RU2818431C2 - Pharmaceutical compositions for delivery and use thereof - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: present invention relates to compositions and methods for repair and regeneration of skin and mucous membranes. According to some embodiments, the compositions described by this disclosure contain natural products (for example, extracts) in combination with biotechnological auxiliary systems, such as biological surfactants, suitable for repair of injured skin, such as mucous membrane (for example, oral mucosa).

EFFECT: restoration of damaged skin.

22 cl, 6 dwg, 9 tbl, 7 ex

Description

Related applications

[001] This application claims priority under 35 U.S.C. 119(e) as filed on U.S. Provisional Application No. 62/791005, filed January 10, 2019, entitled “COMPOSITIONS FOR DRUG DELIVERY AND USE,” the entire contents of which are incorporated herein by reference.

BACKGROUND OF THE INVENTION

[002] Methods for treating damaged tissue, such as skin or mucous membranes, typically involve topical or systemic application of therapeutic agents (eg, antibacterial drugs) in combination with wound dressing and mechanical or enzymatic debridement. While these treatments are usually sufficient to speed up the healing of minor cuts and infections, more severely damaged tissue such as chronic wounds, infected skin, etc. often cannot be properly treated with these methods. The main challenge for existing methods is to create a tissue microenvironment that is both impervious to microbial growth and creates molecular conditions that promote tissue regeneration and healing.

Brief Disclosure of the Present Invention

[003] Aspects of the present invention relate to compositions and methods useful for treating damaged skin. The present invention is based in part on drug delivery compositions (systems) containing one or more of the following components: a surfactant (eg, a surfactant), a hydrating agent (eg, a hygroscopic agent), and a carbonate buffer that maintains the pH of the composition in the range from approximately pH 7.5 to pH 9.5. Without wishing to be bound by any particular theory, when applied to damaged tissue (e.g., skin, mucous membrane, etc.) of a subject (e.g., a human subject), the compositions described herein create a microenvironment that promotes tissue healing and regeneration, simultaneously providing 1) molecular sanitation of the wound; 2) proper hydration; and 3) anti-inflammatory and antimicrobial effects (eg, removal of bacterial biofilm as a result of disruption of the structure of the exopolymeric polysaccharide framework). In accordance with some embodiments, the properties and/or activities of the compositions described herein are unexpected from the perspective of currently available wound healing compositions, which typically have pH values ranging from neutral (e.g., approximately pH 7.0) to acidic (eg approximately pH 5.0-6.0).

[004] Accordingly, in some aspects, the present invention provides a composition comprising a drug delivery system comprising a carbonate buffer solution, at least two different biological surfactants, at least one hygroscopic agent, and at least one antioxidant ; and at least one biologically active agent, the composition having a pH in the range from about 7.5 to about 9.5.

[005] In accordance with some embodiments, the carbonate buffer solution contains one of the following ions: sodium, potassium, calcium, and magnesium. In accordance with some embodiments, the carbonate buffer solution is a sodium bicarbonate buffer solution.

[006] In accordance with some embodiments, the biological surfactants of the composition are selected from a glycolipid, a lipopeptide, and a polymeric biological surfactant.

[007] In accordance with some embodiments, the glycolipid is selected from rhamnolipid, sophorolipid, trehalolipid, cellobiolipid, mannosylerythritol lipid, and any combination thereof. In accordance with some embodiments, the biological surfactants contain at least one rhamnolipid and at least one sophorolipid.

[008] In accordance with some embodiments, the lipopeptide is selected from surfactin, plipastatin, bacillomycin, fengycin, subtilisin, gramicidin, polymyxin, and any combination thereof.

[009] In accordance with some embodiments, the polymeric biological surfactant is selected from emulsan, biodispersan, liposan, mannan-lipid-protein complex, carbohydrate-lipid-protein complex, and any combination thereof.

[010] In accordance with some embodiments, the total amount of biological surfactants in the composition ranges from about 1.5% (w/w) to about 10% (w/w). In some embodiments, the ratio of rhamnolipid to sophorolipid in the composition ranges from about 1:9 (e.g., 1% (w/w) to 99% (w/w)) to about 9:1 (eg, from 99% (w/w) to about 1% (w/w)). In some embodiments, the ratio is approximately 50% to 50% (eg, 1:1).

[011] In accordance with some embodiments, the hygroscopic agent includes an agent selected from a glycol polymer, a glycosaminoglycan, and a cellulose polymer. In some embodiments, the glycol polymer includes a polyethylene glycol polymer, optionally the polyethylene glycol polymer containing from 2 polymer subunits to about 50,000 polymer subunits. In accordance with some embodiments, the glycosaminoglycan is hyaluronic acid. In some embodiments, the cellulosic polymer includes carboxymethylcellulose or hydroxyethylcellulose. In accordance with some embodiments, the amount of hygroscopic agent in the composition ranges from about 1% (w/w) to about 10% (w/w).

[012] In accordance with some embodiments, at least one antioxidant in the composition is a lipophilic antioxidant. In some embodiments, the lipophilic antioxidant is butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), and/or tocopherol. In accordance with some embodiments, the tocopherol is alpha-tocopherol.

[013] In accordance with some embodiments, the composition contains at least one biologically active agent. In accordance with some embodiments, the biologically active agent is a small molecule, protein, nucleic acid, or biologically active extract.

[014] In accordance with some embodiments, the biologically active extract is obtained (eg, extracted) from one or more types of propolis (eg, propolis mixture). In some embodiments, the propolis (eg, propolis mixture) includes green propolis, brown propolis, red propolis, or a combination thereof (eg, green and brown propolis, red, green and brown propolis, etc.). In accordance with some embodiments, the natural bioactive extract (eg, propolis extract) contains one or more of the following: a flavonoid, artepillin C, a triterpenoid, an isoflavonoid, and an aromatic acid. In accordance with some embodiments, the ratio of green propolis to brown propolis in the composition ranges from about 1:9 to about 9:1. In accordance with some embodiments, the total amount of biologically active propolis extract in the composition ranges from about 5% (w/w) to about 20% (w/w).

[015] In accordance with some embodiments, the biologically active natural extract is obtained (eg, extracted) from one or more species of seaweed (eg, a mixture of seaweed). In some embodiments, the one or more seaweed species is selected from the genus Enteromorpha, Ulva, Monostroma, Codium, Caulerpa, Bryopsis, Porphyra, and Laminaria. In accordance with some embodiments, the biologically active natural extract (eg, seaweed extract) contains one or more sulfated polysaccharides. In accordance with some embodiments, the one or more sulfated polysaccharides are selected from carrageenan, galactan, ulvan, and fucoidan. In accordance with some embodiments, the biologically active extract further comprises Aloe vera extract. In accordance with some embodiments, the total amount of bioactive seaweed extract in the composition ranges from about 5% (w/w) to about 20% (w/w).

[016] In accordance with some embodiments, the biologically active natural extract contains one or more carotenoids and/or vitamin A (or a derivative thereof). In accordance with some embodiments, the bioactive extract containing carotenoids is obtained (eg, extracted) from carrot oil. In some embodiments, the bioactive natural carrot oil extract contains one or more B vitamin complexes. In some embodiments, the one or more B vitamin complexes are a pantothenic acid derivative or a derivative or analog thereof. In accordance with some embodiments, the vitamin B complex includes dexpanthenol. In accordance with some embodiments, the total amount of biologically active natural carrot oil extract in the composition ranges from about 1% (w/w) to about 5% (w/w).

[017] In accordance with some embodiments, the composition further comprises one or more proteins (eg, animal proteins, extracellular matrix proteins, etc.). In some embodiments, the protein includes collagen, albumin, or a combination thereof.

[018] In accordance with some embodiments, the biologically active natural extract is obtained (eg, extracted) from a plant. In accordance with some embodiments, the biologically active natural extract is obtained (eg, extracted) from Melissa officinalis. In some embodiments, the bioactive natural extract of Melissa officinalis contains one or more B vitamin complexes. In some embodiments, the one or more B vitamin complexes contain methylcobalamin and/or cyanocobalamin. In some embodiments, the one or more B vitamin complexes are a pantothenic acid derivative or a derivative or analog thereof, optionally wherein the B vitamin complex comprises dexpanthenol. In accordance with some embodiments, the total amount of biologically active natural extract of Melissa officinalis in the composition ranges from about 1% (w/w) to about 5% (w/w).

[019] In accordance with some embodiments, the composition described herein is formulated as a solid (eg, a powder, such as a lyophilized powder), liquid, gel (eg, hydrogel), or foam. In accordance with some embodiments, the liquid is a mouth rinse. In accordance with some embodiments, the gel or foam is formulated as an aerosol spray or hydrogel. In accordance with some embodiments, the composition described by the present disclosure is presented on or in a solid support. In accordance with some embodiments, the solid support contains cotton fibers. In accordance with some embodiments, the solid support is a bandage or fabric mask.

[020] In accordance with some embodiments, the present invention relates to a solid support containing a composition described herein. In accordance with some embodiments, the kit further includes one or more elastic bands. In accordance with some embodiments, the composition is formulated as a foam, gel, or liquid.

[021] In accordance with some embodiments, the solid support contains cotton fibers. In accordance with some embodiments, the solid support is a bandage (eg, a non-adhesive bandage) or a mask (eg, a cloth face mask).

[022] In some aspects, the present invention provides a kit comprising a composition described herein and a non-adhesive wound dressing. In accordance with some embodiments, the non-adhesive wound dressing contains cotton fibers.

[023] In some aspects, the present invention provides a kit comprising a composition described herein and one or more antiviral agents effective for treating herpes simplex virus.

[024] In accordance with some embodiments, one or more antiviral agents is selected from acyclovir, valacyclovir, and famciclovir. In accordance with some embodiments, the herpes simplex virus is herpes simplex labialis (HSL). In accordance with some embodiments, the composition, antiviral agent, or combination thereof is formulated for topical use.

[025] In some aspects, the present invention provides a method of treating oral lesions in a subject (e.g., a human subject), the method comprising administering a composition described herein to a subject having one or more oral lesions .

[026] In accordance with some embodiments, the subject has or is suspected of having mucositosis. In accordance with some embodiments, the subject has previously been subjected to chemotherapy, radiation therapy, or a combination of chemotherapy and radiation therapy. In accordance with some embodiments, the subject has undergone maxillofacial surgery.

[027] In accordance with some embodiments, the composition is applied directly to the oral cavity of a subject. In some embodiments, application is carried out using a mouth spray or mouth rinse. In accordance with some embodiments, the composition is administered to a subject more than once per day (eg, 2, 3, 4, or more times per day). In accordance with some embodiments, the composition is administered to a subject before a meal, after a meal, or both before and after a meal. In accordance with some embodiments, use of the composition inhibits the formation and/or growth of a bacterial biofilm.

[028] In some aspects, the present invention provides a method of applying a composition described herein to the skin of a subject in need thereof.

[029] In accordance with some embodiments, the subject's skin has been subjected to a cosmetic procedure. In accordance with some embodiments, the cosmetic procedure is a laser skin peel.

[030] In accordance with some embodiments, the subject's skin is damaged. In accordance with some embodiments, the damaged skin is a wound (eg, surgical incision, etc.), ulcer, or blister.

[031] In accordance with some embodiments, the use involves bringing the subject's skin into contact with a solid support containing the composition. In accordance with some embodiments, the solid support is a non-adhesive bandage or fabric face mask.

[032] In accordance with some embodiments, the composition is applied topically. In accordance with some embodiments, application occurs at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times per day. In accordance with some embodiments, application occurs at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times per week.

[033] In accordance with some embodiments, the subject has or is suspected of having diabetes mellitus (eg, type I diabetes mellitus or type II diabetes mellitus). In some embodiments, the subject has a chronic wound resulting from diabetes mellitus or damaged skin resulting from insufficient blood flow, such as varicose veins or a bedsore. In accordance with some embodiments, the subject has or is suspected of having a diabetic ulcer, such as a diabetic foot ulcer.

[034] In some embodiments, the subject is infected or suspected of being infected with herpes simplex virus. In accordance with some embodiments, the virus is herpes simplex labialis.

[035] In some aspects, the present invention provides a method for regenerating a peripheral nerve terminal in a subject in need, the method comprising administering to the subject a first composition comprising a biologically active extract of Melissa officinalis and a second composition containing methylcobalamin and/or cyanocobalamin.

[036] In accordance with some embodiments, the first composition and the second composition are administered as a single composition. In accordance with some embodiments, the first and/or second composition is applied topically.

[037] In accordance with some embodiments, the subject's skin is damaged. In accordance with some embodiments, the damaged skin is a wound, ulcer, or blister. In accordance with some embodiments, the subject is infected or suspected of being infected with herpes simplex virus. In accordance with some embodiments, the herpes simplex virus is herpes simplex labialis.

Brief description of drawings

[038]

In fig. Figure 1 shows representative data on the effectiveness of tissue repair formulations against 3-day MRSA S. aureus biofilms. Both daily and single applications were evaluated. Please note that data are presented in terms of surviving microorganisms. Ex = filler.

In fig. Figure 2 shows representative data on the effectiveness of tissue repair formulations against 3-day P. aeruginosa biofilms. Both daily and single applications were evaluated. Please note that data are presented in terms of surviving microorganisms. Ex = filler.

In fig. Figure 3 shows representative data on the effectiveness of tissue repair formulations against 3-day C. albicans biofilms. Both daily and single applications were evaluated. Please note that data are presented in terms of surviving microorganisms. Ex = filler.

In fig. Figure 4 shows schematic representations of wound sites on the back of a pig randomly selected for each treatment.

In fig. 5A-5H are photographs showing the healing process of penetrating surgical wounds on pig skin with various treatments and controls. In fig. 5A shows data for composition A. FIG. 5B shows data for composition B. FIG. 5C shows data for composition C. FIG. 5D shows data for composition D. FIG. 5E shows data for the saline control (SAL). In fig. 5F shows data for Plurogel gel. In fig. 5G shows data for Medihoney gel. In fig. Figure 5H shows data for Amerigel gel.

In fig. 6 shows the percentage reduction in wound area when treated with wound healing compositions and saline control. A - composition A; B - composition B; C - composition C; D - composition D; PG - Plurogel; MN - Medihoney; AMG - Amerigel; SAL - saline solution.

Detailed Disclosure of the Present Invention

[039] Chronic wounds that do not progress through the normal wound healing pattern but remain in a state of chronic inflammation typically present a protease-rich and pro-oxidatively hostile microenvironment. This microenvironment typically promotes the destruction of growth factors and excess production of reactive oxygen species (ROS), leading to greater tissue damage and delayed tissue repair. Microorganisms present in the wound also contribute to impaired tissue repair.

