Minocycline compounds for biodefense

Abstract

To provide methods of treating bacterial infections caused by biological agents including those which can be used as weapons.SOLUTION: A method comprises administering to a subject an effective amount of a compound, or a salt thereof, at a dose of about 10 mg to about 1,000 mg, where the compound is Compound A' having a structural formula in the figure, where the bacterial infection is caused by a bacterium which can be used as a biological weapon, thereby treating the infection in the subject.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] Related Applications This application claims priority to U.S. Provisional Application No. 62 / 137,719, filed May 24, 2015, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] 2. Background of the Invention

[0003] Biological agents, including various types of bacteria such as Bacillus anthracis and multidrug-resistant (MDR) Bacillus anthracis, Francisella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, and Rickettsia prowazekii, can be used as weapons and pose a material threat to the national security and public health of the United States. Tetracyclines have proven clinical utility as antibacterial agents. As a group, tetracyclines have well-established evidence of safety and efficacy. Tetracyclines exert their antibacterial action through multiple routes, including binding to the 30S subunit of the bacterial ribosome and inhibiting the binding of aminoacyl-tRNA. Tetracyclines are known to be effective against infections caused by a variety of pathogens. Tetracyclines are often applied in the treatment and prevention of diseases caused by these pathogens. Summary of the Invention

[0004] The most common tetracycline resistance mechanisms in Gram-positive and Gram-negative bacteria are ribosome protection and extrusion. Both of these mechanisms can be easily transferred between bacterial species, often associated with transferable genetic elements such as plasmids, transposons and integrons. Therefore, effective antimicrobial agents are needed for the prevention, prophylaxis and treatment of infections caused by biological agents, including those that can be used as weapons.

[0005] In some embodiments, the present invention provides a method of treating a bacterial infection in a subject in need thereof, comprising administering to the subject an effective amount of a compound or salt thereof at a dose of from about 10 mg to about 1000 mg, the compound being compound A′ having the following structural formula:

[0006] [ka] The bacterial infection is caused by a bacterium that could potentially be used as a biological weapon, and such infection is treated in a subject. In one embodiment, the compound is Compound A, which has the following structural formula:

[0007] [ka]

[0008] In some embodiments, the bacteria are resistant to antibiotics that are normally used to treat bacterial infections. In certain embodiments, the bacteria are in the form of a powder or aerosol. In other particular embodiments, the bacteria are capable of forming spores. In some embodiments, the bacteria can be spread as spores or via contamination of food or drinking water.

[0009] In some embodiments, the bacteria is selected from the group including: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0010] In another embodiment, the bacteria is selected from the group comprising: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0011] In one embodiment, the bacteria is selected from the group consisting of: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0012] In yet another embodiment, the bacteria is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0013] In yet another embodiment, the bacterium is selected from the group consisting of Yersinia pestis, Burkholderia mallei, and B. anthracis.

[0014] In certain embodiments, the compound is administered once per day or twice per day.

[0015] In some embodiments, the compound is administered intravenously or orally.

[0016] In one embodiment, the compound is administered intravenously at a dose of about 50 mg to about 200 mg. In yet another embodiment, the compound is administered at a dose of about 100 mg.

[0017] In another embodiment, the compound is administered orally in a dose of about 100 mg to about 300 mg.

[0018] In certain aspects, the methods of the invention include administering a compound, e.g., Compound A' or Compound A, for at least 3 days, at least 7 days, at least 14 days, at least 21 days, at least 30 days, or at least 60 days. In certain embodiments, the methods include administering the compound for about 30 days or about 60 days.

[0019] In some embodiments, the method includes administering to the subject one or more loading doses of a compound, followed by one or more maintenance doses of the compound, hi certain embodiments, the one or more loading doses can be greater than the one or more maintenance doses.

[0020] In certain embodiments, the loading dose is an intravenous dose and the maintenance dose is an oral dose. In other particular embodiments, the loading dose is an intravenous dose and the maintenance dose is an intravenous dose. In yet other particular embodiments, the loading dose is an oral dose and the maintenance dose is an oral dose.

[0021] In certain embodiments, the subject is a human.

[0022] In some embodiments, the present invention also provides a method of preventing a bacterial infection in a subject in need thereof, comprising administering to the subject an effective amount of a compound or a salt thereof at a dose of from about 10 mg to about 1000 mg, the compound being Compound A' having the following structural formula:

[0023] [ka] The bacterial infection is caused by a bacterium that could be used as a biological weapon, and such infection is to be prevented in the subject.

