Use of Anti-microbial bioadhesive formulation

Abstract

A bioadhesive formulation comprises propylene glycol from 0.001 to 2 % w / w and a bioadhesive polymer from 50 to 99 % w / w. The bioadhesive formulation can be used in prevention or treatment of a microbial infection in a mucosal or dermal lesion. The bioadhesive formulation is mucoadhesive and / or dermoadhesive and is formulated for topical administration onto the mucosal or dermal lesion. The bioadhesive formulation inhibits microbial growth in the mucosal or dermal lesion.

Description

[0001] USE OF ANTI-MICROBIAL BIOADHESIVE FORMULATION

[0002] TECHNICAL FIELD

[0003] The present invention generally relates to prevention or treatment of a microbial infection in a mucosal or dermal lesion, and in particular to a bioadhesive formulation for use in prevention or treatment of a microbial infection in a mucosal or dermal lesion.

[0004] BACKGROUND

[0005] In the early phase of the wound healing process, wounds are often colonized with a variety of microbes, such as bacteria, fungi, parasites and / or viruses, which elicit host responses including hyperemia as well as specific cellular events, such as infiltration of leukocytes and inflammatory cells. Such inflammatory cells release cytokines, chemokines and other antimicrobial humoral factors, which together with the infiltrating cells initiate the proliferative and healing response leading to wound repair.

[0006] The granulation tissue in the wound cavity involves proliferation of fibroblasts, deposition of collagens and other extracellular matrices, and initiation of a development of new blood vessels. Once the infectious microbes are cleared, new tissue elements start to generate within the wound and the remodeling phase begins, aiming to restore tissue structural integrity and functional capacity in the healing wound. The different phases of the typical wound repair are usually not simple linear events but rather complex and overlapping in time during the healing process.

[0007] The development of new and effective interventions in wound care, especially to effectively initiate anti-infectious and anti-inflammatory processes, remains an area of intense research. The roles of various inflammatory cells, vasoactive amines, serotonin, the clotting factors, arachidonic acid metabolites, free radicals, platelet factors, cytokines, chemokines, and gene regulation are extensively studied, in terms of effects related to the inflammatory process. Despite several recent scientific discoveries and advances, the understanding of the mechanisms and pathways involved in normal and abnormal wound healing remain limited. Thus, there is significant room for improvements in the management of topical mucosal and / or skin wounds.

[0008] US 2022 / 0362166 discloses a pharmaceutical composition comprising from 0.001 to 2% (w / v) of propane-1, 2-diol as the only anti-inflammatory agent and at least one bioadhesive polymer. The pharmaceutical composition can be used in amelioration, prevention or treatment of an inflammatorycondition in a mucous membrane or skin of a subject, and / or in treatment of pain associated with such an inflammatory condition.

[0009] SUMMARY

[0010] It is a general objective to provide a treatment or prevention of a microbial infection in a mucosal or dermal lesion.

[0011] It is a particular objective to provide a bioadhesive formulation useful in the treatment or prophylaxis of a microbial infection in a mucosal or dermal lesion.

[0012] These and other objectives are met by embodiments as disclosed herein.

[0013] The present invention is defined in the independent claim. Further embodiments of the invention are defined in the dependent claims.

[0014] An aspect of the invention relates to a bioadhesive formulation for use in prevention or treatment of a microbial infection in a mucosal or dermal lesion. The bioadhesive formulation comprises propylene glycol from 0.001 to 2 % w / w and a bioadhesive polymer from 50 to 99 % w / w. The bioadhesive formulation is mucoadhesive and / or dermoadhesive and is formulated for topical administration onto the mucosal or dermal lesion. The bioadhesive formulation inhibits microbial growth in the mucosal or dermal lesion.

[0015] The bioadhesive formulation is formulated for topical administration onto a mucosal or dermal lesion and adheres to the mucosa membrane or skin containing the lesion. The bioadhesive formulation inhibits microbial growth at the lesion and has hygroscopic and absorptive properties that suppress any microbial infection and promote wound healing.

[0016] BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The embodiments, together with further objects and advantages thereof, may best be understood by making reference to the following description taken together with the accompanying drawings, in which:

[0018] Fig. 1 illustrates an immunohistochemically stained section of (A) bioadhesive patch, (B) superficial agar layer, and (C) deeper agar layer stained for IgG.Fig. 2 illustrates (1) a blood agar plate with inoculated Staphylococcus aureus with a bioadhesive patch positioned on the blood agar plate and (2) a 5 mm punch through the bioadhesive patch.

[0019] Fig. 3 illustrates S. aureus applied on top of a bioadhesive patch and incubated on a blood agar plate.

[0020] Fig. 4 illustrates tubes with inoculation from blood agar in Fig. 2 (tube 1), in Fig. 3 (tube 2) and positive control (tube 3).

[0021] Fig. 5 illustrates samples from tubes (1 )-(3) in Fig. 4 grown on mannitol-salt plates.

[0022] DETAILED DESCRIPTION

[0023] The present invention generally relates to prevention or treatment of a microbial infection in a mucosal or dermal lesion, and in particular to a bioadhesive formulation for use in prevention or treatment of a microbial infection in a mucosal or dermal lesion.

[0024] The bioadhesive formulation of the invention prevents, when topically administered onto a mucosal or dermal lesion, microbial growth in the lesion. Accordingly, the bioadhesive formulation is useful in preventing a microbial infection of such a mucosal or dermal lesion and finds use in treating an already ongoing microbial infection in a mucosal or dermal lesion.

