“A POLYHERBAL FILM FORMING SPRAY FORMULATION AND PREPARATION METHOD THEREOF”

Abstract

“A POLYHERBAL FILM FORMING SPRAY FORMULATION AND PREPARATION METHOD THEREOF”. ABSTRACT The present invention relates to a polyherbal film forming spray formulation for wound healing. The polyherbal film forming spray comprises extract of Aegle Marmelos, Ocimum Basilicum and Tabernaemontana divaricata along with pharmaceutically inert excipients. The invention further provides method of preparation of a polyherbal film forming spray for wound healing.

Specification

Description:FORM 2
THE PATENTS ACT 1970
(39 of 1970)
&
The Patent Rules 2003
COMPLETE SPECIFICATION
(See sections 10 & rule 13)
1. TITLE OF THE INVENTION
"A POLYHERBAL FILM FORMING SPRAY FORMULATION AND PREPARATION METHOD THEREOF"
2. APPLICANT (S)
NAME NATIONALITY ADDRESS
Iniya Madhan Kumaar Indian NCRD's Sterling Institute of Pharmacy, Plot No. 93/93A, Sector 19, Near Seawoods Railway Station, Nerul (E), Navi Mumbai - 400706, Maharashtra, India.
3. PREAMBLE TO THE DESCRIPTION
COMPLETE SPECIFICATION
The following specification particularly describes the invention and the manner in which it is to be performed.





FIELD OF INVENTION
The present invention in general relates to a polyherbal film forming spray formulation for wound healing. Polyherbal film forming spray comprises extract of Aegle Marmelos, extract of Ocimum Basilicum, and extract of Tabernaemontana divaricata along with pharmaceutically acceptable inert excipients. The invention further provides preparation method of polyherbal film forming spray formulation.

BACKGROUND OF INVENTION
Wounds create openings in the skin, allowing bacteria, viruses, and other pathogens to enter the body. Proper wound care is essential to clean the wound, remove debris and foreign material, and minimize the risk of infection. Effective wound care creates an optimal environment for the body's natural healing processes, including keeping the wound clean and moist to facilitate new tissue growth and speed up healing. Without proper care, wounds can develop complications such as infection, delayed healing, excessive scarring, or tissue damage. Consistent wound care helps reduce the likelihood of these complications and promotes better healing outcomes.

Wound healing is a complex and dynamic process that the body undergoes to repair damaged tissue. This process involves several phases, each characterized by distinct cellular and molecular activities. Understanding wound healing is essential for developing effective treatments and interventions to promote recovery and prevent complications. The primary phases of wound healing are hemostasis, inflammation, proliferation, and remodelling.

Hemostasis is the initial response to a wound and begins immediately after injury. Its main goal is to stop bleeding. When a blood vessel is damaged, it constricts to reduce blood flow. Platelets, small blood cells, adhere to the exposed collagen of the damaged vessel wall, forming a temporary "platelet plug." This aggregation activates the coagulation cascade, a series of reactions that result in the formation of fibrin, a protein that weaves through the platelet plug to stabilize it and form a more durable blood clot.

The inflammation phase follows hemostasis and typically lasts for a few days. This phase is crucial for preventing infection and clearing away debris. It is characterized by redness, heat, swelling, and pain. During this phase blood vessels expand, increasing blood flow to the injured area, bringing in immune cells, nutrients, and oxygen. Neutrophils and macrophages, types of white blood cells, migrate to the wound site to engulf and digest bacteria, dead cells, and other debris. Immune cells release signalling molecules called cytokines, which coordinate the inflammatory response and attract more immune cells to the wound.

The proliferation phase involves the formation of new tissue and typically lasts several weeks. It includes angiogenesis the formation of new blood vessels to supply the wound with oxygen and nutrients. Fibroplasia in which fibroblasts, cells that produce collagen, proliferate, and synthesize extracellular matrix components, creating a new tissue framework. The wound bed fills with granulation tissue, which is rich in blood vessels and collagen, providing the foundation for new skin. Afterward, re-epithelialization occurs, where epithelial cells move across the wound bed to form new skin.

The remodelling, or maturation, phase can last from months to years, during which the initially disorganized collagen fibers are rearranged, cross-linked, and aligned along tension lines to enhance the tensile strength of the new tissue. Concurrently, the excessive blood vessels formed during proliferation regress, leading to a paler scar tissue. Over time, the new tissue matures into a scar, which, although generally less flexible and weaker than the original tissue, undergoes continuous refinement to maximize the functional and aesthetic recovery of the wound.

