Compositions comprising propionibacterium acnes bacteriophages for treating acne

Abstract

To provide a composition for preventing or treating acne.SOLUTION: Provided is a composition comprising at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound, and a pharmaceutically acceptable carrier.SELECTED DRAWING: Figure 5

Description

[Technical field]

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 488,326, filed April 21, 2017, which is hereby incorporated by reference in its entirety for all purposes.

[0002] This invention was made with Government support under Grant No. 1R43AR068172-01 awarded by the National Institutes of Health. The Government has certain rights in this invention.

[0003] The contents of the text file named "052004-503001WO_SEQUENCE LIST.TXT", which was created on April 20, 2018, is 101,782 bytes in size, and is hereby incorporated by reference in its entirety. [Background technology]

[0004] Acne is a near-universal condition that affects more than 80% of all people worldwide. This chronic skin condition is complex, but the primary pathogen is Propionibacterium acnes, whose overgrowth leads to the inflammation that causes comedones. Despite a clear need for innovation, there have been no new comedonal medications for over 30 years. Current treatments, including benzoyl peroxide and antibiotics, are highly ineffective, and the most effective treatment, isotretinoin, is limited to a small proportion of patients due to dangerous side effects (including birth defects, liver damage and suicide).

[0005] New methods and compositions for treating acne are needed. Summary of the Invention [Problem to be solved by the invention]

[0006] In particular, provided herein are compositions, combinations, systems and methods for preventing or treating acne.

[0007] In aspects, provided herein are compositions comprising, consisting essentially of, or consisting of at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound, and a pharma- ceutically acceptable carrier.

[0008] In an aspect, provided herein is a composition comprising at least one Propionibacterium acnes bacteriophage, no more than one anti-acne compound, and a pharma- ceutically acceptable carrier.

[0009] In an aspect, provided herein is a composition comprising an active ingredient consisting of at least one Propionibacterium acnes bacteriophage and no more than one anti-acne compound, and a pharma- ceutically acceptable carrier.

[0010] In an aspect, provided herein is a composition comprising at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound, and a pharma- ceutically acceptable carrier, the composition being free of probiotic bacteria.

[0011] In an aspect, provided herein is a composition comprising a Propionibacterium acnes bacteriophage and an enzyme.

[0012] In aspects, provided herein are combinations comprising, consisting essentially of, or consisting of at least one Propionibacterium acnes bacteriophage and at least one anti-acne compound, wherein each of the at least one Propionibacterium acnes bacteriophage and the at least one anti-acne compound is in a composition that further comprises a pharma- ceutically acceptable carrier.

[0013] In an aspect, provided herein is a combination comprising a Propionibacterium acnes bacteriophage and an enzyme.

[0014] In an aspect, provided herein is a method of preventing or treating acne in a subject in need thereof, comprising administering an effective amount of a composition or combination provided herein. [Brief description of the drawings]

[0015] [Figure 1] P. acnes (acne-causing, left half plate) or P. granulosum (symbiotic, right half plate) bacteria were seeded onto RCM agar Petri dishes. A sterile half-pad soaked with either minocycline or PHIT-101 (107 pfu / mL) was placed onto each plate. After 3 days of anaerobic incubation at 37°C, a killer zone (arrow) appeared, indicating that minocycline kills both pathogenic and commensal bacteria, while PHIT-101 kills acne-causing bacteria and spares commensal P. granulosum. [Diagram 2] A synthetic skin microbiome containing P. acnes, P. granulosum, and P. avidum was grown to confluence in a test tube. The synthetic skin microbiome was then incubated for 48 h in the presence or absence of PHIT-101. The composition ratio of the three bacterial species was quantified by NGS sequencing of 16S amplicons of washed bacterial pellets using an Illumina MiSeq platform. PHIT-101 was able to almost completely eradicate acne-causing P. acnes while not affecting the growth of the other two commensal species. [Diagram 3] Biofilm production in P. acnes is highly variable. Ninety-six strains of P. acnes were grown in 96-well polystyrene microtiter plates to stimulate biofilm production, and the biofilm produced by each strain was quantified. The variability exhibited within this set of strains demonstrates the need to quantify biofilm formation under growth conditions that are more similar to those found in human hair pores. [Figure 4]Screening to select enzymes capable of degrading P. acnes biofilm. P. acnes was grown in polystyrene microtiter plates to stimulate biofilm production. Enzymes were added to the wells at 0.01 mg / mL and incubated at 30° C. for 30 min. The degraded biofilm was washed away with phosphate-buffered saline (PBS) and the remaining biofilm in each well was quantified by staining with crystal violet and recording the absorbance at 590 nm. Proteases such as proteinase K and subtilisin showed good activity, and dispersin was the best glycoside depolymerase tested. [Diagram 5] Augmentation of phage with biofilm-degrading enzymes (BDEs) significantly increases bactericidal activity. Sessile P. acnes cells were incubated with PBS (untreated), PHIT-101, or PHIT-101 plus dispersin. Cell viability was measured using CellTiter-Blue reagent and fluorescence was recorded at 560Ex / 590Em. PHIT-101 was not as effective at killing P. acnes as in liquid cultures, but the addition of the biofilm-degrading enzyme dispersin significantly increased bactericidal activity to levels similar to those in liquid cultures. [Figure 6] Probiotic strains produce low levels of lipase in attached culture. Probiotic P. acnes strains with known genotypes were grown under biofilm conditions in microtiter plates. After 72 h of growth, culture supernatants were filter sterilized and incubated with 4-MU palmitate for 4 h at 37°C to determine extracellular lipase production. Lipase production by the probiotic strain (Pr#X) was very low compared to the pathogenic strain, indicating a relatively low proinflammatory potential. [Figure 7]Probiotic strains exhibit significantly less adhesion to epithelial cells compared to pathogenic P. acnes. Selected probiotic strains were incubated with confluent A-431 epithelial cells (MOI 10). After washing the wells, cells were lifted using 0.1% Tween 80 and plated on BHI plates. After 72 hours of anaerobic incubation, colonies were counted. Data revealed that probiotic strains exhibited significantly less binding to epithelial cells (*p<0.05, **p<0.005). [Figure 8A] Lower proinflammatory potential of probiotic strains in a mouse ear inflammation model. CBA / J mice (5 mice per cohort) were injected with P. acnes strains and cytokine analysis was performed on day 5. Probiotic strain Pr#C showed significantly lower levels (*p<0.05, **p<0.01, ***p<0.0001) of the proinflammatory cytokines IL-1β (Figure 8A), IL-6 (Figure 8B), IL-17 (Figure 8C) and TNFα (Figure 8D) compared to the pathogenic strains. Pr-C has the ProII 16S sequence. [Figure 8B] Lower proinflammatory potential of probiotic strains in a mouse ear inflammation model. CBA / J mice (5 mice per cohort) were injected with P. acnes strains and cytokine analysis was performed on day 5. Probiotic strain Pr#C showed significantly lower levels (*p<0.05, **p<0.01, ***p<0.0001) of the proinflammatory cytokines IL-1β (Figure 8A), IL-6 (Figure 8B), IL-17 (Figure 8C) and TNFα (Figure 8D) compared to the pathogenic strains. Pr-C has the ProII 16S sequence. [Figure 8C] Lower proinflammatory potential of probiotic strains in a mouse ear inflammation model. CBA / J mice (5 mice per cohort) were injected with P. acnes strains and cytokine analysis was performed on day 5. Probiotic strain Pr#C showed significantly lower levels (*p<0.05, **p<0.01, ***p<0.0001) of the proinflammatory cytokines IL-1β (Figure 8A), IL-6 (Figure 8B), IL-17 (Figure 8C) and TNFα (Figure 8D) compared to the pathogenic strains. Pr-C has the ProII 16S sequence. [Figure 8D] Lower proinflammatory potential of probiotic strains in a mouse ear inflammation model. CBA / J mice (5 mice per cohort) were injected with P. acnes strains and cytokine analysis was performed on day 5. Probiotic strain Pr#C showed significantly lower levels (*p<0.05, **p<0.01, ***p<0.0001) of the proinflammatory cytokines IL-1β (Figure 8A), IL-6 (Figure 8B), IL-17 (Figure 8C) and TNFα (Figure 8D) compared to the pathogenic strains. Pr-C has the ProII 16S sequence. [Figure 9] P. acnes strains have different lipase profiles in planktonic and sessile cultures. A set of two pathogenic (Path-1, Path-2) and two probiotic (Pr-1 to Pr-6) P. acnes strains were evaluated for lipase production in planktonic (grey bars) and sessile (black bars) cultures. Lipase production of the probiotic strains was not significantly different from the pathogenic strains in liquid (planktonic) cultures, but lipase yields in attached cultures were consistently lower compared to the corresponding pathogenic cultures. Interestingly, variability in lipase production between the probiotic strains was observed. The strain with the lowest lipase activity was selected. [Figure 10] illustrates the life cycle of an exemplary bacteriophage. Counterclockwise from bottom left: The phage particle recognizes and adsorbs to the surface of a host bacterium. The phage genome is injected into the bacterium. In the lysogenic cycle, this DNA integrates into the bacterial genome and replicates along with it for several cycles. In the lytic cycle, the phage genome does not integrate, but continues to hijack the host machinery to replicate the phage genome and structural components of the phage. The fully assembled phage then lyses the cell, typically by producing endolysins and holins during the later stages of infection. At this point, the released phage is free to seek out and infect a new host bacterium, initiating the lytic cycle once more. [Figure 11]illustrates the formation of an exemplary bacterial biofilm. Bacterial cells land and adhere to a surface with favorable conditions for growth. They replicate and form colonies, eventually triggering biofilm formation at a certain threshold cell density (quorum). Biofilms contain a mixture of polysaccharides, proteins, DNA and lipids in varying proportions. Biofilms are physical barriers that protect bacterial colonies from harsh external conditions and confer resistance to antibiotics, toxins and immune cells. [Figure 12] illustrates an embodiment of the action of the three components in unison; their effects are depicted sequentially for illustration. Inflamed comedones are typically clogged with biofilms (A) produced by overgrown P. acnes along with commensal skin bacteria (B). Biofilm-degrading enzymes (Lightning Bolt) disrupt the P. acnes biofilm, allowing better access to the other components. Next, bacteriophages (Hexagon) edit or specifically kill pathogenic P. acnes, clearing the infection. Finally, probiotic bacteria (C) colonize the pores and occupy the pathogen niche, preventing the pathogen from growing back, and rebalancing the microbiome to a healthy state. [Figure 13] FIG. 1 is a schematic diagram of a non-limiting probiotic bacteria screening process. [Figure 14] 1 is a graph showing that the pathogenic strains caused significantly more ear inflammation than the PBS control, while the lead probiotic strain Pr-C induced ear inflammation that was not significantly different from the PBS control. [Figure 15] 1 is a graph showing that phage remain stable in the presence of low (0.5% w / v) and high (2% w / v) concentrations of salicylic acid. [Figure 16] is a graph showing that phage lose viability in the presence of benzoyl peroxide over a 60 day period. The rate of decrease in phage viability is greater at the higher concentration (10% w / v) compared to the lower concentration (2.5% w / v). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] In particular, provided herein are compositions, combinations, methods and systems for treating and preventing acne.

[0017] Salicylic acid and benzoyl peroxide are the most commonly used anti-acne agents in over-the-counter (OTC) products. The stability of phages in combination with these anti-acne agents is unknown, especially since phages are widely divergent in their stability and response to external physical and chemical factors. The redox properties of benzoyl peroxide and sulfur could potentially cause degradation of the protein membrane of phages. Previous studies have demonstrated that exposure to peroxide increases the rate of protein degradation by destabilizing proteins and increasing their susceptibility to proteolysis (Fligiel et al., Protein degradation following treatment with hydrogen peroxide. Am J Pathol 1984, 115(3), 418-25; Kocha et al., Hydrogen peroxide-mediated degradation of protein: different oxidation modes of copper- and iron-dependent hydroxyl radicals on the degradation of albumin. Biochim Biophys Acta 1997, 1337(2), 319-26). Salicylic acid is known for its protein binding ability (Lee et al., Protein binding of acetylsalicylic acid and salicylic acid in porcine and human serum. Vet Hum Toxicol 1995, 37(3), 224-5; Verbeeck and Cardinal, Plasma protein binding of salicylic acid, phenytoin, chlorpromazine, propranolol and pethidine using equilibrium dialysis and ultracentrifugation. Arzneimittelforschung 1985, 35(6), 903-6) and it is believed that its high affinity for the protein membrane of the capsid or the tail fibers renders the phage inviable.

[0018] Surprisingly, Propionibacterium acnes bacteriophage was found to be stable in compositions containing salicylic acid. See, for example, FIG. 15. Thus, salicylic acid has been shown to be well tolerated by the phage and is a suitable anti-acne agent for combination. In embodiments, the anti-keratolytic activity of salicylic acid complements the phage activity by allowing deeper penetration of the phage, thereby enhancing the bactericidal efficiency of the phage. In embodiments, the phage described herein may be combined with salicylic acid in compositions to prevent and treat acne.

[0019] Benzoyl peroxide is not suitable for combination with the tested phages, e.g., in formulations stored for more than a few days (see FIG. 16), but benzoyl peroxide can be used with the phage product as part of an anti-acne combination (e.g., kit). In an embodiment, benzoyl peroxide is the active ingredient in a cleanser, which is applied to the skin and washed off prior to application of the phage composition / formulation. In an embodiment, the antikeratolytic and transient antibacterial action of benzoyl peroxide complements the specific deeper selective killing of P. acnes by the bacteriophage.

[0020] In an embodiment, the Propionibacterium acnes bacteriophage and the anti-acne compound (such as salicylic acid and / or sulfur) are in a single composition that is administered topically to the skin of a subject. In an embodiment, a kit is provided that includes the Propionibacterium acnes bacteriophage and the anti-acne compound (e.g., in separate containers such as bottles). In an embodiment, the Propionibacterium acnes bacteriophage is in one composition and the anti-acne compound (such as benzoyl peroxide, salicylic acid and / or sulfur) is in another composition, each composition being administered topically to the skin of a subject. In an embodiment, the Propionibacterium acnes bacteriophage is administered to a subject, followed by the anti-acne compound. In an embodiment, the anti-acne compound is administered to a subject, followed by the Propionibacterium acnes bacteriophage. In an embodiment, the subject's face is washed during which the anti-acne compound and the Propionibacterium acnes bacteriophage (in either order) are topically administered to the subject's face.

[0021] In embodiments, the effective amount of an anti-acne compound (such as benzoyl peroxide, salicylic acid, or sulfur) when used in combination with a Propionibacterium acnes bacteriophage is less than that required when the anti-acne compound is used alone. In embodiments, the effective amount of an anti-acne compound (such as benzoyl peroxide, salicylic acid, or sulfur) when used in combination with a Propionibacterium acnes bacteriophage is less than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10% less than that required when the anti-acne compound is used alone.

[0022] definition The following definitions are included for purposes of understanding the present subject matter and for framing the appended claims: Abbreviations used herein have their conventional meaning within the chemical and biological arts.

[0023] While various embodiments and aspects of the invention have been illustrated and described herein, it will be apparent to those skilled in the art that such embodiments and aspects are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be used in practicing the invention.

[0024] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described. All documents or portions of documents cited in this application, including, but not limited to, patents, patent applications, articles, books, manuals, and papers, are hereby expressly incorporated by reference in their entirety for any purpose.

[0025] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.See, for example, Singleton et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY, 2nd ed., J.Wiley&Sons (New York, NY 1994); Sambrook et al., MOLECULAR CLONING, A LABORATORY MANUAL, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989).Any methods, devices and materials similar or equivalent to those described herein can be used in the practice of the present invention.The following definitions are provided to facilitate understanding of certain terms frequently used herein and are not intended to limit the scope of the present disclosure.

