Bismuth-thiol compositions and methods for treating wounds

Abstract

To provide a composition useful for the treatment of diabetic foot infections (DFI) and topical wounds resulting therefrom.SOLUTION: There is provided a topical wound treatment composition, comprising a plurality of microparticles of a therapeutically effective amount of a bismuthiol (BT) compound suspended therein, wherein the D90 of the microparticles is less than about 2 μm, and wherein the BT compound is selected from the group consisting of BisBAL, BisEDT, Bis- dimercaprol, BisDTT, Bis-2- mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, BisPyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, bismuth-1-mercapto-2-propanol, and Bis-EDT / 2-hydroxy-1-propanethiol, and wherein the composition is applied to the wound.SELECTED DRAWING: Figure 5A

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 62 / 712,555, filed July 31, 2018, and U.S. Provisional Patent Application No. 62 / 820,006, filed March 18, 2019, the contents of which are incorporated by reference herein in their entireties. [Background technology]

[0002] Diabetic foot infections (DFIs) are a frequent and serious complication of diabetes mellitus (DM) and a worldwide cause of non-traumatic lower limb amputations (Jeffcoate WJ, et al. 2003, Lancet, 361:1545-1551). In current clinical practice, treatment of DFIs includes debridement and systemic antibiotics (see, e.g., Lipsky BA, et al. 2004, Clin Infect Dis. 39:885-910). Nevertheless, due to the lack of vascularization and the local microenvironment, antibiotic concentrations are many times subtherapeutic (Lipsky BA, et al. 2009, Clin Infect Dis. 49:1541-1549). Furthermore, the increasing occurrence of multidrug-resistant bacteria such as methicillin-resistant Staphylococcus aureus, as well as pan-drug-resistant non-fermenting negative bacilli, threatens the outcomes of an increasing number of community-based and hospitalized patients (Mendes JJ, et al. 2012, Diabetes Res Clin Pract. 95(1):153-161; Tascini C, et al. 2011, Diabetes Res Clin Pract. 94(1):133-139). Thus, there remains a need to identify new strategies for the treatment, control, and management of DFIs. Summary of the Invention

[0003] In some embodiments, the present disclosure provides a method of treating a local wound comprising administering to a subject in need thereof a therapeutically effective amount of a composition comprising a bismuth thiol compound, wherein the composition is applied to the infection (e.g., applied to the skin and tissues on and / or surrounding the infection and / or to the wound itself). The local wound may be a skin ulcer (e.g., a skin ulcer of the lower leg). In some embodiments, the skin ulcer is one or more of a foot ulcer, a diabetic foot ulcer, an ischemic ulcer, a gangrenous ulcer, a venous stasis ulcer, a decubitus ulcer, a Buruli ulcer, or a traumatic ulcer. In some embodiments, the local wound is infected with one or more bacterial and / or fungal pathogens. In some embodiments, the local wound is a diabetic foot ulcer. In some embodiments, the diabetic foot ulcer is a diabetic foot ulcer infection.In some embodiments, the local wound is free of the following bacterial pathogens: Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp., Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, Coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium acnes, S.dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas Sp.

[0004] In some embodiments of the methods of treating a topical wound, the subject experiences one or more of the following results after completion of dosing: a) the wound is healed or substantially healed within 12 weeks of the initial administration of the composition; and / or b) preventing amputations and / or infection-related surgery; and / or c) the wound is partially or completely closed; and / or d) the wound is reduced in size from approximately 1% reduction in size compared to the original wound size to complete disappearance of the wound; and / or e) The wound is 30 days or older (i.e., at the start of treatment) and is healed or substantially healed. In some embodiments, the subject experiences two or more of the recited results. In some embodiments, the subject experiences three or more of the recited results. In some embodiments, the subject experiences four or more of the recited results. In some embodiments, the subject experiences all of the recited results.

[0005] In some embodiments of the method for treating a topical wound, the BT composition comprises one or more BT compounds selected from BisBAL, BisEDT, Bis-dimercaprol, BisDTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, BisPyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, bismuth-1-mercapto-2-propanol, and bis-EDT / 2-hydroxy-1-propanethiol. In some embodiments, the BT compound is BisEDT. In some embodiments, the composition is a suspension of microparticles comprising the BT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

[0006] In some embodiments of the method of treating a topical wound, the BT composition comprises BisEDT, and the BisEDT applied is about 20 μg / cm 2In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0007] In some embodiments of the method of treating a topical wound, following administration of the BT composition, one or more of the following occurs: (i) reducing and / or dispersing microbial (e.g., bacterial and / or fungal) biofilms, (ii) impairing the growth or formation of microbial (e.g., bacterial and / or fungal) biofilms, and (iii) preventing the reformation or spread of microbial (e.g., bacterial and / or fungal) biofilms. In some embodiments, the BT composition treats, manages, and / or reduces the severity of a diabetic foot infection by one or both of (i) preventing infection by bacterial or fungal pathogens and / or (ii) reducing bacterial or fungal pathogens. In some embodiments, the BT composition treats, manages, or reduces the severity of an infection by one or more of: (i) preventing the production or secretion of exotoxins from bacterial or fungal pathogens; (ii) inhibiting cell viability or cell proliferation of planktonic cells of bacterial or fungal pathogens; (iii) inhibiting biofilm formation by bacterial or fungal pathogens; (iv) inhibiting the invasiveness of biofilms or microbial pathogens into underlying tissues (e.g., subcutaneous tissue); (v) inhibiting the virulence of biofilms or microbial pathogens into underlying tissues (e.g., subcutaneous tissue); (vi) inhibiting biofilm viability or biofilm growth of biofilm-forming cells of bacterial or fungal pathogens; and / or (vii) preventing biofilm reformation after debridement.

[0008] In some embodiments of the method of treating a topical wound, the BT composition administered is 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~Approx. 10,000μg / cm 2In some embodiments, the administered BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~about 200μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 250 μg / cm 2 ~Approx. 5,000μg / cm 2 is present on the surface at a concentration of

[0009] In some embodiments of the method for treating a local wound, the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months. In some embodiments, the BT composition is administered once a day or three times a week. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of time ranging from about 1 week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of time ranging from about 4 weeks to about 10 weeks.

[0010] In some embodiments of the method of treating a topical wound, the wound area is about 0.1 cm 2 ~about 250cm 2 In some embodiments of the method of treating a localized wound, the wound area is about 250 cm 2 ~about 500cm 2 For example, the wound area may be the wound area of ​​one wound or more than one wound. In such an embodiment, the total wound area is 500 cm 2 It may be much larger.

[0011] In some embodiments, the present disclosure provides a method of treating a microbial (e.g., bacterial and / or fungal) infection comprising administering to a subject in need thereof a therapeutically effective amount of a composition comprising a bismuth thiol compound, wherein the composition is applied to the infection (e.g., applied to the surface of the infection). In some embodiments, the microbial (e.g., bacterial and / or fungal) infection is a diabetic foot infection.In some embodiments, the microbial (e.g., bacterial and / or fungal) infection is caused by the following bacterial pathogens: Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp., Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium acnes, S.dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis. In some embodiments, the infection is associated with a wound (eg, an ulcer) and the subject experiences one or more of the following outcomes after completion of the medication: a) the wound is healed or substantially healed within 12 weeks (e.g., within 4 weeks) of the initial administration of the composition; and / or b) preventing amputations and / or infection-related surgery; and / or c) the wound is partially or completely closed; and / or d) the wound is reduced in size from approximately 1% reduction in size compared to the original wound size to complete disappearance of the wound; and / or e) The wound is at least 30 days old and is healed or substantially healed. In some embodiments, the subject experiences two or more of the recited results. In some embodiments, the subject experiences three or more of the recited results. In some embodiments, the subject experiences four or more of the recited results. In some embodiments, the subject experiences all of the recited results.

[0012] In some embodiments of the method of treating a microbial (e.g., bacterial and / or fungal) infection, the BT composition comprises one or more BT compounds selected from BisBAL, BisEDT, bis-dimercaprol, BisDTT, bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, BisPyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, bismuth-1-mercapto-2-propanol, and bis-EDT / 2-hydroxy-1-propanethiol. In some embodiments, the BT compound is BisEDT. In some embodiments, the composition is a suspension of microparticles comprising the BT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

[0013] In some embodiments of the method of treating a microbial (e.g., bacterial and / or fungal) infection, the BT composition comprises BisEDT, and the BisEDT applied is about 20 μg / cm 2 present on surfaces in concentrations exceeding

[0014] In some embodiments of the method of treating a microbial (e.g., bacterial and / or fungal) infection, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4.

[0015] In some embodiments of the method of treating a microbial (e.g., bacterial and / or fungal) infection, the method comprises at least one of (i) reducing and / or dispersing a microbial (e.g., bacterial and / or fungal) biofilm, (ii) impairing the growth or formation of a microbial (e.g., bacterial and / or fungal) biofilm, and (iii) preventing the reformation or spread of a microbial (e.g., bacterial and / or fungal) biofilm. In some embodiments, the BT composition treats, manages, or reduces the severity of a diabetic foot infection by one or both of (i) preventing infection by bacterial or fungal pathogens and (ii) reducing bacterial or fungal pathogens. In some embodiments, the BT composition treats, manages, or reduces the severity of an infection by one or more of: (i) preventing the production or secretion of exotoxins from bacterial or fungal pathogens; (ii) inhibiting cell viability or cell proliferation of planktonic cells of bacterial or fungal pathogens; (iii) inhibiting the formation of biofilms or microbial pathogens by bacterial or fungal pathogens; (iv) inhibiting the invasiveness of biofilms into underlying tissues (e.g., subcutaneous tissue); (v) inhibiting the virulence of biofilms or microbial pathogens on underlying tissues (e.g., subcutaneous tissue); (vi) inhibiting biofilm viability or biofilm growth of biofilm-forming cells of bacterial or fungal pathogens; and / or (vii) preventing the reformation of biofilms after debridement.

[0016] In some embodiments of the methods of treating a microbial (e.g., bacterial and / or fungal) infection, the applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm2 ~Approx. 10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 250 μg / cm 2 ~Approx. 5,000μg / cm 2 is present on the surface at a concentration of

[0017] In some embodiments of the method of treating a microbial (e.g., bacterial and / or fungal) infection, the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every other month. In some embodiments, the BT composition is administered once a day or three times a week. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 10 weeks.

[0018] In some embodiments of the methods of treating a microbial (e.g., bacterial and / or fungal) infection, the wound area is about 0.1 cm 2 ~about 250cm 2 In some embodiments of the methods of treating a microbial (e.g., bacterial and / or fungal) infection, the wound area is about 250 cm 2 ~about 500cm 2 The wound area may be the wound area of ​​one wound or more than one wound.

[0019] In some embodiments, the disclosure provides a method of healing a wound in a subject having a diabetic foot infection comprising administering to the subject a therapeutically effective amount of a composition comprising BisEDT, wherein the composition is a suspension of microparticles comprising said BisEDT, at least 70% of the microparticles having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm, the composition is applied to the infection (e.g., applied to the surface of the infection), and the wound is healed or substantially healed within 12 weeks of initial administration of the composition. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0020] In some embodiments of the method of healing a wound in a subject with a diabetic foot infection, the BT composition applied is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~Approx. 10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 is present on the surface at a dose of

[0021] In some embodiments of the method for healing a wound of a subject with diabetic foot infection, the BT composition is administered 3 times a day, 2 times a day, once a day, every other day, once every 3 days, 3 times a week, once a week, once every other week, once a month, or once every other month. In some embodiments, the wound is healed 4 weeks, 8 weeks, or 12 weeks after the first administration of the BT composition. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of time ranging from about 1 week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of time ranging from about 4 weeks.

[0022] In some embodiments of the method of healing a wound in a subject with a diabetic foot infection, the wound area is about 0.1 cm 2 ~about 250cm 2 In some embodiments, the wound area is about 250 cm 2 ~about 500cm 2 The wound area may be the wound area of ​​one wound or more than one wound.

[0023] In some embodiments, the disclosure provides a method of reducing the risk of amputation and / or infection-related surgery in a subject having a diabetic foot infection comprising administering to the subject a therapeutically effective amount of a composition comprising BisEDT, wherein the composition is applied to the infection (e.g., applied to the surface of the infection), and the risk of amputation and / or infection-related surgery is reduced by about 1% to about 100% compared to a similarly situated subject not treated with a therapeutically effective amount of a composition comprising a bismuth thiol compound. In some embodiments, the composition is a suspension of microparticles comprising said BisEDT having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4.

[0024] In some embodiments, the method of reducing the risk of amputation and / or infection-related surgery in a subject with a diabetic foot infection comprises administering about 1 μg / cm of the BT composition. 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 is present on the surface at a dose of

[0025] In some embodiments, the method of reducing the risk of amputation and / or infection-related surgery in a subject with a diabetic foot infection comprises administering the BT composition three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every other month. In some embodiments, the BT composition is administered once a day or three times a week. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of time ranging from about one week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of time ranging from about one week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of time ranging from about four weeks.

[0026] In some embodiments, the method of reducing the risk of amputation and / or infection-related surgery in a subject with a diabetic foot infection comprises administering to a subject a wound having an area of ​​about 0.1 cm 2 ~about 250cm 2 In some embodiments, the wound area is about 250 cm 2 ~about 500cm 2 The wound area may be the wound area of ​​one wound or more than one wound.

[0027] In some embodiments, the disclosure provides a method of closing a wound in a subject having a diabetic foot infection, comprising administering to the subject a therapeutically effective amount of a composition comprising BisEDT. In some embodiments, the composition is a suspension of microparticles comprising said BisEDT, wherein at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4.

[0028] In some embodiments, in the method of closing a wound in a subject with a diabetic foot infection, the applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~Approx. 10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 is present on the surface at a dose of

[0029] In some embodiments, the method of closing a wound in a subject with a diabetic foot infection comprises applying the composition to the infection (e.g., applying to the surface of the infection) and closing the wound within 12 weeks of the first administration of the composition. In some embodiments, the BT composition is administered once a day or three times a week. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of time ranging from about 1 week to about 12 weeks ...4 weeks.

[0030] In some embodiments, the method of closing a wound in a subject with a diabetic foot infection comprises: 2 ~about 250cm 2 In some embodiments, the wound area is about 250 cm 2 ~about 500cm 2 The wound area may be the wound area of ​​one wound or more than one wound.

[0031] In some embodiments, the disclosure provides a method for wound size reduction in a subject with a diabetic foot infection, comprising administering to the subject a therapeutically effective amount of a composition comprising BisEDT, wherein the composition is applied to the infection (e.g., applied to the surface of the infection), and within 12 weeks of the initial administration of the composition, the wound is reduced in size from about 1% reduction compared to the original wound size to complete clearance of the wound. In some embodiments, the wound is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%. In some embodiments, the wound is reduced by about 50%.

[0032] In some embodiments, the method for wound size reduction in a subject with a diabetic foot infection comprises a composition comprising a suspension of microparticles comprising BisEDT, wherein at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0033] In some embodiments, in the method for reducing wound size in a subject with a diabetic foot infection, the applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~Approx. 10,000μg / cm 2In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 is present on the surface at a dose of

[0034] In some embodiments, the method for wound size reduction in a subject with diabetic foot infection is to administer the BT composition three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months. In some embodiments, the BT composition is administered once a day or three times a week. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of time ranging from about 1 week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of time ranging from about 4 weeks.

[0035] In some embodiments, the method for reducing wound size in a subject with a diabetic foot infection comprises administering to a subject a wound having an area of ​​about 0.1 cm 2 ~about 250cm 2 In some embodiments, the wound area is about 250 cm 2 ~about 500cm 2 The wound area may be the wound area of ​​one wound or more than one wound. In some embodiments, the wound surface area of ​​the wound is reduced by 50% 12 weeks after the first administration of the BT composition, the BT composition being BisEDT. In some embodiments, the wound surface area of ​​the wound is reduced by 50% 4 weeks after the first administration of the BisEDT composition. In some embodiments, the wound surface area is measured using digital photography or manual measurement.

[0036] In some embodiments, the present disclosure provides a method for preventing amputation and / or infection-related surgery in a subject with a diabetic foot infection, comprising administering to the subject a therapeutically effective amount of a BT composition. In some embodiments, the BT composition is a suspension of microparticles comprising the BisEDT, wherein at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4.

[0037] In some embodiments, in the method of preventing amputation and / or infection-related surgery in a subject with a diabetic foot infection, the applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 is present on the surface at a dose of

[0038] In some embodiments, the disclosure provides a method of treating a wound in a subject, wherein the wound is 30 days or older, comprising administering a therapeutically effective amount of a BT composition to the subject. In some embodiments, the subject has a diabetic foot infection. In some embodiments, the BT composition is a suspension of microparticles comprising BisEDT, wherein at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4.

[0039] In some embodiments, the disclosure provides a method of treating a wound in a subject, wherein the wound is within 30 days, comprising administering a therapeutically effective amount of a BT composition to the subject. In some embodiments, the wound is a diabetic foot infection. In some embodiments, the BT composition is a suspension of microparticles comprising BisEDT, wherein at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate. For example, the wound may be about the 1st day, about the 2nd day, about the 3rd day, about the 4th day, about the 5th day, about the 6th day, about the 7th day, about the 8th day, about the 9th day, about the 10th day, about the 11th day, about the 12th day, about the 13th day, about the 14th day, about the 15th day, about the 16th day, about the 17th day, about the 18th day, about the 19th day, about the 20th day, about the 21st day, about the 22nd day, about the 23rd day, about the 24th day, about the 25th day, about the 26th day, about the 27th day, about the 28th day, about the 29th day, or about the 30th day.

[0040] In some embodiments, in the method of treating a wound in a subject, the wound is 30 days or older, the BT composition applied is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm2 ~Approx. 10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 is present on the surface at a dose of

[0041] In some embodiments of the method of treating a wound in a subject, the wound is more than 30 days old, the wound is more than 2 months old, more than 3 months old, more than 4 months old, more than 5 months old, more than 6 months old, more than 7 months old, more than 8 months old, more than 9 months old, more than 10 months old, more than 11 months old, or more than 1 year old. In some embodiments, the wound is more than 2 months old. In some embodiments, the wound is more than 3 months old.

[0042] In some embodiments of the present disclosure, the BT composition described herein comprises BisEDT, and the composition is a suspension of microparticles comprising said BisEDT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, at least 70% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm. In some embodiments, the VMD is about 0.4 μm to about 3 μm. In some embodiments, the VMD is about 1.0 μm to about 2.0 μm. In some embodiments, at least 70%, 75%, 80%, 90%, or 95% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm. In some embodiments, at least 80% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm. In some embodiments, at least 90% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm. In some embodiments, substantially all of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm.In some embodiments, the VMD is about 0.4 μm to about 3 μm. In some embodiments, the VMD is about 1.0 μm to about 2.0 μm. In some embodiments, 90% of the particles are smaller than about 10 μm. For example, in some embodiments, 90% of the particles are smaller than about 10 μm, 9 μm, 8 μm, 7 μm, 6 μm, 5 μm, 4 μm, 3 μm, 2 μm, or about 1 μm. [Brief description of the drawings]

[0043] [Figure 1] FIG. 1 shows that BisEDT at 2500 μg / mL reduces the Wound Actual Index by approximately 5 logs compared to diluent. [Diagram 2] FIG. 1 shows that 2.5 mg / mL BisEDT completely inhibits splenic infection (greater than 4-fold log reduction) compared to diluent. [Diagram 3] FIG. 10 mg / mL BisEDT reduced CFU by over 8 logs (complete inhibition compared to untreated control) and 5 mg / mL BisEDT reduced CFU by 1.5-2 logs. [Figure 4] FIG. 1 shows that BisEDT and Prontosan at 5 mg / mL and 10 mg / mL completely inhibit splenic CFU compared to untreated controls (greater than 3.5 log reduction). [Figure 5A] Bioluminescence images of mice treated with 10 mg / mL BisEDT are shown. [Figure 5B] Bioluminescence images of mice treated with 5 mg / mL BisEDT are shown. [Figure 5C] Bioluminescence images of mice treated with 2.5 mg / mL BisEDT are shown. [Figure 5D] Bioluminescence images of mice treated with 0.75 mg / mL BisEDT are shown. [Figure 5E] Bioluminescence images of mice treated with Prontosan are shown. [Figure 5F] Bioluminescence images of untreated mice are shown in Figure 5F. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0044] definition Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art of this disclosure. The following references: Singleton et al., Dictionary of Microbiology and Molecular Biology, (2nd ed. 1994); The Cambridge Dictionary of Science and Technology, (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology, (1991) provide those of ordinary skill in the art with general definitions of many of the terms used in this disclosure. As used herein, the following terms have the meanings ascribed to them below, unless otherwise specified.

[0045] As used herein, the verb "comprise" and its conjugations as used in the description and claims are used in an open-ended sense meaning that items following the word are included, but items not specifically mentioned are not excluded. The disclosure preferably "comprises," "consists of," or "consists essentially of" the steps, elements, and / or reagents recited in the claims.

[0046] Unless specifically stated or clear from context, as used herein, the term "or" is understood to be inclusive. Unless specifically stated or clear from context, as used herein, the terms "a," "an," and "the" are understood to be singular or plural.

[0047] Throughout this specification, the terms "about" and / or "approximately" may be used in conjunction with numerical values ​​and / or ranges. The term "about" is understood to mean a value close to the recited value. Additionally, the phrases "less than about [value]" or "greater than about [value]" should be understood in light of the definition of the term "about" provided herein. The terms "about" and "approximately" may be used interchangeably.

[0048] An "alkyl" group or "alkane" is a straight or branched chain non-aromatic hydrocarbon that is fully saturated. Typically, a straight or branched chain alkyl group has from 1 to about 20 carbon atoms, e.g., from 1 to about 10, unless otherwise defined. Examples of straight and branched chain alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl, and octyl. C1-C6 straight or branched chain alkyl groups are also referred to as "lower alkyl" groups.

[0049] Furthermore, the term "alkyl" (or "lower alkyl"), as used throughout the specification, examples, and claims, is intended to include both "unsubstituted alkyl" and "substituted alkyl," the latter of which refers to an alkyl moiety having a substituent in place of a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents can include, for example, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties, unless otherwise specified. It will be understood by those skilled in the art that moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate. For example, substituents of substituted alkyl can include amino, azido, imino, amido, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl, and sulfonates), and silyl groups, as well as substituted and unsubstituted forms of ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, and the like. Exemplary substituted alkyls are described below. Cycloalkyls can be further substituted with alkyls, alkenyls, alkoxys, alkylthios, aminoalkyls, carbonyl-substituted alkyls, -CF3, -CN, and the like.

[0050] "C x-y The term "alkyl" when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups containing x to y carbons in the chain. For example, "C x-yThe term "alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups including straight chain alkyl and branched chain alkyl groups containing from x to y carbons in the chain, including haloalkyl groups such as, for example, trifluoromethyl and 2,2,2-trifluoroethyl. C0 alkyl represents a hydrogen if the group is in a terminal position or a bond if it is internal. 2-y alkenyl" and "C 2-y "Alkenyl" refers to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one double or triple bond respectively.

[0051] The term "alkylamino," as used herein, refers to an amino group substituted with at least one alkyl group.

[0052] The term "alkylthio," as used herein, refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

[0053] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines, and salts thereof, e.g.,

[0054] [ka] wherein each R 31 each independently represents a hydrogen or a hydrocarbyl group, or two R 31 together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure. The term "aminoalkyl" as used herein refers to an alkyl group substituted with an amino group.

[0055] The term "aryl" as used herein includes substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. In some embodiments, the ring is 5- to 7-membered, e.g., 6-membered. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, at least one of which is aromatic, e.g., the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0056] The term "bismuth" refers to the 83rd element of the periodic table, or an atom or ion thereof. Bismuth can exist in a metallic state or an ionized state, e.g., oxidation states III or V. Bismuth ions can be complexed with anions to make bismuth salts, or to form complex anions that are then further complexed with one or more additional cation(s). Bismuth can also form covalent bonds with other atoms, such as sulfur.

[0057] As disclosed herein, a "bismuth thiol compound" or "BT compound" is a compound having a bismuth atom covalently bonded to one, two, or three other sulfur atoms present in one or more thiol compounds. The term "thiol" refers to a carbon-containing compound or fragment thereof containing an -SH group and can be represented by the general formula R-SH. These thiol compounds include compounds with one, two, three, or more S atoms. The thiol compounds can have other functionalities such as, for example, alkyl, hydroxyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, amino, and other substituents. Thiol compounds with two or more S atoms can chelate a bismuth atom, so that two S atoms from the same molecule are covalently bonded to a bismuth atom. Exemplary bismuth thiol compounds are shown below:

[0058] [ka] The terms "carbocycle" and "carbocyclic" as used herein refer to a saturated or unsaturated ring in which each atom of the ring is carbon. The term carbocycle includes both aromatic and non-aromatic carbocycles. Non-aromatic carbocycles include both cycloalkane rings in which all carbon atoms are saturated, and cycloalkene rings which contain at least one double bond.

[0059] The term "carbocycle" includes 5- to 7-membered monocyclic rings and 8- to 12-membered bicyclic rings. Each ring of a bicyclic carbocycle can be selected from saturated, unsaturated, and aromatic rings. Carbocycles include bicyclic molecules in which one, two, or more atoms are shared between the two rings. The term "fused carbocycle" refers to a bicyclic carbocycle in which each of the rings shares two adjacent atoms with the other ring. Each ring of a fused carbocycle can be selected from saturated, unsaturated, and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, can be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated, and aromatic bicyclic rings is included in the definition of carbocycle, if valence permits. Exemplary "carbocycles" include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene, and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. A "carbocycle" can be substituted at any one or more positions capable of carrying a hydrogen atom.

[0060] A "cycloalkyl" group is a cyclic hydrocarbon that is fully saturated. "Cycloalkyl" includes monocyclic and bicyclic rings. Typically, a monocyclic cycloalkyl group has from 3 to about 10 carbon atoms, more typically from 3 to 8 carbon atoms, unless otherwise defined. The second ring of a bicyclic cycloalkyl can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. Cycloalkyl includes bicyclic molecules in which one, two, or three or more atoms are shared between the two rings. The term "fused cycloalkyl" refers to a bicyclic cycloalkyl in which each of the rings shares two adjacent atoms with the other ring. The second ring of a fused bicyclic cycloalkyl can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. A "cycloalkenyl" group is a cyclic hydrocarbon that contains one or more double bonds.

[0061] The terms "halo" and "halogen" as used herein mean halogen and include chloro, fluoro, bromo, and iodo.

[0062] The terms "hetaralkyl" and "heteroaralkyl," as used herein, refer to an alkyl group substituted with a hetaryl group.

[0063] The term "heteroalkyl," as used herein, refers to a saturated or unsaturated chain of carbon atoms and at least one heteroatom, where no two heteroatoms are adjacent.

[0064] The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic ring structures, e.g., 5- to 7-membered rings, e.g., 5- to 6-membered rings, which ring structures include at least one heteroatom, e.g., 1 to 4 heteroatoms, e.g., 1 or 2 heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, at least one of which is heteroaromatic, e.g., the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, and the like.

[0065] The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0066] The terms "heterocyclyl", "heterocycle", and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures, e.g., 3- to 10-membered, e.g., 3- to 7-membered, rings, which contain at least one heteroatom, e.g., 1 to 4 heteroatoms, e.g., 1 or 2 heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, at least one of which is heterocyclic, e.g., the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

[0067] The term "heterocyclylalkyl," as used herein, refers to an alkyl group substituted with a heterocycle group.

[0068] The term "hydrocarbyl" as used herein refers to a group that does not have =O or =S substituents and that usually has at least one carbon-hydrogen bond and a primarily carbon backbone, but which may optionally contain heteroatoms, bonded through carbon atoms. Thus, groups such as methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered hydrocarbyl for the purposes of this application, while substituents such as acetyl (which has =O substituent on the bonded carbon) and ethoxy (which is bonded through an oxygen rather than a carbon) are not hydrocarbyl. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocycle, heterocyclyl, alkyl, alkenyl, alkynyl, and combinations thereof.

[0069] The term "lower" when used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups in which there are 10 or fewer, e.g., 6 or fewer, non-hydrogen atoms in the substituent. "Lower alkyl" refers, for example, to alkyl groups containing 10 or fewer, e.g., 6 or fewer, carbon atoms. In certain embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein are lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, respectively, whether they appear alone or in combination with other substituents, e.g., hydroxyalkyl and aralkyl in the enumeration (where, for example, atoms in an aryl group are not counted when counting carbon atoms in an alkyl substituent).

[0070] The terms "polycyclyl," "polycycle," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl) in which two or more atoms are common to two adjacent rings, e.g., the rings are "fused rings." Each of the rings of a polycycle can be substituted or unsubstituted. In certain embodiments, each ring of a polycycle contains 3 to 10 atoms in the ring, e.g., 5 to 7 atoms.

[0071] The term "N-oxide" refers to a zwitterionic group containing a nitrogen atom in the +1 oxidation state bonded to an oxygen atom in the -1 oxidation state. A non-limiting example of an N-oxide is the pyridinium N-oxide shown below. As used herein, the term "N-oxide" includes substituents of other groups with this functionality.

[0072] [ka] The term "substituted" refers to a moiety having a substituent replacing a hydrogen on one or more carbons of the backbone. It will be understood that "substituted" or "substituted with" includes the implicit proviso that such substitution results in a stable compound, e.g., a stable compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, and the like, subject to the permissible valences of the substituted atom and substituent. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, heteroatoms such as nitrogen can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of the heteroatom. The substituents may include any of the substituents described herein, such as halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. It will be understood by those skilled in the art that the substituents themselves may be substituted, where appropriate. Unless specifically stated as "unsubstituted," references to chemical moieties herein are understood to include substituted variants. For example, references to an "aryl" group or moiety implicitly include both substituted and unsubstituted variants.

[0073] The term "thioalkyl" as used herein refers to an alkyl group substituted with a thiol group. A "thiol compound" as discussed above can include a thioalkyl as a substituent on the compound structure. A thiol compound can have, for example, one, two, three, or more thioalkyl groups.

[0074] The term "thioether" as used herein is equivalent to ether, where the oxygen is replaced by sulfur.