Conventional wound management often relies on the use of dressings and topical agents, which may have limitations in terms of efficacy, patient comfort, and ease of application. Moreover, the rise of antimicrobial resistance underscores the need for alternative approaches to wound care that are both effective and sustainable.

In recent years, there has been growing interest in herbal medicine as a source of novel therapeutics for wound healing. Herbal extracts contain a diverse array of bioactive compounds with demonstrated antimicrobial, anti-inflammatory, and tissue-regenerating properties. These natural compounds offer a promising avenue for developing effective wound care solutions that can address the multifaceted challenges of wound management.

However, while herbal extracts hold great potential for wound healing, their application in clinical practice is often hindered by practical challenges such as formulation stability, bioavailability, and ease of use. Incorporating herbal extracts into conventional wound care products, such as dressings or topical solutions, may not fully harness their therapeutic benefits or may present challenges in terms of product stability and efficacy.

At the same time, advancements in wound care technology have led to the development of novel formulations that can create protective barriers over wounds while facilitating the healing process. Film-forming sprays represent one such innovation, offering a convenient and effective way to protect wounds from external contaminants while maintaining an optimal moisture balance conducive to healing.

Film-forming sprays for wound healing are advanced topical treatments designed to create a protective barrier over a wound. These formulations include polymers which form the basis of the film and antimicrobial agents to prevent infection. These sprays form a thin, flexible film when applied to the skin, offering several benefits that aid in the healing process. The spray is applied directly to the wound area, where it rapidly dries to form a continuous, breathable film. The film conforms to the contours of the wound, providing a snug fit that protects against contaminants and friction.

The film acts as a physical barrier, protecting the wound from bacterial contamination and external irritants. It helps maintain an optimal moisture balance, preventing the wound from drying out, which is crucial for effective healing. These sprays form a flexible film that moves with the skin, making them comfortable for patients and suitable for areas with high movement. The spray format allows for easy and uniform application, even on irregular wound surfaces or hard-to-reach areas. The transparent nature of many films allows for ongoing visual assessment of the wound without removing the protective layer.

Given the complementary nature of herbal medicine and modern wound care technology, there is a compelling opportunity to integrate herbal extracts into film-forming spray formulations for enhanced wound healing outcomes. By leveraging the synergistic effects of herbal extracts with the protective benefits of film-forming technology, it is possible to develop a novel wound care solution that addresses the unmet needs of patients and healthcare providers alike.

An article entitled, 'Formulation, characterization and in-vivo evaluation of standardized Tabernaemontana divericata extract hydrogel for wound healing' by Nupur Gargate, Sadhana Raut, Harshad Kapare, Poonam Shende, Ritesh Bhole in Journal of Ayurveda and Integrative Medicine 2024, Volume 15, Issue 3, discloses formulation and evaluation of wound healing potential of standardized ethanolic extract of Tabernaemontana divericata leaves hydrogel on experimental models of wounds in Wistar rats with the design of experiment approach.

An article entitled, 'Synthesis of chemically cross-linked hydrogel films based on basil seed (Ocimum basilicum L.) mucilage for wound dressing drug delivery applications.' by Masoomeh Sadat Hosseini, Mohammad Reza Nabid in International Journal of Biological Macromolecules 2020, Volume 163, Pages 336-347 discloses preparation of pH-sensitive hydrogel films based on basil seed mucilage (OBM) biopolymer as a novel drug delivery system for wound dressing.

An article entitled, 'Physico-chemical, mechanical, and electrical performance of bael fruit gum-chitosan IPN films' by Manish Jindal, Vineet Kumar, Vikas Rana, A.K. Tiwary in Food Hydrocolloids Volume 30, Issue 1, January 2013, Pages 192-199 discloses gum obtained from unripe fruits of Aegle marmelos was co-processed with chitosan to improve the film forming property of the former.

There is still a need for a polyherbal film-forming sprays to bridge the gap between traditional herbal medicine and modern wound care by providing a combination of therapeutic properties of herbal extracts with the convenience and efficacy of film-forming technology. By harnessing the power of nature and innovation, this invention has the potential to revolutionize wound care and improve patient outcomes across a wide range of wound types and clinical settings.

