TECHNICAL FIELD

The present invention relates to a herbal vulnerary and anti-microbial topical care composition useful in domestic animals and a process for preparing the same. The present invention provides a herbal composition which is synergistically effective in killing microbes and maggots, enhancing the wound healing activity and restoring the normal skin conditions. The composition of the present invention is effective in a variety of other skin disorders. Moreover the unique gel formulation provides the convenience of better stickability even on wet and weeping wounds.

BACKGROUND OF INVENTION

SKIN DISEASES, WOUNDS AND OTHER DERMATOLOGICAL DISORDERS

Skin diseases are of various types in domestic animals and they may be classified as primary and secondary skin diseases. Besides the skin diseases, wound also is quite common in domestic animals and wounds are classified according to the nature of effect produced viz., incised, punctured, lacerated and contused. Any one of these forms of wounds may become infected within pus forming organisms and can develop into a suppurative, septic wound. Since pus attracts flies, therefore fly menace cannot be ruled out and the flies lay egg on wound area leading to maggots. Infested tissue of wound area with larvae of dipterous flies is technically known as myiasis and larvae of blow flies, screw worm, flies, flesh flies, green flies burrow into the sub-cutaneous tissue and produce cutaneous myiasis resulting in wound with foul smelling discharge.

The destruction of tissue and its by products may cause toxemia. Animals become restless, irritated and feel pain. The animals rub the affected area against any object and bite the area. A large number of domestic animals are usually infested with microbials, the infection being prevalent throughout the year. The animals usually affected are the cows, buffaloes, horses, goats, sheeps and dogs. The diseases in livestock can affect the nutrition and economy of a family, a village or a nation. Hence, there is an important link between the health of livestock and the country's economy.

CONVENTIONAL AGENTS, CURRENT PRACTICES, DEMERITS

For treating various kinds of wounds in domestic animals, various measures and usage of products are being adopted. Some of them are as conventional as using of break oil / crude turpentine oil on the wound. Moreover some herbal products are also available of such products is not that high. Because of which the healing process of the wound is getting delayed. The cattle farmer who is generally poor and he keeps the animal for sustenance is not able to bear the high cost of treatment / application of such products for a very long period and therefore he ignores the treatment of wounds or results to the usage of conventional treatment which does not provide a holistic solution to the problem, resulting into further complicating the wound and that in turn directly affects the productivity of the animal.

Moreover it has been noticed that application of ointment on wet / weeping wound is very difficult and therefore in such cases, the wound healing further gets delayed because of poor / improper application of the ointment since the ointment does not stick properly on wet / weeping wound.

STATEMENT OF INVENTION

Accordingly the present invention provides a synergistic herbal composition comprising extracts of Pongamia glabra and Pinus longifoli, optionally along with pharmaceutically acceptable excipient(s); a process for preparation of synergistic herbal composition comprising extracts of Pongamia glabra and Pinus longifoli optionally along with pharmaceutical ly acceptable excipient(s), wherein said process comprises acts of a) obtaining extracts of Pongamia glabra and Pinus longifoli in suitable solvents respectively; b) mixing the extracts together and c) optionally adding the excipients(s) to obtain the synergistic herbal composition.

DETAILED DESCRIPTIONOF INVENTION

The present invention is in relation to a synergistic herbal composition comprising extracts of Pongamia glabra and Pinus longifoli, optionally along with pharmaceutical 1y acceptable excipient(s), wherein concentration of the Pongamia glabra extract is ranging from about 5% to about 45% w/w and concentration of the Pinus longifoli extract is ranging from about 10% to about 80% w/w.

In another embodiment of the present invention, pharmaceutically acceptable excipient is selected from a group comprising gelling agents, humectants, liquid paraffin, heavy mineral oil, binders, disintegrants, diluents, lubricants, plasticizers, permeation enhancers, solubilizers, preservatives and colouring agents or any combination thereof.

In still another embodiment of the present invention, concentration of the pharmaceutically acceptable excipient is ranging from about 5% to about 30% w/w.

In still another embodiment of the present invention, the gelling agent is selected from a group comprising fumed silicon carbide.

In still another embodiment of the present invention, concentration of the gelling agent is ranging from about 2% to about 12% w/w.