[0025] The bioadhesive formulation is hygroscopic and absorptive and will thereby absorb liquid and molecules, including macromolecules, from a mucosal or dermal lesion when topically administered onto the lesion. It is believed that these properties of the bioadhesive formulation supports its prophylactic and therapeutic effects on microbial infections in mucosal and dermal lesions. In more detail, the hygroscopic properties of the bioadhesive formulation desiccate the lesion by absorbing inflammatory fluids, such as serous fluids and purulent fluids, from the wound, thereby reducing the risk of swelling at the lesion. Further, the absorptive properties of the bioadhesive formulation supports absorption of various molecules, including pro-inflammatory molecules, microbially released molecules and cellular components, from the lesion. In addition, the bioadhesive formulation may absorb microbes from the lesion, which thereby become removed from the lesion and trapped in the bioadhesive formulation. Taken together these effects of the bioadhesive formulation support healthy wound healing and create an antimicrobial environment or condition under the bioadhesive formulation that is unfavorable for microbial growth.Experimental data as presented herein shows that a bioadhesive formulation of the invention effectively prevents microbial growth, as represented by bacterial growth, when positioned on a blood agar plate inoculated with bacteria present in the skin and mucus microbiota. The bioadhesive formulation has also been shown herein to absorb large macromolecules as represented by immunoglobulin G (IgG), which are antibodies with a total molecular weight of about 150 kDa.

[0026] An aspect of the invention thereby relates to a bioadhesive formulation for use in prevention or treatment of a microbial infection in a mucosal or dermal lesion. The bioadhesive formulation comprises propylene glycol from 0.001 to 2 percent weight by weight (% w / w) and a bioadhesive polymer from 50 to 99 % w / w. The bioadhesive formulation is mucoadhesive and / or dermoadhesive and is formulated for topical administration onto the mucosal or dermal lesion. The bioadhesive formulation thereby inhibits microbial growth in the mucosal or dermal lesion.

[0027] % w / w as referred to herein, for instance, in the context of constituents of the bioadhesive formulation indicates the amount of a constituent (by weight) in 100 parts (by weight) of bioadhesive formulation.

[0028] Reference to an interval herein, such as from X to Y % w / w, encompasses the full range of the interval including the end points of the interval, i.e., X and Y, unless otherwise specified.

[0029] The bioadhesive formulation comprises propylene glycol, also referred to as propane-1, 2-diol, which is a synthetic diol alcohol with the chemical formula C3H8O2. The compound is sometimes called a-propylene glycol to distinguish it from the isomer propane-1, 3-diol, known as p-propylene glycol. Other alternative names include 1 ,2-propanediol, 1 ,2-dihydroxypropane, methyl ethyl glycol (MEG), and methylethylene glycol.

[0030] Polyethylene glycol is known for use in cosmetic and pharmaceutical compositions, as a solvent, cosolvent, humectant, stabilizer, and as a permeation enhancer. For a long time, propylene glycol has been considered as an inactive organic solvent (Seidenfeld and Hanzlik (1932) J. Pharmac. Exp. Ther. 44, 109-121; Braun and Cartland (1936) J. Am. Pharm. Ass. 25, 746-749; Weatherby and Haag (1938) J. Am. Pharm. Assoc. 27, 466-471). However, adverse effects of propylene glycol have occurred following topical, oral, and intravenous administration (Glascow, et al. (1983) Pediatrics 72(3), 353-355). The adverse effects associated with propylene glycol include central nervous system (CNS) toxicity, hyperosmolarity, hemolysis, cardiac arrhythmia, and lactic acidosis. Despite these adverse effects, highconcentrations (15-100%) have been advocated as to be used for topical administration on various skin lesions in order to kill bacteria, fungi, and / or virus (U.S. Patent No. 8,513,225). However, low concentrations of propylene glycol have never been shown to have any detrimental effects on living organisms.

[0031] Low concentrations of propylene glycol have positive effect on inflammatory conditions of mucous membranes and the skin by causing a significant reduction in the cytokines tumor necrosis factor alpha (TNF-a) and interleukin-2 (IL-2) (US 2022 / 0362166).

[0032] Propylene glycol is thereby included as an anti-inflammatory agent and in particular a pro-inflammatory cytokine suppressing agent that is capable of reducing TNF-a and IL-2 in the mucosal or dermal lesion.

[0033] The bioadhesive formulation comprises propylene glycol from 0.001 to 2 % w / w. In an embodiment, the bioadhesive formulation comprises propylene glycol from 0.01 to 1.5 % w / w. In another embodiment, the bioadhesive formulation comprises propylene glycol from 0.01 to 1 % w / w. In a further embodiment, the bioadhesive formulation comprises propylene glycol from 0.05 to 0.5 % w / w. In yet another embodiment, the bioadhesive formulation comprises propylene glycol from 0.075 to 0.25 % w / w. As an example, the bioadhesive formulation could comprise propylene glycol at about 0.1 % w / w.

[0034] The amounts of propylene glycol comprised in the bioadhesive formulation of the invention has medical effect but without the adverse effects seen with significantly higher concentrations of propylene glycol, such as disclosed in U.S. Patent No. 8,513,225.

[0035] The bioadhesive formulation further comprises a bioadhesive polymer. The bioadhesive polymer enables the bioadhesive formulation to adhere to the mucous membrane and / or skin to thereby be mucoadhesive and / or dermoadhesive. Accordingly, the bioadhesive polymer enables the bioadhesive formulation to cover a lesion in the mucous membrane or the skin.

[0036] The bioadhesive formulation comprises the bioadhesive polymer from 50 to 99 % w / w. The bioadhesive formulation can include one bioadhesive polymer or a mixture of multiple, i.e. , at least two, bioadhesive polymers. In the latter case, the total amount (by weight) of the multiple bioadhesive polymers in the bioadhesive formulation is from 50 to 99 % w / w.In an embodiment, the bioadhesive formulation comprises the bioadhesive polymer from 60 to 99 % w / w. In another embodiment, the bioadhesive formulation comprises the bioadhesive polymer from 70 to 99 % w / w. In a further embodiment, the bioadhesive formulation comprises the bioadhesive polymer from 80 to 99 % w / w. In yet another embodiment, the bioadhesive formulation comprises the bioadhesive polymer from 85 to 95 % w / w. As an example, the bioadhesive formulation could comprise the bioadhesive polymer at about 89.5 % w / w.