OBJECTS OF THE INVENTION
Main object of the present invention is to provide a polyherbal film forming spray for wound healing and antimicrobial activity.

Another object of the present invention is to provide a polyherbal film forming spray comprising extract of Aegle Marmelos, Ocimum Basilicum, and Tabernaemontana divaricata along with pharmaceutically acceptable inert excipients.

Yet another object of the present invention is to provide a process for preparation of a polyherbal film forming spray.

SUMMARY OF THE INVENTION
In an embodiment, the present invention relates to a polyherbal film forming spray formulation comprising,
a) Extract of Aegle Marmelos;
b) Extract of Ocimum Basilicum;
c) Extract of Tabernaemontana divaricata; and
d) Pharmaceutically acceptable inert excipients.

In an aspect of the embodiment, the pharmaceutically acceptable inert excipients are film forming agent, plasticizer, permeation enhancer, and solvent.

In another aspect of the embodiment, the film forming agent is Hydroxypropyl methylcellulose.

In another aspect of the embodiment, the plasticizers are Propylene glycol and polyethylene glycol.

In another aspect of the embodiment, the permeation enhancer is Eucalyptus oil.

In another aspect of the embodiment, the solvent is Ethanol.

In another embodiment, the polyherbal film forming spray formulation comprising,
a) 4% v/v of mixture of extract of Aegle Marmelos, Ocimum Basilicum, and Tabernaemontana divaricata;
b) 2% w/v of Hydroxypropyl methylcellulose;
c) 2% v/v of Propylene glycol and Polyethylene glycol;
d) 1% v/v of Eucalyptus oil;
e) Ethanol,
wherein the percentages are with respect to the total weight of the formulation.

In another embodiment, the mixture of extract of Aegle Marmelos, Ocimum Basilicum and Tabernaemontana divaricata are present in the weight ratio of 1:1:1.

In another aspect of embodiment, the propylene glycol and polyethylene glycol are present in the weight ratio of 1:1.

In another embodiment, the present invention relates to a process for preparation of polyherbal film forming spray formulation comprises the steps of;
a) Dissolving hydroxypropyl methylcellulose into ethanol to form solution I;
b) Dissolving the extracts of Aegle Marmelos, Ocimum Basilicum, and Tabernaemontana divaricata in ethanol to form a solution II;
c) Mixing the solution I and solution II to form solution III;
d) Adding propylene glycol, polyethylene glycol and Eucalyptus oil to solution III to obtain the polyherbal film-forming spray formulation.

DETAILED DESCRIPTION OF THE INVENTION:
The present invention is all about a polyherbal film forming spray comprising,
a) Extract of Aegle Marmelos;
b) Extract of Ocimum Basilicum;
c) Extract of Tabernaemontana divaricata; and
d) Pharmaceutically acceptable inert excipients.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains.

The term "comprising", which is synonymous with "including", "containing", or "characterized by" here is defined as being inclusive or open-ended, and does not exclude additional, unrecited elements or method steps, unless the context clearly requires otherwise.

The term "a" and "an" refers to one or to more than one (i.e., to at least one) of the grammatical object of the article. The information provided in this document, and particularly the specific details of the described exemplary aspects, is provided primarily for clearness of understanding and no unnecessary limitations are to be understood from there.

As used herein, the term "about" means that the numerical value is approximate and small variations would not significantly affect the practice of the disclosed embodiments. Where a numerical limitation is used, unless indicated otherwise by the context, "about" means the numerical value can vary by ±10% and remain within the scope of the disclosed embodiments.

The term "Polyherbal formulation" refers to a medicinal product or preparation that contains a combination of multiple herbs or herbal extracts. Rather than relying on a single herb, polyherbal formulations harness the synergistic effects of various herbs to achieve therapeutic outcomes. These formulations are designed to address complex health conditions or to provide comprehensive health benefits by leveraging the diverse bioactive compounds present in different herbs.

The term "Film- forming spray formulation" refers to a specialized type of topical product designed to create a thin, protective film over the surface of the skin or wounds when sprayed. This formulation typically consists of ingredients that, when sprayed onto the desired area, undergo a rapid drying process to form a flexible and transparent film. This film adheres to the skin or wound bed, providing a barrier that protects against external contaminants, promotes wound healing, and maintains an optimal environment for tissue regeneration.