[040] The present invention relates to drug delivery systems and drug compositions for promoting wound healing and tissue formation. The present invention is based in part on drug delivery compositions (systems) containing one or more of the following: a surfactant (eg, a surfactant), a hydrating agent (eg, a hygroscopic agent), and a carbonate buffer that maintains the pH of the composition in the range of approximately pH 8.0 to pH 9.5. In accordance with some embodiments, the composition further comprises one or more biologically active agents.

[041] Without wishing to be bound by any particular theory, when applied to damaged tissue (e.g., skin, mucous membrane, etc.) of a subject, the compositions described herein create a microenvironment that promotes tissue healing and regeneration while providing 1) molecular sanitation of the wound; 2) proper hydration; and 3) anti-inflammatory and antimicrobial effects. For example, in accordance with some embodiments, the compositions described herein assist in the removal of bacterial biofilm by disrupting the structure of the exopolymer polysaccharide framework. In accordance with some embodiments, the properties and/or activities of the compositions described herein are unexpected from the perspective of currently available wound healing compositions, which typically have pH values ranging from neutral (e.g., approximately pH 7) to acidic (e.g. , approximately pH 5.0-6.0).

[042] Accordingly, in some aspects, the present invention provides a composition comprising a drug delivery system comprising a carbonate buffer solution, at least two different biological surfactants, at least one hygroscopic agent, and at least one antioxidant ; and at least one biologically active agent, the composition having a pH in the range from about 8.0 to about 9.5.

Means for adjusting surface tension

[043] The present invention relates in part to compositions containing one or more surface tension adjusting agents. In accordance with some embodiments, the surface tension adjusting agent is a surfactant.

[044] Typically, the composition described herein may contain one or more surfactants (for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more than 10 surfactants) . In accordance with some embodiments, each surfactant of the composition is selected from a biological surfactant, an anionic surfactant, a zwitterionic surfactant, and a nonionic surfactant. In accordance with some embodiments, the composition described herein does not have (eg, does not contain) a surfactant.

[045] In accordance with some embodiments, the at least one surfactant is a biological surfactant. As used herein, the term “biological surfactant” refers to surfactant biomolecules produced by microorganisms (eg, certain bacterial cells) that reduce the surface tension of cells (eg, the surface tension of cell membranes, such as mammalian cell membranes).

[046] Examples of biological surfactants include, but are not limited to, glycolipids (e.g., rhamnolipids, sophorolipids, trehalolipids, cellobiolipids, mannosylerythritol lipids, etc.), lipopeptides (e.g., surfactin, plipastatin, bacillomycin, fengycin, subtilisin, gramicidin, polymyxins, etc.) and polymeric biological surfactants (for example, emulsan, biodispersan, liposan, mannan-lipid-protein, carbohydrate-lipid-protein, etc.). In accordance with some embodiments, the biological surfactant is selected from a glycolipid, a lipopeptide, and a polymeric biological surfactant.

[047] In accordance with some embodiments, the biological surfactants disclosed herein are one or more biological surfactants selected from glycolipids. Glycolipids are carbohydrates linked by an ester group to long-chain aliphatic acids or hydroxyaliphatic acids. Examples of glycolipids include, but are not limited to, rhamnolipids, trehalolipids, and sophorolipids. In accordance with some embodiments, the glycolipids are produced by bacterial cells of Pseudomonas aeruginosa.

[048] As used herein, the term “rhamnolipid” refers to glycolipids in which one or two rhamnose molecules are associated with one or two hydroxydecanoic acid molecules. In some embodiments, the rhamnolipid is 3-[3-[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxydecanoyloxy]decanoic acid, also referred to herein as "rhamnolipid RI".

[049] As used herein, the term “sophorolipid” refers to glycolipids that are produced by yeast and consist of a dimeric carbohydrate sophorolipid linked by a glycosidic bond to a long-chain hydroxyl fatty acid. Sophorolipids can be a mixture of at least six to nine different hydrophobic sophorolipids. In accordance with some embodiments, the sophorolipid is the lactone form of the sophorolipid. According to some embodiments, the sophorolipid is (E)-17-[(2R,3R,4S,5S,6R)-6-(acetyloxymethyl)-3-[(2S,3R,4S,5S,6R)- 6-(acetyloxymethyl)-3,4,5-trihydroxyoxan-2-yl]oxy-4,5-dihydroxyoxan-2-yl]oxyoctadec-9-enoic acid.

[050] In accordance with some embodiments, the composition contains a combination of surfactants, such as rhamnolipid and sophorolipid (eg, at least one rhamnolipid and at least one sophorolipid). The relative amounts (eg, ratio) of surfactants (eg, rhamnolipid and sophorolipid) in the composition may vary. In some embodiments, the ratio of rhamnolipid to sophorolipid ranges from about 1:9 to about 9:1 (e.g., any ratio from 1:9 to 9:1, e.g., 1:1, 2:8, 8: 2, 7:3, 3:7, 6:4, 4:6, etc.).

[051] The ratio or amount of biological surfactants in the composition (for example, the ratio of rhamnolipid to sophorolipid) can be measured, for example, in % by weight (w/w), % by volume (v/v), molar concentration etc. relative to the entire composition.

[052] The total amount of biological surfactant in the composition may vary. In accordance with some embodiments, the total amount of biological surfactant in the composition ranges from about 0.1% (w/w) to about 8% (w/w). According to some embodiments, the total amount of biological surfactant in the composition is about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0. 7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1, 7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2, 7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3, 7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4, 7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5, 7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6, 7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7, 7%, 7.8%, 7.9% or approximately 8.0%. In accordance with some embodiments, the composition described herein does not contain a biological surfactant.

[053] In solution, depending on concentration and temperature, surfactants structurally form a spherical micelle called a "unimer". The structure of the micelles changes over time, contracting and expanding, forming a multimer. In accordance with some embodiments, the unimers are capable of permanently trapping foreign substances in the wound, creating a rinsing effect. Because the surfactant reduces the surface tension between the wound bed and the cleansing fluid, the cleansing fluid comes into close contact with the wound bed. This facilitates the separation of loose, nonviable tissue and microbial particles from the viable wound bed, which inhibits (eg, prevents) biofilm formation and promotes the destruction of older, more recalcitrant biofilms. In some embodiments, surfactants disrupt and prevent re-formation of biofilm following wound debridement. In accordance with some embodiments, the presence of highly stretchable biomolecules, such as rhamnolipids and sophorolipid biological surfactants, in the compositions described herein promotes biochemical microdebridization, which results in effective destruction of the polysaccharide matrix of the bacterial biofilm and removal of devitalized and necrotic tissue that appears due to cellular damage.

[054] In accordance with some embodiments, the compositions described herein contain one or more anionic surfactants. Examples of anionic surfactants include soaps, alkylbenzene sulfonates, alkyl sulfonates, alkyl sulfonates, alkyl sulfates, fluorinated fatty acid salts, silicones, fatty alcohol sulfates, polyoxyethylene fatty alcohol ether sulfates, α-olefin sulfonate, polyoxyethylene fatty alcohol phosphate ether, alkyl alcohol amide , alkyl sulfonic acid acetamide, alkyl succinate sulfonate salts, alkyl benzene sulfonates of amino alcohols, naphthenates, alkyl phenol sulfonate and polyoxyethylene monolaurate.

[055] The total amount of anionic surfactant in the composition may vary. In accordance with some embodiments, the total amount of anionic surfactant in the composition ranges from about 0.1% (w/w) to about 8% (w/w). According to some embodiments, the total amount of biological surfactant in the composition is about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0. 7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1, 7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2, 7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3, 7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4, 7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5, 7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6, 7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7, 7%, 7.8%, 7.9% or approximately 8.0%. In accordance with some embodiments, the composition described herein does not have (eg, does not contain) an anionic surfactant.

[056] In accordance with some embodiments, the compositions described herein contain one or more nonionic surfactants. Examples of the nonionic surfactant include polyethylene glycol alkyl ethers (Brij), octaethylene glycol monododecyl ether, pentaethylene glycol monododecyl ether, polypropylene glycol alkyl ethers, glucoside alkyl ethers, decyl glucoside, lauryl glucoside, octyl glucoside, octylphene polyethylene glycol yl ethers, polyethylene glycol alkylphenyl ethers , glycerol alkyl esters, glyceryl laurate, polyoxyethylene glycol sorbitan alkyl esters, sorbitan alkyl esters, cocamide MEA, cocamide DEA, block copolymers of polyethylene glycol and polypropylene glycol: poloxamers, polyethoxylated fatty amine (POEA).

[057] The total amount of nonionic surfactants in the composition may vary. In accordance with some embodiments, the total amount of nonionic surfactants in the composition ranges from about 0.1% (w/w) to about 8% (w/w). According to some embodiments, the total amount of nonionic surfactants in the composition is about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0. 7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1, 7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2, 7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3, 7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4, 7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5, 7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6, 7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7, 7%, 7.8%, 7.9% or approximately 8.0%. In accordance with some embodiments, the composition described herein does not have (eg, does not contain) a nonionic surfactant.

[058] The composition may also contain one or more zwitterionic surfactants. Examples of zwitterionic surfactants include, but are not limited to, synthetic zwitterionic surfactants (eg, one or more hydroxysultaines) and/or naturally occurring zwitterionic surfactants (eg, one or more betaines, phosphatidylcholines, and/or lecithin components). In accordance with some embodiments, the zwitterionic surfactant is cocamidopropyl betaine.

[059] The total amount of zwitterionic surfactants in the composition may vary. In accordance with some embodiments, the total amount of zwitterionic surfactants in the composition ranges from about 0.1% (w/w) to about 8% (w/w). According to some embodiments, the total amount of zwitterionic surfactants in the composition is about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0. 7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1, 7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2, 7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3, 7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4, 7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5, 7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6, 7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7, 7%, 7.8%, 7.9% or approximately 8.0%. In accordance with some embodiments, the composition described herein does not have (eg, does not contain) a zwitterionic surfactant.

pH/buffer systems

[060] Current skin care compositions typically have a pH that is close to the natural pH of human skin (e.g., pH 6.8-7.0) or slightly more acidic than the pH of human skin (e.g., pH 5. 0-7.0). The present disclosure is based in part on compositions having a pH greater than 7 (eg, more basic or alkaline than neutral), which create a microenvironment that promotes tissue healing and regeneration. In accordance with some embodiments, the healing and/or regenerative activity of the compositions described herein is unexpected in light of the more basic pH of the compositions compared to typical wound care products.

[061] The pH value of the composition described herein is in the range from about 7.5 to about 10.0. In accordance with some embodiments, the pH value of the composition described herein is in the range from about 8.0 to about 9.5. In accordance with some embodiments, the pH of the composition described herein is approximately 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9, 6, 9.7, 9.8, 9.9 or 10.0).

[062] Methods for measuring the pH of a composition are known, for example, using a pH meter, electrode measurement (eg, glass electrodes, reference electrodes, combination electrodes, etc.), colorimetric measurement, etc. Typically, pH is measured at room temperature (eg, 18°C to 24°C). However, it should be understood that pH can be measured at other temperatures (for example, below 10°C, above 25°C, etc.).

[063] The pH of the composition can be maintained using a buffer system. Examples of buffer systems include, but are not limited to, carbonate buffer systems, phosphate buffer systems, protein buffer systems, etc. The composition may include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more buffer systems.

[064] The present disclosure is based in part on compositions containing a carbonate buffer system. Generally, the term "carbonate buffer system" refers to a solution containing a weak acid (such as carbonic acid) and its conjugate base (such as a bicarbonate anion) that buffers changes in the pH of the solution. The conjugate base (e.g. bicarbonate anion) can be derived from any conjugate salt of carbonic acid, for example: sodium bicarbonate (NaHCO ₃ ), potassium bicarbonate (KHCO ₃ ), cesium bicarbonate (CsHCO ₃ ), magnesium bicarbonate (Mg(HCO ₃ ) ₂ ), calcium bicarbonate (Ca(HCO ₃ ) ₂ ) and ammonium bicarbonate (NH ₅ CO ₃ ). In accordance with some embodiments, the carbonate buffer system includes sodium bicarbonate (NaHCO ₃ ). In accordance with some embodiments, the carbonate buffer system includes ammonium bicarbonate (NH ₅ CO ₃ ).

[065] In addition to pH buffering, certain buffer systems (eg, Na-based and NH ⁴ -based buffer systems) increase the concentration of Na ⁺ or NH ₄ ⁺ ions in the composition or in the microenvironment created when the composition comes into contact with damaged tissue (for example, a wound , blister, ulcer, etc.). Without wishing to be bound by any particular theory, large ions (such as sodium ions) bind to certain molecules on the surface of mammalian cells, such as heparin sulfate (HS), and prevent pathogens from entering cells, for example as described by Rabenstein et al . (2002) Nat. Prod. Rep. 19:312-331. In accordance with some embodiments, the binding of large ions to the cell membrane also affects the signaling of growth factors such as FGF and EGF, which are important for skin regeneration.

[066] In accordance with some embodiments, the compositions described herein contain (or provide) a quantity of ions (e.g., Na ⁺ or NH ₄ ⁺ ions) in the wound microenvironment sufficient to disrupt the association of microorganisms, such as bacteria and viruses, with proteoglycans (eg HS, etc.) on the surface of target cells.

[067] In accordance with some embodiments, the composition described herein contains from about 0.5% (w/w) to about 10% (w/w) bicarbonate salt (eg, sodium bicarbonate). According to some embodiments, the composition contains from about 2% to about 5% (w/w) bicarbonate salt (e.g., sodium bicarbonate), such as about 2%, 2.1%, 2.2%, 2 ,3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3 ,3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0, 4.1%, 4.2%, 4, 3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9% or 5% (w/w) sodium bicarbonate. In accordance with some embodiments, the composition described herein does not have (eg, does not contain) a carbonate buffer system.

Hydrating agents

[068] The compositions of the present invention may contain one or more (eg, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) hydrating agents. "Hydrating agent" or "hygroscopic agent" generally refers to a molecule or molecules that, through either absorption or adsorption, attract and/or retain water molecules from the environment. Examples of hygroscopic agents include polymers such as cellulose polymers, glycol polymers, glycosaminoglycans, mucopolysaccharides, etc.

[069] In accordance with some embodiments, the hygroscopic agent is a non-solubilizing agent (eg, a molecule that absorbs enough water from its surroundings to form an aqueous solution). Examples of dissolving agents include salts (eg, calcium chloride, magnesium chloride, zinc chloride, ferric chloride, carnallite, potassium carbonate, potassium phosphate, ferric citrate, ammonium nitrate, potassium hydroxide and sodium hydroxide).