[0037] In an embodiment, the bioadhesive polymer is a mucoadhesive polymer. In such an embodiment, the bioadhesive formulation is preferably a mucoadhesive formulation formulated for topical administration onto a mucosal lesion. This means that the mucoadhesive polymer enables the mucoadhesive formulation to adhere to a mucous membrane of a subject. The mucoadhesive formulation then prevents microbial growth in the mucosal lesion.

[0038] In another embodiment, the bioadhesive polymer is a dermoadhesive polymer. In such an embodiment, the bioadhesive formulation is preferably a dermoadhesive formulation formulated for topical administration onto a dermal lesion. This means that the dermoadhesive polymer enables the dermoadhesive formulation to adhere to the skin of a subject. The dermoadhesive formulation then prevents microbial growth in the dermal lesion.

[0039] In a further embodiment, the bioadhesive polymer is a mucoadhesive and dermoadhesive polymer. In such an embodiment, the bioadhesive formulation is preferably a mucoadhesive and dermoadhesive formulation formulated for topical administration onto a mucosal lesion or a dermal lesion. This means that the mucoadhesive and dermoadhesive polymer enables the mucoadhesive and dermoadhesive formulation to adhere to a mucous membrane or the skin of a subject. The mucoadhesive and dermoadhesive formulation then prevents microbial growth in the mucosal lesion or the dermal lesion.

[0040] In an embodiment, the bioadhesive polymer is selected from the group consisting of a poly(acrylic acid) (PAA), an alginate polymer, carboxymethyl cellulose (CMC), polycarbophil, tragacanth, hydroxypropyl methylcellulose (HPMC), gum karaya, gelatin, guar gum, pectin, acacia, chitosan, hydroxypropyl cellulose, and any combination thereof.

[0041] Further examples of bioadhesive polymers that can be used according to the embodiments are disclosed in Kumar et al., (2014) Artificial Cells, Nanomedicine, and Biotechnology 42, 274-283.In a particular embodiment, the bioadhesive polymer is a PAA.

[0042] PAA, also referred to by its trade name carbomer, is a synthetic high-molecular weight polymer of acrylic acid. PAA may be homopolymers of acrylic acid, or crosslinked with an allyl ether of pentaerythritol, i.e., acrylic acid crosslinked with allyl pentaerythritol, an allyl ether of sucrose, i.e., acrylic acid crosslinked with allyl sucrose, or an allyl ether of propylene. PAA may also be in the form of copolymers of acrylic acid and C10-C30 alkyl acrylate crosslinked with allyl pentaerythritol. In a water solution at neutral pH, PAA is an anionic polymer, i.e., many of the side chains of PAA will lose their protons and acquire a negative charge. This makes PAAs polyelectrolytes, with the ability to absorb and retain water and swell to many times their original volume. In general, the carbomer resins that are intended for oral and mucosal applications are designated by a “P” (934P, 974P, 971 P).

[0043] PAA is swellable in water, due to a high percentage of carboxylic acid groups in the polymer. When dispersed in water, the PAA molecules partially swell, which increases viscosity. When neutralized with a water-soluble base, the PAA molecules swell completely, with a dramatic increase in viscosity. In an alkaline environment, the carboxyl groups of PAA dissociate; electrostatic repulsions between the negatively charged carboxyl groups cause uncoiling and expansion of the PAA molecule. This results in swelling of the polymer and gel formation. The gel is composed of closely packed swollen particles with swelling that increases with an increase in pH.

[0044] PAA is advantageous as bioadhesive polymer since it is physiologically inert and has good mucoadhesive and dermoadhesive properties. Furthermore, PAA may provide sustained release characteristics of the propylene glycol in the bioadhesive formulation to the mucosal or dermal lesion. Furthermore, PAA protects the underlying cell layer, forming a thick barrier that separates the lesion from the environment due to its continuous cross-linking and to the fact that it is water insoluble. For tablet dosage forms, PAA is a good disintegrant and a directly compressible vehicle. PAA produces tablets with excellent hardness and friability over a range of compression forces.

[0045] In an embodiment, the PAA is selected from the group consisting of a PAA having a viscosity from 29,400 to 39,400 cP at 25°C of a 0.5 % w / w aqueous solution at a pH from 7.3 up to 7.8 as determined by a Brookfield viscometer (Brookfield RVT, 20 rpm, spindle #6), a PAA having a viscosity from 4,000 to 11,000 cP at 25°C of a 0.5 % w / w aqueous solution at a pH from 7.3 up to 7.8 as determined by a Brookfield viscometer (Brookfield RVT, 20 rpm, spindle #5), and a combination thereof. Such PAAs aremarketed as CARBOPOL® 974P NF polymer, CARBOPOL® 971 NF polymer and CARBOPOL® 71 G NF polymer, respectively.

[0046] In a preferred embodiment, the PAA has a viscosity from 4,000 to 11,000 cP at 25°C of a 0.5 % w / w aqueous solution at a pH from 7.3 up to 7.8 as determined by a Brookfield viscometer. Such a PAA is marked as CARBOPOL® 71G NF polymer.

[0047] CARBOPOL® 71G NF polymer is a free-flowing granular form of CARBOPOL® 971 P NF polymer, for use in direct compression formulations. It is manufactured by roller compaction of CARBOPOL® 971 P NF polymer and is chemically the same product, with no additives. The resulting granules are free flowable, have increased bulk density, and contain minimal amounts of very small particles that can cause dusting and / or static adherence compared to the powder polymer.

[0048] In another embodiment, the PAA has a viscosity from 29,400 to 39,400 cP at 25°C of a 0.5 % w / w aqueous solution at a pH from 7.3 up to 7.8 as determined by a Brookfield viscometer. Such a PAA is marked as CARBOPOL® 974P NF polymer.