The term "pharmaceutically acceptable inert excipients", denotes any of the components of a cosmetic formulation other than the active and which are approved by regulatory authorities or are generally 'regarded as safe' for human use. A combination of excipients may also be used. The amount of excipient(s) employed will depend upon how much active agent is to be used. One excipient can perform more than one function.

Although the invention has been described with reference to specific embodiments, this description is not meant to be construed in a limiting sense. Various modifications of the disclosed embodiments, as well as alternate embodiments of the invention, will become apparent to persons skilled in the art upon reference to the description of the invention. It is therefore contemplated that such modifications can be made without departing from the spirit or scope of the present invention as defined.

Main embodiment of the present invention relates to a polyherbal film forming spray formulation comprising,
a) Extract of Aegle Marmelos;
b) Extract of Ocimum Basilicum;
c) Extract of Tabernaemontana divaricata; and
d) Pharmaceutically acceptable inert excipients.

Aegle marmelos, commonly known as bael or Bengal quince, is a species of tree native to the Indian subcontinent and Southeast Asia. The biological source of Aegle marmelos includes various parts of the tree such as the fruit, leaves, roots, and bark, each of which has been traditionally used in different medicinal and therapeutic applications.

Aegle marmelos as an antimicrobial agent in a film-forming spray for wound healing leverages its natural bioactive compounds, which exhibit significant antimicrobial properties. Aegle marmelos contains a variety of bioactive compounds that contribute to its antimicrobial activity, including alkaloids like aegeline and aegelinine, which have demonstrated antibacterial and antifungal properties. Tannins such as tannic acid and phlobatannins are known for their antimicrobial effects, while flavonoids like rutin and quercetin offer broad-spectrum antimicrobial activities. Additionally, coumarins such as marmin and imperatorin, along with essential oils like limonene and citral, exhibit significant antimicrobial properties against a range of pathogens.

Ocimum basilicum, commonly known as basil, is an herbaceous plant belonging to the family Lamiaceae. It is widely cultivated for its aromatic leaves, which are used in culinary, medicinal, and ornamental applications. Utilizing Ocimum basilicum as an antimicrobial agent in a film-forming spray for wound healing capitalizes on its inherent bioactive compounds, renowned for their potent antimicrobial effects. Ocimum basilicum harbours a range of bioactive compounds, prominently including essential oils abundant in constituents such as linalool, eugenol, and methyl chavicol. These constituents have been extensively studied and proven to exhibit antibacterial, antifungal, and antiviral properties, rendering them invaluable for addressing microbial infections and fostering the wound healing process.

The bioactive compounds derived from Ocimum basilicum provide effective antimicrobial protection by inhibiting microbial colonization of the wound, thereby lowering the risk of infection, and facilitating the healing process. Additionally, basil compounds exhibit anti-inflammatory properties, which contribute to the reduction of inflammation at the wound site, fostering an environment conducive to healing. Moreover, the antioxidant properties of basil compounds help shield cells from oxidative damage, promoting expedited wound closure and tissue regeneration.

Tabernaemontana divaricata, commonly known as crepe jasmine or pinwheel flower, is a species of flowering plant in the family Apocynaceae. Incorporating Tabernaemontana divaricata as an antimicrobial agent in a film-forming spray for wound healing capitalizes on its natural bioactive compounds, which exhibit potential antimicrobial properties. Extract of Tabernaemontana divaricata contains various chemical constituents, including alkaloids, flavonoids, and glycosides, some of which have antimicrobial activity.

Tabernaemontana divaricata contains alkaloids such as conophylline and voacangine, which have demonstrated antimicrobial activity against various pathogens, including bacteria and fungi. The bioactive compounds from Tabernaemontana divaricata can help prevent microbial colonization of the wound, reducing the risk of infection and supporting the healing process. Tabernaemontana divaricata shows promise as a potential antimicrobial agent in film-forming sprays for wound healing.

In another aspect of the embodiment, the pharmaceutically acceptable inert excipients are film forming agent, plasticizers, permeation enhancer and solvent.

The formulation includes film forming agent. Film forming agent are substances that help create a continuous, uniform film on a surface when the spray is applied. These agents are crucial in various industries, including cosmetics, pharmaceuticals, agriculture, and coatings. Film forming agent used in polyherbal film forming spray can be selected from Ethyl Cellulose, Hydroxyethyl Cellulose, Hydroxypropyl Methylcellulose, Eudragit, and Polyvinyl Alcohol.

