WO2020213002A1 - Matrice de pansement biocompatible et biodégradable, et sa préparation - Google Patents

Matrice de pansement biocompatible et biodégradable, et sa préparation Download PDF

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Publication number
WO2020213002A1
WO2020213002A1 PCT/IN2019/050640 IN2019050640W WO2020213002A1 WO 2020213002 A1 WO2020213002 A1 WO 2020213002A1 IN 2019050640 W IN2019050640 W IN 2019050640W WO 2020213002 A1 WO2020213002 A1 WO 2020213002A1
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range
mixture
bio
scaffold
active composition
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PCT/IN2019/050640
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English (en)
Inventor
Rajan Datt
Siddharth Pandey
Pallavi SHRIVASTAVA
Supriya Srivastava
Sumit SARASWAT
Anil Kumar Sharma
Ravi Kumar
Poonam Meena
Anupama KAKKAR
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Datt Lifesciences Pvt. Ltd.
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Publication of WO2020213002A1 publication Critical patent/WO2020213002A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/12—Ketones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/47—Euphorbiaceae (Spurge family), e.g. Ricinus (castorbean)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/82—Theaceae (Tea family), e.g. camellia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/906—Zingiberaceae (Ginger family)
    • A61K36/9066—Curcuma, e.g. common turmeric, East Indian arrowroot or mango ginger
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
    • A61L26/0052—Mixtures of macromolecular compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0061—Use of materials characterised by their function or physical properties
    • A61L26/0066—Medicaments; Biocides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/30—Compounds of undetermined constitution extracted from natural sources, e.g. Aloe Vera

Definitions

  • the present disclosure relates to a field of polymer chemistry.
  • biodegradable and biocompatible antimicrobial dressing for application on infected wound, chronic wound, diabetic foot ulcer, negative pressure wound therapy, bum wounds and the like.
  • wound dressings to cover and protect wounds is very well known.
  • the wound dressing should provide a sterile environment at the wound site and should rapidly absorb wound exudates while maintaining a moist wound surface.
  • the dressing should interfere as little as possible with wound healing process and should be easy to remove and replace with minimal trauma.
  • the wound dressing should be inexpensive to make, as well as should be conformable to all skin surfaces.
  • the process of wound healing consists of three phases during which the injured tissue is repaired, regenerated, and new tissue is reorganized into a scar.
  • the fibroblasts, endothelial cells, and epithelial cells migrate to the wound site. These fibroblasts produce collagen that is necessary for wound repair.
  • epithelial cells migrate from the free edges of the tissue across the wound. This event is succeeded by proliferation of epithelial cells at the periphery of the wound.
  • Researches has already shown that re-epithilialization is enhanced by the presence of occlusive wound dressings which maintain a moisture barrier.
  • a large variety of treatment and modalities are available for the treatment of wounds and ulcers. These range from applications of antibiotics, occlusive layers, bandages, poultices, mechanical devices that reduce evaporation of water and many others.
  • all of these modalities have one drawback in common; they all enhance wound healing by supporting the body mechanisms to heal the wound.
  • the passive healing process results in much disappointment because the body may have compromised immunity or other body functions that may not work optimally.
  • US 6,027,728 comprises a selection of herbal materials with curative effects combined in a powdered form for application to human skin to accomplish skin regeneration, particularly for application to human skin affected with eczema, psoriasis, allergic reactions, inflammatory rash and the like.
  • the application of the herbal powder to the skin is intended to cause a temporary inflammation.
  • US 5,766,614 is a new bum treatment composition which provides healing to the skin of people who have received bums or are afflicted with other skin complications that require healing.
  • US 6,126,950 relates to a formulation of herbal cream for cracked heels and palms. It is comprised of a natural wax as an emulsifier, extract of Curcuma and the gum of Acacia or Colophonium or Shorea.
  • the material/ therapeutic compositions should be able to improve or actively regenerate the skin, dermis and epidermis.
  • a bio-compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1 : 1 to 3 : 1.