[0049] Other examples of bioadhesive polymers that can be used according to the invention include alginate polymers. Alginate, the salt of alginic acid, is a linear polysaccharide naturally produced by brown seaweeds (Phaeonphyceae, including Laminaria). Alginate is composed of multiple monomer residues, typically 100 to 3 000 monomers, linked together in a flexible chain. These residues are mainly of 0-(1 — >4)-linked D-mannuronic acid (M) residues and 0-(1 — >4)-linked L-guluronic acid (G) residues. These two residues are epimers and only differ at C5. In the polymer chain, they though give rise to very different conformations with any two D-mannuronic acid residues being4Ci-diequatorially linked while the link connecting any to L-guluronic acid residues is a1C -diaxial link as is illustrated in formula I:

[0050]

[0051] I

[0052] The residues are generally organized in blocks of identical or strictly alternating residues, e.g., MMMMMM..., GGGGGG..., or GMGMGM...

[0053] The alginate polymers form, in the presence of monovalent cations, dissolvable solid particles. This is in clear contrast to the case, when the alginate polymers are instead in contact with divalent cations, such as Ca2+. The divalent cations form links between different alginate polymers to thereby achieve crosslinking between the alginate polymers. The cross-linking in turn leads to the formation of an alginate film that is generally not dissolvable or at least difficult to dissolve in moist environments.

[0054] Suitable monovalent cations that can be used include sodium ions (Na+), potassium ions (K+) and ammonium ions (NH , preferably Na+.

[0055] The bioadhesive formulation may also comprise at least one excipient, such as at least one excipient selected from the group consisting of a diluent, a binder, a lubricant, an antiadherent, a coating, a colorant, a disintegrant, a flavor, a glidant, a preservative, a sorbent, a solvent, a buffer, a stabilizer, an adjuvant, a humectant, a surfactant, an osmotic agent, a sweetener, and any combination thereof.

[0056] In an embodiment, the bioadhesive formulation comprises at least one excipient selected from the group consisting of a diluent, a binder, a lubricant, and any combination thereof.

[0057] In an embodiment, the bioadhesive formulation comprises a diluent.

[0058] In an embodiment, the bioadhesive formulation comprises the diluent from 2.5 to 15 % w / w. In another embodiment, the bioadhesive formulation comprises the diluent from 2.5 to 12.5 %. In a further embodiment, the bioadhesive formulation comprises the diluent from 5 to 10 % w / w. In yet another embodiment, the bioadhesive formulation comprises the diluent from 6 to 9 % w / w. In a further embodiment, the bioadhesive formulation comprises the diluent from 7 to 9 % w / w. As an example, the bioadhesive formulation could comprise the diluent at about 7.9 % w / w.

[0059] In an embodiment, the diluent is selected from the group consisting of microcrystalline cellulose, lactose, starch, calcium phosphate, calcium carbonate, sucrose, maltodextrin, and any combination thereof.The bioadhesive formulation could comprise one diluent or a combination of multiple diluents.

[0060] In an embodiment, the diluent is microcrystalline cellulose. A currently preferred microcrystalline cellulose that can be used in the bioadhesive formulation has a bulk density from 0.26 to 0.31 g / mL.

[0061] In an embodiment, the bioadhesive formulation comprises a binder.

[0062] In an embodiment, the bioadhesive formulation comprises the binder from 0.01 to 5 % w / w. In another embodiment, the bioadhesive formulation comprises the binder from 0.1 to 5 % w / w. In a further embodiment, the bioadhesive formulation comprises the binder from 0.5 to 5 % w / w. In yet another embodiment, the bioadhesive formulation comprises the binder from 1 to 3 % w / w. As an example, the bioadhesive formulation could comprise the binder at about 2 % w / w.

[0063] In an embodiment, the binder is selected from the group consisting of povidone (polyvinyl pyrrolidone), starch, hydroxypropyl methylcellulose, microcrystalline cellulose, polyethylene glycol, and any combination thereof.

[0064] The bioadhesive formulation could comprise one binder or a combination of multiple binders.

[0065] In an embodiment, the binder is povidone.

[0066] In an embodiment, the povidone has a K value from 29.0 to 32.0.

[0067] In an embodiment, the povidone has a dynamic viscosity from 5.5 to 8.5 cP at 20°C of a 10 % w / v aqueous solution as determined by a rotational viscometer.

[0068] Povidone as used herein does not include povidone iodine. In an embodiment, povidone as used herein is 1-ethenylpyrrolidin-2-one.

[0069] In an embodiment, the bioadhesive formulation comprises a lubricant.

[0070] In an embodiment, the bioadhesive formulation comprises the lubricant from 0.01 to 5 %w / w. In another embodiment, the bioadhesive formulation comprises the lubricant from 0.01 to 2.5 % w / w. In a further embodiment, the bioadhesive formulation comprises the lubricant from 0.05 to 1 % w / w. In yet anotherembodiment, the bioadhesive formulation comprises the lubricant from 0.1 to 1 % w / w. In a further embodiment, the bioadhesive formulation comprises the lubricant from 0.25 to 0.75 % w / w. As an example, the bioadhesive formulation could comprise the lubricant at about 0.5 % w / w.

[0071] In an embodiment, the lubricant is selected from the group consisting of magnesium stearate, stearic acid, sodium stearyl fumarate, polyethylene glycol, glyceryl behenate, sodium lauryl sulfate, and any combination thereof.

[0072] The bioadhesive formulation could comprise one lubricant or a combination of multiple lubricants.

[0073] In an embodiment, the lubricant is magnesium stearate.

[0074] In an embodiment, the bioadhesive formulation comprises propylene glycol from 0.001 to 2 % w / w, the bioadhesive polymer from 50 to 99 % w / w, a diluent, preferably from 2.5 to 15 % w / w, a binder, preferably from 0.01 to 5 % w / w, and a lubricant, preferably from 0.01 to 5 % w / w.