[0075] The term "subject" to which administration is contemplated includes, but is not limited to, humans (i.e., male or female of any age, e.g., a pediatric subject (e.g., infant, child, adolescent) or an adult subject (e.g., young adult, middle-aged adult, or elderly adult)); and / or other primates (e.g., cynomolgus monkeys, rhesus monkeys); mammals, including, but not limited to, commercially relevant mammals such as cows, pigs, horses, sheep, goats, cats, and / or dogs; and / or birds, including commercially relevant birds such as chickens, ducks, geese, quail, and / or turkeys. A preferred subject is a human.

[0076] As used herein, the term "co-administration" refers to any form of administration of two or more different therapeutic compounds, such that a second compound is administered while a previously administered therapeutic compound is still effective in the body (e.g., two compounds are effective in a patient at the same time, which may include a synergistic effect of the two compounds). For example, different therapeutic compounds can be administered simultaneously or sequentially, either in the same formulation or in separate formulations. In certain embodiments, different therapeutic compounds can be administered within 1 hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, or within 1 week.

[0077] "Concurrent administration" refers to the administration of two drugs in any manner that allows the pharmacological effects of both drugs to occur simultaneously in a patient.Concurrent administration does not therefore require that a single pharmaceutical composition, the same dosage form, or even the same route of administration be used for both drugs, or that the two drugs be administered at exactly the same time.However, in some situations, concurrent administration will be most conveniently achieved by substantially the same time, the same dosage form, and the same route of administration.

[0078] As used herein, a therapeutic agent that "prevents" a disorder or condition refers to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in treated samples compared to untreated control samples, or delays the onset of or reduces the severity of one or more signs and symptoms of the disorder or condition compared to untreated control samples.

[0079] The term "treat" refers to one or more of alleviating, ameliorating, delaying, reducing, ameliorating, or managing at least one symptom of a condition in a subject. The term "treat" may also refer to one or more of preventing, delaying the onset (i.e., the period prior to clinical symptoms of a condition), or reducing the risk of developing or worsening a condition.

[0080] The term "managing" includes therapeutic treatment as defined above. Managing includes achieving a steady state level of infection as determined by methods known in the art. Steady state includes one or more assessments of the severity of the infection(s), the size and location of the infection(s), the number of different microbial pathogens present in the infection(s), the level of antibiotic-resistant or resistant microbial pathogens, the extent of response to treatment such as with the BT compositions disclosed herein, the extent of biofilm formation and reduction, and side effects experienced by the subject. During management of an infection, the infection may fluctuate in severity from increasing to decreasing, the amount or extent of infection, the amount of side effects experienced by the subject, or other indicia of subject outcome. Over a period of time such as days, months, or years, the extent of management of an infection can be determined by evaluation of the above factors to assess whether the clinical course of the infection has improved, is bacteriostatic, or has worsened. In some embodiments, managing an infection includes successful treatment of microbial pathogens that are otherwise drug-resistant or drug-resistant.

[0081] The term "reducing the severity" of an infection(s) refers to an improvement in the clinical course of the infection on a measurable basis. Such criteria can include measurable indicators such as a reduction in the extent of the infection(s), whether the infection(s) are considered acute, the number and endemicness of the microbial pathogen(s) causing the infection(s), the extent / spread / amount of microbial (e.g., bacterial and / or fungal) biofilm, and side effects experienced by the subject. In some embodiments, reducing the severity of an infection is determined by measuring an improvement in clinical signs and symptoms of the infection. In some embodiments, reducing the severity includes stopping a steady decline in outcomes to achieve stabilized infection(s), such that the subject enters into successful management of the infection(s). In other embodiments, reducing the severity can result in substantial to complete treatment of the infection(s).

[0082] In some embodiments, reducing the severity of the infection and / or symptoms can be related to patient reported outcomes ("PROs"). PRO instruments are defined as subjective health status measures elicited from the patient to determine "how the patient feels and functions in relation to their health status." PROs are particularly useful for reporting DFI outcomes and whether the severity of symptoms has been reduced or alleviated. Such symptoms can be observable events, behaviors, or emotions (e.g., ability to walk fast, lack of appetite, expression of anger), or unobservable outcomes known only to the patient (e.g., perception of pain, feelings of depression). In some embodiments, reducing the severity of the infection and / or symptoms can be determined by physician assessment as commonly known in the art, for example, by an 8-item wound score.

[0083] As used herein, "effective amount" refers to an amount that is sufficient to achieve a desired biological effect.As used herein, "therapeutically effective amount" refers to an amount that is sufficient to achieve a desired therapeutic effect.For example, a therapeutically effective amount can refer to an amount that is sufficient to improve at least one sign or symptom of an infectious disease.

[0084] As used herein, the phrase "pharmacologically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are within the scope of sound medical judgment and suitable for use in contact with the tissues of a subject without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0085] A "response" to a method of treatment can include, among other things, a reduction or amelioration of negative signs and symptoms, a decrease in the progression of the infection or its symptoms, an increase in beneficial symptoms or clinical outcomes, a reduction in side effects, stabilization of the infection, and, among other things, partial or complete cure of the infection, partial or complete wound closure, reduction in wound size, or complete or substantially complete re-epithelialization.

[0086] "Antibiotic susceptibility or sensitivity" refers to whether a bacterium is successfully treated by a given antibiotic. Similarly, "antifungal susceptibility or sensitivity" refers to whether a fungus is successfully treated by a given antibiotic. Susceptibility testing can be performed by methods known in the art, such as the Kirby-Bauer method, the Stokes method, and the agar broth dilution method. The effectiveness of an antibiotic in killing bacteria or preventing bacteria from growing can be observed as areas of reduced or stable amounts of bacterial growth, respectively, in media such as wafer cultures, agar cultures, or broth cultures.

[0087] "Antibacterial resistance" refers to the ability of microorganisms such as bacteria and fungi to naturally resist killing by antibiotics. It is caused not by mutant microorganisms, but rather by microbial cells that exist in a transient, dormant, non-dividing state. Antibiotic resistance or drug resistance is caused by a small subpopulation of microbial cells called persisters. Persisters are not mutants, but rather dormant cells that can survive antimicrobial treatments that kill the majority of their genetically identical siblings. Persister cells have entered a physiological state in which they grow slowly or very slowly that makes them insensitive (refractory or resistant) to the action of antimicrobial drugs. Similarly, "antibiotic resistance" refers to the ability of bacteria to naturally resist killing by antibiotics, and "antifungal resistance" refers to the ability of fungi to naturally resist killing by antibiotics.

[0088] "Antibacterial resistance" refers to the ability of a microorganism to resist the effects of drugs that once successfully treated the microorganism. Microorganisms that are resistant to multiple antibacterial drugs are called multidrug resistant (MDR). Resistance occurs through one of three mechanisms: natural resistance in a particular type of bacteria, genetic mutations, or one species acquires resistance from another. Mutations can lead to inactivation of the drug, changes in the binding site of the drug, changes in metabolic pathways, and reduced permeability of the drug.

[0089] As used herein, the term "in combination" or "in further combination" or "further in combination" refers to the use of additional prophylactic and / or therapeutic agents as well as the BT compositions of the present disclosure. The use of the term "in combination" does not limit the order in which prophylactic and / or therapeutic agents are administered to a subject. A first prophylactic or therapeutic agent can be administered prior to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), concomitantly with, or subsequent to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) administration of a second prophylactic or therapeutic agent (different from the first prophylactic or therapeutic agent) to a subject.

[0090] As used herein, the terms "prophylactic agent" and "prophylactic agents" refer to an agent, such as the BT composition of the present disclosure, that can be used to prevent, manage, or control one or more signs and symptoms of a disease or disorder, particularly a disease or disorder associated with a microbial (e.g., bacterial and / or fungal) infection, such as a diabetic foot infection.

[0091] As used herein, the terms "antibacterial activity", "antifungal activity" and "antimicrobial activity" in relation to the BT compositions of the present disclosure refer to the ability to kill and / or inhibit the growth or replication of a particular microorganism. In certain embodiments, antibacterial or antimicrobial activity is assessed by culturing bacteria, such as gram-positive bacteria (e.g., S. aureus), gram-negative bacteria (e.g., A. baumannii, E. coli, and / or P. aeruginosa), or bacteria not classified as either gram-positive or gram-negative, or fungi according to standard techniques (e.g., in liquid culture or on agar plates), contacting the culture with a BT composition of the present disclosure, and monitoring cell growth after said contact. For example, in liquid culture, bacteria may be grown to an optical density ("OD") that represents the midpoint of the exponential growth of the culture; the culture is exposed to one or more concentrations of one or more BT compounds or variants thereof of the present disclosure, and the OD is monitored relative to a control culture. A reduced OD compared to the control culture represents antibacterial activity (e.g., indicates lytic killing activity). Similarly, bacterial colonies can be formed on agar plates, exposed to the BT composition of the present disclosure or its variants, and the subsequent growth of the colonies can be evaluated relative to the control plate. A reduction in colony size, or a reduction in total number of colonies, indicates antibacterial activity.

[0092] "Biofilm" refers to a symbiotic community of microorganisms in which cells adhere closely to each other and often to surfaces. These attached cells become embedded in a slimy extracellular matrix composed of extracellular polymeric substances (EPS). Once a biofilm is formed, the microorganisms become up to 1000 times more resistant to antibiotics than planktonic bacteria. Bacterial aggregates are clusters of laterally aligned cells that can initiate the development of biofilms with more complex and dense 3D structures. In some embodiments, biofilms may include one or more species of bacteria (e.g., Pseudomonas aeruginosa and Staphylococcus aureus) and / or one or more different phyla (e.g., bacteria, viruses, and fungi).

[0093] The term "infectious disease" is used in the broadest sense herein and refers to any infectious disease, such as a viral infection, or caused by a microbial bacterial infection, fungal infection, or parasitic infection (e.g., protozoa, amoeba, or worm). Examples of such infectious diseases can be found in many well-known textbooks, such as, for example, "Medical Microbiology" (Greenwood, D., Slack, R., Peutherer, J., Churchill Livingstone Press, 2002); "Mims' Pathogenesis of Infectious Disease" (Mims, C., Nash, A., Stephen, J., Academic Press, 2000); "Fields" Virology. (Fields, BN, Knipe DM, Howley, PM, Lippincott Williams and Wilkins, 2001); and "The Sanford Guide To Antimicrobial Therapy" 26th Edition, JP Sanford et al. (Antimicrobial Therapy, Inc., 1996), which are incorporated herein by reference. For example, the presence of infection in diabetic foot wounds is defined by clinical signs and symptoms of infection or inflammation, not by culture of microorganisms that are always present. However, even shortly after resolution of clinical signs and symptoms of wound infection, most patients have an underlying ulcer (e.g., diabetic foot ulcer) that requires ongoing treatment to promote complete wound closure. Of note, however, is that many wound specialists believe that in addition to the clinically defined infectious state, there exists a less clinically apparent pathological condition known as "critical colonization." In this condition, wound healing may be delayed or halted due to the asymptomatic presence of high levels of bacteria. This critical colonization, sometimes referred to as high "wound bioburden," is polymicrobial and often associated with biofilm-producing bacteria; it has been shown to interfere with the normal wound healing process, as it induces or prolongs the active inflammatory phase of repair.The bacterial cells that compose such biofilms are difficult to recognize because they often exist in a viable but non-culturable (VBNC) state (Pasquaroli, 2013), but are attached to surfaces and are usually more resistant and resistant to antibiotics and disinfectants than their planktonic counterparts (Costerton, 1999; Nguyen, 2011).The term "infection" therefore contemplates "significant colonization" as well as a clinically defined infectious state.

[0094] The term "wound closure" can encompass healing of a wound in which the sides of the wound rejoin to form a continuous border (e.g., intact skin). In another embodiment, the compositions and methods provided herein promote tissue regeneration. In another embodiment, the compositions and methods provided herein limit scarring of tissues such as glia, tendons, ocular tissue, ligaments, or skin. In some embodiments, "wound closure" refers to complete or substantially complete re-epithelialization. In some embodiments, "wound closure" occurs via secondary intent.

[0095] It should be understood that the term "wound healing" can encompass the process of regeneration through wound closure and induction of a temporal and spatial healing program that includes the processes involved in wound closure. The term "wound healing" can also encompass the processes of granulation, angiogenesis, fibroblast, endothelial and epithelial cell migration, extracellular matrix deposition, re-epithelialization, and remodeling. In some embodiments, "wound healing" refers to a wound that remains closed for a sufficient period of time (e.g., 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 1 month) after initial wound closure. In some embodiments, "wound healing" refers to a wound that remains closed for 2 weeks after initial wound closure.

[0096] It will be understood by the skilled artisan that the term "granulation" can encompass the process of the formation of small red granular bumps on a raw surface (the surface of a wound or ulcer) as a healing agent. Granulation may also include the formation of granulation tissue on a wound. "Granulation tissue" refers to newly grown tissue material at a wound site that forms to heal the wound. The tissue is a perfused fibrous connective tissue that contains a variety of cell types. The tissue will generally grow from the base of the wound and gradually fill the entire wound space.

[0097] It will be understood by the skilled artisan that the term "angiogenesis" can encompass new growth of blood vessels, resulting in improved oxygen and nutrient supply. Similarly, it will be understood by the skilled artisan that the term "angiogenesis" may encompass the vascularization process involving the development of new capillaries. It will also be understood that the term "cell migration" refers to the movement of cells (e.g., fibroblasts, endothelium, epithelium, etc.) to a wound site.

[0098] It should be understood that the term "extracellular matrix deposition" can include secretion by cells of fibrous factors (e.g., collagen, elastin, reticulin), link proteins (e.g., fibronectin, laminin), and space-filling molecules (e.g., glycosaminoglycans). It will be understood by the skilled artisan that the term "type I collagen" can include the most abundant collagen, which forms large, well-organized fibrils with high tensile strength.

[0099] It will be understood by the skilled artisan that the term "re-epithelialization" can encompass the re-formation of epithelium on an exposed surface (eg, a wound).

[0100] The term "remodeling" refers to replacement of granulation tissue and / or devascularization.

[0101] As used herein, "substantially" or "substantially" refers to the complete or nearly complete extent or degree of an action, feature, characteristic, state, structure, item, or result. For example, an object that is "substantially" surrounded means that the object is completely surrounded or nearly completely surrounded. The exact acceptable degree of deviation from absolute completeness may, in some cases, depend on the particular context. Generally speaking, however, the proximity of completion will be such that the object has the same overall result as if absolute and complete completion had been obtained. The use of "substantially" is equally applicable when used in a negative sense to refer to the complete or nearly complete lack of an action, feature, characteristic, state, structure, item, or result. For example, a composition that is "substantially free" of other active agents is completely devoid of the other active agent, or nearly completely devoid of the other active agent to such an extent that the effect is the same as if the other active agent were completely absent. In other words, a composition that is "substantially free" of a component or element or another active agent may still contain such an item, so long as there is no measurable effect of it.

[0102] As used herein, "D90" refers to the 90% value of the particle size (i.e., fine particles). For example, if D90=1 μm, 90% of the particles are smaller than 1 μm. Similarly, "D80" refers to the 80% value of the particle size (i.e., fine particles), "D70" refers to the 70% value of the particle size (i.e., fine particles), "D60" refers to the 60% value of the particle size (i.e., fine particles), "D50" refers to the 50% value of the particle size (i.e., fine particles), "D40" refers to the 40% value of the particle size (i.e., fine particles), "D30" refers to the 30% value of the particle size (i.e., fine particles), "D20" refers to the 20% value of the particle size (i.e., fine particles), and "D10" refers to the 10% value of the particle size (i.e., fine particles).

[0103] As used herein, MBN-101 refers to a composition that includes BisEDT. How to use In some embodiments, the present disclosure provides a method of treating a topical wound comprising administering to a subject in need thereof a therapeutically effective amount of a composition comprising a bismuth thiol compound, where the composition is applied to the infection (e.g., applied to the surface of the infection).

[0104] Local treatment offers the advantage of avoiding systemic adverse effects, providing high concentration at the target site, and allowing the use of drugs that are not available for systemic therapy. In some embodiments, mechanical debridement may be used to improve local treatment, as it reduces the existing bacterial bioburden and opens the time-dependent therapeutic window of local antimicrobial therapy (TAT) (Wolcott RD, et al. 2010. J. Wound Care, 19: 320-328). Nevertheless, to date, TAT agents have not proven effective in treating DFI (Nelson EA, et al. 2006, Diabet. Med. 23: 348-359).

[0105] In some embodiments, the local wound is a skin ulcer (e.g., a skin ulcer of the lower extremities). In some embodiments, the local wound is a skin ulcer of the lower extremities, such as the leg and / or foot. In some embodiments, the skin ulcer is one or more of a foot ulcer, an ischemic ulcer, a gangrenous ulcer, a venous stasis ulcer, a decubitus ulcer, a Buruli ulcer, or a traumatic ulcer. In some embodiments, the skin ulcer is a foot ulcer. In some embodiments, the local wound is infected with one or more bacterial and / or fungal pathogens. In some embodiments, the local wound is infected with a bacterial pathogen. In some embodiments, the local wound is a diabetic foot ulcer. In some embodiments, the diabetic foot ulcer is a diabetic foot infection.In some embodiments, the local wound is free of the following bacterial pathogens: Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp., Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, Coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium acnes, S.dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas Sp.

[0106] Biofilms of S. aureus and other bacteria present in wounds of patients with DFI increase the difficulty of successful infection management and mitigation. Combinations of DFI-associated bacteria forming multi-species biofilms containing S. aureus have shown greater resistance, virulence, and pathogenicity than comparable single-species biofilms. The presence of such complex biofilms in wounds of patients with DFI is considered to be a major contributor to the chronic and persistent nature of these infections.

[0107] In some embodiments, the bacterial pathogen exhibits resistance to one or more antibiotics. Of particular concern are Methicillin-resistant Staphylococcus aureus strains (MRSA). MRSA was a rare occurrence in the hospital environment until the 1990s, when the prevalence of MRSA in hospitals exploded. MRSA is now considered endemic to hospitals, especially in the UK (Johnson AP et al, 2001, J. Antimicrobial Chemotherapy, 48(1):143-144). Furthermore, MRSA presents a new threat in diabetic foot infections (Retrieved January 17, 2009, from the CDC: Centers for Disease Control and Prevention website). Ulcers and open wounds that can develop in diabetic feet put patients at risk of infection with MRSA, and recent studies have shown evidence that MRSA impairs healing when present in diabetic wounds (Bowling FL, et al. 2009, Curr Diab Rep 9(6):440-444). See also Kosinski, MA, et al. 2010. Expert Rev AntiInfect Ther. 8(11):1293-1305. In some embodiments, bacterial pathogens are treated with amikacin, methicillin, vancomycin, nafcillin, gentamicin, metronidazole, piperacillin / tazobactam, ampicillin, chloramphenicol, doxycycline, tobramycin, levofloxacin, cephalosporins (e.g., cephalexin, cefoxitin, ceftizoxime, ceftibiprole, ceftazidime, ceftaroline), penicillin / β-lactamase inhibitor combinations (e.g., cephalexin, cefoxitin, ceftizoxime, ceft ... For example, the infection is resistant to known standard antibiotic treatments, including, but not limited to, amoxicillin / clavulanate, ampicillin / sulbactam, piperacillin / tazobactam, and ticarcillin / clavulanate), carbapenems (e.g., imipenem / cilastatin, ertapenem), fluoroquinolones (e.g., ciprofloxacin, moxifloxacin), clindamycin, linezolid, daptomycin, tigecycline, and vancomycin.

[0108] Long-term repeated treatment with antibiotics to treat DFI-associated infections may result in the development of antibiotic resistance, characterized by the presence of microbial biofilms. Recent studies have repeatedly demonstrated a correlation between multidrug-resistant (MDR) bacteria and their ability to form stronger and more productive biofilms. Bacteria within biofilms are protected from antibiotics, enhancing the minimum inhibitory concentration of such antibiotics.

[0109] The BT compositions of the present disclosure are active against multiple bacterial and fungal strains. In some embodiments, the BT composition is selected from the group consisting of Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp, Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, Coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium It has activity against multiple strains including, but not limited to, Acne, S. dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter clacae, Sphingomonas sp., Acinetobacter spp., Anaerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, and Anaerococcus vaginalis.Thus, some embodiments of the present disclosure provide for the prevention and treatment of Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp, Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, Coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium spp., and Propionibacterium spp. in humans and animals using BT compositions. In another aspect, the disclosure provides a method for treating and / or preventing infections associated with Acinetobacter pneumoniae, S. dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anaerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis. In another aspect, the disclosure provides a method for treating and / or preventing infections associated with related species or strains of these bacteria.In some embodiments, the bacterial infection is an infection associated with a diabetic leg infection, such as a diabetic foot infection.

[0110] Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp, Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium acnes, S.dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anaerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, and Anaerococcus vaginalis are involved in a number of severe opportunistic infections, especially in immunocompromised patients, including those with diabetes.The pharmaceutical composition of the present disclosure can be applied to Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp, Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium acnes, S.dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis It is contemplated to treat and / or prevent infections associated with, or other species or strains of bacteria, including, but not limited to, infections of the skin, infections in and around wounds, chronic ulcers, ulcers associated with burns, post-operative infections, infections associated with catheters and surgical drains, and blood infections.In some embodiments, the pharmaceutical compositions of the present disclosure are used in the treatment and / or prevention of bacterial infections associated with areas of broken skin, including, but not limited to, infections associated with skin ulcers such as diabetic foot ulcers, skin lesions, vesicles, cysts, blisters, blisters, open wounds such as decubitus ulcers (bedsores) and other pressure sores, chronic ulcers, cellulitis and associated sores, erysipelas and associated lesions, wounds, burns and associated wounds, pimples, or other conditions where the skin is damaged, cracked, broken, torn, and otherwise damaged.

[0111] In any of the embodiments described herein, the BT composition may be used to treat infections (e.g., DFIs) with one or more of the following bacterial pathogens: Acinetobacter baumanii Acinetobacter junii Anaerococcus lactolyticus Anaerococcus vaginalis Anaerococcus murdoch Anaerococcus tetradius Anaerococcus hydrogenalis Actinobaculum massiliense Actinobaculum schaalii Actinomyces europaeus Actinomyces hominis Actinomyces neuii Actinomyces radingae Alcaligenes faecalis Abiotrophia paraadiacens Bacteroides fragilis Bulleidia extructa Bilophila wadsworthia Campylobacter ureolyticus Citrobacter murliniae Clostridium saccharogum Clostridium novyi Corynebacterium accolens Corynebacterium amycolatum Corynebacterium aurimucosum Corynebacterium freiburgense Corynebacterium hansenii Corynebacterium jeikeium Corynebacterium mycetoide Corynebacterium simulans Corynebacterium Tuberculostearicum Corynebacterium xerosis Corynebacterium striatum Dermabacter of man The unpopular dialer Dialister propionificaciens Dialister microaerophilus Dialister pneumosints Delftia acidivorous Enterobacter aerogenes Enterobacter cloacae Enterobacter hormaechei Enterococcus faecalis Enterococcus canintestinum Escherichia coli Escherichia fergusonii Escherichia coli Enterococcus avium Enterococcus fowleri Enterococcus casseliflavus Eggerthella lenta Finegoldia magna Fusobacterium canifelinum Fusobacterium nucleatum Fusobacterium periodontium Granulicatella adiacens The twin of measles Blood clot Haemophilus parainfluenzae Haemophilus indolentis Helicobacter kunzii Helicobacter kunzii Klebsiella oxytoca Black Kocuria Leclercia adecarboxylata Mobiluncus curtisii Morel indoligenes Morganella morganii Negativicocus succinicivorans Peptoniphilus harei Peptoniphilus gorbachii Ivory-billed Peptoniphilus Peptoniphilus lacrimalis Peptoniphilus olsenii Peptoniphilus asacchrolyticus Parvimonas micra Peptococcus niger Peptostreptococcus anaerobic Peptostreptococcus stomatis Porphyromonas asaccharolytica Porphyromonas bennonis Porphyromonas somerae Porphyromonas veononis Porphyromonas levii Prevotella timonensis Prevotella bergensis Oral thrush Body hair Prevotella dissident Prevotella intermedia Prevotella nanceiensis Pseudomonas indica Pseudomonas otitis Psychrobacter lutiphocae Proteus mixofacias Proteus hauseri Providence Rettgeri Providence of Stuart Staphylococcus aureus Staphylococcus epidermidis Staphylococcus carnivorous Staphylococcus chromogenes Staphylococcus devriesei Staphylococcus aureus Staphylococcus lugdunensis Serratia nematodiphila Stenotrophomonas maltophilia Staphylococcus pettenkoferi Staphylococcus aureus Staphylococcus saprophyticus Streptococcus agalactiae Streptococcus anginosus Streptococcus canis Streptococcus dysgalactiae Streptococcus infantis Oral Streptococcus Serratia grimesii Stenotrophomonas pavanii Salmonella enterica Trueperella bernardiae Varibaculum cambriense Veillonella atypical Veillonella small Veillonella dispar Veillonella rogosae Acinetobacter calcoaceticus Acinetobacter lwoffii Anaerococcus prevotii Bacteroides faeces Bacteroides distasonis Bacteroides ovatus Bacteroides suffocatus Bacteroides thetaiotaomicron Bacteroides uniformis Bacteroides vulgaris Corynebacterium striatum Harmless Clostridium Clostridium perfringens Clostridium ramosum Pluralibacter gergoviae Fusobacterium deadly Finegoldia magna Klebsiella oxytoca Klebsiella pneumoniae Pseudomonas aeruginosa Peptococcus magnus Prevotella bivia Prevotella melaninogenica Porphyromonas asaccharolytica Peptostreptococcus asaccharolyticus Peptostreptococcus micros Parvimonas micra Miraculous Proteus Staphylococcus haemolyticus Staphylococcus simulans Staphylococcus saprophyticus Streptococcus pneumoniae Streptococcus agalactiae Streptococcus milius Streptococcus milleri Streptococcus dysgalactiae Streptococcus canis Serratia marcescens Serratia liquefaciens Stenotrophomonas maltophila Epidermolysis bullosa In any of the embodiments described herein, the BT composition may be used to treat an infection (e.g., DFI) with one or more of the following fungal pathogens: Candida spp., Cladosporium spp., Aspergillus spp., Penicillium spp., Alternaria spp., Pleospora spp., Fusarium spp, Candida lusitaniae, Candida parapsilisis, and Candida albicans.

[0112] In some embodiments, the BT composition of the present disclosure is used to treat chronic ulcers. Chronic ulcers may develop from wounds caused by various factors, especially in patients with impaired blood circulation caused by, for example, cardiovascular problems or external pressure from a bed or wheelchair. More than 8 million patients are diagnosed with chronic skin ulcers every year in the United States alone (Harsha, A. et al., 2008, Journal of Molecular Medicine, 86(8):961-969), which costs more than $10 billion annually (Margolis, DJ, et al., 2002, Journal of the American Academy of Dermatology, 46(3):381-386). Chronic ulcers may develop in the mouth, throat, stomach, and skin. Chronic skin ulcers include diabetic ulcers, venous ulcers, radiation ulcers, and pressure ulcers, and the three major categories of chronic skin ulcers are diabetic ulcers, venous stasis ulcers, and pressure ulcers. Chronic ulcers cause significant loss of integrity of the skin and can lead to morbidity and even mortality.

[0113] In some embodiments, the BT composition of the present disclosure is used to treat diabetic foot infections, such as diabetic foot infections. Diabetic foot infections are one of the major complications of diabetes mellitus, occurring in about 15% of all diabetic patients and accounting for about 85% of all lower limb amputations (Brem, et al., J. Clinical Invest., 2007, 117(5):1219-1222). Because diabetes mellitus interferes with the normal steps of the wound healing process, diabetic foot infections may become associated with chronic skin ulcers that do not heal.

[0114] Chronic wounds represent a failure of the normal process of acute wound healing. Wound healing has traditionally been divided into three distinct phases: inflammation, proliferation, and remodeling. The inflammation phase of wound healing begins at the time of injury by forming a clot via a platelet plug, thereby initiating a response from neutrophils and macrophages. Neutrophils first clear the damage of bacteria and debris by releasing various proteases and reactive oxygen free radicals. Macrophages are then attracted to the wound site by chemotactic factors, which subsequently release their own chemotactic factors to stimulate fibroblasts and more macrophages. During the proliferation phase, fibroblasts initiate epithelialization, angiogenesis, and collagen formation. Epithelialization generally occurs from the basement membrane if it remains intact, or from the wound edge if it is not. Fibroblasts synthesize type III collagen during this phase and turn into myofibroblasts, which help stimulate wound contraction. During the remodeling phase, type III collagen begins to replace type I collagen. The collagen is woven into an organized cross-linked network, the strength of which approaches 80% of that of the original, intact tissue.

[0115] There are many factors that can stall the three-phase healing process and transform acute wounds into chronic wounds. These may include low proliferation capacity of fibroblasts, downregulation of receptors, reduction of growth factors, or lack of suitable protein matrix in the dermis. In addition, insufficient perfusion and / or nutrition can cause wounds to stall at the inflammatory stage and excessive accumulation of exudate in the wound. Chronic ulcers can be considered as non-healing areas of wounded skin, such as areas of wounded skin that fail to follow the normal process of wound healing and / or fail to respond or respond appropriately to initial treatment, as described above. Chronic ulcers of the skin may be characterized as wound lesions that last for more than 4 weeks without significant healing tendency, or as wounds that frequently recur (Fonder, M. et al., 2012, Journal of the American Academy of Dermatology, 58(2):185-206). Chronic wounds may appear with red granulation tissue and yellow pus, dusky purple skin around the granulation tissue, or gray-white swollen granulation tissue. Standard treatment procedures for chronic skin ulcers include, for example: removal of necrotic or infected tissue; establishment of adequate blood circulation; maintenance of a moist wound environment; management of wound infection; wound cleansing; and nutritional support, including blood sugar control, in subjects with diabetic ulcers. For example, in diabetic patients, poor control of blood sugar levels allows bacteria to grow even more rapidly in the wound; furthermore, neurodegeneration in diabetes means that the condition may not be painful and therefore may go undetected, at least initially. Chronic ulcers, including diabetic foot ulcers, often become further infected with opportunistic bacteria, leading to a worsening of the condition.Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp, Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium acnes, S. dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis are associated with such infections.

