the Bag-On-Valve aerosol dispensing system.

[0116] Manufacturing Process B [0117] a. Collect 110% purified water required for the batch in the manufacturing vessel. [0118] b. Collect 90% of the purified water required for the batch in a separate vessel. Remaining quantity shall be used for volume make up. [0119] c. To the step b, add Chitosan under stirring to get homogenous dispersion. [0120] d. To the step c, add Lactic acid and stir until Chitosan is completely solubilized. [0121] e. Volume made-up to 100% of the batch size with purified water. [0122] f. Filter the solution through 0.2μ and cool to 2-10° C. [0123] g. Carbonate the solution and immediately fill into the Bag-On-Valve aerosol dispensing system.

Example 13: Description, pH, Carbonic Acid (g/L) and Degree of De-Acetylation of Chitosan Testing of Example 12

[0124] Composition of Example-12 was stored at 40° C.±2° C./75±5% RH and tested for description, pH, carbonic acid (g/L) and degree of de-acetylation of chitosan for a period of six months and the results are depicted in Table-3.

TABLE-US-00005 TABLE 3 Tests Carbonic Degree of De- acid content acetylation of Time pointy Description pH (g/L) Chitosan (%) Initial Clear colorless 2.61 3.08 86.2% solution 1 Month Clear colorless 2.57 3.02 85.5% solution 2 Months Clear colorless 2.52 2.97 86.3% solution 3 Months Clear colorless 2.58 2.99 87.1% solution 6 Months Clear colorless 2.52 3.06 86.4% solution

[0125] The above results indicate that the pH of the composition, carbonic acid content and degree of de-acetylation of chitosan is stable for a period of six months 40° C.±2° C./75±5% RH.

Example 13: Carbonic Acid Content Evaluation of Example-12 Composition when Stored in High-Pressure Resistant Container Bottle with a Nozzle (Can) and Bag-On-Valve (BOV) Aerosol Dispensing System

[0126] Carbonic acid content (g/L) of Example-12 filled in High-pressure resistant container bottle with nozzle system (having propellant) and Bag-On-Value (BOV) Aerosol dispensing system was evaluated for a period of 10 days and results are depicted in Table-4.

TABLE-US-00006 TABLE 4 Carbonic acid content (g/L) High-pressure resistant container Bag- On-Valve bottle with a nozzle system Aerosol (BOV) (containing composition of Example - dispensing Day 12 mixed with propellant) system Day 1 3.02 3.03 Day 2 2.99 3.01 Day 3 3.01 3.02 Day 4 2.96 2.99 Day 5 2.94 2.96 Day 6 2.90 2.97 Day 7 2.60 2.95 Day 8 2.45 2.94 Day 9 2.31 2.96 Day 10 2.15 2.91

[0127] The results indicate that the composition filled in BOV Aerosol dispensing system has better Carbonic acid content (g/L) throughout the usage period when compared to the composition filled in High-pressure resistant container bottle with a nozzle system (containing composition of Example-12 mixed with propellant).