[0026] As used herein, a "Propionibacterium acnes bacteriophage" is a bacteriophage that infects, replicates within, and kills P. acnes cells. In an embodiment, the P. acnes bacteriophage is a lytic P. acnes bacteriophage. In an embodiment, the P. acnes bacteriophage is capable of lysing P. acnes bacteria and is unable to lyse any bacteria other than P. acnes. In an embodiment, the P. acnes bacteriophage is unable to maintain lysogeny within the bacterium. In an embodiment, the use of a bacteriophage that is able to lyse P. acnes but is unable to maintain lysogeny has the advantage that the bacteriophage cannot remain dormant within the bacterium, but does lyse and therefore kill the bacterium. In an embodiment, the P. acnes bacteriophage lacks the ability to express at least one gene required to maintain lysogeny. The term "lacking the ability to express at least one gene necessary to maintain lysogeny" is intended to indicate that the P. acnes bacteriophage lacks the ability to produce a fully functional protein product necessary to maintain lysogeny as a result of one or more point mutations or complete or partial deletions in the genome. In embodiments, the P. acnes bacteriophage has a genome lacking all or a portion of at least one gene necessary to maintain lysogeny (e.g., a strain is or is derived from a strain, whether artificial or natural, lacking all or a portion of at least one gene necessary to maintain lysogeny). However, in embodiments, the P. acnes bacteriophage may also contain defects (e.g., mutations, insertions or deletions) in the genome in non-coding regions that may affect the ability of the phage to maintain lysogeny, such as defects in genome integration sites (e.g., a / att / sites) or repressor binding sites. In embodiments, the P. acnes bacteriophage is naturally occurring and isolated, with the added advantage that no artificial mutations need to be introduced into the bacteriophage, hi embodiments, the P. acnes bacteriophage is capable of lysing multiple strains of P. acnes bacteria.In embodiments, the P. acnes bacteriophage is capable of lysing at least about 5, 10, 15, 20, 25, 30 or more strains of P. acnes bacteria. Non-limiting examples of P. acnes bacteriophages are disclosed herein. In embodiments, the P. acnes bacteriophage has a genome having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 95%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% sequence identity to SEQ ID NO:1. In embodiments, the P. acnes bacteriophage has a genome having or includes the sequence of SEQ ID NO:1. In an embodiment, the genome of the P. acnes bacteriophage has no insertions or deletions compared to SEQ ID NO: 1. In an embodiment, the genome of the P. acnes bacteriophage has no insertions or deletions compared to SEQ ID NO: 1, only conservative substitutions. In embodiments, the P. acnes bacteriophage is one of the following exemplary isolates of P. acnes bacteriophage deposited under the terms of the Budapest Treaty at the National Collection of Industrial, Marine and Food Bacteria (NCIMB), Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, United Kingdom, under the following accession numbers: accession number NCIMB 41332 (isolate PA6); accession number NCIMB 41334 (isolate 1874); accession number NCIMB 41333 (isolate 1878); accession number NCIMB 41335 (isolate 1905); accession number NCIMB 41349 (isolate 1894); accession number NCIMB 41350 (isolate 103609); accession number NCIMB 41351 (isolate 103672). In an embodiment, the host bacterium, a non-limiting example of which is P. acnes, is AT1, deposited as NCIMB 41336.In embodiments, the P. acnes bacteriophage has a genome having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 95% or 99% sequence identity to the genome of the bacteriophage deposited under accession number NCIMB 41349. In embodiments, the P. acnes bacteriophage has a genome having at least 87% sequence identity to the genome of the bacteriophage deposited under accession number NCIMB 41350. In embodiments, the P. acnes bacteriophage has a genome having at least 88% sequence identity to the genome of the bacteriophage deposited under accession number NCIMB 41351. Additional non-limiting description relating to P. acnes bacteriophages is provided in U.S. Patent No. 9,068,159 B2, issued June 30, 2015, the entire contents of which are incorporated herein by reference. The terms "phage" and "bacteriophage" are used interchangeably herein.

[0027] As used herein, "degrading" a biofilm means cleaving (e.g., by enzymatic activity) a covalent bond of at least one compound that forms part of the biofilm. Non-limiting examples of compounds that may form part of a biofilm include polymers, glycosides, proteins, polysaccharides, and nucleic acids. As used herein, a "P. acnes biofilm degrading enzyme" is an enzyme that degrades at least one compound that forms part of a P. acnes biofilm.

[0028] Enzymes provided herein include any naturally occurring form, homologue, isoform or variant that maintains enzymatic activity (within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to the native protein). In embodiments, variants have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 contiguous amino acid portion) compared to the naturally occurring form.

[0029] The term "isolated," as applied to a bacterium or bacteriophage, refers to a bacterium or bacteriophage that (1) has been separated from at least some of the components that accompanied it when it was originally produced (whether in nature or in an experimental setting) and / or (2) has been produced, prepared, purified, and / or manufactured by the hand of man using artificial culture conditions, such as, but not limited to, growth on plates and / or in a fermenter. Isolated bacteria include bacteria that are cultured, even if such culture is not a monoculture. In an embodiment, an isolated bacterium is a bacterium that is cultured as a monoculture (e.g., on plates or in a liquid culture such as a fermenter). Isolated bacteria and bacteriophages may be separated from at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 99% or more (e.g., by weight) of other components with which they were initially associated. In embodiments, isolated bacteria are greater than about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more (e.g., by weight) pure. In embodiments, the isolated bacteriophage is greater than about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or greater than about 99% pure (e.g., by weight). In embodiments, the compositions provided herein comprise one or more isolated bacteriophages. In embodiments, the compositions provided herein comprise an isolated bacteriophage. In embodiments, the administered bacteriophage is an isolated bacteriophage. In embodiments, the compositions provided herein comprise one or more isolated bacteria. In embodiments, the compositions provided herein comprise an isolated bacterium. In embodiments, the administered bacteriophage is an isolated bacteriophage.

[0030] A "control" sample or value is a sample that serves as a standard, usually a known standard, for comparison with a test sample. For example, a test sample can be taken from a test condition, for example, in the presence of a test compound (e.g., an enzyme) or phage, and compared to a sample taken under known conditions, such as in the absence of the test compound, phage, or bacteria (negative control) or in the presence of a known compound, phage, or bacteria (positive control). A control can also represent an average value collected from several tests or results. A person skilled in the art will recognize that controls can be designed for the evaluation of any number of parameters. For example, controls can be devised to compare the efficacy of treatments based on pharmacological data (e.g., half-life, degradation of biofilms or components thereof, or bacterial cell lysis) or treatment strategies (e.g., comparison of side effects). A person skilled in the art will understand which controls are useful in a given situation and can analyze data based on comparison with the control value. Controls also help determine the significance of the data. For example, if the values ​​of a given parameter are significantly different in the controls, the variation of the test samples will not be considered significant.

[0031] "Nucleic acid" refers to nucleotides (e.g., deoxyribonucleotides or ribonucleotides) and polymers thereof in single-, double-, or multi-stranded form, or their complements. The terms "polynucleotide", "oligonucleotide", "oligo" or the like refer to a linear sequence of nucleotides in the usual customary sense. Oligonucleotides are typically about 5, 6, 7, 8, 9, 10, 12, 15, 25, 30, 40, 50 or more nucleotides up to about 100 nucleotides in length. Polynucleotides are polymers of any length, including larger lengths, e.g., 200, 300, 500, 1000, 2000, 3000, 5000, 7000, 10000, 20000, 30000, 40000, etc. Polynucleotides and oligonucleotides generally contain phosphodiester bonds, but in some cases include nucleic acid analogs that may have alternative backbones, including, for example, phosphoramidate, phosphorothioate, dithiophosphate, or O-methyl phosphoramidite linkages (see Eckstein, Oligonucleotides and Analogues: A Practical Approach, Oxford University Press); as well as peptide nucleic acid backbones and linkages. Other analog nucleic acids include nucleic acids with cationic, nonionic and non-ribose backbones, including those described in U.S. Pat. Nos. 5,235,033 and 5,034,506, and Chapters 6 and 7, ASC Symposium Series 580, Carbohydrate Modifications in Antisense Research, Sanghui & Cook, eds. Nucleic acids containing one or more carbocyclic sugars are also included within one definition of nucleic acid. Modifications of the ribose-phosphate backbone may be made for various reasons, such as to increase the stability and half-life of such molecules in physiological environments or as probes on biochips. Mixtures of naturally occurring nucleic acids and analogs can be made; alternatively, mixtures of different nucleic acid analogs, and mixtures of naturally occurring nucleic acids and analogs may be made.

[0032] The term "bp" and the like refers in the ordinary and customary sense to the indicated number of base pairs.

[0033] "Percent sequence identity" is determined by comparing two optimally aligned sequences over a comparison window, where a portion of the comparison window of a polynucleotide or polypeptide sequence may contain additions or deletions (i.e., gaps) when compared to a reference sequence (which contains no additions or deletions) for optimal alignment of the two sequences. In an embodiment, the percentage is calculated by determining the number of positions where an identical nucleobase or amino acid residue is present in both sequences, dividing the number of matched positions by the total number of positions in the comparison window, and multiplying the result by 100 to obtain the percent sequence identity.

[0034] The term "identical" or percent "identity," in the context of two or more nucleic acid or polypeptide sequences, refers to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., for example, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identity over an entire nucleic acid or polypeptide sequence or a specified region of a particular portion or domain of a nucleic acid or polypeptide) when compared and aligned for optimal correspondence over a comparison window or designated region as measured using one of the sequence comparison algorithms below or by manual alignment and visual inspection. Such sequences are then said to be "substantially identical." This definition also extends to the complement of a test sequence. In embodiments, identity is about 20, 50, 100, 1000, 2500, 5000, 7500, 10000, 15000, 20000, 25000, or 30000 amino acids or nucleotides in length, or at least about 20, 50, 100, 1000, 2500, 5000, 7500, 10000, 15000, 20000, 25000, or 30000 amino acids or nucleotides in length. The identity exists over a region that is up to about 31000, 32000, 33000, 34000 or 35000 amino acids or nucleotides in length, less than about 31000, 32000, 33000, 34000 or 35000 amino acids or nucleotides in length, or at least about 31000, 32000, 33000, 34000 or 35000 amino acids or nucleotides in length. Optionally, the identity exists over a region that is at least about 10 to about 100, about 20 to about 75, about 30 to about 50 amino acids or nucleotides in length. Optionally, the identity exists over a region that is at least about 50 amino acids in length, or more preferably over a region that is 100 to 500 or 1000 or more amino acids in length. Phages that contain a nucleic acid (eg, a genome or portion thereof) having a sequence substantially identical to any of SEQ ID NOs:1, 11, 13, 15, 17, 19, 21, 23, 25, or 27 are included herein.Non-limiting examples of phages provided herein include those having a genome having a sequence substantially identical to SEQ ID NO:1.

[0035] In sequence comparison, typically one sequence serves as a reference sequence, to which test sequences are compared. When using sequence comparison algorithms, test and reference sequences are entered into a computer, subsequence coordinates are designated if necessary, and sequence algorithm program parameters are designated. Preferably, default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identity of the test sequence relative to the reference sequence based on the program parameters.

[0036] A "comparison window", as used herein, includes reference to any one segment of a number of contiguous positions over which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. In embodiments, the comparison window comprises about, or at least about, 20, 50, 100, 1000, 2500, 5000, 7500, 10000, 15000, 20000, 25000, or 30000 to about 31000, 32000, 33000, 34000, or 35000, or less than about, or at least about, 31000, 32000, 33000, 34000, or 35000 contiguous positions. In embodiments, the comparison window comprises about 20 or at least about 20 to about 31,000, less than about 31,000 or at least about 31,000 consecutive positions. In embodiments, the comparison window comprises about 25,000 or at least about 25,000 to about 31,000, less than about 31,000 or at least about 31,000 consecutive positions. In embodiments, the comparison window comprises about 26,000 or at least about 26,000 to about 31,000, less than about 31,000 or at least about 31,000 consecutive positions. In embodiments, the comparison window comprises about 27,000 or at least about 27,000 to about 31,000, less than about 31,000 or at least about 31,000 consecutive positions. In embodiments, the comparison window comprises about 28,000 or at least about 28,000 to about 31,000, less than about 31,000 or at least about 31,000 consecutive positions. In embodiments, the comparison window comprises about 29,000 or at least about 29,000 to about 31,000, less than about 31,000 or at least about 31,000 contiguous positions. In embodiments, the comparison window comprises about 30,000 or at least about 30,000 to about 31,000, less than about 31,000 or at least about 31,000 contiguous positions. In embodiments, the comparison window comprises about 20 to about 600, about 50 to about 200, or about 100 to about 150 contiguous positions. In embodiments, the comparison window is the full length of a reference sequence, such as the sequence of a bacteriophage genome. Methods for alignment of sequences for comparison are well known in the art.In embodiments, optimal alignment of sequences for comparison can be accomplished, for example, by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the similarity search method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by manual alignment and visual inspection (see, e.g., Current Protocols in Molecular Biology (Ausubel et al., eds. 1995 Supplement)).

[0037] Examples of algorithms suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., Nuc. Acids Res. 25:3389-3402 (1977) and Altschul et al., J. Mol. Biol. 215:403-410 (1990), respectively. As one skilled in the art will recognize, software for performing BLAST analysis is publicly available through the website of the National Center for Biotechnology Information (NCBI). In embodiments, BLAST and BLAST 2.0 are used to determine percent sequence identity for nucleic acids and proteins, with parameters as described herein. In embodiments, the BLAST algorithm involves first identifying high-scoring sequence pairs (HSPs) by identifying short words of length W in a query sequence that, when aligned with words of the same length in a database sequence, match or meet some positive threshold score T. In embodiments, T is referred to as the neighborhood word score threshold (Altschul et al., supra). In embodiments, these initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. In embodiments, the word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. In embodiments, the cumulative score is calculated using the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0), for nucleotide sequences. In embodiments, for amino acid sequences, a scoring matrix is ​​used to calculate the cumulative score. In embodiments, the extension of the word hits in each direction is stopped when the cumulative alignment score is reduced by an amount X from its maximum achieved value; when the cumulative score falls below zero due to the accumulation of one or more negative-scoring residue alignments; or when the end of either sequence is reached.In an embodiment, the BLAST algorithm parameters W, T and X determine the sensitivity and speed of the alignment. In an embodiment, the NCBI BLASTN or BLASTP program is used to align sequences. In an embodiment, the BLASTN or BLASTP program uses the defaults used by NCBI. In an embodiment, the BLASTN program (for nucleotide sequences) uses as defaults: word size (W) of 28; expectation threshold (E) of 10; maximum matches in the query range set to 0; match / mismatch scores of 1, -2; linear gap cost; a filter for low complexity regions used; and a mask only for the lookup table used. In an embodiment, the BLASTP program (for amino acid sequences) uses as defaults: a word size (W) of 3; an expectation threshold (E) of 10; maximum matches within the query range set to 0; the BLOSUM62 matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1992)); gap cost of extent: 11 and extension: 1; and a conditional composition score matrix adjustment.

[0038] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of a corresponding naturally occurring amino acid, as well as to naturally occurring and non-naturally occurring amino acid polymers.

[0039] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those that are subsequently modified, such as hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Amino acid analogs are compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an alpha carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, such as homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics are chemical compounds that have a structure that differs from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid.

[0040] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides may also be referred to by their commonly accepted one-letter codes.

[0041] "Conservatively modified variants" applies to both amino acid and nucleic acid sequences. With respect to a particular nucleic acid sequence, conservatively modified variants are nucleic acids that code for the same or essentially the same amino acid sequence, or essentially the same sequence if the nucleic acid does not code for an amino acid sequence. Due to the degeneracy of the genetic code, a large number of functionally identical nucleic acids code for any given protein. For example, the codons GCA, GCC, GCG, and GCU all code for the amino acid alanine. Thus, at all positions where alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are "silent variations," which are a type of conservatively modified variation. All nucleic acid sequences herein that code for a polypeptide also describe all possible silent variations of the nucleic acid. Those skilled in the art will recognize that each codon of a nucleic acid (except AUG, which is usually the only codon for methionine, and TGG, which is usually the only codon for tryptophan) can be altered to result in a functionally identical molecule. Accordingly, each silent variation of a nucleic acid which encodes a polypeptide is implicit in each described sequence with respect to the expression product, but not with respect to the actual probe sequence.

[0042] With respect to amino acid sequences, one of skill in the art will recognize that individual substitutions into a peptide, polypeptide, or protein sequence that change a single amino acid are "conservatively modified variants," in which the change replaces the amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to, and do not exclude, polymorphic variants, interspecies homologs, and alleles.

[0043] Each of the following eight groups contains amino acids that are conservative substitutions for one another: 1) alanine (A), glycine (G); 2) aspartic acid (D), glutamic acid (E); 3) asparagine (N), glutamine (Q); 4) arginine (R), lysine (K); 5) isoleucine (I), leucine (L), methionine (M), valine (V); 6) phenylalanine (F), tyrosine (Y), tryptophan (W); 7) serine (S), threonine (T); and 8) cysteine ​​(C), methionine (M) (see, e.g., Creighton, Proteins (1984)).