  • a process for preparing the bio-compatible dressing matrix comprising: (i) a scaffold comprising gelatin and chitosan; and (ii) an active composition comprising a combination of curcumin, Emblica officinalis extract and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, the process comprising: (a) contacting gelatin with water, to obtain a first mixture; (b) homogenizing the first mixture, to obtain a homogenized first mixture; (c) contacting the homogenized first mixture with chitosan, to obtain a second mixture; (d) homogenizing the second mixture, to obtain a homogenized second mixture; (e) casting the homogenized second mixture, to obtain scaffold cast; (f) processing the scaffold cast, to obtain processed scaffold cast; (g) obtaining the active composition comprising curcumin, Emblica officinalis extract and Camellia
  • a method of treating a wound comprising, contacting the bio-compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, with the wound.
  • Figure 1 illustrates the schematic representation of the serial dilution of the bacterial species, in accordance with an implementation of the present disclosure.
  • Figure 2 shows the layout of the experiment done to observe growth of the bacterial species after treatment with extracts, positive control, and negative control, in accordance with an implementation of the present disclosure.
  • Figure 3 shows the pictorial representation of the migration of Fibroblast cells after treatment with the active composition of the present disclosure, in accordance with an implementation of the present disclosure.
  • Figure 4 shows the phase contrast microscopic pictures of the migration of Keratinocyte cells after treatment with the active composition of the present disclosure, in accordance with an implementation of the present disclosure.
  • Figure 5 depicts the antimicrobial activity of the bio-compatible dressing matrix against E. coli, in accordance with an implementation of the present disclosure.
  • Figure 6 depicts the wound healing study images of Sprague Dawley Rat as on day 1 and day 21, in accordance with an implementation of the present disclosure.
  • Figure 7 depicts the histological images of wound healing activity of the bio-compatible dressing matrix of the present disclosure, in accordance with an implementation of the present disclosure.
  • the articles“a”,“an” and“the” are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.
  • Ratios, concentrations, amounts, and other numerical data may be presented herein in a range format. It is to be understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited.
  • a temperature range of about 150-170°C should be interpreted to include not only the explicitly recited limits of about 150°C to about 170°C, but also to include sub-ranges, such as 150-165°C, 160-170°C, and so forth, as well as individual amounts, including fractional amounts, within the specified ranges, such as 150.2 °C, and 165.5 °C, for example.
  • biocompatible means that said dressing matrix is non-toxic and do not cause irritation to the tissues in the vicinity, to an extent that the medical professional finds it safe to use said dressing matrix on the patient.
  • a“dressing matrix exhibits effective antimicrobial property” refers to the amount needed in order to kill bacteria in a patient to a non-detectable degree.
  • the actual amount that is determined to be an effective will vary depending on factors such as the size, condition, sex and age of the patient and can be more readily determined by the care giver.
  • a bio-compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1.
  • the critical aspect of the bio-compatible dressing matrix of the present disclosure is the sequential timed patterned physico chemical treatment of the two polymers (gelatin and chitosan) by using a very simplified process so as to obtain a stable molecular interaction and orientation between the molecules of the two polymers to get highly stabilized porous scaffold without using any cross-linker or any integrated harmful chemicals.
  • the process for preparing the bio-compatible dressing matrix comprises the breakdown of stabilized dried air bubbles and the preparation of Polyelectrolyte complex (PEC), which is achieved using a specifically designed aspect ratio of a system for agitation/homogenization. Therefore, the present disclosure provides a ready-to-use biodegradable and biocompatible matrix and a process for preparing the same.
  • the matrix of the present disclosure comprises natural porous scaffold micro-matrix based structure mainly of Polyelectrolyte complex (PEC) acting as carrier of plurality of therapeutics.
  • the matrix has differential porosity and regulated pores with interconnected small voids on the same platform in a molecularly integrated matrix, such that said matrix acts as an active composition (drug) carrier and transporter to supply and transfer said therapeutics and pharmaceuticals into wounds, whenever applied on the wound.
  • PEC Polyelectrolyte complex
  • the bio-compatible dressing matrix of the present disclosure with its specific composition actively promotes healing by stimulating stem cells that are present even in deep wounds and burns to regenerate the lost tissue.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio of 2: 1.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1: 1 to 3: 1 , wherein the active composition has an amount in a range of lOOpg/cm 2 - 20mg/cm 2 with respect to the bio-compatible dressing matrix.