[0024] In one embodiment, the compound is Compound A, which has the following structural formula:

[0025] [ka]

[0026] In some embodiments, the bacteria is resistant to antibiotics that are typically used to treat bacterial infections. In certain embodiments, the bacteria is in the form of a powder or aerosol. In other particular embodiments, the bacteria is capable of forming spores.

[0027] In some embodiments, the bacteria is selected from the group including: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae), Salmonella (S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0028] In another embodiment, the bacteria is selected from the group comprising: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0029] In one embodiment, the bacteria is selected from the group consisting of: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0030] In yet another embodiment, the bacteria is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0031] In yet another embodiment, the bacterium is selected from the group consisting of Yersinia pestis, Burkholderia mallei, and B. anthracis.

[0032] In some embodiments, the dose is from about 50 mg to about 200 mg, for example, from about 75 mg to about 110 mg. In some embodiments, the dose is about 100 mg.

[0033] In certain embodiments, the compound is administered once per day or twice per day.

[0034] In certain embodiments, the compound is administered intravenously or orally.

[0035] In certain aspects, the methods of the invention include administering a compound, e.g., Compound A' or Compound A, for at least 3 days, at least 7 days, at least 14 days, at least 21 days, at least 30 days, or at least 60 days. In certain embodiments, the methods include administering the compound for about 30 days or about 60 days.

[0036] In some embodiments, the method includes administering to the subject one or more loading doses of a compound, followed by one or more maintenance doses of the compound, hi certain embodiments, the one or more loading doses can be greater than the one or more maintenance doses.

[0037] In certain embodiments, the loading dose is an intravenous dose and the maintenance dose is an oral dose. In other particular embodiments, the loading dose is an intravenous dose and the maintenance dose is an intravenous dose. In yet other particular embodiments, the loading dose is an oral dose and the maintenance dose is an oral dose.

[0038] In certain embodiments, the subject is a human. [Brief description of the drawings]

[0039] [Figure 1]FIG. 1 is a graph showing percent survival in a lethal Y. pestis post-exposure prophylaxis (PEP) mouse model following treatment with Compound A (omadacycline), doxycycline and ciprofloxacin. [Diagram 2] FIG. 2 is a graph depicting percent survival in a B. anthracis post-exposure prophylaxis (PEP) mouse model following treatment with Compound A (omadacycline), doxycycline and ciprofloxacin. [Diagram 3] FIG. 3 is a graph depicting percent survival in a lethal B. mallei post-exposure prophylaxis (PEP) mouse model following treatment with Compound A (omadacycline), doxycycline and moxifloxacin. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0040] Detailed Description of the Invention The present invention relates to the discovery that 9-[(2,2-dimethyl-propylamino)-methyl]minocycline (omadacycline, OMC) is effective for treating or preventing infections caused by a variety of bacterial species that have the potential to be used as biological weapons.

[0041] In one embodiment, the invention pertains, at least in part, to a method of treating a bacterial infection in a subject, the method comprising administering to the subject an effective amount of Compound A', or a salt thereof:

[0042] [ka]

[0043] In one embodiment, the invention pertains, at least in part, to a method of treating a bacterial infection in a subject, the method comprising administering to the subject a compound or a salt thereof at a dose of from about 10 mg to about 1000 mg, the compound being Compound A' having the following structural formula:

[0044] [ka]

[0045] In one embodiment, the invention relates to a method of preventing, at least in part, a bacterial infection in a subject, the method comprising administering to the subject an effective amount of Compound A' or a salt thereof:

[0046] [ka]

[0047] In one embodiment, the invention relates to a method of preventing, at least in part, a bacterial infection in a subject, the method comprising administering to the subject a compound or a salt thereof at a dose of from about 10 mg to about 1000 mg, the compound being Compound A' having the following structural formula:

[0048] [ka]

[0049] In one embodiment, the invention pertains, at least in part, to a method of treating a bacterial infection in a subject, the method comprising administering to the subject an effective amount of Compound A, or a salt thereof:

[0050] [ka]

[0051] In one embodiment, the invention pertains, at least in part, to a method of treating a bacterial infection in a subject, the method comprising administering to the subject a compound or a salt thereof at a dose of from about 10 mg to about 1000 mg, the compound being Compound A having the following structural formula:

