Composition, methods, and uses thereof for acne prevention, reduction, and treatment

Abstract

Compositions and methods are provided for using DispersinB® and a second antimicrobial which does not comprise a bacteriophage or transglutaminase enzyme as a topical anti-acne treatment. The second antimicrobial may be selected from the group consisting of organic peroxide, benzoyl peroxide, tetracyclines such as minocycline and doxycycline, macrolides such as erythromycin and azithromycin, and clindamycin.

Description

COMPOSITION, METHODS, AND USES THEREOF FOR ACNE PREVENTION, REDUCTION, AND TREATMENTCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application is related to and claims any benefit of U.S. Provisional Application Serial No. 63 / 516,385, filed on July 28, 2023, titled "Composition, Methods, and Uses thereof for Acne Prevention, Reduction, and Treatment."TECHNICAL FIELD

[0002] The present invention relates to compositions, methods, and uses thereof for acne prevention, reduction, and treatment.BACKGROUND

[0003] Acne vulgaris (also known as acne) is a chronic inflammatory skin disease that affects 80-85% of the population. Acne usually occurs on the face, chest, shoulders or back. It is most common in teenagers and rarely occurs in people over the age of 50. Although non-life-threatening, acne often causes psychological stress, and may result in permanent physical and mental scarring.

[0004] Acne is caused by dysbiosis of two common bacteria that inhabit the skin, Cutibacterium acnes (Cac) and Staphylococcus epidermidis (Sep). Dysbiosis results in overgrowth of C. acnes, inflammation, and the formation of acne lesions.

[0005] Current acne drugs (antiseptics, antibiotics, beta hydroxy acids, retinoids) exhibit limited effectiveness, and one half to three quarters of acne patients are not satisfied with current over-the-counter (OTC) treatments. Treatment for acne usually lasts 4-12 weeks and requires complex combination treatment regimens that lead to poor adherence which can undermine treatment success.

[0006] The most common topical treatment for active acne lesions is benzoyl peroxide. However, benzoyl peroxide is not always effective and can cause skin irritation. OTC anti-acne washes typically contain 2.5, 4, 5, 6 or 10% benzoyl peroxide. Higher concentrations may be more effective, but can cause more skin irritation, which requires some patients to use lower, less effective doses.

[0007] Skin reactions such as skin irritation, contact dermatitis, pain, irritation, itching, burning, redness, peeling, dryness, scaling, or stinging of the skin can occur during treatment of antimicrobial products recommended for C. acnes infections. Increased and long-term use of antimicrobials can also lead to development of antimicrobial resistance and then make antimicrobial treatments ineffective. As well, conventional acne drugs are not able to prevent new acne lesions.

[0008] DispersinB has been suggested as an adjuvant in the treatment of acne (CA3060415, Compositions Comprising Propionibacterium Acnes Bacteriophages for Treating Acne, see, e.g. paragraphs 0099-0100 and claim 28; CA3181352 Transglutaminase Variants and Applications of Use Thereof, see, e.g. paragraph 31 and claim 30).

[0009] However, DispersinB when tested was found to be ineffective in the treatment of acne (Okuda et al. The composition and structure of biofilms developed by Propionibacterium acnes isolated from cardiacpacemaker devices. Front Microbiol 14 February 2018 Volume 9; Abstract and Figure IB. https: / / doi.org / 10.3389 / fmicb.2018.00182).

[0010] There is therefore a need to provide compositions, methods, and uses thereof that targets dysbiosis of organisms such as Cutibacterium acnes and Staphylococcus epidermidis which are (poly- / V-acetylglucosamine) PNAG-producing organisms, and / or to provide alternatives to antibiotic treatments to reduce the burden of antibiotic resistance and target the very early stages of acne (microcomedogenesis) to prevent new acne lesions.SUMMARY OF THE INVENTION

[0011] It is an embodiment of the present invention to provide compositions, methods, and uses thereof for acne prevention, reduction, and treatment.