  • the active composition has an amount in a range of lmg/cm 2 - 15mg/cm 2 with respect to the bio-compatible dressing matrix. In yet another embodiment of the present disclosure, the active composition has an amount in a range of 3mg/cm 2 - 10mg/cm 2 with respect to the bio-compatible dressing matrix.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1: 1 to 3: 1, wherein the active composition has an amount in a range of 0.8 mg/cm 2 - 1.4 mg/cm 2 with respect to the bio-compatible dressing matrix.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio of 2: 1, wherein the active composition has an amount in a range of lOOpg/cm 2 - 20mg/cm 2 with respect to the bio-compatible dressing matrix.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, wherein the active composition comprises Emblica officinalis extract having a weight percentage in a range of 0.5-20% with respect to the active composition, Camellia sinensis extract having a weight percentage in a range of 1-20% with respect to the active composition, curcumin having a weight percentage in a range of 0.1-12% with respect to the active composition.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1: 1 to 3:1, wherein the active composition comprises Emblica officinalis extract having a weight percentage in a range of 5-15% with respect to the active composition, Camellia sinensis extract having a weight percentage in a range of 5-15% with respect to the active composition, curcumin having a weight percentage in a range of 5-10% with respect to the active composition.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, wherein the active composition comprises Emblica officinalis extract having a weight percentage in a range of 5-10% with respect to the active composition, Camellia sinensis extract having a weight percentage in a range of 5-10% with respect to the active composition, curcumin having a weight percentage in a range of 5-8.5% with respect to the active composition.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, wherein the active composition comprises Emblica officinalis extract having a weight percentage is 7 % with respect to the active composition, Camellia sinensis extract having a weight percentage is 7 % with respect to the active composition, curcumin having a weight percentage is 7 % with respect to the active composition.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio of 2:1, wherein the active composition comprises Emblica officinalis extract having a weight percentage is 7 % with respect to the active composition, Camellia sinensis extract having a weight percentage is 7 % with respect to the active composition, curcumin having a weight percentage is 7 % with respect to the active composition.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio of 2:1, wherein the active composition comprises Emblica officinalis extract having a weight percentage in a range of 0.5-20% with respect to the active composition, Camellia sinensis extract having a weight percentage in a range of 1-20% with respect to the active composition, curcumin having a weight percentage in a range of 0.1-12% with respect to the active composition.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, wherein gelatin has a weight percentage in a range of 1-4% with respect to the matrix, and chitosan has a weight percentage in a range of 0.5-2.5% with respect to the matrix.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, wherein gelatin has a weight percentage in a range of 1-4% with respect to the matrix, and chitosan has a weight percentage in a range of 0.5-2.5% with respect to the matrix, and wherein the active composition comprises Emblica officinalis extract having a weight percentage in a range of 0.5-20% with respect to the active composition, Camellia sinensis extract having a weight percentage in a range of 1-20% with respect to the active composition, curcumin having a weight percentage in a range of 0.1-12% with respect to the active composition.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, wherein gelatin has a weight percentage in a range of 1-4% with respect to the matrix, and chitosan has a weight percentage in a range of 0.5-2.5% with respect to the matrix, and wherein the active composition comprises Emblica officinalis extract having a weight percentage in a range of 5-10% with respect to the active composition, Camellia sinensis extract having a weight percentage in a range of 5-10% with respect to the active composition, curcumin having a weight percentage in a range of 5-8.5% with respect to the active composition.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio of 2:1, wherein gelatin has a weight percentage in a range of 1-4% with respect to the matrix, and chitosan has a weight percentage in a range of 0.5-2.5% with respect to the matrix.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio of 2:1, wherein gelatin has a weight percentage in a range of 1-4% with respect to the matrix, and has a bloom strength in a range of 50-300, and chitosan has a weight percentage in a range of 0.5-2.5% with respect to the matrix and has degree of acetylation in a range of 75- 95%.