[0116] In some embodiments, the pharmaceutical compositions of the present disclosure are selected from the group consisting of Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp, Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, Coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium The present invention is formulated for use in a method of treating and / or preventing bacterial infections caused by Acinetobacter spp., S. acnes, S. dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anaerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis.In some embodiments, the pharmaceutical compositions of the present disclosure are selected from the group consisting of Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp, Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, Coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium The present invention is formulated for use in a method of treating and / or preventing bacterial infections caused by Acinetobacter spp., S. acnes, S. dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anaerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis.In some other embodiments, the pharmaceutical compositions of the present disclosure are selected from the group consisting of Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp, Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium The composition is formulated for use in a method of treating and / or preventing a bacterial infection caused by bacteria other than Acinetobacter acnes, S. dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anaerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis.

[0117] In some embodiments, the present disclosure provides a method of treating and / or preventing a chronic ulcer comprising administering to a subject in need thereof a therapeutically or prophylactically effective amount of a pharmaceutical composition of the present disclosure. In some embodiments, the administration comprises topical administration to an area of ​​damaged skin associated with the chronic ulcer. In some embodiments, the topical administration is followed by debridement of the area to be treated.

[0118] In some embodiments, the present disclosure provides a method of treating and / or preventing a diabetic foot infection comprising administering to a subject in need thereof a therapeutically or prophylactically effective amount of a pharmaceutical composition of the present disclosure. In some embodiments, the administration comprises topical administration to an area of ​​broken skin associated with a diabetic foot infection, e.g., a skin ulcer. In some embodiments, the topical administration is followed by debridement of the area to be treated.

[0119] Debridement can be accomplished by a number of approaches. Surgical debridement involves excising dead tissue from the wound or other areas of injured skin. Mechanical debridement uses a variety of methods, such as pressure irrigation devices, whirlpool water baths, ultrasound, larval maggots, or specialized bandages to loosen and remove wound debris. Autolytic debridement augments the body's natural process of mobilizing enzymes that break down dead tissue, for example, by using appropriate bandages that keep the wound moist and clean. Enzymatic debridement uses chemical enzymes and appropriate bandages to further help destroy dead tissue at the site of the wound or other areas of injured skin.

[0120] Debridement improves local treatment because it reduces the bacterial bioburden present and opens a time-dependent therapeutic window for topical antimicrobial therapy (TAT) (Wolcott RD, et al. 2010. J Wound Care 19:320-328). With regard to the timing of debridement, early or immediate debridement is preferred over delayed debridement once this treatment option has been selected in wound management. Furthermore, multiple debridements during wound management may be indicated (Wolcott RD, et al. 2009. J Wound Care 18(2):54-6). For example, in some embodiments, debridement precedes the topical application of the BT composition of the present disclosure and is repeated before every administration of the BT composition. In some embodiments, debridement is performed only before every other administration of the BT composition, or only before every third, fourth, fifth, or sixth administration of the BT composition. In some embodiments, debridement is performed less frequently than the application of the BT composition, for example, once a week. Therefore, if the BT composition is applied daily, the patient will not be debridement every time it is applied. In some embodiments, whether or not the wound is debridement before the topical administration of the BT composition of the present disclosure is within the clinical judgment of the medical practitioner treating the wound, such as a physician, physician assistant, or emergency medical practitioner.

[0121] The BT compositions of the present disclosure can be used to treat, manage, control, and / or prevent infections associated with chronic ulcers, including diabetic foot infections and associated skin ulcers. In other embodiments, the BT compositions of the present disclosure are used to treat, manage, control, and / or prevent microbial (e.g., bacterial and / or fungal) infections associated with other areas of injured skin, such as cellulitis sores, erysipelas lesions, decubitus ulcers, burns, traumatic wounds, and pressure sores. In some such embodiments, the compositions used may be topical compositions formulated for local administration, e.g., for direct application to areas of injured skin, as described above.

[0122] The BT composition of the present disclosure can also be used to treat, manage, control, and / or prevent decubitus ulcers.Decubitus ulcers, also known as bed sores or pressure sores, are damage to the skin and underlying tissue caused by prolonged pressure on the area.For example, pressure sores most often occur on the skin covering bony parts of the body, such as the heel, ankle, hip, and buttocks.

[0123] Pressure ulcers are classified into one of four stages based on their severity. Stage I is the early stage of a pressure ulcer while the skin is still intact. The skin may appear red, gray, bluish, or purple and does not blanch to the touch. Stage II often involves an open wound of broken skin. At this stage, the outer layer of skin (epidermis) and part of the skin layer underneath (dermis) are damaged or lost. The ulcer may appear as a shallow pinkish-red basin-shaped wound. In stage III, the ulcer is a deep wound and some fat may be exposed due to loss of skin, and the ulcer has a crater-like appearance. The wound base may also have yellowish dead tissue (scab). Stage IV ulcers show extensive loss of tissue and the wound may expose muscle, bone, and tendons. The wound base may contain a scab, or dark, crusty dead tissue (eschar).

[0124] Similar to the treatment of diabetic foot ulcers, debridement (e.g., of bedsores or pressure sores) can be used to remove damaged, dead, or infected tissue from the wound and promote proper healing, e.g., as described herein and / or otherwise known in the art. In some embodiments, administration of the pharmaceutical composition of the present disclosure follows debridement. For example, the pharmaceutical composition disclosed herein may be administered topically to a bedsore ulcer following its surgical, mechanical, autolytic, or enzymatic debridement.

[0125] The BT composition of the present disclosure is also used for the treatment, management, control, and / or prevention of cellulitis and / or erysipelas, including but not limited to the sores and lesions associated with cellulitis and erysipelas. Cellulitis and erysipelas are skin infections that develop as a result of bacterial invasion through a breach in the skin's protective barrier. For example, cracks in the skin, cuts, blisters, burns, insect bites, spider bites, tattoos, surgical wounds, intravenous drug injections, or intravenous catheter insertion sites may provide an avenue of bacterial entry. Group A Streptococcus and Staphylococcus are the most common bacteria involved in cellulitis. Cellulitis is most frequently observed in middle-aged and elderly people, while erysipelas occurs in children and the elderly (Ellis Simonsen SM et al. 2006. Epidemiol Infect. 134(2):293; and Eriksson B. et al. 1996, Clin Infect Dis 23:1091). Also at increased risk are individuals with immunodeficiencies, diabetes, alcoholism, fungal infections, and impaired lymphatic drainage. Diabetics are particularly prone to cellulitis of the feet because the disease causes impaired blood circulation in the feet. The lower legs are the most common site of infection for both erysipelas and cellulitis (Ellis Simonsen SM et al. 2006. Epidemiol Infect. 134(2):293; Chartier C et al 1996 Int J Dermatol 35:779).

[0126] Cellulitis and erysipelas often coexist and commonly manifest as areas of erythema, edema, and warmth of the skin. They differ in that erysipelas involves the upper dermis and superficial lymphatics, whereas cellulitis involves the deep dermis and subcutaneous fat. Thus, erysipelas has more distinctive anatomical features than cellulitis—erysipelas lesions may be elevated above the level of the surrounding skin, with a clear demarcation between involved and uninvolved tissue (Bisno AL et al.1996 N Engl J Med 334:240). Lesions may appear as red, swollen, warm, indurated, and / or a rash consistent with and resembling the peel of an orange. Erysipelas may appear on the face, for example, in a "butterfly" pattern. More severe infections can produce blisters, bullae, and petechiae with the potential for skin necrosis. In addition, patients with erysipelas tend to develop acute signs and symptoms with systemic symptoms, including fever and chills.

[0127] Patients with cellulitis tend to have a more gradual course of progression, with signs and symptoms appearing over a period of days. Various forms of cellulitis include periorbital cellulitis, abdominal wall cellulitis (in morbidly obese individuals), oral cellulitis (due to Streptococcus pneumoniae), Ludwig's angina (submandibular space cellulitis), and perianal cellulitis (due to group A beta-hemolytic streptococci) (Barzilai A,et al,1998 Pediatr Infect Dis J.17(4):358;Thorsteinsdottir B,et al.2005 Scand J Infect Dis.37(8):605). Cellulitis can also cause flu-like signs and symptoms with high fever and shivering.

[0128] In some embodiments, the treatment of cellulitis or erysipelas further comprises the administration of an antibiotic. For example, the pharmaceutical composition of the present disclosure may be administered locally to the erysipelas lesion in combination with an antibiotic selected from the group consisting of penicillin, clindamycin, and erythromycin. As another example, the pharmaceutical composition of the present disclosure may be administered locally to the pain associated with cellulitis in combination with an antibiotic selected from the group consisting of flucloxacillin, dicloxacillin, penicillin, ampicillin, and amoxicillin. The antibiotic may be administered orally, intravenously, or topically, for example, in conjunction with the topical administration of the BT composition of the present disclosure.

[0129] The BT composition of the present disclosure is also used to treat, manage, control, and / or prevent infections associated with burns. A burn is an area of ​​damaged skin caused directly or indirectly from a burn. A burn is a type of skin injury that can be caused by heat, as well as electricity, chemicals, light, radiation, and friction. A burn may only affect the skin (epidermal tissue), but in some cases may also damage deeper tissues such as muscles, bones, and blood vessels. Burns can be classified by the mechanism of injury, depth, extent, associated injuries, and comorbidities. Burns are traditionally described based on the depth of dermal damage and are broadly classified as first-, second-, third-, or fourth-degree burns (Walls et al., 2009, Rosen's Emergency Medicine: Expert Consult Premium Edition (Rosen's Emergency Medicine: Concepts & Clinical Practice (2v)). Important characteristics of a burn include its cause (thermal, chemical, electrical), anatomical location, depth (total or partial thickness), duration, and extent (percentage of total body surface area). Patient characteristics that affect burn healing include age, nutritional status, underlying medical conditions, and concomitant injuries (e.g., head trauma, inhalation injury, fractures).

[0130] Infections in burn patients are a major problem, with reported incidence rates of nosocomial infections varying from 63 to 240 per 100 patient-days and 53 to 93 per 1000 patient-days, depending primarily on the definition used (Chim H, et al, 2007, Burns 33:1008-1014; and Wibbenmeyer L, et al., 2006, J Burn Care Res 27:152-60). Furthermore, bacterial infections of burns are associated with adverse outcomes and mortality. In a series of 175 patients with severe burns, for example, infection preceded multiple organ failure in 83% of patients and was considered the direct cause of death in 36% of patients who did not survive (Fitzwater J, et al., 2003, J Trauma 54:959-66). Burns may become infected from multiple sources. Burns may be initially infected with gram-positive bacteria, primarily staphylococci, which are normal deep residents of sweat glands and hair follicles exposed by burns (Sharma B R., 2007, Infect Dis Clin North Am 21:745-59; ix). Moist vascular burn eschar may further support microbial growth. Gram-negative infections may result from metastasis from the colon, for example, due to reduced mesenteric blood flow at the time of the burn and subsequent injury (Herndon DN, et al., 2000, Crit Care Med 28:1682-3). Additionally, burn patients may develop immune disorders including impaired cytotoxic T lymphocyte responses, bone marrow maturation arrest leading to neutropenia, impaired neutrophil function, and reduced macrophage production (Sharma B R., 2007, Infect Dis Clin North Am 21:745-59, ix; Gamelli RL, et al., 2000, J Burn Care Rehabil 21:64-9; Hunt JP, et al., 1998, J Surg Res 80:243-51; and Shoup M, et al., 1998, Ann Surg 228:112-22).Finally, burn patients are susceptible to hospital-acquired infections common to other intensive care patients, including intravascular catheter-associated infections and ventilator-associated pneumonia, and the overall incidence of infections is higher than that of other intensive care patients (Chim H, et al., 2007, Burns 33:1008-14; and Wibbenmeyer L, et al., 2006, J Burn Care Res. 27:152-60). In fact, most manifestations of bloodstream infections in burn patients after the first week are caused by hospital-acquired multidrug-resistant bacteria (Wibbenmeyer L, et al., 2006, J Burn Care Res 27:152-60; and Ressner RA, et al., 2008, J Am Coll Surg 206:439-44).

[0131] Conventional treatment of burns includes debridement and excision, application of wound dressings, wound closure, skin grafting, fluid resuscitation, management of wound infection such as administration of antibiotics, pain management, nutritional support, and / or measures to inhibit excessive scar formation. Burns may be covered with clean, dry sheets or dressings (e.g., plastic wrap). Early cooling with cold water within 30 minutes of the burn reduces burn depth and pain. Debridement, cleaning, and dressing are important aspects of burn care.

[0132] In some embodiments, the treatment of the burn further comprises the administration of an antibiotic. It has been shown that antibiotic prophylaxis may reduce mortality, bacteremia, and ventilator-associated pneumonia among patients in intensive care units (Silvestri L, et al, 2007, J. Hosp. Infect. 65:187-203; and De Smet AM, et al., 2009, N. Engl. J. Med. 360:20-31). In burn patients, the skin is an additional source of infection (Avni T, et al., 2010, BMJ, 340:c241). In some embodiments, the treatment of the burn further comprises the administration of an agent for pain management. The pharmaceutical composition of the present disclosure may be administered topically to the burn in combination with an agent for pain management selected from the group consisting of simple analgesics, ibuprofen, acetaminophen, and anesthetics. The antibiotic and / or agent for pain management may be administered orally, intravenously, or topically, for example, in conjunction with the topical administration of the BT composition of the present disclosure. One or more other aspects of conventional treatment for burns may also be used in combination with the BT compositions of the present disclosure.

[0133] In some embodiments, pain management medications for use in combination with the BT compositions of the present disclosure include one or more medications selected from the group consisting of paracetamol (acetaminophen), nonsteroidal anti-inflammatory drugs, ibuprofen, ketoprofen, piroxicam, hydrocodone, morphine, hydromorphine, oxymorphone, fentanyl, oxycodone, diamorphine, methadone, buprenorphine, meperidine, pentazocine, dextromoramide, dipipanone, amitriptyline, dilaudid, tapentadol, and methadone. Pain management medications may include any other medications for pain described herein and / or known in the art.

[0134] In some embodiments, the pain management agent is one that can be applied topically, such as a local anesthetic. A local anesthetic is a local anesthetic used to numb the surface of a body part, such as any area of ​​the skin, the front of the eye, the inside of the nose, ear or throat, the anus, or the genital area. In some embodiments, the pain management agent for use in combination with the BT composition of the present disclosure includes one or more local anesthetics selected from the group consisting of benzocaine, butamben, dibucaine, lidocaine, oxybuprocaine, pramoxine, prilocaine, proparacaine, proxymetacaine, and tetracaine (ametocaine). Local anesthetics are available in creams, ointments, aerosols, sprays, lotions, and jellies. In further embodiments, the local anesthetic may be used with one or more additional agents for pain management, such as another local anesthetic, or a different agent for pain management, such as any other agent(s) for pain management described herein and / or known in the art.

[0135] The BT composition of the present disclosure will include a therapeutically and / or prophylactically effective amount of one or more BT compounds (e.g., BisEDT) as described herein. Therapeutically and / or prophylactically effective amounts refer to the amount necessary to provide a therapeutic and / or prophylactic benefit, respectively, in the subject to which said amount is administered. The therapeutically and / or prophylactically effective amount will depend on the particular formulation, route of administration, condition being treated, whether other drugs or therapies are used in combination with the method of the present disclosure, and other factors.

[0136] In some embodiments of the methods of treating a topical wound, the subject experiences one or more of the following results after completion of dosing: the wound is healed or substantially healed within 12 weeks of the initial administration of the composition; and / or Preventing amputations and / or infection-related surgeries; and / or The wound closes partially or completely; and / or the wound is reduced in size from about 1% reduction compared to the original wound size to complete elimination of the wound; and / or The wound is more than 30 days old and is healed or substantially healed. In some embodiments, the subject experiences two or more of the recited results. In some embodiments, the subject experiences three or more of the recited results. In some embodiments, the subject experiences four or more of the recited results. In some embodiments, the subject experiences all of the recited results. In some embodiments of the methods of treating a localized wound, the subject experiences one or more of the following results after completion of dosing: reduction in reinfection / recurrence 12 weeks after initiation of treatment; elimination or improvement of signs and / or symptoms of infection including redness, swelling, induration, exudate, pain, warmth (at the site of infection) or fever; improved quality of life; eradication of damaging pathogens and / or biofilms; reduced need for concurrent systemic antibiotics.

[0137] In some embodiments, administration of a therapeutically effective amount of a BT composition according to the present disclosure results in improved wound closure (partial or complete), e.g., a reduction in the area of ​​damaged skin (wound area) compared to the area before treatment begins. Wound area can be expressed as a percentage of the initial wound area at one or more time points after treatment begins. For example, in some embodiments, wound area is reduced by at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%; or at least about 90% over the course of treatment with the BT composition of the present disclosure. In some specific embodiments, the reduction in wound area occurs by at least 1 day after the start of treatment (t1), 2 days after the start of treatment (t2), 3 days after the start of treatment (t3), 4 days after the start of treatment (t4), 5 days after the start of treatment (t5), 6 days after the start of treatment (t6), 7 days after the start of treatment (t7), 8 days after the start of treatment (t8), 9 days after the start of treatment (t9), 10 days after the start of treatment (t10), 12 days after the start of treatment (t12), 15 days after the start of treatment (t15), 20 days after the start of treatment (t20), 25 days after the start of treatment (t25), or 30 days after the start of treatment (t30).

[0138] In some embodiments, administration of a therapeutically effective amount of the BT composition according to the present disclosure results in improved wound healing, for example, an improvement in ulcer grade based on the PEDIS classification, compared to the ulcer grade before treatment began. PEDIS is a routinely used and validated classification system for wound-related infections developed by the International Working Group on Diabetic Foot (IWGDF). The IWGDF specifically developed a system for classifying wounds associated with diabetic foot infections using the acronym PEDIS, which stands for perfusion, extent (size), depth (tissue loss), infection, and sensation (neuropathy). The classification was originally developed as a research tool (Schaper N C., 2004, Diabetes Metab Res Rev 20(Suppl 1):S90-5), and provides a semi-quantitative grading of the severity of each category. Specifically, PEDIS grade 1 corresponds to the absence of symptoms or signs of infection; grade 2 corresponds to a localized infection involving only the skin and subcutaneous tissues (no deeper tissue involvement and no systemic signs) while the erythema involved must be between 0.5 cm and 2 cm; grade 3 corresponds to a localized infection as described in grade 2, but involving erythema >2 cm or involving structures deeper than the skin and subcutaneous tissues (e.g., abscess, osteomyelitis, septic arthritis, fasciitis) but without signs of a systemic inflammatory response; and grade 4 corresponds to a localized infection as described in grades 2 and 3, but involving the following: body temperature >38°C or <36°C; heart rate >90 beats / min; respiratory rate >20 breaths / min, or arterial carbon dioxide pressure <32 mmHg. Hg; and a leukocyte count >12000 or <4000 cells / μL or >10% immature (banded) morphology corresponds to a localized infection with signs of systemic inflammatory response syndrome (see, e.g., Lipsky, BA, et al., 2012, CID 54:e132-e173). Another classification system has been developed by the IDSA (Infectious Diseases Society of America) that grades the severity of infection in infected wounds, particularly diabetic foot infections.Specifically, the IDSA grades PEDIS grades 1-4 as "not infected," "mild," "moderate," and "severe," respectively (see again Lipsky, BA, et al., 2012, CID 54:e132-e173).

[0139] In some embodiments, the PEDIS grade is reduced from grade 4 to grade 3, grade 2, or grade 1 over the course of treatment with the BT composition of the present disclosure. In some embodiments, the PEDIS grade is reduced from grade 3 to grade 2 or grade 1 over the course of treatment with the BT composition of the present disclosure. In some embodiments, the PEDIS grade is reduced from grade 2 to grade 1 over the course of treatment with the BT composition of the present disclosure. In some embodiments, the reduction in ulcer grade occurs at least 1 day after initiation of treatment (t1), 2 days after initiation of treatment (t2), 3 days after initiation of treatment (t3), 4 days after initiation of treatment (t4), 5 days after initiation of treatment (t5), 6 days after initiation of treatment (t6), 7 days after initiation of treatment (t7), 8 days after initiation of treatment (t8), 9 days after initiation of treatment (t9), 10 days after initiation of treatment (t10), 12 days after initiation of treatment (t12), 15 days after initiation of treatment (t15), 20 days after initiation of treatment (t20), 21 days after initiation of treatment (t22), 22 days after initiation of treatment (t23), 23 days after initiation of treatment (t24), 24 days after initiation of treatment (t25), 25 days after initiation of treatment (t26), 26 days after initiation of treatment (t27), 27 days after initiation of treatment (t28), 28 days after initiation of treatment (t29), 29 days after initiation of treatment (t30), 30 days after initiation of treatment (t31), 31 days after initiation of treatment (t32), 32 days after initiation of treatment (t33), 33 days after initiation of treatment (t34), 34 days after initiation of treatment (t35), 35 days after initiation of treatment (t36), 36 days after initiation of treatment (t37), 37 days after initiation of treatment (t38), 38 days after initiation of treatment (t39), 39 days after initiation of treatment (t39), 40 days after initiation of treatment (t40), 40 days after initiation of treatment (t41), 41 days after The reduction in ulcer grade occurs by 25 days after initiation of treatment (t25), 30 days after initiation of treatment (t30), 35 days after initiation of treatment (t35), 40 days after initiation of treatment (t40), 45 days after initiation of treatment (t45), 50 days after initiation of treatment (t50), 55 days after initiation of treatment (t55), 60 days after initiation of treatment (t60), 65 days after initiation of treatment (t65), 70 days after initiation of treatment (t70), 75 days after initiation of treatment (t75), 80 days after initiation of treatment (t80), or 85 days after initiation of treatment (t85+). In some embodiments, the reduction in ulcer grade occurs 85 days after initiation of treatment (t85+).

[0140] In some embodiments of the method for treating a topical wound, the BT composition comprises one or more BT compounds selected from BisBAL, BisEDT, bis-dimercaprol, BisDTT, bis-2-mercaptoethanol, bis-DTE, bis-Pyr, bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, BisPyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, bismuth-1-mercapto-2-propanol, and bis-EDT / 2-hydroxy-1-propanethiol, and the composition is a suspension of microparticles comprising the BT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT compound is BisEDT.

[0141] In some embodiments of the method of treating a topical wound, the BT composition comprises BisEDT, and the BisEDT applied is about 20 μg / cm 2 In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0142] In some embodiments of the method of treating a topical wound, following administration of the BT composition, one or more of the following occurs: (i) reducing and / or dispersing microbial (e.g., bacterial and / or fungal) biofilms, (ii) impairing the growth or formation of microbial (e.g., bacterial and / or fungal) biofilms, and (iii) preventing the reformation or spread of microbial (e.g., bacterial and / or fungal) biofilms. In some embodiments, the BT composition treats, manages, and / or reduces the severity of a diabetic foot infection by one or both of (i) preventing infection by bacterial or fungal pathogens and / or (ii) reducing bacterial or fungal pathogens. In some embodiments, the BT composition treats, manages, or reduces the severity of an infection by one or more of: (i) preventing the production or secretion of exotoxins from bacterial or fungal pathogens; (ii) inhibiting cell viability or cell proliferation of planktonic cells of bacterial or fungal pathogens; (iii) inhibiting biofilm formation by bacterial or fungal pathogens; (iv) inhibiting the invasiveness of biofilms or microbial pathogens into underlying tissues (e.g., subcutaneous tissue); (v) inhibiting the virulence of biofilms or microbial pathogens into underlying tissues (e.g., subcutaneous tissue); (vi) inhibiting biofilm viability or biofilm growth of biofilm-forming cells of bacterial or fungal pathogens; and / or (vii) preventing biofilm reformation after debridement.

[0143] BT compounds are known as broad-spectrum antibacterial (and antibiofilm) small molecule drugs for the treatment of chronic and ultimately life-threatening infections. Their efficacy extends to Gram-positive (aerobic and anaerobic) antibiotic-resistant pathogens, including methicillin-resistant Staphylococcus aureus (MRSA, including community-associated [CA]-MRSA), methicillin-resistant Staphylococcus epidermidis (MRSE), and vancomycin-resistant Enterococcus (VRE). BT compounds are also potent against multidrug-resistant (MDR) Gram-negative pathogens (aerobic and aerobic), including Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae (including carbapenem-resistant strains of all the aforementioned bacteria), and Acinetobacter baumannii.

[0144] BT compounds have (a) a highly diverse spectrum of antibiotic resistance profiles (due to evolution / diversification caused by persistence, time, and isolation in many different anatomical regions) and (b) a dual ability to overcome antibiotic resistance and MDR biofilms.

[0145] In some embodiments, the infection contains one or more bacterial or fungal pathogens. In some embodiments, the disclosed methods include treating a DFI-associated infection. In some embodiments, the disclosed methods include managing a DFI-associated infection. In some embodiments, the disclosed methods include reducing the severity of a DFI-associated infection.

[0146] In some embodiments, the bismuth thiol composition comprises a plurality of microparticles comprising a bismuth thiol (BT) compound, substantially all of the microparticles having a volume mean diameter of about 0.4 μm to about 5 μm, wherein the BT compound comprises bismuth or a bismuth salt and a thiol-containing compound. In some embodiments, the bismuth salt is bismuth nitrate, bismuth subnitrate, or bismuth chloride. In some embodiments, the thiol-containing compound comprises one or more agents selected from 1,2-ethanedithiol, 2,3-dimercaptopropanol, pyrithione, dithioerythritol, 3,4 dimercaptotoluene, 2,3-butanedithiol, 1,3-propanedithiol, 2-hydroxypropanethiol, 1-mercapto-2-propanol, dithioerythritol, dithiothreitol, and α-lipoic acid. In some embodiments, at least 70% of the microparticles have a volume average diameter of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm, or any narrower range between the specific ranges listed above.

[0147] In some embodiments, the BT composition comprises one or more BT compounds selected from the following:

[0148] [Table A-1]

[0149] [Table A-2]

[0150] In some embodiments, the bismuth thiol compound is BisEDT, having the following structure:

[0151] [ka] In some embodiments of the method of treating a topical wound, the BT composition administered is at least 1 μg / cm, including all integers and ranges therebetween. 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~Approx. 10,000μg / cm 2 In some embodiments, the administered BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~about 200μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 250 μg / cm 2 ~Approx. 5,000μg / cm 2 For example, in some embodiments, the bismuth thiol compound in the BT composition is present on the surface at a concentration of about 1 μg / cm 2 ~Approx. 10,000μg / cm 2 or about 50 μg / cm 2 ~about 200μg / cm 2 or 250 μg / cm 2 ~Approx. 5,000μg / cm 2 In some embodiments, the BT composition is present on the surface at a concentration of about 1 μg / cm 2 , about 50μg / cm 2 , about 100μg / cm 2 , about 150μg / cm 2 , about 200μg / cm 2 , about 250μg / cm 2 , about 500μg / cm 2 , about 750μg / cm 2 , about 1000μg / cm 2 , about 1500μg / cm 2 , about 2000μg / cm 2 , about 2500μg / cm 2 , about 3000μg / cm 2 , about 3500μg / cm 2 , about 4000μg / cm 2 , about 4500μg / cm 2 , about 5000μg / cm2 , about 5500μg / cm 2 , about 6000μg / cm 2 , about 6500μg / cm 2 , about 7000μg / cm 2 , about 7500μg / cm 2 , about 8000μg / cm 2 , about 8500μg / cm 2 , about 9000μg / cm 2 , about 9500μg / cm 2 , about 10,000μg / cm 2 It is present on surfaces in concentrations up to

[0152] In some embodiments, the present disclosure provides a method of treating or preventing a bacterial infection in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition described herein (e.g., a composition comprising BisEDT). In some embodiments, the bacterial infection is selected from the group consisting of Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp, Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, Coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium acnes, S. dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anaerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis. In some embodiments, the pharmaceutical composition is administered topically.In some embodiments, the subject is a mammal, e.g., a human. In some embodiments, the bacterial infection is a diabetic foot infection. In some embodiments, the diabetic foot infection comprises a skin ulcer. In some embodiments, the bacterial infection is associated with an area of ​​injured skin selected from wounds associated with cellulitis, erysipelas lesions, burns, chronic ulcers, bedsores, and pressure sores. In some embodiments, the treatment comprises topically administering the pharmaceutical composition to the skin ulcer associated with the diabetic foot infection. In some embodiments, the administration is followed by mechanical debridement of the ulcer. In some embodiments, the administration comprises the use of at least one of a bandage, an infusion device, and a negative pressure wound therapy device.

[0153] In some embodiments of the method of treating a local wound, the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months. In some embodiments, the BT composition is administered once a day or three times a week. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time greater than about 12 weeks. For example, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 1 year. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 4 weeks to about 10 weeks. In some embodiments, the pharmaceutical composition is administered every 4 hours or every 6 hours for the first 24 hours. In some embodiments, following the first 24 hours, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least three additional days, in some embodiments, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least four additional days.

[0154] In some embodiments of the method of treating a topical wound, the wound area is less than about 0.1 cm, including all integers and ranges therebetween. 2 ~about 250cm 2is. For example, the wound area is about 0.1 cm 2 , about 0.5 cm 2 , about 1 cm 2 , about 5 cm 2 , about 10 cm 2 , about 15 cm 2 , about 20 cm 2 , about 25 cm 2 , about 30 cm 2 , about 35 cm 2 , about 40 cm 2 , about 45 cm 2 , about 50 cm 2 , about 55 cm 2 , about 60 cm 2 , about 65 cm 2 , about 70 cm 2 , about 75 cm 2 , about 80 cm 2 , about 85 cm 2 , about 90 cm 2 , about 95 cm 2 , about 100 cm 2 , about 105 cm 2 , about 110 cm 2 , about 115 cm 2 , about 120 cm 2 , about 125 cm 2 , about 130 cm 2 , about 135 cm 2 , about 140 cm 2 , about 145 cm 2 , about 150 cm 2 , about 155 cm 2 , about 160 cm 2 , about 165 cm 2 , about 170 cm 2 , about 175 cm 2 , about 180 cm 2 , about 185 cm 2 , about 190 cm 2 , about 195 cm 2 , about 200 cm 2 , about 205 cm 2 , about 210 cm 2 , about 215 cm 2 , about 220 cm 2 , about 225 cm 2 , about 230 cm 2 of 235 cm 2 , about 240 cm2 , approx. 245cm 2 , or about 250 cm 2 may be also possible.

[0155] In some embodiments, the present disclosure provides a method of treating a bacterial infection comprising administering to a subject in need thereof a therapeutically effective amount of a composition comprising a bismuth thiol compound, where the composition is applied to the infection (e.g., applied to the surface of the infection). In some embodiments, the infection is a diabetic foot infection.In some embodiments, the bacterial infection is caused by any of the following bacterial pathogens: Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp, Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, Coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium acnes, S.dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis.