[070] In accordance with some embodiments, the composition contains one or more cellulose polymers, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more cellulose polymers. In accordance with some embodiments, the composition does not have (eg, does not contain) a hydrating agent. Typically, cellulose polymers contain two or more repeating subunits (eg, polymer subunits) of glucose. Examples of cellulose polymers include methylcellulose, hydroxymethylcellulose, hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC), hydroxyethylcellulose and carboxymethylcellulose (CMC). In accordance with some embodiments, at least one of the cellulose polymers is hydroxyethylcellulose or carboxymethylcellulose.

[071] The amount of one or more cellulose polymers in the composition can vary. In accordance with some embodiments, the composition contains from about 0.1% (w/w) to about 5% (w/w) cellulose polymer. According to some embodiments, the composition contains approximately 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9% or 5.0% (w/w) cellulose polymer.

[072] In accordance with some embodiments, the composition contains one or more glycol polymers, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more glycol polymers. Typically, glycol polymers contain two or more repeating subunits (eg, polymer subunits) of a polyether, such as ethylene oxide. In accordance with some embodiments, the glycol polymer is polyethylene glycol (PEG, also referred to as PEG and POE). Additional examples of glycol polymers include methoxy poly(ethylene glycol) and polypropylene glycol (PPG). In accordance with some embodiments, at least one of the glycol polymers is PEG.

[073] The number of polymer subunits in a glycol polymer (eg, PEG) can vary. In accordance with some embodiments, the glycol polymer (eg, PEG) contains from 2 to 10,000,000 polymer subunits (eg, any integer from 2 to 10,000,000, inclusive). In accordance with some embodiments, the glycol polymer (eg, PEG) is composed of more than 10,000,000 polymer subunits.

[074] In accordance with some embodiments, a glycol polymer, such as a PEG polymer, is described by its molecular weight (eg, measured in g/mol). In some embodiments, the glycol polymer is PEG 400 (e.g., a 400 dalton average molecular weight PEG polymer), PEG 500 (e.g., a 500 dalton average molecular weight PEG polymer), PEG 1000 (e.g., an average molecular weight PEG polymer). molecular weight 1000 dalton), PEG 3500 (for example, PEG polymer with an average molecular weight of 3500 dalton), PEG 4000 (for example, PEG polymer with an average molecular weight 4000 dalton), PEG 10000 (for example, PEG polymer with an average molecular weight 10000 dalton) , PEG 50000 (for example, a PEG polymer with an average molecular weight of 50,000 daltons), PEG 100,000 (for example, a PEG polymer with an average molecular weight of 100,000 daltons), or PEG 1,000,000 (for example, a PEG polymer with an average molecular weight of 1,000,000 daltons).

[075] The geometry (eg, structure) of a glycol polymer such as PEG can vary. In accordance with some embodiments, the glycol polymer is a linear polymer (eg, linear PEG). In accordance with some embodiments, the glycol polymer is a branched polymer (eg, a branched PEG, such as a "star PEG"). In accordance with some embodiments, the glycol polymer is a comb PEG (eg, multiple PEG chains grafted onto a polymer backbone).

[076] The amount of one or more glycol polymers (eg, PEG) in the composition can vary. In accordance with some embodiments, the composition contains from about 0.1% (wt./wt.) to about 15% (wt./wt.) glycol polymer. In accordance with some embodiments, the composition contains approximately 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9% or 5.0% (w/w) glycol polymer. In accordance with some embodiments, the composition contains approximately 10%, 11%, 12%, 13%, 14%, or 15% (w/w) glycol polymer. According to some embodiments, the composition contains no more than 15%-20% (w/w) (e.g., no more than 15%, 16%, 17%, 18%, 19%, or 20% (w/w) .)) glycol polymer.

[077] In accordance with some embodiments, the composition contains one or more glycosaminoglycans, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more glycosaminoglycans. In general, glycosaminoglycans are long, unbranched polysaccharides containing two or more repeating disaccharide subunits (eg, polymer subunits), such as an amino sugar and a uronic sugar (eg, glucuronic acid, iduronic acid) or galactose. Examples of glycosaminoglycans include heparin (Hep)/heparin sulfate (HS), chondroitin sulfate (CS)/dermatan sulfate (DS) and hyaluronic acid (HA), etc. In accordance with some embodiments, at least one of the glycosaminoglycans is hyaluronic acid (HA).

[078] The number of polymer subunits and therefore the molecular weight (eg, weight average molecular weight) of hyaluronic acid (HA) can vary. In accordance with some embodiments, hyaluronic acid (HA) has a molecular weight of about 5,000 to 20,000,000 daltons. In accordance with some embodiments, the number of disaccharide polymer subunits in the HA ranges from about 2 polymer subunits to about 50,000 polymer subunits (eg, any integer from 2 to 50,000, inclusive).

[079] The amount of one or more glycosaminoglycans in the composition may vary. In accordance with some embodiments, the composition contains from about 0.1% (wt./wt.) to about 5% (wt./wt.) glycosaminoglycans. According to some embodiments, the composition contains approximately 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9% or 5.0% (w/w) glycosaminoglycans.

[080] In accordance with some embodiments, the drug delivery composition described herein comprises water. The amount of water in the composition may vary. According to some embodiments, the composition contains at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% (w/w) water.

Biologically active agents

[081] The compositions described herein may contain one or more biologically active agents. Examples of biologically active agents include antimicrobial agents (e.g., antibacterial agents, antiviral agents, antiparasitic agents, etc.), cytotoxic agents, anticancer agents, free radical scavengers, antioxidants, receptor ligands (e.g., molecules that induce or inhibit signaling in a cage, etc.) etc. The biologically active agent may be a small molecule (eg, chemical), peptide, protein, polypeptide, nucleic acid (eg, DNA, RNA, etc.) or biologically active extract. In accordance with some embodiments, the composition contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 biologically active funds. In accordance with some embodiments, the composition contains more than 20 biologically active agents.

[082] In accordance with some embodiments, the biologically active agent is an antimicrobial agent, such as an antibacterial agent, an antiviral agent, or an antiparasitic agent. Typically, the antimicrobial agent may be a small molecule (eg, chemical), peptide, protein, polypeptide, nucleic acid (eg, DNA, RNA, etc.). Examples of antimicrobial agents include, but are not limited to, small molecules derived from bacteria and fungi (e.g., amoxicillin, doxycycline, cephalexin, ciprofloxacin, metronidazole, etc.), small molecules derived from plants (e.g., tannins, flavones, phenols, alkaloids) etc.), and antimicrobial peptides (for example, maxamine, dermicidin, mecropin, andropin, etc.). In some embodiments, the bioactive agent is a plant-derived antimicrobial agent, such as baicalin (eg, extracted from Scutellaria baicalensis or Scutellaria lateriflora) or andrographolide (eg, extracted from Andrographispaniculata). In accordance with some embodiments, the composition described herein does not have a chemical or elemental antimicrobial agent, such as silver (e.g., silver-based compounds such as silver sulfadiazine) or zinc (e.g., zinc-based compounds such as zinc oxide ).

[083] In accordance with some embodiments, the biologically active agent is a cytotoxic agent. Examples of cytotoxic agents include, but are not limited to, alkylating agents (eg, cyclophosphamide, nitrosoureas, etc.), anthracyclines (eg, doxorubicin, daunorubicin, etc.), taxanes (eg, taxol, paclitaxel, etc.), inhibitors HD AS, nucleotide analogues (eg, gemcitabine, etc.), platinum-based compounds (eg, cisplatin, etc.) and vinca alkaloids (eg, vinblastine, etc.).

[084] In accordance with some embodiments, the biologically active agent is an antioxidant or free radical scavenger. Examples of antioxidants and free radical scavengers include, but are not limited to, certain enzymes (e.g., superoxide dismutase, glutathione peroxidase, glucose oxidase catalase, etc.), carotenoids (e.g., astaxanthin, beta-carotene, tocopherol, etc.), phenolic compounds (e.g. , plant-derived polyphenols such as anthocyanins, flavan-3-ols (catechin), flavonols (e.g. quercetin and rutin), cinnamic acids (e.g. S-glutathionyl caftaric acid), caffeic acid phenethyl ester (CAPE), chalcones, isoflavonoids ( for example, 7-O-methylvestitol, medicarpine and 3,4,2',3'-tetrahydrochalcone), etc.

[085] In accordance with some embodiments, the biologically active agent is a cell signaling molecule, such as a receptor ligand. Examples of cell signaling molecules include, but are not limited to, neurotransmitters (e.g., GABA, glutamate, acetylcholine, serotonin, dopamine, etc.), cytokines (e.g., IL-4, IL-15, TNFα, IFNγ, etc.), hormones (eg, estrogen, etc.), small molecules (eg, nitric oxide, etc.), certain peptides (eg, neuropeptides, growth factors, etc.), etc.

[086] As used herein, the term “bioactive extract” refers to a composition containing one or more biologically active agents that have been extracted (e.g., isolated) from an organic source, such as one or more plants or plant products, animals or products of animal origin, insects or products derived from insects, microorganisms or products derived from microorganisms, etc. In general, biologically active extracts can be obtained using any suitable method, for example, solvent extraction methods (eg, alcohol extraction, hydrocarbon extraction, etc.), maceration extraction methods, ultrasonic extraction (eg, sonication), extraction with using microwaves (MAE), etc. However, one skilled in the art will appreciate that the appropriate extraction method will depend on the type of material from which the bioactive agents are to be isolated and will select an appropriate extraction method.

[087] The amount of biologically active agent or extract in the composition may vary. In accordance with some embodiments, the concentration of the biologically active agent or biologically active extract in the composition is in the range of about 0% wt./wt. (none) to about 20% w/w (e.g., about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12% , 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20%) of the total weight of the composition.

Biologically active propolis extracts

[088] In accordance with some embodiments, the composition contains a biologically active extract derived from propolis. The term "propolis" generally refers to a resinous material produced by honey bees (e.g. Apis and other species) that contains a variety of chemicals and molecules, including varieties of wax, fatty acids, pollen, aromatic oils, and certain bioactive molecules (e.g. , flavonoids, isoflavinoids, chalcones, pterocarpanes, etc.). Propolis can be found in several regions of the world, including North and Central America (e.g. USA, Canada, Mexico, Cuba, etc.), South America (e.g. Brazil, Colombia, Chile, etc.), Asia (eg China) and New Zealand. In accordance with some embodiments, the biologically active propolis extract is obtained from a blend (eg, mixture) containing at least one Brazilian propolis variety. Biologically active propolis extract can be obtained using any suitable extraction method, for example, ethanol/glycol extraction or supercritical CO ₂ extraction.

[089] Categorization of propolis sources may vary. For example, Brazilian propolis was initially classified by geographic origin and/or physicochemical properties, and at least 12 sources of propolis were identified, including but not limited to five in the southern Brazilian group (group 3), one in the south-Brazil group eastern Brazil (group 12) and six in the northeastern Brazil group (group 6). However, when classified by botanical origin, three sources of Brazilian propolis were identified for the same propolis varieties: poplar trees (e.g. Populus sp.), baccharis (e.g. Baccharis sp., such as Baccharis dracunculifolia, etc.) and shrubs (eg Hyptis sp. such as Hyptis divaricata'). Additional sources of propolis are also known, for example as described by Park et al. (2002) J. Agric. Food Chem 50: 2502-2506, and Dezmirean et al. (2017) J. Apicultural Res 56(5): 588-597. In some embodiments, propolis is categorized by its color, for example: red propolis, green propolis, brown propolis, etc.

[090] "Red propolis" refers to propolis obtained botanically from plants such as Dalbergia ecastophyllum(D. ecastophyllum) (L) Taub. (Fabaceae), which are commonly known in Brazil as "rabo-de-bugio". In accordance with some embodiments, red propolis is obtained from a region selected from Brazil, Cuba, Mexico, China and Nigeria. Sources of red propolis are described, for example, by Corbellini Rufatto et al. (2017) Asian Pacific Journal of Tropical Biomedicine 7(7): 591-598.

[091] “Green propolis” refers to propolis obtained botanically from plants such as Baccharis dracunculifolia DC (Asteraceae), for example, as described in Lopes Machado et al, (2012) Evid Based Complement Alternat Med. 2012:157652. In some embodiments, green propolis is obtained from Brazil (eg, southeastern Brazil, such as the state of Bahia, the state of Minas Gerais, the state of Sao Paulo, or the state of Parana).

[092] "Brown propolis" refers to propolis obtained botanically from plants such as Populus spp. (P. alba, P. nigra, P. tremuld) or Clusia spp. In accordance with some embodiments, brown propolis is obtained from a region selected from Brazil, Venezuela, Cuba and Europe.

[093] The ratio of each type of propolis in the mixture from which the biologically active propolis extract is obtained may vary. For example, in some embodiments, the propolis mixture contains green and brown propolis in a ratio of about 9:1 to about 1:9 (e.g., 1:1, 1:2, 1:3, 1:4, 1:5 , 1:6, 1:7, 1:8, 1:9, 2:1, 2:3, 2:4, 2:5, 2:6, 2:7, 2:8, 2:9, 3 :1, 3:2, 3:4, 3:5, 3:6, 3:7, 3:8, 3:9, 4:1, 4:2, 4:3, 4:5, 4:6 , 4:7, 4:8, 4:9, 5:1, 5:2, 5:3, 5:4, 5:6, 5:7, 5:8, 5:9, 6:1, 6 :2, 6:3, 6:4, 6:5, 6:7, 6:8, 6:9, 7:1, 7:2, 7:3, 7:4, 7:5, 7:6 , 7:8, 7:9, 8:1, 8:2, 8:3, 8:4, 8:5, 8:6, 8:7, 8:9, 9:8, 9:7, 9 :6, 9:5, 9:4, 9:3, 9:2, 9:1, etc.). In some embodiments, the propolis mixture contains green and red propolis in a ratio of about 9:1 to about 1:9 (e.g., 1:1, 1:2, 1:3, 1:4, 1:5, 1 :6, 1:7, 1:8, 1:9, 2:1, 2:3, 2:4, 2:5, 2:6, 2:7, 2:8, 2:9, 3:1 , 3:2, 3:4, 3:5, 3:6, 3:7, 3:8, 3:9, 4:1, 4:2, 4:3, 4:5, 4:6, 4 :7, 4:8, 4:9, 5:1, 5:2, 5:3, 5:4, 5:6, 5:7, 5:8, 5:9, 6:1, 6:2 , 6:3, 6:4, 6:5, 6:7, 6:8, 6:9, 7:1, 7:2, 7:3, 7:4, 7:5, 7:6, 7 :8, 7:9, 8:1, 8:2, 8:3, 8:4, 8:5, 8:6, 8:7, 8:9, 9:8, 9:7, 9:6 , 9:5, 9:4, 9:3, 9:2, 9:1, etc.). In some embodiments, the propolis mixture contains brown and red propolis in a ratio of about 9:1 to about 1:9 (e.g., 1:1, 1:2, 1:3, 1:4, 1:5, 1 :6, 1:7, 1:8, 1:9, 2:1, 2:3, 2:4, 2:5, 2:6, 2:7, 2:8, 2:9, 3:1 , 3:2, 3:4, 3:5, 3:6, 3:7, 3:8, 3:9, 4:1, 4:2, 4:3, 4:5, 4:6, 4 :7, 4:8, 4:9, 5:1, 5:2, 5:3, 5:4, 5:6, 5:7, 5:8, 5:9, 6:1, 6:2 , 6:3, 6:4, 6:5, 6:7, 6:8, 6:9, 7:1, 7:2, 7:3, 7:4,7:5, 7:6, 7 :8, 7:9, 8:1, 8:2, 8:3, 8:4, 8:5, 8:6, 8:7, 8:9, 9:8, 9:7, 9:6 , 9:5, 9:4, 9:3, 9:2, 9:1, etc.). In some embodiments, the propolis mixture contains green, brown, and red propolis in a ratio of from about 9:1:1 to about 1:9:1, to about 1:1:9 (e.g., 1:1:1, 1 :2:1, 2:1:1, 1:1:2, etc.).