[0052] [ka]

[0053] In one embodiment, the invention relates to a method of preventing, at least in part, a bacterial infection in a subject, the method comprising administering to the subject an effective amount of Compound A or a salt thereof:

[0054] [ka]

[0055] In one embodiment, the invention relates to a method of preventing, at least in part, a bacterial infection in a subject, the method comprising administering to the subject a compound or a salt thereof at a dose of from about 10 mg to about 1000 mg, the compound being Compound A having the following structural formula:

[0056] [ka]

[0057] In certain embodiments, the invention pertains, at least in part, to a method of treating an infection in a subject or a method of preventing an infection in a subject, the method comprising administering to the subject an effective amount of Compound A' or Compound A, the infection being caused by a bacteria that may be used as a biological weapon.

[0058] In certain embodiments, bacteria that may be used as biological weapons are bacteria that have one or more characteristics, including, but not limited to, being easy to manufacture and disseminate, being easily transmissible from person to person, having a moderate or high potential for morbidity, having a moderate or high potential for mortality, having the potential to cause public panic and social unrest, requiring special actions in public health preparedness, and requiring specially enhanced diagnostics and disease surveillance.

[0059] In some embodiments, bacteria that may be used as a biological weapon are stable or viable (e.g., capable of performing all or some of the normal biological functions of growth, sporulation, and infection of a subject) under a variety of conditions, including harsh conditions (e.g., high temperature, low temperature, high pressure, low pressure, acidic or alkaline conditions, wetness, dryness, and radiation). In some embodiments, bacteria that may be used as a biological weapon are capable of infecting a subject under a variety of conditions. In some embodiments, bacteria that may be used as a biological weapon are stable or viable under temperature conditions greater than 25°C, 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C, 100°C, 125°C, 150°C, 175°C, or 200°C. In some embodiments, the bacteria that may be used as a biological weapon are stable or viable under temperature conditions below 25° C., 20° C., 10° C., 5° C., 0° C., −10° C., −20° C., −30° C., −40° C., −50° C., −60° C., −70° C., −100° C., or −150° C. In some embodiments, the bacteria that may be used as a biological weapon are stable or viable under temperature conditions below 5 x 10 5 Pa, 10 x 10 5 Pa, 15 x 10 5 Pa, 20 x 10 5 Pa, 30 x 10 5 Pa, 40 x 10 5 Pa, 50 x 10 5 Pa, 75 x 10 5 Pa, or 100 x 10 5In one embodiment, the bacteria that may be used as a biological weapon are stable or viable under pressure conditions greater than 0.5 x 10 5 Pa, 0.2 x 10 5 Pa, 0.1 x 10 5 Pa, 0.05 x 10 5 Pa, 0.02 x 10 5 Pa, 0.01 x 10 5 Pa, 0.005 x 10 5 Pa, 0.002 x 10 5 Pa, or 0.001 x 10 5 In some embodiments, the bacteria that may be used as a biological weapon are stable or viable under pressure conditions below 10 Pa. In some embodiments, the bacteria that may be used as a biological weapon are stable or viable under pH conditions above 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, or 14.0. In some embodiments, the bacteria that may be used as a biological weapon are stable or viable under pH conditions below 6.0, 5.5, 5.0, 4.5, 4.0, 3.5, 3.0, 2.5, 2.0, 1.5, 1.0, 0.5, or 0.0. In some embodiments, the bacteria that may be used as a biological weapon are stable or viable under relative humidity conditions of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%. In one embodiment, the bacteria that may be used as a biological weapon are stable or viable under deep UV, x-ray, alpha, beta, or gamma irradiation, hi another embodiment, the bacteria are capable of infecting a subject after being treated with any combination of the above conditions.

[0060] In certain embodiments, bacteria that may be used as biological weapons are capable of forming spores.

[0061] In one embodiment, bacteria that may be used as a biological weapon may be spread as spores. In another embodiment, bacteria that may be used as a biological weapon may be spread through contamination of food or drinking water. In yet other embodiments, bacteria that may be used as a biological weapon may be spread by insects (e.g., fleas, lice, and ticks) and / or rodents (mice or rats).