[0156] In some embodiments of the methods of treating a microbial (e.g., bacterial and / or fungal) infection, the infection is associated with a wound (e.g., an ulcer) and the subject experiences one or more of the following outcomes after completion of dosing: the wound is healed or substantially healed within 12 weeks (e.g., within 4 weeks) of the initial administration of the composition; and / or Preventing amputations and / or infection-related surgeries; and / or The wound closes partially or completely; and / or the wound is reduced in size from about 1% reduction compared to the original wound size to complete elimination of the wound; and / or The wound is more than 30 days old and is healed or substantially healed. In some embodiments, the subject experiences two or more of the recited results. In some embodiments, the subject experiences three or more of the recited results. In some embodiments, the subject experiences four or more of the recited results. In some embodiments, the subject experiences all of the recited results.

[0157] In some embodiments of the method of treating a microbial (e.g., bacterial and / or fungal) infection, the BT composition comprises one or more BT compounds selected from BisBAL, BisEDT, bis-dimercaprol, BisDTT, bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, BisPyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, bismuth-1-mercapto-2-propanol, and bis-EDT / 2-hydroxy-1-propanethiol. In some embodiments, the BT compound is BisEDT. In some embodiments, the composition is a suspension of microparticles comprising the BT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, at least 70% of the microparticles have a volume average diameter of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm, or any narrower range between the specific ranges listed above.

[0158] In some embodiments of the method of treating a microbial (e.g., bacterial and / or fungal) infection, the BT composition comprises BisEDT, and the BisEDT applied is about 20 μg / cm 2 present on surfaces in concentrations exceeding

[0159] In some embodiments of the method of treating a microbial (e.g., bacterial and / or fungal) infection, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4.

[0160] In some embodiments of the method of treating a microbial (e.g., bacterial and / or fungal) infection, the method comprises at least one of (i) reducing and / or dispersing a microbial (e.g., bacterial and / or fungal) biofilm, (ii) impairing the growth or formation of a microbial (e.g., bacterial and / or fungal) biofilm, and (iii) preventing the reformation or spread of a microbial (e.g., bacterial and / or fungal) biofilm. In some embodiments, the BT composition treats, manages, or reduces the severity of a diabetic foot infection by one or both of (i) preventing infection by bacterial or fungal pathogens and (ii) reducing bacterial or fungal pathogens. In some embodiments, the BT composition treats, manages, or reduces the severity of an infection by one or more of: (i) preventing the production or secretion of exotoxins from bacterial or fungal pathogens; (ii) inhibiting cell viability or cell proliferation of planktonic cells of bacterial or fungal pathogens; (iii) inhibiting the formation of biofilms or microbial pathogens by bacterial or fungal pathogens; (iv) inhibiting the invasiveness of biofilms into underlying tissues (e.g., subcutaneous tissue); (v) inhibiting the virulence of biofilms or microbial pathogens on underlying tissues (e.g., subcutaneous tissue); (vi) inhibiting biofilm viability or biofilm growth of biofilm-forming cells of bacterial or fungal pathogens; and / or (vii) preventing the reformation of biofilms after debridement.

[0161] In some embodiments of the methods of treating a microbial (e.g., bacterial and / or fungal) infection, the applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~Approx. 10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 250 μg / cm 2 ~Approx. 5,000μg / cm2 For example, in some embodiments, the bismuth thiol compound in the BT composition is present on the surface at a concentration of about 1 μg / cm 2 ~Approx. 10,000μg / cm 2 or about 50 μg / cm 2 ~about 200μg / cm 2 or 250 μg / cm 2 ~Approx. 5,000μg / cm 2 In some embodiments, the BT composition is present on the surface at a concentration of about 1 μg / cm 2 , about 50μg / cm 2 , about 100μg / cm 2 , about 150μg / cm 2 , about 200μg / cm 2 , about 250μg / cm 2 , about 500μg / cm 2 , about 750μg / cm 2 , about 1000μg / cm 2 , about 1500μg / cm 2 , about 2000μg / cm 2 , about 2500μg / cm 2 , about 3000μg / cm 2 , about 3500μg / cm 2 , about 4000μg / cm 2 , about 4500μg / cm 2 , about 5000μg / cm 2 , about 5500μg / cm 2 , about 6000μg / cm 2 , about 6500μg / cm 2 , about 7000μg / cm 2 , about 7500μg / cm 2 , about 8000μg / cm 2 , about 8500μg / cm 2 , about 9000μg / cm 2 , about 9500μg / cm 2 , approximately 10,000 μg / cm 2 It is present on surfaces in concentrations up to

[0162] In another embodiment, the dose volume may range from about 0.01 mL to about 10 mL, or any range therebetween. In another embodiment, the dose volume may range from about 0.1 mL to about 1 mL, or any range therebetween. In another embodiment, the minimum dose volume is from about 0.1 mL to about 0.5 mL, or any range therebetween.

[0163] In some embodiments of the method of treating a microbial (e.g., bacterial and / or fungal) infection, the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every other month. In some embodiments, the BT composition is administered once a day or three times a week. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about one week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time greater than about 12 weeks. For example, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 1 year. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time of about 10 weeks. In some embodiments, the pharmaceutical composition is administered within the first 24 hours, every 4 hours, or every 6 hours. In some embodiments, following the first 24 hours, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least three additional days, in some embodiments, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least four additional days.

[0164] In some embodiments of the methods of treating a microbial (e.g., bacterial and / or fungal) infection, the wound area is about 0.1 cm to about 250 cm. 2 For example, the wound area is about 0.1 cm 2 , about 0.5cm 2 , about 1cm 2 , about 5cm 2 , about 10cm 2 , about 15cm 2 , about 20cm 2 , about 25cm 2 , about 30cm2 , about 35cm 2 , about 40cm 2 , about 45cm 2 , about 50cm 2 , approx. 55cm 2 , about 60cm 2 , approx. 65cm 2 , about 70cm 2 , about 75cm 2 , about 80cm 2 , approx. 85cm 2 , about 90cm 2 , about 95cm 2 , about 100cm 2 , about 105cm 2 , about 110cm 2 , about 115cm 2 , about 120cm 2 , about 125cm 2 , about 130cm 2 , about 135cm 2 , about 140cm 2 , about 145cm 2 , about 150cm 2 , about 155cm 2 , about 160cm 2 , about 165cm 2 , about 170cm 2 , about 175cm 2 , about 180cm 2 , about 185cm 2 , about 190cm 2 , about 195cm 2 , about 200cm 2 , about 205cm 2 , about 210cm 2 , about 215cm 2 , about 220cm 2 , about 225cm 2 , about 230cm 2 , 235cm 2 , about 240cm 2 , approx. 245cm 2 Or about 250cm 2 may be also possible.

[0165] In some embodiments, the disclosure provides a method of healing a wound in a subject having a diabetic foot infection comprising administering to the subject a therapeutically effective amount of a composition comprising BisEDT, wherein the composition is a suspension of microparticles comprising said BisEDT, at least 70% of the microparticles having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm, the composition is applied to the infection (e.g., applied to the surface of the infection), and the wound is healed or substantially healed within 12 weeks of initial administration of the composition. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0166] In some embodiments of the method of healing a wound in a subject with a diabetic foot infection, the applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~Approx. 10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 For example, in some embodiments, the bismuth thiol compound in the BTS composition is present on the surface at a dosage of about 1 μg / cm. 2 ~Approx. 10,000μg / cm 2 or about 50 μg / cm 2 ~about 200μg / cm 2 or 250 μg / cm 2 ~Approx. 5,000μg / cm 2 In some embodiments, the BT composition is present on the surface at a concentration of about 1 μg / cm 2 , about 50μg / cm2 , about 100μg / cm 2 , about 150μg / cm 2 , about 200μg / cm 2 , about 250μg / cm 2 , about 500μg / cm 2 , about 750μg / cm 2 , about 1000μg / cm 2 , about 1500μg / cm 2 , about 2000μg / cm 2 , about 2500μg / cm 2 , about 3000μg / cm 2 , about 3500μg / cm 2 , about 4000μg / cm 2 , about 4500μg / cm 2 , about 5000μg / cm 2 , about 5500μg / cm 2 , about 6000μg / cm 2 , about 6500μg / cm 2 , about 7000μg / cm 2 , about 7500μg / cm 2 , about 8000μg / cm 2 , about 8500μg / cm 2 , about 9000μg / cm 2 , about 9500μg / cm 2 , approximately 10,000 μg / cm 2 It is present on surfaces in concentrations up to

[0167] In some embodiments of the method of healing a wound in a subject with a diabetic foot infection, the BT composition is administered 3 times a day, 2 times a day, 1 time a day, every other day, 1 time every 3 days, 3 times a week, 1 time a week, 1 time every other week, 1 time every month, or 1 time every other month. In some embodiments, the wound heals 4 weeks, 8 weeks, or 12 weeks after the first administration of the BT composition. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time longer than about 12 weeks. For example, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 1 year. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time of about 4 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of about 4 weeks to about 10 weeks. In some embodiments, the pharmaceutical composition is administered for the first 24 hours, every 4 hours, or every 6 hours. In some embodiments, following the first 24 hours, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least three additional days. In some embodiments, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least four additional days.

[0168] In some embodiments of the method of healing a wound in a subject with a diabetic foot infection, the wound area is about 0.1 cm 2 ~about 250cm 2 For example, the wound area is about 0.1 cm 2 , about 0.5cm 2 , about 1cm 2 , about 5cm 2 , about 10cm 2 , about 15cm 2 , about 20cm 2 , about 25cm 2 , about 30cm 2 , about 35cm 2 , about 40cm 2 , about 45cm 2 , about 50cm 2 , approx. 55cm 2, about 60cm 2 , approx. 65cm 2 , about 70cm 2 , about 75cm 2 , about 80cm 2 , approx. 85cm 2 , about 90cm 2 , about 95cm 2 , about 100cm 2 , about 105cm 2 , about 110cm 2 , about 115cm 2 , about 120cm 2 , about 125cm 2 , about 130cm 2 , about 135cm 2 , about 140cm 2 , about 145cm 2 , about 150cm 2 , about 155cm 2 , about 160cm 2 , about 165cm 2 , about 170cm 2 , about 175cm 2 , about 180cm 2 , about 185cm 2 , about 190cm 2 , about 195cm 2 , about 200cm 2 , about 205cm 2 , about 210cm 2 , about 215cm 2 , about 220cm 2 , about 225cm 2 , about 230cm 2 235cm 2 and about 240cm 2 , approx. 245cm 2 Or about 250cm 2 may be also possible.

[0169] In some embodiments, the disclosure provides a method of reducing the risk of amputation and / or infection-related surgery in a subject having a diabetic foot infection comprising administering to the subject a composition comprising a therapeutically effective amount of BisEDT, wherein the composition is applied to the infection (e.g., applied to the surface of the infection), and the risk of amputation and / or infection-related surgery is reduced by about 1% to about 100% compared to a similarly situated subject not treated with a composition comprising a therapeutically effective amount of a bismuth thiol compound. In some embodiments, the composition is a suspension of microparticles comprising said BisEDT, at least 70% of the microparticles having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4.

[0170] In some embodiments, the method of reducing the risk of amputation and / or infection-related surgery in a subject with a diabetic foot infection comprises administering about 1 μg / cm of the BT composition. 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 For example, in some embodiments, the bismuth thiol compound in the BTS composition is present on the surface at a dosage of about 1 μg / cm. 2 ~Approx. 10,000μg / cm 2 or about 50 μg / cm 2 ~about 200μg / cm 2 or 250 μg / cm 2 ~Approx. 5,000μg / cm2 In some embodiments, the BT composition is present on the surface at a concentration of about 1 μg / cm 2 , about 50μg / cm 2 , about 100μg / cm 2 , about 150μg / cm 2 , about 200μg / cm 2 , about 250μg / cm 2 , about 500μg / cm 2 , about 750μg / cm 2 , about 1000μg / cm 2 , about 1500μg / cm 2 , about 2000μg / cm 2 , about 2500μg / cm 2 , about 3000μg / cm 2 , about 3500μg / cm 2 , about 4000μg / cm 2 , about 4500μg / cm 2 , about 5000μg / cm 2 , about 5500μg / cm 2 , about 6000μg / cm 2 , about 6500μg / cm 2 , about 7000μg / cm 2 , about 7500μg / cm 2 , about 8000μg / cm 2 , about 8500μg / cm 2 , about 9000μg / cm 2 , about 9500μg / cm 2 , about 10,000μg / cm 2 It is present on surfaces in concentrations up to

[0171] In some embodiments, the method of reducing the risk of amputation and / or infection-related surgery in a subject with a diabetic foot infection comprises administering the BT composition three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months. In some embodiments, the BT composition is administered once a day or three times a week. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about one week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time greater than about 12 weeks. For example, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 1 year. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 4 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a period of about 4 weeks to about 10 weeks. In some embodiments, the pharmaceutical composition is administered for the first 24 hours, every 4 hours, or every 6 hours. In some embodiments, following the first 24 hours, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least three additional days. In some embodiments, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least four additional days.

[0172] In some embodiments, the method of reducing the risk of amputation and / or infection-related surgery in a subject with a diabetic foot infection comprises administering to a subject a wound having an area of ​​about 0.1 cm 2 ~about 250cm 2 For example, the wound area is about 0.1 cm 2 , about 0.5cm 2 , about 1cm 2 , about 5cm 2 , about 10cm 2 , about 15cm 2 , about 20cm 2 , about 25cm 2 , about 30cm 2 , about 35cm 2 , about 40cm 2 , about 45cm 2 , about 50cm 2, approx. 55cm 2 , about 60cm 2 , approx. 65cm 2 , about 70cm 2 , about 75cm 2 , about 80cm 2 , approx. 85cm 2 , about 90cm 2 , about 95cm 2 , about 100cm 2 , about 105cm 2 , about 110cm 2 , about 115cm 2 , about 120cm 2 , about 125cm 2 , about 130cm 2 , about 135cm 2 , about 140cm 2 , about 145cm 2 , about 150cm 2 , about 155cm 2 , about 160cm 2 , about 165cm 2 , about 170cm 2 , about 175cm 2 , about 180cm 2 , about 185cm 2 , about 190cm 2 , about 195cm 2 , about 200cm 2 , about 205cm 2 , about 210cm 2 , about 215cm 2 , about 220cm 2 , about 225cm 2 , about 230cm 2 235cm 2 and about 240cm 2 , approx. 245cm 2 Or about 250cm 2 may be also possible.

[0173] In some embodiments, the disclosure provides a method of closing a wound in a subject having a diabetic foot infection, comprising administering to the subject a therapeutically effective amount of a composition comprising BisEDT. In some embodiments, the composition is a suspension of microparticles comprising said BisEDT, wherein at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4.

[0174] In some embodiments, in the method of closing a wound in a subject with a diabetic foot infection, the applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~Approx. 10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 For example, in some embodiments, the bismuth thiol compound in the BTS composition is present on the surface at a dosage of about 1 μg / cm. 2 ~Approx. 10,000μg / cm 2 or about 50 μg / cm 2 ~about 200μg / cm 2 or 250 μg / cm 2 ~Approx. 5,000μg / cm 2 In some embodiments, the BT composition is present on the surface at a concentration of about 1 μg / cm 2 , about 50μg / cm 2 , about 100μg / cm 2 , about 150μg / cm 2, about 200μg / cm 2 , about 250μg / cm 2 , about 500μg / cm 2 , about 750μg / cm 2 , about 1000μg / cm 2 , about 1500μg / cm 2 , about 2000μg / cm 2 , about 2500μg / cm 2 , about 3000μg / cm 2 , about 3500μg / cm 2 , about 4000μg / cm 2 , about 4500μg / cm 2 , about 5000μg / cm 2 , about 5500μg / cm 2 , about 6000μg / cm 2 , about 6500μg / cm 2 , about 7000μg / cm 2 , about 7500μg / cm 2 , about 8000μg / cm 2 , about 8500μg / cm 2 , about 9000μg / cm 2 , about 9500μg / cm 2 , approximately 10,000 μg / cm 2 It is present on surfaces in concentrations up to

[0175] In some embodiments, the method of closing a wound in a subject with a diabetic foot infection, the composition is applied to the infection (e.g., applied to the surface of the infection), and the wound closes within 12 weeks of the initial administration of the composition. In some embodiments, the BT composition is administered once a day or three times a week. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time greater than about 12 weeks. For example, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 1 year. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 4 weeks to about 10 weeks. In some embodiments, the pharmaceutical composition is administered every 4 hours or every 6 hours for the first 24 hours. In some embodiments, following the first 24 hours, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least three additional days, in some embodiments, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least four additional days.

[0176] In some embodiments, the method of closing a wound in a subject with a diabetic foot infection comprises: 2 ~about 250cm 2 For example, the wound area is about 0.1 cm 2 , about 0.5cm 2 , about 1cm 2 , about 5cm 2 , about 10cm 2 , about 15cm 2 , about 20cm 2 , about 25cm 2 , about 30cm 2 , about 35cm 2 , about 40cm 2 , about 45cm 2 , about 50cm 2 , approx. 55cm 2 , about 60cm 2, approx. 65cm 2 , about 70cm 2 , about 75cm 2 , about 80cm 2 , approx. 85cm 2 , about 90cm 2 , about 95cm 2 , about 100cm 2 , about 105cm 2 , about 110cm 2 , about 115cm 2 , about 120cm 2 , about 125cm 2 , about 130cm 2 , about 135cm 2 , about 140cm 2 , about 145cm 2 , about 150cm 2 , about 155cm 2 , about 160cm 2 , about 165cm 2 , about 170cm 2 , about 175cm 2 , about 180cm 2 , about 185cm 2 , about 190cm 2 , about 195cm 2 , about 200cm 2 , about 205cm 2 , about 210cm 2 , about 215cm 2 , about 220cm 2 , about 225cm 2 , about 230cm 2 235cm 2 and about 240cm 2 , approx. 245cm 2 Or about 250cm 2 may be also possible.

[0177] In some embodiments, the disclosure provides a method for wound size reduction in a subject with a diabetic foot infection, comprising administering to the subject a therapeutically effective amount of a composition comprising BisEDT, wherein the composition is applied to the infection (e.g., applied to the surface of the infection), and within 12 weeks of the initial administration of the composition, the wound is reduced in size from about 1% reduction compared to the original wound size to complete clearance of the wound. In some embodiments, the wound is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%. In some embodiments, the wound is reduced by about 50%.

[0178] In some embodiments, the method for wound size reduction in a subject with a diabetic foot infection comprises a composition comprising a suspension of microparticles comprising BisEDT, wherein at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0179] In some embodiments, in the method for reducing wound size in a subject with a diabetic foot infection, the applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~Approx. 10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 For example, in some embodiments, the bismuth thiol compound in the BTS composition is present on the surface at a dosage of about 1 μg / cm.2 ~Approx. 10,000μg / cm 2 or about 50 μg / cm 2 ~about 200μg / cm 2 or 250 μg / cm 2 ~Approx. 5,000μg / cm 2 In some embodiments, the BT composition is present on the surface at a concentration of about 1 μg / cm 2 , about 50μg / cm 2 , about 100μg / cm 2 , about 150μg / cm 2 , about 200μg / cm 2 , about 250μg / cm 2 , about 500μg / cm 2 , about 750μg / cm 2 , about 1000μg / cm 2 , about 1500μg / cm 2 , about 2000μg / cm 2 , about 2500μg / cm 2 , about 3000μg / cm 2 , about 3500μg / cm 2 , about 4000μg / cm 2 , about 4500μg / cm 2 , about 5000μg / cm 2 , about 5500μg / cm 2 , about 6000μg / cm 2 , about 6500μg / cm 2 , about 7000μg / cm 2 , about 7500μg / cm 2 , about 8000μg / cm 2 , about 8500μg / cm 2 , about 9000μg / cm 2 , about 9500μg / cm 2 , about 10,000μg / cm 2 It is present on surfaces in concentrations up to

[0180] In some embodiments, the method for wound size reduction in a subject with a diabetic foot infection includes administering the BT composition three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every other month. In some embodiments, the BT composition is administered once a day or three times a week. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about one week to about 12 weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about four weeks to about ten weeks. In some embodiments, the subject is administered multiple doses of the BT composition daily or weekly for a length of time greater than about 12 weeks. For example, about four months, about five months, about six months, about seven months, about eight months, about nine months, about ten months, about eleven months, or about one year. In some embodiments, the pharmaceutical composition is administered for the first 24 hours, every 4 hours, or every 6 hours. In some embodiments, following the first 24 hours, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least an additional 3 days. In some embodiments, the pharmaceutical composition is administered every 12 hours or every 24 hours for at least an additional 4 days.

[0181] In some embodiments, the method for reducing wound size in a subject with a diabetic foot infection comprises administering to a subject a wound having an area of ​​about 0.1 cm 2 ~about 250cm 2 For example, the wound area is about 0.1 cm 2 , about 0.5cm 2 , about 1cm 2 , about 5cm 2 , about 10cm 2 , about 15cm 2 , about 20cm 2 , about 25cm 2 , about 30cm 2 , about 35cm 2 , about 40cm 2 , about 45cm 2 , about 50cm 2 , approx. 55cm 2, about 60cm 2 , approx. 65cm 2 , about 70cm 2 , about 75cm 2 , about 80cm 2 , approx. 85cm 2 , about 90cm 2 , about 95cm 2 , about 100cm 2 , about 105cm 2 , about 110cm 2 , about 115cm 2 , about 120cm 2 , about 125cm 2 , about 130cm 2 , about 135cm 2 , about 140cm 2 , about 145cm 2 , about 150cm 2 , about 155cm 2 , about 160cm 2 , about 165cm 2 , about 170cm 2 , about 175cm 2 , about 180cm 2 , about 185cm 2 , about 190cm 2 , about 195cm 2 , about 200cm 2 , about 205cm 2 , about 210cm 2 , about 215cm 2 , about 220cm 2 , about 225cm 2 , about 230cm 2 235cm 2 and about 240cm 2 , approx. 245cm 2 Or about 250cm 2 In some embodiments, the wound surface area of ​​the wound is reduced by 50% 12 weeks after the first administration of the BT composition, the BT composition being BisEDT. In some embodiments, the wound surface area of ​​the wound is reduced by 50% 4 weeks after the first administration of the BisEDT composition. In some embodiments, the wound surface area is measured using digital photography or manual measurement.

[0182] In some embodiments, the present disclosure provides a method for preventing amputation and / or infection-related surgery in a subject with a diabetic foot infection, comprising administering to the subject a therapeutically effective amount of a BT composition. In some embodiments, the BT composition is a suspension of microparticles comprising the BisEDT, wherein at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4.

[0183] In some embodiments, in the method of preventing amputation and / or infection-related surgery in a subject with a diabetic foot infection, the applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 For example, in some embodiments, the bismuth thiol compound in the BTS composition is present on the surface at a dosage of about 1 μg / cm. 2 ~Approx. 10,000μg / cm 2 or about 50 μg / cm 2 ~about 200μg / cm 2 or 250 μg / cm 2 ~Approx. 5,000μg / cm 2 In some embodiments, the BT composition is present on the surface at a concentration of about 1 μg / cm. 2 , about 50μg / cm 2 , about 100μg / cm 2 , about 150μg / cm2 , about 200μg / cm 2 , about 250μg / cm 2 , about 500μg / cm 2 , about 750μg / cm 2 , about 1000μg / cm 2 , about 1500μg / cm 2 , about 2000μg / cm 2 , about 2500μg / cm 2 , about 3000μg / cm 2 , about 3500μg / cm 2 , about 4000μg / cm 2 , about 4500μg / cm 2 , about 5000μg / cm 2 , about 5500μg / cm 2 , about 6000μg / cm 2 , about 6500μg / cm 2 , about 7000μg / cm 2 , about 7500μg / cm 2 , about 8000μg / cm 2 , about 8500μg / cm 2 , about 9000μg / cm 2 , about 9500μg / cm 2 , about 10,000μg / cm 2 It is present on surfaces in concentrations up to

[0184] In some embodiments, the disclosure provides a method of treating a wound in a subject, wherein the wound is 30 days or older, comprising administering a therapeutically effective amount of a BT composition to the subject. In some embodiments, the subject has a diabetic foot infection. In some embodiments, the BT composition is a suspension of microparticles comprising BisEDT, wherein at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4.

[0185] In some embodiments, the disclosure provides a method of treating a wound in a subject, wherein the wound is within 30 days of administration of a therapeutically effective amount of a BT composition to the subject. In some embodiments, the subject has a diabetic foot infection. In some embodiments, the BT composition is a suspension of microparticles comprising BisEDT having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4. For example, the wound may be about the 1st day, about the 2nd day, about the 3rd day, about the 4th day, about the 5th day, about the 6th day, about the 7th day, about the 8th day, about the 9th day, about the 10th day, about the 11th day, about the 12th day, about the 13th day, about the 14th day, about the 15th day, about the 16th day, about the 17th day, about the 18th day, about the 19th day, about the 20th day, about the 21st day, about the 22nd day, about the 23rd day, about the 24th day, about the 25th day, about the 26th day, about the 27th day, about the 28th day, about the 29th day, or about the 30th day.

[0186] In some embodiments, in the method of treating a wound in a subject, the wound is 30 days or older, the BT composition applied is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~Approx. 10,000μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 For example, in some embodiments, the bismuth thiol compound in the BTS composition is present on the surface at a dosage of about 1 μg / cm. 2 ~Approx. 10,000μg / cm 2 or about 50 μg / cm 2~about 200μg / cm 2 or 250 μg / cm 2 ~Approx. 5,000μg / cm 2 In some embodiments, the BT composition is present on the surface at a concentration of about 1 μg / cm 2 , about 50μg / cm 2 , about 100μg / cm 2 , about 150μg / cm 2 , about 200μg / cm 2 , about 250μg / cm 2 , about 500μg / cm 2 , about 750μg / cm 2 , about 1000μg / cm 2 , about 1500μg / cm 2 , about 2000μg / cm 2 , about 2500μg / cm 2 , about 3000μg / cm 2 , about 3500μg / cm 2 , about 4000μg / cm 2 , about 4500μg / cm 2 , about 5000μg / cm 2 , about 5500μg / cm 2 , about 6000μg / cm 2 , about 6500μg / cm 2 , about 7000μg / cm 2 , about 7500μg / cm 2 , about 8000μg / cm 2 , about 8500μg / cm 2 , about 9000μg / cm 2 , about 9500μg / cm 2 , about 10,000μg / cm 2 It is present on surfaces in concentrations up to

[0187] In some embodiments, in the method of treating a wound in a subject, the wound is less than 30 days old, the applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 (For example, about 1 μg / cm 2 ~Approx. 10,000μg / cm 2In some embodiments, the applied BT composition is present on the surface at a concentration of about 50 μg / cm 2 ~Approx. 100μg / cm 2 In some embodiments, the applied BT composition is present on the surface at a concentration of about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 For example, in some embodiments, the bismuth thiol compound in the BTS composition is present on the surface at a dosage of about 1 μg / cm. 2 ~Approx. 10,000μg / cm 2 or about 50 μg / cm 2 ~about 200μg / cm 2 or 250 μg / cm 2 ~Approx. 5,000μg / cm 2 In some embodiments, the BT composition is present on the surface at a concentration of about 1 μg / cm. 2 , about 50μg / cm 2 , about 100μg / cm 2 , about 150μg / cm 2 , about 200μg / cm 2 , about 250μg / cm 2 , about 500μg / cm 2 , about 750μg / cm 2 , about 1000μg / cm 2 , about 1500μg / cm 2 , about 2000μg / cm 2 , about 2500μg / cm 2 , about 3000μg / cm 2 , about 3500μg / cm 2 , about 4000μg / cm 2 , about 4500μg / cm 2 , about 5000μg / cm 2 , about 5500μg / cm 2 , about 6000μg / cm 2 , about 6500μg / cm 2 , about 7000μg / cm 2 , about 7500μg / cm 2 , about 8000μg / cm 2 , about 8500μg / cm 2, about 9000μg / cm 2 , about 9500μg / cm 2 , about 10,000μg / cm 2 It is present on surfaces in concentrations up to

[0188] In some embodiments, the method of treating a wound in a subject, the wound is more than 30 days old, the wound is more than 2 months old, more than 3 months old, more than 4 months old, more than 5 months old, more than 6 months old, more than 7 months old, more than 8 months old, more than 9 months old, more than 10 months old, more than 11 months old, or more than 1 year old. In some embodiments, the wound is more than 2 months old. In some embodiments, the wound is more than 3 months old.

[0189] In any of the embodiments of the methods described herein, the BT composition ultimately applied or administered to the subject may be from about 1 μg / mL to about 1,000,000 μg / mL (e.g., about 1 μg / cm 2 ~Approx. 10,000μg / cm 2) in the presence of 50 μg / mL to about 100 μg / mL. In some embodiments, the BT composition applied has a concentration of about 250 μg / mL to about 5,000 μg / mL. For example, in some embodiments, the bismuth thiol compound in the BT composition is BisEDT having a concentration of about 1 μg / mL to about 10,000 μg / mL, or about 50 μg / mL to about 200 μg / mL, or about 250 μg / mL to about 5,000 μg / mL. In some embodiments, the BT composition is about 1 μg / mL, about 50 μg / mL, about 100 μg / mL, about 150 μg / mL, about 200 μg / mL, about 250 μg / mL, about 500 μg / mL, about 750 μg / mL, about 1000 μg / mL, about 1500 μg / mL, about 2000 μg / mL, about 2500 μg / mL, about 3000 μg / mL, about 3500 μg / mL, about 4000 μg / mL, about 4500 μg / mL, about 5000 μg / mL, about 6000 μg / mL, about 7500 μg / mL, about 1000 μg / mL, about 1500 μg / mL, about 2000 μg / mL, about 2500 μg / mL, about 3000 μg / mL, about 3500 μg / mL, about 4000 μg / mL, about 5000 μg / mL, about 6500 μg / mL, about 7000 μg / mL, about 8000 μg / mL, about 8500 μg / mL, about 9000 μg / mL, about 9000 μg / mL, about 10000 μg / mL, about 11000 μg / mL, about 12000 μg / mL, about 13000 μg / mL, about 14000 μg / mL, about 15000 μg / mL, about 16000 μg / mL, about 17000 μg / mL, about 18000 μg / mL, about 19000 μg / mL, about 20000 μg / mL, about 21000 μg / mL, about 2200 / mL, about 4000 μg / mL, about 4500 μg / mL, about 5000 μg / mL, about 5500 μg / mL, about 6000 μg / mL, about 6500 μg / mL, about 7000 μg / mL, about 7500 μg / mL, about 8000 μg / mL, about 8500 μg / mL, about 9000 μg / mL, about 9500 μg / mL, about 10,000 μg / mL. In some embodiments of the methods described herein, the BT composition that is ultimately applied or administered to the subject has a concentration of more than about 1,000,000 μg / mL.