[094] The amount of biologically active propolis extract in the composition of the present invention may vary. In accordance with some embodiments, the composition described herein (eg, a drug delivery composition) contains from about 0.1% wt./wt. up to approximately 20% w/w. biologically active propolis extract. In accordance with some embodiments, the composition contains from about 0.1% wt./wt. to approximately 4% w/w. biologically active propolis extract. In accordance with some embodiments, the composition contains from about 1% to about 5% wt./wt. biologically active propolis extract. In accordance with some embodiments, the composition contains from about 3% to about 7% wt./wt. biologically active propolis extract. In accordance with some embodiments, the composition contains from about 5% to about 20% wt./wt. biologically active propolis extract.

[095] Extracts derived from propolis (eg, one or more types of propolis, such as a blend of brown propolis, green propolis, red propolis, or any combination thereof) may contain a number of biologically active agents. In accordance with some embodiments, the biologically active extract obtained from propolis contains at least one of the following: fatty and phenolic acids and esters, substituted phenolic esters, flavonoids (e.g., flavones, flavanones, flavonols, dihydroflavonols, chalcones, and etc.), mono-, sesqui-, di- and triterpenes, steroids, aromatic aldehydes and alcohols, naphthalene and stilbene derivatives, caffeoylquinic acid derivatives, lignans, coumarins, prenylated and coumarin derivatives. Biologically active molecules derived from propolis are known in the art, for example as described by Schindler Machado et al. (2016) Evid Based Complement Alternat Med. 2016:6057650, and Trusheva et al. (2006) Evid Based Complement Alternat Med. 3 (2): 249-254, and Huang et al. (2014) Molecules 19:19610–19632.

[096] The present disclosure is based in part on biologically active propolis extracts containing one or more (eg, 1, 2, 3, 4, 5, 6, etc.) antioxidants. In accordance with some embodiments, the one or more antioxidants are flavonoids (eg, isoflavonoids, etc.). In some embodiments, the flavonoid bioactives in the bioactive propolis extract promote the removal of reactive oxygen species (ROS) generated by DNA damage in irradiated cellular elements, thereby causing a reduction in the destruction of the extracellular matrix of the oral mucosa. In some embodiments, flavonoids are present in the bioactive propolis extract at a concentration of from about 100 μg/ml to about 500 μg/ml (e.g., about 100 μg/ml, about 150 μg/ml, about 200 μg/ml, about 250 µg/ml, approximately 300 µg/ml, approximately 350 µg/ml, approximately 400 µg/ml, approximately 450 µg/ml, etc.), as measured by gravimetric methods. In some embodiments, flavonoids are present in the bioactive propolis extract at a concentration of from about 100 μg/ml to about 200 μg/ml, as measured by gravimetric methods. In some embodiments, flavonoids are present in the bioactive propolis extract at a concentration of from about 150 μg/ml to about 300 μg/ml, as measured by gravimetric methods. In some embodiments, flavonoids are present in the bioactive propolis extract at a concentration of from about 250 μg/ml to about 400 μg/ml, as measured by gravimetric methods.

[097] In accordance with some embodiments, the biologically active propolis extract contains one or more lipids, one or more waxes, or a combination of one or more lipids and one or more waxes. In some embodiments, the total concentration of lipids and wax species in the bioactive propolis extract ranges from about 25 μg/ml to about 750 μg/ml (e.g., about 25 μg/ml, 50 μg/ml, 75 μg/ml). ml, 100 µg/ml, 150 µg/ml, 200 µg/ml, 300 µg/ml, 350 µg/ml, 500 µg/ml, 650 µg/ml, etc.), as measured by gravimetric methods. In some embodiments, the total concentration of lipids and wax species in the bioactive propolis extract ranges from about 25 μg/ml to about 100 μg/ml, as measured by gravimetric methods. In some embodiments, the total concentration of lipids and wax species in the bioactive propolis extract ranges from about 75 μg/ml to about 250 μg/ml, as measured by gravimetric methods. In some embodiments, the total concentration of lipids and wax species in the bioactive propolis extract ranges from about 300 μg/ml to about 550 μg/ml, as measured by gravimetric methods.

[098]

Biologically active seaweed extracts

[099] In accordance with some embodiments, the composition contains a biologically active extract derived from seaweed. In some embodiments, the bioactive extract is a powder obtained by drying and grinding seaweed. Methods for obtaining seaweed extracts (eg powders) are known, for example, described in Costa et al. (2010) Biomedicine and Pharmacology 64:21–28.

[100] Seaweeds are generally classified into four families: Rhodonphyceae (red algae), Phaeophyceae (brown algae), Cyanophaeceae (blue-green algae), and Chlorophyceae (green algae). Identification and classification of seaweeds are described, for example, in AlgaeBase (Guiry, M.D. & Guiry, G.M. 2018. AlgaeBase. World-wide electronic publication, National University of Ireland, Galway; www.algaebase.org).

[101] In some embodiments, the bioactive seaweed extract comprises one or more types of red algae. Examples of red algae include, but are not limited to, Rhodophyta species (eg, Rhodophyta graciliara caudata, etc.), Porphyra species (eg, Porphyra haitanensis, etc.), Pterocladiella species (eg, Pterocladiella capillacea, etc.), Osmundaria (e.g. Osmundaria obtusiloba, etc.), Gelidium spp. (e.g. Gelidium cartilagenium, etc.), Chondrococcus spp. (e.g. Chondrococcus hornemannii, etc.) and Hypnea spp. (e.g. Hypnea musciformi, etc.) .d.). In accordance with some embodiments, the biologically active seaweed extract contains Porphyra haitanensis.

[102] In some embodiments, the bioactive seaweed extract comprises one or more types of brown algae. Examples of brown algae include, but are not limited to, Laminaria species (e.g., Laminaria japonica, etc.), Sargassum species (e.g., Sargassum wightii, Sargassum filipendula, etc.), Spatoglossum species (e.g., Spatoglossum schroderi, etc.) , Padina spp. (eg Padina tetrastromatica, etc.), Dictyota spp. (eg, Dictyota cervicornis, Dictyota menstrualis, Dictyota myrtensii, etc.), Dictyopteris spp. (eg, Dictyopteris delicatula, etc.). In accordance with some embodiments, the biologically active seaweed extract comprises Laminaria japonica.

[103] In some embodiments, the bioactive seaweed extract comprises one or more types of green algae. Examples of green algae include, but are not limited to, Ulva species (e.g., Ulva arasakii, Ulva armoricana, Ulva clathrata, Ulva conglobate, Ulvafasciata, Ulva pertusa, Ulva reticulate, Ulva rigida, Ulva rotundata, etc.), Enteromorpha species (e.g., Enteromorpha lima , Enteromorpha clathrata, Enteromorpha compressa, Enteromorpha intestinalis, Enteromorpha prolifera, etc.), Monostroma spp. (e.g. Monostroma latissimum, Monostroma nitidum, Monostroma angicava, etc.), Codium spp., Caulerpa spp. (e.g. Caulerpa brachyous, Caulerpa cupressoides, Caulerpa lentillifera, Caulerpaprolifera, Caulerpa racemosa, Caulerpa sertularioides, etc.), Bryopsis species (e.g. Bryopsisplumose, etc.), Elalimeda species (e.g. Elalimeda monile, etc.), Capsosiphon species ( e.g. Capsosiphon fulvescens, etc.) and Chaetomorpha species (e.g. Chaetomorpha antennenina, etc.). In accordance with some embodiments, the biologically active seaweed extract contains Ulva latuca. In accordance with some embodiments, the biologically active seaweed extract contains Enteromorpha linza. In accordance with some embodiments, the biologically active seaweed extract contains Ulva latuca and Enteromorpha linza.

[104] In some embodiments, the bioactive seaweed extract comprises one or more types of blue-green algae. Examples of blue-green algae include, but are not limited to, Microcystis spp. (e.g., Microcystis aeruginosa, etc.), Nostoc spp. (e.g., Nostoc linckia, Nostoc spongiaeform, etc.), Lyngbya spp. (e.g., Lyngbya majuscule, Lyngbya bouillonii, Lyngbya sordida, etc.), Symploca spp., Calothrix spp., etc.

[105] The biologically active extract may contain a combination of seaweed species, for example, at least one green seaweed species (for example, 2, 3, 4, 5 or more green seaweed species), at least one red seaweed species (for example, , 2, 3, 4, 5 or more species of red algae) and/or at least one species of brown algae (for example, 2, 3, 4, 5 or more species of brown algae). For example, the biologically active seaweed extract may contain the following species: Enteromorpha linza, Ulva lactula, Porphyra haitanensis and Laminaria japonica. In accordance with some embodiments, the seaweed extract does not contain (eg, does not have) one or more classes of seaweed species, such as red algae, green algae, brown algae, and blue-green algae. In accordance with some embodiments, the seaweed extract does not contain blue-green algae.

[106] The proportion of each type of seaweed in the mixture from which the biologically active seaweed extract is obtained can vary. For example, in some embodiments, the mixture contains green and brown seaweed in a ratio of about 5:1 to about 1:5 (e.g., 1:1, 1:2, 1:3, 1:4, 1:5 , 2:1, 2:3, 2:4, 2:5, 3:1, 3:2, 3:4, 3:5, 4:1, 4:2, 4:3, 4:5, 5 :1, 5:2, 5:3, 5:4, etc.). In some embodiments, the mixture contains green and red algae in a ratio of about 5:1 to about 1:5 (e.g., 1:1, 1:2, 1:3, 1:4, 1:5, 2: 1, 2:3, 2:4, 2:5, 3:1, 3:2, 3:4, 3:5, 4:1, 4:2, 4:3, 4:5, 5:1, 5:2, 5:3, 5:4, etc.). In some embodiments, the mixture contains kelp and red algae in a ratio of about 5:1 to about 1:5 (e.g., 1:1, 1:2, 1:3, 1:4, 1:5, 2: 1, 2:3, 2:4, 2:5, 3:1, 3:2, 3:4, 3:5, 4:1, 4:2, 4:3, 4:5, 5:1, 5:2, 5:3, 5:4, etc.). In some embodiments, the propolis mixture contains green, brown, and red algae in a ratio of about 5:1:1 to about 1:5:1 to about 1:1:5 (e.g., 1:1:1, 1 :2:1, 2:1:1, 1:1:2, etc.).

[107] The amount of biologically active seaweed extract in the composition of the present invention may vary. In accordance with some embodiments, the composition described herein (eg, a drug delivery composition) contains from about 0.1% wt./wt. up to approximately 20% w/w. biologically active seaweed extract. The amount of seaweed extract can be expressed as a percentage of the weight of each type of seaweed extract in the composition or as the total amount of biologically active seaweed extract in the composition. For example, the composition may contain 20% wt./wt. of the total amount of biologically active seaweed extract, of which the extract of each of the four different types of seaweed constitutes 5% w/w.

[108] In accordance with some embodiments, the composition contains from about 0.1% wt./wt. to approximately 5% w/w. biologically active seaweed extract. In accordance with some embodiments, the composition contains about 1%, about 2%, about 3%, about 4%, or about 5% wt./wt. biologically active seaweed extract. In accordance with some embodiments, the composition contains from about 3% to about 7% wt./wt. biologically active seaweed extract. In accordance with some embodiments, the composition contains from about 5% to about 20% w/w biologically active seaweed extract.

[109] Extracts derived from seaweeds (eg, one or more types of seaweed, eg a blend of two, three, four or more types of seaweed) may contain a number of biologically active agents. In accordance with some embodiments, the bioactive seaweed extract contains one or more of the following: sulfated polysaccharides, phorolotannins, xanthines (eg, fucoxanthin, astaxanthin, etc.), phloroglucinols, polyphenols, carotenoids, and sesquiterpenes.

[110] The present disclosure is based in part on bioactive seaweed extracts containing sulfated polysaccharides (eg, non-animal sulfated polysaccharides). Sulfated polysaccharides are anionic carbohydrate polymers that are usually classified based on the class of algae (eg, brown, green, red) from which they are derived. Typically, brown seaweeds (eg, Phaeophyceae) produce sulfated fucans or fucoidans, which may include fucose, xylose, uronic acid-based and/or galactose sugars. Red seaweeds (e.g., Rhodonphyceae) commonly produce galactans and carrageenans, which may include sulfated galactose and 3,6-anhydrogalactose sugars. Green seaweeds produce ulvans, which include sulfated rhamnose associated with either glucuronic acid, iduronic acid, or xylose. The classification of sulfated seaweed polysaccharides is known and, for example, described in Pater (2012) 3 Biotech 2(3): 171-185.

[111] The biological activity characteristics of sulfated polysaccharides may vary. In accordance with some embodiments, the biologically active seaweed extract contains one or more sulfated polysaccharides having antioxidant activity. Examples of antioxidant sulfated polysaccharides include, but are not limited to, ulvans extracted from Codia, Ulva and Enteromorpha, galactans extracted from Caulerpa, fucans and fucodans extracted from Laminaria japonicum, and galactans and carrageenans from Porphyra haitanensis. In accordance with some embodiments, the biologically active seaweed extract contains one or more sulfated polysaccharides having biological activity selected from anticoagulant activity, immunomodulatory activity, antitumor activity, antiviral activity, and antinociceptive activity. Methods for extracting and studying the functional characteristics of sulfated seaweed polysaccharides are known and described, for example, by Costa et al. (2010 Biomedicine and Pharmacotherapy 64:21-28; Zhang et al. (2010) Carbohydrate Polymers (2010) 82:118-121; and Wang et al. (2014) Mar. Drugs 12:4984-5020, full contents of each which are incorporated herein by reference.