  • a process for preparing the bio-compatible dressing matrix comprising: (i) a scaffold comprising gelatin and chitosan; and (ii) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, said process comprising: (a) contacting gelatin with water, to obtain a first mixture; (b) homogenizing the first mixture, to obtain a homogenized first mixture; (c) contacting the homogenized first mixture with chitosan, to obtain a second mixture; (d) homogenizing the second mixture, to obtain a homogenized second mixture; (e) casting the homogenized second mixture, to obtain scaffold cast; (f) processing the scaffold cast, to obtain processed scaffold cast; (g) obtaining the active composition comprising curcumin, Emblica officinalis extract and Camellia
  • a process for preparing the bio-compatible dressing matrix comprising: (i) a scaffold comprising gelatin and chitosan; and (ii) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, said process comprising: (a) contacting gelatin with water is done at a temperature in a range of 55-65°C, to obtain a first mixture; (b) homogenizing the first mixture, to obtain a homogenized first mixture; (c) contacting the homogenized first mixture with chitosan, to obtain a second mixture; (d) homogenizing the second mixture, to obtain a homogenized second mixture; (e) casting the homogenized second mixture, to obtain scaffold cast; (f) processing the scaffold cast, to obtain processed scaffold cast; (g) obtaining the active composition comprising cur
  • a process for preparing the bio-compatible dressing matrix comprising: (i) a scaffold comprising gelatin and chitosan; and (ii) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, said process comprising: (a) contacting gelatin with water is done at a temperature in a range of 55-65°C, to obtain a first mixture; (b) homogenizing the first mixture is done using a homogenizer in a range of 2000-3000 rpm for a time period in a range of 15-25 minutes, to obtain a homogenized first mixture; (c) contacting the homogenized first mixture with chitosan, to obtain a second mixture; (d) homogenizing the second mixture, to obtain a homogenized second mixture; (e) casting the homogenized second
  • a process for preparing the bio-compatible dressing matrix comprising: (i) a scaffold comprising gelatin and chitosan; and (ii) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, said process comprising: (a) contacting gelatin with water is done at a temperature in a range of 55-65°C, to obtain a first mixture; (b) homogenizing the first mixture is done using a homogenizer in a range of 2000-3000 rpm for a time period in a range of 15-25 minutes, to obtain a homogenized first mixture; (c) contacting the homogenized first mixture with chitosan, to obtain a second mixture; (d) homogenizing the second mixture, to obtain a homogenized second mixture; (e) casting the homogenized second
  • a process for preparing the bio-compatible dressing matrix comprising: (i) a scaffold comprising gelatin and chitosan; and (ii) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, said process comprising: (a) contacting gelatin with water is done at a temperature in a range of 55-65°C, to obtain a first mixture; (b) homogenizing the first mixture is done using a homogenizer in a range of 2000-3000 rpm for a time period in a range of 15-25 minutes, to obtain a homogenized first mixture; (c) contacting the homogenized first mixture with chitosan in a range of 10-15 grams of chitosan per minute, to obtain a second mixture; (d) homogenizing the second mixture, to obtain a
  • a process for preparing the bio-compatible dressing matrix comprising: (i) a scaffold comprising gelatin and chitosan; and (ii) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, said process comprising: (a) contacting gelatin with water is done at a temperature in a range of 57-62°C, to obtain a first mixture; (b) homogenizing the first mixture is done using a homogenizer in a range of 2200-2800 rpm for a time period in a range of 17-22 minutes, to obtain a homogenized first mixture; (c) contacting the homogenized first mixture with chitosan in a range of 11-14 grams of chitosan per minute, to obtain a second mixture; (d) homogenizing the second mixture, to
  • a process for preparing the bio-compatible dressing matrix comprising: (i) a scaffold comprising gelatin and chitosan; and (ii) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, said process comprising: (a) contacting gelatin with water is done at a temperature in a range of 55-65°C, to obtain a first mixture; (b) homogenizing the first mixture is done using a homogenizer in a range of 2000-3000 rpm for a time period in a range of 15-25 minutes, to obtain a homogenized first mixture; (c) contacting the homogenized first mixture with chitosan in a range of 10-15 grams of chitosan per minute, to obtain a second mixture; (d) homogenizing the second mixture, to obtain a