[0062] In some embodiments, the bacteria, which may be used as a biological weapon, may be dispersed into the air or liquid. In some embodiments, the bacteria is in the form of an aerosol (e.g., the bacteria is formulated as an aerosol). In other embodiments, the bacteria is in powder form (e.g., the bacteria is formulated as a powder).

[0063] In some embodiments, bacteria that may be used as biological weapons include bacteria that are resistant to existing antibiotics, i.e., antibiotics that are commonly used to treat bacterial infections. In some embodiments, such antibiotics include, for example, tetracycline antibiotics and other antibiotics, including tetracycline, doxycycline, minocycline, sancycline, methacycline, chlortetracycline, and deoxytetracycline, and combinations thereof, and including, but not limited to, methicillin, oxacillin, vancomycin, penicillin, linezolid, ciprofloxacin, ceftazidime, and azithromycin. In yet other embodiments, bacteria that may be used as biological weapons include bacteria that are resistant to tetracycline, minocycline, and / or doxycycline.

[0064] In certain embodiments, bacteria that may be used as biological weapons include, but are not limited to: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0065] In another embodiment, bacteria that may be used as biological weapons include, but are not limited to: Bacteria belonging to the species Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0066] In certain embodiments, bacteria that may be used as biological weapons include, but are not limited to: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), vulnificus), bacteria belonging to the genus Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0067] In another embodiment, bacteria that may be used as biological weapons include, but are not limited to: Bacteria belonging to the species Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), vulnificus), bacteria belonging to the genus Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0068] In yet other embodiments, bacteria that may be used as biological weapons include, but are not limited to: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0069] In yet another embodiment, bacteria that may be used as biological weapons include, but are not limited to: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, and Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0070] In other embodiments, bacteria that may be used as a biological weapon include, but are not limited to, bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, and Rickettsia prowazekii. In yet other embodiments, bacteria that may be used as a biological weapon include, but are not limited to, bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, and Rickettsia prowazekii. In yet another embodiment, bacteria that may be used as a biological weapon are bacteria belonging to the species Bacillus anthracis, or multi-drug resistant (MDR) strains of Bacillus anthracis.

[0071] The genus Bacillus includes Bacillus anthracis (the causative agent of anthrax), Bacillus cereus, Bacillus weihenstephanensis (a foodborne pathogen), Bacillus thuringiensis (an insect pathogen), and Bacillus mycoides.

[0072] In certain embodiments, bacteria that may be used as biological weapons include bacteria from the Bacillus cereus group (e.g., Bacillus anthracis and multidrug resistant (MDR) anthrax), Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Brucella species, Shigella species, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia typhi, and the like. prowazekii, diarrheagenic E. coli, pathogenic Vibrios, Salmonella, Campylobacter jejuni, Yersinia enterocolitica, and Listeria monocytogenes.In certain embodiments, bacteria that may be used as biological weapons include bacteria of the Bacillus cereus group (e.g., Bacillus anthracis and multidrug resistant (MDR) anthrax), Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Brucella species, Shigella species, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, diarrheagenic E. coli, pathogenic Vibrio species, and the like. Examples of suitable pathogens include, but are not limited to, Vibrios, Salmonella, Campylobacter jejuni, Yersinia enterocolitica, and Listeria monocytogenes.

[0073] In certain embodiments, bacteria that may be used as biological weapons include Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Brucella species, Shigella species, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, diarrheagenic E. coli, pathogenic Vibrios, Salmonella, Campylobacter jejuni, and the like. Examples of suitable pathogenic bacteria include, but are not limited to, Yersinia jejuni, Yersinia enterocolitica, and Listeria monocytogenes.In certain embodiments, bacteria that may be used as biological weapons include Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Brucella species, Shigella species, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, diarrheagenic E. coli, pathogenic Vibrios, Salmonella, Campylobacter jejuni, Yersinia enterocolitica, and the like. enterocolitica, and Listeria monocytogenes.

[0074] In yet other embodiments, bacteria that may be used as biological weapons include, but are not limited to, bacteria of the Bacillus cereus group (e.g., Bacillus anthracis and multidrug resistant (MDR) anthrax), Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, and Rickettsia prowazekii. In yet another embodiment, bacteria that may be used as biological weapons include, but are not limited to, bacteria of the Bacillus cereus group (e.g., Bacillus anthracis and multidrug resistant (MDR) anthrax), Franciscella tularensis, Yersinia pestis, Burkholderia mallei, and Rickettsia prowazekii.