[0044] The term "disease" refers to any deviation from normal health in a mammal, including states when disease symptoms are present, as well as states where deviations (e.g., dysbiosis, infection, genetic mutation, genetic defect, etc.) have occurred but symptoms are not yet evident. In an embodiment, the disease is acne. In an embodiment, the disease includes skin dysbiosis. In an embodiment, the methods, compositions, systems, phages and probiotic bacteria provided herein are suitable for use in subjects that are members of the vertebrate class, mammals, including, but not limited to, primates (such as humans), livestock, working animals and pets (e.g., companion animals). In an embodiment, the subject is a human subject. As used herein, a "symptom" of a disease includes clinical or laboratory signs associated with the disease, but is not limited to those that can be felt or observed by the subject.

[0045] As used herein, the term "skin dysbiosis" refers to differences in the skin microbiome compared to a healthy or population. In embodiments, the dysbiosis is on the skin surface, in the skin (e.g., in an area of ​​skin or a layer of skin cells), in the glands, and / or in the pores of the skin. In embodiments, the dysbiosis is in sweat and / or sebum. In embodiments, the skin is on the face (e.g., on the subject's forehead, one or more cheeks, nose, or chin). In embodiments, the skin is on the shoulders, chest, or back. In embodiments, the skin dysbiosis includes changes in the microbiome symbiont species diversity compared to a healthy or population, and may include a decrease in beneficial microorganisms and / or an increase in pathogenic symbionts (pathogenic or potentially pathogenic microorganisms) and / or a decrease in overall microbiome species diversity. Many factors can result in dysbiosis, including hormonal changes (e.g., during puberty), infrequent washing, use of cosmetics, use of antibiotics, psychological and physical stress, radiation, and dietary changes.

[0046] In an embodiment, the composition is administered to a subject suffering from acne in a "therapeutically effective dose". The amount effective for this use may depend on the severity of the disease and the general state of the patient's health. Single or multiple administrations of the composition may be administered depending on the dosage and frequency required and tolerated by the patient. A "patient" or "subject" includes both humans and other animals, particularly mammals. Thus, the method is applicable to both human therapy and veterinary applications.

[0047] "Pharmaceutically acceptable excipients" and "pharmacologically acceptable carriers" refer to substances that aid in the administration and absorption of active agents by a subject and that may be included in the compositions of the present invention without causing significant adverse toxicological effects to the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline, lactated Ringer's solution, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavorings, saline solutions (such as Ringer's solution), alcohol, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethylcellulose, polyvinyl pyrrolidine, and pigments, and the like. Such preparations may be sterilized and, if desired, mixed with auxiliary substances such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring and / or aromatic substances and the like which do not deleteriously react with the bacteriophages, probiotic bacteria and / or compounds of the invention.Those skilled in the art will recognize that other pharmaceutical excipients are useful in the present invention.

[0048] The term "contacting" may include reacting, interacting, or physically contacting two species, where the two species may be, for example, an enzyme described herein and a biofilm containing a substrate for the enzyme. In another example, the two species may be a bacteriophage and a cell of a bacterial species that the bacteriophage infects. In an embodiment, contacting includes, for example, interacting a bacteriophage described herein with a P. acnes cell. In an embodiment, contacting includes, for example, interacting an enzyme described herein with a P. acnes biofilm.

[0049] A "patient" or "subject in need of treatment" is a living member of the animal kingdom who is suffering from or may suffer from the indicated disorder. In an embodiment, the subject is a member of a species that includes individuals who naturally suffer from the disease. In an embodiment, the subject is a mammal. Non-limiting examples of mammals include rodents (e.g., mice and rats), primates (e.g., lemurs, bush babies, monkeys, apes, and humans), rabbits, dogs (e.g., companion dogs, service dogs, or work dogs such as police dogs, military dogs, racing dogs, or show dogs), cats (e.g., house cats), livestock (pigs, cows, donkeys, mules, bison, goats, camels, and sheep), and deer. In an embodiment, the subject is a human.

[0050] The terms "subject," "patient," "individual," etc. are not intended to be limiting and are generally interchangeable, i.e., an "individual" described as a "patient" does not necessarily have a given disease, but may merely be seeking medical advice.

[0051] As used herein, the abbreviation "sp." for species refers to at least one species of the indicated genus (e.g., one, two, three, four, five or more species). The abbreviation "spp." for species refers to two or more species of the indicated genus (e.g., two, three, four, five, six, seven, eight, nine, ten or more). In embodiments, the methods and compositions provided herein include a single species within the indicated genus or genera, or two or more (e.g., a plurality including more than two) species within the indicated genus or genera. In embodiments, one, two, three, four, five or more, or all of the indicated species are isolated. In embodiments, the indicated species are administered together. In embodiments, each of the indicated species is present in a single composition comprising each of the species. In embodiments, each of the bacterial species are administered simultaneously, for example within about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 30 or 60, 1-5, 1-10, 1-30, 1-60 or 5-15 seconds or minutes of each other.

[0052] In this disclosure, "comprises," "comprising," "containing," and "having" and the like may have the meaning ascribed to these words in U.S. patent law and may mean "includes," "including," and the like. Thus, the transitional word "comprising," which is synonymous with "including," "containing," or "characterized by," is inclusive or open-ended and does not exclude additional, unrecited features, integers, steps, operations, elements, and / or components. "Consisting essentially of" or "consisting essentially" likewise has the meaning ascribed to it in U.S. patent law and the term is open-ended, permitting the presence of more than what is recited, but excluding prior art embodiments, so long as the basic or novel characteristics of the recited items are not altered by the presence of more than what is recited. In contrast, the transitional phrase "consisting of" excludes any features, integers, elements, steps, operations, components and / or ingredients not specified.

[0053] As used herein, the term "about" in the context of a numerical value or range means ±10% from the recited or claimed numerical value or range, unless the context requires a more specific range.

[0054] In the description and claims, phrases such as "at least one of" or "one or more of" may be present before a conjunctive list of elements or features. The term "and / or" may also be present in a list of two or more elements or features. Unless otherwise implicitly or explicitly contradicted by the context in which the phrase is used, such phrases are intended to mean any of the listed elements or features individually, or any of the listed elements or features in combination with any of the other listed elements or features. For example, the phrases "at least one of A and B," "one or more of A and B," and "A and / or B" are intended to mean "A alone, B alone, or A and B together," respectively. A similar interpretation is intended for lists containing three or more items. For example, the phrases "at least one of A, B, and C," "one or more of A, B, and C," and "A, B, and / or C" are intended to mean "A alone, B alone, C alone, A and B together, A and C together, B and C together, or A, B and C together." Furthermore, use of the term "based on" in the specification and claims is intended to mean "based at least in part on," allowing for unrecited features or elements.

[0055] When a parameter range is provided, it is understood that all integers and tenths thereof within the range are also provided by the present invention. For example, "0.2 to 5 mg" is a disclosure of 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, etc. up to and including 5.0 mg.

[0056] As used herein and throughout the claims which follow, the meanings of "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0057] As used herein, "treating" a condition, disease or disorder or a symptom associated with a condition, disease or disorder, or this "treatment" refers to an approach to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of the severity of a condition, disorder or disease, stabilization of the condition, disorder or disease state, prevention of the onset of a condition, disorder or disease, prevention of the spread of a condition, disorder or disease, delaying or slowing the progression of a condition, disorder or disease, delaying or slowing the onset of a condition, disorder or disease, amelioration or alleviation of a condition, disorder or disease state, whether partial or total. "Treating" can also mean inhibiting the progression of a condition, disorder or disease, slowing the progression of a condition, disorder or disease temporarily, but in some instances includes halting the progression of a condition, disorder or disease permanently. When treating acne, the term can refer to, for example, reducing the number or size of skin dysbiosis and / or cystic lesions, milia (closed blocked pores), blackheads (open blocked pores, oil exposed to air has a dark color, e.g., brown or black), mall reds, tender bumps (papules), pimples (pustules; papules with pus-tipped tips), large, hard, painful lumps (nodules) under the surface of the skin.

[0058] As used herein, the terms "treat" and "prevent" are not intended to be absolute terms. In embodiments, treatment can refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduction in the severity of the disease, condition or symptoms of the disease or condition suffered. In embodiments, a method for treating a disease is considered to be therapeutic if there is a 10% reduction in one or more symptoms of the disease in a subject compared to a control. Thus, the reduction can be 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or any percentage reduction between 10% and 100% when compared to a natural or control level. It is understood that treatment does not necessarily refer to a cure or complete ablation of a disease, condition or symptoms of a disease or condition. In embodiments, references to decrease, reduce, or inhibit include a change of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more when compared to a control level, and such terms can include, but do not necessarily include, complete elimination. Treatment can refer to any delay in onset, amelioration of symptoms, improvement in the patient's skin appearance, and the like. The effect of treatment can be compared to an individual or pool of individuals not receiving the treatment, or to the same patient at different times before or during treatment. In embodiments, the severity of the disease is reduced by at least 10%, for example, when compared to the individual before administration or to a control individual not receiving the treatment. In some aspects, the severity of the disease is reduced by at least 25%, 50%, 75%, 80%, or 90%, or in some cases is no longer detectable using standard diagnostic techniques. In embodiments, the treatment is effective to reduce at least one symptom of acne. In embodiments, the treatment is effective to reduce the level of pimples (pustules) on the face, forehead, chest, back and / or shoulders of the subject. In embodiments, the treatment is effective to reduce the level of milia (closed blocked pores) on the face, forehead, chest, back and / or shoulders of the subject. In embodiments, the treatment is effective to reduce the level of blackheads (open blocked pores) on the face, forehead, chest, back and / or shoulders of the subject.In embodiments, the treatment is effective to reduce the level of papules on the face, forehead, chest, back and / or shoulders of the subject. In embodiments, the treatment is effective to reduce the level of hard, painful lumps (nodules) under the surface of the skin on the face, forehead, chest, back and / or shoulders of the subject. In embodiments, the treatment is effective to reduce the level of cystic lesions on the face, forehead, chest, back and / or shoulders of the subject. In embodiments, the level (e.g., number) is reduced compared to before treatment began. In embodiments, the level (e.g., number) is reduced compared to a corresponding subject afflicted with acne but not receiving a treatment. In embodiments, the level (e.g., number) is reduced compared to a corresponding subject afflicted with acne but not receiving a treatment comprising a bacteriophage.

[0059] The terms "effective amount", "effective dose", "therapeutically effective amount", and the like refer to an amount of an agent sufficient to ameliorate a disorder as described herein. For example, for a given parameter, a therapeutically effective amount will exhibit at least a 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90% or at least a 100% increase or decrease. The therapeutic effect can also be expressed as a "fold" increase or decrease. For example, a therapeutically effective amount can have at least a 1.2-fold, 1.5-fold, 2-fold, 5-fold or more effect over a control.

[0060] The term "diagnosis" refers to the relative probability that a subject has a given metabolic disorder. Symptoms and diagnostic criteria are summarized herein. Similarly, the term "prognosis" refers to the relative probability that a certain future outcome may occur in a subject. For example, in the context of the present invention, prognosis can refer to the likelihood that an individual will develop acne. Prognosis can refer to the likely severity of a disease (e.g., severity of symptoms, rate of functional decline, etc.) As those skilled in the art of medical diagnostics will recognize, the terms are not intended to be absolute.

[0061] Compositions and Combinations Comprising Bacteriophages In aspects, provided herein are compositions comprising, consisting essentially of, or consisting of at least one P. acnes bacteriophage, at least one anti-acne compound, and a pharma- ceutically acceptable carrier.

[0062] In aspects, provided herein are compositions comprising at least one Propionibacterium acnes bacteriophage, no more than one anti-acne compound, and a pharma- ceutically acceptable carrier.

[0063] In an aspect, provided herein is a composition comprising an active ingredient consisting of at least one Propionibacterium acnes bacteriophage and no more than one anti-acne compound, and a pharma- ceutically acceptable carrier.

[0064] In aspects, provided herein are compositions comprising at least one P. acnes bacteriophage, at least one anti-acne compound, and a pharma- ceutically acceptable carrier, the compositions being free of probiotic bacteria.

[0065] In an embodiment, at least one anti-acne compound is benzoyl peroxide. In an embodiment, benzoyl peroxide is present in a concentration of 2.5% to 10% (weight / volume). In an embodiment, benzoyl peroxide is present in a concentration of less than 2.5% but greater than about 0.1%, 0.5%, 1%, 1.5% or 2% (weight / volume). In an embodiment, benzoyl peroxide is present in a concentration of 2.5% to 10%, for example, about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10% (weight / volume). In an embodiment, benzoyl peroxide is present in a concentration of less than 2.5% but greater than about 0.1%, 0.5%, 1%, 1.5% or 2% (weight / volume).

[0066] In an embodiment, at least one anti-acne compound is salicylic acid. In an embodiment, salicylic acid is present in a concentration of 0.5% to 2% (weight / volume). In an embodiment, salicylic acid is present in a concentration of less than 0.5% but greater than about 0.1% (weight / volume). In an embodiment, salicylic acid is present in a concentration of 0.5% to 2%, for example, about 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9% or 2% (weight / volume). In an embodiment, salicylic acid is present in a concentration of less than 0.5% but greater than about 0.1% (weight / volume).

[0067] In an embodiment, at least one anti-acne compound is sulfur. In an embodiment, sulfur is present in a concentration of 3% to 10% (weight / volume). In an embodiment, sulfur is present in a concentration of less than 3% but greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight / volume). In an embodiment, sulfur is present in a concentration of 3% to 10%, for example, about 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10% (weight / volume). In an embodiment, sulfur is present in a concentration of less than 3% but greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight / volume). In embodiments, resorcinol is present in a concentration of 2% and sulfur is present in a concentration of 3% to 8% (e.g., about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5% or 8%) (weight / volume).

[0068] In an embodiment, the at least one anti-acne compound is resorcinol and sulfur. In an embodiment, resorcinol is present at a concentration of 2% and sulfur is present at a concentration of 3% to 8% (weight / volume). In an embodiment, resorcinol is present at a concentration of 2% and sulfur is present at a concentration of 3% to 8% (e.g., about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, or 8%) (weight / volume).

[0069] In an embodiment, the at least one anti-acne compound comprises resorcinol monoacetate and sulfur. In an embodiment, the resorcinol monoacetate is present in a concentration of 3% and the sulfur is present in a concentration of 3% to 8% (weight / volume). In an embodiment, the resorcinol monoacetate is present in a concentration of 3% and the sulfur is present in a concentration of 3% to 8% (e.g., about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, or 8%) (weight / volume).

[0070] In an embodiment, the P. acnes bacteriophage is about 1×10 6 , 2×10 6 , 3×10 6 , 4×10 6 , 5×10 6 , 6×10 6 , 7×10 6 , 8×10 6 , 9×10 6 , 1×10 7 , 2×10 7 , 3×10 7 , 4×10 7 , 5×10 7 6×10 7 , 7×10 7 , 8×10 7 , 9×10 7 , 1×10 8 , 2×10 8 , 3×10 8 , 4×10 8 , 5×10 8 , 6×10 8 , 7×10 8 , 8×10 8 , 9×10 8 , 1×10 9 , 2×10 9 , 3×10 9 , 4×10 9 , 5×10 9 , 6×10 9 , 7×10 9 , 8×10 9 , 9×10 9 , 1×10 10 , 2×1010 , 3×10 10 , 4×10 10 , 5×10 10 , 6×10 10 , 7×10 10 , 8×10 10 , 9×10 10 , or 1 × 10 11 In an embodiment, the P. acnes bacteriophage is present in an amount of about 1×10 6 ~1×10 11 In an embodiment, the P. acnes bacteriophage is present in an amount of about 1×10 6 ~1×10 8 , about 1×10 8 ~1×10 9 , about 1×10 9 ~1×10 10 , about 1×10 9 ~1×10 11 Or about 1 x 10 10 ~1×10 11 It is present in amounts of pfu.