[0012] In one embodiment, the present invention relates to compositions for reducing active acne lesions and preventing new acne lesions. In aspects, the compositions are formulated for topical administration.

[0013] In one embodiment, the composition comprises one or more enzymes for hydrolyzing linear polymers of / V-acetyl-D-glucosamines to prevent biofilm formation by bacteria. In aspects, the bacteria are C. acnes and S. epidermidis.

[0014] In one embodiment, there is provided a use of a composition comprising one or more enzymes for hydrolyzing linear polymers of N- acetyl-D-glucosamines as an anti-acne treatment. In one aspect, the one or more enzymes for hydrolyzing linear polymers of / V-acetyl-D-glucosamines comprises DispersinB®.

[0015] In one embodiment, there is provided a method of treating acne comprising applying a composition comprising Dispersing®; and applying an organic peroxide-containing wash after the application of the Dispersing®. In aspects, the composition is formulated in a form of liquid, film, tape, or gel.BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Fig ure 1 is photo showing inhibition of C. acnes biofilm formation by Dispersing® 80 pg / ml in accordance with an embodiment of the invention;

[0017] Fig ure 2a is a graph showing growth of S. epidermidis, either alone or in co-culture with C. acnes, in aerobic and anaerobic conditions over time;

[0018] Fig ure 2b is a graph showing growth of C. acnes, either alone or in co-culture with S. epidermidis, in aerobic and anaerobic conditions over time;

[0019] Fig ure 3a is a graph showing the survival of S. epidermidis when co-cultured with C. acnes in an aerobic biofilm and then treated with 80 pg / ml Dispersing®, 1% benzoyl peroxide (gP), or phosphate buffered saline (PgS), followed by a second treatment with the same agents in the indicated combinations;

[0020] Fig ure 3b is a graph showing the survival of C. acnes when cocultured with S. epidermidis in an aerobic biofilm and then treated with 80 pg / ml Dispersing®, 1% benzoyl peroxide (gP), or phosphate buffered saline (PgS), followed by a second treatment with the same agents in the indicated combinations;

[0021] Fig ure 4a is a graph showing the survival of C. acnes alone (without S. epidermidis) in the presence of benzoyl peroxide (70 pg / ml) (panel A);

[0022] Fig ure 4b is a graph showing the survival of C. acnes alone (without S. epidermidis) in the presence of benzoyl peroxide (140 pg / ml);

[0023] Fig ure 5a is a graph showing the survival of S. epidermidis when co-cultured with C. acnes in an aerobic biofilm and then treated with 80 pg / ml DispersinB®, 2.5% benzoyl peroxide (BP2.5), 1% benzoyl peroxide (BP1), or phosphate buffered saline (PBS), followed by a second treatment with 1% benzoyl peroxide or phosphate buffered saline in the indicated combinations;

[0024] Fig ure 5b is a graph showing the survival of C. acnes when cocultured with S. epidermidis in an aerobic biofilm and then treated with 80 pg / ml DispersinB®, 2.5% benzoyl peroxide (BP2.5), 1% benzoyl peroxide (BP1), or phosphate buffered saline (PBS), followed by a second treatment with 1% benzoyl peroxide or phosphate buffered saline in the indicated combinations;

[0025] Fig ure 6a is a graph showing the survival of S. epidermidis when co-cultured with C. acnes in an aerobic biofilm and then treated with 80 pg / ml DispersinB®, 40 pg / ml DispersinB®, or phosphate buffered saline (PBS), followed by a second treatment with 2.5% benzoyl peroxide (BP2.5), 1% benzoyl peroxide (BP1), or phosphate buffered saline in the indicated combinations;

[0026] Fig ure 6b is a graph showing the survival of C. acnes when co- cultured with S. epidermidis in an aerobic biofilm and then treated with 80 pg / ml DispersinB®, 40 pg / ml DispersinB®, or phosphate buffered saline (PBS), followed by a second treatment with 2.5% benzoyl peroxide (BP2.5),1% benzoyl peroxide (BP1), or phosphate buffered saline in the indicated combinations;