[0075] In an embodiment, the bioadhesive formulation comprises, preferably consists of:

[0076] propylene glycol from 0.075 to 0.25 % w / w, preferably 0.1 % w / w;

[0077] poly(acrylic acid) from 85 to 95 % w / w, preferably 89.5 % w / w;

[0078] microcrystalline cellulose from 7 to 9 % w / w, preferably 7.9 % w / w;

[0079] povidone from 1 to 3 % w / w, preferably 2 % w / w; and

[0080] magnesium stearate from 0.25 to 0.75 % w / w, preferably 0.5 % w / w.

[0081] The bioadhesive formulation is formulated for topical administration onto a mucosal lesion or a dermal lesion. In an embodiment, the bioadhesive formulation is in a dosage form selected from the group consisting of a patch, a film, a tablet, a gel and a cream.

[0082] In an embodiment, the bioadhesive formulation is a bioadhesive tablet.

[0083] In another embodiment, the bioadhesive formulation is a bioadhesive patch.

[0084] For instance, a bioadhesive patch may be in the form of a dermal patch, also referred to as dermoadhesive patch, or a mucosal patch, also referred to as mucoadhesive patch, such as a mucoadhesive buccal patch. Such a bioadhesive patch may be shaped to cover the mucosal or dermallesion and, due to the presence of the bioadhesive polymer, may adhere to the lesion.

[0085] A bioadhesive patch can be manufactured according to various embodiments including in the form of a bioadhesive tablet, a bioadhesive film or a bioadhesive gel depending on the particular ingredients and concentrations of the ingredients.

[0086] The bioadhesive formulation of the embodiments is preferably for use in prevention or treatment of a microbial infection in the mucosal or dermal lesion caused by microbes selected from the group consisting of bacteria, fungi, viruses and parasites, and any combination thereof.

[0087] In an embodiment, the bioadhesive formulation of the embodiments is preferably for use in prevention or treatment of a microbial infection in the mucosal or dermal lesion. The bioadhesive formulation inhibits microbial growth in the mucosal or dermal lesion. The microbes are preferably selected from the group constating of bacteria, fungi, viruses and parasites, and any combination thereof.

[0088] In an embodiment, the bioadhesive formulation of the embodiments is preferably for use in prevention or treatment of a bacterial infection in the mucosal or dermal lesion caused by bacteria.

[0089] In an embodiment, the bacteria are selected from the group consisting of Staphylococcus bacteria, preferably Staphylococcus aureus, Streptococcus bacteria, preferably selected from the group consisting of Streptococcus sanguinis, Streptococcus mitis, Streptococcus pyogenes, Streptococcus epidermidis, Streptococcus mutans, Streptococcus salivarius, and any combination thereof, Prevotella bacteria, preferably Prevotella intermedia, Porphyromonas bacteria, preferably Porphyromonas gingivalis, Fusobacterium bacteria, preferably Fusobacterium nucleatum, Treponema bacteria, preferably Treponema denticola, Helicobacter pylori, and / or Pseudomonas aeruginosa.

[0090] In a particular embodiment, the bioadhesive formulation inhibits bacterial grow of Staphylococcus aureus, Streptococcus pyogene, Streptococcus epidermidis and / or Pseudomonas aeruginosa in the mucosal or dermal lesion.

[0091] In an embodiment, the bioadhesive formulation of the embodiments is preferably for use in prevention or treatment of a fungal infection in the mucosal or dermal lesion caused by fungi.In an embodiment, the fungi are selected from the group consisting of Actinomyces fungi, such as Actinomyces naeslundii, Actinomyces viscosus, and any combination thereof, Candida albicans, Aspergillus fumigatus, Coccidioides immitis, Histoplasma capsulatum, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Geotrichum fungi, Fusarium fungi, Rhodotorula fungi, Saccharomyces fungi and / or Penicillium mameffei.

[0092] In an embodiment, the bioadhesive formulation of the embodiments is preferably for use in prevention or treatment of a viral infection in the mucosal or dermal lesion caused by viruses.

[0093] In an embodiment, the viruses are selected from the group consisting of Herpes simplex viruses 1 , (HSV-1), HSV-2, Herpes zoster virus, Epstein-Barr virus (EBV), and / or Cytomegalovirus (CMV).

[0094] In an embodiment, the bioadhesive formulation of the embodiments is preferably for use in prevention or treatment of a parasite infection in the mucosal or dermal lesion caused by parasites.

[0095] In an embodiment, the parasites are selected from the group consisting of parasites, such as Trichomonas tenax and / or Entamoeba gingivalis, in the mucosal or dermal lesion.

[0096] The bioadhesive formulation is preferably used in prevention or treatment of a microbial infection in a mucosal or dermal lesion in a mammalian subject. The mammalian subject is preferably a human subject. The bioadhesive formulation may, however, also be applied in veterinary applications, where the subject is a non-human mammal, such as cat, dog, horse, sheep, pig, cow, goat, rabbit, guinea pig, rat, mouse, etc.

[0097] In an embodiment, the bioadhesive formulation does not comprise any active agent or ingredient, in particular no active agent or ingredient having anti-inflammatory effect, in addition to propylene glycol. Hence, any additional ingredients of the bioadhesive formulation are the bioadhesive polymer and one or more excipients. These one or more excipients and the bioadhesive polymer are not regarded as being active agents or ingredients in the bioadhesive formulation.

[0098] Another aspect of the invention relates to use of the bioadhesive formulation in the manufacture of a medicament for prevention or treatment of a microbial infection in a mucosal or dermal lesion.

[0099] A further aspect of the invention relates to a method for preventing or treating a microbial infection in amucosal or dermal lesion. The method comprises topically administering the bioadhesive formulation onto the mucosal or dermal lesion, wherein the bioadhesive formulation inhibits microbial growth in the mucosal or dermal lesion.

[0100] Treating or treatment as used herein and is well understood in the art, means an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results could include, for instance, alleviation or amelioration of one or more symptoms or conditions associated with the microbial infection, diminishment of extent of the microbial infection, stabilized state of the microbial infection, i.e., prevent worsening, preventing spread of the microbial infection, delay or slowing of progression of the microbial infection, amelioration or palliation of the state of the microbial infection, diminishment of the reoccurrence of the microbial infection, and remission.