[0071] In an embodiment, the probiotic bacteria is about 1×10 6 , 2×10 6 , 3×10 6 , 4×10 6 , 5×10 6 , 6×10 6 , 7×10 6 , 8×10 6 , 9×10 6 , 1×10 7 , 2×10 7 , 3×10 7 , 4×10 7 , 5×10 7 6×10 7 , 7×10 7 , 8×10 7 , 9×10 7 , 1×10 8 , 2×10 8 , 3×10 8 , 4×10 8 , 5×10 8 , 6×10 8 , 7×10 8 , 8×108 , 9×10 8 , 1×10 9 , 2×10 9 , 3×10 9 , 4×10 9 , 5×10 9 , 6×10 9 , 7×10 9 , 8×10 9 , 9×10 9 , 1×10 10 , 2×10 10 , 3×10 10 , 4×10 10 , 5×10 10 , 6×10 10 , 7×10 10 , 8×10 10 , 9×10 10 , or 1 × 10 11 In an embodiment, the probiotic bacteria is present in an amount of about 1×10 6 ~1×10 11 In an embodiment, the probiotic bacteria is present in an amount of about 1×10 6 ~1×10 8 , about 1×10 8 ~1×10 9 , about 1×10 9 ~1×10 10 , about 1×10 9 ~1×10 11 Or about 1 x 10 10 ~1×10 11 It is present in amounts of cfu.

[0072] In embodiments, the anti-acne compound is an antibiotic, a retinoid, or an alpha hydroxy acid.

[0073] In an aspect, provided herein is a composition comprising a P. acnes bacteriophage and an enzyme.

[0074] In embodiments, provided herein are combinations comprising, consisting essentially of, or consisting of at least one P. acnes bacteriophage, at least one anti-acne compound, wherein each of the at least one P. acnes bacteriophage and the at least one anti-acne compound is in a composition further comprising a pharma- ceutically acceptable carrier.

[0075] In an aspect, provided herein is a combination comprising a P. acnes bacteriophage and an enzyme.

[0076] In an embodiment, the P. acnes bacteriophage has a linear, double-stranded DNA genome.

[0077] In an embodiment, the P. acnes bacteriophage is within the bacteriophage family Siphoviridae.

[0078] In an embodiment, the bacteriophage is a wild-type bacteriophage. In an embodiment, the bacteriophage has a genome with a nucleotide sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical to the genomic sequence of a wild-type acnes bacteriophage. A non-limiting example of a genomic sequence of a wild-type acnes bacteriophage is as follows:

[0079] [ka] TIFF2024099592000003.tif218162TIFF2024099592000004.tif217162TIFF2024099592000005.tif218162TIFF2024099592000006.tif218162TIFF2024099592000007.tif218162TIFF2024099592000008.tif219162TIFF2024099592000009.tif217161TIFF2024099592000010.tif218164TIFF2024099592000011.tif218163TIFF2024099592000012.tif218163TIFF2024099592000013.tif218160TIFF2024099592000014.tif218161TIFF2024099592000015.tif217159TIFF2024099592000016.tif216161TIFF2024099592000017.tif32159

[0080] In embodiments, the bacteriophage is a bacteriophage deposited under accession number NCIMB 41349, 41350 or 41351. In embodiments, the bacteriophage has a genome with a nucleotide sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical to the genome of the bacteriophage deposited under accession number NCIMB 41349. In embodiments, the bacteriophage has a genome with a nucleotide sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical to the genome of the bacteriophage deposited under accession number NCIMB 41350. In embodiments, the bacteriophage has a genome with a nucleotide sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical to the genome of the bacteriophage deposited under accession number NCIMB 41351.

[0081] In embodiments, the bacteriophage has a genome having a nucleotide sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical to the nucleotide sequence of SEQ ID NO: 1. In embodiments, the bacteriophage has a genome having a nucleotide sequence that is identical to the nucleotide sequence of SEQ ID NO:1.

[0082] In an embodiment, the genome of the bacteriophage includes, from the 5' to 3' end, a small terminase, a large terminase, a portal protein, gp4, a scaffold protein, a major head protein, gp7, gp8, gp9, gp10, a major tail protein, gp12, gp13, a tape measure protein, a minor tail subunit, and optionally a protease, gp17, gp18, a tail protein, and a nucleotide sequence. These proteins encode the amidase, holin, gp22, gp23, sigma factor, gp25, gp26, gp27, gp28, gp29, gp30, DNA primase, DNA primase 2, gp33, DNA helicase, gp35, gp36, exonuclease, p38, gp39, gp40, gp41, gp42, gp43, gp44, gp45, gp46, gp47, and gp48.

[0083] In an embodiment, the composition further comprises a P. acnes biofilm degrading enzyme.

[0084] In an embodiment, the enzyme is an anti-aging enzyme. In an embodiment, the anti-aging enzyme is a superoxide dismutase or a peroxidase.

[0085] In an embodiment, the enzyme is a P. acnes biofilm degrading enzyme. In an embodiment, the enzyme is a glycosidase, a protease, a deoxyribonuclease, or a restriction endonuclease. In an embodiment, the enzyme is a glycosidase. In an embodiment, the glycosidase is a glycoside hydrolase. In an embodiment, the enzyme catalyzes the hydrolysis of linear polymers of N-acetyl-D-glucosamine. In an embodiment, the enzyme is a β-hexosaminidase. In an embodiment, the enzyme hydrolyzes β-1,6-glycosidic bonds in acetylglucosamine polymers. In an embodiment, the enzyme is a DNase I, a restriction endonuclease, papain, bromelain, trypsin, proteinase K, subtilisin, serratiopeptidase, dispersin, alginate lyase, amylase, or cellulase. In an embodiment, the enzyme is dispersin B. In an embodiment, the enzyme is a protease, the protease being proteinase K or subtilisin.

[0086] In an embodiment, the enzyme is dispersin. In an embodiment, the enzyme is dispersin B. In an embodiment, the enzyme is a naturally occurring form, homologue, isoform or variant of dispersin (such as dispersin B) that maintains enzymatic activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to the native protein). In an embodiment, the variant has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a portion of 50, 100, 150 or 200 consecutive amino acids) compared to the naturally occurring form. Non-limiting examples of DNA sequences encoding dispersin B are as follows:

[0087] [ka] TIFF2024099592000019.tif106163

[0088] A non-limiting example of a dispersin B amino acid sequence is as follows:

[0089] [ka]

[0090] In an embodiment, the enzyme is alginate lyase. In an embodiment, the enzyme is a naturally occurring form, homologue, isoform or variant of alginate lyase that maintains the enzymatic activity of alginate lyase (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to the native protein). In an embodiment, the variant has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a portion of 50, 100, 150 or 200 consecutive amino acids) compared to the naturally occurring form. Non-limiting examples of DNA sequences encoding alginate lyase are as follows:

[0091] [ka]

[0092] A non-limiting example of an alginate lyase amino acid sequence is as follows:

[0093] [ka]

[0094] In an embodiment, the enzyme is an amylase. In an embodiment, the enzyme is a naturally occurring form, homologue, isoform or variant of an amylase that maintains the enzymatic activity of the amylase (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity of the native protein). In an embodiment, the variant has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a portion of 50, 100, 150 or 200 consecutive amino acids) to the naturally occurring form. Non-limiting examples of DNA sequences encoding amylases are as follows:

[0095] [ka] TIFF2024099592000024.tif74160

[0096] Non-limiting examples of amylase amino acid sequences are as follows:

[0097] [ka]

[0098] In an embodiment, the enzyme is a cellulase. In an embodiment, the enzyme is a naturally occurring form, homologue, isoform, or variant of cellulase that maintains the enzymatic activity of cellulase (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to the native protein). In an embodiment, the variant has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a portion of 50, 100, 150 or 200 consecutive amino acids) compared to the naturally occurring form. Non-limiting examples of DNA sequences encoding cellulases are as follows:

[0099] [ka]

[0100] Non-limiting examples of cellulase amino acid sequences are as follows:

[0101] [ka]

[0102] In an embodiment, the enzyme is proteinase K. In an embodiment, the enzyme is a naturally occurring form, homologue, isoform or variant of proteinase K that maintains the enzymatic activity of proteinase K (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to the native protein). In an embodiment, the variant has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a portion of 50, 100, 150 or 200 contiguous amino acids) compared to the naturally occurring form. Non-limiting examples of DNA sequences encoding proteinase K are as follows:

[0103] [ka]

[0104] A non-limiting example of a Proteinase K amino acid sequence is as follows:

[0105] [ka]

[0106] In an embodiment, the enzyme is subtilisin. In an embodiment, the enzyme is a naturally occurring form, homologue, isoform or variant of subtilisin that maintains the enzymatic activity of subtilisin (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity of the native protein). In an embodiment, the variant has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a portion of 50, 100, 150 or 200 contiguous amino acids) to the naturally occurring form. Non-limiting examples of DNA sequences encoding subtilisin are as follows:

[0107] [ka]

[0108] A non-limiting example of a subtilisin amino acid sequence is as follows:

[0109] [ka]

[0110] In an embodiment, the enzyme is trypsin. In an embodiment, the enzyme is a naturally occurring form, homologue, isoform or variant of trypsin that maintains the enzymatic activity of trypsin (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to the native protein). In an embodiment, the variant has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a portion of 50, 100, 150 or 200 consecutive amino acids) compared to the naturally occurring form. Non-limiting examples of DNA sequences encoding trypsin are as follows:

[0111] [ka]

[0112] A non-limiting example of a trypsin amino acid sequence is as follows:

[0113] [ka]

[0114] In an embodiment, the enzyme is serratiopeptidase. In an embodiment, the enzyme is a naturally occurring form, homologue, isoform or variant of serratiopeptidase that maintains the enzymatic activity of serratiopeptidase (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity of the native protein). In an embodiment, the variant has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a portion of 50, 100, 150 or 200 contiguous amino acids) to the naturally occurring form. Non-limiting examples of DNA sequences encoding serratiopeptidase are as follows:

[0115] [ka] TIFF2024099592000035.tif127161

[0116] A non-limiting example of a serratiopeptidase amino acid sequence is as follows:

[0117] [ka]

[0118] In an embodiment, the enzyme is a deoxyribonuclease. In an embodiment, the enzyme is a naturally occurring form, homologue, isoform, or variant of deoxyribonuclease that maintains the enzymatic activity of deoxyribonuclease (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to the native protein). In an embodiment, the variant has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 contiguous amino acid portion) compared to the naturally occurring form. In an embodiment, the enzyme is deoxyribonuclease I. In an embodiment, the deoxyribonuclease I is bovine pancreatic deoxyribonuclease I. A non-limiting example of a DNA sequence encoding bovine pancreatic desoxyribonuclease I is as follows:

[0119] [ka]

[0120] A non-limiting example of a bovine pancreatic desoxyribonuclease I amino acid sequence is as follows:

[0121] [ka]

[0122] In an embodiment, the composition or combination comprises probiotic bacteria.

[0123] In an embodiment, the probiotic bacteria are P. sp., Staphylococcus sp. and / or Corynebacterium sp. bacteria.

[0124] In an embodiment, the probiotic bacteria are bacteria within the class Betaproteobacteria.

[0125] In an embodiment, the probiotic bacteria is probiotic P. acnes.

[0126] In an embodiment, the P. acnes bacteria has (a) a 16S ribosomal DNA (rDNA) sequence with a T992C mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (b) a 16S rDNA sequence with a T838C mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (c) a 16S rDNA sequence with a C1322T mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (d) a 16S rDNA sequence with a C986T mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (e) a 16S rDNA sequence identical to the sequence of SEQ ID NO:3; and (f) a 16S rDNA sequence identical to the sequence of SEQ ID NO:4. (g) containing no linear plasmids; (h) containing no plasmids containing virulence factors; and / or (i) being free of plasmids encoding extrachromosomal lipases and / or adhesive virulence factors.

[0127] In embodiments, the P. acnes bacteria (a) produces less than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of the level of lipase produced by a pathogenic P. acnes strain when grown in planktonic culture; (b) produces less than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of the level of lipase produced by a pathogenic P. acnes strain when grown in adherent culture; (c) produce less than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of the level of lipase produced by pathogenic P. acnes strains; (d) adhere to epithelial cells at least 50% less than pathogenic P. acnes strains; and / or (e) are less inflammatory than pathogenic P. acnes strains.

[0128] In an embodiment, the combination or composition comprises at least one additional probiotic bacterium, hi an embodiment, the at least one additional probiotic bacterium comprises Propionibacterium granulosum and / or Propionibacterium avidum.

[0129] In an embodiment, the pathogenic P. acnes strain has (a) a 16S rDNA sequence with a G1058C mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (b) a 16S rDNA sequence with G1058C and A1201C compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (c) a 16S rDNA sequence with a G529A mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (d) a 16S rDNA sequence with G1004A and T1007C mutations compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (e) a 16S rDNA sequence with a G1268A mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2. (f) having a 16S rDNA sequence with T554C and G1058C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (g) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:5; (h) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:6; (i) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:7; (j) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:8; (k) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:9; and / or (l) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:10.

[0130] In an embodiment, the combination or composition further comprises at least one additional P. acnes bacteriophage.

[0131] In an embodiment, the combination or composition further comprises a pharma- ceutically acceptable carrier. In an embodiment, the pharma- ceutically acceptable carrier comprises an emulsion. In an embodiment, the emulsion is an oil-in-water emulsion or a water-in-oil emulsion. In an embodiment, the combination or composition comprises or is in the form of a cream, lotion, suspension, or aqueous solution.

[0132] In an embodiment, a composition comprising a bacteriophage is provided. In an embodiment, the composition is formulated for topical application to the skin (i.e., the composition is a topical composition). In an embodiment, the composition is a pharmaceutical composition.

[0133] In an aspect, a pharmaceutical composition is provided comprising a wild-type P. acnes bacteriophage and an isolated probiotic P. acnes bacterium. In an embodiment, the composition further comprises a pharma- ceutically acceptable carrier.

[0134] In an aspect, a pharmaceutical composition is provided comprising a bacteriophage and / or an isolated probiotic P. acnes bacteria and a pharma- ceutically acceptable carrier.

[0135] In an embodiment, the pharmaceutical composition is formulated for topical administration to the skin. In an embodiment, the pharma- ceutically acceptable carrier comprises an emulsion. In an embodiment, the emulsion is an oil-in-water emulsion or a water-in-oil emulsion. In an embodiment, the pharmaceutical composition is in the form of a cream, lotion, suspension or aqueous solution.

[0136] In embodiments, the composition or combination comprises at least about 2, 3, 4, 5, 6, 7, 8, 9, or 10 P. acnes bacteriophages. In embodiments, the P. acnes bacteriophage comprises more than one type of P. acnes bacteriophage.

[0137] In an embodiment, the combination or composition comprising isolated probiotic P. acnes bacteria may further comprise at least one additional bacterium.

[0138] In an embodiment, the P. acnes has a 16S rDNA sequence comprising T992C, T838C, C1322T and / or C986T mutations compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO: 2. In an embodiment, the P. acnes comprises a 16S rDNA sequence with T838C and C1322T mutations compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO: 2. In an embodiment, the P. acnes is a ProI strain. In an embodiment, the P. acnes comprises a 16S rDNA sequence with C986T and T992C mutations compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO: 2. In an embodiment, the P. acnes is a ProII strain. In an embodiment, the P. acnes bacteria (a) comprises a 16S rDNA sequence having a T992C mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (b) comprises a 16S rDNA sequence having a T838C mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (c) comprises a 16S rDNA sequence having a C1322T mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (d) comprises a 16S rDNA sequence having a C986T mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (e) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO:3; (f) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO:4. (g) does not contain a linear plasmid; (h) does not contain a plasmid containing a virulence factor; and / or (i) does not contain a plasmid encoding an extrachromosomal lipase and / or an adhesive virulence factor. In an embodiment, the P. acnes bacteria has a 16S rDNA identical to the sequence of SEQ ID NO: 3 or 4.

[0139] In embodiments, the P. acnes bacteria (a) produces less than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of the level of lipase produced by a pathogenic P. acnes strain when grown in planktonic culture; (b) produces less than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of the level of lipase produced by a pathogenic P. acnes strain when grown in adherent culture; (c) produce less than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of the level of lipase produced by pathogenic P. acnes strains; (d) adhere to epithelial cells at least 50% less than pathogenic P. acnes strains; and / or (e) are less inflammatory than pathogenic P. acnes strains. In an embodiment, the pathogenic P. acnes strain has (a) a 16S rDNA sequence with a G1058C mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (b) a 16S rDNA sequence with G1058C and A1201C compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (c) a 16S rDNA sequence with a G529A mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (d) a 16S rDNA sequence with G1004A and T1007C mutations compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (e) a 16S rDNA sequence with a G1268A mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2. (f) having a 16S rDNA sequence with T554C and G1058C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (g) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:5; (h) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:6; (i) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:7; (j) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:8; (k) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:9; and / or (l) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:10.