[0190] In certain embodiments, the present invention may be a pharmaceutical composition comprising a bismuth thiol (BT) composition comprising BisEDT suspended therein, wherein the BT composition comprises a plurality of microparticles. In certain embodiments, the microparticles have a D90 of 4.5 μm, or 4.0 μm, or 3.5 μm, or 3.0 μm, or 2.5 μm, or 2.0 μm, or 1.9 μm, or 1.8 μm, or 1.7 μm, or 1.6 μm, or 1.5 μm or less, or any range therebetween. In a specific embodiment, the microparticles have a D90 of 1.9 μm or less. In another specific embodiment, the microparticles have a D90 of 1.6 μm or less. In another specific embodiment, the microparticle has a D50 of 2.5 μm, or 2.0 μm, or 1.5 μm, or 1.3 μm, or 1.2 μm, or 1.1 μm, or 1.0 μm, or 0.9 μm, or 0.87 μm, or 0.72 μm, or any range therebetween. In another specific embodiment, the microparticle has a D10 of 0.9 μm, or 0.8 μm, or 0.7 μm, or 0.6 μm, or 0.50 μm, or 0.40 μm, or 0.39 μm, or 0.38 μm, or 0.37 μm, or 0.36 μm, or 0.35 μm, or 0.34 μm, or 0.33 μm, or any range therebetween. In a specific embodiment, a pharmaceutical composition comprising a bismuth thiol (BT) composition comprises BisEDT suspended therein, wherein the BT composition comprises a plurality of microparticles, the microparticles having a D90 of about 1.6 μm or less. In a specific embodiment, the BT composition comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7. In another specific embodiment, the composition described above can be administered to a subject to treat a topical wound in a subject or in any of the specific methods described herein. In another specific embodiment, the composition described above can be administered to a subject to treat a diabetic foot infection or a diabetic foot ulcer.In specific embodiments, the method may include administering to a subject a composition as described above, wherein the subject experiences one or more of the following results after completion of dosing of said composition: a) the wound is healed or substantially healed within 12 weeks of the initial administration of the composition; and / or b) preventing amputations and / or infection-related surgery; and / or c) the wound is partially or completely closed; and / or d) the wound is reduced in size from approximately 1% reduction in size compared to the original wound size to complete disappearance of the wound; and / or e) The wound is at least 30 days old and is healed or substantially healed. Combination treatment In some embodiments, the present disclosure provides a method for treating and / or preventing diabetic foot infections, comprising administering the BT composition of the present disclosure in combination with standard and / or non-standard treatments for diabetic foot infections. Standard treatments for diabetic foot infections, including but not limited to the associated skin ulcers, include extracellular matrix replacement therapy (including the use of autologous cells), moist wound therapy, negative pressure wound therapy, arterial revascularization therapy, hyperbaric oxygen therapy, administration of antimicrobial agents, administration of antibiotics, topical oxygen therapy, photodynamic therapy, high energy acoustic shock wave therapy, ultrasound therapy, and administration of growth factors (Blume, et al. 2008, Diabetes Care, 31:631-636). In some embodiments, the treatment is negative pressure wound therapy. As used herein, the terms "in combination" or "in further combination" or "further in combination" refer to the use of additional prophylactic and / or therapeutic agents, as well as the BT composition of the present disclosure.

[0191] The use of the term "in combination" does not limit the order in which prophylactic and / or therapeutic agents are administered to a subject. A first prophylactic or therapeutic agent can be administered prior to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), simultaneously with, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of a second prophylactic or therapeutic agent (different from the first prophylactic or therapeutic agent) to a subject.

[0192] In some embodiments, the BT composition of the present disclosure is administered locally, e.g., at the site of a diabetic foot ulcer, while the additional agent is administered systemically. For example, in some embodiments, the BT composition of the present disclosure is administered locally, e.g., at the site of a diabetic foot ulcer, while the antibiotic is administered systemically.In further embodiments, including the treatment of diabetic foot ulcers, the antibiotic administered systemically is selected from the group consisting of Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp., Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, Coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium acnes, S. dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anaerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis. In some embodiments, the BT composition of the present disclosure is administered locally, for example, to the site of a diabetic foot ulcer, along with an additional agent that is also administered locally.For example, in some embodiments, the BT composition of the present disclosure is administered locally along with growth factors, for example, to the site of a diabetic foot ulcer.

[0193] In some embodiments, the method is used in combination with standard therapies for diabetic foot infections, such as standard therapies selected from the group consisting of extracellular matrix replacement therapy (including the use of autologous cells), moist wound therapy, negative pressure wound therapy, arterial revascularization therapy, hyperbaric oxygen therapy, administration of antimicrobial agents, administration of antibiotics, topical oxygen therapy, photodynamic therapy, high energy acoustic shock wave therapy, ultrasound therapy, and administration of growth factors (Blume et al. 2008, Diabetes Care, 31:631-636). In some embodiments, the therapy is negative pressure wound therapy. In some embodiments, the moist wound therapy includes the use of adhesive backing films, silicone coated foams, hydrogels, and / or hydrocolloids. In some embodiments, the extracellular matrix replacement therapy includes the use of bioengineered tissue. In some embodiments, the administration of antibiotics includes systemic administration. In some embodiments, the growth factor is at least one selected from the group consisting of platelet-derived growth factor, granulocyte colony stimulating factor, epidermal growth factor, fibroblast growth factor, nerve growth factor, and vascular endothelial growth factor. In some embodiments, administration of the growth factor comprises local administration. In some embodiments, the method is used in combination with a non-standard treatment for diabetic foot infection, for example, when the diabetic foot infection is refractory to standard therapy.

[0194] Extracellular matrix therapy is used to treat, manage, control, and / or prevent non-healing areas of wounded skin and is the standard of care for diabetic foot ulcers. Synthesis of the extracellular matrix (ECM) is a key feature of wound healing, especially when there is significant tissue loss. Extracellular matrix therapy is designed to reduce protease levels in wound fluid by providing a competitive substrate for the proteases (collagen), thereby reducing proteolytic destruction of essential ECM components and promoting healing. For example, ECM therapy may include administration of agents that reduce the synthesis of matrix metalloproteinases (MMPs), such as mixtures of metal cations, as well as reducing the proteolytic destruction of fibronectin and / or platelet-derived growth factor (PDGF). ECM therapy may also include administration of amelogenin, an ECM protein with biological activity in skin regeneration and repair (Romanelli M. 2010 Wounds-Clinical Review 6(2):47-52).

[0195] ECM therapy may also include the use of bioengineered tissues to replace lost ECM, for example. Bioengineered tissues, also called "skin replacement products" or "skin substitutes," often contain a biological matrix with or without living cells (Brian DL et al. 2011. Expert Rev Dermatol. 6(3):255-262). Most bioengineered tissues can be divided into living tissue substitutes versus bioactive adjuncts. Bioengineered tissues may look like thin, circular pieces of actual skin and can be placed directly onto areas of injured skin. While the exact mechanism of healing is not fully understood, bioengineered tissues are thought to improve healing by filling the wound with extracellular matrix proteins and possibly also expressing additional growth factors and cytokines that promote healing. Specific examples of bioengineered tissues used to treat diabetic foot ulcers include Apligraf and Dermagraft, which are commercially available.

[0196] Arterial revascularization therapy (ART) is also used to treat, manage, control, and / or prevent non-healing areas of damaged skin and is the standard of care for diabetic foot ulcers. The preferred approach in treating diabetic foot infections involves percutaneous methods of ART, which include percutaneous balloon angioplasty with or without stent placement. Other revascularization approaches include aortoiliac reconstruction with aortofemoral bypass and femoral-popliteal-tibial bypass using the saphenous vein.

[0197] Hyperbaric oxygen therapy (HBOT) is also used to treat, manage, control, and / or prevent non-healing areas of broken skin and is the standard treatment for diabetic foot ulcers. HBOT refers to the intermittent treatment of the whole body with pressures higher than normal atmospheric pressure, often resulting in increased oxygen content compared to normal air. For example, hyperbaric oxygen chambers may be used to increase pressure to up to twice normal atmospheric pressure. Patients may also be exposed to concentrations of up to 100% oxygen. The increased pressure, combined with increased oxygen content, dissolves oxygen in the plasma, body cells, tissues, and fluids, which in turn aids in the wound healing process. It is believed that HBOT can stimulate the growth of new blood vessels to areas of reduced circulation and improve blood flow to areas with blocked arteries.

[0198] In some other embodiments, the present disclosure provides methods for treating and / or preventing diabetic foot infections comprising administering a BT composition of the present disclosure in combination with a non-standard treatment for diabetic foot infections, which is generally used when a diabetic foot ulcer is refractory to one or more standard therapies.

[0199] In certain embodiments, the compositions disclosed herein can be used alone or in combination with another type of therapeutic agent.In certain embodiments, the BT compound is administered locally during any of the surgeries described herein, and systemic antibiotics are administered at regular intervals before or after surgery.In some embodiments, systemic antibiotics are administered before surgery.

[0200] In certain embodiments, the compositions of the present disclosure are administered in combination with one or more additional therapeutic agent(s) to provide improved efficacy compared to the individual administration of the compound of the present disclosure or the one or more additional therapeutic agent(s). In certain such embodiments, the co-administration provides an additive or synergistic effect, where additive effect refers to the sum of the effects of the individual administration of the compound of the present disclosure and the one or more additional therapeutic agent(s), respectively. In some embodiments, the subject receives co-administration of a treatment for another disease, disorder, or condition. In some embodiments, the other treatment is an immunosuppressive treatment.

[0201] Standard antibiotics that may be used with the compositions of the present disclosure include amikacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin, streptomycin, tobramycin, apramycin, rifamycin, naphthomycin, mupirocin, geldanamycin, ansamitocin, carbacephems, imipenem, meropenem, ertapenem, faropenem, doripenem, panipenem, erythromycin ... Nem / betamipron, biapenem, PZ-601, cephalosporin, cephacetrile, cefadroxil, cephalexin, cephaloglycin, cephalonium, cephaloridine, cephalothin, cephapirin, cefatrizine, cefazaflur, cefazedone, cefazolin, cephradine, cefroxadine, ceftezole, cefaclor, cefonicid, cefprozil, cefuroxime, cefuzonam, cefmetazole, cefotetan, cefoxitin, cefcapene, cefdaloxime, cefdinir, cefditoren, cefetamet, cefixime, cefmenon oxime, cefteram, ceftibuten, ceftiofur, ceftiolene, ceftizoxime, ceftriaxone, cefoperazone, ceftazidime, latamoxef, cefclidine, cefepime, cefluprenam, cefoselis, cefozopran, cefpirome, cefquinome, flomoxef, ceftobiprole, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, aztreonam, penicillin and penicillin derivatives, actinomycin, bacitracin, colistin, polymyxin B, cinoxacin, flumequine, Nalidixic acid, oxolinic acid, piromidic acid, pipemidic acid, losoxacin, ciprofloxacin, enoxacin, fleroxacin, lomefloxacin, nadifloxacin, norfloxacin, ofloxacin, pefloxacin, rufloxacin, balofloxacin, gatifloxacin, grepafloxacin, levofloxacin, moxifloxacin, pazufloxacin, sparfloxacin, temafloxacin, tosufloxacin, clinafloxacin, garenoxacin, gemifloxacin, stifloxacin, trovafloxacin, prulifloxacin,Acetazolamide, Benzolamide, Bumetanide, Celecoxib, Chlorthalidone, Clopamide, Dichlorphenamide, Dorzolamide, Ethoxolamide, Furosemide, Hydrochlorothiazine, Idapamide, Mafendin, Mefruside, Metolazone, Probenecid, Sulfacetamide, Sulfadimethoxine, Sulfadoxine, Sulfanilamide, Sulfamethoxazole, Sulfasalazine, Sulthiame, Sumatriptan, Xipamide, Tetracycline , chlortetracycline, oxytetracycline, doxycycline, lymecycline, meclocycline, methacycline, minocycline, rolitetracycline, methacillin, nafcillin, oxacillin, cloxacillin, vancomycin, teicoplanin, cylindamycin, co-trimoxazole, flucloxacillin, dicloxacillin, ampicillin, amoxicillin, and any combination thereof. In some embodiments, the methods of the disclosure include the use of amikacin, tobramycin, gentamicin, piperacillin, mezlocillin, ticarcillin, imipenem, ciprofloxacin, ceftazidime, aztreonam, ticaricillin clavulanate, dicloxacillin, amoxicillin, ticaricillin clavulanate, trimethoprim-sulfamethoxazole, cephalexin, piperacillin-tazobactam, linezolid, daptomycin, vancomycin, metronidazole, clindamycin, colistin, tetracycline, levofloxacin, amoxicillin and clavulanic acid (Augmentin®), clonidine, cyclosporine ... and co-administration to the subject of an antibiotic selected from sacillin, dicloxalin, cefdinir, cefprozil, cefaclor, cefuroxime, erythromycin / sulfisoxazole, erythromycin, clarithromycin, azithromycin, doxycycline, minocycline, tigecycline, imipenem, melipenem, colistimethate / colistin®, methicillin, oxacillin, nafcillin, cabenicillin, azlocillin, piperacillin, dalbavancin, oritavancin, tedizolid, delafloxacin, and tazobactam (Zosyn®), cefepime, and meropenem.

[0202] In certain embodiments of the present disclosure, therapeutic agents that can be co-administered with compounds of the present disclosure, such as bismuth thiol compounds, include known antibiotics. In some embodiments, the antibiotic is selected from methicillin, vancomycin, nafcillin, gentamicin, ampicillin, chloramphenicol, doxycycline, tobramycin, and levofloxacin. In some embodiments, the antibiotic is selected from tobramycin, imipenem, tetracycline, and minocycline. In some embodiments, the antibiotic is administered systemically after revision surgery. In some embodiments, the antibiotic is administered prior to revision surgery. Co-administered therapeutic agents, such as antibiotics, can be administered at any suitable frequency and in any suitable dosage. Such dosages and frequencies can be determined by one of skill in the art.

[0203] In certain embodiments, the BT compounds of the present disclosure can be administered in combination with one or more other BT compounds of the present disclosure. Additionally, such combinations can be administered in combination with other therapeutic agents. Pharmaceutical Compositions In some embodiments, the BT composition comprises one or more BT compounds selected from BisBAL, BisEDT, bis-dimercaprol, BisDTT, bis-2-mercaptoethanol, Bis-DTE, bis-Pyr, bis-Ery, bis-Tol, bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, BisPyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, bismuth-1-mercapto-2-propanol, and bis-EDT / 2-hydroxy-1-propanethiol, and the composition is a suspension of microparticles comprising the BT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm. In some embodiments, the BT compound is BisEDT.

[0204] In some embodiments, at least 70% of the microparticles have a volume average diameter of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm, or any narrower range between the specific ranges listed above.

[0205] In some embodiments, the BT composition comprises BisEDT at a concentration of greater than about 100 μg / mL. In some embodiments, the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate at about pH 7.4.

[0206] In certain embodiments, the BT composition is a suspension of one or more BT compounds in Tween (e.g., Tween 80) and / or buffer (e.g., sodium phosphate buffer). For example, in some embodiments, the BT composition is a suspension of one or more BT compounds in about 0.1% Tween 80 to about 1.0% Tween 80, including all ranges therebetween. For example, the BT composition is a suspension of one or more BT compounds in about 0.1% Tween 80, about 0.2% Tween 80, about 0.3% Tween 80, about 0.4% Tween 80, about 0.5% Tween 80, about 0.6% Tween 80, about 0.7% Tween 80, about 0.8% Tween 80, about 0.9% Tween 80, or about 1% Tween 80. In some embodiments, the BT composition is a suspension of one or more BT compounds in about 0.5% Tween 80. In some embodiments, the BT composition is a suspension of one or more BT compounds in about 1.1% Tween 80, about 1.2% Tween 80, about 1.3% Tween 80, about 1.4% Tween 80, about 1.5% Tween 80, about 1.6% Tween 80, about 1.7% Tween 80, about 1.8% Tween 80, about 1.9% Tween 80, or about 2% Tween 80.

[0207] The disclosed BT compositions and methods can be utilized to treat a subject in need thereof. In certain embodiments, the subject is a mammal, such as a human or a non-human mammal. When administered to a subject, such as a human, the composition or compound may be administered, for example, as a pharmaceutical composition comprising a compound of the disclosure and a pharma- ceutically acceptable carrier. Some examples of substances that can serve as pharma- ceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, methylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) propylene glycol; and (11) propylene glycol. (11) polyols and sugar alcohols such as glycerin, sorbitol, mannitol, xylitol, erythritol, and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances including salts such as sodium chloride used in pharmaceutical formulations. In some embodiments, when such pharmaceutical compositions are for human administration, the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to achieve delayed release of the drug or to selectively target one or more cells, tissues, or organs. The pharmaceutical composition can be in unit dosage form such as a lyophilisate, powder, solution, syrup, infusion, etc. for reconstitution. The composition can also be in a solution suitable for topical administration.

[0208] In some embodiments, the pharmaceutical compositions of the present disclosure are formulated for topical administration, for example, to areas of damaged skin. Damaged skin can include, but is not limited to, skin lesions, blisters, chronic ulcers, cysts, blisters, blisters, open ulcers such as bedsores (pressure sores) and other pressure sores, cellulitis wounds, erysipelas lesions, wounds, burns, pimples, skin ulcers, such as those associated with diabetic foot infections, or other conditions in which the skin is damaged, broken, cracked, torn, and / or otherwise compromised. Topical formulations generally include a sterile buffer, such as sterile PBS, water, or saline buffer, or a sterile SM buffer. A variety of buffers may be used in the context of the present disclosure, and will be readily apparent to those skilled in the art. For example, in some embodiments, suitable buffers include sodium or potassium citrate, citric acid, phosphate buffers such as sodium phosphate, boric acid, sodium bicarbonate, and various mixed phosphate buffers including combinations of Na2HPO4, NaH2PO4, and KH2PO4. In some embodiments, sodium phosphate buffers are used. In some embodiments, sodium citrate buffers are used. Thus, the pH of the formulation may vary from about 5 to about 10. In some embodiments, the pH of the formulation is about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, or about 10. In some embodiments, the pH of the formulation is about 7.4.

[0209] In some embodiments, the BT composition is a suspension of one or more BT compounds in about 0.5% Tween 80 in sodium phosphate buffer at a pH of about 7.4. In some embodiments, the one or more BT compounds are present in the composition at a concentration ranging from about 100 μg / mL to about 1000 mg / mL, including all integers and ranges therebetween. For example, in some embodiments, one or more BT compounds are present in a concentration of about 100 μg / mL, 200 μg / mL, 300 μg / mL, 400 μg / mL, 500 μg / mL, 600 μg / mL, 700 μg / mL, 800 μg / mL, 900 μg / mL, 1000 μg / mL, 10 mg / mL, 25 mg / mL, 50 mg / mL, 100 mg / mL, 125 mg / mL, 150 mg / mL, 175 mg / mL, 200 mg / mL, 225 mg / mL, 250 mg / mL, 275 mg / mL, 300 mg / mL, 325 mg / mL, 350 mg / mL, 375 mg / mL, 380 mg / mL, 390 mg / mL, 400 mg / mL, 410 mg / mL, 420 mg / mL, 430 mg / mL, 440 mg / mL, 450 mg / mL, 460 mg / mL, 470 mg / mL, 480 mg / mL, 490 mg / mL, 500 mg / mL, 500 mg / mL, 600 mg / mL, 700 mg / mL, 800 mg / mL, 900 mg / mL, 1000 mg / mL, 10 mg / mL, 25 mg / mL, 50 mg / mL, 100 mg / mL, 125 mg / mL, 150 mg / mL, 175 mg / mL, 200 mg / mL, 225 mg / mL, 250 mg / mL, 275 mg / mL, 300 mg / mL, 325 mg / mL, 350 mg / mL, 375 mg / mL, 400 mg / mL, 450 mg In some embodiments, the one or more BT compounds are present in the composition at a concentration ranging from about 100 μg / mL to about 1000 μg / mL.

[0210] In some embodiments, the BT composition is a suspension of BisEDT in a Tween (e.g., Tween 80) in a buffer (e.g., sodium phosphate buffer). In some embodiments, the BT composition is a suspension of BisEDT in about 0.5% Tween 80 in a sodium phosphate buffer at a pH of about 7.4.

[0211] For topical application to the skin, the pharmaceutical compositions of the present disclosure may be combined with one or a combination of topical formulation carriers, which may include, but are not limited to, aqueous liquids, alcohol-based liquids, water-soluble gels, lotions, ointments, non-aqueous liquid bases, mineral oil bases, blends of mineral oil and petrolatum, lanolin, liposomes, protein carriers such as serum albumin or gelatin, powdered cellulose caramel, carbomer homopolymers, carbomer copolymers, and carbomer polymers such as Carbopol® polymers, and combinations thereof. Carbopol® polymers are, for example, polymers of acrylic acid crosslinked with polyalkenyl ethers or divinyl glycol. In some embodiments, the Carbopol® polymer is Carbopol® 71G, Carbopol® 971P, Carbopol® 974P, Carbopol® 980, Carbopol® 981, Carbopol® 5984, Carbopol® 934, Carbopol® 934P, Carbopol® 940, Carbopol® 941, and Carbopol® 1342.

[0212] Carriers for topical formulations may include semi-solid and / or gel-like vehicles that may include polymeric thickeners, water, preservatives, active surfactants, emulsifiers, and / or solvents or mixed solvent systems. US Patent No. 5,863,560 discloses a number of different carrier combinations that can aid in skin exposure to drugs, the contents of which are incorporated herein by reference. Carriers may or may not include controlled release formulations, for example, as disclosed in US2008 / 0260697, the contents of which are incorporated herein by reference. In some embodiments, carriers may or may not include viscous formulations, for example gels, as disclosed in US2009 / 0191254, the contents of which are incorporated herein by reference.

[0213] In some embodiments, the topical pharmaceutical composition of the present disclosure is provided in a sealed container. The container may be a vial, syringe, tube, bottle, ampoule (e.g., blow-fill-seal polypropylene ampoule), sachet, etc.; it may include or consist of glass, plastic, or other suitable material. For example, ampoules are generally manufactured industrially from short lengths of glass tubing formed by heating with a gas torch and gravity. Computer image recognition techniques are often employed, for example, for quality control. Filling and sealing of the ampoules may be performed by automated machines. Blank ampoules may be purchased from a scientific glass supply company and sealed, for example, with a small gas torch, preferably under an inert atmosphere. In some embodiments, the container may also be filled with an inert gas in addition to the pharmaceutical composition. In some embodiments, the composition is provided in an ampoule or other suitable container and transferred to a medium suitable for direct contact with injured skin, for example, a patch, wipe, bandage, dressing, as described below, for use.

[0214] Topical methods of delivery may include smearing (e.g., with a finger wipe or applicator), sprays, foams, bandages, sustained release patches, liquid absorbent wipes, and combinations thereof. In some embodiments, the compositions of the present disclosure are provided either directly or in a carrier(s), in a patch, wipe, bandage, dressing, or other vehicle suitable for direct contact with the skin, particularly injured skin.

[0215] In some embodiments, topical administration of the pharmaceutical composition of the present disclosure includes the use of a bandage. The pharmaceutical composition of the present disclosure may be incorporated into a bandage and / or may be applied separately with the use of a bandage. The bandage promotes healing by keeping the wound moist, creating a barrier against infection, and / or keeping the surrounding skin dry.

[0216] In some embodiments, the dressing comprises a moist wound dressing. Moist wound therapy, including the use of moist wound dressings, is the standard of care in the treatment and control of non-healing wounds, including, for example, diabetic foot ulcers. In moist wound therapy, the wound is dressed with a material that provides protection from external contaminants, prevents the wound from drying out, and provides an environment that promotes wound closure. When treating diabetic ulcers, the degree of moisture in the wound must be considered. High levels of exudate necessitate the selection of a hygroscopic material, including, but not limited to, alginates, foams, collagen-alginate combinations, carboxymethylcellulose materials, or gauze. Dry wounds with little exudate generally respond well to hydrogels. Hydrogel sheets often contain a three-dimensional network of cross-linked hydrophilic polymers. Amorphous hydrogels are similar in composition to hydrogel sheets, but lack cross-linking. The gel may also contain additional components, such as, for example, collagen, alginates, or complex carbohydrates.

[0217] The standard dressing therapy for the treatment of diabetic foot ulcers in the United States remains the use of wet-to-dry or wet-to-moist saline gauze dressings. Alginate dressings often contain calcium or calcium-sodium salts of natural polysaccharides derived from wakame seaweed. When the alginate material comes into contact with sodium-rich wound exudate, ion exchange occurs, producing a hydrophilic gel.

[0218] Additional dressing options include, but are not limited to, films, including adhesive-backed films, gels, and foams, including silicone-coated foams, hydrocolloids, collagen-based dressings, absorbent polymers, and the like. Hydrocolloid dressings often contain adhesive, absorbent, and elastomeric components. For example, carboxymethylcellulose is a common absorbent component. Hydrofiber dressings also often contain carboxymethylcellulose, e.g., sodium carboxymethylcellulose. Foam dressings often contain a polymer, often polyurethane, with small open cells that can hold fluids. Some types of foam dressings have a waterproof film covering the top surface, but may have an adhesive coating on the wound contact side or wound border. Film dressings often contain a single thin transparent sheet of polyurethane coated with an adhesive on one side. The sheet is permeable to gas and water vapor, but not to wound fluids. Hydrofiber dressings often contain sodium carboxymethylcellulose fibers. Collagen-based dressings often contain purified collagen derived from bovine, porcine, equine, or avian sources. Collagen-based dressings are thought to aid wound healing, for example, by stimulating the production of fibroblasts.

[0219] In some embodiments, the topical administration of the pharmaceutical composition of the present disclosure comprises instillation. The pharmaceutical composition of the present disclosure may be incorporated into instillation and / or may be separately applied with the use of instillation. Instillation refers to the administration of a fluid pharmaceutical composition by gradually introducing the fluid drop by drop. A typical instillation therapy instills the fluid into the wound under low positive pressure. Devices for use in instillation include, for example, Kritter-type instillation catheters (see, for example, Brent H. et al. 2005. Wounds 17(2):37-48). Techniques known in the art for improving instillation and distribution of fluids include, but are not limited to, filling the wound with instillation fluid, applying porous wound packing materials, and / or combining with negative pressure wound therapy.

[0220] In some embodiments, the topical administration of the pharmaceutical composition of the present disclosure comprises negative pressure wound therapy. Negative pressure wound therapy (NPWT) refers to the use of reduced pressure near a wound or other area of ​​injured skin to enhance and / or accelerate the growth of new tissue. The therapy involves the controlled application of less than atmospheric pressure to the area using a sealed wound dressing connected to a vacuum pump. "Negative pressure wound therapy" may also be referred to as "reduced pressure therapy" or "vacuum therapy." Typically, reduced pressure is applied to the area of ​​injured skin via a porous pad. The porous pad will contain pores that can distribute reduced pressure to the drawn area and / or channel fluid.

[0221] Many devices can be used in NPWT. NPWT devices often include a vacuum pump, a drain, and / or a dressing set. The pump may be stationary or portable, may rely on AC or battery power, and / or may allow for adjustment of suction intensity. The dressing set may include, for example, a foam or gauze dressing that is placed on the wound, and an adhesive film drape to seal the area. The drain may be supplied in a variety of configurations depending on the dressing used and the wound being treated. Once the dressing is sealed, the vacuum pump can be set to deliver continuous or intermittent pressure at levels that generally vary between -125 and -75 mmHg. For example, NPWT may be used to administer the pharmaceutical compositions of the present disclosure if the NPWT device used allows for the delivery of fluids, such as fluid pharmaceutical compositions (see, for example, Gerry R, ​​et al. 2007. Ann Plast Surg 59(1):58-62).

[0222] Using modes of administration described herein and / or known in the art, the desired dosage of the composition of the present disclosure may be delivered according to a suitable dosing regimen. Dosage and dosing regimens may vary depending on the particular formulation, route of administration, condition being treated, and other factors. Animal experiments provide reliable guidance for determining effective doses in human therapy, for example, within the skill of an ordinary physician. Interspecies scaling of effective doses may be performed by those skilled in the art according to the principles described, for example, by Mordenti, J. et al. "The use of interspecies scaling in toxicokinetics" in Toxicokinetics and New Drug Development, Yacobi et al., Eds., Pergamon Press, New York 1989, pp42-96. The pharmaceutical composition of the present disclosure can be administered according to a dosage regimen. For example, in the treatment of chronic ulcers and diabetic foot infections, including but not limited to the treatment of skin ulcers associated therewith, a first dosage regimen may be followed initially, for example, during an induction phase, and then a second dosage regimen may be followed, for example, during a maintenance phase. In some embodiments, the induction phase dosage regimen is followed for about the first 12 hours of treatment, or for about the first 18 hours, about 24 hours, about 36 hours, or about 48 hours. In some embodiments, the induction phase dosage regimen includes administration of the pharmaceutical composition of the present disclosure about every hour, about every 2 hours, about every 4 hours, about every 6 hours, for example, for the first 24 hours. In some embodiments, the induction phase dosing regimen involves administration of a pharmaceutical composition of the present disclosure once daily initially (e.g., the first 2-4 weeks), followed by a reduced dose frequency, such as twice weekly, for a subsequent period (e.g., weeks 4-12). In some embodiments, the pharmaceutical composition is administered topically according to the induction phase dosing regimen.

[0223] In some embodiments, the induction phase is followed by a maintenance phase, where, for example, a different dosing regimen may be followed. The maintenance phase may last for days, weeks, months, or longer after the initial treatment. In some embodiments, the maintenance phase lasts for about 1, 2, 3, 4, 5, 6, or 7 days after the induction phase. In some embodiments, the pharmaceutical composition is administered for 2, 3, or 4 weeks; 2, 4, 6, 8, 10, or 12 months; or 2, 3, 4, 5 years or more. In further embodiments, the pharmaceutical composition of the present disclosure is administered chronically, for example, for several years or for the patient's lifetime.