[0101] Preventing, prevention or prophylaxis as used herein and is well understood in the art, means an approach in which a risk of developing a microbial infection in a mucosal or dermal is reduced or prevented, including prolonging or delaying development of any microbial infection in a mucosal or dermal lesion.

[0102] The various embodiments of the bioadhesive formulation described in the foregoing also apply to the method for preventing or treating a microbial infection in mucosal or dermal lesion. In particular, the various concentration intervals and values and ingredients of the bioadhesive formulation also apply to the method.

[0103] The bioadhesive formulation is anti-inflammatory and thereby inhibits or suppresses any inflammation at the mucosal or dermal lesion. This anti-inflammatory effect is partly due to presence of propylene glycol, which inhibits the pro-inflammatory cytokines TNF-a and IL-2. The bioadhesive formulation furthermore, due to its hygroscopic properties, absorbs inflammatory fluids from the lesion and, due to its bioabsorptive properties, absorbs pro-inflammatory molecules, microbially-released molecules and cellular components from the lesion. The bioadhesive formulation thereby cleanses and detoxifies the mucosal or dermal lesion, which promotes a healthy wound healing process.

[0104] A significant advantage of the bioadhesive formulation is that it has dual effects, i.e., both anti-microbial and anti-inflammatory effects. Such dual effects are in particular advantageous for the treatment of mucosal or dermal lesions caused by microbial infections. In such an application, the bioadhesiveformulation both treats the actual microbial infection due to is anti-microbial effects but also treat any inflammation induced by the microbial infection at the mucosal or dermal lesion.

[0105] The bioadhesive formulation may therefore be regarded as a hygroscopic bioadhesive formulation capable of absorbing fluids. In an embodiment, the bioadhesive formulation is capable of absorbing fluids corresponding to at least 10 times a weight of the bioadhesive formulation, preferably at least 15 times the weight of the bioadhesive formulation, more preferably 20 times the weight of the bioadhesive formulation, and most preferably 25 times the weight of the bioadhesive formulation.

[0106] The bioabsorptive properties of the bioadhesive formulation may even allow the bioadhesive formulation to absorb, and thereby extract or remove, microbes from the mucosal or dermal lesion. The bioadhesive formulation may thereby extract not only harmful or potentially harmful molecules from the lesion but also remove microbes from the lesion and capture them in the bioadhesive formulation.

[0107] The hygroscopic and bioabsorptive properties of the bioadhesive formulation further create an antimicrobial environment in the mucosal or dermal lesion, which suppresses microbial growth in the lesion.

[0108] Taken together these effects of the bioadhesive formulation thereby prevent or treat the microbial infection in the mucosal or dermal lesion.

[0109] EXAMPLES EXAMPLE 1

[0110] This example discloses the manufacturing of a bioadhesive patch. The ingredients of the bioadhesive patch are presented in Table 1.

[0111] Trade name Pharmacopoeia Europaea name Amount (% w / w) VIVAPUR® 101 Microcrystalline cellulose 7.9 VIVAPHARM® PVP K30 Povidone 2.0 Propylene glycol Propylene glycol 0.1 CARBOPOL® 71 G NF polymer Carbomer 89.5 LIGAMED® MF-2-V Magnesium stearate 0.5

[0112] Total 100.0

[0113]

[0114] A 5 % w / w aqueous solution of povidone with propylene glycol was prepared by mixing 0.25 g of povidone in 4.75 g purified water while stirring on a magnetic stirrer until povidone dissolved completely. 5.00 g of propylene glycol was added and mixed with the povidone solution.

[0115] The 5 % w / w aqueous solution of povidone with propylene glycol was used to wet 20.00 g of microcrystalline cellulose in a ceramic mortar. Granules were formed by wet sieving using a 0.6 mm sieve followed by vacuum drying in a temperature of 50°C to reduce the water content of the granules below 3 % w / w (as determined by Karl-Fisher titration using HYDRA-POINT™ composite 5 as titrant) and then dry sieving using a 0.5 mm sieve.

[0116] 12.625 g of the granules were mixed with 113.0 g of the carbomer for 5 min followed by addition of 0.631 g of magnesium stearate and mixing for 3 min.

[0117] Bioadhesive patches having a diameter of 9 mm, a thickness of 1.35 mm and a weight of 80 mg were produced using a laboratory rotary tablet press (Erweka RTP-D8, Germany) equipped with 9 mm flat, standard EU “D” punches and dies (Natoli Engineering Company, U.S.) with a rotor speed of 10 rpm, a feeder speed of 30 rpm and a main compression force of 4.9 kN.

[0118] EXAMPLE 2

[0119] This example investigated the antibacterial effects of a bioadhesive patch manufactured in Example 1.

[0120] A broth of Streptococcus epidermidis was applied with an inoculating loop over a blood agar plate with a diameter of 9 cm. A bioadhesive patch as manufactured in Example 1 was placed on the agar plate. The agar plate was incubated at 37°C for 24 hours. Following the incubation, the bioadhesive patch was carefully removed from the agar plate. The surface previously covered by the bioadhesive patch was sampled for bacteria by the inoculating loop and applied on a new blood agar plate. This agar plate was incubated at 37°C for 72 hours, and examined by an experienced medical laboratory scientist. No growth of S. epidermidis was observed on the agar plate following 72 hours of incubation.

[0121] The bioadhesive patch stopped bacterial growth on the blood agar surface covered by the bioadhesive patch. The bioadhesive patch further prevented ingrowth of bacteria into the surface covered by the bioadesive patch.EXAMPLE 3

[0122] This example investigated the absorption properties of a bioadhesive patch as manufactured in Example 1.