In another aspect of the embodiment, the film forming is Hydroxypropyl Methylcellulose in a concentration of 2% w/v.

Hydroxypropyl methylcellulose (HPMC) is a commonly used film-forming agent in pharmaceuticals due to its versatile properties and wide range of applications. Hydroxypropyl methylcellulose (HPMC) stands out in pharmaceutical applications due to its non-toxic, biodegradable nature, and excellent compatibility with the human body, making it ideal for both oral and topical formulations. Its versatility allows for standalone use or combination with other polymers, catering to diverse formulation needs like improved adhesion, flexibility, or controlled drug release. Widely accepted by regulatory authorities, HPMC boasts established safety profiles and pharmacopeial monographs. Moreover, its availability in various grades, offering different viscosity levels, particle sizes, and substitution degrees, empowers formulators to tailor formulations to exact specifications.

The formulation includes plasticizers. Plasticizers are additives commonly used in pharmaceutical formulations to modify the properties of polymers, particularly in film coatings for tablets and capsules. These substances enhance flexibility, improve film formation, and ensure proper adhesion to the substrate. Examples include triacetin, polyethylene glycol, propylene glycol, diethyl phthalate, dibutyl phthalate, tributyl citrate, etc.

In another aspect of the embodiment, the plasticizers are polyethylene glycol and propylene glycol in a concentration of 2% v/v.

Polyethylene glycol and propylene glycol are commonly used as plasticizers in pharmaceutical formulations due to their ability to improve the flexibility, processability, and performance of polymer-based coatings and matrices. The use of polyethylene glycol and propylene glycol as plasticizers in pharmaceutical formulations offers several benefits, including flexibility enhancement, improved processability, and compatibility with various polymers. Their safety, regulatory acceptance, and multifunctional properties make them valuable components in the development of effective and stable pharmaceutical products.

The formulation includes Permeation enhancer. Permeation enhancers, also known as absorption enhancers or penetration enhancers, are substances used in pharmaceutical formulations to increase the absorption of drugs across biological barriers, such as the skin, mucous membranes, or epithelial linings. These enhancers work by various mechanisms to improve the permeability of drugs, thereby enhancing their bioavailability and therapeutic efficacy.

In another aspect of the embodiment, the Permeation enhancer is Eucalyptus oil in a concentration of 1% v/v.

Eucalyptus oil, derived from the leaves of the eucalyptus tree, is known for its medicinal properties, including its ability to act as a permeation enhancer in pharmaceutical formulations. Eucalyptus oil, rich in bioactive compounds like terpenes (e.g., 1,8-cineole) and flavonoids, has demonstrated the ability to enhance skin permeability by disrupting the stratum corneum, the outermost layer of the skin. This disruption facilitates the penetration of drugs through the skin barrier.

Moreover, eucalyptus oil's capacity to increase the solubility of hydrophobic drugs in the oil phase aids in their partitioning into the skin, enhancing absorption, particularly beneficial for poorly water-soluble drugs. Its lipophilic nature fosters interaction with lipid components of the skin, promoting drug diffusion across the skin barrier, thereby potentially leading to quicker onset of action and improved therapeutic outcomes. Additionally, incorporation of eucalyptus oil into transdermal drug delivery systems, such as patches or gels, can further optimize drug permeation, facilitating systemic absorption or localized therapy.

The formulation includes Solvent. Solvents play essential roles in pharmaceutical formulations, serving various purposes such as dissolving active pharmaceutical ingredients (APIs), facilitating drug delivery, and ensuring product stability.

In another aspect of the embodiment, the solvent is ethanol.

Ethanol, also known as ethyl alcohol, is a widely used solvent in pharmaceutical formulations due to its versatile properties and compatibility with a variety of active pharmaceutical ingredients (APIs) and excipients. Ethanol contributes to enhanced drug delivery by increasing the permeability of biological membranes, thereby improving drug absorption and bioavailability, notably in transdermal patches. Furthermore, it often serves as a cosolvent alongside other solvents to optimize drug solubility and formulation properties, ensuring compatibility with excipients and enhancing stability.