[0224] In some embodiments, the maintenance phase dosing regimen comprises administration of the pharmaceutical composition of the present disclosure at a lower dosing frequency compared to the induction phase dosing regimen. For example, in some embodiments, the pharmaceutical composition is administered about every 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, 24 hours, 30 hours, 36 hours, 42 hours, or 48 hours. In some embodiments, the maintenance phase dosing regimen comprises administration of the pharmaceutical composition of the present disclosure, for example, about every 12 hours or about every 24 hours for at least about 3 or 4 additional times following the induction phase. In some embodiments, the pharmaceutical composition is administered topically according to the maintenance phase dosing regimen. In some embodiments, the pharmaceutical composition of the maintenance phase dosing regimen is administered less frequently, for example, 1, 2, or 3 times per week, for at least about 3 or 4 additional times following the induction phase.

[0225] The preparation can be conveniently provided in unit dosage form and can be prepared by any method well known in the field of pharmacy.The amount of active ingredient that can be combined with carrier material to produce a single dosage form will vary depending on the subject to be treated, the specific mode of administration.The amount of active ingredient that can be combined with carrier material to produce a single dosage form will generally be the amount of compound that produces therapeutic effect.

[0226] The compositions can also be formulated to provide a sustained or controlled release of the active ingredient therein, for example, using hydroxypropylmethylcellulose in various proportions to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. They can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water or some other sterile injectable medium immediately prior to use.

[0227] Liquid dosage forms useful for topical administration include pharma- ceutically acceptable emulsions, lyophilisates for reconstitution, microemulsions, solutions, suspensions, gels, syrups and elixirs.In addition to the active ingredient, liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, cyclodextrin and its derivatives, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (e.g. cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof.In addition to the inert diluent, topical compositions can also contain auxiliary agents, such as wetting agents, emulsifiers and suspending agents, and preservatives.

[0228] Suspensions may contain, in addition to the active compounds, suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof.

[0229] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound may be mixed under sterile conditions with a pharma- ceutically acceptable carrier, and with any preservatives or buffers which may be required.

[0230] Ointments, pastes, creams and gels may contain, in addition to the active compound, one or more excipients or carriers, such as animal or vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc, polymers, salts, zinc oxide, or mixtures thereof. In some embodiments, the BT composition is in the form of an aqueous solution. In some embodiments, the excipient comprises a salt selected from sodium chloride or potassium chloride. In some embodiments, the excipient comprises sodium chloride.

[0231] In some cases, it is desirable to slow the absorption of a drug in order to prolong its effect. This can be accomplished by using a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends on its dissolution rate, which in turn can depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form can be accomplished by dissolving or suspending the drug in an oil vehicle.

[0232] Actual dosage levels of the active ingredients in pharmaceutical compositions can be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without being toxic to the patient.

[0233] The selected dosage level will depend upon a variety of factors, including the activity of the particular compound or combination of compounds, or esters, salts, or amides thereof, employed, the route of administration, the time of administration, the rate of excretion of the particular compound(s) employed, the duration of treatment, other drugs, compounds, and / or materials used in combination with the particular compound(s) employed, the age, sex, weight, condition, general health, and previous medical history of the subject being treated, and similar factors well known in the medical arts.

[0234] A physician or veterinarian of ordinary skill in the art can easily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, a physician or veterinarian may start the dosage of the pharmaceutical composition or compound at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved. A "therapeutically effective amount" refers to the concentration of the compound sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary depending on the subject's weight, sex, age, and medical history. Other factors that affect the effective amount can include, but are not limited to, the severity of the subject's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent administered together with the compound of the present disclosure. Even larger total doses can be delivered by multiple administrations of the drug. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher, et al. (1996), Harrison's Principles of Internal Medicine 13 ed., 1814-1882, incorporated herein by reference).

[0235] Generally, a suitable dosage of an active compound used in the compositions and methods of the present disclosure will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dosage will generally depend upon the factors described above.

[0236] The present disclosure includes the use of pharma- ceutically acceptable salts of the compounds of the present disclosure in the compositions and methods of the present disclosure. In certain embodiments, the contemplated salts of the present disclosure include, but are not limited to, alkylammonium salts, dialkylammonium salts, trialkylammonium salts, or tetraalkylammonium salts. In certain embodiments, the contemplated salts of the present disclosure include, but are not limited to, L-arginine, benentamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, the contemplated salts of the present disclosure include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts.

[0237] Pharmaceutically acceptable acid addition salts may also exist as various solvates, for example with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates may also be prepared. The source of such solvates may come from the solvent of crystallization, may be inherent in the solvent of preparation or crystallization, or may be adventitious to such solvent.

[0238] Wetting agents, emulsifying agents, and lubricating agents, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring, and perfuming agents, preservatives, and antioxidants can also be present in the composition.

[0239] Examples of pharma- ceutically acceptable antioxidants include: (1) water-soluble antioxidants such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; (2) oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, and the like; and (3) metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like. Embodiment 1. A method for treating a topical wound, comprising administering to a subject in need thereof a therapeutically effective amount of a composition comprising a bismuth thiol compound, wherein the composition is applied to an infection.

[0240] 2. The method of embodiment 1, wherein said local wound is a skin ulcer (e.g., a skin ulcer on the lower leg).

[0241] 3. The method of embodiment 2, wherein said skin ulcer is one or more of a foot ulcer, a diabetic foot ulcer, an ischemic ulcer, a gangrenous ulcer, a venous stasis ulcer, a decubitus ulcer, a Buruli ulcer, or a traumatic ulcer.

[0242] 4. The method of any one of embodiments 1-3, wherein said local wound is infected with one or more bacterial and / or fungal pathogens.

[0243] 5. The method of any of the preceding embodiments, wherein said topical wound is a diabetic foot ulcer.

[0244] 6. The method of embodiment 5, wherein said diabetic foot ulcer is a diabetic foot ulcer infection.

[0245] 7. The local wound is infected with the following bacterial pathogens: Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp., Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus. faecalis, coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium The method of any of the preceding embodiments, wherein the subject is infected with one or more of: Acinetobacter spp., S. acnes, S. dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anaerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis.

[0246] 8. The subject achieves the following outcomes after completion of dosing: a) the wound is healed or substantially healed within 12 weeks of the initial administration of the composition; and / or b) preventing amputations and / or infection-related surgery; and / or c) the wound partially or completely closes; and / or d) the wound is reduced in size from about 1% reduction compared to the original wound size to complete elimination of the wound; and / or e) the wound is 30 days or older and is healed or substantially healed;

[0247] 9. The method of embodiment 8, wherein the subject experiences two or more of the recited outcomes.

[0248] 10. The method of embodiment 8, wherein said subject experiences three or more, or four or more of said recited outcomes.

[0249] 11. The method of embodiment 8, wherein said subject experiences all of the outcomes recited above.

[0250] 12. The method of any of the preceding embodiments, wherein the BT composition comprises one or more BT compounds selected from BisBAL, BisEDT, Bis-dimercaprol, BisDTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, BisPyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, bismuth-1-mercapto-2-propanol, and Bis-EDT / 2-hydroxy-1-propanethiol.

[0251] 13. The method of embodiment 12, wherein said BT compound is BisEDT.

[0252] 14. The method of any one of embodiments 1-12, wherein the composition is a suspension of microparticles comprising the BT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

[0253] 15. The BT composition includes BisEDT, and the applied BisEDT is about 20 μg / cm 2 The method of any of the preceding embodiments, wherein the

[0254] 16. The method of any of the preceding embodiments, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to about 20 mM sodium phosphate at about pH 7.4.

[0255] 17. The method of any of the preceding embodiments, wherein after administration of the BT composition, one or more of the following occurs: (i) reducing and / or dispersing bacterial and / or fungal biofilms, (ii) impairing the growth or formation of bacterial and / or fungal biofilms, and (iii) preventing the reformation or spread of bacterial and / or fungal biofilms. 18. The method of any of the preceding embodiments, wherein the BT composition treats, manages, and / or reduces the severity of the diabetic foot infection by either or both of: (i) preventing the infection by the bacterial or fungal pathogen; and / or (ii) reducing the bacterial or fungal pathogen.

[0256] 19. The method of any of the preceding embodiments, wherein the BT composition treats, manages or reduces the severity of the infection by one or more of: (i) preventing the elaboration or secretion of exotoxins from the bacterial or fungal pathogen; (ii) inhibiting cell viability or cell proliferation of planktonic cells of the bacterial or fungal pathogen; (iii) inhibiting biofilm formation by the bacterial or fungal pathogen; (iv) inhibiting invasiveness of the biofilm or microbial pathogen into underlying tissues (e.g., subcutaneous tissue); (v) inhibiting virulence of the biofilm or microbial pathogen into underlying tissues (e.g., subcutaneous tissue); (vi) inhibiting biofilm viability or biofilm growth of biofilm-forming cells of the bacterial or fungal pathogen; and / or (vii) preventing re-formation of a biofilm following debridement.

[0257] 20. The administered BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 4. The method of any preceding embodiment, wherein the compound is present on the surface at a concentration of

[0258] 21. The administered BT composition is about 50 μg / cm 2 ~about 200μg / cm 2 The method of any of the preceding embodiments, wherein the compound is present on the surface at a concentration of

[0259] 22. The applied BT composition is about 250 μg / cm 2 ~Approx. 5,000μg / cm 2 The method of any of the preceding embodiments, wherein the compound is present on the surface at a concentration of

[0260] 23. The method of any of the preceding embodiments, wherein the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months.

[0261] 24. The method of embodiment 22, wherein the BT composition is administered once a day or three times a week.

[0262] 25. The method of any of the preceding embodiments, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

[0263] 26. The method of embodiment 25, wherein the subject is administered multiple doses of the BT composition daily or weekly for a period of about 4 weeks to about 10 weeks.

[0264] 27. The wound area is approximately 0.1 cm 2 ~about 250cm 2 4. The method of any of the preceding embodiments,

[0265] 28. A method for treating a microbial infection, comprising administering to a subject in need thereof a composition comprising a therapeutically effective amount of a bismuth thiol compound, wherein the composition is applied to the infection.

[0266] 29. The method of embodiment 28, wherein the microbial infection is a diabetic foot infection.

[0267] 30. The microbial infection is caused by the following bacterial pathogens: Staphylococcus aureus, MRSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp., Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus. faecalis, coagulase-negative Staphylococcus spp., Streptococcus spp., Corynebacterium spp., Proteus mirabilis, Bacteroides spp., Peptostreptococcus spp., Propionibacterium spp., Clostridium spp., Peptococcus spp., Prevotella spp., Finegoldia spp., Propionibacterium The method of embodiment 28 or embodiment 29, comprising one or more of the following: Acne, S. acnes, S. dysgalactiae, Serratia spp., Rhodopseudomonas spp., Bacteroides fragilis, Morganella morganii, Hemophilus spp., Enterococcus spp., Stenotrophomonas spp., Pseudomonas spp., Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp., Acinetobacter spp., Anaerococcus spp., Dialister spp., Peptoniphilus spp., Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis.

[0268] 31. The infection is associated with a wound (e.g., an ulcer) and the subject experiences the following outcomes after completion of dosing: a) the wound is healed or substantially healed within 12 weeks (e.g., within 4 weeks) of the initial administration of the composition; and / or b) preventing amputations and / or infection-related surgery; and / or c) the wound partially or completely closes; and / or d) the wound is reduced in size from about 1% reduction compared to the original wound size to complete elimination of the wound; and / or e) the wound is at least 30 days old and is healed or substantially healed;

[0269] 32. The method of embodiment 31, wherein the subject experiences two or more of the recited outcomes.

[0270] 33. The method of embodiment 31, wherein the subject experiences three or more, or four or more of the recited outcomes.

[0271] 34. The method of embodiment 31, wherein the subject experiences all of the recited outcomes.

[0272] 35. The method of any of embodiments 28-34, wherein the BT composition comprises one or more BT compounds selected from BisBAL, BisEDT, Bis-dimercaprol, BisDTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, BisPyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, bismuth-1-mercapto-2-propanol, and bis-EDT / 2-hydroxy-1-propanethiol.

[0273] 36. The method of embodiment 35, wherein the BT compound is BisEDT.

[0274] 37. The method of any one of embodiments 28 to 36, wherein the composition is a suspension of microparticles comprising the BT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

[0275] 38. The BT composition includes BisEDT, and the applied BisEDT is about 20 μg / cm 2 37. The method of embodiment 36, wherein the compound is present on the surface at a concentration greater than

[0276] 39. The method of any of embodiments 28-38, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80 (registered trademark) at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0277] 40. The method of any of embodiments 28-39, wherein the method comprises at least one of: (i) reducing and / or dispersing bacterial and / or fungal biofilms; (ii) impairing the growth or formation of bacterial and / or fungal biofilms; and (iii) preventing the reformation or spread of bacterial and / or fungal biofilms.

[0278] 41. The method of any of embodiments 28-40, wherein the BT composition treats, manages, or reduces the severity of a diabetic foot infection by one or both of: (i) preventing infection by bacterial or fungal pathogens; and (ii) reducing bacterial or fungal pathogens.

[0279] 42. The method of any of embodiments 28-41, wherein the BT composition treats, manages or reduces the severity of an infection by one or more of: (i) preventing the elaboration or secretion of exotoxins from the bacterial or fungal pathogen; (ii) inhibiting cell viability or cell proliferation of planktonic cells of the bacterial or fungal pathogen; (iii) inhibiting the formation of a biofilm or microbial pathogen by the bacterial or fungal pathogen; (iv) inhibiting the invasiveness of the biofilm into underlying tissues (e.g., subcutaneous tissue); (v) inhibiting the pathogenicity of the biofilm or microbial pathogen on underlying tissues (e.g., subcutaneous tissue); (vi) inhibiting the biofilm viability or biofilm growth of biofilm-forming cells of the bacterial or fungal pathogen; and / or (vii) preventing the reformation of a biofilm after debridement.

[0280] 43. The applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 The method of any of embodiments 28-42, wherein the compound is present on the surface at a concentration of

[0281] 44. The applied BT composition is about 50 μg / cm 2 ~Approx. 100μg / cm 2 The method of any of embodiments 28-43, wherein the compound is present on the surface at a concentration of

[0282] 45. The applied BT composition is about 250 μg / cm 2 ~Approx. 5,000μg / cm 2 The method of any of embodiments 28-43, wherein the compound is present on the surface at a concentration of

[0283] 46. ​​The method of any one of embodiments 28-45, wherein the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months.

[0284] 47. The method of embodiment 45, wherein the BT composition is administered once a day or three times a week.

[0285] 48. The method of any of embodiments 28-47, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

[0286] 49. The method of embodiment 48, wherein the subject is administered multiple doses of the BT composition daily or weekly for a period of about 10 weeks.

[0287] 50. The wound area is approximately 0.1 cm 2 ~about 250cm 2 The method of any one of embodiments 28 to 49, wherein

[0288] 51. A method for healing a wound in a subject having a diabetic foot infection, comprising administering to the subject a therapeutically effective amount of a composition comprising BisEDT, wherein the composition is a suspension of microparticles comprising BisEDT, at least 70% of the microparticles having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm, wherein the composition is applied to the infection, and wherein the wound is healed or substantially healed within 12 weeks of initial administration of the composition.

[0289] 52. The method of embodiment 51, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80 (registered trademark) at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0290] 53. The applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 The method of embodiment 51 or embodiment 52, wherein the compound is present on the surface at a concentration of

[0291] 54. The applied BT composition is about 50 μg / cm 2 ~Approx. 100μg / cm 2 The method of any of embodiments 51-53, wherein the compound is present on the surface at a concentration of

[0292] 55. The applied BT composition is about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 55. The method of any of embodiments 51-54, wherein the compound is present on the surface at a dosage of

[0293] 56. The method of any one of embodiments 51-55, wherein the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months.

[0294] 57. The method of embodiments 51-56, wherein the wound heals 4 weeks, 8 weeks, or 12 weeks after the first administration of the BT composition.

[0295] 58. The method of any of embodiments 51-57, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

[0296] 59. The method of embodiment 58, wherein the subject is administered multiple doses of the BT composition daily or weekly for a period of about 4 weeks.

[0297] 60. The wound area is approximately 0.1 cm 2 ~about 250cm 2 The method of any of embodiments 51 to 59, wherein

[0298] 61. A method for reducing the risk of amputation and / or infection-related surgery in a subject having a diabetic foot infection, comprising administering to the subject a composition comprising a therapeutically effective amount of BisEDT, wherein the composition is applied to the infection, and wherein the risk of amputation and / or infection-related surgery is reduced by about 1% to about 100% compared to a similarly situated subject not treated with a composition comprising a therapeutically effective amount of a bismuth thiol compound.

[0299] 62. The method of embodiment 61, wherein the composition is a suspension of microparticles comprising the BisEDT, and at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

[0300] 63. The method of embodiment 61 or 62, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80 (registered trademark) at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0301] 64. The applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 The method of any one of embodiments 61-63, wherein the compound is present on the surface at a concentration of

[0302] 65. The applied BT composition is about 50 μg / cm 2 ~Approx. 100μg / cm 2 The method of any of embodiments 61-64, wherein the compound is present on the surface at a concentration of

[0303] 66. The applied BT composition is about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 66. The method of any of embodiments 61-65, wherein the compound is present on the surface at a dosage of

[0304] 67. The method of any one of embodiments 61-66, wherein the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months.

[0305] 68. The method of embodiment 67, wherein the BT composition is administered once a day or three times a week.

[0306] 69. The method of any of embodiments 61-68, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

[0307] 70. The method of embodiment 69, wherein the subject is administered multiple doses of the BT composition daily or weekly for a period of about 4 weeks.

[0308] 71. Wound area is approximately 0.1 cm 2 ~about 250cm 2 The method of any of embodiments 61 to 70, wherein

[0309] 72. A method for closing a wound in a subject having a diabetic foot infection, comprising administering to the subject a composition comprising a therapeutically effective amount of BisEDT.

[0310] 73. The method of embodiment 72, wherein the composition is a suspension of microparticles comprising the BisEDT, and at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

[0311] 74. The method of embodiment 72 or 73, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80 (registered trademark) at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0312] 75. The applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 The method of any one of embodiments 72-74, wherein the compound is present on the surface at a concentration of

[0313] 76. The applied BT composition is about 50 μg / cm 2 ~Approx. 100μg / cm 2 76. The method of any of embodiments 72-75, wherein the compound is present on the surface at a concentration of

[0314] 77. The applied BT composition is about 100 μg / cm 2 Concentrations above 250 μg / cm 2 Approximately 5,000μg / cm 2 77. The method of any of embodiments 72-76, wherein the compound is present on the surface at a dosage of

[0315] 78. The method of embodiment 77, wherein the composition is applied to the infection and the wound closes within 12 weeks of the first administration of the composition.

[0316] 79. The method of embodiment 78, wherein the BT composition is administered once a day or three times a week. 80. The method of any of embodiments 72-79, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

[0317] 81. The method of embodiment 80, wherein the subject is administered multiple doses of the BT composition daily or weekly for a period of about 4 weeks.

[0318] 82. Wound area is approximately 0.1 cm 2 ~about 250cm 2 The method of any of embodiments 72 to 81, wherein

[0319] 83. A method for reducing wound size in a subject having a diabetic foot infection, comprising administering to the subject a composition comprising a therapeutically effective amount of BisEDT, wherein the composition is applied to the infection and the wound reduces in size from about 1% reduction compared to the original wound size to complete clearance of the wound within 12 weeks of initial administration of the composition.

[0320] 84. The method of embodiment 83, wherein the wound is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%.

[0321] 85. The method of embodiment 84, wherein the wound is reduced by about 50%.

[0322] 86. The method of any of embodiments 83-85, wherein the composition is a suspension of microparticles comprising the BisEDT, and at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

[0323] 87. The method of any one of embodiments 83 to 86, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80 (registered trademark) at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0324] 88. The applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 The method of any one of embodiments 83-87, wherein the compound is present on the surface at a concentration of

[0325] 89. The applied BT composition is about 50 μg / cm 2 ~Approx. 100μg / cm 2 The method of any of embodiments 83-88, wherein the compound is present on the surface at a concentration of

[0326] 90. The applied BT composition is about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 90. The method of any of embodiments 83-89, wherein the compound is present on the surface at a dosage of

[0327] 91. The method of any one of embodiments 83-90, wherein the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months.

[0328] 92. The method of embodiment 91, wherein the BT composition is administered once a day or three times a week.

[0329] 93. The method of any of embodiments 83-92, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

[0330] 94. The method of embodiment 93, wherein the subject is administered multiple doses of the BT composition daily or weekly for a period of about 4 weeks.

[0331] 95. Wound area is approximately 0.1 cm 2 ~about 250cm 2 The method of any one of embodiments 83 to 94, wherein 96. The method of any one of embodiments 83-95, wherein the wound surface area of ​​the wound is reduced by 50% 12 weeks after the first administration of the BT composition, and the BT composition is BisEDT.

[0332] 97. The method of embodiment 96, wherein the wound surface area of ​​the wound is reduced by 50% 4 weeks after the first administration of the BisEDT composition.

[0333] 98. The method of any one of embodiments 83-97, wherein the wound surface area is measured using digital photography or manual measurement.

[0334] 99. A method for preventing amputation and / or infection-related surgery in a subject having a diabetic foot infection, comprising administering to the subject a therapeutically effective amount of a BT composition.

[0335] 100. The method of embodiment 99, wherein the BT composition is a suspension of microparticles comprising the BisEDT, and at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

[0336] 101. The method of any one of embodiments 99-100, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80 (registered trademark) at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0337] 102. The applied BT composition is about 1 μg / cm 2 ~Approx. 1,000,000μg / cm 2 The method of any one of embodiments 99-101, wherein the compound is present on the surface at a concentration of

[0338] 103. The applied BT composition is about 50 μg / cm 2 ~Approx. 100μg / cm 2 The method of any of embodiments 99-102, wherein the compound is present on the surface at a concentration of

[0339] 104. The applied BT composition is about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 The method of any of embodiments 99-103, wherein the compound is present on the surface at a dosage of

[0340] 105. A method of treating a wound in a subject, wherein the wound is 30 days or older, comprising administering to the subject a therapeutically effective amount of a BT composition.

[0341] 106. The method of embodiment 105, wherein the subject has a diabetic foot infection.

[0342] 107. The method of embodiment 105 or 106, wherein the BT composition is a suspension of microparticles comprising BisEDT, and at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

[0343] 108. The method of any one of embodiments 105 to 107, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80 (registered trademark) at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to 20 mM sodium phosphate.

[0344] 109. The applied BT composition is about 1 μg / cm 2~Approx. 1,000,000μg / cm 2 The method of any one of embodiments 105-108, wherein the compound is present on the surface at a concentration of

[0345] 110. The applied BT composition is about 50 μg / cm 2 ~Approx. 100μg / cm 2 The method of any of embodiments 105-109, wherein the compound is present on the surface at a concentration of

[0346] 111. The applied BT composition is about 100 μg / cm 2 Concentrations above 250 μg / cm 2 ~Approx. 5,000μg / cm 2 The method of any of embodiments 105-110, wherein the surface is present at a dose of

[0347] 112. The method of any of embodiments 105-111, wherein the wound is more than 2 months, more than 3 months, more than 4 months, more than 5 months, more than 6 months, more than 7 months, more than 8 months, more than 9 months, more than 10 months, more than 11 months, or more than 1 year old.

[0348] 113. The method of embodiment 112, wherein the wound is more than 2 months old.

[0349] 114. The method of embodiment 112, wherein the wound is more than 3 months old.

[0350] 115. The method of any one of embodiments 1-114, wherein the BT composition comprises BisEDT, and the composition is a suspension of microparticles comprising the BisEDT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

[0351] 116. The method of embodiment 115, wherein at least 70% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm.

[0352] 117. The method of embodiment 116, wherein the VMD is from about 0.4 μm to about 3 μm.

[0353] 118. The method of embodiment 116, wherein the VMD is from about 1.0 μm to about 2.0 μm.

[0354] 119. The method of any one of embodiments 116-118, wherein at least 70%, 75%, 80%, 90%, or 95% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm.

[0355] 120. The method of embodiment 119, wherein at least 80% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm.

[0356] 121. The method of embodiment 119, wherein at least 90% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm.

[0357] 122. The method of embodiment 119, wherein substantially all of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm.

[0358] 123. The method of embodiments 119-122, wherein the VMD is from about 0.4 μm to about 3 μm.

[0359] 124. The method of any one of embodiments 119 to 122, wherein the VMD is from about 1.0 μm to about 2.0 μm.

[0360] 125. The method of any of the preceding embodiments, wherein 90% of the particles are smaller than about 10 μm.

[0361] 126. The method of embodiment 125, wherein 90% of the particles are smaller than about 10 μm, about 9 μm, about 8 μm, about 7 μm, about 6 μm, about 5 μm, about 4 μm, about 3 μm, about 3 μm, 2 μm, or about 1 μm.

[0362] 127. The method of any of the preceding embodiments, wherein the microbial species (bacteria and / or fungi) are species described in paragraphs

[0111] and / or

[0112] . EXAMPLES

[0363] The following examples are provided to illustrate the present disclosure and should not be construed as limiting thereof. Additional experimental procedures and details can be found in International Patent Application Nos. PCT / US2010 / 023108, PCT / US2011 / 023549, and PCT / US2011 / 047490, which are incorporated herein by reference in their entireties for all purposes. Example 1. General synthesis.

[0364] The starting materials and reagents used in preparing these compounds are available from commercial suppliers such as Aldrich Chemical Co., Bachem, etc., or can be prepared by methods well known in the art. The starting materials and intermediates and final products of the reactions can be isolated and purified, if desired, using conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, etc., and can be characterized using conventional means, including physical constants and spectral data. Unless otherwise noted, the reactions described herein are conducted at atmospheric pressure over a temperature range ranging from about -78°C to about 150°C.

[0365] [ka] Particulate bismuth-1,2-ethanedithiol (Bis-EDT, a soluble bismuth preparation) was prepared as follows: 0.331 L (approximately 0.575 mol) of aqueous Bi(NO3)3 (43% Bi(NO3)3 (w / w), 5% nitric acid (w / w), 52% water (w / w), Shepherd Chemical Co., Cincinnati, OH, Product No. 2362; δ approximately 1.6 g / mL) was slowly added dropwise to an excess (11.4 L) of 5% aqueous HNO3 at room temperature in a 15 L polypropylene carboy with stirring, followed by the slow addition of absolute ethanol (4 L). Some white precipitate formed but dissolved with continued stirring. A solution of 1,2-ethanedithiol (CAS540-63-6) in ethanol (approximately 1.56 L, approximately 0.55 M) was prepared separately by adding 72.19 mL (0.863 mol) of 1,2-ethanedithiol to 1.5 L of absolute ethanol using a 60 mL syringe, followed by stirring for 5 min. The 1,2-ethanedithiol / EtOH reagent was then added slowly by dropwise addition to the aqueous Bi(NO3)3 / HNO3 solution over a period of 5 h, with stirring continued overnight. The formed product was allowed to settle as a precipitate for approximately 15 min, after which the filtrate was removed using a peristaltic pump at 300 mL / min. The product was then recovered by filtration onto fine filter paper in a 15 cm diameter Buchner funnel and washed successively with three 500 mL volumes each of ethanol, USP water, and acetone to give BisEDT (694.51 gm / mole) as a yellow amorphous powder solid. The product was placed in a 500 mL amber glass bottle and dried over CaCl2 under high vacuum for 48 hours. The recovered material (yield, approximately 200 g) had a characteristic odor of thiols. The crude product was redissolved in 750 mL absolute ethanol, stirred for 30 minutes, then filtered and washed successively with 3 x 50 mL ethanol, 2 x 50 mL acetone, and again with 500 mL acetone. The rewashed powder was triturated with 1 M NaOH (500 mL), filtered, and washed with 3×220 mL water, 2×50 mL ethanol, and 1×400 mL acetone to give 156.74 gm of purified BisEDT. Subsequent batches prepared in essentially the same manner gave yields of about 78-91%.

[0366] The product is 1 H and 13 It was characterized to have the structure shown above by analyzing data from C nuclear magnetic resonance (NMR), infrared spectroscopy (IR), ultraviolet spectroscopy (UV), mass spectrometry (MS) and elemental analysis. An HPLC method was developed to measure the chemical purity of BisEDT, for which samples were prepared in DMSO (0.5 mg / mL). max was determined by scanning a solution of BisEDT in DMSO between 190 and 600 nm. Isocratic HPLC elution at 1 mL / min was performed using a YMC Pack PVC Sil NP, 5 μm, 250×4.6 mm i.d. analytical column (Waters) at 265 nm (λ max ), run at ambient temperature on a Waters (Millipore Corp., Milford, Mass.) Model 2695 chromatograph with UV detector monitoring in a mobile phase of 0.1% formic acid in acetonitrile:water (9:1) with a 2 μL injection volume, a single peak was detected reflecting a chemical purity of 100±0.1%. Elemental analysis was consistent with the structure of BisEDT as shown above.

[0367] The dried particulate matter was characterized to assess particle size characteristics. Briefly, the microparticles were resuspended in 2% Pluronic® F-68 (BASF, Mt. Olive, NJ) and the suspension was sonicated for 10 minutes in a water bath sonicator at standard settings before being analyzed using a Nanosizer / Zetasizer Nano-S particle analyzer (Model ZEN1600 (no zeta potential measurement capability), Malvern Instruments, Worcestershire, UK) according to the manufacturer's recommendations. The combined data from the two measurements showed that the microparticles exhibited a unimodal distribution with a volume mean diameter (VMD) of approximately 0.6 microns to 4 microns, with all events detectable between VMDs of approximately 0.6 microns and 4 microns, with a peak VMD at approximately 1.3 microns. Example 2. Synthesis of particulate bismuth-1,2-ethanedithiol (Bis-EDT).

[0368] Water (25.5 L) and 70% nitric acid (1800 mL) were mixed in a Nalgene reactor. Water (2300 mL) was then added to an Erlenmeyer flask followed by bismuth subnitrate (532 g) and the mixture was stirred. 70% nitric acid (750 mL) was added to the mixture to obtain a clear solution. This solution was transferred to a Nalgene reactor and the resulting mixture was stirred for 20 minutes. 9.5 L of 95% EtOH was then added to the reactor in three portions.