[0123] A 3 mm well was punched in a blood agar plate with a diameter of 9 cm. The well was filled with human serum and a bioadhesive patch manufactured in Example 1 was applied over the well. The agar plate was incubated at 37°C for 24 hours. The entire bioadhesive patch together with the agar including the 3 mm well was removed by a scalpel and transferred to a piece of cork and then covered with freeze medium. The specimen was then submerged in liquid nitrogen and stored at -20°C.

[0124] Seven m thick sections were obtained through sectioning of the frozen specimen in a cryostat. The sections were stained for immunoglobulin G (IgG) using a rabbit recombinant monoclonal human IgG antibody (ab109489; Abeam, UK) as the primary antibody. Staining for IgG was visualized using a DAB chromogen kit (Biocare Medical, USA). All sections were counterstained by Mayers HTX (Histolab Products AB, Sweden).

[0125] The immunohistochemical staining verified absorption of the IgG macromolecules by the bioadhesive patch as shown in Fig. 1. In Fig. 1, A represents the bioadhesive patch with positive staining for IgG, B represents the superficial layer of agar where IgG staining was absent, and C represents the deeper layer of the agar where staining for IgG was observed.

[0126] It was concluded that the bioadhesive patch had absorbed IgG from the superficial layer of the 3 mm well initially filled with human serum.

[0127] EXAMPLE 4

[0128] This example investigated whether a bioadhesive patch manufactured in Example 1 could eliminate bacteria. The first part of the example studied the ability of the bioadhesive patch to mortify Staphylococcus aureus. The second part of the example studied the ability of the bioadhesive patch to prevent growth of S. aureus when applied on the surface of the bioadhesive patch.

[0129] A broth of S. aureus was applied with an inoculating loop over a blood agar plate with a diameter of 9 cm. A bioadhesive patch as manufactured in Example 1 was placed on the agar plate. The agar plate was incubated at 37°C for 24 hours (Fig. 2: 1). Following the incubation, a 5 mm punch was used throughthe bioadhesive patch (Fig. 2: 2). The punched cylinder was transferred to a broth and incubated for another at 37°C for 24 hours.

[0130] In the second part, S. aureus was applied with a sterile loop on top of a bioadhesive patch as manufactured in Example 1 and incubated at 37°C for 24 hours (Fig. 3).

[0131] Tubes with the broth from the first and second part (Fig. 4: tubes 1-2) were examined for growth. A positive control for growth of S. aureus was also carried out (Fig. 3). In addition, samples from the tubes (Fig. 4: tubes 1-3) were grown on mannitol-salt plates (Fig. 5: plates 1-3).

[0132] As can be seen in Figs. 2 and 4, the bioadhesive patch killed the bacteria covered by the patch (Figs. 2, 4: tube 1). A peripheral zone around the patch also lacked bacterial growth (Fig. 2: 1).

[0133] The second part of this example revealed that the bioadhesive patch prevented growth of S. aureus when applied on the top of bioadhesive patch (Figs. 3, 4: tube 2). This example supports that the bioadhesive patch is mortifying S. aureus.

[0134] The results presented in Fig. 5: plates 1-2 confirm no growth in the broth-containing tubes (Fig. 4: tubes 1 -2). Fig. 5: plate 3 is a positive control comprising the sample in tube 3 in Fig. 4.

[0135] The bioadhesive patch was concluded to have the capacity to mortify and prevent growth of S. aureus.

[0136] The embodiments described above are to be understood as a few illustrative examples of the present invention. It will be understood by those skilled in the art that various modifications, combinations and changes may be made to the embodiments without departing from the scope of the present invention. In particular, different part solutions in the different embodiments can be combined in other configurations, where technically possible. The scope of the present invention is, however, defined by the appended claims.

Claims

CLAIMS1. A bioadhesive formulation for use in prevention or treatment of a microbial infection in a mucosal or dermal lesion, whereinthe bioadhesive formulation comprises:propylene glycol from 0.001 to 2 % w / w; anda bioadhesive polymer from 50 to 99 % w / w;the bioadhesive formulation is mucoadhesive and / or dermoadhesive and is formulated for topical administration onto the mucosal or dermal lesion; andthe bioadhesive formulation inhibits microbial growth in the mucosal or dermal lesion.

2. The bioadhesive formulation for use according to claim 1, wherein the bioadhesive formulation comprises propylene glycol from 0.01 to 1.5 % w / w, preferably from 0.01 to 1 % w / w, more preferably from 0.05 to 0.5 % w / w, and most preferably from 0.075 to 0.25 % w / w, such as 0.1 % w / w.

3. The bioadhesive formulation for use according to claim 1 or 2, wherein the bioadhesive formulation comprises the bioadhesive polymer from 60 to 99 % w / w, preferably from 70 to 99 % w / w, more preferably from 80 to 99 % w / w, and most preferably from 85 to 95 % w / w, such as 89.5 % w / w.

4. The bioadhesive formulation for use according to any one of claims 1 to 3, the bioadhesive polymer is a polyfacrylic acid).

5. The bioadhesive formulation for use according to claim 4, wherein the poly(acrylic acid) has a viscosity from 4,000 to 11 ,000 cP at 25°C of a 0.5 % w / w aqueous solution at pH 7.3 to 7.8 as determined by a Bookfield viscometer.

6. The bioadhesive formulation for use according to any one of claims 1 to 5, further comprising a diluent from 2.5 to 15 % w / w.

7. The bioadhesive formulation for use according to claim 6, wherein the bioadhesive formulation comprises the diluent from 2.5 to 12.5 % w / w, preferably from 5 to 10 % w / w, more preferably 6 to 9 % w / w, and most preferably from 7 to 9 % w / w, such as 7.9 % w / w.

8. The bioadhesive formulation for use according to claim 6 or 7, wherein the diluent is microcrystalline cellulose.

9. The bioadhesive formulation for use according to claim 8, wherein the microcrystalline cellulose has a bulk density from 0.26 to 0.31 g / mL.