In still another embodiment of the present invention, the humectant is selected from a group comprising propylene glycol.

In still another embodiment of the present invention, concentration of the humectant is ranging from about 0.01% to about 2.5% w/w.

In still another embodiment of the present invention, said composition is formulated into a form selected from a group comprising gel, lotion, cream, ointment, aerosol spray, aqueous or oily suspensions, dispersible powders and phytoceuticals.

The present invention is also in relation to a process for preparation of synergistic herbal composition comprising extracts of Pongamia glabra and Pinus longifoli optionally along with pharmaceutical^ acceptable excipient(s), wherein said process comprises acts of

a) obtaining extracts of Pongamia glabra and Pinus longifoli;

b) mixing the extracts together; and

c) optionally adding the excipients(s) to obtain the synergistic herbal composition.

In still another embodiment of the present invention, the extracts are in form of oil.

A product which is having a very high anti-microbial effect, vulnerary effect fly repellent effect and with good stickability even on wet wound is needed for treating various kinds of skin lesions and disorders in domestic animals.

Globally, there is a widespread demand for natural products based on herbs. Since many herbs are known to be effective in skin diseases, the present invention is based on herbs.

Based on the above description, the ideal anti-microbial, vulnerary product with good stickability should have the following essential features.

1. Easy application on wet/weeping wounds (user friendly).

2. Wide spectrum anti-microbial property.

3. Better transdermal absorption.

4. Enhance wound-healing process.

5. Effective in treating all kinds of wounds viz., incised, punctured, lacerated, contused, septic, aseptic, wide spread lesion, footrot etc.,

6. Better spreadabilities.

7. Good fly-repellent property.

After understanding the requirement in connection with the development, especially the domestic animal industry, it was learnt that efficacy of the various types of anti-microbial products hitherto available differs.

The present invention has been developed to meet the above said requirements.

Another embodiment of the present invention is to provide a product, which is also having anti-bacterial, anti-fungal, anti-microbial and wound healing properties.

The embodiment of the present invention resides in the selection of the constituents of this herbal vulnerary and anti-microbial topical care composition.

Another embodiment of the present invention, lies in the selection of the ingredients and also in the proportions selected in such a manner so as to exhibit maximum synergistic activity, which is unique to this product.

Another embodiment of the present invention is to provide an improved herbal vulnerary and anti-microbial topical care composition for domestic animals.

Another embodiment of the present invention is to provide an improved herbal vulnerary and anti-microbial topical care composition for domestic animals, which is very potent, nontoxic and safe.

Yet another embodiment of the present invention is to provide an improved herbal vulnerary and anti-microbial topical care composition useful for domestic animals which posses fly repellent property.

Another embodiment of the present invention is to provide an improved herbal vulnerary and anti-microbial topical care composition useful for domestic animals, which enhance the wound healing process.

Yet another embodiment of the present invention is to provide an improved herbal vulnerary and anti-microbial topical care composition useful for domestic animals, which has better spreadabilities and sticks well even on wet / weeping wound.

Still another embodiment of the present invention is to provide an improved herbal vulnerary and anti-microbial topical care composition useful for domestic animals which kills maggots and prevents further re-infestation.

Yet another embodiment of the present invention is to provide an improved herbal vulnerary and anti-microbial topical care composition useful for domestic animals which is effective in treating all kinds of wounds including deep seated FMD foot lesions, wide spread lesions and footrot, incised wound, lacerated wound, contused wound, punctured wound, septic wound etc.

Another embodiment of the present invention is to provide an improved herbal vulnerary and anti-microbial topical care composition useful for domestic animals which is non toxic to the animals and its human handlers.

The present invention also relates to a process for the preparation of a herbal vulnerary and anti-microbial topical care composition.

The other embodiment of the invention resides in the use of such a novel composition in the form of gel, lotion, ointment, cream, aerosol spray and the like.

The said composition can also be mixed with the bases of gel, cream, lotion or ointment etc to obtain the desired dosage form without much change in its efficacy.