In another embodiment, the polyherbal film forming spray formulation comprising,
a) 4% v/v of mixture of extracts of Aegle Marmelos, Ocimum Basilicum and Tabernaemontana divaricata;
b) 2% w/v of Hydroxypropyl methylcellulose;
c) 2% v/v of Propylene glycol and Polyethylene glycol;
d) 1% v/v of Eucalyptus oil;
e) Ethanol
wherein the percentages are with respect to total weight of the formulation.

In another aspect of embodiment, the mixture of extracts of Aegle Marmelos, Ocimum Basilicum and Tabernaemontana divaricata are present in the weight ratio of 1:1:1.

In another aspect of embodiment, the propylene glycol and polyethylene glycol are present in the weight ratio of 1:1.

In another embodiment, the present invention relates to a process for preparation of a polyherbal film forming spray formulation comprises the steps of;
a) Dissolving hydroxypropyl methylcellulose into ethanol to form solution I;
b) Dissolving the extract of Aegle Marmelos, extract of Ocimum Basilicum, and extract of Tabernaemontana divaricata in ethanol to form a solution II;
c) Mixing the solution I and solution II to form solution III;
d) Adding propylene glycol, polyethylene glycol and Eucalyptus oil to solution III and adjusting the volume with ethanol to obtain the polyherbal film-forming spray formulation.

EXAMPLES
Example 1 - Preparation of Extracts of Aegle Marmelos, Ocimum Basilicum, and Tabernaemontana divaricata
a) Extraction process of Aegle Marmelos
• The extraction method used was Soxhlet extraction.
• 20g of coarse powder of Aegle Marmelos was exposed to 300ml ethanol as solvent at room temperature for 24 hours. Solvent recovery was carried out.
• The diluted extract collected was concentrated by employing evaporation technique using a water bath and a viscous mass was obtained.
• The crude extract was transferred to ambered coloured bottle to prevent oxidation and was used further.

b) Extraction process of Ocimum Basilicum
• 20g of Ocimum Basilicum coarse powder was exposed to 200ml of ethanol solvent at room temperature for 6 hours in a Soxhlet apparatus. After that, solvent recovery was done.
• The diluted extract collected was concentrated by employing evaporation technique using a water bath and a viscous mass was obtained.
• The crude extract was transferred to ambered coloured bottle to prevent oxidation.

c) Extraction process of Tabernaemontana divaricata
• 20g of powder of Tabernaemontana divaricata was exposed to 250ml of ethanol in a Soxhlet apparatus at room temperature for 19 hours. Solvent recovery was also carried out.
• This extract was obtained by Soxhlet method.
• The diluted extract collected was concentrated by employing evaporation technique using a water bath and a viscous mass was obtained.
• The crude extract was stored in an ambered coloured bottle to prevent oxidation and for further use.

Example 2 - Polyherbal film-forming spray formulation according to invention
SN Ingredients Quantity
1 Mixture of Extracts of AM: OB: TD
(1:1:1) 4ml (1000µg /ml)
2 Hydroxypropyl methylcellulose E 15 2 gm
3 Propylene glycol 1ml
4 Propyl ethylene glycol 1ml
5 Eucalyptus oil 1ml
6 Ethanol q. s
q.s - Quantity Sufficient; AM - Aegle Marmelos; OB - Ocimum Basilicum; TD - Tabernaemontana divaricata
Table no.1 - Polyherbal film-forming spray according to invention

Manufacturing Process of polyherbal film forming spray formulation -
1. Hydroxypropyl Methylcellulose was dissolved in the ethanol to obtain solution I;
2. Extract of Aegle Marmelos, extract of Ocimum Basilicum, and extract of Tabernaemontana divaricata were dissolved in ethanol to obtain solution II;
3. Solution I and solution II were mixed together to form solution III;
4. Propylene glycol, polyethylene glycol and Eucalyptus oil were added to the solution III;
5. Adjustment of volume was done using ethanol and the final formulation of polyherbal film forming spray was obtained.

Characterization of Polyherbal film forming spray formulation -

Physiochemical Evaluation: (Refer table No.2)
1. Physical Attributes: The polyherbal film-forming spray was stored at room temperature (30 ± 2°C) for 0, 7 and 14 days. Visual inspections were conducted to assess solution clarity, film thickness, and the appearance of white spots on the surface.