10. The bioadhesive formulation for use according to any one of claims 1 to 9, further comprising a binder from 0.01 to 5 % w / w.

11. The bioadhesive formulation for use according to claim 10, wherein the bioadhesive formulation comprises the binder from 0.1 to 5 % w / w, preferably from 0.5 to 5 % w / w, more preferably from 1 to 3 % w / w, and most preferably 2 % w / w.

12. The bioadhesive formulation for use according to claim 10 or 11 , wherein the binder is povidone.

13. The bioadhesive formulation for use according to claim 12, wherein povidone has a K value from 29.0 to 32.0.

14. The bioadhesive formulation for use according to claim 12 or 13, wherein povidone has a dynamic viscosity from 5.5 to 8.5 cP at 20°C of a 10 % w / v aqueous solution as determined by a rotational viscometer.

15. The bioadhesive formulation for use according to any one of claims 1 to 14, further comprising a lubricant from 0.01 to 5 % w / w.

16. The bioadhesive formulation for use according to claim 15, wherein the bioadhesive formulation comprises the lubricant from 0.01 to 2.5 % w / w, preferably from 0.05 to 1 % w / w, more preferably from 0.1 to 1 % w / w, and most preferably from 0.25 to 0.75 % w / w, such as 0.5 % w / w.

17. The bioadhesive formulation for use according to claim 15 or 16, wherein the lubricant is magnesium stearate.

18. The bioadhesive formulation for use according to any one of claims 1 to 17, wherein the bioadhesive formulation comprises, preferably consists of:propylene glycol from 0.075 to 0.25 % w / w, preferably 0.1 % w / w;poly(acrylic acid) from 85 to 95 % w / w, preferably 89.5 % w / w;microcrystalline cellulose from 7 to 9 % w / w, preferably 7.9 % w / w;povidone from 1 to 3 % w / w, preferably 2 % w / w; andmagnesium stearate from 0.25 to 0.75 % w / w, preferably 0.5 % w / w.

19. The bioadhesive formulation for use according to any one of claims 1 to 18, wherein the bioadhesive formulation is a selected from the group consisting of a bioadhesive patch, a bioadhesive tablet, a bioadhesive cream and a bioadhesive gel, preferably selected from the group consisting of a bioadhesive patch and a bioadhesive tablet, more preferably a bioadhesive patch.

20. The bioadhesive formulation for use according to any one of claims 1 to 19, whereinthe bioadhesive formulation is a mucoadhesive formulation formulated for topical administration onto the mucosal lesion; andthe mucoadhesive formulation prevents microbial growth in the mucosal lesion.

21. The bioadhesive formulation for use according to any one of claims 1 to 20, whereinthe bioadhesive formulation is a dermoadhesive formulation formulated for topical administration onto the dermal lesion; andthe dermoadhesive formulation prevents microbial growth in the dermal lesion.

22. The bioadhesive formulation for use according to any one of claims 1 to 21, for use in prevention or treatment of a microbial infection in the mucosal or dermal lesion caused by microbes selected from the group consisting of bacteria, fungi, viruses and parasites, and any combination thereof.

23. The bioadhesive formulation for use according to any one of claims 1 to 22, for use in prevention or treatment of a microbial infection in the mucosal or dermal lesion, wherein the bioadhesive formulation inhibits microbial growth in the mucosal or dermal lesion, wherein the microbes are selected from the group consisting of bacteria, fungi, viruses and parasites, and any combination thereof.

24. The bioadhesive formulation for use according to claim 23, wherein the bioadhesive formulation inhibits bacterial grow of bacteria, such as Staphylococcus bacteria, preferably Staphylococcus aureus,Streptococcus bacteria, preferably selected from the group consisting of Streptococcus sanguinis, Streptococcus mitis, Streptococcus pyogenes, Streptococcus epidermidis, Streptococcus mutans, Streptococcus salivarius, and any combination thereof, Prevotella bacteria, preferably Prevotella intermedia, Porphyromonas bacteria, preferably Porphyromonas gingivalis, Fusobacterium bacteria, preferably Fusobacterium nucleatum, Treponema bacteria, preferably Treponema denticola, Helicobacter pylori, and / or Pseudomonas aeruginosa, in the mucosal or dermal lesion.

25. The bioadhesive formulation for use according to claim 23 or 24, wherein the bioadhesive formulation inhibits fungal grow, such as Actinomyces fungi, preferably selected from the group consisting of Actinomyces naeslundii, Actinomyces viscosus, and any combination thereof, Candida albicans, Aspergillus fumigatus, Coccidioides immitis, Histoplasma capsulatum, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Geotrichum fungi, Fusarium fungi, Rhodotorula fungi, Saccharomyces fungi and / or Penicillium mameffei, in the mucosal or dermal lesion.

26. The bioadhesive formulation for use according to any one of claim 23 to 25, wherein the bioadhesive formulation inhibits viral grow, such as Herpes simplex viruses 1, (HSV-1), HSV-2, Herpes zoster virus, Epstein-Barr virus (EBV), and / or Cytomegalovirus (CMV), in the mucosal or dermal lesion.

27. The bioadhesive formulation for use according to any one of claim 23 to 26, wherein the bioadhesive formulation inhibits growth of parasites, such as Trichomonas tenax and / or Entamoeba gingivalis, in the mucosal or dermal lesion.

28. The bioadhesive formulation for use according to any one of claims 1 to 27, wherein the bioadhesive formulation is a hygroscopic bioadhesive formulation capable of absorbing fluids corresponding to at least 10 times a weight of the bioadhesive formulation, preferably at least 15 times the weight of the bioadhesive formulation, more preferably 20 times the weight of the bioadhesive formulation, and most preferably 25 times the weight of the bioadhesive formulation.

29. The bioadhesive formulation for use according to any one of claims 1 to 28, wherein the bioadhesive formulation is an absorptive bioadhesive formulation capable of absorbing microbes and / or molecules from the mucosal or dermal lesion.

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