Ayurveda (Indian system of medicine) recommends several medicinal plants useful for skin and its related disorders. Ayurvedic literature like Bhavaprakasha Nighantu,

Yogaratnakara, Vaidyachinthamani and Charakasamhita has references to herbs capable of being used in the treatment of skin disorders. The present invention is based on our experiments and knowledge about the medicinal properties of various herbs along with our studies on traditional literature.

In an attempt to develop a potent, safe, effective herbal vulnerary and anti-microbial topical care composition more than 20 Indian medicinal plants were selected for the screening based on their reports of wound healing, anti-microbial, anti-fungal activities and their significance in ethnomedicine. The selected herbs were coarsely powdered and extracted / distilled with suitable solvents using the conventional methods of extraction / distillation. The extracts and distillates thus obtained were subjected to a battery of experimental studies. The plant substances, which showed promising activity in these studies, were short-listed further for developing prototype compositions based on the Ayurvedic principles and modern concepts of synergism.

The compositions prepared were reanalyzed in experimental and field trials for their efficacy. Based on these results a particular prototype comprising of Pinus longifolia and Pongamia glabra was showed good vulnerary and anti-microbial activity. Even though all the above said plants have been known in Ayurveda but the synergistic activity of their extracts / distillates in a particular combination having vulnerary, anti-microbial activities is hither to not known and hence is novel.

The novelty of the present invention is present in the careful selection and blending of extracts and distillates obtained from plants like Pongamia and Pinus. The extracts and distillates were blended in a series of specific proportions based on the concept of synergy and several compositions were prepared. Each of these compositions was then subjected to evaluation for anti-microbial activity and wound healing using both the in vitro and in vivo animal models.

The most potent composition was selected and subjected to further clinical evaluation in cows and buffaloes having microbial wounds. The efficacy of the selected composition in the target species was found to be very encouraging. Further trials were conducted to recheck the efficacy of this novel composition in different forms like gel, lotion etc. This composition can be used to treat all types of wounds, dermatomycosis, ringworm and mango scabies. This composition has been mainly developed to treat myiasis in cattle.

The composition has potent anti-microbial, maggoticidal and has excellent wound healing or vulnerary activity.

Accordingly the present invention provides a herbal vulnerary and anti-microbial topical care composition comprising:

(a) 5 to 45% w/w of the total weight of the composition, of the distillates of Pongamia glabra

(b) 10 to 80% w/w of the total weight of the composition, of the distillates of Pinus longifolia

(c) 5 to 30% w/w of the total weight of the composition of the excipients

(d) 0.01 to 2.5% w/w of the total weight of the composition, of the humectant

(e) 2 to 12% w/w of the total weight of the composition, of the gelling agent

And mixing 1 & 2 thoroughly in a suitable vessel and adding 3, 4, 5 to it and stir thoroughly for 15 - 20 min to get the final product in the form of gel.

According to another feature of the present invention the composition may be formed into different dosage forms like lotion, ointment, aerosol spray etc.

For facilitating making the composition in the above dosage forms, pharmaceutical excipients which are usually employed to prepare such dosage forms may be employed.

By way of examples excipients like propylene glycol, liquid paraffin and fumed silicon
dioxide etc., are a mixture there of may be used.

According to yet another feature of the present invention there is provided a process for preparing the herbal vulnerary and anti-microbial topical care composition, which comprises mixing a first mixture of

1. 5 to 45% w/w of the extracts / distillate of Pongamia

2. 10 to 80% w/w of the distillate of Pinus and mixing the resultant first mixture with

3. 5% to 3 0% w/w of excipient

4. 2 to 12% w/w of gelling agent

5. 0.01 to 2.5% w/w of humectant
in a closed stainless steel container at atmospheric pressure and at room temperature and stirring the composition in the said container for a sufficient time to obtain a homogenous composition to result in the final product in the form of gel.

According to another feature of the present invention the composition containing heavy mineral oil may be used as an excipient.

According to another feature of the present invention the composition containing propylene glycol may be used as a humectant.

According to another feature of the present invention the composition containing silicon
dioxide may be used as a gelling agent.

According to another feature of the present invention the composition may be formed into different dosage forms like lotion, ointment, aerosol spray etc.

For facilitating making the composition in the above dosage forms, pharmaceutical excipients which are usually employed to prepare such dosage forms may be employed.