[0027] Fig ure 7a is a graph showing the survival of S. epidermidis when co-cultured with C. acnes in an aerobic biofilm and then treated with 20 pg / ml DispersinB®, 10 pg / ml DispersinB®, 5 pg / ml DispersinB®, or phosphate buffered saline (PBS), followed by a second treatment with 0.5% benzoyl peroxide (B.5), 0.1% benzoyl peroxide (B. l), or phosphate buffered saline in the indicated combinations;

[0028] Fig ure 7b is a graph showing the survival of C. acnes when cocultured with S. epidermidis in an aerobic biofilm and then treated with 20 pg / ml DispersinB®, 10 pg / ml DispersinB®, 5 pg / ml DispersinB®, or phosphate buffered saline (PBS), followed by a second treatment with 0.5% benzoyl peroxide (BP.5), 0.1% benzoyl peroxide (BP. l), or phosphate buffered saline in the indicated combinations;

[0029] Fig ure 8a is a graph showing the survival of S. epidermidis when co-cultured with C. acnes in an aerobic biofilm and then treated with 80 pg / ml DispersinB® or phosphate buffered saline (PBS) for 15 min, followed by a second treatment with 1% benzoyl peroxide (BP1) or phosphate buffered saline for 2, 7, or 12 min;

[0030] Fig ure 8b is a graph showing the survival of C. acnes when co- cultured with S. epidermidis in an aerobic biofilm and then treated with 80 pg / ml DispersinB® or phosphate buffered saline (PBS) for 15 min, followed by a second treatment with 1% benzoyl peroxide (BP1) or phosphate buffered saline for 2, 7, or 12 min;

[0031] Fig ure 9 is a graph showing growth of C. acnes in the presence of 20 pg / ml DispersinB® and / or 20 pg / ml tetracycline; and

[0032] Figure 10 is a flow chart of a method for acne prevention, reduction, and treatment.

[0033] Fig ure 11 is a collection of graphs showing six different staphylococcal strains each enables four different Cutibacterium sp. strains to grow in aerobic conditions.DETAILED DESCRIPTION

[0034] Reference will be made below in detail to exemplary embodiments of the invention, examples of which are illustrated in the accompanying figures.

[0035] "benzoyl peroxide" is an example organic peroxide antiseptic that exhibits antimicrobial activity against Cutibacterium acnes, a causative agent acne vulgaris.

[0036] "DispersinB" is an example enzyme that degrades poly- / V- acetylglucosamine (PNAG).

[0037] "DispersinB® Human Wound Gel" refers to a hydrogel formulation comprising DispersinB® and Pluronic F-127 surfactant.

[0038] "PNAG" refers to an extracellular polysaccharide that mediates biofilm formation by numerous bacterial pathogens including Cutibacterium acnes and Staphylococcus epidermidis, two bacteria involved in the pathogenesis of acne vulgaris.ExamplesExample 1 - DispersinB® inhibits C. acnes biofilm formation.

[0039] As shown in Figure 1, C. acnes was cultured aerobically in glass tubes in 1 ml of Tryptic Soy broth supplemented with 0 or 80 pg / ml of DispersinB®. Tubes were photographed after 3 d. Triplicate tubes for each treatment are shown. DispersinB® inhibits C. acnes biofilm formation as evidenced by the absence of biofilm on the sides of the tubes containing DispersinB®. DispersinB® also inhibits biofilm formation by C. acnes in polypropylene tubes.Example 2 - S. epidermidis promotes C. acnes growth under aerobic conditions when cultured in a dual-species biofilm.

[0040] Fig ures 2a and 2b shows growth of S. epidermidis and C. acnes over time. S. epidermidis (Sep) and C. acnes (Cac) were cultured alone (blue and green) or together (1 : 1 ratio; red and violet) under anaerobic (green and violet) or aerobic (blue and red) conditions. CFU / tube values for each species were enumerated after 72 h. Values show mean CFU / tube for duplicate tubes. C. acnes is an aerotolerant anaerobe that only grows under anaerobic conditions but is not killed by oxygen; however as shown in Figures 2a and 2b, co-cultured with S. epidermidis, C. acnes will grow under aerobic conditions.