[112] In some embodiments, the bioactive seaweed extract contains one or more sulfated polysaccharides selected from ulvan, carrageenan and fucan, or a combination of the above. In accordance with some embodiments, the biologically active extract contains one or more ulvans, for example, ulvans derived from Ulva latuca and/or Enteromorpha linza. In accordance with some embodiments, the biologically active extract contains one or more galactans and/or one or more carrageenans, for example, galactans and/or carrageenans derived from Porphyra haitanensis. In accordance with some embodiments, the biologically active extract contains one or more fucans and/or fucoidans, for example, one or more fucans and/or fucoidans derived from Laminaria japonica.

Biologically active extracts from carrots

[113] In accordance with some embodiments, the present invention relates to bioactive extracts containing combinations of molecules that are useful for promoting tissue regeneration (eg, a bioactive extract for tissue regeneration). In accordance with some embodiments, the bioactive extract for tissue regeneration contains one or more bioactive molecules derived from carrots (e.g., carotenoids, etc.), one or more vitamin A derivatives, one or more scaffold molecules, or a combination of any from the above.

[114] In accordance with some embodiments, one or more biologically active agents are derived from (eg, extracted from) carrots (eg, Dacus carota, or a cultivar or variant thereof). Carrots typically contain many bioactive molecules, such as carotenoids and phenolic compounds. Examples of carotenoids extracted from carrots include, but are not limited to, α-carotene and β-carotene. Examples of phenolic compounds extracted from carrots include chlorogenic acid, hydroxycinnamic acid derivatives, ferulic acid, dicaffeoylquinic acid and anthocyanins. In some embodiments, one or more bioactive molecules are extracted from the carrot in the form of “carrot oil,” which is typically obtained by either a solvent or pressure extraction process, such as cold pressing lipids and biomolecules from the seeds and/or taproot carrots; extracting compounds with a neutral oil (eg, mineral oil) from the seeds and/or taproot of carrots; alcohol extraction of biomolecules from carrot seeds and/or taproot; methods of supercritical extraction with carbon dioxide, etc. Methods for extracting biologically active molecules are also described, for example, in US Patent No. 7141083, US Publication No. 2008-0233238 and Japanese Patent No. JPH0676591.

[115] In some embodiments, the biologically active tissue regenerative extract comprises one or more B vitamin complexes (eg, one or more members of the B vitamin complex). A B vitamin complex generally refers to a complex containing all the essential water-soluble vitamins except vitamin C. In some embodiments, the B vitamin complex includes thiamine (vitamin B1), riboflavin (vitamin B2), niacin (vitamin B3), pantothenic acid (vitamin B5), pyridoxine (vitamin B6), biotin, folic acid and cobalamins (vitamin B12). In accordance with some embodiments, the bioactive extract for tissue regeneration contains pantothenic acid or a derivative, analogue or salt thereof. Examples of pantothenic acid derivatives and analogues include pantothenol and dexpanthenol. In accordance with some embodiments, the one or more B vitamin complexes are a pantothenic acid derivative or a derivative or analog thereof. In accordance with some embodiments, the vitamin B complex includes dexpanthenol.

[116] The total amount of biologically active carrot-derived extract in the composition may vary. According to some embodiments, the amount of the biologically active carrot-derived extract (e.g., carrot oil extract) ranges from about 1% (w/w) to about 5% (w/w), for example: about 1%, about 2%, about 3%, about 4% or about 5% w/w. In accordance with some embodiments, the bioactive extract for tissue regeneration does not contain (eg, does not have) the bioactive extract derived from carrots. In accordance with some embodiments, the biologically active extract for tissue regeneration further comprises Aloe vera, for example, Aloe vera extract.

[117] The present disclosure is based in part on bioactive extracts for tissue regeneration containing one or more scaffold molecules. As used herein, the term “scaffold molecule” refers to a molecule, such as a protein or polymer, that provides a scaffold to which cells (eg, stem cells, such as mesenchymal stem cells) can adhere and that promotes cell attachment, growth, and differentiation. In accordance with some embodiments, the one or more scaffold molecules are a protein (eg, an animal protein, an extracellular matrix (ECM) protein, etc.). In some embodiments, the protein includes collagen, albumin, or a combination thereof. In some embodiments, the protein comprises one or more whey proteins, such as albumin (eg, bovine serum albumin), fish whey proteins, porcine whey proteins, etc. In accordance with some embodiments, the composition contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more framework molecules.

[118] In some embodiments, the scaffold molecule is a polymer, such as a polysaccharide polymer, such as hyaluronic acid, chitosan, or alginate. In accordance with some embodiments, the polysaccharide polymer is hyaluronic acid. Hyaluronic acid is a glycosaminoglycan polymer that contains the repeating subunits of glucuronic acid and N-acetyl-D-glucosamine. Generally, hyaluronic acid is classified by its molecular weight. Low molecular weight hyaluronic acid (MWHA) typically has a molecular weight of less than 100 kDa. Medium weight MWHA typically has a molecular weight between 100 and 300 kDa. High mass MWHA typically has a molecular weight above 300 kDa. In accordance with some embodiments, the framework molecule comprises medium weight MWHA.

[119] The total number of scaffold molecules in the composition may vary. In some embodiments, the amount of scaffold molecules ranges from about 0.1% (w/w) to about 10% (w/w), for example: about 0.1%, about 0.5% , approximately 1%, approximately 2%, approximately 3%, approximately 4%, approximately 5%, approximately 6%, approximately 7%, approximately 8%, approximately 9%, approximately 10%, approximately 11%, approximately 12%, approximately 13%, approximately 14%, approximately 15%, approximately 16%, approximately 17%, approximately 18%, approximately 19% or approximately 20% w/w.

[120] The ratio of bioactive molecules to scaffold molecules in a bioactive extract for tissue regeneration can vary. For example, in accordance with some embodiments, the composition contains biologically active molecules and scaffold molecules in a ratio of from about 10:1 to about 1:10 (e.g., 1:1, 1:2, 1:3, 1:4, 1: 5, 1:6, 1:7, 1:8, 1:9, 1:10 2:1, 2:3, 2:4, 2:5, 2:6, 2:7, 2:8, 2 :9, 3:1, 3:2, 3:4, 3:5, 3:6, 3:7, 3:8, 3:9, 4:1, 4:2, 4:3, 4:5 , 4:6, 4:7, 4:8, 4:9, 5:1, 5:2, 5:3, 5:4, 5:6, 5:7, 5:8, 5:9, 6 :1, 6:2, 6:3, 6:4, 6:5, 6:7, 6:8, 6:9, 7:1, 7:2, 7:3, 7:4, 7:5 , 7:6, 7:8, 7:9, 8:1, 8:2, 8:3, 8:4, 8:5, 8:6, 8:7, 8:9, 9:8, 9 :7, 9:6, 9:5, 9:4, 9:3, 9:2, 9:1, 10:1, etc.).

Biologically active extracts from lemon balm

[121] In some aspects, the present invention relates to biologically active extracts obtained from Melissa officinalis, which is also called lemon balm. Melissa officinalis is a member of the mint family, Lamiaceae. Melissa officinalis is native to Europe and central Asia, but can generally be grown throughout the world. The varieties of M. officinalis include M. officinalis variety "citronella", M. officinalis variety "limonella", M. officinalis variety "Quedlinburger", M. officinalis variety "lime", M. officinalis variety "variegata", M. officinalis variety "aigea" iM. officinalis variety "Quedlinburger Niederliegende".

[122] Bioactive molecules produced by lemon balm include, but are not limited to, polyphenolic compounds, eugenol, tannins, and terpenes (e.g., monoterpenes, triterpenes, terpenoids, etc.), such as (+)-citronellal, 1-octen- 3-ol, 10-α-cadinol, 3-octanol, 3-octanone, α-cubebene, α-humulene, β-bourbonene, caffeic acid, caryophyllene, caryophyllene oxide, catechol, chlorogenic acid, cis-3-hexenol, cis -ocimene, citral A, citral B, copaene, δ-cadinene, eugenylacetate, γ-cadinene, geranial, geraniol, geranyl acetate, germacrene D, isogeranial, linalool, luteolin-7-glucoside, methylheptenone, neral, nerol, octyl benzoate, oleanolic acid , pomolic acid ((1R)-hydroxyursolic acid), protocatechuic acid, rhamnazine, rosmarinic acid, stachyose, succinic acid thymol, trans-ocimene, ursolic acid and harmine.

[123] In some embodiments, bioactive molecules are produced in the leaves and stems of lemon balm. Typically, bioactive molecules are isolated (eg, extracted) from lemon balm leaves in the form of an "essential oil", also called a "volatile oil". Methods for extracting biologically active compounds from lemon balm are known in the art, for example by aqueous extraction (eg as described in Nolkemper et al. (2006) Planta Med. 72(15): 1378-1382) or pressure extraction (eg as described in Dastmalchi et al. (2008) LWT - Food Science and Technology, 41 (3):391-400).

[124] In accordance with some embodiments, the biologically active extract obtained from Melissa officinalis contains one or more monoterpenes or monoterpenoids, for example, citronellal, neral and/or geranial. In accordance with some embodiments, the biologically active extract obtained from Melissa officinalis contains rosmarinic acid. In accordance with some embodiments, the terpenes and rosmarinic acid present in bioactive extracts of Melissa officinalis have antiviral (eg, antiretroviral, such as anti-herpes or anti-HIV) activity. The antiviral activity of biologically active extracts obtained from Melissa officinalis is described, for example, by Allahverdiyev et al. (2004) Phytomedicine 11(7-8):657-61, and Geuenich et al. (2008) Retrovirology 5:27.

[125] In some embodiments, the composition described herein contains a biologically active extract derived from Melissa officinalis and one or more antiviral agents, for example: acyclovir, valacyclovir, penciclovir, famciclovir, docosanol, avacavir, cidofovir, efavirenz, entecavir , imiquimod, lopinavir, emtricitabine, lamivudine, tenofovir, zidovudine, doravirine, etravine, nevirapine, rilpivirine, atazanavir, darunavir, fosamprenavir, sasquinavir, tipranivir, efuviritide, maraviroc, ralteravir, ibalzumab, cobicistat, or any combination thereof.

[126] In accordance with some embodiments, a composition comprising a bioactive extract obtained from Melissa officinalis further comprises one or more B vitamin complexes (eg, one or more members of the B vitamin complex). In some embodiments, one or more members of the B complex vitamins are cobalamins (eg, vitamin B12). In accordance with some embodiments, the cobalamins are methylated (eg, methylcobalamins) or scheduled (eg, cyanocobalamin). In accordance with some embodiments, the composition contains a combination of methylcobalamin and cyanocobalamin. Without wishing to be bound by any particular theory, in accordance with some embodiments, cobalamins are useful for promoting peripheral nerve repair and reducing neuropathic pain.

[127] The amount of cobalamin (eg, methylcobalamin and/or cyanocobalamin) in the composition can vary. According to some embodiments, the amount of methyl cobalamin in the composition ranges from about 0.1% (w/w) to about 0.5% (w/w), for example: about 0.1%, about 0.2%, approximately 0.3%, approximately 0.4% or approximately 0.5% w/w. According to some embodiments, the amount of cyanocobalamin in the composition ranges from about 0.1% (w/w) to about 2% (w/w), for example: about 0.1%, about 0.2 %, approximately 0.3%, approximately 0.4%, approximately 0.5%, approximately 0.6%, approximately 0.7%, approximately 0.8%, approximately 0.9%, approximately 1.0%, approximately 1.1%, approximately 1.2%, approximately 1.3%, approximately 1.4%, approximately 1.5%, approximately 1.6, approximately 1.7, approximately 1.8, approximately 1.9 or approximately 2.0% w/w

[128] The ratio of methylcobalamin to cyanocobalamin in the composition may vary. For example, in accordance with some embodiments, the composition contains methylcobalamin and cyanocobalamin in a ratio of from about 10:1 to about 1:10 (for example, 1:1, 1:2, 1:3, 1:4, 1:5, 1 :6, 1:7, 1:8, 1:9, 1:10 2:1, 2:3, 2:4, 2:5, 2:6, 2:7, 2:8, 2:9, 3:1, 3:2, 3:4, 3:5, 3:6, 3:7, 3:8, 3:9, 4:1, 4:2, 4:3, 4:5, 4: 6, 4:7, 4:8, 4:9, 5:1, 5:2, 5:3, 5:4, 5:6, 5:7, 5:8, 5:9, 6:1, 6:2, 6:3, 6:4, 6:5, 6:7, 6:8, 6:9, 7:1, 7:2, 7:3, 7:4, 7:5, 7: 6, 7:8, 7:9, 8:1, 8:2, 8:3, 8:4, 8:5, 8:6, 8:7, 8:9, 9:8, 9:7, 9:6, 9:5, 9:4, 9:3, 9:2, 9:1, 10:1, etc.).

Therapeutic methods

[129] In some aspects, the present invention relates to methods of delivering compositions (eg, therapeutic compositions) to a cell or subject. In accordance with some embodiments, the uses of the compositions described herein are useful for treating certain diseases and disorders associated with damaged tissue or damaged skin (e.g., diseased skin or cells), such as mucositis, ulcers, wounds (e.g., surgical incisions and etc.), burns and tissue damage due to viral infection (eg, peripheral nerve damage resulting from herpes virus 1 (HSV-1) infection or blisters resulting from herpes virus 2 (HSV-2) infection, chickenpox infection smallpox, etc.).

[130] The terms “treating,” “treating,” and “treating” refer to reversing, alleviating, delaying the onset of, or slowing the progression of a “pathological condition” described herein (e.g., a disease, disorder, or condition, or one or more features thereof or symptoms). In accordance with some embodiments, treatment may be administered after one or more signs or symptoms have appeared or been noticed. In other embodiments, treatment may be administered in the absence of signs or symptoms of a disease or condition. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (eg, in light of a history of symptoms and/or in light of genetic factors or other predisposition factors). Treatment may also be continued after symptoms resolve, for example, to delay or prevent recurrences of herpes virus infections.

[131] As disclosed herein, the compositions can be administered in any suitable manner. For example, an effective amount of the composition and/or other therapeutic agents can be administered to a subject in any manner that allows the agent to be delivered to the desired tissue, e.g., skin, mucosa, nervous system tissue, muscle tissue, etc. In accordance with some embodiments, the compositions are applied topically. Other suitable routes of administration include, but are not limited to, oral, parenteral, intravenous, intraperitoneal, intranasal, intramuscular, sublingual, intratracheal, inhalation, subcutaneous, ophthalmic, vaginal, and rectal. Systemic routes include oral and parenteral.