  • a process for preparing the bio-compatible dressing matrix comprising: (i) a scaffold comprising gelatin and chitosan; and (ii) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, said process comprising: (a) contacting gelatin with water is done at a temperature in a range of 55-65°C, to obtain a first mixture; (b) homogenizing the first mixture is done using a homogenizer in a range of 2000-3000 rpm for a time period in a range of 15-25 minutes, to obtain a homogenized first mixture; (c) contacting the homogenized first mixture with chitosan in a range of 10-15 grams of chitosan per minute, to obtain a second mixture; (d) homogenizing the second mixture involves processing the second mixture
  • a process for preparing the bio-compatible dressing matrix comprising: (1) a scaffold comprising gelatin and chitosan; and (2) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, said process comprising: (a) contacting gelatin with water is done at a temperature in a range of 55-65°C, to obtain a first mixture; (b) homogenizing the first mixture is done using a homogenizer in a range of 2000-3000 rpm for a time period in a range of 15-25 minutes, to obtain a homogenized first mixture; (c) contacting the homogenized first mixture with chitosan is done in a range of 10-15 grams of chitosan per minute, to obtain a second mixture; (d) homogenizing the second mixture involves processing the second mixture also, to
  • a process for preparing the bio-compatible dressing matrix comprising: (1) a scaffold comprising gelatin and chitosan; and (2) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, said process comprising: (a) contacting gelatin with water, to obtain a first mixture; (b) homogenizing the first mixture, to obtain a homogenized first mixture; (c) contacting the homogenized first mixture with chitosan, to obtain a second mixture; (d) homogenizing the second mixture, to obtain a homogenized second mixture; (e) casting the homogenized second mixture, to obtain scaffold cast; (f) processing the scaffold cast, to obtain processed scaffold cast; (g) obtaining the active composition comprising curcumin, Emblica officinalis extract and Camellia sinensis extract
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, wherein the dressing matrix exhibits release of actives in a range of 60 pg/ml to 300 pg/ml in 24 hrs.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, and wherein the dressing matrix exhibits antimicrobial property.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, for use in wound healing.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, for use as antiseptic dressing.
  • a bio compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1:1 to 3:1, wherein the bio-compatible dressing matrix is used to deliver curcumin, Emblica officinalis extract and Camellia sinensis extract towards healing of infected wound, burn wound and as foam matrix for negative pressure wound therapy.
  • a method of treating a wound comprising: contacting the bio-compatible dressing matrix comprising: (a) a scaffold comprising gelatin and chitosan; and (b) an active composition comprising a combination of curcumin, Emblica officinalis extract, and Camellia sinensis extract, wherein the scaffold comprises gelatin and chitosan in a weight ratio in a range of 1 : 1 to 3 : 1 with the wound.
  • Bacterial growth media and agar were supplied by Hi-media. [00072] Studies on animal subjects (Sprague Dawley rats) were performed according to the protocols approved by IAEC (registered with CPCSEA, registration no. 1974/PO/Rc/).
  • non-aqueous extract of the fruit of Emblica officinalis, non- aqueous extract of the leaves of Camellia sinensis and Curcuma were commercially procured.
  • the non-aqueous extract of the actives can be prepared using the following process.
  • the non-aqueous solvent used in commercially procured extracts were selected from ethanol, methanol, hexane, toluene, chloroform, dichloromethane, methylene chloride, acetone. The ratio between the solvents is in the range of 1:1 - 1:3 volume/volume.
  • the respective supernatants were allowed to dry in oven at a temperature in the range of at 60°C - 65°C to obtain the respective dried supernatant mixtures.
  • the desired concentration of the non- aqueous extract of Emblica officinalis, and the non-aqueous extract of Camellia sinensis was prepared by dissolving the respective dried supernatant mixture in a 10%-20% ethanol solution followed by sonication at 5-85 amplitude for a period in the range of 15 minutes to 300 minutes. Subsequently the excess of ethanol was evaporated to obtain non-aqueous extract of Emblica officinalis, and the non- aqueous extract of Camellia sinensis respectively.
  • non-aqueous extracts procured commercially or prepared from the process described above were subjected to physico-chemical treatment by altering the pH of the solution (from basic to acidic, i.e., 12 to 3.5) followed by ultrasonic treatment at 5-25 amplitude for a period of 2 hours to 5 hours to get water soluble nano-sized fraction of non-aqueous (e.g. ethanolic) extracts, i.e., Emblica officinalis extract and Camellia sinensis extract.
  • non-aqueous extracts i.e., Emblica officinalis extract and Camellia sinensis extract.
  • Curcumin 1M solution of NaOH was prepared (pH of the solution was 12.5-12.6). 5-10% Curcumin solution was prepared in 1M solution of NaOH with continuous mixing. The prepared solution was then neutralized with 5N HC1 and then adjusted to pH 5.5 The prepared solution was then sonicated for 5 hrs with amplitude 25 and temperature maintained below 30°C.