[0369] Separately, 1,2-ethanedithiol, 98% (229 mL) was added to the bottle, followed by two 250 mL portions of EtOH with stirring. An additional 5 L of EtOH was added to the bottle with stirring. The 1,2-ethanedithiol solution was then added to the reactor over approximately 4 hours with stirring. After stirring for 18 hours, the solids were allowed to settle for 2 hours. EtOH (20 L) was added and the mixture was stirred for 24 hours. The solids were allowed to settle for 1.5 hours and then isolated by filtration of the mixture followed by rinsing with EtOH.

[0370] The empty reactor was charged with 9 L of EtOH and the filtered solids, which was stirred for 18 hours. The solids were allowed to settle for 1 hour, then separated by filtration of the mixture, followed by rinsing with EtOH. The empty reactor was then charged with 9 L of acetone, 99.5%, and the filtered solids, which was stirred for 15 hours. The solids were allowed to settle for 1.5 hours, then separated by filtration of the mixture, followed by rinsing with acetone. Again, the empty reactor was charged with 9 L of acetone, 99.5%, and the filtered solids, which was stirred for 1.4 hours. The solids were filtered, air-dried for 69 hours, and then vacuum-dried for 4 hours. After the solids were mixed, they were sieved through a 10 mesh (2 mm) sieve, then an 18 mesh (1 mm) sieve to obtain BisEDT. Example 3. Synthesis of additional BT compounds The following bismuth thiol compounds can also be prepared according to the methods of Examples 1 and 2. [Table B]

[0371] Example 4. Establishment of a Staphylococcus aureus (S. aureus) mouse model to evaluate the efficacy of BisEDT on spleen and wound real-value index.

[0372] Establishment of the Mouse Model: A luminescent strain of Staphylococcus aureus (S. aureus) was administered intravenously to adult female Swiss Webster mice and allowed to form a biofilm over 48 hours. Mice were treated intravenously daily with 100 μL of serial dilutions of BisEDT. In this example, BisEDT was used as a suspension of drug particles (w:v) in diluent (e.g., 150, 375, or 750 μg / mL), which consisted of 3% methylcellulose / 0.5% polysorbate 80 / 10 mM sodium chloride / 10 mM sodium phosphate, pH 7.4.

[0373] Imaging studies: Mice were imaged using a Silhouette to determine wound size and volume, and bioluminescence was quantified using an In Vivo Imaging Systems (IVIS) LuminaX5.

[0374] Final colony forming units (CFU) were determined by plating homogenized tissue from the wound bed and spleen. Mice were euthanized and wound tissue and spleen were harvested. Tissue samples were weighed, placed into microfuge tubes containing sterile phosphate buffer, and homogenized. The homogenates were then serially diluted and plated to determine CFU / g tissue.

[0375] The initial study evaluated the effect of BisEDT suspension on CFU in the spleen and wound bed. The study was conducted over 11 days with 9 days of treatment. BisEDT suspension was administered to 30 mice (n=5 mice / group) at concentrations of 2500, 250, 25, 2.5, 0.25, 0.25, and 0.025 μg / mL. Comparator controls were vehicle or diluent and 50% DMSO in PBS.

[0376] Results showed that BisEDT at 2500 μg / mL reduced wound real index by approximately 5 logs compared to diluent (Figure 1) and completely inhibited splenic infection (>4-fold log reduction) compared to diluent (Figure 2).

[0377] Subsequent studies evaluated the effect of high concentrations of BisEDT suspensions on CFU in the spleen and wound bed. The study was conducted over a 12-day period with 10 days of treatment. BisEDT suspensions were administered to 20 mice (n=5 mice / group) at concentrations of 10, 5, 2.5, and 0.75 mg / mL. The negative control was untreated mice, and the positive control was mice treated with Prontosan. The results show that treatment with 10 mg / mL BisEDT reduced CFU by more than 8 logs. Complete inhibition was observed compared to the untreated control (Figure 3). Additionally, treatment with 5 mg / mL BisEDT reduced CFU by 1.5-2 logs over the Prontosan positive control. There was no significant inhibition at concentrations of BisEDT below 2.5 mg / mL (Figure 3). Treatment with 5 mg / mL or 10 mg / mL BisEDT and Prontosan completely inhibited splenic CFU (>3.5 log reduction) compared to untreated controls (Figure 4). At a dose of 2.5 mg / mL BisEDT, three mice showed no infection and one had a CFU reduction of approximately 1.5 log (Figure 4). Example 5. Mouse model to evaluate the effectiveness of BisEDT on bioluminescence The study evaluated the effect of BisEDT suspension on CFU by time-lapse bioluminescence imaging at 1, 3, 5, 7, and 9 days after treatment with BisEDT suspension. Mice were sacrificed on day 11 and the number of CFU / gram was quantified in the wound bed and spleen. Six groups of 5 mice were included in the study. Mice were treated with BisEDT at doses of 10 mg / mL, 5 mg / mL, 2.5 mg / mL, or 0.75 mg / mL. As a positive control, mice were administered Prontosan. As a negative control, mice were untreated.

[0378] Bioluminescence images of mice treated with 10 mg / mL, 5 mg / mL, 2.5 mg / mL, or 0.75 mg / mL BisEDT are contained in Figures 5A, 5B, 5C, and 5D, respectively. Bioluminescence images of mice treated with Prontosan are contained in Figure 5E, and bioluminescence images of untreated mice are contained in Figure 5F. Mice not shown did not wake up after isoflurane treatment. Mouse 1 in the 10 mg / mL BisEDT-treated group did not establish an infection with S. aureus. Mouse 4 in the 5 mg / mL BisEDT-treated group did not establish an infection with S. aureus.

[0379] Tables 1A-C show the CFU / g in the wound bed and spleen of each mouse after sacrifice. [Table 1A] [Table 1B] [Table 1C]

[0380] Example 6. A Phase 1b / 2a Randomized, Double-Blind, Placebo-Controlled, Multicenter Study to Evaluate the Efficacy of Topical BisEDT in Patients with Moderate to Severe Diabetic Foot Infections Adjunctive topical administration of BisEDT was investigated for its efficacy in resolving both infection and critical colonization, thereby removing these barriers to wound repair and closure, which represents a fundamental and clinically meaningful benefit.

[0381] Topical BisEDT suspension and placebo were used in combination with SOC systemic antibiotics (e.g., levofloxacin) for 2–3 weeks. This resulted in a similar proportion of patients in the BisEDT-treated group compared with placebo who were clinically healed at the healing visit examination, which was 2 weeks after the end of treatment. 3D photographic data of wound size suggested that BisEDT may be beneficial in promoting wound repair and closure compared with placebo. The majority of BisEDT-treated subjects had a 50% or greater reduction in wound surface area from baseline at the end of treatment (weeks 2–3), at the healing examination (weeks 4–5), and at the end of the study (weeks 8–9) (Table 2). BisEDT, when used in combination with systemic antibiotics for 4 weeks (three times per week), may be beneficial in resolving both infection and significant colonization, thus allowing wound repair and closure, a clinically meaningful baseline benefit.

[0382] [Table 2] Example 7. A Phase 1b / 2a Randomized, Double-Blind, Placebo-Controlled Study to Evaluate the Safety, Tolerability, and Efficacy of Adjunctive Therapy with Topically Applied BisEDT in Subjects With Moderate to Severe Diabetic Foot Infection (DFI) The primary objective of Part 1 of this study and the planned primary objective of Part 2 of this study was to evaluate the safety and tolerability of a locally administered BisEDT composition as an adjunct to standard of care (including systemic antibiotic therapy) in subjects with moderate to severe DFI. Secondary objectives of Part 1 of this study and planned secondary objectives of Part 2 of this study were: to evaluate the effect of the adjunct BisEDT composition on infection resolution; to evaluate the effect of the adjunct BisEDT composition on pathogen eradication; to evaluate the effect of the BisEDT composition on biofilm eradication; to evaluate the effect of the BisEDT composition on wound healing; to evaluate the effect of the BisEDT composition for prevention of lower extremity amputation; to evaluate the effect of the BisEDT composition for prevention of infection-related surgery; to evaluate the effect of the BisEDT composition on the duration of systemic antibiotic use; to evaluate the effect of the BisEDT composition on the length of hospitalization; and to evaluate the systemic absorption and pharmacokinetics of the locally administered BisEDT composition.

[0383] METHODS: This was a randomized, double-blind, placebo-controlled, multicenter study designed to be conducted in two parts. In part 1, subjects were randomized to ascending dose cohorts (outcomes of 3, 7.5, and 15 μg / cm) in a 3:1 (BisEDT vs. placebo) ratio. 2 Part 2 was planned to randomize subjects in a 1:1 ratio (BisEDT vs. placebo) based on the optimal dose identified in part 1 of the study.

[0384] Subjects with diabetes mellitus and a foot infection with an Infectious Diseases Society of America (IDSA) moderate or severe infection severity rating and a diabetic foot ulcer (DFU) present for more than 4 weeks (ensuring that the subject had a chronic persistent wound) were eligible for the study. Subjects could be enrolled if they had received antibiotic therapy within 36 hours prior to enrollment, provided there were no signs of clinical and / or microbiological failure. Subjects who required surgical treatment beyond standard bedside wound debridement or incision and drainage to treat the DFI after enrollment were not to be enrolled. Both inpatients and outpatients were eligible if they met all inclusion criteria and none of the exclusion criteria. Subjects could be screened and enrolled on 2 consecutive days.

[0385] In Part 1, subjects received either topical BisEDT or placebo applied directly to the infected ulcer three times per week for a minimum of 2 weeks (i.e., 6 doses) and a maximum of 3 weeks (i.e., 9 doses). The period between doses was no longer than 24 hours and no longer than 72 hours. The decision to discontinue topical BisEDT therapy after at least 2 weeks (i.e., 6 doses) was at the investigator's discretion and was based on resolution of clinical signs of infection.

[0386] All subjects also received systemic antibiotic treatment according to a protocol-defined algorithm, which was continued until, but not beyond, resolution of infectious findings as outlined in the 2012 IDSA Clinical Practice Guideline for the Diagnosis and Treatment of Diabetic Foot Infections.

[0387] All subjects were required to adhere to adequate pressure offloading of the foot wound (if necessary) with a removable walking orthosis from enrollment until the end of study (EOS) as directed by the treating physician.

[0388] All subjects underwent appropriate sharp debridement at baseline before the first application of BisEDT or placebo. Subjects returned for dressing changes and reapplication of BisEDT or matching placebo three times per week until cessation of treatment. The period between doses was no longer than 24 hours and no longer than 72 hours. Subjects returned to the clinic for a test of cure (TOC) visit 2 weeks after completion of treatment. An EOS visit was conducted 4 weeks after the end of treatment.

[0389] During the study, subjects underwent sharp debridement once a week, with more frequent sharp debridement occurring if medically indicated based on the investigator's clinical judgment.

[0390] A designated unblinded study nurse at each site diluted BisEDT to the designated clinical concentration, applied BisEDT or placebo to the wound, and covered the wound with one of the appropriate dressings provided. In addition, before the investigator assessed the wound for clinical signs and symptoms at each follow-up visit, the unblinded study nurse removed the previous dressing, assessed the wound for drainage, cleared any residual BisEDT or placebo via a 15-second saline irrigation of the wound, and discarded (and removed from the potential view of the blinded investigator) all old dressings and equipment before the blinded investigator assessed the wound for clinical signs and symptoms; such disposal included removal of waste disposal bags into which old dressings etc. were placed. At each visit during which application of BisEDT or placebo occurred, subject blinding was maintained by carefully limiting discussion between study staff and subjects via the use of visual barriers or subject blinding.

[0391] Treatment duration: 14-21 days; Number of subjects: 48 planned; 61 evaluated; 53 randomized; 46 completed; 7 discontinued.

[0392] Diagnosis and main criteria for inclusion: The population for this study was male and female subjects ≥18 and ≤75 years of age who had a diagnosis of diabetes; had a skin ulcer located above or distal to the ankle, was >4 weeks old, and showed clinical signs of moderate or severe infection based on IDSA guidelines; and had undergone adequate surgical debridement. Subjects should have received antibiotic therapy within 36 hours prior to enrollment, unless there were signs of clinical and / or microbiological failure and adequate arterial perfusion of the affected limb was documented.

[0393] Subjects were randomly selected from those with bone involvement (i.e., osteomyelitis), >1 co-infected DFU, or >200 cm 2 Patients were excluded from the study if they had a known or highly suspected DFU measured beyond 100 mg / kg.

[0394] Investigational and comparator information: BisEDT or placebo was applied topically to the wound. Overall levels of BisEDT dose were 3, 7.5 and 15 μg / cm 2 It was.

[0395] BisEDT was provided to the clinic as a 2.5 mg / mL suspension diluted to a specific concentration (150, 375, or 750 μg / mL) for each cohort according to the instructions for use by a designated open-label study nurse or coordinator. Suspensions of BisEDT drug particles at 150, 375, or 750 μg / mL (w:v) were prepared in a diluent composed of 3% methylcellulose / 0.5% polysorbate 80 / 10 mM sodium chloride / 10 mM sodium phosphate, pH 7.4.

[0396] The placebo (diluent) consisted of 3% methylcellulose / 0.5% polysorbate 80 / 10 mM sodium chloride / 10 mM sodium phosphate, pH 7.4 (maximum dosing volume 4 mL).

[0397] The doses administered were as follows: Cohort 1: 150 μg / mL BisEDT = 3 μg / cm 2Cohort 2: 375 μg / mL BisEDT = 7.5 μg / cm 2 Cohort 3: 750 / mL BisEDT = 15 μg / cm 2 It is.

[0398] Selection and timing of dosing for each subject: Eligible subjects were dosed according to the cohort in which they were enrolled. Subjects were dosed three times a week (e.g., Monday, Wednesday, Friday) for a minimum of two weeks and a maximum of three weeks. The period between doses was no longer than 24 hours and no longer than 72 hours.

[0399] Evaluation criteria Efficacy: The following clinical and microbiological efficacy outcomes were evaluated: Proportion of subjects who were clinically cured of infection at TOC with BisEDT or placebo 2 weeks after the end of treatment. Subjects were classified into one of the following groups: cured (resolution of all clinical signs and symptoms of infection, or sufficient improvement such that no additional antimicrobial therapy is needed), failed (insufficient resolution of at least one of the initial signs and symptoms of infection, or need for additional or alternative antimicrobial therapy, or need for surgical treatment of the infection), or indeterminate (unable to be assessed for any reason). Proportion of subjects with clinical cure at EOT with BisEDT or placebo. Proportion of subjects with confirmed or presumed microbiological cure at TOC 2 weeks after the end of treatment. Subjects were classified into one of the following groups: confirmed eradication (all original pathogens absent), presumed eradication (no material available for culture but the infection responded clinically to treatment), persistent (presence of pathogens isolated at baseline), or indeterminate (microbiological response cannot be determined). Proportion of subjects with confirmed or presumed microbiological cure at EOT and EOS. At the EOS visit, additional taxa included: colonization (isolation of a new organism not present in the initial culture, but the wound was not clinically infected), reinfection (isolation of a new organism not present in the initial culture, the wound was clinically infected), or recurrence (same organism present in the initial culture was isolated again after the intervention for confirmation or presumed microbiological response and no material to culture). Change from baseline in biofilm scores at EOT and TOC. Proportion of subjects who underwent lower limb amputation involving an infected foot. Proportion of subjects requiring infection-related surgery (i.e., lower limb amputation, incision and drainage of a new abscess, or resection of soft tissue or bone) after initiating treatment. Change from baseline in the 8-item DFI wound score at EOT, TOC, and EOS. Percentage reduction in ulcer size at EOS and each study visit.

[0400] Exploratory Efficacy: The following exploratory clinical and microbiological efficacy outcomes were evaluated: Proportion of subjects with EOS with ulcer closure (i.e., wound re-epithelialization) Length of hospital stay Duration of systemic antibiotic therapy.

[0401] Clinical Response: The distribution of clinical response outcome categories (cure, failure, and indeterminate) will be summarized descriptively by treatment group using counts and proportions at the TOC, EOT, and EOS visits.

[0402] Healing rates (healed vs. not healed) were analyzed at the TOC, EOT, and EOS visits using exact logistic regression models with covariates of treatment dose group, eight-item baseline wound score, and the interaction of treatment group with baseline wound score. If the eight-item wound score and interactions were not significant at the 10% level, they were removed from the model. If the eight-item wound score and interactions were significant at the 10% level, they were retained in the model and estimated proportions are presented at various percentiles of the eight-item baseline wound score for each treatment group. Odds ratios and 90% two-sided confidence intervals (CIs) are presented for each treatment group compared with the placebo group. Analyses are presented for both the MITT and CE populations.

[0403] Microbiological response: Distribution of microbiological response outcome categories (confirmed eradication, probable eradication, persistent, and indeterminate) will be summarized descriptively by treatment group using counts and proportions at TOC, EOT, and EOS visits. Eradication rates (confirmed + probable vs remaining) were analyzed using the same methods as clinical response. Analyses are presented for the ME population.

[0404] Colonization, Reinfection, and Relapse: At the EOS visit, the numbers and proportions of subjects with colonization, reinfection, or relapse will be tabulated by treatment group in the ME population.

[0405] Biofilm response: Distribution of absolute scores at EOT and TOC visits are summarized descriptively using counts and proportions by treatment group. Treatments were compared using the Cochran Mantel Haenszel mean score test statistic and p-values ​​are presented. Additionally, shift tables showing individual subject changes from baseline to EOT and TOC visits are presented. Analyses are presented for the ME population.

[0406] Wound score and size: The 8-item DFI wound score was calculated at baseline and each scheduled post-baseline visit. Descriptive statistics were used to summarize baseline scores, post-baseline scores, and changes from baseline by treatment group.

[0407] Changes from baseline in the 8-item DFI wound score were analyzed using repeated measures mixed effects models. Least squares (LS) means for each treatment group at each scheduled post-baseline visit are presented with 90% two-sided CIs.

[0408] The wound size component of the wound score will also be summarized separately using descriptive statistics. Percent change from baseline in wound size as assessed by investigator measurement of wound area was analyzed using the same method as the 8-item DFI wound score. Percent reduction in ulcer wound size was analyzed using ulcer wound size calculated from 3-dimensional digital photographs using the same method.

[0409] Summary of results: Subject Disposition: A total of 53 (100.0%) subjects were randomized to: 3 μg / cm 2 12 subjects in the dose group, 7.5 μg / cm 2 13 subjects in the dose group, 15 μg / cm 2 15 subjects in the dose group and 13 subjects in the placebo group. In total, 52 (98.1%) subjects received the study drug at 7.5 μg / cm 2 One person in the dose group did not receive the study drug.

[0410] Efficacy: The clinical response rate for cured subjects at the TOC visit in the MITT population was 3 μg / cm 2 66.7% of subjects in the dose group, 7.5 μg / cm 2 50.0% of subjects in the dose group, 15 μg / cm 2 The clinical response rate of cure at the EOT visit in the MITT population was 3 μg / cm 2 75.0% of subjects in the dose group, 7.5 μg / cm 2 50.0% of subjects in the dose group, 15 μg / cm 2 This was 73.3% of subjects in the dose group and 69.2% of subjects in the placebo group.

[0411] The microbiological response rate for confirmed and presumed eradication at the TOC visit in the ME population was 3 μg / cm 2 72.7% of subjects in the dose group, 7.5 μg / cm 2 60.0% of subjects in the dose group, 15 μg / cm 2 This was 45.5% of subjects in the dose group and 75.0% of subjects in the placebo group.

[0412] Colonization and reinfection rates for ME populations at EOS visit were 3 μg / cm 2 One (9.1%) subject in the dose group, 7.5 μg / cm 2 One (10.0%) subject in the dose group, 15 μg / cm 2 Recurrence occurred in two (18.2%) subjects in the dose groups and three (25.0%) subjects in the placebo group. Recurrence occurred in one (8.3%) subject in the placebo group. At the EOS visit, four (12.5%) subjects across all BisEDT dose groups experienced reinfection or recurrence compared with four (33.3%) subjects in the placebo group, a reduction of approximately one-third with BisEDT.

[0413] In total, a baseline biofilm score of 2 or greater (suggesting the presence of biofilm) for the ME population was reported in 27 (61.4%) subjects: 3 μg / cm 2 Nine subjects (81.8%) in the dose group were 7.5 μg / cm 2 Six subjects (60.0%) in the dose group were 15 μg / cm 2 Five subjects (45.5%) in the dose group and seven subjects (58.3%) in the placebo group.

[0414] At the EOT visit in the ME population, 13 (43.3%) subjects across all dose groups combined and 7 (58.3%) subjects in the placebo group reported a biofilm score of ≥2.At the TOC visit in the ME population, 17 (56.7%) subjects across all dose groups combined and 6 (50.0%) subjects in the placebo group reported a biofilm score of ≥2.

[0415] The mean change in the 8-item DFI wound score from baseline to the EOT and TOC visits for the MITT population was 3 μg / cm 2 The mean values ​​for the 7.5 μg / cm 2 The mean values ​​were −6.7 and −8.0 in the 15 μg / cm 2 The mean mean scores for the dose groups were -10.2 and -10.4, respectively, and for the placebo group they were -8.9 and -9.0, respectively.

[0416] The median reduction from baseline in ulcer wound size based on 3-dimensional photographs at the EOT, TOC, and EOS visits for the MITT population was 3 μg / cm 2 32.8%, 71.3%, and 97.4% in the 7.5 μg / cm dose groups, respectively. 2 62.4%, 88.6%, and 86.2% in the 15 μg / cm dose group, respectively. 2 The median reductions in ulcer wound size from baseline based on 3D photographs at the EOT, TOC, and EOS visits were 41.5%, 78.2%, and 85.2%, respectively, in the MITT population for all dose groups combined, and 6.1%, 26.5%, and 29.7%, respectively, in the placebo group, an approximately three-fold increase compared with placebo.

[0417] The number and percentage of subjects who achieved a >50% reduction in wound size based on 3D photography at the TOC visit were >3 μg / cm 2 Six subjects (54.5%) in the dose group were 7.5 μg / cm 2 Six subjects (66.7%) in the dose group were 15 μg / cm 2 There were eight subjects (57.1 percent) in the dose group and five subjects (38.5 percent) in the placebo group. The number and percentage of subjects who had a >50% reduction in wound size based on three-dimensional photographs at the EOS visit were >3 μg / cm 2 Six subjects (54.5%) in the dose group were 7.5 μg / cm 2 Five subjects (62.5%) in the dose group received 15 μg / cm 2There were 9 (64.3 percent) subjects in the dose group and 5 (41.7 percent) subjects in the placebo group. The number and proportion of subjects who achieved a >50% reduction in wound size based on 3-D photographs at the TOC and EOS visits was 20 (58.8%) and 20 (60.6%), respectively, in the MITT population for all dose groups combined, and 5 (38.5%) and 5 (41.7%), respectively, in the placebo group, a roughly 1.5-fold increase with BisEDT compared with placebo.

[0418] A total of two (15.4%) placebo subjects underwent amputation related to a target infected ulcer compared with one (2.6%) of all BisEDT-treated subjects, which was reduced by approximately sixfold with BisEDT.

[0419] Conclusions: This study demonstrated that BisEDT was safe and well tolerated with minimal systemic exposure.

[0420] Part 1 of the study was not designed to demonstrate the efficacy of BisEDT as an adjunct to standard of care based on the small sample size. As part of standard of care, all subjects in the safety population of this study received BisEDT (7.5 μg / cm 2 All subjects in the 10-dose group (except for one subject in the 10-dose group) received systemic antibiotic therapy to treat DFI. Based on the limited data collected, the following observations can be made regarding BisEDT (adjunct to standard care) and placebo (standard care alone):

[0421] The data showed that 4 (12.5%) subjects across all dose groups experienced reinfection or recurrence, indicating that BisEDT may be beneficial in preventing reinfection and recurrence based on the effect seen at the EOS visit, which was reduced by approximately one-third with BisEDT compared to 4 (33.3%) subjects in the placebo group.

[0422] The data showed that BisEDT may be beneficial in preventing target ulcer-specific amputations and target ulcer-specific infection-related surgical interventions when compared with placebo. A total of two (15.4%) placebo subjects underwent amputations related to a target infected ulcer compared with one (2.6%) of all BisEDT-treated subjects, which was approximately six-fold reduced with BisEDT.

[0423] The data indicated that BisEDT may be beneficial in reducing biofilm, as a biofilm score of ≥2 was reported at the EOT visit in 13 (43.3%) subjects combined across all dose groups and 7 (58.3%) subjects in the placebo group.

[0424] The data indicated that BisEDT may be beneficial as an adjunct to wound healing in subjects with moderate DFI based on the effects seen in the percentage of ulcer size reduction and the proportion of subjects with a greater than 50% reduction in wound size. The proportion of subjects with a greater than 50% reduction in wound size based on 3D photographs at the TOC and EOS visits was 58.8% and 60.6% in the MITT population combined for all dose groups, respectively, and 38.5% and 41.7% in the placebo group, respectively, an approximately 1.5-fold increase with BisEDT compared to placebo. The median percentage reduction in ulcer wound size from baseline based on 3D photographs at the TOC and EOS visits was 78.2% and 85.2% in the MITT population combined for all dose groups, respectively, and 26.5% and 29.7% in the placebo group, respectively, an approximately 3-fold increase compared to placebo.

[0425] BisEDT may be beneficial in preventing target ulcer-specific amputations and target ulcer-specific infection-related surgical interventions when compared with placebo.

[0426] Furthermore, BisEDT may be beneficial in preventing reinfection and recurrence: the EOS visit showed that 4 (12.5%) subjects in all dose groups combined experienced reinfection or recurrence, compared with 4 (33.3%) subjects in the placebo group.

[0427] Furthermore, BisEDT may be beneficial in reducing biofilms as demonstrated at the EOT visit, where a biofilm score of ≥2 was reported in 13 (43.3%) subjects across all dose groups combined and 7 (58.3%) subjects in the placebo group.

[0428] BisEDT may also be beneficial as an adjunctive wound healing treatment in subjects with moderate DFI, based on the effects seen in subjects who experienced a >50% reduction in wound size. The median reduction in ulcer wound size from baseline based on 3D photography at the TOC and EOS visits was 78.2% and 85.2%, and 26.5% and 29.7, respectively, in the MITT population across all dose groups combined.

[0429] Based on the limited whole blood concentration data collected, BisEDT does not appear to present relevant systemic exposure.

[0430] Wound Measurements (by Investigator): Size (cm) 2 ):The approximate area of ​​the DFU was calculated based on the maximum length and width of the ulcer. Depth (mm): Depth was measured as the deepest apparent part of the wound, using a sterile cotton-tipped swab held at 90 degrees to the wound and marked with a pen held parallel to the unwounded skin surface. Subcutaneous tissue stripping (mm): Any tunneling, subepithelial tissue loss, or shearing was measured using a sterile cotton-tipped swab and marked with a pen held perpendicular to the swab.

[0431] Microbiological evaluation. Cultures were collected at baseline and as needed for the subject's treatment. Tissue samples were sent to a central laboratory where the presence of viable microorganisms was determined by standard microbiological culture methods and included species analysis. Antibiotic susceptibility testing was performed automatically for important pathogens for which antimicrobial resistance may affect treatment. Examples of important pathogens included MRSA, methicillin-susceptible Staphylococcus aureus (MSSA), beta-hemolytic streptococci, enterococci, Enterobacteriaceae, P aeruginosa, and other non-fermenting gram-negative bacilli. Antibiotic susceptibility testing for organisms not on the list of important pathogens may also have been performed at the request of the investigator or sponsor. Susceptibility of bacterial isolates to BisEDT was also determined but was not reported to the investigator.

[0432] Efficacy variables and assessments.

[0433] Efficacy Outcomes. The following clinical and microbiological efficacy outcomes were evaluated: Proportion of subjects who were clinically cured of infection at TOC with BisEDT or placebo 2 weeks after the end of treatment. Subjects were classified into one of the following groups: cured (resolution of all clinical signs and symptoms of infection, or sufficient improvement such that no additional antimicrobial therapy is needed), failed (insufficient resolution of at least one of the initial signs and symptoms of infection, or need for additional or alternative antimicrobial therapy, or need for surgical treatment of the infection), or indeterminate (unable to be assessed for any reason). Proportion of subjects with clinical cure at EOT with BisEDT or placebo. Proportion of subjects with confirmed or presumed microbiological cure at TOC 2 weeks after the end of treatment. Subjects were classified into one of the following groups: confirmed eradication (all original pathogens absent), presumed eradication (no material available for culture but the infection responded clinically to treatment), persistent (presence of pathogens isolated at baseline), or indeterminate (microbiological response cannot be determined). Proportion of subjects with confirmed or presumed microbiological cure at EOT and EOS. At the EOS visit, additional taxa included: colonization (isolation of a new organism not present in the initial culture, but the wound was not clinically infected), reinfection (isolation of a new organism not present in the initial culture, the wound was clinically infected), or recurrence (same organism present in the initial culture was isolated again after the intervention for confirmation or presumed microbiological response and no material to culture). Change from baseline in biofilm scores at EOT and TOC. Proportion of subjects who underwent lower limb amputation involving an infected foot. Proportion of subjects requiring infection-related surgery (i.e., lower limb amputation, incision and drainage of a new abscess, or resection of soft tissue or bone) after initiating treatment. Change from baseline in the 8-item DFI wound score at EOT, TOC, and EOS. Percentage reduction in ulcer size at EOS and each study visit.

[0434] Exploratory Outcomes: The following exploratory efficacy outcomes were assessed: Proportion of subjects with EOS with ulcer closure (i.e., wound re-epithelialization) Length of hospital stay Duration of systemic antibiotic therapy Biofilm response: the following categories: score 0, no bacteria observed; 1, single cell; 2, small microcolonies (approximately 10 cells); 3, large microcolonies (approximately 100 cells); 4, continuous film; 5, thick continuous film was used. Subjects with no debridement material as a result of DFI healing were given a score of 0. Distribution of absolute scores at EOT and TOC visits is summarized descriptively using counts and percentages by treatment group. Treatments are compared using the Cochran Mantel Haenszel mean score test statistic and p values ​​are presented. Additionally, shift tables showing individual subject changes from baseline to EOT and TOC visits are presented. Analyses are presented for the ME population.