[0140] SEQ ID NO:2 is the 16S rDNA sequence representing the KPA171202 type strain and is as follows:

[0141] [ka]

[0142] SEQ ID NO:3 is the 16S rDNA sequence representing the ProI probiotic strain, which is as follows: Nucleotides 838..838 ProI mutation T838C Nucleotides 1322..1322 ProI mutation C1322T

[0143] [ka]

[0144] SEQ ID NO: 4 is the 16S rDNA sequence representing the ProII probiotic strain, which is as follows: Nucleotides 986..986 ProII mutation C986T Nucleotide 992..992 ProII mutation T992C

[0145] [ka]

[0146] In embodiments, the P. acnes bacteria produces less than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of the level of lipase produced by a pathogenic P. acnes strain when grown in suspension culture. In embodiments, the P. acnes bacteria produces about 1-5%, 1-10%, 1-20%, 1-30%, 5-50%, 5-40%, 5-30%, 5-20%, 5-10%, 10-50%, 10-40%, 10-30%, 10-20%, 20-50%, 20-40%, or 20-30% of the level of lipase produced by pathogenic P. acnes strains when grown in suspension culture. In embodiments, the P. acnes bacteria produces less than about 5% of the level of lipase produced by pathogenic P. acnes strains when grown in suspension culture. In embodiments, the P. acnes bacteria produces less than about 10% of the level of lipase produced by pathogenic P. acnes strains when grown in suspension culture. In embodiments, the P. acnes strains produce less than about 20% of the levels of lipase produced by pathogenic P. acnes strains when grown in suspension culture. In embodiments, the P. acnes strains produce less than about 30% of the levels of lipase produced by pathogenic P. acnes strains when grown in suspension culture. In embodiments, the P. acnes strains produce less than about 40% of the levels of lipase produced by pathogenic P. acnes strains when grown in suspension culture. In embodiments, the P. acnes strains produce less than about 50% of the levels of lipase produced by pathogenic P. acnes strains when grown in suspension culture. In embodiments, the P. acnes bacteria produces less than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of the levels of lipase produced by pathogenic P. acnes strains when grown in adherent culture. In embodiments, the P. acnes bacteria produces less than about 5% of the levels of lipase produced by pathogenic P. acnes strains when grown in adherent culture.In embodiments, P. acnes produces less than about 10% of the levels of lipase produced by pathogenic P. acnes strains when grown in adherent culture. In embodiments, P. acnes produces less than about 20% of the levels of lipase produced by pathogenic P. acnes strains when grown in adherent culture. In embodiments, P. acnes produces less than about 30% of the levels of lipase produced by pathogenic P. acnes strains when grown in adherent culture. In embodiments, P. acnes produces less than about 40% of the levels of lipase produced by pathogenic P. acnes strains when grown in adherent culture. In embodiments, P. acnes produces less than about 50% of the levels of lipase produced by pathogenic P. acnes strains when grown in adherent culture. In embodiments, the P. acnes bacteria produces about 1-5%, 1-10%, 1-20%, 1-30%, 5-50%, 5-40%, 5-30%, 5-20%, 5-10%, 10-50%, 10-40%, 10-30%, 10-20%, 20-50%, 20-40%, or 20-30% of the level of lipase produced by pathogenic P. acnes strains when grown in adherent culture. In embodiments, the lipase is an extracellular lipase.

[0147] In an embodiment, the level of lipase produced by P. acnes (e.g., probiotic or pathogenic P. acnes, for comparison) is the level of lipase in the culture supernatant. In an embodiment, the culture supernatant is filtered. In an embodiment, the culture supernatant is from a liquid (planktonic) culture. In an embodiment, the culture supernatant is from an adherent culture. Non-limiting examples of methods for detecting the level of lipase include absorbance, Bradford protein assay, biuret test derived assays, fluorescamine, amino black, colloidal gold, nitrogen detection, high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC / MS), enzyme-linked immunosorbent assay (ELISA), protein immunoprecipitation, immunoelectrophoresis, and Western blot.

[0148] In embodiments, P. acnes adheres to epithelial cells at least about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% less than pathogenic P. acnes strains. In embodiments, P. acnes adheres to epithelial cells at least about 50% less than pathogenic P. acnes strains. In embodiments, P. acnes adheres to epithelial cells at least about 60% less than pathogenic P. acnes strains. In embodiments, P. acnes adheres to epithelial cells at least about 70% less than pathogenic P. acnes strains. In embodiments, P. acnes adheres to epithelial cells at least about 80% less than pathogenic P. acnes strains. In embodiments, the P. acnes bacteria adhere to epithelial cells at least about 90% less than pathogenic P. acnes strains, in embodiments, the P. acnes bacteria adhere to epithelial cells 1-5%, 1-10%, 1-20%, 1-30%, 5-50%, 5-40%, 5-30%, 5-20%, 5-10%, 10-50%, 10-40%, 10-30%, 10-20%, 20-50%, 20-40%, or 20-30%, 50-60, 50-70, 50-80, 50-90, 60-80, or 70-90 less than pathogenic P. acnes strains.

[0149] In an embodiment, adhesion of P. acnes (e.g., probiotic or pathogenic P. acnes, for comparison) to epithelial cells is determined using A-432 epithelial cells. In an embodiment, the epithelial cells are confluent in tissue culture plates or flasks. In an embodiment, adhesion is detected by determining the number of colonies formed by P. acnes adhering to the cultured epithelial cells.

[0150] In an embodiment, the P. acnes bacteria is less inflammatory than a pathogenic P. acnes strain.

[0151] In an embodiment, the P. acnes strain is less inflammatory than a pathogenic P. acnes strain if a lower level of inflammatory cytokines (e.g., at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% less) is released by immune cells in contact with the P. acnes strain or a compound produced by the P. acnes strain compared to a pathogenic P. acnes strain or a compound produced by the bacteria of the pathogenic P. acnes strain. In an embodiment, the P. acnes strain is less inflammatory than a pathogenic P. acnes strain if a lower level of inflammatory cytokines is released in a tissue (such as skin tissue) in contact with the P. acnes strain. In an embodiment, the tissue is skin tissue. In an embodiment, the tissue is, for example, mouse ear tissue. In an embodiment, the inflammatory cytokine is IL-1β, IL-6, IL-17, or TNFα, or any combination thereof.

[0152] In an embodiment, the pathogenic P. acnes strain has (a) a 16S rDNA sequence with a G1058C mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (b) a 16S rDNA sequence with G1058C and A1201C compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (c) a 16S rDNA sequence with a G529A mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (d) a 16S rDNA sequence with G1004A and T1007C mutations compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (e) a 16S rDNA sequence with a G1268A mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2. (f) having a 16S rDNA sequence with T554C and G1058C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (g) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:5; (h) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:6; (i) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:7; (j) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:8; (k) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:9; and / or (l) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:10.

[0153] SEQ ID NO:5 is as follows (mutations compared to the 16SA sequence of reference strain KPA171202 are underlined):

[0154] [ka] TIFF2024099592000043.tif145162

[0155] SEQ ID NO:6 is as follows (mutations compared to the 16S sequence of reference strain KPA171202 are underlined):

[0156] [ka] TIFF2024099592000045.tif145159

[0157] SEQ ID NO:7 is as follows (mutations compared to the 16S sequence of reference strain KPA171202 are underlined):

[0158] [ka] TIFF2024099592000047.tif144160

[0159] SEQ ID NO:8 is as follows (mutations compared to the 16S sequence of reference strain KPA171202 are underlined):

[0160] [ka] TIFF2024099592000049.tif144160

[0161] SEQ ID NO:9 is as follows (mutations compared to the 16S sequence of reference strain KPA171202 are underlined):

[0162] [ka] TIFF2024099592000051.tif143161

[0163] SEQ ID NO:10 is as follows (mutations compared to the 16S sequence of reference strain KPA171202 are underlined):

[0164] [ka] TIFF2024099592000053.tif143161

[0165] In an embodiment, the at least one additional bacterium comprises, consists essentially of, or consists of probiotic bacteria. In an embodiment, the at least one bacterium comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains and / or species, less than about 10, 9, 8, 7, 6, 5, 4, 3, or 2 bacterial strains and / or species, or 1-10, 2-10, 3-10, 4-10, 5-10, 1-5, 2-5, 3-5, or 4-5 bacterial strains and / or species. In an embodiment, the at least one bacterium comprises a plurality of bacterial strains and / or species, for example at least about 2, 3, 4, 5, 6, 7, 8, 9, 10 bacterial strains and / or species. In an embodiment, the at least one bacterium comprises an isolated Propionibacterium granulosum bacterium, an isolated Propionibacterium avidum bacterium, an isolated Staphylococcus epidermidis bacterium, an isolated Staphylococcus aureus bacterium, and / or an isolated Corynebacterium jeikeium bacterium. In an embodiment, the at least one bacterium comprises one, two (in any combination), three (in any combination), four (in any combination), or five of the isolated Propionibacterium granulosum bacterium, isolated Propionibacterium avidum bacterium, isolated Staphylococcus epidermidis bacterium, isolated Staphylococcus aureus bacterium, and / or isolated Corynebacterium jeikeium bacterium.

[0166] In embodiments, the compositions or combinations provided herein include a enhancing peptide or enzyme that has one or more or any combination of the following properties: biofilm degradation, improved skin penetration, antibacterial, reduced inflammation (e.g., of the skin), reduced irritation (e.g., of the skin), reduced redness (e.g., of the skin), skin tightening, wrinkle removal, fine line removal, or other improved appearance (e.g., of the skin).

[0167] In an aspect, a composition is provided that includes a P. acnes bacteriophage and an anti-acne compound. In an embodiment, the composition includes a pharma- ceutically acceptable carrier. In an embodiment, the dose of the P. acnes bacteriophage is adjusted (e.g., increased or decreased) for stability. In an embodiment, the dose of the P. acnes bacteriophage is adjusted up or down depending on the anti-acne compound for its stability in combination with the anti-acne compound.

[0168] In an embodiment, a combination or system is provided that includes a P. acnes bacteriophage and one or more anti-acne compounds. In an example, the bacteriophage is in one composition (e.g., in one container, such as a bottle, tube, or other container) and the one or more anti-acne compounds are in another composition (e.g., in another container, such as a bottle, tube, or other container). In an embodiment, the composition that includes the bacteriophage includes a pharma- ceutically acceptable carrier. In an embodiment, the composition that includes the anti-acne compounds includes a pharma- ceutically acceptable carrier. In an embodiment, one or more additional compounds (e.g., an enzyme, a hydrating compound, an ultraviolet absorbing or blocking compound, etc.) are present in the composition that includes the bacteriophage, the composition that includes one or more anti-acne compounds, or a third separate composition (e.g., in a third container, such as a bottle, tube, or other container). In an embodiment, one or more probiotic bacteria are present in the composition that includes the bacteriophage, the composition that includes one or more anti-acne compounds, or a third separate composition (e.g., in a third container, such as a bottle, tube, or other container). In an embodiment, the combination or system further comprises instructions for administration.In an embodiment, the combination or system comprises at least about 2, 3, 4, 5, 6, 7, 8, 9, or 10 P. acnes bacteriophages.

[0169] In an embodiment, a combination or system is provided that includes a P. acnes bacteriophage and one or more probiotic bacteria and / or one or more compounds (such as one or more enzymes or anti-acne compounds). In an example, the bacteriophage is in one composition (e.g., in one container, such as a bottle, tube, or other container), the one or more probiotic bacteria are in another composition (in another container, such as a bottle, tube, or other container), and optionally, the additional one or more compounds are present in the composition that includes the bacteriophage, the composition that includes the one or more probiotic bacteria, or a third separate composition (in a third container, such as a bottle, tube, or other container). In an embodiment, the combination or system further includes instructions for administration. In an embodiment, the combination or system includes at least about 2, 3, 4, 5, 6, 7, 8, 9, or 10 P. acnes bacteriophages.

[0170] In an embodiment, the system, combination or composition includes an enzyme such as a biofilm degrading enzyme or an anti-aging enzyme. Non-limiting examples of biofilm degrading enzymes include DNase (e.g., DNase I), protease (e.g., papain, bromelain, trypsin, proteinase K, subtilisin or serratiopeptidase), glycosidase (e.g., dispersin, alginate lyase, amylase or cellulase). Non-limiting examples of anti-aging enzymes include superoxide dismutase and peroxidase.

[0171] In an embodiment, the system, combination, or composition includes a topical retinoid, an antibiotic, and / or an alpha hydroxy acid. In an embodiment, the system or composition further includes a topical retinoid. In an embodiment, the system or composition further includes an antibiotic. In an embodiment, the system or composition further includes an alpha hydroxy acid. In an embodiment, the system or composition further includes benzoyl peroxide, salicylic acid, sulfur, resorcinol, resorcinol monoacetate, or any combination thereof. In an embodiment, the benzoyl peroxide is present in a concentration of 2.5% to 10%, for example, about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% (weight / volume). In an embodiment, the benzoyl peroxide is present in a concentration of less than 2.5% but greater than about 0.1%, 0.5%, 1%, 1.5%, or 2% (weight / volume). In an embodiment, salicylic acid is present in a concentration of 0.5% to 2%, for example, about 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9% or 2% (weight / volume). In an embodiment, salicylic acid is present in a concentration of less than 0.5% but greater than about 0.1% (weight / volume). In an embodiment, sulfur is present in a concentration of 3% to 10%, for example, about 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10% (weight / volume). In embodiments, sulfur is present at a concentration of less than 3% but greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight / volume). In embodiments, resorcinol is present at a concentration of 2% and sulfur is present at a concentration of 3%-8% (e.g., about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5% or 8%) (weight / volume). In embodiments, resorcinol monoacetate is present at a concentration of 3% and sulfur is present at a concentration of 3%-8% (e.g., about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5% or 8%) (weight / volume).In embodiments, resorcinol is present at a concentration less than 2% but greater than about 0.1%, 0.5%, 1%, 1.5% (weight / volume), hi embodiments, resorcinol monoacetate is present at a concentration less than 3% but greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight / volume).

[0172] In embodiments, the compositions provided herein comprise a moisturizer.

[0173] How to Treat Acne In an aspect, provided herein is a method of preventing or treating acne in a subject in need of such treatment, the method comprising administering an effective amount of a composition or combination provided herein. In an embodiment, an effective amount of a composition comprising, consisting essentially of, or consisting of at least one P. acnes bacteriophage, at least one anti-acne compound, and a pharma- ceutically acceptable carrier is administered to the subject. In an embodiment, an effective amount of a composition comprising at least one P. acnes bacteriophage, at least one anti-acne compound, and a pharma- ceutically acceptable carrier, the composition not including a probiotic bacterium, is administered to the subject. In an embodiment, an effective amount of a composition comprising a P. acnes bacteriophage and an enzyme is administered to the subject.

[0174] In embodiments, an effective amount of a composition comprising a bacteriophage described herein, including this embodiment, is administered to a subject. In embodiments, the bacteriophage is a wild-type bacteriophage.

[0175] In an embodiment, the bacteriophage is administered topically. In an embodiment, the bacteriophage is in a composition (e.g., a pharmaceutical or cosmetic composition) that further comprises a pharma- ceutically or cosmetically acceptable carrier.

[0176] In an embodiment, the method further comprises administering probiotic bacteria to the subject.

[0177] In an aspect, a method of treating acne in a subject in need thereof is provided. The method comprises administering to the subject an effective amount of probiotic P. acnes bacteria. In an embodiment, the method further comprises administering to the subject a bacteriophage.

[0178] In an embodiment, the P. acnes has a 16S rDNA sequence comprising T992C, T838C, C1322T and / or C986T mutations compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO: 2. In an embodiment, the P. acnes comprises a 16S rDNA sequence with T838C and C1322T mutations compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO: 2. In an embodiment, the P. acnes is a ProI strain. In an embodiment, the P. acnes comprises a 16S rDNA sequence with C986T and T992C mutations compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO: 2. In an embodiment, the P. acnes is a ProII strain.