[0041] Without being limited to any particular theory, it is believed that S. epidermidis biofilm creates an anaerobic environment where C. acnes can grow. C. acnes growth under aerobic conditions has not previously been observed. Additionally, aerobic S. epidermidis / C. acnes dual-species biofilms would appear to mimic the relationship of these species in acneic hair follicles and thus provides a useful screen for novel anti-acne agents.

[0042] The aerobic S. epidermidis / C. acnes dual-species biofilm model was used in the subsequent experiments shown in Figures. 3-7.Example 3 - Pre-treatment of S. epidermidis / C. acnes dual-species and C. acnes mono-species biofilms with DispersinB® sensitizes bacteria killing by benzoyl peroxide.

[0043] To determine whether DispersinB® sensitizes PNAG-producing bacteria to killing by antiseptics, disinfectants and antibiotics, S. epidermidis / C. acnes dual-species biofilms were treated with phosphate buffered saline (PBS) or 80 pg / ml DispersinB® in PBS for 15 min, then rinsed and treated with 1% benzoyl peroxide for 15 min. The surviving biofilm cells were counted for each species individually. It should be noted that a 1% benzoyl peroxide is below the 2.5% lowest effective dose used in OTC anti-acne products.

[0044] 24-h-old aerobic S. epidermidis / C. acnes dual-species biofilms were rinsed and treated with phosphate buffered saline (PBS), 80 pg / ml DispersinB (DSP), or 1% benzoyl peroxide (BP). After 15 min, biofilms were rinsed and subjected to a second 15-min treatment with PBS, DSP or BP in the indicated combinations.

[0045] As shown in Figure 3, graphs show CFU / tube values for each species from triplicate tubes for each treatment. DispersinB® treatment (DSP) (purified DispersinB® enzyme) alone caused a 1-log reduction in S. epidermidis CFUs, and a 2-log reduction in C. acnes CFUs, whereas DispersinB® + benzoyl peroxide caused a 2-log reduction in S. epidermidis CFUs, and a 5-log reduction in C. acnes CFUs. Benzoyl peroxide treatment alone caused no reduction in S. epidermidis CFUs and a 1-log reduction in C. acnes CFUs.

[0046] From these experiments, it is clear that DispersinB® sensitizes C. acnes biofilms to killing by benzoyl peroxide.

[0047] In contrast, pre-treatment of S. epidermidis / C. acnes dualspecies biofilms with 1% benzoyl peroxide followed by DispersinB® treatment inhibited the ability of DispersinB® to detach S. epidermidis and did not sensitize C. acnes to detachment by DispersinB®.

[0048] C. acnes alone (without S. epidermidis) was cultured anaerobically in the presence of 20 pg / ml DispersinB® and / or 70 (panel A) or 140 (panel B) pg / ml benzoyl peroxide. CFUs were measured after 48 h. As shown in Figure 4, similarly, DispersinB® also sensitized C. acnes monospecies to killing by low doses of benzoyl peroxide.

[0049] From these experiments, it is clear that DispersinB® sensitizes S. epidermidis / C. acnes dual-species and C. acnes mono-species biofilms to killing by benzoyl peroxide.Example 4 - DispersinB® + 1% benzoyl peroxide kills C. acnes biofilms as efficiently as 2.5% benzoyl peroxide alone.

[0050] An experiment to compare the effectiveness of DispersinB® + 1% benzoyl peroxide to that of 1% and 2.5% benzoyl peroxide alone at eradicating S. epidermidis / C. acnes dual-species biofilms is shown in Figure 5. 24-h-old aerobic S. epidermidis / C. acnes dual-species biofilms were rinsed and treated with phosphate buffered saline (PBS), 80 pg / ml DispersinB® (DSP), or 1% or 2.5% benzoyl peroxide (BP1 and BP2.5, respectively). After 15 min, biofilms were rinsed and subjected to a second 15-min treatment with PBS or BP1 in the indicated combinations. Graphs show CFU / tube values for each species from triplicate tubes for each treatment.