[0435] Wound score and size: The 8-item DFI wound score was calculated at baseline and at each scheduled post-baseline visit. Descriptive statistics were used to summarize baseline scores, post-baseline scores, and change from baseline by treatment group. Change from baseline in the 8-item DFI wound score was analyzed using a repeated measures mixed effects model. Change from baseline in score was the response variable in the model with treatment group, time point, and treatment group by time point interaction as factors. Least squares (LS) means for each treatment group at each scheduled post-baseline visit are presented with 90% two-sided CIs. The wound size component of the wound score is also summarized separately using descriptive statistics. Percent change from baseline in wound size, as assessed by investigator measurements of wound area, was analyzed using the same methods as the 8-item DFI wound score. Percent reduction in ulcer wound size was analyzed using ulcer wound size calculated from 3-dimensional digital photographs using the same methods. Analyses are presented for both the MITT and CE populations.

[0436] Systemic antibiotic therapy: Table 3 summarizes concomitant antibiotics for the MITT population. The most common concomitant antibiotic was levofloxacin (49 [94.2%] subjects: 3 μg / cm 2 Twelve [100.0%] subjects in the dose group, 7.5 μg / cm 2 Ten [83.3 percent] subjects in the dose group, 15 μg / cm 2 14 [93.3 subjects] in the dose group and 13 [100.0%] subjects in the placebo group); this was also the most common antibiotic used to treat DFI. [Table 3-1] [Table 3-2]

[0437] Concomitant medications: In the MITT cohort, all 52 (100.0%) subjects were taking at least one concomitant medication. The most commonly reported non-antibiotic concomitant medications were: Insulin glargine: 3 μg / cm 2 Seven (58.3%) subjects in the dose group were 7.5 μg / cm 2 Four (33.3%) subjects in the dose group, 15 μg / cm 2 Eight (53.3%) subjects in the dose group and nine (69.2%) subjects in the placebo group Lisinopril: 3 μg / cm 2 Seven (58.3%) subjects in the dose group were 7.5 μg / cm 2 Four (33.3%) subjects in the dose group, 15 μg / cm 2 6 (40.0%) subjects in the dose group and 6 (46.2%) subjects in the placebo group Metformin: 3 μg / cm 2 Three subjects (25.0%) in the dose group were 7.5 μg / cm 2 Five subjects (41.7%) in the dose group were 15 μg / cm 2 Seven subjects (46.7%) in the dose group and two subjects (15.4%) in the placebo group Treatment Compliance: Overall, 3 (5.8%) subjects missed one dose of medication (3 μg / cm 2 Two subjects (16.7%) in the dose group and one subject (7.7%) in the placebo group missed two doses (3 μg / cm 2 One (8.3%) subject in the 1000 mg / kg dose group and one (1.9%) subject missed three doses (3 μg / cm 2 one [8.3%] subject in the dose group).

[0438] Efficacy and Pharmacokinetic Results: Clinical Response: Clinical response at the test of cure visit. Table 4 presents an analysis of clinical response at the TOC visit for the MITT population. No differences were observed between treatment groups. [Table 4]

[0439] Clinical Response at End of Treatment Visit: Table 5 presents an analysis of clinical response at the EOT visit for the MITT population. No differences were observed between treatment groups. [Table 5]

[0440] Microbiological Response: Microbiological response at test of cure visit. Table 6 presents an analysis of the microbiological response at the TOC visit for the ME population. No differences were observed between treatment groups. [Table 6]

[0441] Exploratory Results: Microbiological Response at End of Treatment Visit. Table 7 presents the analysis of microbiological response at the EOT visit for the ME population. No differences were observed between treatment groups. [Table 7]

[0442] Microbiological response at the End of Study Visit: Table 8 presents an analysis of the microbiological response at the EOS visit for the ME population. No differences were observed between treatment groups. [Table 8]

[0443] Table 9 summarizes colonization, reinfection, and recurrence at EOS visit for the ME population. No differences were observed between treatment groups. Four subjects (12.5%) across all dose groups combined experienced reinfection or recurrence, a reduction of approximately one-third with BisEDT compared to four subjects (33.3%) in the placebo group. [Table 9]

[0444] Biofilm scores. Biofilm scores at the test of cure visit: Table 10 presents an analysis of the biofilm response at the TOC visit for the ME population. In total, 27 (61.4%) subjects had a baseline biofilm score of 2 or greater (indicating the presence of biofilm): 2 Nine (81.8%) subjects in the dose group were 7.5 μg / cm 2 Six (60.0%) subjects in the dose group, 15 μg / cm 2 Biofilm scores of ≥2 were reported in 17 (56.7%) subjects across all dose groups and 6 (50.0%) subjects in the placebo group at the TOC visit in the ME population. No differences in biofilm responses at the TOC visit were observed between treatment groups in the ME population.

[0445] [Table 10] Biofilm scores at end of treatment visit: Table 11 presents an analysis of biofilm response at the EOT visit for the ME population. A biofilm score of 2 or greater at the EOT visit (indicating the presence of a biofilm) was reported in 13 (43.3%) subjects across all dose groups combined in the ME population and 7 (58.3%) subjects in the placebo group. No differences in biofilm response at the EOT visit were observed between treatment groups in the ME population. [Table 11]

[0446] Lower Limb Amputations: The number and proportion of subjects with lower limb amputations involving the infected foot in the MITT population was >3 μg / cm 2 0 (0.0%) subjects in the dose group, 7.5 μg / cm 2One (8.3%) subject in the 15 μg / cm 2 There was one (6.7%) subject in the 7.5 μg / cm dose group and two (15.4%) subjects in the placebo group. 2 In the 15 μg / cm dose group, subject 001-026 had an amputation associated with a new infection of a non-target ulcer on the forefoot. 2 In the dose group, subject 001-031 had an amputation related to worsening target ulcer infection. In the placebo group, subject 001-013 had an amputation related to a new diagnosis of osteomyelitis in a target ulcer on the right foot. In the placebo group, subject 006-001 had an amputation related to worsening osteomyelitis in a target ulcer on the left foot. In all dose groups combined, 1 of 39 (2.56%) subjects had a lower extremity amputation related to a target infected ulcer compared with 2 of 13 (15.4%) subjects in the placebo group.

[0447] Infection-related surgery: The number and proportion of subjects undergoing infection-related surgical intervention in the MITT population was >3 μg / cm 2 0 (0.0%) subjects in the dose group, 7.5 μg / cm 2 One (8.3%) subject in the dose group, 15 μg / cm 2 one (6.7%) subject in the dose group and two (15.4%) subjects in the placebo group.

[0448] 8-item DFI Wound Score: Table 12 summarizes the 8-item DFI Wound Score by visit and change from baseline for the MITT population. [Table 12]

[0449] Ulcer Size: Table 13 presents a summary of ulcer wound size and percent change from baseline based on...

Claims

1. 1. A method for treating a topical wound, comprising administering to a subject in need thereof a composition comprising a therapeutically effective amount of a bismuth thiol compound, wherein the composition is applied to an infection.

2. 2. The method of claim 1, wherein the localized wound is a skin ulcer (e.g., a skin ulcer on the lower leg).

3. 3. The method of claim 2, wherein the skin ulcer is one or more of a foot ulcer, a diabetic foot ulcer, an ischemic ulcer, a gangrenous ulcer, a venous stasis ulcer, a decubitus ulcer, a Buruli ulcer, or a traumatic ulcer.

4. The method of any one of claims 1 to 3, wherein the topical wound is infected with one or more bacterial and / or fungal pathogens.

5. 10. The method of any preceding claim, wherein the topical wound is a diabetic foot ulcer.

6. 6. The method of claim 5, wherein the diabetic foot ulcer is a diabetic foot ulcer infection.

7. The topical wound is free of the bacterial pathogens Staphylococcus aureus, MRSA, MSSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp., Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, Coagulase-negative Staphylococcus spp., Streptococcus spp. , Corynebacterium spp. , Proteus mirabilis, Bacteroides spp. , Peptostreptococcus spp. , Propionibacterium spp. , Clostridium spp. , Peptococcus spp. , Prevotella spp. , Finegoldia spp. , Propionibacterium acnes, S. dysgalactiae, Serratia spp. , Rhodopseudomonas spp. , Bacteroides fragilis, Morganella morganii, Hemophilus spp. , Enterococcus spp. , Stenotrophomonas spp. , Pseudomonas spp. , Stenotrophomonas maltophilia, Enterobacter Cloacae, Sphingomonas sp. , Acinetobacter spp. , Anerococcus spp. , Dialister spp. , Peptoniphilus spp. , Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, Anaerococcus vaginalis.

8. The subject achieves the following outcomes after completion of dosing: f) the wound is healed or substantially healed within 12 weeks of the initial administration of the composition; and / or g) preventing amputations and / or infection-related surgery; and / or h) the wound partially or completely closes; and / or i) the wound is reduced in size from about 1% reduction compared to the original wound size to complete elimination of the wound; and / or j) the wound is 30 days or older and is healed or substantially healed.

9. 10. The method of claim 8, wherein the subject experiences two or more of the recited outcomes.

10. 9. The method of claim 8, wherein the subject experiences three or more, or four or more of the recited outcomes.

11. 10. The method of claim 8, wherein the subject experiences all of the recited outcomes.

12. 13. The method of any of the preceding claims, wherein the BT composition comprises one or more BT compounds selected from BisBAL, BisEDT, Bis-dimercaprol, BisDTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, BisPyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, Bismuth-1-mercapto-2-propanol, and Bis-EDT / 2-hydroxy-1-propanethiol.

13. The method of claim 12, wherein the BT compound is BisEDT.

14. The method according to any one of claims 1 to 12, wherein the composition is a suspension of microparticles comprising the BT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

15. The BT composition includes BisEDT, and the applied BisEDT is about 20 μg / cm ２ 2. The method of claim 1, wherein the at least one of the first and second electrodes is present on the surface in a concentration of more than 100 μg / cm.

16. 2. The method of any of the preceding claims, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05 to 40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2 to about 20 mM sodium phosphate.

17. 13. The method according to any of the preceding claims, wherein after administration of the BT composition, one or more of the following occurs: (i) reducing and / or dispersing bacterial and / or fungal biofilms, (ii) impairing the growth or formation of bacterial and / or fungal biofilms, and (iii) preventing the reformation or spread of bacterial and / or fungal biofilms.

18. 13. The method according to any of the preceding claims, wherein the BT composition treats, manages and / or reduces the severity of the diabetic foot infection by either or both of: (i) preventing the infection by the bacterial or fungal pathogens; and / or (ii) reducing the bacterial or fungal pathogens.

19. 5. The method of any of the preceding claims, wherein the BT composition treats, manages or reduces the severity of an infection by one or more of: (i) preventing the elaboration or secretion of exotoxins from the bacterial or fungal pathogen; (ii) inhibiting cell viability or cell proliferation of planktonic cells of the bacterial or fungal pathogen; (iii) inhibiting biofilm formation by the bacterial or fungal pathogen; (iv) inhibiting the invasiveness of the biofilm or microbial pathogen into underlying tissues (e.g. subcutaneous tissue); (v) inhibiting the virulence of the biofilm or microbial pathogen into underlying tissues (e.g. subcutaneous tissue); (vi) inhibiting biofilm viability or biofilm growth of biofilm forming cells of the bacterial or fungal pathogen; and / or (vii) preventing biofilm reformation after debridement.

20. The administered BT composition is about 1 μg / cm ２ ~Approx. 1,000,000μg / cm ２ 2. The method of claim 1, wherein the compound is present on the surface in a concentration of

21. The administered BT composition is about 50 μg / cm ２ ~Approx. 200μg / cm ２ 2. The method of claim 1, wherein the compound is present on the surface in a concentration of

22. The applied BT composition is about 250 μg / cm ２ ~Approx. 5,000μg / cm ２ 2. The method of claim 1, wherein the compound is present on the surface in a concentration of

23. 13. The method according to any of the preceding claims, wherein the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months.

24. 23. The method of claim 22, wherein the BT composition is administered once a day or three times a week.

25. 13. The method according to any of the preceding claims, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

26. 26. The method of claim 25, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of about 4 weeks to about 10 weeks.

27. The wound area is approximately 0.1 cm ２ ~Approx. 250cm ２ 2. A method according to any preceding claim, wherein:

28. 1. A method for treating a microbial infection comprising administering to a subject in need thereof a composition comprising a therapeutically effective amount of a bismuth thiol compound, wherein the composition is applied to the infection.

29. 29. The method of claim 28, wherein the microbial infection is a diabetic foot infection.

30. The microbial infection may be caused by any of the following bacterial pathogens: Staphylococcus aureus, MRSA, MSSA, Escherichia coli, Pseudomonas aeruginosa, Citrobacter spp., Klebsiella oxytoca, Proteus spp., Mobiluncus spp., Gardenella spp., Atopibium spp., S. epidermidis, Enterococcus faecalis, Coagulase-negative Staphylococcus spp., Streptococcus spp. , Corynebacterium spp. , Proteus mirabilis, Bacteroides spp. , Peptostreptococcus spp. , Propionibacterium spp. , Clostridium spp. , Peptococcus spp. , Prevotella spp. , Finegoldia spp. , Propionibacterium acnes, S. dysgalactiae, Serratia spp. , Rhodopseudomonas spp. , Bacteroides fragilis, Morganella morganii, Hemophilus spp. , Enterococcus spp. , Stenotrophomonas spp. , Pseudomonas spp. , Stenotrophomonas maltophilia, Enterobacter cloacae, Sphingomonas sp. , Acinetobacter spp. , Anerococcus spp. , Dialister spp. , Peptoniphilus spp.

30. The method of claim 28 or 29, comprising one or more of the following: Finegoldia magna, Peptoniphilus asaccharolyticus, Veillonella atypia, and Anaerococcus vaginalis.

31. The infection is associated with a wound (e.g., an ulcer) and the subject experiences the following outcomes after completion of dosing: a) the wound is healed or substantially healed within 12 weeks (e.g., within 4 weeks) of the initial administration of the composition; and / or b) preventing amputations and / or infection-related surgery; and / or c) the wound partially or completely closes; and / or d) the wound is reduced in size from about 1% reduction compared to the original wound size to complete elimination of the wound; and / or e) the wound is 30 days or older and is healed or substantially healed.

32. 32. The method of claim 31 , wherein the subject experiences two or more of the recited outcomes.

33. 32. The method of claim 31 , wherein the subject experiences three or more, or four or more of the recited outcomes.

34. 32. The method of claim 31 , wherein the subject experiences all of the recited outcomes.

35. 35. The method of any of claims 28 to 34, wherein the BT composition comprises one or more BT compounds selected from BisBAL, BisEDT, Bis-dimercaprol, BisDTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, BisPyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, Bismuth-1-mercapto-2-propanol, and Bis-EDT / 2-hydroxy-1-propanethiol.

36. 36. The method of claim 35, wherein the BT compound is BisEDT.

37. The method of any one of claims 28 to 36, wherein the composition is a suspension of microparticles comprising the BT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

38. The BT composition includes BisEDT, and the applied BisEDT is about 20 μg / cm ２ 37. The method of claim 36, wherein the at least one of the first and second electrodes is present on the surface at a concentration greater than about 1000 μg / cm.

39. 39. The method of any of claims 28-38, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05-40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2-20 mM sodium phosphate.

40. 40. The method of any of claims 28 to 39, wherein the method comprises at least one of: (i) reducing and / or dispersing bacterial and / or fungal biofilms; (ii) impairing the growth or formation of bacterial and / or fungal biofilms; and (iii) preventing the reformation or spread of bacterial and / or fungal biofilms.

41. The method according to any of claims 28 to 40, wherein the BT composition treats, manages or reduces the severity of the diabetic foot infection by one or both of: (i) preventing the infection by the bacterial or fungal pathogen; and (ii) reducing the bacterial or fungal pathogen.

42. 42. The method of any of claims 28 to 41, wherein the BT composition treats, manages or reduces the severity of the infection by one or more of: (i) preventing the elaboration or secretion of exotoxins from the bacterial or fungal pathogen; (ii) inhibiting cell viability or cell proliferation of planktonic cells of the bacterial or fungal pathogen; (iii) inhibiting biofilm or microbial pathogen formation by the bacterial or fungal pathogen; (iv) inhibiting biofilm invasiveness into underlying tissues (e.g. subcutaneous tissue); (v) inhibiting virulence of biofilm or microbial pathogens towards underlying tissues (e.g. subcutaneous tissue); (vi) inhibiting biofilm viability or biofilm growth of biofilm forming cells of the bacterial or fungal pathogen; and / or (vii) preventing biofilm reformation after debridement.

43. The applied BT composition is about 1 μg / cm ２ ~Approx. 1,000,000μg / cm ２ The method according to any one of claims 28 to 42, wherein the compound is present on the surface at a concentration of

44. The applied BT composition is about 50 μg / cm ２ ~Approx. 100μg / cm ２ The method of any of claims 28 to 43, wherein the compound is present on the surface at a concentration of

45. The applied BT composition is about 250 μg / cm ２ ~Approx. 5,000μg / cm ２ The method of any of claims 28 to 43, wherein the compound is present on the surface at a concentration of

46. 46. ​​The method of any one of claims 28 to 45, wherein the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months.

47. 46. ​​The method of claim 45, wherein the BT composition is administered once a day or three times a week.

48. 48. The method according to any of claims 28-47, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

49. 49. The method of claim 48, wherein the subject is administered multiple doses of the BT composition daily or weekly for a period of about 10 weeks.

50. The wound area is approximately 0.1 cm ２ ~Approx. 250cm ２ The method according to any one of claims 28 to 49, wherein

51. 1. A method of healing a wound in a subject having a diabetic foot infection, comprising administering to the subject a therapeutically effective amount of a composition comprising BisEDT, said composition being a suspension of microparticles comprising BisEDT, at least 70% of said microparticles having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm, said composition being applied to the infection, and wherein the wound is healed or substantially healed within 12 weeks of initial administration of the composition.

52. 52. The method of claim 51, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05-40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2-20 mM sodium phosphate.

53. The applied BT composition is about 1 μg / cm ２ ~Approx. 1,000,000μg / cm ２ 53. The method of claim 51 or claim 52, wherein the compound is present on the surface at a concentration of

54. The applied BT composition is about 50 μg / cm ２ ~Approx. 100μg / cm ２ The method of any of claims 51 to 53, wherein the compound is present on the surface at a concentration of

55. The applied BT composition is about 100 μg / cm ２ Concentrations greater than about 250 μg / cm ２ ~Approx. 5,000μg / cm ２ 55. The method of any one of claims 51 to 54, wherein the compound is present on the surface in a dosage of

56. 56. The method of any one of claims 51 to 55, wherein the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months.

57. The method according to claims 51 to 56, wherein the wound is healed 4 weeks, 8 weeks, or 12 weeks after the first administration of the BT composition.

58. 58. The method according to any of claims 51-57, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

59. 59. The method of claim 58, wherein the subject is administered multiple doses of the BT composition daily or weekly for a period of about 4 weeks.

60. The wound area is approximately 0.1 cm ２ ~Approx. 250cm ２ The method according to any one of claims 51 to 59, wherein

61. 1. A method for reducing the risk of amputation and / or infection-related surgery in a subject having a diabetic foot infection, comprising administering to the subject a composition comprising a therapeutically effective amount of BisEDT, wherein the composition is applied to the infection, and wherein the risk of amputation and / or infection-related surgery is reduced by about 1% to about 100% compared to a similarly situated subject not being treated with a composition comprising a therapeutically effective amount of a bismuth thiol compound.

62. 62. The method of claim 61, wherein the composition is a suspension of microparticles comprising the BisEDT, and at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

63. 63. The method of claim 61 or 62, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05-40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2-20 mM sodium phosphate.

64. The applied BT composition is about 1 μg / cm ２ ~Approx. 1,000,000μg / cm ２ The method of claims 61 to 63, wherein the compound is present on the surface at a concentration of

65. The applied BT composition is about 50 μg / cm ２ ~Approx. 100μg / cm ２ The method of any one of claims 61 to 64, wherein the compound is present on the surface at a concentration of

66. The applied BT composition is about 100 μg / cm ２ Concentrations greater than about 250 μg / cm ２ ~Approx. 5,000μg / cm ２ 66. The method of any one of claims 61 to 65, wherein the compound is present on the surface in a dosage of

67. 67. The method of any one of claims 61 to 66, wherein the BT composition is administered three times a day, twice a day, once a day, every other day, once every third day, three times a week, once a week, once every two weeks, once a month, or once every two months.

68. 68. The method of claim 67, wherein the BT composition is administered once a day or three times a week.

69. 69. The method according to any of claims 61-68, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

70. 70. The method of claim 69, wherein the subject is administered multiple doses of the BT composition daily or weekly for a period of about 4 weeks.

71. The wound area is approximately 0.1 cm ２ ~Approx. 250cm ２ The method according to any one of claims 61 to 70, wherein

72. 11. A method of closing a wound in a subject having a diabetic foot infection, comprising administering to the subject a composition comprising a therapeutically effective amount of BisEDT.

73. 73. The method of claim 72, wherein the composition is a suspension of microparticles comprising the BisEDT, at least 70% of the microparticles having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

74. 74. The method of claim 72 or 73, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05-40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2-20 mM sodium phosphate.

75. The applied BT composition is about 1 μg / cm ２ ~Approx. 1,000,000μg / cm ２ The method of any one of claims 72 to 74, wherein the compound is present on the surface at a concentration of

76. The applied BT composition is about 50 μg / cm ２ ~Approx. 100μg / cm ２ The method of any of claims 72 to 75, wherein the compound is present on the surface at a concentration of

77. The applied BT composition is about 100 μg / cm ２ Concentrations greater than about 250 μg / cm ２ ~Approx. 5,000μg / cm ２ 77. The method of any of claims 72 to 76, wherein the compound is present on the surface in a dosage of

78. 78. The method of claim 77, wherein the composition is applied to the infection and the wound closes within 12 weeks of the initial administration of the composition.

79. 79. The method of claim 78, wherein the BT composition is administered once a day or three times a week.

80. 80. The method of any of claims 72-79, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

81. 81. The method of claim 80, wherein the subject is administered multiple doses of the BT composition daily or weekly for a period of about 4 weeks.

82. The wound area is approximately 0.1 cm ２ ~Approx. 250cm ２ The method according to any one of claims 72 to 81, wherein

83. 1. A method for reducing wound size in a subject having a diabetic foot infection, comprising administering to the subject a composition comprising a therapeutically effective amount of BisEDT, wherein the composition is applied to the infection and the wound reduces in size from about 1% reduction compared to the original wound size to complete clearance of the wound within 12 weeks of initial administration of the composition.

84. 84. The method of claim 83, wherein the wound is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%.

85. 85. The method of claim 84, wherein the wound is reduced by about 50%.

86. 86. The method of any one of claims 83 to 85, wherein the composition is a suspension of microparticles comprising the BisEDT, and at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

87. 87. The method of any one of claims 83-86, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05-40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2-20 mM sodium phosphate.

88. The applied BT composition is about 1 μg / cm ２ ~Approx. 1,000,000μg / cm ２ The method of any one of claims 83 to 87, wherein the compound is present on the surface at a concentration of

89. The applied BT composition is about 50 μg / cm ２ ~Approx. 100μg / cm ２ The method of any of claims 83 to 88, wherein the compound is present on the surface at a concentration of

90. The applied BT composition is about 100 μg / cm ２ Concentrations greater than about 250 μg / cm ２ ~Approx. 5,000μg / cm ２ 90. The method of any of claims 83 to 89, wherein the compound is present on the surface in a dosage of

91. 91. The method of any one of claims 83 to 90, wherein the BT composition is administered three times a day, twice a day, once a day, every other day, once every three days, three times a week, once a week, once every two weeks, once a month, or once every two months.

92. 92. The method of claim 91, wherein the BT composition is administered once a day or three times a week.

93. 93. The method of any of claims 83-92, wherein the subject is administered multiple doses of the BT composition daily or weekly for a length of time ranging from about 1 week to about 12 weeks.

94. 94. The method of claim 93, wherein the subject is administered multiple doses of the BT composition daily or weekly for a period of about 4 weeks.

95. The wound area is approximately 0.1 cm ２ ~Approx. 250cm ２ The method according to any one of claims 83 to 94, wherein

96. 96. The method of any one of claims 83 to 95, wherein the wound surface area of ​​the wound is reduced by 50% 12 weeks after the first administration of the BT composition, and wherein the BT composition is BisEDT.

97. 97. The method of claim 96, wherein the wound surface area of ​​the wound is reduced by 50% four weeks after the initial administration of the BisEDT composition.

98. 98. The method of any one of claims 83 to 97, wherein the wound surface area is measured using digital photography or manual measurement.

99. 13. A method for preventing amputation and / or infection-related surgery in a subject with a diabetic foot infection, comprising administering to the subject a therapeutically effective amount of a BT composition.

100. 100. The method of claim 99, wherein the composition is a suspension of microparticles comprising the BisEDT, at least 70% of the microparticles having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

101. 101. The method of any one of claims 99-100, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05-40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2-20 mM sodium phosphate.

102. The applied BT composition is about 1 μg / cm ２ ~Approx. 1,000,000μg / cm ２ The method of any one of claims 99 to 101, wherein the compound is present on the surface at a concentration of

103. The applied BT composition is about 50 μg / cm ２ ~Approx. 100μg / cm ２ The method of any of claims 99 to 102, wherein the compound is present on the surface at a concentration of

104. The applied BT composition is about 100 μg / cm ２ Concentrations greater than about 250 μg / cm ２ ~Approx. 5,000μg / cm ２ The method of any one of claims 99 to 103, wherein the compound is present on the surface in a dosage of

105. 11. A method of treating a wound in a subject, the wound being 30 days or older, comprising administering to the subject a therapeutically effective amount of a BT composition.

106. The method of claim 105, wherein the subject has a diabetic foot infection.

107. 107. The method of claim 105 or 106, wherein the composition is a suspension of microparticles comprising BisEDT, and at least 70% of the microparticles have a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

108. 108. The method of any one of claims 105-107, wherein the BT composition further comprises about 0.05% to about 1.0% Tween 80® at about pH 7.4, about 0.05-40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2-20 mM sodium phosphate.

109. The applied BT composition is about 1 μg / cm ２ ~Approx. 1,000,000μg / cm ２ The method of any one of claims 105 to 108, wherein the compound is present on the surface at a concentration of

110. The applied BT composition is about 50 μg / cm ２ ~Approx. 100μg / cm ２ The method of any of claims 105 to 109, wherein the compound is present on the surface at a concentration of

111. The applied BT composition is about 100 μg / cm ２ Concentrations greater than about 250 μg / cm ２ ~Approx. 5,000μg / cm ２ The method of any of claims 105 to 110, wherein the compound is present on the surface in a dosage of

112. 112. The method of any of claims 105-111, wherein the wound is older than 2 months, older than 3 months, older than 4 months, older than 5 months, older than 6 months, older than 7 months, older than 8 months, older than 9 months, older than 10 months, older than 11 months, or older than 1 year.

113. 113. The method of claim 112, wherein the wound is more than two months old.

114. 113. The method of claim 112, wherein the wound is more than three months old.

115. 115. The method of any one of claims 1 to 114, wherein the BT composition comprises BisEDT, the composition being a suspension of microparticles comprising the BisEDT compound having a volume mean diameter (VMD) of about 0.4 μm to about 5 μm.

116. 116. The method of claim 115, wherein at least 70% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm.

117. 117. The method of claim 116, wherein the VMD is from about 0.4 μm to about 3 μm.

118. 117. The method of claim 116, wherein the VMD is from about 1.0 μm to about 2.0 μm.

119. 119. The method of any one of claims 116-118, wherein at least 70%, 75%, 80%, 90%, or 95% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm.

120. 120. The method of claim 119, wherein at least 80% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm.

121. 120. The method of claim 119, wherein at least 90% of the microparticles have a VMD of about 0.4 μm to about 3 μm, or about 0.5 μm to about 2 μm, or about 0.7 μm to about 2 μm, or about 0.8 μm to about 1.8 μm, or about 0.8 μm to about 1.6 μm, or about 0.9 μm to about 1.4 μm, or about 1.0 μm to about 2.0 μm, or about 1.0 μm to about 1.8 μm.

122. 120. The method of claim 119, wherein substantially all of the microparticles have a VMD of from about 0.4 μm to about 3 μm, or from about 0.5 μm to about 2 μm, or from about 0.7 μm to about 2 μm, or from about 0.8 μm to about 1.8 μm, or from about 0.8 μm to about 1.6 μm, or from about 0.9 μm to about 1.4 μm, or from about 1.0 μm to about 2.0 μm, or from about 1.0 μm to about 1.8 μm.

123. 123. The method of claims 119-122, wherein the VMD is from about 0.4 μm to about 3 μm.

124. The method of claims 119-122, wherein the VMD is from about 1.0 μm to about 2.0 μm.

125. 1. A pharmaceutical composition comprising a bismuth thiol (BT) composition comprising BisEDT suspended therein, said BT composition comprising a plurality of microparticles, said microparticles having a D90 of 1.9 μm or less.

126. A pharmaceutical composition comprising a bismuth thiol (BT) composition comprising BisEDT, the BT composition comprising a plurality of microparticles, the microparticles having a D90 of about 1.6 μm or less.

127. The pharmaceutical composition of claim 125, wherein at least 70% of the microparticles have a volume mean diameter of about 0.6 μm to about 2.5 μm.

128. The pharmaceutical composition of claim 125, wherein at least 90% of the microparticles have a volume mean diameter of about 0.6 μm to about 2.5 μm.

129. The pharmaceutical composition of claim 125, wherein the microparticles have a D50 of 1.0 μm or less.

130. The pharmaceutical composition of claim 125, wherein the microparticles have a D10 of 0.39 μm or less.

131. 132. The pharmaceutical composition of any one of claims 125-131, wherein the composition further comprises about 0.05% to about 1.0% Tween 80®, about 0.05-40 mM sodium chloride, optionally about 1% to about 10% methylcellulose, and optionally about 2-20 mM sodium phosphate at about pH 7.

4.

132. 132. A method for treating a topical wound, comprising administering to a subject in need thereof a therapeutically effective amount of the composition of any one of claims 125 to 131, wherein the composition is applied to an infection.

133. The infection is associated with a wound (e.g., an ulcer) and the subject experiences the following outcomes after completion of dosing: a) the wound is healed or substantially healed within 12 weeks (e.g., within 4 weeks) of the initial administration of the composition; and / or b) preventing amputations and / or infection-related surgery; and / or c) the wound partially or completely closes; and / or d) the wound is reduced in size from about 1% reduction compared to the original wound size to complete elimination of the wound; and / or e) the wound is 30 days or older and is healed or substantially healed.

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