Excipients like propylene glycol, heavy mineral oil and propylene glycol or a mixture thereof may be used.

The invention is described in detail in the examples given below which are provided by way of illustration only and therefore should not be construed to limit the scope of the invention.

PREPARATION OF EXAMPLES 1,2 & 3 EXAMPLE 1

The preparation of herbal vulnerary and anti-microbial topical care composition comprises:

(a) Mixing a first mixture of

1. 23% w/w of the extract/distillate of Pongamia

2. 53% w/w of the extract/distillate of Pinus and mixing the resultant first mixture with

(b) a second mixture of

3. 10% w/w of excipient (heavy mineral oil)

4. 12% w/w of gelling agent (Fumed silicon dioxide) and

5. 2%w/w of humectant (Propylene glycol)

The 1 & 2 is thoroughly mixed in a suitable vessel and then added to second mixture of 3,
4 and 5 and stiring both the mixtures thoroughly for 15 - 20 min to get the final product in the form of gel. This composition is henceforth called as NR-TG-A.

EXAMPLE 2

The preparation of herbal vulnerary and anti-microbial topical care composition comprises:

(a) Mixing a first mixture of

1. 8 % w/w of the extract/distillate of Pongamia

2. 62 % w/w of the extract/distillate of Pinus and mixing the resultant first mixture with

(b) a second mixture of

3. 20% w/w of excipient (heavy mineral oil)

4. 8% w/w of gelling agent (Fumed silicon dioxide) and

5. 2%w/w of humectant (Propylene glycol)

The 1 & 2 is thoroughly mixed in a suitable vessel and then added to second mixture of 3,
4 and 5 and stiring both the mixtures thoroughly for 15 - 20 min to get the final product in the form of gel. This composition is henceforth called as NR-TG-B.

EXAMPLE 3

The preparation of herbal vulnerary and anti-microbial topical care composition comprises: (a) Mixing a first mixture of

1. 18% w/w of the distillate of Pongamia

2. 58% w/w of the distillate of Pinus
and mixing the resultant first mixture with

(b) a second mixture of

3. 12% w/w of excipient (heavy mineral oil)

4. 10% w/w of gelling agent (Fumed silicon dioxide) and

5. 2%w/w of humectant (Propylene glycol)

The 1 & 2 is thoroughly mixed in a suitable vessel and then added to second mixture of 3, 4 and 5 and stiring both the mixtures thoroughly for 15 - 20 min to get the final product in the form of gel. This composition is henceforth called as NR-TG-C.

Each of the above said examples were subjected to the following experiments to select the best comparative wound healing activity studies of compositions described in example 1, 2 & 3.

TABLE 1: MIC of Karanj oil, Pine Oil and prototypes
EFFICACY STUDY 1:

OBJECTIVE: Evaluation of wound healing activity of the present invention NR-TG-A, B, C in excised wound model in rats.

The study was conducted on rats. The prototypes of the present inventions were evaluated in rats. They were applied topically on the excision wound made on the dorsal surface of the animal. All the three groups showed significant increase in granulation tissue weight,
collagen and hydroxy proline content compared to the control group. The glucosamine
content was also found to be comparatively higher in the treatment groups.

RESULTS:

TABLE 2: WOUND HEALING ACTIVITY OF NR-TG-A, B, C IN EXCISION WOUND IN RATS

* p < 0.05 compared to control
* p < 0.05 compared to Gel base

Thus the increase in collagen, hydroxy proline and glucosamine of granulation tissue in the present study provide evidence to the increased tensile strength of the wound, which was proven by the incised wound model of NR-TG-A, B. In-house pharmacology report No. PCL/WH/351

EFFICACY STUDY 2:

OBJECTIVE: EVALUATION OF ANTI-MICROBIAL ACTIVITY OF THE PRESENT NR-TG-A, B, C

Skin infection is a common manifestation among cattle, goat, sheep and other animals. Bacterial and fungal agents play a predominant role in infectious conditions. Important bacterial infections include pyoderma, impetigo etc. Bacterial infections are commonly caused by Staphylococcus aureus[l] and Pseudomonas sps. The fungal infections are caused by dermatophytes like Trichophyton sps., Epidermophyton sps., Microsporum sps.and Candida sps that commonly cause skin diseases of the people and animals. The present invention is a topical antiseptic gel developed by the inventor.