[0075] In yet another embodiment, bacteria that may be used as a biological weapon include, but are not limited to, Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, and Rickettsia prowazekii. In yet another embodiment, bacteria that may be used as a biological weapon include, but are not limited to, Franciscella tularensis, Yersinia pestis, Burkholderia mallei, and Rickettsia prowazekii.

[0076] In other embodiments, the bacterium that may be used as a biological weapon is Bacillus anthracis or multi-drug resistant (MDR) anthrax.

[0077] The Bacillus cereus group of bacteria consists of Bacillus anthracis (the causative agent of anthrax), Bacillus cereus, and Bacillus weihenstephanensis (foodborne pathogens), Bacillus thuringiensis (an insect pathogen), and Bacillus mycoides.

[0078] In certain embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. anthracis, Y. pestis, F. tularensis, B. mallei, or B. pseudomallei. In certain embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. anthracis. In other embodiments, the bacteria that may be used as a biological weapon do not belong to the species Y. pestis. In yet other specific embodiments, the bacteria that may be used as a biological weapon do not belong to the species F. tularensis. In yet other specific embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. mallei. In yet other specific embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. pseudomallei.

[0079] In certain embodiments, the bacteria that may be used as a biological weapon are not bacteria that may be causative agents of food-borne diseases. In certain embodiments, the bacteria that may be used as a biological weapon do not belong to diarrheagenic E. coli strains. In another specific embodiment, the bacteria that may be used as a biological weapon do not belong to the genus Salmonella (e.g., S. bongori and S. enterica). In yet another specific embodiment, the bacteria that may be used as a biological weapon do not belong to the species Campylobacter jejuni.

[0080] In another embodiment, the bacteria that may be used as a biological weapon are bacteria that may be causative agents of food-borne diseases. In a particular embodiment, the bacteria that may be used as a biological weapon belong to a diarrheagenic E. coli strain. In another particular embodiment, the bacteria that may be used as a biological weapon belong to the genus Salmonella (e.g., S. bongori and S. enterica). In yet another particular embodiment, the bacteria that may be used as a biological weapon belong to the species Campylobacter jejuni.

[0081] A "foodborne illness" or "food-borne disease" or "food poisoning" is any illness resulting from the ingestion of food contaminated with, for example, bacteria. In certain embodiments, the contaminating bacteria can cause infection and irritation of the digestive tract. In some embodiments, the contaminating bacteria can belong to diarrheagenic E. coli strains; Campylobacter jejuni species; or the genus Salmonella (e.g., S. bongori and S. enterica).

[0082] In certain embodiments, bacteria that may be used as biological weapons include, but are not limited to, gram-positive pathogens, gram-negative pathogens, anaerobic pathogens, or atypical pathogens, or combinations thereof. In other embodiments, bacteria that may be used as biological weapons include Methicillin-susceptible Staphylococcus aureus (MSSA), Methicillin-resistant Staphylococcus aureus (MRSA), Oxacillin-susceptible Staphylococcus aureus, Oxacillin-resistant Staphylococcus aureus, Oxacillin-resistant Coagulase-negative Staphylococcus, Enterococcus faecalis, Enterococcus faecium, Vancomycin-susceptible Enterococcus faecium, Vancomycin-resistant Enterococcus faecium, Vancomycin-susceptible Enterococcus faecalis, Vancomycin-resistant Enterococcus faecium, Streptococcus pneumoniae, Penicillin-susceptible Streptococcus pneumoniae, Penicillin-resistant Streptococcus pneumoniae (PRSP), Streptococcus pyogenes ( ... pyogenes, Streptococcus agalactiae, Haemophilus influenzae, Moraxella catarrhalis, Neisseria gonorrhoeae, Escherichia coli, Shigella spp., Salmonella spp., Klebsiella pneumoniae, Enterobacter aerogenes, Enterobacter cloacae, Serratia marcescens, Acinetobacter baumannii, Stenotrophomonas maltophilia, Bacteroides fragilis fragilis, Clostridium perfringensExamples of bacteria that may be present include, but are not limited to, bacteria belonging to the species Chlamydia pneumoniae, Legionella pneumophila, Proteus mirabilis, Pseudomonas aeruginosa, and Burkholderia cepacia.