[0051] For both S. epidermidis and C. acnes, the DispersinB® + 1% benzoyl peroxide combination treatment killed significantly more bacteria than either 1% or 2.5% benzoyl peroxide treatment alone.Example 5 - DispersinB® Human Wound Gel sensitizes C. acnes biofilms to killing by benzoyl peroxide.

[0052] In the experiment as shown in Figure 6, 24-h-old aerobic S. epidermidis / C. acnes dual-species biofilms were rinsed and treated with phosphate buffered saline (PBS), DispersinB® Human Wound Gel containing 40 pg / ml DispersinB® (D40), or DispersinB® Human Wound Gel containing 80 pg / ml DispersinB® (D80). After 15 min, biofilms were rinsed and subjected to a second 15-min treatment with PBS, 1% benzoyl peroxide (BP1), or 2.5% benzoyl peroxide (BP2.5) in the combinations shown. Graphs show CFU / tube values for each species from duplicate tubes for each treatment. S. epidermidis / C. acnes dual-species biofilms were treated with DispersinB® Human Wound Gel (40 or 80 pg / ml) followed by benzoyl peroxide (1% or 2.5% concentrations). In these assays, treatment times was a 15-min enzyme treatment step followed by a 15-min benzoyl peroxide treatment step or PBS step.

[0053] As shown, DispersinB® Human Wound Gel treatment alone resulted in a 1-log reduction on S. epidermidis cells, and a 2-log reduction in C. acnes cells.

[0054] Benzoyl peroxide treatment alone resulted in no significant decrease in S. epidermidis CFUs, and a 1-log reduction in C. acnes CFUs. However, pre-treatment of biofilms with DispersinB® Human Wound Gel (40 or 80 pg / ml) followed by benzoyl peroxide (1.0 or 2.5%) resulted in reduction of C. acnes CFUs to below detectable levels (< 50 CFU / tube).Example 6 - DispersinB® Human Wound Gel sensitizes C. acnes biofilms to benzoyl peroxide killing.

[0055] To determine the lowest doses of DispersinB® and benzoyl peroxide that effectively eradicate C. acnes from S. epidermidis / C. acnesdual-species biofilms, biofilms were treated with DispersinB® Human Wound Gel containing 20, 10 or 5 pg / ml DispersinB® followed by benzoyl peroxide at 0.5 or 0.1%.

[0056] 24-h-old aerobic S. epidermidis / C. acnes dual-species biofilms were rinsed and treated with PBS or DispersinB® Human Wound Gel containing 20, 10 or 5 pg / ml DispersinB® (D20, D10, and D5 respectively). After 15 min, biofilms were rinsed and treated with PBS or benzoyl peroxide at 0.5 or 0.1% (BP.5 and BP. l, respectively). Graphs show CFU / tube values for each species from duplicate tubes for each treatment. As shown in Figure 7, DispersinB® Human Wound Gel treatment alone (20, 10 or 5 pg / ml) resulted in a 1-log reduction on S. epidermidis cells, and a 2-log reduction in C. acnes cells.

[0057] Benzoyl peroxide treatment alone resulted in no significant decrease in S. epidermidis CFUs, and 2-log and 1-log reductions in C. acnes CFUs for 0.5% and 0.1%, respectively.

[0058] However, pre-treatment of biofilms with DispersinB® Human Wound Gel (20, 10 or 5 pg / ml) followed by benzoyl peroxide at 0.5% resulted in reduction of C. acnes CFUs to below detectable levels (< 50 CFU / tube).Example 7 - Time course of benzoyl peroxide killing

[0059] To determine whether a shorter benzoyl peroxide contact time can eradicate DispersinB® Human Wound Gel-treated aerobic S. epidermidis / C. acnes dual-species biofilms, biofilms were rinsed and treated with PBS or DispersinB® Human Wound Gel containing 80 pg / ml DispersinB® for 15 min, and then rinsed and treated with PBS or 1% benzoyl peroxide for 2, 7, or 12 min.