The present invention is indicated in topical infectious conditions in cattle, sheep, goat etc. The present study is designed to evaluate the anti-microbial activity (in vitro) of the present invention, NR-TG-A, B, C.

RESULTS:
The results are presented in Table 4. It is found to be most effective against S. aureus and P. vulgaris

TABLE 4: MINIMUM INHIBITORY CONCENTRATION (MIC) OF THE PRESENT INVENTION.

CONCLUSION: The present invention coded as NR-TG-A exhibits a broad spectrum antimicrobial effect, which substantiates its use as a topical anti-microbial.

Based on this study all prototypes appear to be exhibiting potential anti microbial activity.

EFFICACY STUDY 3:

Comparative evaluation of NR/TG/A, B and C gel in the management of surgical and other types of wounds in cattle and buffaloes.
OBJECTIVES: This research work was done with the following objectives.

1. To evaluate the healing effects of NR/TG/A, B, C gel in surgical wounds.

2. To assess the efficacy of these gels in other types of wounds viz., microbial, non-surgical and general wounds.

3. To study the histopathological response after application of these gels in experimentally created surgical wounds.

4. To estimate the collagen, elastin, hydroxyproline and hexosamine contents in the healing tissues in the experimentally induced surgical wounds.

APPLICATION: The application of the gels was twice daily till complete healing. The
surgically induced experimental wounds were protected with gauze thane cloth at the dorso-lumbar region.

RESULTS:

I. FIELD OBSERVATIONS:

On the basis of oozing of exudates, degree of swelling, pain at the site, appearance of granulation tissue and contraction of wound, various surgical wounds were evaluated after application of these three gels (NR-TG-A, B, C).

The swelling was maximum up to 3 days in NR-TG-C followed by 5 days in NR-TG-B and 7 days in NR-TG-B and A respectively. The pain at the wound site was noted up to 5 days in the group of animals applied with NR-TG-C and up to 7 and 8 days in NR-TG-B and A treated wounds respectively.
Table 5: Showing efficacy of various prototypes NR-TG-A, B and C for the management of various types of wounds (surgical, microbial and general wounds) in cattle and buffaloes.

H1STOPATHOLOGICAL STUDIES:

The histopathological studies were concluded in 9 animals in which the surgical wounds were created experimentally. A total of 54 full thickness wounds of size 3 X 2 cm were created on either side of the dorso-lumbar region (6 wounds / animal) under xylazine sedation (15 mg) and local infiltration with 2% lignocaine hydrochloride.

Out of 54 full thickness wounds, 12 wounds were treated with NR-TG-A, 12 with NR-
TG-B and 12 with NR-TG-C where as remaining 18 wounds were kept as untreated control. The wounds treated with NR-TG-C showed swelling and exudation up to 3 days and pain up to 5 days. NR-TG-B treated wounds showed swelling and exudation up to 5 days and pain up to 7 days. The wounds treated with NR-TG-A showed swelling and exudation up to 7 days and pain persisted up to 8 days.

CONCLUSIONS:

On the basis of the various findings of this research project, the following conclusions were drawn.

1. NR-TG-gel A, B and C are very safe and effective for post-operative wound dressing in cattle and buffaloes.

2. The clinical wound healing occurred in 18 days, 15 days and 12 days in wounds treated with NR-TG-A, B and C respectively. Thus based on number of days required for healing, NR-TG-C was the best followed by NR-TG-B and C respectively.

3. In surgical wounds, where the post-operative dressing was done by these gels, uneventful healing in all the cases occurred in NR-TG-C treated animals. Infection in three cases was reported in NR-TG-B treated animals. Infection in five cases was reported in animals dressed with NR-TG-A. The healing time was also minimal in NR-TG-C treated cases followed by NR-TG-B and NR-TG-A.

4. These gels can also be used for the treatment of septic wounds in combination with antibiotics.

5. Clinical observations post application of these gels in induced surgical wounds that swelling and exudation were noted up to 3rd day and pain up to 5th day in wounds treated with NR-TG-C where as in NR-TG-B, treated wounds swelling and exudation persisted up to 5 days and pain up to 7 days. In wounds treated with NR-TG-A, swelling and exudation persisted up to 7 days and pain up to 8 days. Thus NR-TG-C was again proved to be more efficacious in minimizing the inflammatory response at the wound site in comparison to other gels.