[0083] In one embodiment, the invention pertains, at least in part, to a method of treating a bacterial infection in a subject, the method comprising administering to the subject an effective amount of Compound A', or a salt thereof:

[0084] [ka] The bacterial infection is caused by a bacterium selected from the group consisting of: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0085] In another embodiment, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0086] In certain embodiments, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), vulnificus), bacteria belonging to the genus Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0087] In certain embodiments, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), vulnificus), bacteria belonging to the genus Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0088] In yet another embodiment, the bacteria that may be used as a biological weapon are selected from the group consisting of: bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacillus (eg, B. anthracis, including multidrug resistant (MDR) anthrax strains).

[0089] In yet another embodiment, the bacteria that may be used as a biological weapon are selected from the group consisting of: bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Rickettsia prowazekii; and Bacillus (eg, B. anthracis, including multidrug resistant (MDR) anthrax strains).

[0090] In another embodiment, the bacteria that may be used as a biological weapon are selected from the group consisting of bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, and Rickettsia prowazekii. In yet another embodiment, the bacteria that may be used as a biological weapon are selected from the group consisting of bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, and Rickettsia prowazekii. In another embodiment, the bacteria that may be used as a biological weapon are bacteria belonging to the species B. anthracis, or a multi-drug resistant (MDR) strain of B. anthracis.

[0091] In certain embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. anthracis, Y. pestis, F. tularensis, B. mallei, or B. pseudomallei. In certain embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. anthracis. In other embodiments, the bacteria that may be used as a biological weapon do not belong to the species Y. pestis. In other specific embodiments, the bacteria that may be used as a biological weapon do not belong to the species F. tularensis. In other specific embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. mallei. In yet other specific embodiments, the bacteria that may be used as a biological weapon do not belong to B. pseudomallei.

[0092] In certain embodiments, the bacteria that may be used as a biological weapon are not bacteria that may be causative pathogens of foodborne diseases. In certain embodiments, the bacteria that may be used as a biological weapon do not belong to diarrheagenic E. coli strains. In another specific embodiment, the bacteria that may be used as a biological weapon do not belong to the genus Salmonella (e.g., S. bongori and S. enterica). In yet another specific embodiment, the bacteria that may be used as a biological weapon do not belong to the species Campylobacter jejuni.

[0093] It will be appreciated that for all of the embodiments described, the compounds of the invention, for example Compound A', can be administered in doses from about 10 mg to about 1000 mg.

[0094] In one embodiment, the invention relates to a method of preventing, at least in part, a bacterial infection in a subject, the method comprising administering to the subject an effective amount of Compound A', or a salt thereof, wherein:

[0095] [ka] The bacterial infection is caused by a bacterium selected from the group consisting of: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0096] In another embodiment, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0097] In certain embodiments, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), vulnificus), bacteria belonging to the genus Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0098] In certain embodiments, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), vulnificus), bacteria belonging to the genus Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0099] In yet another embodiment, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0100] In yet other embodiments, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, and Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0101] In another embodiment, the bacteria that may be used as a biological weapon are selected from the group consisting of bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, and Rickettsia prowazekii. In yet another embodiment, the bacteria that may be used as a biological weapon are selected from the group consisting of bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, and Rickettsia prowazekii. In another embodiment, the bacteria that may be used as a biological weapon are bacteria belonging to the species B. anthracis, or a multi-drug resistant (MDR) strain of B. anthracis.

[0102] In certain embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. anthracis, Y. pestis, F. tularensis, B. mallei, or B. pseudomallei. In certain embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. anthracis. In other embodiments, the bacteria that may be used as a biological weapon do not belong to the species Y. pestis. In other specific embodiments, the bacteria that may be used as a biological weapon do not belong to the species F. tularensis. In other specific embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. mallei. In yet other specific embodiments, the bacteria that may be used as a biological weapon do not belong to B. pseudomallei.

[0103] In certain embodiments, the bacteria that may be used as a biological weapon are not bacteria that may be causative pathogens of foodborne diseases. In certain embodiments, the bacteria that may be used as a biological weapon do not belong to diarrheagenic E. coli strains. In another specific embodiment, the bacteria that may be used as a biological weapon do not belong to the genus Salmonella (e.g., S. bongori and S. enterica). In yet another specific embodiment, the bacteria that may be used as a biological weapon do not belong to the species Campylobacter jejuni.