[0179] In an embodiment, the P. acnes bacteria (a) comprises a 16S rDNA sequence having a T992C mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (b) comprises a 16S rDNA sequence having a T838C mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (c) comprises a 16S rDNA sequence having a C1322T mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (d) comprises a 16S rDNA sequence having a C986T mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (e) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO:3; (g) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO:4. (h) does not contain linear plasmids; (i) does not contain plasmids containing virulence factors; and / or (j) does not contain plasmids encoding extrachromosomal lipases and / or adhesive virulence factors.

[0180] In an embodiment, the method further comprises administering to the subject at least one additional probiotic bacteria.

[0181] In an embodiment, the at least one additional bacterium comprises, consists essentially of, or consists of probiotic bacteria. In an embodiment, the at least one bacterium comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains and / or species, less than about 10, 9, 8, 7, 6, 5, 4, 3, or 2 bacterial strains and / or species, or 1-10, 2-10, 3-10, 4-10, 5-10, 1-5, 2-5, 3-5, or 4-5 bacterial strains and / or species. In an embodiment, the at least one bacterium comprises a plurality of bacterial strains and / or species, for example at least about 2, 3, 4, 5, 6, 7, 8, 9, 10 bacterial strains and / or species. In an embodiment, the at least one bacterium comprises Propionibacterium spp., Staphylococcus spp., and / or Corynebacterium spp. bacteria. In an embodiment, the at least one bacterium comprises a bacterium in the class Betaproteobacteria. In an embodiment, the at least one bacterium comprises an isolated Propionibacterium granulosum bacterium, an isolated Propionibacterium avidum bacterium, an isolated Staphylococcus epidermidis bacterium, an isolated Staphylococcus aureus bacterium, and / or an isolated Corynebacterium jacqueium bacterium. In an embodiment, the at least one bacterium comprises one, two, three, four or five of an isolated Propionibacterium granulosum bacterium, an isolated Propionibacterium avidum bacterium, an isolated Staphylococcus epidermidis bacterium, an isolated Staphylococcus aureus bacterium, and / or an isolated Corynebacterium jacqueium bacterium.

[0182] In an embodiment, the subject has been administered a bacteriophage described herein, including this embodiment.

[0183] In an embodiment, the subject is administered an antibiotic that kills P. acnes, hi an embodiment, the antibiotic is clindamycin, doxycycline, erythromycin, or tetracycline, or a derivative of clindamycin, doxycycline, erythromycin, or tetracycline.

[0184] In embodiments, the antibiotic is clindamycin, doxycycline, erythromycin, or tetracycline, or a derivative of clindamycin, doxycycline, erythromycin, or tetracycline.

[0185] In an embodiment, the method further comprises administering to the subject an enzyme, such as a biofilm degrading enzyme or an anti-aging enzyme. Non-limiting examples of biofilm degrading enzymes include DNases (e.g., DNase I), restriction endonucleases, proteases (e.g., papain, bromelain, trypsin, proteinase K, subtilisin or serratiopeptidase), glycosidases (e.g., dispersin, alginate lyase, amylase or cellulase). Non-limiting examples of anti-aging enzymes include superoxide dismutase and peroxidase.

[0186] In embodiments, the method further comprises administering a topical retinoid, an antibiotic, and / or an alpha hydroxy acid. In embodiments, the method further comprises administering a topical retinoid. In embodiments, the method further comprises administering an antibiotic. In embodiments, the method further comprises administering an alpha hydroxy acid. In embodiments, the method further comprises administering benzoyl peroxide, salicylic acid, sulfur, resorcinol, and / or resorcinol monoacetate to the subject. In embodiments, the method further comprises administering benzoyl peroxide. In embodiments, the method further comprises administering salicylic acid. In embodiments, the method further comprises administering sulfur. In embodiments, the method further comprises administering resorcinol and / or sulfur. In embodiments, the method further comprises administering resorcinol and / or resorcinol monoacetate.

[0187] In embodiments, the method further comprises administering a enhancing peptide or enzyme, which in embodiments has one or more or any combination of the following properties: biofilm degradation, improved skin penetration, antibacterial, reduced inflammation (e.g., of the skin), reduced irritation (e.g., of the skin), reduced redness (e.g., of the skin), skin tightening, wrinkle removal, fine line removal, or other improvement in appearance (e.g., of the skin).

[0188] In embodiments, at least about 2, 3, 4, 5, 6, 7, 8, 9, or 10 P. acnes bacteriophages are administered to the subject. In embodiments, the P. acnes bacteriophage comprises more than one type of P. acnes bacteriophage.

[0189] Exemplary Methods and Compositions for Treating Acne In an aspect, a composition comprising a bacteriophage is provided herein. In an embodiment, the bacteriophage is present in a composition, such as a therapeutic or cosmetic composition. In an embodiment, the composition further comprises a strain of probiotic bacteria. In an embodiment, the composition further comprises an enzyme that degrades bacterial biofilms (e.g., components thereof) in or on human skin pores. In an embodiment, the enzyme enhances the penetration of the bacteriophage and / or the probiotic bacteria. In an embodiment, the bacteriophage ("phage") destroys acne-causing (i.e., pathogenic) strains of P. acnes with high specificity and efficacy, without killing beneficial skin bacteria. In an embodiment, the biofilm-degrading enzyme dissolves the biofilm to increase the susceptibility of pathogens (e.g., by reducing pathogen adhesion to host cells and / or by increasing the access of the bacteriophage to pathogenic cells). In embodiments, the probiotic bacteria are immune to the bacteriophage (e.g., the bacteria lack a cellular receptor to which the bacteriophage specifically binds). In embodiments, the probiotic bacteria occupy the niche left by a killed pathogenic strain of P. acnes. In embodiments, the probiotic bacteria reduce or prevent recolonization or growth on the subject's skin (such as pores) by surviving longer than the pathogenic bacteria.

[0190] In an aspect, a composition is provided for the therapeutic treatment of the skin disease acne. In an embodiment, the composition comprises a lytic P. acnes bacteriophage, optionally a probiotic bacterium originating from healthy skin, and / or optionally a biofilm degrading enzyme in the composition as an adjuvant to enhance the penetration of the active components.

[0191] In an embodiment, the lytic P. acnes bacteriophage infects pathogenic P. acnes in skin comedones. In an embodiment, the bacteriophage replicates within and lyses P. acnes. In an embodiment, upon lysis, P. acnes releases new virions. In an embodiment, the enzyme unclogs blocked comedones, lyses the P. acnes biofilm, and increases the access of virions to P. acnes. In an embodiment, the exponential growth of the lytic P. acnes phage rapidly kills P. acnes with high specificity without interfering with the growth of beneficial commensal skin bacteria. In an embodiment, the niche vacated by P. acnes is then filled by probiotic bacteria. In an embodiment, the bacteria originate from healthy skin and expand to occupy the niche, thereby preventing any surviving P. acnes bacteria from growing back. In an embodiment, this strategy helps balance the skin microbiome in a subject, rebalancing the microbiome towards a healthy skin bacterial population. In an embodiment, biofilm degrading enzymes are present in the formulation as adjuvants to help unclog blocked comedones and increase access of phages and probiotic bacteria to the pores.

[0192] In an embodiment, a combination is provided that includes a bacteriophage, a probiotic bacterium, and (optionally) an enzyme that enhances the penetration of the bacteriophage. In an embodiment, the pathogenic bacteria are killed and the probiotic bacteria replace the pathogenic bacteria. In an embodiment, a "kill and replace" approach is used to treat acne. In an embodiment, a biologic that selectively kills acne-causing pathogenic bacteria is administered to the subject. In an embodiment, probiotic bacteria originating from healthy skin are applied to occupy the niche of the killed pathogenic bacteria. In an embodiment, this approach avoids the problem of intractable drug resistance associated with antibiotics. In an embodiment, the presence of actively dividing probiotic bacteria prevents recurrence by preventing any pathogenic bacteria from growing back. In an embodiment, dysbiosis on the skin of the subject is treated. In an embodiment, the microbiome associated with acne is rebalanced to a healthy microbiome.

[0193] In an embodiment, the bacteriophage is a naturally occurring P. acnes bacteriophage.

[0194] Non-limiting examples of enzymes that may be co-administered with a bacteriophage include BL00275 from Bacillus licheniformis; deoxyribonuclease I; restriction endonucleases, deoxyribonucleases (e.g., from Staphylococcus aureus thermonuclease, B. licheniformis NucB, deoxyribonuclease 1L2); glycoside hydrolases (e.g., dispersin B, alginate lyase, amylase, cellulase, glycanase); and proteases (e.g., subtilisin, proteinase K, trypsin, serratiopeptidase).

[0195] Non-limiting examples of probiotic bacteria that may be administered or present in the system or composition include one or more or any combination of the following bacterial species: Propionibacterium acnes, Propionibacterium granulosum, Propionibacterium avidum, Staphylococcus epidermidis, Staphylococcus aureus, and Corynebacterium jacqueum. In embodiments, the probiotic bacterial strain is selected based on (a) its ability to colonize the skin without eliciting a deleterious immune response, characterized by low lipase activity and low adhesion to human keratinocytes; and (b) its ability to occupy a niche similar to Propionibacterium acnes.

[0196] In embodiments, biofilm degrading enzymes are present in the formulation to act as adjuvants to enhance the effectiveness of active ingredients (such as bacteriophages), hi embodiments, the enzymes have the ability to degrade P. acnes biofilms in vitro.

[0197] Embodiment In order to facilitate a more complete understanding of the present invention, the following embodiments and examples are provided. The following embodiments and examples set forth exemplary modes of making and carrying out the present invention. However, the scope of the present invention is not limited to the specific embodiments disclosed in these embodiments and examples, which are for illustrative purposes only, since alternative methods may be utilized to obtain similar results.

[0198] Embodiments include the following embodiments P1 to P56.

[0199] Embodiment P1. A composition consisting essentially of at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound and a pharma- ceutically acceptable carrier.

[0200] Embodiment P2. A composition comprising at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound and a pharma- ceutically acceptable carrier, and does not include probiotic bacteria.

[0201] Embodiment P3. The composition of embodiment P2, wherein the composition further comprises a P. acnes biofilm degrading enzyme.

[0202] Embodiment P4. The composition of any one of Embodiments P1-P3, wherein at least one anti-acne compound is benzoyl peroxide.

[0203] Embodiment P5. The composition of embodiment P4 wherein the benzoyl peroxide is present in a concentration of 2.5% to 10% (weight / volume).

[0204] Embodiment P6. The composition of embodiment P4 wherein benzoyl peroxide is present at a concentration less than 2.5% but greater than about 0.1%, 0.5%, 1%, 1.5% or 2% (weight / volume).

[0205] Embodiment P7. The composition of any one of embodiments P1-P3, wherein at least one anti-acne compound is salicylic acid.

[0206] Embodiment P8. The composition of embodiment P7 wherein the salicylic acid is present in a concentration of 0.5% to 2% (weight / volume).

[0207] Embodiment P9. The composition of embodiment P7 wherein salicylic acid is present at a concentration less than 0.5% but greater than about 0.1% (weight / volume).

[0208] Embodiment P10. The composition of any one of Embodiments P1-P3, wherein at least one anti-acne compound is sulfur.

[0209] Embodiment P11. The composition of embodiment P10 wherein the sulfur is present in a concentration of 3% to 10% (weight / volume).

[0210] Embodiment P12. The composition of embodiment P10 wherein sulfur is present at a concentration less than 3% but greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight / volume).

[0211] Embodiment P13. The composition of any one of Embodiments P1-P3, wherein at least one anti-acne compound is resorcinol and sulfur.

[0212] Embodiment P14. The composition of embodiment P13 wherein the resorcinol is present in a concentration of 2% and the sulfur is present in a concentration of 3% to 8% (weight / volume).

[0213] Embodiment P15. The composition of any one of Embodiments P1-P3 wherein the at least one anti-acne compound comprises resorcinol monoacetate and sulfur.

[0214] Embodiment P16. The composition of embodiment P15 wherein the resorcinol monoacetate is present in a concentration of 3% and the sulfur is present in a concentration of 3% to 8% (weight / volume).

[0215] Embodiment P17. The composition of any one of Embodiments P1-P3 wherein the anti-acne compound is an antibiotic, a retinoid, or an alpha hydroxy acid.

[0216] Embodiment P18. A composition comprising a Propionibacterium acnes bacteriophage and an enzyme.

[0217] Embodiment P19. The composition of any one of embodiments P1 to P18, wherein the P. acnes bacteriophage is a lytic P. acnes bacteriophage.

[0218] Embodiment P20. The composition of any one of embodiments P1-P19, wherein the P. acnes bacteriophage comprises a linear double-stranded DNA genome.

[0219] Embodiment P21. The composition of any one of embodiments P1 to P20, wherein the P. acnes bacteriophage is within the bacteriophage Siphoviridae family.

[0220] Embodiment P22. The composition of any one of embodiments P1 to P21, wherein the genome of the P. acnes bacteriophage comprises a nucleotide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleotide sequence of SEQ ID NO:1.

[0221] Embodiment P23. The composition of any one of embodiments P18 to P21, wherein the enzyme is a P. acnes biofilm-degrading enzyme.

[0222] Embodiment P24. The composition of any one of embodiments P3 or P18-P23, wherein the enzyme is a glycosidase, protease, deoxyribonuclease or restriction endonuclease.

[0223] Embodiment P25. The composition of any one of embodiments P3 or P18-P24, wherein the enzyme is a glycosidase.

[0224] Embodiment P26. The composition of embodiment P25, wherein the glycosidase is a glycoside hydrolase.

[0225] Embodiment P27. The composition of embodiment P26, wherein the enzyme catalyzes the hydrolysis of linear polymers of N-acetyl-D-glucosamine.

[0226] Embodiment P28. The composition of embodiment P27, wherein the enzyme is β-hexosaminidase.

[0227] Embodiment P29. The composition of embodiment P28, wherein the enzyme hydrolyzes the β-1,6-glycosidic bonds of the acetylglucosamine polymer.

[0228] Embodiment P30. The composition of any one of embodiments P3 or P18-P24, wherein the enzyme is deoxyribonuclease I, a restriction endonuclease, papain, bromelain, trypsin, proteinase K, subtilisin, serratiopeptidase, dispersin, alginate lyase, amylase or cellulase.

[0229] Embodiment P31. The composition of any one of embodiments P3 or P18-P24, wherein the enzyme is dispersin B.

[0230] Embodiment P32. The composition of any one of embodiments P3 or P18-P24, wherein the enzyme is a protease, wherein the protease is proteinase K or subtilisin.

[0231] Embodiment P33. The composition of any one of embodiments P18 to P22, wherein the enzyme is an anti-aging enzyme.

[0232] Embodiment P34. The composition of embodiment P33, wherein the anti-aging enzyme is superoxide dismutase or peroxidase.

[0233] Embodiment P35. The composition of any one of embodiments P18 to P34 further comprising probiotic bacteria.

[0234] Embodiment P36. The composition of embodiment P35, wherein the probiotic bacteria are probiotic P. spp., Staphylococcus spp. and / or Corynebacterium spp. bacteria.

[0235] Embodiment P37. The composition of embodiment P35, wherein the probiotic bacteria is a bacterium within the class Betaproteobacteria.

[0236] Embodiment P38. The composition of embodiment P36, wherein the probiotic bacteria is probiotic P. acnes.

[0237] Embodiment P39. The P. acnes bacteria (a) comprising a 16S DNA sequence having a T992C mutation compared to the KPA171202 reference strain 16S DNA sequence set forth as SEQ ID NO:2; (b) comprising a 16S DNA sequence having a T838C mutation relative to the KPA171202 reference strain 16S DNA sequence set forth as SEQ ID NO:2; (c) comprising a 16S DNA sequence having a C1322T mutation compared to the KPA171202 reference strain 16S DNA sequence set forth as SEQ ID NO:2; (d) comprising a 16S DNA sequence having a C986T mutation compared to the KPA171202 reference strain 16S DNA sequence set forth as SEQ ID NO:2; (e) comprises a 16S DNA sequence identical to the sequence of SEQ ID NO:3; (f) comprises a 16S DNA sequence identical to the sequence of SEQ ID NO:4; (g) does not contain linear plasmid; (h) does not contain a plasmid containing a virulence factor; and / or (i) does not contain plasmids encoding extrachromosomal lipases and / or adhesion virulence factors; The composition of embodiment P38.