[132] When administered orally, the compositions may be administered in the form of tablets, pills, dragees, capsules, liquids, mouth rinses, gels, syrups, slurries, suspensions, etc. In accordance with some embodiments, the present invention provides compositions in in the form of a medicated mouthwash, spray, gel, cream, ointment, or dentifrice that, in some embodiments, eliminates bacterial biofilm responsible for infections and periodontal disease and promotes wound-healing effects on the lining of the cavity mouth, providing faster recovery of damaged tissue. In accordance with some embodiments, compositions for oral delivery contain a combination of a biologically active propolis extract and one or more surfactants (eg, one or more biological surfactants).

[133] In accordance with some embodiments, the composition formulated for oral delivery is compatible with the use of autologous transplantation techniques, for example, biomaterials such as platelet-rich fibrin (PRF). In accordance with some embodiments, the compositions of the present invention do not chemically react with grafted tissue and associated blood clots, making the compositions suitable for use on a subject undergoing dental procedures (eg, dental tissue transplantation, etc.).

[134] Therefore, in accordance with some embodiments, the compositions described herein are useful for treating mucositis (eg, oral lesions caused by radiation therapy, etc.). Mucositis is inflammation and ulceration of the oral mucosa that is usually caused by cancer chemotherapy and/or radiation therapy.

[135] In accordance with some embodiments, the present invention provides a method of treating oral lesions in a subject (eg, a subject with mucositis), the method comprising using a composition (eg, an oral care composition) as described herein by this disclosure, on a subject with one or more lesions in the oral cavity. In accordance with some embodiments, the oral care composition contains a biologically active extract derived from propolis. In accordance with some embodiments, the oral composition is administered in the form of a mouth rinse or an aqueous oral solution.

[136] In some embodiments, the subject having or suspected of having mucositis has already been subjected to chemotherapy (e.g., administration of one or more doses of a chemotherapeutic agent) and/or radiation therapy (e.g., exposure to one or more doses of therapeutic radiation eg radiation therapy to the head and neck). In accordance with some embodiments, the subject has previously been subjected to chemotherapy, radiation therapy, or a combination of chemotherapy and radiation therapy. In some embodiments, a subject having or suspected of having mucositis is subjected to whole body irradiation prior to receiving hematopoietic stem cells.

[137] In accordance with some embodiments, the compositions described herein (eg, oral compositions) are useful for treating tissue that has been mechanically damaged. In accordance with some embodiments, a subject with mechanically damaged oral tissue has undergone oral surgery, such as a dental implant procedure. In accordance with some embodiments, an oral care composition comprising a bioactive extract derived from propolis is compatible with biomaterials commonly used in oral surgery (eg, platelet-rich fibrin, etc.) and reduces the likelihood Compared to the use of alcohol-based compositions, such as antiseptic mouth rinses, the rejection of a dental implant by the subject's immune system.

[138] In accordance with some embodiments, use of an oral care composition described herein inhibits the formation and/or growth of a bacterial biofilm in a subject (eg, biofilm formation on the mucous membrane and/or in the oral cavity of the subject). Removal of bacterial biofilm from ulcerated mucosa results, in accordance with some embodiments, in a decrease in polymorphonuclear cell infiltration and a subsequent decrease in the release of proinflammatory mediators such as TNF-α, IL-1β, IL-6, and essential prostaglandins that mediate hyperalgesic pain.

[139] In some aspects, the present invention relates to methods for topical administration of the compositions described herein. Topical pharmaceutical formulations include transdermal patches, ointments, lotions, creams, gels, drops, sprays, suppositories, liquids, and powders. In addition, conventional pharmaceutical carriers, aqueous, powder or oily bases or thickeners can be used in topical pharmaceutical preparations. The compositions of the present invention, which are formulated for topical use, are useful, in accordance with some embodiments, for treating damaged tissue, such as skin. In accordance with some embodiments, the composition is a hydrogel. Typically, the term "hydrogel" refers to a network of polymer chains that are hydrophilic and form a colloidal gel in which water is the dispersion medium. In accordance with some embodiments, the hydrogel contains at least 10% wt./wt. water (for example, approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 99% water).

[140] "Injured tissue" (eg, damaged skin) refers to tissue that has been chemically or mechanically damaged. Examples of damaged tissue include wounds (eg, cuts, scrapes, puncture wounds, surgical incisions, neuropathic (diabetic) wounds, etc.), burns (eg, chemical burns, sunburn, heat burns, etc.) , blisters, ulcers, muscle and tendon tears, and degenerated nerve tissue.

[141] In some aspects, the present invention provides a method of treating damaged tissue in a subject, the method comprising topical application of a composition described herein to a subject in need. In accordance with some embodiments, the subject has an ulcer (eg, a diabetic ulcer). In accordance with some embodiments, the subject has a burn, such as a chemical burn (eg, as a result of a dermatological or cosmetic procedure, such as a chemical skin peel).

[142] In accordance with some embodiments, the composition is applied topically to a subject in the form of a spray, foam, gel, or aqueous solution. In accordance with some embodiments, the composition is applied to a subject as part of a device or apparatus (eg, a medical device), such as a solid support impregnated with or coated with the composition. Examples of solid substrates that can be impregnated or coated with the composition include, but are not limited to, fibers (eg, natural cotton fibers, synthetic fibers such as nylon, etc.), dressings, pads, coverings (eg, face masks), plastics , metals (for example, stainless steel, titanium, etc.), etc.

[143] In accordance with other embodiments, the composition described in the present disclosure is administered multiple times. In some cases, the composition can be applied daily, twice a week, weekly, every two weeks, every three weeks, monthly, every two months, every three months, every four months, every five months, every six months months or less than once every six months. In some cases, the composition is used several times a day, week, month and/or year. For example, the composition can be applied approximately every hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 12 hours, or more than twelve hours. In accordance with some embodiments, the composition is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10 times per day.

[144] Dosage regimens may be adjusted to achieve optimal therapeutic response. For example, the oligonucleotide can be administered intermittently, for example several doses per day, or the dose can be proportionally reduced depending on the needs of the therapeutic situation. One skilled in the art will readily be able to determine appropriate dosages and dosage schedules for the subject compositions, regardless of whether the compositions are to be administered to cells or subjects.

[145] Aspects of the present invention relate to methods of use in a subject (eg, a mammal). In accordance with some embodiments, the mammalian subject is a human or non-human primate. Non-limiting examples of non-human primate subjects include macaques (eg, cynomolgus or rhesus macaques), marmosets, tamarins, spider monkeys, three-lined monkeys, vervet monkeys, squirrel monkeys, baboons, gorillas, chimpanzees, and orangutans. In accordance with some embodiments, the subject is a human subject. Other examples of subjects include domestic animals such as dogs and cats; livestock such as horses, cattle, pigs, sheep, goats and chickens; and other animals such as mice, rats, guinea pigs and hamsters.

EXAMPLES

Example 1 Compositions for drug delivery

[146] This example describes several embodiments of the drug delivery compositions of the present invention. Typically, drug delivery compositions described herein contain the following components: a carbonate buffer solution, at least two different biological surfactants, at least one hygroscopic agent, at least one antioxidant and at least one biologically active means.

[147] The carbonate buffer system has a pH of from 7.5 to about 9.5, preferably from 8.0 to 9.0. When applied to damaged tissue of a subject, the drug delivery compositions described herein create a microenvironment that promotes tissue healing and regeneration. Certain ions present in buffers increase the concentration of large Na ⁺ or NH ₄ ⁺ ions in the composition or in the microenvironment created when the composition comes into contact with damaged tissue. Large ions bind to certain molecules on the surface of mammalian cells, such as heparin sulfate (HS), and prevent pathogens from entering cells by disrupting the binding of microorganisms such as bacteria and viruses to HS on the surface of target cells. The ability of the compositions described herein to create such an environment at such a pH is unexpected in light of currently available wound healing compositions, which typically have pH values in the neutral to acidic range.

[148] The combination of two biological surfactants in the compositions described herein has several uses. In accordance with some embodiments, biological surfactants disrupt the surface tension of cells, such as bacterial cells and cells of damaged tissue, thereby promoting molecular debridement of the wound. In addition, surfactants help the biologically active molecules of the composition penetrate deep into the damaged tissue.

[149] The hygroscopic agent provides moisture to the wound bed and promotes proper hydration of the tissue as it heals. In accordance with some embodiments, the hygroscopic agents also provide a matrix that can serve as a scaffold for cell adhesion and differentiation during tissue regeneration.

[150] Antioxidants present in the formulations scavenge free radicals and reduce reactive oxygen species (ROS), which is important in reducing inflammation in damaged tissue as the healing process progresses. Antioxidants can also play a role as preservatives in compositions.

[151] The compositions also include one or more biologically active agents that are delivered by the composition. As described in the examples below, the biologically active agent can be replaced or modified to include biologically active molecules that achieve the desired therapeutic result. Table 1 below shows one embodiment of the formulation described herein with a drug delivery composition.

[152] Thus, the drug delivery compositions disclosed herein create a microenvironment that promotes tissue healing and regeneration by simultaneously providing 1) molecular debridement of the wound, 2) proper hydration, and 3) anti-inflammatory and antimicrobial effects, such as removal of bacterial biofilm in resulting from disruption of the mucopolysaccharide framework structure or inhibition of pathogen binding to target cells of the subject.

Example 2. Composition for tissue restoration

[153] Currently, the main cornerstone of RIOM control is considered to be the reduction of symptoms and complications of RIOM, for example, through nutritional support, pain management, prevention and/or treatment of secondary infections.

[154] Regenerative therapy in dentistry involves the replacement and/or regeneration of oral tissues that have been altered by disease or injury. One of the mentioned aspects complicating this endeavor until now has been the complex nature of the tissues found in the oral cavity. These include both mineralized tissues, such as the cementum of teeth, alveolar bone and dentin, and soft tissues bound by ligaments (periodontal ligament), each of which contains separate populations of cells derived from different tissues (ectodermal and mesodermal).

[155] Platelet-rich fibrin (PRF) is obtained by simply centrifuging the patient's peripheral blood without anticoagulants and is therefore strictly autologous. This fibrin matrix contains platelets and white blood cells, as well as various growth factors and cytokines, including transforming growth factor-beta (TGF-β1), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), interleukin (IL)- lp, IL-4 and IL-6. In addition, fibrin formed during the final stages of the coagulation cascade, in combination with cytokines secreted by platelets, makes PRF a highly biocompatible matrix, especially in affected areas where the fibrin network also acts as a reservoir of tissue growth factors. Autologous PRF has been demonstrated to be more effective in oral tissue regeneration than any commercially available biomaterial in several randomized control trials (RCTs). However, traditional mouth rinses are designed to disinfect the mouth, facilitate the removal of bacterial plaque and prevent the formation of caries and are characterized by high concentrations of active compounds such as denaturing and antimicrobial agents. The use of mouth rinses containing ethanol, triclosan, and chlorhexidine are not recommended for rinsing postoperative PRF grafts because they may dissolve the blood clot and disrupt the fibrin membrane, affecting the healing process.

[156] This example describes a tissue repair solution useful for the prevention and healing of the ulcerative phase of RIOM, and as an adjuvant in dental implant procedures. As a rule, mouth rinse is used in the form of a solution for spraying directly on the affected areas of the mouth before and after meals, 3 times a day. Table 2 below describes an example tissue repair solution.

[157] The composition contains a composition for drug delivery, which is described in example 1, and includes biologically active substances from a balanced blend of high-quality propolis extract. In accordance with some embodiments, the composition exerts multiple simultaneous biological actions important for mucosal tissue repair, such as anti-inflammatory, antioxidant, immunological, antimicrobial, and antifungal actions. The composition is fully compatible with the use of autoplastic transplantation techniques, as in the case of PRF, since it does not have a chemical effect on the blood clot and the transplanted tissue.

[158] The biologically active propolis complex contains several groups of flavonoids, isoflavonoids, chalcones and pterocarpanes, which have antibacterial, antifungal and antioxidant effects important for wound healing and removal of bacterial biofilm from the lesion bed in the mucous and submucosal layers. The high antioxidant effect of isoflavonoid biologically active substances also promotes the removal of reactive oxygen species (ROS) resulting from DNA damage in irradiated cellular elements, which in turn causes a decrease in the destruction of the extracellular matrix of the oral mucosa.

[159] Removal of bacterial biofilm from ulcerated mucosa, in accordance with some embodiments, results in a decrease in polymorphonuclear cell infiltration and a subsequent decrease in the release of proinflammatory mediators such as TNF-α, IL-1β, IL-6, and essential prostaglandins , which mediate hyperalgesic pain.

[160] The presence of high-strength biomolecules such as rhamnolipids and sophorolipid biological surfactants in the drug delivery system promotes biochemical microdebridization, which leads to the effective destruction of the polysaccharide matrix of the bacterial biofilm and the removal of devitalized and necrotic tissues that appear due to cellular damage.

[161] All these synergistic actions together contribute to the rapid and effective restoration of tissue of the injured mucosa.

Example 3 Composition for wound healing

[162] Chronic wounds do not progress through the normal pattern of wound healing, but instead remain in a state of chronic inflammation, predominantly characterized by abundant infiltration of peripheral mononuclear cells (PMN) and macrophages (MF). The ever present inflammatory cells play an important role in the production of proinflammatory cytokines (eg, IL-1, TNF-α, IL-6) and the formation of a protease-rich and pro-oxidative hostile microenvironment. Increased proteolytic activity (eg, neutrophil elastase, MMP-8, and gelatinase) leads to the destruction of growth factors and extracellular matrix structural proteins critical for tissue repair. Increased levels of ROS (H ₂ O ₂ , O ₂ ^- ) may lead to direct damage to cells or extracellular matrix molecules or may promote increased expression of matrix metalloproteinases (MMPs) such as (MMP-1, -2, -3, -9 and -13). Bacterial biofilms affect chronic non-healing wounds in the following ways: 1) wounds that contain biofilms may remain undetected; 2) ineffective biofilm treatment may delay healing; 3) wound debridement is one of the most important treatment strategies for biofilms, but does not remove all biofilm and therefore cannot be used alone; 4) biofilms can quickly re-form; Periodic wound debridement alone is unlikely to prevent biofilm regrowth; however, effective topical antiseptic application within this time period can suppress biofilm re-formation; and 5) biofilms are present in most chronic wounds and can be located both on the surface and in the deeper layers of the wound. Biofilm components (eg, extracellular adhesion protein (Eap), formylmethionyl peptides, N-acetylmuramyl-L-alanyl-D-isoglutamine) may contribute to the disruption of host repair mechanisms by interfering with interactions with the cellular matrix or promoting the development of an inflammatory response.

[163] The following examples describe pharmaceutical compositions for delivery in the form of a cell-activating liquid foam that includes a bioactive extract containing sulfated seaweed polysaccharides with emollient and healing effects to stimulate granulation tissue growth, and a hydrogel that includes inhibitors of granulation tissue formation. bacterial biofilms such as propolis, baicalin and adrografolide to promote debridement and removal of dead tissue and foreign matter, and to maintain a moist environment conducive to wound healing. One embodiment of a liquid cell activating foam is described in Table 3A and one embodiment of a hydrogel for mitigating bacterial biofilm is described in Table 3B below.