  • the flasks were then placed in UV and 70% ethanol sterilized laminar air flow hood. 15-20 bacteriological sterilized petri plates were placed in the hood. 20 ml of the nutrient agar solution was poured in petri plates with a sterilized 50ml glass beaker. Uniform distribution of agar solution was carefully done. Further the plates were left open for 1 hour to solidify the agar solution. Once the agar solution solidified, the lids were closed, and the plates were wrapped with parafilm. These plates were then stored at 4°C until use.
  • Sub-culturing of bacterial cultures i.e., MTCC 739 and MTCC 737: 4 fresh soyabean culture medium flasks were taken out from 4°C and were kept outside to thaw to room temperature. 20 pi of bacterial culture was taken from each bacterial strain and the fresh medium flasks were inoculated respectively, for subculture. The freshly sub-cultured flasks were kept in incubator shaker to grow overnight at 121rpm and 37°C.
  • Betadine dilution 10% solution of betadine was diluted in a ratio of 1:1 in LB media i.e, to 1ml of betadine, 1ml of LB media was added so as to prepare the positive control.
  • LB media negative control
  • betadine positive control
  • Nutrient agar plates Before starting the spreading work, the required numbers of agar plates were taken out from 4°C and were kept outside to thaw to room temperature. After completion of each time point, the 96 well plate were taken out from shaker incubator and the solution was properly mixed. IOOmI from each condition was taken and dropped in the centre of agar plates. With the help of sterile spreader, the solution was uniformly spread on agar plates. The agar plates were sealed with parafilm and were overnight kept at 37° in incubator. Next day the plates were observed, and the number of colonies were written down for each plate. The results obtained are depicted in Table 2 below, and the layout of the plate is depicted in Figure 2.
  • the active composition of the present disclosure was obtained by contacting 7 % extract of Emblica officinalis, 1 % extract of Camellia sinensis extract, and 7 % of curcumin with water, to obtain a first active mixture.
  • Water used was ultra-pure water with a resistivity of 18.2 M Wah 1 .
  • the first active mixture was sonicated for a time period in a range of 1-4 hours, at amplitude in a range of 15-30 seconds at a temperature in a range of 25-35°C, to obtain a processed second active mixture.
  • the second active mixture was then lyophilized to obtain a lyophilized mixture.
  • time killed assay was done. The protocol was followed as per previously published reports [(American society for testing and materials (ASTM) Standard guide for assessment of microbial activity using a time kill procedure, E2315-03 (Reapproved 2008); Food and drug administration. Tentative final monograph for healthcare antiseptic drug products: proposed rule. Code of federal regulations, 21 CFR parts 333and 369.
  • the active composition was effective against superbugs. The same was proved by the time killed assay, wherein the population of microorganism was reduced by 99.99 % within 30 minutes of the exposure.
  • the active composition of the present disclosure comprising curcumin, Emblica officinalis extract and Camellia sinensis extract as exemplified in the Example 3 can be considered to be a promising candidate to be used for effective wound healing.
  • Fibroblasts and keratinocytes were seeded uniformly in 24 well plates at a density of 40,000-60,000 cells per well in 20% MEM. Cells were grown up to 100% culture confluency to form a monolayer. Once confluent the cells were deprived of serum in the basal MEM medium for 16-17 hrs or overnight, next day, a scratch was created on the cells monolayer using a IOOmI pipette tip. Cells were then washed with IX PBS to remove any cell debris. Once scratch was created in all wells, test samples were added in all the wells including wells for positive and negative control. Pictures were taken at regular interval. A sequence of representative images from a wound healing assay carried out on a confluent monolayer is shown in Figures 3 and 4 for fibroblasts and keratinocytes, respectively.
  • the most common information derived from the wound healing assay is the rate of gap closure, which is a measure of the speed of the collective motion of the cells.
  • the speed of wound closure and cell migration can be quantified by taking snapshot pictures with an inverted microscope at different time intervals.
  • the migration assay on human fibroblast cells and keratinocytes cells was performed by treating with three different concentration (O.lpg/ml, 0.5pg/ml and 1 pg/ml) of active composition of the present disclosure and snapshots were taken at different time intervals (0 hrs, 12 hrs, 24 hrs and 96 hrs).
  • cell migration initiated at 12 hrs in all the three concentrations (O.lpg/ml, 0.5 pg/ml and 1 pg/ml) and complete cell migration was observed after 24 hrs in O.lpg/ml.