[0238] Embodiment P40. The P. acnes bacteria (a) produces less than about 20% of the level of lipase produced by pathogenic P. acnes strains when grown in planktonic culture; (b) produces less than about 10% of the level of lipase produced by pathogenic P. acnes strains when grown in adherent culture; (c) adheres to epithelial cells at least 50% less than a pathogenic P. acnes strain; and / or (d) less inflammatory than pathogenic P. acnes strains; The composition of embodiment P38.

[0239] Embodiment P41. The composition of any one of embodiments P35 to P40, further comprising at least one additional probiotic bacteria.

[0240] Embodiment P42. The composition of embodiment P41, wherein the at least one additional probiotic bacterium comprises Propionibacterium granulosum and / or Propionibacterium avidum.

[0241] Embodiment P43. The pathogenic P. acnes strain is (a) comprising a 16S DNA sequence having a G1058C mutation compared to the KPA171202 reference strain 16S DNA sequence set forth as SEQ ID NO:2; (b) comprising a 16S DNA sequence having a G1058C and A1201C mutation compared to the KPA171202 reference strain 16S DNA sequence set forth as SEQ ID NO:2; (c) comprising a 16S DNA sequence having a G529A mutation compared to the KPA171202 reference strain 16S DNA sequence set forth as SEQ ID NO:2; (d) comprising a 16S DNA sequence having a G1004A and a T1007C mutation relative to the KPA171202 reference strain 16S DNA sequence set forth as SEQ ID NO:2; (e) comprising a 16S DNA sequence having a G1268A mutation compared to the KPA171202 reference strain 16S DNA sequence set forth as SEQ ID NO:2; (f) comprising a 16S DNA sequence having a T554C and a G1058C mutation relative to the KPA171202 reference strain 16S DNA sequence set forth as SEQ ID NO:2; (g) comprises a 16S DNA sequence identical to the sequence of SEQ ID NO:5; (h) comprises a 16S DNA sequence identical to the sequence of SEQ ID NO:6; (i) comprises a 16S DNA sequence identical to the sequence of SEQ ID NO:7; (j) comprises a 16S DNA sequence identical to the sequence of SEQ ID NO:8; (k) comprises a 16S DNA sequence identical to the sequence of SEQ ID NO: 9; and / or (l) comprises a 16S DNA sequence identical to the sequence of SEQ ID NO: 10; The composition of embodiment P40.

[0242] Embodiment P44. The composition of any one of embodiments P18 to P43, further comprising at least one additional P. acnes bacteriophage.

[0243] Embodiment P45. The composition of any one of embodiments P1-P44 comprising a pharma- ceutically acceptable carrier.

[0244] Embodiment P46. The composition of embodiment P45, wherein the pharma- ceutically acceptable carrier comprises an emulsion.

[0245] Embodiment P47. The composition of embodiment P46, wherein the emulsion is an oil-in-water emulsion or a water-in-oil emulsion.

[0246] Embodiment P48. The composition of any one of embodiments P1 to P47 which is in the form of a cream, lotion, suspension or aqueous solution.

[0247] Embodiment P49. A combination consisting essentially of at least one Propionibacterium acnes bacteriophage and at least one anti-acne compound, wherein each of the at least one Propionibacterium acnes bacteriophage and the at least one anti-acne compound is in a composition further comprising a pharma- ceutically acceptable carrier.

[0248] Embodiment P50. The combination of embodiment P49, wherein the at least one P. acnes bacteriophage and the at least one anti-acne compound are in separate compositions.

[0249] Embodiment P51. The combination of embodiment P50, wherein the at least one P. acnes bacteriophage and the at least one anti-acne compound are in separate containers.

[0250] Embodiment P52. A combination comprising a Propionibacterium acnes bacteriophage and an enzyme.

[0251] Embodiment P53. The combination of embodiment P52, wherein the P. acnes bacteriophage and the enzyme are in separate compositions.

[0252] Embodiment P54. The combination of embodiment P53, wherein the P. acnes bacteriophage and the enzyme are in separate containers.

[0253] Embodiment P55. A method of treating acne in a subject in need thereof comprising administering to the subject an effective amount of a composition of any one of embodiments P1-P46 or a combination of any one of embodiments P49-P54.

[0254] Embodiment P56. The method of embodiment P55, wherein the composition is administered topically.

[0255] Additional embodiments include the following embodiments 1-55.

[0256] Embodiment 1. A composition comprising at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound, and a pharma- ceutically acceptable carrier.

[0257] Embodiment 2. The composition of embodiment 1, wherein the composition does not comprise probiotic bacteria.

[0258] Embodiment 3. The composition of embodiment 1 or 2, wherein the composition further comprises a P. acnes biofilm-degrading enzyme.

[0259] Embodiment 4. The composition of any one of embodiments 1-3, wherein at least one anti-acne compound is salicylic acid.

[0260] Embodiment 5. The composition of embodiment 4, wherein the salicylic acid is present in a concentration of 0.5% to 2% (weight / volume).

[0261] Embodiment 6. The composition of embodiment 5, wherein salicylic acid is present at a concentration of less than 0.5% but greater than about 0.1% (weight / volume).

[0262] Embodiment 7. The composition of any one of embodiments 1-3, wherein at least one anti-acne compound is sulfur.

[0263] Embodiment 8. The composition of embodiment 7, wherein the sulfur is present in a concentration of 3% to 10% (weight / volume).

[0264] Embodiment 9. The composition of embodiment 7, wherein sulfur is present at a concentration of less than 3% but greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight / volume).

[0265] Embodiment 10. The composition of any one of embodiments 1-3, wherein at least one anti-acne compound is resorcinol and sulfur.

[0266] Embodiment 11. The composition of embodiment 10, wherein the resorcinol is present in a concentration of 2% and the sulfur is present in a concentration of 3% to 8% (weight / volume).

[0267] Embodiment 12. The composition of any one of embodiments 1-3, wherein the at least one anti-acne compound comprises resorcinol monoacetate and sulfur.

[0268] Embodiment 13. The composition of embodiment 12, wherein the resorcinol monoacetate is present in a concentration of 3% and the sulfur is present in a concentration of 3% to 8% (weight / volume).

[0269] Embodiment 14 The composition of any one of embodiments 1-3, wherein the anti-acne compound is an antibiotic, a retinoid, or an alpha hydroxy acid.

[0270] Embodiment 15. The composition of any one of embodiments 1 to 14, wherein the Propionibacterium acnes bacteriophage is a naturally occurring Propionibacterium acnes bacteriophage.

[0271] Embodiment 16. The composition of any one of embodiments 1 to 15, wherein the P. acnes bacteriophage is a lytic P. acnes bacteriophage.

[0272] Embodiment 17. The composition of any one of embodiments 1-16, wherein the P. acnes bacteriophage comprises a linear double-stranded DNA genome.

[0273] Embodiment 18. The composition of any one of embodiments 1-17, wherein the P. acnes bacteriophage is within the bacteriophage Siphoviridae family.

[0274] Embodiment 19. The composition of any one of embodiments 1 to 19, wherein the genome of the P. acnes bacteriophage comprises a nucleotide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleotide sequence of SEQ ID NO:1.

[0275] Embodiment 20. The composition of any one of embodiments 3 to 19, wherein the enzyme is a P. acnes biofilm-degrading enzyme.

[0276] Embodiment 21. The composition of any one of embodiments 3 to 20, wherein the enzyme is a glycosidase, protease, deoxyribonuclease or restriction endonuclease.

[0277] Embodiment 22 The composition of any one of embodiments 3 to 21, wherein the enzyme is a glycosidase.

[0278] Embodiment 23 The composition of embodiment 22, wherein the glycosidase is a glycoside hydrolase.

[0279] Embodiment 24 The composition of embodiment 23, wherein the enzyme catalyzes the hydrolysis of linear polymers of N-acetyl-D-glucosamine.

[0280] Embodiment 25 The composition of embodiment 24, wherein the enzyme is β-hexosaminidase.

[0281] Embodiment 26 The composition of embodiment 25, wherein the enzyme hydrolyzes the β-1,6-glycosidic bonds of the acetylglucosamine polymer.

[0282] Embodiment 27. The composition of any one of embodiments 3 to 20, wherein the enzyme is deoxyribonuclease I, restriction endonuclease, papain, bromelain, trypsin, proteinase K, subtilisin, serratiopeptidase, dispersin, alginate lyase, amylase or cellulase.

[0283] Embodiment 28. The composition of any one of embodiments 3 to 20, wherein the enzyme is dispersin B.

[0284] Embodiment 29. The composition of any one of embodiments 3 to 20, wherein the enzyme is a protease, and the protease is proteinase K or subtilisin.

[0285] Embodiment 30. The composition of any one of embodiments 1 to 29, further comprising an anti-aging enzyme.

[0286] Embodiment 31 The composition of embodiment 30, wherein the anti-aging enzyme is superoxide dismutase or peroxidase.

[0287] Embodiment 32. The composition of any one of embodiments 1 to 31, further comprising probiotic bacteria.

[0288] Embodiment 33. The composition of embodiment 32, wherein the probiotic bacteria are probiotic P. spp., Staphylococcus spp. and / or Corynebacterium spp. bacteria.

[0289] Embodiment 34 The composition of embodiment 32, wherein the probiotic bacteria is a bacterium within the class Betaproteobacteria.

[0290] Embodiment 35 The composition of embodiment 33, wherein the probiotic bacteria is probiotic P. acnes.

[0291] 36. The P. acnes bacteria is (a) comprising a 16S rDNA sequence having a T992C mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (b) comprising a 16S rDNA sequence having a T838C mutation relative to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (c) comprising a 16S rDNA sequence having a C1322T mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (d) comprising a 16S rDNA sequence having a C986T mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (e) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO:3; (f) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO:4; (g) does not contain linear plasmid; (h) does not contain a plasmid containing a virulence factor; and / or (i) does not contain plasmids encoding extrachromosomal lipases and / or adhesion virulence factors; The composition of embodiment 35.

[0292] Embodiment 37. The P. acnes bacteria is (a) produces less than about 20% of the level of lipase produced by pathogenic P. acnes strains when grown in planktonic culture; (b) produces less than about 10% of the level of lipase produced by pathogenic P. acnes strains when grown in adherent culture; (c) adheres to epithelial cells at least 50% less than a pathogenic P. acnes strain; and / or (d) less inflammatory than pathogenic P. acnes strains; The composition of embodiment 35 or 36.

[0293] Embodiment 38. The composition of any one of embodiments 32 to 37, further comprising at least one additional probiotic bacteria.

[0294] Embodiment 39. The composition of embodiment 38, wherein the at least one additional probiotic bacterium comprises Propionibacterium granulosum and / or Propionibacterium avidum.

[0295] Embodiment 40. The pathogenic P. acnes strain is (a) comprising a 16S rDNA sequence having a G1058C mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (b) comprising a 16S rDNA sequence having G1058C and A1201C mutations relative to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (c) comprising a 16S rDNA sequence having a G529A mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (d) comprising a 16S rDNA sequence having G1004A and T1007C mutations relative to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (e) comprising a 16S rDNA sequence having a G1268A mutation compared to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (f) comprising a 16S rDNA sequence having T554C and G1058C mutations relative to the KPA171202 reference strain 16S rDNA sequence set forth as SEQ ID NO:2; (g) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO:5; (h) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO:6; (i) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO:7; (j) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO:8; (k) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO: 9; and / or (l) comprises a 16S rDNA sequence identical to the sequence of SEQ ID NO: 10; The composition of embodiment 37.

[0296] Embodiment 41 The composition of any one of embodiments 1 to 40, further comprising at least one additional P. acnes bacteriophage.

[0297] Embodiment 42. The composition of any one of embodiments 1 to 41, wherein the pharma- ceutically acceptable carrier comprises an emulsion.

[0298] Embodiment 43. The composition of embodiment 42, wherein the emulsion is an oil-in-water emulsion or a water-in-oil emulsion.

[0299] Embodiment 44. The composition of any one of embodiments 1 to 44, which is in the form of a cream, lotion, suspension or aqueous solution.

[0300] Embodiment 45. A combination comprising at least one Propionibacterium acnes bacteriophage and at least one anti-acne compound, wherein each of the at least one Propionibacterium acnes bacteriophage and the at least one anti-acne compound is in a composition further comprising a pharma- ceutically acceptable carrier.

[0301] Embodiment 46 The combination of embodiment 45, wherein the at least one P. acnes bacteriophage and the at least one anti-acne compound are in separate compositions.

[0302] Embodiment 47. The combination of embodiment 46, wherein at least one anti-acne compound is benzoyl peroxide.

[0303] Embodiment 48. The combination of embodiment 47, wherein the benzoyl peroxide is present in a concentration of 2.5% to 10% (weight / volume).

[0304] Embodiment 49. The combination of embodiment 47, wherein benzoyl peroxide is present at a concentration of less than 2.5% but greater than about 0.1%, 0.5%, 1%, 1.5% or 2% (weight / volume).

[0305] Embodiment 50. A method of treating acne in a subject in need thereof, comprising administering to the subject an effective amount of the composition of any one of embodiments 1-44.

[0306] Embodiment 51 The method of embodiment 50, wherein the composition is administered topically.

[0307] Embodiment 52. A method of treating acne in a subject in need thereof, comprising administering to the subject an effective amount of the combination of any one of embodiments 45-49.

[0308] Embodiment 53. A composition comprising a Propionibacterium acnes bacteriophage and an enzyme.

[0309] Embodiment 54. A combination comprising a Propionibacterium acnes bacteriophage and an enzyme.

[0310] Embodiment 55. A composition consisting essentially of at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound, and a pharma- ceutically acceptable carrier. EXAMPLES

[0311] The following examples illustrate certain embodiments of the invention and are not intended to limit the scope of the invention.

[0312] The embodiments of the present specification are further described by the following examples and detailed protocols.However, the examples are only intended to illustrate the embodiments and should not be interpreted as limiting the scope of the present specification.The contents of all references and published patents and patent applications cited throughout this application are hereby incorporated by reference.

[0313] Example 1: P. acnes bacteriophage PHIT-101 selectively and efficiently kills P. acnes P. acnes bacteriophages have been shown to be genetically highly similar and to exhibit broad range against multiple strains of P. acnes. The lead bacteriophage (PHIT-101) was used for the experiments. PHIT-101 is the only lytic phage that killed all strain types of P. acnes tested (data not shown). PHIT-101 has the sequence of SEQ ID NO:1. To demonstrate the efficacy and specificity of this phage, a plate assay was performed as follows: P. acnes KPA171202 and P. granulosum (a closely related but benign skin bacterium) were plated on separate BHI agar plates. A sterile cotton pad was placed on each plate. The sterile cotton pad was coated with minocycline, an antibiotic commonly used to treat acne, or 2×10 7 Plates were immersed in phage solutions with titers of 0.01 pfu / mL. After 72 h of anaerobically incubation at 37°C, the minocycline pads indiscriminately killed bacteria, exhibiting a kill zone against both acne-causing P. acnes and commensal P. granulosum (Figure 1). In contrast, the PHIT-101 pads killed only P. acnes and did not impede the growth of beneficial P. granulosum.

[0314] Further evidence of the ability of PHIT-101 to selectively kill bacteria was obtained in a synthetic skin microbiome assay. A synthetic skin microbiome was devised containing P. acnes, P. granulosum and P. avidum, three skin bacteria that make up 60-80% of the bacterial flora in skin pores [Science (2009) 324:1190-1192]. This synthetic skin microbiome was grown anaerobically in the presence or absence of PHIT-101 (final concentration 5 × 10 5pfu / mL). After 48 h incubation at 37 °C, cells were pelleted and washed, and the relative proportions of the three bacterial species were determined using 16S amplicon next-generation sequencing (NGS) on an Illumina MiSeq. The results in Figure 2 show that PHIT-101 can almost completely kill P. acnes and has no negative effect on the growth of the symbiotic P. granulosum and P. avidum.

[0315] Screening biofilm-degrading enzymes (BDEs) for disrupting P. acnes biofilms Several recent reports (Exp Dermatol (2014) 23:687; Br J Dermatol (2015) 172:13) have established that P. acnes produces significant amounts of biofilm in skin pores, which impedes antibiotic penetration and leads to poor treatment outcomes. To substantiate this, biofilm production of several strains of P. acnes was quantified. Figure 3 shows that adherent cultures of multiple strains isolated from a single subject's microbiota produce significantly different levels of biofilm under similar conditions. Thus, previous proofs of concept using planktonic cells did not reflect the true conditions in which P. acnes grows on the skin.