[164] The biologically active components present in the composition act synergistically at various stages of the healing process of chronic wounds. For example, saponin compounds present in the Aloe vera phytocomplex, in combination with antibacterial lipid surfactants, promote the removal of bacterial biofilms by disrupting the mucopolysaccharide matrix, thereby reducing infection and the inflammatory response in the wound bed.

[165] Sulfated seaweed polysaccharides present in cell-activating liquid foam are important antioxidants and scavengers of free radicals such as hydroxy and superoxide (H ₂ O ₂ , O ₂ ), which reduce pro-oxidant and proteolytic activity, preventing the destruction of immunocytes, factors growth and extracellular matrix. Moreover, they can stimulate the immune system by controlling macrophage activity. Sulfated polysaccharides such as carrageenan, ulvan, and fucoidan can alter macrophage activity by increasing bacterial binding and cytolytic activity in a colonized wound. The anticoagulant and antithrombotic activity of sulfated polysaccharides also contributes to the synergistic effect of the product, preventing the formation of microthrombi and therefore promoting angiogenesis in new growing tissue.

Example 4. Cosmetic composition for skin restoration

[166] This example describes a therapeutic mask for facial skin restoration, impregnated with a composition suitable as a cosmeceutical agent in procedures performed after skin peeling, for example, pulsed light, chemical and fractional laser peels. The composition can also be used as a regenerative treatment after minor dermatological procedures to remove signs of disease, spots, cysts and freckles.

[167] The composition is a face mask made of biodegradable natural fibers that is impregnated or coated with a drug delivery composition containing biologically active substances such as beta-carotene (for example, derived from carrot oil), alpha-tocopherol and other components, important in the process of dermal regeneration. Biologically active substances are included in a liquid matrix of hyaluronic acid - dexpanthenol, which plays an important role in the preservation of remaining vegetative tissue, for example, after a laser burn.

[168] Hyaluronic acid (HA) also performs its function in the epidermis by maintaining the extracellular space and providing an open as well as hydrated structure for the passage of nutrients. Biologically active substances and nutrients such as retinoic acid (vitamin A), alpha-tocopherol (vitamin E) and vitamin B5 (dexpanthenol) can be actively transported along the HA matrix into the repaired tissue. In addition, NA likely plays a multifaceted role in mediating cellular and matrix events, with subsequent injury, inflammation, granulation tissue formation, reepithelialization, and remodeling. Table 4 below shows one example of a drug delivery composition described in this example.

Example 5. Therapeutic composition for herpes infection

[169] The HSV-1 virus resides in the skin of the lips, causing herpes simplex labialis (HSL). Herpes simplex labiali (HSL), also known as cold sore or cold sore, affects millions of Americans. It is estimated that there are 98 million cases of HSL each year in the United States alone.

[170] Initial infection with the virus occurs through direct contact between the mucous membranes or abraded skin of the lips or mouth and the saliva or other secretions of a person with an active primary or recurrent infection. Primary HSV infection usually occurs in early childhood and is often asymptomatic, but may also present as herpetic gingivostomatitis, which is characterized by the formation of vesicles (tiny blisters) and ulcers in the mouth and perioral area. Before HSL, warning signs appear that are known as "prodromal symptoms" - a feeling of pain, burning, itching or tingling at the site of subsequent development of blisters. During the prodromal stage, headache attacks may also occur. Within 24 hours of the prodromal stage, multiple grouped blisters appear, which then secrete a fluid phase until they form scabs. These scabs can often bleed quite easily, forming unsightly blackish scabs of dried blood that can bleed again when the skin is stretched, such as by eating and smiling. They usually heal without scars in 5-15 days. Herpes simplex can cause pain, discomfort, inconvenience and some psychological and social problems due to cosmetic disfigurement.

[171] After the initial infection, the virus remains in the sensory ganglia (nerve endings) in a latent form. Once reactivated, HSV migrates from these sensory ganglia to the outer layer of skin of the lips or mouth, causing relapse of HSL. The virus replicates in neurons, leading to recurrent outbreaks. Flares are often caused by exposure to ultraviolet light (sunlight and/or tanning beds), stress, immunosuppression, colds, fatigue, fever (hence the term “cold sore” or “cold sore”), overexposure to wind, extreme temperatures, menstrual cycle , pregnancy, dental procedure, or lip injury. A flare can be triggered by perioral laser resurfacing or injection of perioral botulinum toxin or filling materials. There is currently no cure for HSL, so in theory, once infected, the infection remains for life.

[172] This example describes a medicinal solution containing natural antiviral bioactive extracts in combination with peripheral nerve repair factors for the treatment of herpes simplex labialis (HSL) and genital herpes. The composition is a composition for topical use intended for the treatment of cold sores on the lip and genital herpes. There are currently no topical products available on the market that are designed to both reduce viral infection and repair damaged nerve tissue. Table 5 below describes one embodiment of such a composition for topical use.

[173] The biologically active extract of Melissa officinalis contains monoterpenoid compounds such as geranial, α-bisabolol, β-caryophyllene, linalool, neral, citronellal, α-cadinol, β-cadinene and others, which have synergistic antiviral effects against HSV-1 and HSV-2. In addition to monoterpenoid bioactives, bioactive extracts of Melissa officinalis contain polyphenolic compounds such as rosmarinic acid, which are characterized by antiviral activity against both herpes labialis (HSV-1) and genital herpes (HSV-2). In some embodiments, the composition further comprises or is used in conjunction with one or more antiviral agents (eg, acyclovir, etc.) that are currently used to treat HSV infection.

[174] Methylated vitamin B12 and methylcobalamin have been observed to be involved in the mechanism of repair of destroyed and damaged peripheral nerves, as well as in reducing neuropathic pain and stimulating nerve growth.

[175] Biological surfactants and ions present (via a buffer system) in the composition can, in some embodiments, block virus entry into target cells by preventing binding to HS on the cell surface.

Example 6: Evaluation of Hydrogel Formulations for Tissue Repair in an Ex Vivo Porcine Skin Model with Biofilm Formation

[176] This example describes experiments examining the effectiveness of four different wound debridements (formulations A, B, C and D) in a porcine dermal explant model. Briefly, 3-day-old mature explants were treated with gel formulations for 24 or 72 hours, followed by isolation of surviving microorganisms. Three representative species of microorganisms were used in this study: methicillin-resistant Staphylococcus aureus (Gram-positive), Pseudomonas aeruginosa (Gram-negative), and Candida albicans (yeast).

[177] Overall, tissue healing formulations tested showed good results at 72 hours post-treatment, indicating a significantly higher level of treatment efficacy compared to the 24 hour time point, with <1.5 log microbial load in all evaluated species.

[178] It is important to note here that the treatments were not reapplied, but instead the treatment after the initial treatment was left on the explants for the 72 hour treatment period.

Materials and methods

[179] Formulations Tested: Five different propolis-containing tissue healing formulations (Formulations A, B, C and D) were tested. Each composition was a hydrogel containing a different concentration of the active agent (for example, propolis). One hydrogel without active agent (vehicle) was tested as a negative control. Approximately 2 ml of each hydrogel formulation was applied to each explant used in the study.

[180] Test organisms: Tested for effectiveness against gram-positive and gram-negative bacteria, as well as fungal species. Table 6 lists the types and corresponding ATCC numbers.

Biofilm creation

[181] Porcine explant tissue was purchased from a United States Department of Agriculture (USDA) licensed facility that uses precision leveling technology to prepare porcine skin to a specific thickness of approximately 2 mm. Using a needle biopsy, the tissue was cut into circular explants of approximately 12 mm in diameter and artificially wounded using a Dremel tool, creating a wound approximately 2 mm in diameter with a cavity depth of 1.5 mm. The explants were then thoroughly washed and sterilized using chlorine gas. Before inoculation and application of test compounds, explants were placed on 0.5% agar in a 37°C incubator for approximately 2 hours to equilibrate. The explants were then inoculated with 15-20 μl of log-phase cultures of the indicated bacteria or a 48-hour culture of C. albicans at approximately 10 ⁵ CFU per explant and incubated for 3 days on 0.5% agar with daily transfer to fresh agar media .

Treatment

[182] 3-day-old explants were washed for 2 minutes in 2 ml of sterile PBS followed by treatment with the test compositions. Formulations A, B, C and D were loaded into 10 ml syringes to allow controlled application of approximately 2 ml of the compositions to the explants. After washing, the explants were transferred to individual wells of a 24-well plate, followed by the addition of test compounds. Then the plates were placed in a thermostat for 24 hours. In the second round of screening, treatment time was extended to 72 hours to assess whether there was benefit from increasing treatment duration. All four formulations and the control excipient of the dosage form were used in the same treatment. After the designated treatment time, the remaining microflora was isolated from all explants and enumerated using serial dilutions and culture.

Isolation of surviving bacteria

[183] After incubation for 24 or 72 hours, surviving bacteria and yeast were isolated from the tissue and counted. First, the remaining compounds were carefully removed from the explants by inverting the explants, followed by two washes in 2 ml of sterile PBS (2 minutes each) to remove any remaining materials. The washed explants were then placed in a 15-ml centrifuge tube containing 2 ml of Dey/Engley broth, a common neutralizer. Explants were vortexed for 10 s and subjected to a series of 5 sonication steps: 90 s sonication/60 s rest intervals. Samples were again vortexed to ensure homogeneity, serially diluted, and streaked onto medium (10 μl sample/spot, in triplicate) to a -6 dilution and streaked onto medium (200 μl undiluted isolation solution) if necessary.

Performance Data

[184] The data is summarized in Table 7 and FIG. 1-3.

[185] The effectiveness of tissue repair formulations against 3-day methicillin-resistant S. aureus biofilms is summarized in FIG. 1. Overall, formulations A-D showed comparable efficacy with approximately 5 log survival after 24 hours of treatment compared to approximately 7.5 log for untreated controls. Increased treatment time significantly reduced survival to levels below 1 log for Formulations A-C and approximately 1.5 log for Formulation D. Interestingly, a reduction in microbial load at the 24-hour treatment mark of almost 2 log was also observed for the vehicle. reduced but increased treatment time reduced S. aureus survival by only approximately 1 log, whereas significantly higher efficacy was observed for the formulations tested.

[186] The effectiveness of tissue repair formulations against 3-day P. aeruginosa biofilms is summarized in FIG. 2. Similar to what was reported for S. aureus, extended processing time increased the effectiveness of all formulations. For all formulations A-D, a reduction of more than 5 log was observed at 24 hours and less than 1 log in survival after 72 hours. These formulations appeared to be more effective against P. aeruginosa than S. aureus after 24 hours of treatment; however, vehicle alone also showed a time-dependent increase in efficacy against P. aeruginosa biofilms, with a treatment time of 72 hours resulting in 2 log survival.

[187] The effectiveness of tissue repair formulations against 3-day C. albicans biofilms is summarized in FIG. 3. The overall efficacy profile against C. albicans is similar to that observed for Gram-positive S. aureus, where for all formulations an increase in efficacy was observed with increasing treatment time (72 hours versus 24 hours treatment) and effectiveness at the 72 hour treatment time point was significantly higher than the effectiveness of the filler alone.

[188] Overall, all formulations evaluated in this study showed very high efficacy against biofilms of all three representative species of microorganisms evaluated: Gram-positive S. aureus, Gram-negative P. aeruginosa, and the yeast C. albicans. The extended treatment time of 72 hours resulted in an overall increase in potency for all formulations, resulting in <1.5 log viable bacteria for all microbial species. The excipient alone was also observed to reduce the microbial load by approximately 2-3 logs after 24 hours and 72 hours of treatment for S. aureus and C. albicans, and by 3 logs after 24 hours and almost 7 logs after 72 hours for P. aeruginosa.

[189] There are several important points to note here that may be relevant to explaining the observed effectiveness of the filler:

• in this study, the total microbial load was calculated, i.e. biofilm bacteria + planktonic bacteria. This may explain the partial effectiveness of the filler, especially in the case of 24 h treatment. For reference, if 3-day-old biofilm explants are treated for 24 hours in an antibiotic or antifungal solution at concentrations >200× MIC value, we will typically see a <2 log reduction in microbial load.

• P. aeruginosa produces biofilms with much higher biomass than other species. If vehicle components disrupt P. aeruginosa biofilm, this may result in an increase in the observed effectiveness, although this may not be due to killing of the microorganisms, but due to the physical removal of biofilms from the tissue.

Example 7: Evaluation of Wound Healing in a Porcine Penetrating Surgical Wound Model with Hydrogel Formulations Containing Biofilm Inhibitors

[190] This case study describes the evaluation of four hydrogel tissue healing formulations containing varying concentrations of propolis, baicalin, and andrographolide in a porcine model of penetrating skin surgical wound healing.

[191] The therapeutic properties of hydrogels for tissue repair (eg, hydrogels containing propolis) and three predicates for penetrating surgical wounds were studied in a porcine model. One male (great white) pig weighing approximately 39.5 kg was allowed to acclimatize for 3 days and was not given access to food for 12 hours before surgery. The animal was sedated by intramuscular ketamine (5 mg/kg), midazolam (0.5 mg/kg), and acepromazine (0.05 mg/kg). After sedation, the animal was anesthetized with an intravenous injection of propofol (5 mg/kg) with simultaneous inhalation of isoflurane to maintain sedation. An anesthetic block with 2% lidocaine was performed at the incision site. Before surgery, the dorsal surface was shaved and sterilized with 2% chlorhexidine. On the back of the pig, using a stainless steel template, a No. 11 scalpel and scissors, 16 through-and-through surgical wounds with a diameter of 2 cm were made symmetrically in four rows.

[192] Eight tissue healing treatments were tested in duplicate on these wound sites: InnovaCorium Dressing Hydrogel (IWD) - Formulation A, B, C and D, Plurogel® (PG - Medline, Northhill, IL), Medihoney ® (MH - Derma Sciences, Toronto, Canada), Amerigel® (AMG - Amerx, Clearwater, FL) and saline (SAL). For each treatment, sites were randomly selected according to FIG. 4. The amounts of selected components in formulations A, B, C and D are shown in Table 8 below.

[193] Each product in an amount of 0.5 g was administered to the wound bed every 24 hours during the 22 day study. The wounds were first washed with sterile saline solution to remove any product residue from previous use. The area around the wound was then wiped with sterile gauze to better secure the new dressing. To reduce stress and pain during dressing changes, the animal was sedated intramuscularly with ketamine (10 mg/kg) and midazolam (0.5 mg/kg) during the first week. The wound sites were covered with Curatec clear film (Urgo Medical, France) and 3M™ Micropore™ surgical tape (3M, Maplewood, MN). After treatment of the lesions, the animal was bandaged with a clean cotton bandage for greater comfort and protection from accidental injury.