  • the objective of this example was to evaluate the antimicrobial activity of active composition of the present disclosure as provided in Example 3, using a time kill study against bacterial strains MTCC 1687 (. Escherichia coli ) and MTCC AA ⁇ Bacillus subtilis).
  • test item along with bacterial culture of Escherichia coli and Bacillus subtilis alone and with bacterial dilutions of 10 3 and 10 6 were mixed together and incubated for different time points, i.e., 30min, 60min, 90min and 120 min in a 96 well plate and kept in incubator shaker at 37 °C and lOOrpm.
  • the active composition of the present disclosure effectively killed the challenged microorganism of both strains at pure culture and at 2 different dilutions also. Results obtained showed that killing start at 30 minutes duration and lasted after a span of 120 minutes as well. No colonies were observed on the plates till 7 days.
  • the polymers were weighed in a ratio of 2:1.
  • Gelatin solution was prepared by contacting gelatin in ultra-pure water followed by heating at 60°C to obtain a first mixture.
  • the first mixture was then homogenized using homogenizer between 2000 rpm to 3000 rpm for 10-15 min. Further, acetic acid was added so that its final concentration is 1% of the first mixture.
  • the reactants were then allowed to mix properly by continuous homogenization using the stirrer for 2 mins to obtain a homogenized first mixture.
  • Chitosan was added to the homogenized first mixture at the rate of 15 gm per minute to obtain a second mixture.
  • the second mixture was then homogenized using stirrer for 90-110 min, followed by keeping the same in an air dryer and allowing it to dry at 5°C - 18 °C and humidity 30% - 50% obtain a porous matrix (homogenized second mixture).
  • the casting tray were made as per the desired size and the homogenized mixture were casted into the casting tray to obtain a scaffold cast.
  • the desired size is a variable which depends on the size of the sample and batch size.
  • the scaffold cast was then processed further, by air drying followed by compressing it in compressing machine.
  • the scaffold was then cut into desired shapes. These scaffolds were then dipped into 6.25 % of ammonia solution for 2 hours followed by washing them with ultra-pure water 6-7 times so that the ammonia is removed from it.
  • the scaffold was then pat-dried by compressing it to remove the excess of the water to obtain the processed scaffold cast.
  • the active composition of the example 3 was then contacted with the processed scaffold cast until the active composition was completely absorbed by the processed scaffold to obtain the loaded scaffold.
  • the loaded scaffold was then intermittently freeze-dried by freezing at -40°C for 2hrs and then freeze drying at 10°C for 60 hrs at vacuum of 100 mTorr to obtain dried scaffold.
  • the dried scaffold was then kept in a conditioning chamber at 40°C and 75% Relative humidity for 15-30 min to obtain a bio-compatible dressing matrix with 10-15% of moisture in it.
  • a bio-compatible dressing matrix of the present disclosure is
  • a bio-compatible dressing matrix is a porous antimicrobial dressing composed of scaffold made from polymeric blend of biocompatible and biodegradable biopolymers loaded with the active composition of the present disclosure.
  • Various combination for the ratio of the polymers were tried to make the bio-compatible dressing matrix. It was observed that the ratio of gelatin to chitosan pertaining to 1:2 could not meet the desired parameters as they showed no antimicrobial activity or very less antimicrobial activity.
  • matrix with gelatin to chitosan weight ratio 2:1 displayed desirable physical properties of porosity, water uptake, and antimicrobial activity. The antimicrobial activity is depicted in in Figure 5, wherein zone of inhibitions are visible.
  • the bio-compatible dressing matrix of the present disclosure is effective against wound healing activity. This has been inferred based on the experiments done on burn wound in Sprague Dawley rats.
  • An insulated stainless- steel rod weighing approximately 21 g, with attached cylindrical plate of 2 ⁇ 0.2 cm diameters was used for creation of burn wound on depilated cervicothoracic skin region.
  • the plate was immersed in a 100 ⁇ 5°C boiling water bath for 5 minutes. Using the thumb and index finger of both hands, portion of skin on cervicothoracic region was pulled up from the underlying viscera and directing the two sides upwards and outwards. The heated plate was held in contact with the skin perpendicularly without applying pressure for 15 seconds to create a burn wound.
  • the present disclosure thus provides a ready-to-use biocompatible dressing matrix comprising active composition.