[0316] Without being bound by any scientific theory, we hypothesized that biofilms may be a significant barrier to phage killing of sessile P. acnes cells. This hypothesis was confirmed in cell viability assays (Figure 5) which showed that PHIT-101 was only able to kill approximately 50% of P. acnes cells encased in biofilms, unlike planktonic P. acnes (99% killing, Figure 2). To determine whether biofilm degradation would improve phage killing, several enzymes were screened to find BDEs specific to P. acnes. The screen included three classes of enzymes that could degrade types of materials likely to be found in biofilms: DNA, polysaccharides, and proteins. Figure 4 shows that in the screen, deoxyribonucleases had moderate activity, and the best biofilm degradation rates were found with proteases and dispersin, a glycoside hydrolase from Aggregatibacter actinomycetemcomitans.

[0317] In selecting a BDE to pair with the phage, dispersin was chosen for two reasons. First, being a glycoside hydrolase, dispersin could not attack the protein membrane of the phage itself, thereby avoiding possible degradation of the phage. Second, P. acnes forms a robust biofilm with Staphylococcus aureus [Anaerobe (2016) 40:63-67], and dispersin is active against biofilms from both organisms. We determined whether the addition of dispersin would increase the efficiency of phage killing in sessile P. acnes. Figure 5 shows that killing of PHIT-101 was enhanced in the presence of dispersin, restoring approximately 99% killing efficiency of the phage.

[0318] [Example 2] Probiotic bacteria Genotypic characterization of probiotic strains Strains of P. acnes were characterized based on point mutations in the 16S rDNA sequence leading to a phylogenetic classification into pathogenic and probiotic strain types, and the absence of linear plasmids found in pathogenic strains carrying virulence factors. The complete 16S rDNA sequence of each P. acnes strain was amplified and Sanger sequenced using 16S-specific primers. Probiotic strains were confirmed to have ProI and ProII ribosequences (RS). ProI strains have T838C and C1322T mutations compared to the 16S rDNA sequence of the KPA171202 reference strain (NIH accession number NC_006085.1). ProII strains have C986T and T992C mutations compared to the KPA171202 sequence. In addition, the presence or absence of linear plasmids in each strain was determined using specific primer pairs. The probiotic strain was confirmed to lack this plasmid, which carries the extrachromosomal lipase as well as the Tad (adherence) virulence factor.

[0319] In an embodiment, the probiotic strain is characterized primarily by its 16S sequence, e.g., SEQ ID NO: 3 and SEQ ID NO: 4. In an embodiment, the probiotic strain can be genotypically identified by the lack of extrachromosomal lipases and plasmids carrying virulence factors such as the Tad locus.

[0320] A cohort of probiotic strains was further characterized for their immunogenic potential. Lead probiotic candidates were based on two factors: low lipase production and reduced adhesion to epithelial cells. Phenotypic confirmation of these features was important in selecting probiotic lead candidates.

[0321] Testing the immunogenic potential of probiotic P. acnes strains: Lipase activity Lipase plays a key role in acne pathogenesis by hydrolyzing sebaceous triglycerides and releasing irritating free fatty acids into pilosebaceous follicles. Lipase is a strong chemotactic and proinflammatory antigen. Therefore, lipase is of great interest as a pharmacological target for anti-acne drugs. In an embodiment, the overall strategy is to replace pathogenic P. acnes, which secretes high levels of lipase, with low-secreting probiotic P. acnes. To quantify the lipase expression phenotype for each strain in our panel, lipase production of probiotic P. acnes strains was compared to pathogenic P. acnes strains using a fluorescent lipase activity assay.

[0322] One of the most interesting findings was that each strain secreted different amounts of lipase when grown in planktonic versus attached culture. This was previously reported for P. acnes strains [Res Microbiol, (2007) 158:386-392]. Furthermore, the data showed that there was no significant difference between the lipase production of the pathogenic and probiotic strains when these strains were grown in liquid culture. However, interesting changes were observed when these strains were grown under biofilm conditions. Although the variability in production between strains could still be observed, some probiotic strains had significantly less lipase activity than the pathogenic strains (Figure 11). Interestingly, not all strains in the probiotic cohort had lower lipase activity. For example, the lipase production of strains Pr-1 and Pr-5 exceeded the threshold for the probiotic strains and did not progress any further. Thus, by quantifying lipase production in sessile P. acnes cells, it was possible to screen among probiotic strains and select lead candidates with the most consistently low levels of lipase activity.

[0323] Thus, the pathogenic and probiotic strains secreted similar amounts of lipase in planktonic culture, whereas in attached culture the probiotic strains secreted much less lipase than the pathogenic strains. Figure 8 shows that the top probiotic candidates had lower lipase profiles compared to the pathogenic strains.

[0324] Testing the immunogenic potential of probiotic P. acnes strains: cell attachment The available pathogenic strains were identified to have the adhesion (tad) locus, which plays a role in the virulence of other mammalian pathogens [J Bacteriol (2000) 182:6169-6176; Nat Rev Microbiol (2007) 5:363-375; PNAS (2003) 100:7295-7300]. Greater adhesion to host cells may enhance virulence or induce an inflammatory host response. The probiotic strains were previously genotypically established not to contain the tad locus, and therefore were expected to adhere less tightly to epithelial cells. To assess whether there were any obvious differences in adhesion, the adhesion of the pathogenic and probiotic strains to A-431 skin epithelial cells was compared. Figure 9 shows that the top three probiotic candidates adhered less tightly to epithelial cells than the pathogenic strains. Interestingly, subtle but significant differences in cell adhesion were again found among different strain families of P. acnes, such that the P. acnes strain with the ProI ribosequence showed slightly higher cell adhesion (Pr-2 in Fig. 9), whereas the ProII strains adhered less tightly to cells (Pr-B, Pr-C in Fig. 9).

[0325] Comparison of virulence and probiotic P. acnes in a mouse ear inflammation model Having demonstrated the low immunogenic potential of the probiotic strains, showing lower production of lipase and less tight adhesion to epithelial cells, the inflammatory response of these strains was tested in the mouse ear inflammation model, an established model previously used to evaluate the proinflammatory potential of P. acnes in the context of acne. The proinflammatory potential of pathogenic and probiotic strains was compared in the following study.10 10 cfu strains were injected into the ears of CBA / J mice. Cohorts of 5 mice were assigned to each strain. Five days later, ears were excised and examined for inflammation. Levels of several inflammatory cytokines (IL-1β, IL-6, IL-17, TNFα) were measured and tissue sections were examined by histology. Figure 10 shows that the pathogenic strain had significantly higher levels of IL-1β, IL-6, IL-17 and TNFα compared to the probiotic strain.

[0326] Acute cutaneous safety and toxicity of probiotic strains in a minipig skin model The mini-pig model was used to test the probiotic strains for skin irritation. Pigs are one of the major animals used in translational research, and pig skin is physiologically, anatomically, biochemically, and immunologically similar to human skin. Mini-pigs are particularly commonly used to model human skin diseases and conditions, such as acne [Vet Pathol (2012) 49:344-356]. The probiotic strains were administered in two doses, 10 mL, to separate skin areas of the skin of three separate mini-pigs. 8 cfu and 10 9 cfu. Animals were observed daily for clinical signs and the skin at the administration site was scored using the Draize scoring system pre-dose, 0.5, 1, 4, 8 and 24 hours after dose administration. No erythema or edema was associated with the lead probiotic strain during the entire period (Table 1), and a Draize score of 0 was observed throughout. This demonstrates the safety of our probiotic strains upon acute exposure in the animal skin model.

[0327] Table 1: Acute skin safety / toxicity in the minipig skin model shows a good safety profile of the probiotic strains. The probiotic strains were administered to delimited skin areas in three male minipigs in normal (10 8 cfu) and acute (10 9 cfu) and were monitored for 24 hours after application. Erythema and edema were quantified using the Draize scoring system. The Draize score provides a relative severity of erythema and edema. A Draize score of 0 indicates the complete absence of erythema and edema, but was observed in all skin areas throughout the monitoring period.

[0328] [Table 1]

[0329] Example 3: Stability of bacteriophages in compositions with anti-acne compounds To determine whether the phage was stable in combination with salicylic acid or benzoyl peroxide (BPO), the phage was co-incubated with these drugs at low and high concentrations. The concentration range was determined by the permitted concentrations of these drugs as specified in the U.S. Food and Drug Administration (FDA) acne monographs for over-the-counter drugs. For salicylic acid, this is 0.5%-2% (w / v), and for BPO, the range is 2.5%-10% (w / v). Phage buffer was added to these drugs and their stability at 4°C was tested over a period of 60-90 days. Figure 15 shows that the phage was stable in the presence of both low and high doses of salicylic acid. In contrast, Figure 16 shows that benzoyl peroxide destabilized the phage and the observed rate of decrease in phage viability was steeper at higher concentrations of BPO.

[0330] Example 4 (hypothetical example) Treatment with a combination of bacteriophage and salicylic acid A double-blind, placebo-controlled study of a composition containing Propionibacterium acnes bacteriophage and salicylic acid is conducted to determine the comparative efficacy of this treatment with placebo, Propionibacterium acnes bacteriophage alone, and salicylic acid alone. Concentrations of 0.5% and 2% (w / v) salicylic acid are administered with and without Propionibacterium acnes bacteriophage. In all conditions containing Propionibacterium acnes bacteriophage, the phage is administered at 10 mg / dose. 9 The doses are in pfu (plaque forming units). Ten subjects with relatively severe acne will be treated in the following groups: (i) Placebo (no active drug) (ii) 0.5% salicylic acid as the only active agent (iii) 2% salicylic acid as the only active agent (iv) Propionibacterium acnes bacteriophage as the only active agent (v) 0.5% salicylic acid in combination with Propionibacterium acnes bacteriophage (in a single composition) (vi) 2% salicylic acid in combination with Propionibacterium acnes bacteriophage (in a single composition)

[0331] The combination of Propionibacterium acnes bacteriophage and salicylic acid achieves more than additive, i.e., synergistic, effects in treating acne (the combined effect of bacteriophage and salicylic acid is greater than the sum of the effects of the bacteriophage and salicylic acid when each agent is used separately). Efficacy of treatment is measured using lesion counts and Investigator Global Assessment (IGA) scores.

[0332] [Example 5] Hypothetical Example) Treatment using a combination of bacteriophage and sulfur A double-blind, placebo-controlled study of a composition containing Propionibacterium acnes bacteriophage and sulfur is conducted to determine the comparative efficacy of this treatment using placebo, Propionibacterium acnes bacteriophage alone, and sulfur alone. Sulfur concentrations of 3% and 10% (w / v) are administered with and without Propionibacterium acnes bacteriophage. In all conditions containing Propionibacterium acnes bacteriophage, the phage was administered at 10 mg / dose. 9 pfu. Ten subjects with relatively severe acne will be treated in the following groups: (i) Placebo (no active drug) (ii) 3% sulfur as the only active agent (iii) 10% sulfur as the only active agent (iv) Propionibacterium acnes bacteriophage as the only active agent (v) 3% sulfur and Propionibacterium acnes bacteriophage in a single composition. (vi) 10% sulfur in combination with Propionibacterium acnes bacteriophage (in a single composition)

[0333] The combination of Propionibacterium acnes bacteriophage and sulfur achieves more than additive, i.e., synergistic, effects in treating acne (the combined effect of bacteriophage and sulfur is greater than the sum of the effects of bacteriophage and sulfur when each agent is used separately). Efficacy of treatment is measured using lesion counts and Investigator Global Assessment (IGA) scores.

[0334] Example 6 (Prophetic Example) Treatment with a Combination of Bacteriophage and Benzoyl Peroxide A double-blind, placebo-controlled study of a composition containing Propionibacterium acnes bacteriophage and BPO is conducted to determine the comparative efficacy of this treatment with placebo, Propionibacterium acnes bacteriophage alone, and BPO alone. BPO at concentrations of 2.5% and 10% (w / v) is administered with and without Propionibacterium acnes bacteriophage. In all conditions containing Propionibacterium acnes bacteriophage, the phage was administered at a dose of 10 9 pfu. Ten subjects with relatively severe acne will be treated in the following groups: (i) Placebo (no active drug) (ii) 2.5% BPO as the only active agent (iii) 10% BPO as the only active agent (iv) Propionibacterium acnes bacteriophage as the only active agent (v) 2.5% BPO combined with Propionibacterium acnes bacteriophage (in a single composition) (vi) 10% BPO in combination with Propionibacterium acnes bacteriophage (in a single composition)

[0335] The combination of Propionibacterium acnes bacteriophage and BPO achieves more than additive, i.e. synergistic, effects in treating acne (the combined effect of bacteriophage and BPO is greater than the sum of the effects of bacteriophage and BPO when each agent is used separately). Efficacy of treatment is measured using lesion counts and Investigator Global Assessment (IGA) scores.

[0336] Example 7 (hypothetical example) Assay using a combination of bacteriophage and benzoyl peroxide In vitro studies are conducted to determine the effectiveness of (i) BPO; (ii) Propionibacterium acnes bacteriophage; or (iii) Propionibacterium acnes bacteriophage plus BPO in killing planktonic and sessile pathogenic P. acnes bacteria.

[0337] The combination of Propionibacterium acnes bacteriophage and BPO achieves a more than additive, i.e., synergistic, effect in killing sessile pathogenic P. acnes bacteria (the combined effect of bacteriophage and BPO is greater than the sum of the effects of bacteriophage and BPO when each agent is used separately). The keratolytic action of BPO (similar to salicylic acid and retinoids) helps the phage penetrate skin pores to access P. acnes deep within the pores.

Claims

1. A composition formulated for topical administration, comprising: The composition comprises: (a) PHIT-101 bacteriophage; (b) the enzyme dispersin, and (c) probiotic bacteria comprising a Propionibacterium acnes (P. acnes) strain; The P. acnes strain comprises a 16S rDNA sequence comprising SEQ ID NO:3 and SEQ ID NO:

4.

2. The composition described in claim 1, wherein the PHIT-101 bacteriophage comprises sequence number 1.

3. The composition described in claim 1, wherein the dispersin enzyme includes dispersin B.

4. The composition described in claim 3, wherein dispersin B includes sequence number 12.

5. The composition of claim 1, further comprising salicylic acid.

6. The composition described in claim 2, wherein the dispersin enzyme comprises sequence number 12 and the composition further comprises salicylic acid.

7. A method for treating acne in a subject in need of treatment, comprising topically administering the composition of claim 1 to an area of ​​the subject's skin containing acne.

8. The method of claim 7, comprising pretreating the area of ​​the subject's skin with a composition containing benzoyl peroxide prior to topical administration.

9. The method of claim 7, wherein the benzoyl peroxide composition is washed from the area of ​​the subject's skin prior to topical administration of the composition.

10. A method of treating acne in a subject in need thereof, comprising: The method comprises: (a) topically administering to an area of ​​the subject's skin containing acne a composition comprising a dispersin enzyme; (b) topically administering to an area of ​​the subject's skin that contains acne a composition comprising a PHIT-101 bacteriophage; and (c) topically administering to an area of ​​the subject's skin containing acne a composition comprising a Propionibacterium acnes (P. acnes) strain; The method, wherein the P. acnes strain comprises a 16S rDNA sequence comprising SEQ ID NOs: 3 and 4.

11. The method described in claim 10, wherein the dispersin enzyme includes dispersin B.

12. The method of claim 11, wherein the PHIT-101 bacteriophage comprises SEQ ID NO:

1.

13. The method described in claim 12, wherein the dispersin B enzyme comprises sequence number 12.

14. The method of claim 10, wherein prior to step (a), the area of ​​the subject's skin containing acne is pre-treated with benzoyl peroxide.

15. The method of claim 14, wherein prior to step (a), benzoyl peroxide is removed.

16. The method of claim 10, further comprising: (d) topically administering to the area of ​​the subject's skin containing acne a composition comprising one or more of salicylic acid, resorcinol, and sulfur.

17. The method of claim 10, wherein the composition comprising the bacteriophage further comprises salicylic acid.

18. The method described in claim 10, wherein the composition containing the dispersin enzyme and the composition containing the bacteriophage are the same.

19. The method described in claim 10, wherein the composition containing the bacteriophage and the composition containing the probiotic bacteria are the same.

20. The method of claim 19, wherein the composition comprising the bacteriophage and the probiotic bacteria further comprises salicylic acid.

Images