[0060] 24-h-old aerobic S. epidermidis / C. acnes dual-species biofilms were rinsed and treated with PBS or DispersinB® Human Wound Gel containing 80 pg / ml DispersinB® (DWG). After 15 min, biofilms were rinsed and subjected to a second treatment with PBS or 1% benzoyl peroxide (BP1) for 2, 7, or 12 min. Graphs show log CFU / tube values for each species from a single tube for each treatment. As shown in Figure 8, a significant reduction in C. acnes CFUs occurred after 2 min in biofilms treated with DispersinB® Human Wound Gel + benzoyl peroxide.Example 8 - DispersinB® sensitizes C. acnes to killing by tetracycline

[0061] An experiment to measure the ability of DispersinB® to sensitize C. acnes alone (without S. epidermidis') to killing by tetracycline is shown in Figure 9. Growth of C. acnes HL043PA1 in the presence of 0 or 20 pg / ml DispersinB®, and / or 0.2 pg / ml tetracycline was measured.

[0062] As shown, tetracycline alone was ineffective in inhibiting C. acne. However, there was a significant reduction in C. acnes CFUs when treatment of DispersinB® was used prior to treatment with tetracycline.Example 9 - DispersinB® sensitizes C. acnes to killing by tetracyclineThe same dual-species biofilm model that was used in the previous experiments was used to investigate multiple different staphylococcal strains ability to enable different Cutibacterium sp. strains to grow in air.Aerobic growth o Cutibacterium spp. in 15-mL conical-bottom polypropylene tubes. Four Cutibacterium spp. (species and strain names indicated at the right) were cultured alone anaerobically (solid black line), alone aerobically (dashed black line), or co-cultured aerobically with each of the staphylococcal species and strain indicated at the top (red lines). Graphs show mean C. acnes CFU / tube values after 40 h for triplicate tubes. Errorbars (ca. 10% sd) were omitted for clarity. All assays were performed on three separate occasions with nearly identical results. Data from representative experiments are shown.Figure 11 shows growth of four different Cutibacterium spp. strains in 15-mL conical-bottom polypropylene tubes. In this experiment, four Cutibacterium spp. strains (species and strain names indicated at the right) were cultured alone anaerobically (solid black line), alone aerobically (dashed black line), or co-cultured aerobically with the staphylococcal species and strain indicated at the top (red lines). Graphs show mean C. acnes CFU / tube values after 40 h for triplicate tubes. Error bars (ca. 10% sd) were omitted for clarity.Method and uses for acne prevention, reduction, and treatment

[0063] Thus a two-step topical skincare regimen is useful for both the reduction of active acne lesions and the prevention of new acne lesions. The first step is application of Dispersing to the skin, and the second step is application of an antimicrobial to the skin. There is a 10-15 min waiting period between applications to allow sufficient contact time for the enzyme to work.

[0064] As demonstrated in the above Examples, pre-treatment of skin with Dispersing® Human Wound Gel sensitizes C. acnes to benzoyl peroxide treatment, thereby increasing the effectiveness of benzoyl peroxide treatments against active acne lesions. Accordingly, in some aspects, the minimum effective dose of benzoyl peroxide can be reduced from 2.5% to 0.5% and this consequently reduces skin irritation or the chance of skin irritation.

[0065] The ability of Dispersing® to inhibit C. acnes biofilm formation will prevent the formation of early acne lesions which progress to active acnelesions. DispersinB® can be formulated into a variety of different physical formulation including, liquids, films, tapes, or gels.

[0066] The ability of DispersinB® or DispersinB® Human Wound Gel to inhibit S. epidermidis biofilm formation will prevent early acne lesions and reduce active acne lesions by preventing S. epidermidis biofilm formation and its ability to promote C. acnes growth. Compositions comprising DispersinB® will be useful in pretreatment methods in an aqueous, emulsion based wash, leave on, or cosmetics to maintain skin homeostasis and prevent acne.