[194] To evaluate wound healing efficiency, wounds were photographed every week to estimate area using Image J software (NIH, USA) with a circular template of known area as a reference. To assess the histological stage of wound healing, biopsies were taken from the wound bed on the 15th day (replication 1) and 22nd day (replication 2). Histological preparations were made with 3-μm sections and then left in an oven at 75°C for 30 minutes to drain the paraffin. Hematoxylin/eosin and Masson's trichrome staining was used. Briefly, samples were dewaxed in 3 xylene containers for 5 minutes each, then hydrated in 10 passes in decreasing concentrations of alcohol, i.e. 100%, 90%, 80%, 70%, and water. Nuclei were stained with hematoxylin according to Harris for 3 minutes, followed by washing in running water for 1 minute. Hematoxylin fixation was performed with 10 passes in Scoth solution and rinsing with running water for an additional 2 minutes. The cytoplasm was stained with eosin for 3 minutes. Finally, dehydration was carried out by increasing the alcohol content and clarifying in 3 xylene tanks with 10 passes each for assembly in a synthetic medium. Masson's trichrome staining was performed by dewaxing in 3 xylene containers for 5 minutes each and staining with hematoxylin for 3 minutes. Samples were fixed in Scoth solution in 10 passes. The samples were then treated with scarlet biebrich solution for 15 minutes, phosphotungstic acid/phospholybdenum for 10 minutes, aniline blue for 5 minutes, and acetic acid 1% for 2 minutes. Dehydration in alcohols of increasing concentration, clarification and assembly in a synthetic medium.

[195] In FIG. 5A-5H are photographs showing the healing process of penetrating surgical wounds on pig skin with various treatments and controls. Twenty-four postoperatively, the excisional wounds were clean with no signs of bleeding, infection, swelling, or erythema. After 7 days of treatment, all wounds treated with the tissue repair composition, regardless of whether it was Formulation A, B, C or D, showed slight impregnation of the product on the wound bed. There was some product impregnation in the area around the wound that was difficult to wash off with saline solution. These impregnations were more pronounced in the case of formulations with the highest concentration of propolis (Fig. 5A and 5D). Wounds treated with the various predicates and the saline control solution were also impregnated, but not as dark and thick as those in the propolis-containing formulations. After 14 days of treatment, there was no more accumulation of wound bed products on the affected areas. Among the formulations, formulation C had the largest granulation tissue area associated with the lowest percentage wound reduction (Figure 5C). One of the wounds treated with MN had hypergranulation and edema, but no infection, exudate or erythema was observed in any of the wounds during the entire study period (Fig. 5G). The data shows that in the first week of treatment, no significant reduction in lesion area was observed with any type of treatment compared to the control saline solution. However, in the second week of treatment, each formulation was observed to be more effective than the control saline solution. Table 9 shows the wound area for each treatment and control during the observation period. At the end of the study, Formulation A had the highest percentage wound reduction (98.6%) compared to the saline control (93.3%), as can be seen in FIG. 6. Treatment with MN did not result in wound healing during this study.

[196] The histological characteristics of intact porcine skin and all wounds treated with different products were evaluated. In biopsies taken from undamaged intact tissue, a very characteristic porcine dermal structure was observed with intact dense connective tissue with bundles of collagen fibers, the absence of an infiltrate of atypical and/or inflammatory cells and a small number of dermal papillae. In general, the observed fragments were completely epithelialized by a thin, uncompressed keratin layer. On histological sections of biopsies taken from wounds, it was observed that the epidermal structures were not as organized as intact skin, and were mainly characterized by the presence of inflammatory infiltrates and loose deposits of connective tissue with sparser and less dense fibers. On biopsies collected from wounds, it was observed that, in general, formulations A and D had greater tissue metabolism within 15 days with the formation of epidermis with intussusception and keratin deposition by stratified keratinocytes. In the case of composition C, this stage took the longest to reach. Among the predicates, AMG was one of those that restored epidermal and dermal structures the fastest. On the other hand, the MN product required the longest time to regenerate the skin layers. After 21 days, on histological sections of biopsy specimens of wounds treated with a composition for tissue restoration, complete organization and stratification of epidermal structures was observed with differences between different layers of the epidermis, even in lesions treated with composition C. On the other hand, by the end of 21 days of treatment, wounds treated through MN and SAL, were characterized by incomplete formation of the epidermis.

[197] From the data described above, it is clear that all tissue healing formulations (e.g., A, B, C, and D) were effective in healing penetrating excisional wounds in pigs compared to saline-treated lesions. The PG and AMG products also showed healing efficacy in the test. Composition A and AMG gave the best results, achieving a wound reduction percentage of over 98% in 21 days. To confirm tissue reorganization during skin repair, differences in the healing stages of excisional wounds in pigs between different treatments and controls were examined histologically. And this time again, advanced restructuring of the epidermal layer was observed in composition A, composition D and AMG after 15 days of treatment.

EQUIVALENTS

[198] Having thus described several aspects of at least one embodiment of the present invention, it will be appreciated that various changes, modifications and improvements will readily be made by those skilled in the art. Such changes, modifications and improvements are intended to form part of the present disclosure and fall within the spirit and scope of the present invention. Therefore, the above description and drawings are given by way of example only.

[199] Although several embodiments of the present invention have been described and illustrated herein, those skilled in the art will readily envision many other means and/or structures for performing functions and/or obtaining results and/or one or several of the advantages described herein, and each of such variations and/or each of such modifications are considered to fall within the scope of the present invention. More generally, those skilled in the art will readily understand that all parameters, dimensions, materials and configurations described herein are intended to be illustrative and that actual parameters, dimensions, materials and/or configurations will depend on the particular application or tasks for which the ideas of the present invention are applied. Those skilled in the art will understand or be able to ascertain by merely routine experimentation many equivalents for the specific embodiments of the present invention described herein. Accordingly, it is to be understood that the above embodiments are presented by way of example only, and that within the scope of the appended claims and their equivalents, the present invention may be practiced other than as specifically described and claimed herein. The present invention relates to each individual feature, system, article, material and/or method that is described herein. Furthermore, any combination of two or more such features, systems, products, materials and/or methods is included within the scope of the present invention, unless such features, systems, products, materials and/or methods are mutually incompatible.

[200] As used herein in the specification and claims, the singular number forms, unless expressly indicated otherwise, are to be understood to mean “at least one.”

[201] As used herein in the specification and claims, the phrase “and/or” should be understood to mean “either one or both” of the elements so connected, i.e. elements that in some cases are present together, and in other cases - separately. Other elements other than those specifically identified by “and/or” may be present, as appropriate, whether or not related to those elements specifically identified, unless expressly stated otherwise. Thus, as a non-limiting example, a reference to "A and/or B" when used in conjunction with open language such as "comprising" may in one embodiment refer to A without B (optionally including elements other than B); in another embodiment, to B without A (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.

[202] As used herein in the specification and claims, the phrase “or” is to be understood to have the same meaning as “and/or” as defined above. For example, when separating elements in a list, “or” or “and/or” should be interpreted as inclusive, i.e. including at least one, but also including more than one, of a number or list of elements, and, optionally, additional elements not specified in this list. Only terms clearly indicating the contrary, such as “only one of” or “exclusively one of” or, when used in a claim, “consisting of” will refer to the inclusion of exclusively one element from a number or list of elements. Generally, as used herein, the term “or” should be interpreted only as indicating exclusive alternatives (i.e., “one or the other, but not both”) when preceded by exclusivity terms such as “either one of,” “ one of", "only one of" or "exclusively one of". The phrase "substantially consisting of" when used in claims has its ordinary meaning as used in the field of patent law.

[203] As used herein in the specification and claims, the phrase "at least one" with respect to a list of one or more elements is to be understood to mean at least one element selected from any one or more elements in the list of elements, but not necessarily including at least one of each and every element specifically listed in the list of elements, and not excluding any combination of elements in the list of elements. This definition also allows that elements other than those specifically identified in the list of elements to which the phrase “at least one” refers may not necessarily be present, whether or not related to those specifically identified elements. Thus, by way of non-limiting example, “at least one of A and B” (or equivalently “at least one of A or B” or equivalently “at least one of A and/or B "") may refer in one embodiment to at least one, optionally including more than one, A, in the absence of B (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, in the absence of A (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one A, and at least one, optionally including more than one B (and optionally including other elements); etc.

[204] In the claims, as well as in the description above, all transitional phrases such as “comprising”, “including”, “carrying”, “having”, “containing”, “involving”, “holding”, etc. etc., should be understood as open, i.e. meaning inclusive without limitation. Only the transitional phrases “consisting of” and “substantially consisting of” must be closed or semi-closed transitional phrases, respectively, as specified in the US Patent Office Manual of Patent Examination Procedures, Section 2111.03.

[205] The use of ordinal terms such as “first”, “second”, “third”, etc., in claims to modify an element of a claim does not of itself imply any priority, advantage or order of priority element of a claim over another, or a temporary order in which the steps of a method are performed, but they are used only as markers to distinguish one element of a claim having a specific name from another element having the same name (but using an ordinal term ) to distinguish such elements of a claim.

[206] It should also be understood that, unless expressly stated otherwise, in any methods claimed herein that involve more than one step or more than one act, the order of the steps or steps of the method is not necessarily limited to the order in which the steps or steps are stated. of this method.

Claims (33)

1. Composition for regeneration and healing of skin tissue and mucous membranes, containing: (i) a drug delivery system comprising sodium bicarbonate, at least two biological surfactants selected from rhamnolipid and sophorolipid, at least one hygroscopic agent selected from polyethylene glycol polymer, propylene glycol polymer, hydroxyethylcellulose, hyaluronic acid, and at least one antioxidant selected from butylated hydroxytoluene (BHT) and alpha-tocopherol; And (ii) at least one biologically active agent selected from: a biologically active extract obtained from green propolis, a biologically active extract obtained from brown propolis, a biologically active extract obtained from Enteromorpha linza, a biologically active extract obtained from Ulva lactuca, biologically active extract obtained from Porphyra haitanensis, biologically active extract obtained from Laminaria japonica, Aloe vera, andrographolide, baicalin, biologically active extract obtained from carrot oil, biologically active extract obtained from Melissa officinalis, methylcobalamin, cyanocobalamin, dexpanthenol; And, wherein the composition has a pH in the range from about 7.5 to about 9.5. 2. The composition of claim 1, wherein the total amount of biological surfactants in the composition ranges from about 1.5% (w/w) to about 10% (w/w), and/or the amount hygroscopic agent in the composition ranges from about 1% (wt./wt.) to about 10% (wt./wt.). 3. The composition of claim 1 or 2, wherein the ratio of rhamnolipid to sophorolipid ranges from about 1:9 to about 9:1, optionally the ratio being about 1:1 or about 7:3. 4. The composition according to claim 1, wherein at least one biologically active agent is selected from a biologically active extract obtained from green propolis, or a biologically active extract obtained from brown propolis, or combinations thereof, optionally wherein the green propolis is Baccharis dracunculifolia propolis and optionally wherein the total amount of the bioactive extract obtained from green propolis and/or the bioactive extract obtained from brown propolis is in the range of from about 5% (w/w) to about 20% (w/w) .). 5. The composition of claim 1, wherein at least one biologically active agent is selected from a biologically active extract obtained from Enteromorpha linza, Ulva lactuca, Porphyra haitanensis and Laminaria japonica, and optionally wherein the total amount of biologically active extract is in the range of about 5 % (w/w) to approximately 20% (w/w). 6. The composition according to claim 1, wherein at least one biologically active agent is selected from a biologically active extract obtained from carrot oil; and/or at least one biologically active agent contains dexpanthenol; and/or the total amount of biologically active extract ranges from about 1% (w/w) to about 5% (w/w). 7. The composition according to claim 6, where the biologically active extract obtained from carrot oil contains one or more carotenoids and/or vitamin A. 8. The composition according to claim 6, wherein the composition further contains one or more proteins, wherein the protein includes collagen and/or albumin. 9. The composition according to claim 1, wherein at least one biologically active agent is selected from a biologically active extract obtained from Melissa officinalis, and optionally contains methylcobalamin and/or cyanocobalamin. 10. The composition according to claim 1, wherein at least one biologically active agent is selected from a biologically active extract obtained from Melissa officinalis and additionally contains dexpanthenol. 11. The composition of claim 8 or 9, wherein the total amount of biologically active extract ranges from about 1% (w/w) to about 5% (w/w). 12. Composition according to any one of paragraphs. 1-11, wherein the composition is formulated as a solid (eg, powder, such as lyophilized powder), liquid, gel or foam; optionally the liquid is a mouth rinse; gel or foam is formulated in the form of an aerosol spray or hydrogel; and/or the composition is presented on or in a solid support, which optionally contains cotton fibers and is optionally a bandage or fabric mask. 13. A method for treating lesions in the oral cavity of a subject, wherein the method involves using a composition according to any one of claims. 1-9 on a subject having one or more lesions in the oral cavity. 14. The method according to claim 13, wherein the subject: has mucositis or is suspected of having it; the subject has previously received chemotherapy, radiation therapy, or a combination of chemotherapy and radiation therapy; and/or underwent maxillofacial surgery, in particular a dental implantation procedure. 15. The method of claim 13 or 14, wherein the composition is applied directly to the oral cavity of the subject, optionally using a mouth spray or mouth rinse. 16. Method according to any one of paragraphs. 13-15, wherein the composition is administered to the subject more than once per day (eg, 2, 3, 4 or more times per day), optionally before meals, after meals, or both before and after meals. 17. Method according to any one of paragraphs. 13-16, and the use of the composition inhibits the formation and/or growth of bacterial biofilm. 18. A method for the regeneration and healing of skin tissue and mucous membranes, involving the use of a composition according to any one of paragraphs. 1-12 on the skin of the subject in need. 19. The method of claim 18, wherein the skin has been subjected to a cosmetic procedure, optionally wherein the cosmetic procedure is a laser skin peel, optionally wherein the subject's skin is damaged, optionally wherein the damaged skin is a wound, ulcer or blister. 20. The method of claim 18 or 19, wherein the use comprises bringing the subject's skin into contact with a solid support containing the composition, optionally the solid support being a non-adhesive bandage or cloth face mask. 21. The method according to claim 18 or 19, wherein the application is a topical application, which optionally occurs at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 times per day or per week. 22. Method according to any one of paragraphs. 18-21, and the subject: has or is suspected of having a chronic wound as a result of diabetes mellitus or damaged skin associated with insufficient blood flow (for example, varicose veins, bedsores, etc.); or is infected or suspected of being infected with herpes simplex virus, optionally the herpes simplex virus being herpes simplex labiali.