6. Histopathological examination of the healing tissue treated with three gels revealed significantly higher rate of healing in comparison to other gels.

7. Histobiochemical studies revealed significant and progressive increase in the collagen contents, hydroxyproline, elastin and significant decrease in hexosamine in wounds treated with three gels in comparison to control wounds. Higher contents of collagen, hydroxyproline, elastin and low concentration of hexosamine in wounds treated with NR-TG-C further confirmed the supremacy of this NR-TG-C over other gels.

8. All the three gels have excellent fly repellent property. They were equally efficacious in microbial wounds also.

9. The tissue spreadability of NR-TG-C is excellent although NR-TG-B and A were also well admissible with the tissue. Further the fragrance of the NR-TG-C is also better than other two gels.

10. Finally, based on the detailed study on the various aspects of the wound healing in response to treatment with these three gels, these are graded as follows. NR-TG-C is excellent wound healer followed by NR-TG-B and NR-TG-A, which also have got good wound healing properties. Gel B and A did not differ significantly amongst each other.

It would be very clear and undoubtedly confirmed that the composition of the present invention, which is the result of specific and unique combination of the specific ingredients, which combination imparts unique synergistic vulnerary anti-microbial anti-bacterial and wound healing activities to the resulting composition thereby imparting the novelty to the composition.

ADVANTAGES OF THE PRESENT INVENTION:

1. Easy application on wet/weeping wounds (user friendly).

2. Wide spectrum anti-microbial property.

3. Advanced wound-healing process with better transdermal absorption.

4. Enhance wound-healing process.

5. Effective in treating all kinds of wounds viz., incised, punctured, lacerated, contused, septic, aseptic, wide spread lesion, footrot etc.,

6. Better spreadabilities.

7. Good fly-repellent property.

I CLAIM:

1) A synergistic herbal composition comprising extracts of Pongamia glabra and Pinus longifoli, optionally along with pharmaceutically acceptable excipient(s).

2) The synergistic herbal composition as claimed in claim 1, wherein concentration of the Pongamia glabra extract is ranging from about 5% to about 45% w/w and concentration of the Pinus longifoli extract is ranging from about 10% to about 80% w/w.

3) The synergistic herbal composition as claimed in claim 1, wherein pharmaceutically acceptable excipient is selected from a group comprising gelling agents, humectants, liquid paraffin, heavy mineral oil, binders, disintegrants, diluents, lubricants, plasticizers, permeation enhancers, solubilizers, preservatives and colouring agents or any combination thereof.

4) The synergistic herbal composition as claimed in claim 3, wherein concentration of the pharmaceutically acceptable excipient is ranging from about 5% to about 30% w/w.

5) The synergistic herbal composition as claimed in claim 3, wherein the gelling agent is selected from a group comprising fumed silicon carbide.

6) The synergistic herbal composition as claimed in claim 5, wherein concentration of the gelling agent is ranging from about 2% to about 12% w/w.

7) The synergistic herbal composition as claimed in claim 3, wherein the humectant is selected from a group comprising propylene glycol.

8) The synergistic herbal composition as claimed in claim 7, wherein concentration of the humectant is ranging from about 0.01% to about 2.5% w/w.

9) The synergistic herbal composition as claimed in claim 3, wherein said composition is formulated into a form selected from a group comprising gel, lotion, cream, ointment, aerosol spray, aqueous or oily suspensions, dispersible powders and phytoceuticals.

10) A process for preparation of synergistic herbal composition comprising extracts of Pongamia glabra and Pinus longifoli optionally along with pharmaceutically acceptable excipient(s), wherein said process comprises acts of

d) obtaining extracts of Pongamia glabra and Pinus longifoli in a suitable solvent;

e) mixing the extracts together; and

f) optionally adding the excipients(s) to obtain the synergistic herbal composition.

11)The process for preparation of synergistic herbal composition as claimed in claim 10, wherein the extracts are in form of oil.