[0067] Fig ure 10 is an example anti-acne method of treatment 100. Each block represents one or more steps and the order is for illustration purposes only. Additional blocks may be added or fewer blocks may be utilized without departing from the disclosure. At block 102, once daily in the evening, wash face with skin cleanser, such as Cetaphil. At block 104, apply DispersinB® Human Wound Gel to one or more areas. At block 106, wait about 2 to about 15 min. At block 108, wash face with benzoyl peroxide anti-acne wash.

[0068] Since both anionic and non-ionic detergents detach dual-species biofilms, a third step consisting of a detergent may increase the effectiveness of the procedure. A three-step procedure can be carried out in different orders, such as detergent / DispersinB® Human Wound Gel / benzoyl peroxide, or DispersinB® Human Wound Gel / detergent / benzoyl peroxide.

[0069] Antibiofilm agents are a new and preferred approach to treating acne, and DispersinB® is a unique antibiofilm agent because it specifically targets PNAG-producing organisms. Second, alternatives to antibiotic treatments are preferable in order to reduce the burden of antibiotic resistance. DispersinB® does not promote resistance because it does not kill bacteria or inhibit their growth. Resistance to benzoyl peroxide has notbeen observed. Third, Dispersing® is novel therapeutic because it targets the very early stages of acne (microcomedogenesis), and may therefore be more effective at preventing new acne lesions than conventional acne drugs. An antibiofilm approach to treating acne has the potential to improve clinical outcomes and may provide an exciting and novel treatment option for acne patients.Method and uses to prevent medical device infections

[0070] C. acnes forms biofilms on implanted medical devices such as prosthetic joints, cardiac pacemakers, and cerebrospinal fluid shunts where the source of bacteria in these infections is the patient's own skin.

[0071] In an embodiment there is provided a method for pre-surgical skin antisepsis for subjects receiving implanted medical devices comprising applying a Dispersing® or Dispersing® composition before the step of surgical implantation of the medical devices.Method and uses to maintain skin microbiome equilibrium and to prevent skin infections and / or acne

[0072] C. acnes and S. epidermidis are part of the normal skin flora and have a symbiotic relationship.

[0073] In an embodiment there is provided a method comprising applying a Dispersing® or Dispersing® composition in combination with cleansers, cosmetics and medicants for acne and atopic dermatitis.

[0074] The embodiments of the present application described above are intended to be examples only. Those of skill in the art may effect alterations, modifications and variations to the particular embodiments without departing from the intended scope of the present application. Inparticular, features from one or more of the above-described embodiments may be selected to create alternate embodiments comprised of a subcombination of features which may not be explicitly described above. In addition, features from one or more of the above-described embodiments may be selected and combined to create alternate embodiments comprised of a combination of features which may not be explicitly described above. Features suitable for such combinations and subcombinations would be readily apparent to persons skilled in the art upon review of the present application as a whole. Any dimensions provided in the drawings are provided for illustrative purposes only and are not intended to be limiting on the scope of the invention. The subject matter described herein and in the recited claims intends to cover and embrace all suitable changes in technology.

Claims

WE CLAIM :

1. Use of a composition comprising a Dispersing® and a second antimicrobial which does not comprise a bacteriophage or transglutaminase enzyme as a topical anti-acne treatment.

2. Use of a composition comprising a DispersinB®-containing formulation as a topical anti-acne treatment further comprising administration of a second antimicrobial effective against acne which does not comprise a bacteriophage or transglutaminase enzyme.

3. Use according to claim 1 or 2 wherein the second antimicrobial is selected from the group consisting of organic peroxide, benzoyl peroxide, tetracyclines such as minocycline and doxycycline, macrolides such as erythromycin and azithromycin, and clindamycin.

4. Use according to claims 2 or 3 wherein the DispersinB®-containing formulation is administered prior to administration of the second antimicrobial.

5. Use according to claim 4 wherein the DispersinB®-containing formulation is administered 2-15 minutes prior to administration of the second antimicrobial.