2. Evaporation Time: The film-forming spray was applied onto bagasse paper suspended from a precise balance in a fume hood. Analytical methods were used to measure the loss in weight of the bagasse paper/solvent liquid over time as the solvent evaporated.

3. Spray Volume: The spray formulations underwent quantitative testing to determine the average volume per dosage. Ten sprays were dispensed into a glass beaker, and the volume per spray was calculated using an analytical balance.

4. pH Measurement: About 20 mL of the film-forming spray solution was added to a 30 mL glass beaker. The pH values of ten spray samples were measured at intervals of 0, 7 and 14 days.

SN Test Observations
1. Physical Attributes Clear, translucent film and smooth
2. Evaporation Time 20 seconds
3. Spray Volume (ml) 0.121
4. pH Measurement 6.8
Table no. 2 - Physiochemical evaluation of the Polyherbal film forming spray.

Minimum Inhibitory Concentration (MIC): (Refer table no.3)
The study evaluated the antimicrobial activity of extract of Aegle Marmelos, extract of Ocimum Basilicum, and extract of Tabernaemontana Divaricata individually as well as in combination against various bacterial strains was performed.
E. coli: Aegle Marmelos showed the highest activity with the largest zone of inhibition at 1000 µg/ml. The combination of all three extracts (AM + OB + TD) showed the highest zone of inhibition (14mm) with an MIC of 1000 µg/ml.
Salmonella typhi: Ocimum Basilicum was most effective alone (13mm zone of inhibition at 1000 µg/ml), and the combination of all three extracts also showed a 13mm zone of inhibition at the same concentration.
S. aureus: Aegle Marmelos exhibited the highest activity alone showing a 12mm zone of inhibition, with the combination of all three extracts showing a 13mm zone of inhibition at 1000 µg/ml.
Bacillus subtilis: This strain was the least sensitive, with the combination of all three extracts showing the highest zone of inhibition (11mm) at 1000 µg/ml.
The positive control, Chloramphenicol, consistently exhibited a larger zone of inhibition than any of the extracts. The study concluded that the combination of extracts resulted in better antimicrobial activity compared to individual extracts, with the combination showing the highest effectiveness at an MIC of 1000 µg/ml for all tested bacteria.

ORGANISM MIC (µg/ml) ZONE OF INHIBITION
Extract of AM Extract of OB Extract of TD Extracts of AM+OB+TD
E. coli 1000 12mm 11mm 11mm 14mm
500 11mm 10mm 10mm 12mm
250 9mm 8mm 8mm 10mm
100 7mm 7mm 7mm 9mm
Solvent 7mm 7mm 7mm 7mm
C.P 30mm 30mm 30mm 30mm
S. typhi 1000 11mm 13mm 11mm 13mm
500 10mm 11mm 10mm 12mm
250 9mm 10mm 8mm 10mm
100 7mm 8mm 7mm 9mm
Solvent 7mm 6mm 6mm 7mm
C.P 22mm 22mm 22mm 20mm
S.aureus 1000 12mm 11mm 9mm 13mm
500 9mm 10mm 8mm 11mm
250 8mm 9mm 8mm 10mm
100 7mm 7mm 7mm 8mm
Solvent 6mm 6mm 6mm 7mm
C.P 18mm 18mm 18mm 12mm
B. subtilis 1000 8mm 11mm 9mm 11mm
500 7mm 9mm 7mm 9mm
250 7mm 7mm 6mm 8mm
100 6mm 6mm 6mm 7mm
Solvent 6mm 6mm 6mm 6mm
C.P 20mm 20mm 20mm 15mm
AM - Aegle Marmelos; OB - Ocimum Basilicum; TD - Tabernaemontana divaricata; C.P - Chloramphenicol
Table no. 3 - Minimum Inhibitory Concentration (MIC) of the herbal extracts












We Claim:
1. A Polyherbal film-forming spray formulation comprising,
a) Extract of Aegle Marmelos;
b) Extract of Ocimum Basilicum;
c) Extract of Tabernaemontana divaricata; and
d) Pharmaceutically acceptable inert excipients.

2. The formulation as claimed in claim 1, wherein the pharmaceutically acceptable inert excipients are film forming agent, plasticizers, permeation enhancer and solvent.