  • the active composition comprises curcumin, and non-aqueous extract of Emblica officinalis and Camellia sinensis.
  • the matrix comprises gelatin-chitosan polyelectrolyte complex.
  • the biocompatible dressing matrix of the present disclosure can be used in the form of a bandage dressing, porous scaffold or foam with enhanced bioavailability. Also, through the biocompatible dressing matrix of the present disclosure herbal extracts can be directly delivered to the infected site, consequently accelerating the healing effect.
  • ADVANTAGES ADVANTAGES:
  • the disclosed biocompatible dressing matrix is mainly for medical purpose and the broad range applications of the matrix can be (a) anti-microbial; (b) negative pressure wound therapy; (c) anti-inflammatory; (d) analgesic; (e) antioxidant; (g) radiation poisoning; (h) modulating immunity.
  • the disclosed biocompatible dressing matrix were also found to be effective in loading and releasing bioactive molecules.

Abstract

La présente invention concerne une matrice de pansement biocompatible comprenant : (a) un échafaudage comprenant de la gélatine et du chitosane ; et (b) une composition active comprenant une combinaison d'extrait de curcumine, d'extrait d'Emblica officinalis et d'extrait de Camellia sinensis, l'échafaudage comprenant de la gélatine et du chitosane dans un rapport pondéral compris entre 1:1 à 3:1. L'invention se rapporte en outre à un procédé de préparation de celle-ci. De plus, l'invention concerne également un procédé de traitement d'une plaie comprenant la mise en contact de la matrice de pansement biocompatible de la présente invention avec la plaie.
PCT/IN2019/050640 2019-04-17 2019-09-05 Matrice de pansement biocompatible et biodégradable, et sa préparation WO2020213002A1 (fr)

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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7750063B2 (en) * 2001-10-24 2010-07-06 Pentron Clinical Technologies, Llc Dental filling material
WO2014152959A1 (fr) * 2013-03-14 2014-09-25 Forsight Vision4, Inc. Systèmes pour l'administration intra-oculaire entretenue de composés à faible solubilité provenant d'un implant de système de pose d'orifice
US20160143786A1 (en) * 2013-06-28 2016-05-26 3M Innovative Properties Company Fibrin-coated wound dressing
US20160339140A1 (en) * 2014-03-24 2016-11-24 Datt Mediproducts Limited A Ready To Use Biodegradable And Biocompatible Device And A Method Of Preparation Thereof
US20170224867A1 (en) * 2016-02-08 2017-08-10 Datt Mediproducts Limited Ready-to-use, hydrophilic, self-dispersive, fragmentable and biodegradable porous sponge matrix and a method of manufacturing thereof
US10149924B1 (en) * 2016-11-28 2018-12-11 Datt Mediproducts Limited Ready to use biodegradable and biocompatible artificial skin substitute and a method of preparation thereof
US20180369315A1 (en) * 2016-08-09 2018-12-27 Datt Mediproducts Limited Multifunctional formulation comprised of natural ingredients and method of preparation/manufacturing thereof

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7750063B2 (en) * 2001-10-24 2010-07-06 Pentron Clinical Technologies, Llc Dental filling material
WO2014152959A1 (fr) * 2013-03-14 2014-09-25 Forsight Vision4, Inc. Systèmes pour l'administration intra-oculaire entretenue de composés à faible solubilité provenant d'un implant de système de pose d'orifice
US20160143786A1 (en) * 2013-06-28 2016-05-26 3M Innovative Properties Company Fibrin-coated wound dressing
US20160339140A1 (en) * 2014-03-24 2016-11-24 Datt Mediproducts Limited A Ready To Use Biodegradable And Biocompatible Device And A Method Of Preparation Thereof
US20170224867A1 (en) * 2016-02-08 2017-08-10 Datt Mediproducts Limited Ready-to-use, hydrophilic, self-dispersive, fragmentable and biodegradable porous sponge matrix and a method of manufacturing thereof
US20180369315A1 (en) * 2016-08-09 2018-12-27 Datt Mediproducts Limited Multifunctional formulation comprised of natural ingredients and method of preparation/manufacturing thereof
US10149924B1 (en) * 2016-11-28 2018-12-11 Datt Mediproducts Limited Ready to use biodegradable and biocompatible artificial skin substitute and a method of preparation thereof

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