6. Use according to any of claims 1 to 5 for preventing, reducing, or removing acne lesions where the lesions are one or more of microcomedos, open comedos (blackheads), closed comedos (whiteheads) and pimples.

7. Use according to any of claims 1 to 5 for preventing, reducing, or removing bacterial biofilms and sebaceous plugs that form inside the hair follicle.

8. Use according to any of claims 1 to 7 for preventing, reducing, or removing colonization of the hair follicle by Cutibacterium acnes and Staphylococcus bacteria.

9. Use according to claim 8 wherein the Cutibacterium acnes is selected from the group consisting of strain 2, strain 7, strain 14, and strain 42.

10. Use according to claim 8 or 9 wherein the Staphylococcus is selected from the group consisting of S. epidermidis, S. hominus, and S. capitus.

11. Use according to claim 10 wherein the S. epidermidis is selected from the group consisting of strain JK4, strain JK6, and strain JK7.

12. A composition for acne prevention, reduction, and treatment comprising DispersinB® and a second antimicrobial, wherein said second antimicrobial is not a bacteriophage or a transglutaminase enzyme.

13. The composition of claim 12 wherein DispersinB®-containing formulation is DispersinB® Human Wound Gel formulation.

14. The composition of claim 12 or 13 wherein the DispersinB®-containing formulation comprises a concentration of DispersinB® enzyme from about 5 to about 80 pg / ml.

15. The composition of any one of claims 12 to 14 further comprising a benzoyl peroxide where the concentration of benzoyl peroxide ranges from about 0.5 - about 10%; about 0.25 - about 10%, about 0.5 - about 5%, or about 0.5 - 2.5%.

16. A method for treating acne comprising applying a DispersinB® formulation to skin; and applying a benzoyl peroxide-containing wash to the skin after the application of the DispersinB formulation.

17. The method according to claim 13 where the DispersinB formulation is left on the skin for about 2 to about 15 min prior to the application of the benzoyl peroxide-containing wash.

18. The method according to claim 16 or 17 where the benzoyl peroxide- containing wash is left on the skin for about 2 to about 15 min.

19. A method for treating C. acnes- or S. epiderm idis- associated skin infections comprising applying a DispersinB® formulation and applying a reduced dose and / or shorter duration of a second antimicrobial, said reduced dose and / or shorter duration being less than required to be effective when using said second antimicrobial alone.

20. The method of claim 19 wherein the application of the DispersinB® composition precedes the application of the second antimicrobial.

21. The method of claim 19 wherein the application of the DispersinB® composition is followed by applying a surfactant before the application of the second antimicrobial.

22. A method for treating C. acnes infections comprising applying one or more of a surfactant, buffering agents, and preservatives, to a skin and applying a DispersinB® or DispersinB® composition to the skin and subsequently applying a second antimicrobial to the skin.

23. The method of any one of claims 19 to 22 wherein the second antimicrobial is is selected from the group consisting of organic peroxide, benzoyl peroxide, tetracyclines such as minocycline anddoxycycline, macrolides such as erythromycin and azithromycin, and clindamycin.

24. The method of claim 22 or 23 wherein an amount of said second antimicrobial is about 20 times less than an amount of said second antimicrobial needed without prior application of DispersinB® or DispersinB® composition.

25. The method of claim 22 or 23 wherein the duration of the application of the second antimicrobial is for a shorter time than a duration that the second antimicrobial is needed to be left on the skin without prior application of the DispersinB® or DispersinB® composition26. The method of claim 25 wherein the shorter time is up to about 4 times shorter.

27. The method of any one of claims 22 to 26 wherein the surfactant is an anionic or a non-ionic detergent.

28. The method of any one of claims 22 to 26 wherein the C. acnes- or S. epidermidis-asso ated skin infections is selected from one or more of blepharitis, endophthalmitis, corneal ulcers, sarcoidosis, and atopic dermatitis29. A method for treating C. acnes infections comprising applying a DispersinB® or DispersinB® composition and subsequently applying one or more of a surfactant, a buffering agent, and a preservative, and subsequently applying an organic peroxide.

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