3. The formulation as claimed in claim 2, wherein the film-forming agent is Hydroxypropyl methylcellulose.

4. The formulation as claimed in claim 2, wherein the plasticizers are Propylene glycol and Polyethylene glycol.

5. The formulation as claimed in claim 2, wherein the permeation enhancer is Eucalyptus oil.

6. The formulation as claimed in claim 2, wherein the solvent is Ethanol.

7. A Polyherbal film-forming spray formulation comprising,
a) 4% v/v of mixture of extract of Aegle Marmelos, Ocimum Basilicum and Tabernaemontana divaricata;
b) 2% w/v of Hydroxypropyl methylcellulose;
c) 2% v/v of Propylene glycol and Polyethylene glycol;
d) 1% v/v of Eucalyptus oil;
e) Ethanol;
wherein the percentages are with respect to the total weight of the formulation.

8. The formulation as claimed in claim 7, wherein the Extract of Aegle Marmelos, Extract of Ocimum Basilicum and Extract of Tabernaemontana divaricata are present in the weight ratio of 1:1:1.

9. The formulation as claimed in claim 7, wherein the propylene glycol and polyethylene glycol are present in the weight ratio of 1:1.

10. A process for preparation of polyherbal film-forming spray formulation comprises the steps of;
a) Dissolving hydroxypropyl methylcellulose into ethanol to form solution I;
b) Dissolving the extracts of Aegle Marmelos, Ocimum Basilicum, and Tabernaemontana divaricata in ethanol to form a solution II;
c) Mixing the solution I and solution II to form solution III;
d) Adding propylene glycol, polyethylene glycol and Eucalyptus oil to solution III to obtain polyherbal film-forming spray formulation.


Dated this: 31st May 2024

Vijaykumar Shivpuje
IN/PA-1096
Agent for the Applicants

To
The Controller of Patents
The Patent Office, Mumbai




"A POLYHERBAL FILM FORMING SPRAY FORMULATION AND PREPARATION METHOD THEREOF".

ABSTRACT

The present invention relates to a polyherbal film forming spray formulation for wound healing. The polyherbal film forming spray comprises extract of Aegle Marmelos, Ocimum Basilicum and Tabernaemontana divaricata along with pharmaceutically inert excipients. The invention further provides method of preparation of a polyherbal film forming spray for wound healing.

, Claims:We Claim:
1. A Polyherbal film-forming spray formulation comprising,
a) Extract of Aegle Marmelos;
b) Extract of Ocimum Basilicum;
c) Extract of Tabernaemontana divaricata; and
d) Pharmaceutically acceptable inert excipients.

2. The formulation as claimed in claim 1, wherein the pharmaceutically acceptable inert excipients are film forming agent, plasticizers, permeation enhancer and solvent.

3. The formulation as claimed in claim 2, wherein the film-forming agent is Hydroxypropyl methylcellulose.

4. The formulation as claimed in claim 2, wherein the plasticizers are Propylene glycol and Polyethylene glycol.

5. The formulation as claimed in claim 2, wherein the permeation enhancer is Eucalyptus oil.

6. The formulation as claimed in claim 2, wherein the solvent is Ethanol.

7. A Polyherbal film-forming spray formulation comprising,
a) 4% v/v of mixture of extract of Aegle Marmelos, Ocimum Basilicum and Tabernaemontana divaricata;
b) 2% w/v of Hydroxypropyl methylcellulose;
c) 2% v/v of Propylene glycol and Polyethylene glycol;
d) 1% v/v of Eucalyptus oil;
e) Ethanol;
wherein the percentages are with respect to the total weight of the formulation.

8. The formulation as claimed in claim 7, wherein the Extract of Aegle Marmelos, Extract of Ocimum Basilicum and Extract of Tabernaemontana divaricata are present in the weight ratio of 1:1:1.

9. The formulation as claimed in claim 7, wherein the propylene glycol and polyethylene glycol are present in the weight ratio of 1:1.

10. A process for preparation of polyherbal film-forming spray formulation comprises the steps of;
a) Dissolving hydroxypropyl methylcellulose into ethanol to form solution I;
b) Dissolving the extracts of Aegle Marmelos, Ocimum Basilicum, and Tabernaemontana divaricata in ethanol to form a solution II;
c) Mixing the solution I and solution II to form solution III;
d) Adding propylene glycol, polyethylene glycol and Eucalyptus oil to solution III to obtain polyherbal film-forming spray formulation.

Documents