[0104] It will be appreciated that for all of the embodiments described, the compounds of the invention, for example Compound A', can be administered in doses from about 10 mg to about 1000 mg.

[0105] In one embodiment, the invention relates, at least in part, to a method of treating a bacterial infection in a subject, the method comprising administering to the subject an effective amount of Compound A, or a salt thereof: [ka] The bacterial infection is caused by a bacterium selected from the group consisting of: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0106] In another embodiment, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0107] In certain embodiments, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), vulnificus), bacteria belonging to the genus Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0108] In certain embodiments, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), vulnificus), bacteria belonging to the genus Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0109] In yet another embodiment, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0110] In yet other embodiments, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, and Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0111] In another embodiment, the bacteria that may be used as a biological weapon are selected from the group consisting of bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, and Rickettsia prowazekii. In yet another embodiment, the bacteria that may be used as a biological weapon are selected from the group consisting of bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, and Rickettsia prowazekii. In another embodiment, the bacteria that may be used as a biological weapon are bacteria belonging to the species B. anthracis, or a multi-drug resistant (MDR) strain of B. anthracis.

[0112] In certain embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. anthracis, Y. pestis, F. tularensis, B. mallei, or B. pseudomallei. In certain embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. anthracis. In other embodiments, the bacteria that may be used as a biological weapon do not belong to the species Y. pestis. In other specific embodiments, the bacteria that may be used as a biological weapon do not belong to the species F. tularensis. In other specific embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. mallei. In yet other specific embodiments, the bacteria that may be used as a biological weapon do not belong to B. pseudomallei.

[0113] In certain embodiments, the bacteria that may be used as a biological weapon are not bacteria that may be causative pathogens of foodborne diseases. In certain embodiments, the bacteria that may be used as a biological weapon do not belong to diarrheagenic E. coli strains. In another specific embodiment, the bacteria that may be used as a biological weapon do not belong to the genus Salmonella (e.g., S. bongori and S. enterica). In yet another specific embodiment, the bacteria that may be used as a biological weapon do not belong to the species Campylobacter jejuni.

[0114] It will be appreciated that for all of the embodiments described, the compounds of the invention, for example Compound A, can be administered in dosages from about 10 mg to about 1000 mg.

[0115] In one embodiment, the invention relates to a method of preventing, at least in part, a bacterial infection in a subject, the method comprising administering to the subject an effective amount of Compound A or a salt thereof:

[0116] [ka] The bacterial infection is caused by a bacterium selected from the group consisting of: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0117] In another embodiment, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0118] In certain embodiments, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), vulnificus), bacteria belonging to the genus Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0119] In certain embodiments, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. parahaemolyticus, and V. vulnificus), vulnificus), bacteria belonging to the genus Salmonella (e.g., S. bongori and S. enterica); and A bacterium belonging to the diarrheagenic Escherichia coli (E. coli) strains.

[0120] In yet another embodiment, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0121] In yet other embodiments, the bacteria that may be used as a biological weapon is selected from the group consisting of: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, and Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (eg, B. anthracis, including multidrug-resistant (MDR) anthrax strains).

[0122] In another embodiment, the bacteria that may be used as a biological weapon are selected from the group consisting of bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, and Rickettsia prowazekii. In yet another embodiment, the bacteria that may be used as a biological weapon are selected from the group consisting of bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, and Rickettsia prowazekii. In another embodiment, the bacteria that may be used as a biological weapon are bacteria belonging to the species B. anthracis, or a multi-drug resistant (MDR) strain of B. anthracis.

[0123] In certain embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. anthracis, Y. pestis, F. tularensis, B. mallei, or B. pseudomallei. In certain embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. anthracis. In other embodiments, the bacteria that may be used as a biological weapon do not belong to the species Y. pestis. In other specific embodiments, the bacteria that may be used as a biological weapon do not belong to the species F. tularensis. In other specific embodiments, the bacteria that may be used as a biological weapon do not belong to the species B. mallei. In yet other specific embodiments, the bacteria that may be used as a biological weapon do not belong to B. pseudomallei.

[0124] In certain embodiments, the bacteria that may be used as a biological weapon are not bacteria that may be causative pathogens of foodborne diseases. In certain embodiments, the bacteria that may be used as a biological weapon do not belong to diarrheagenic E. coli strains. In another specific embodiment, the bacteria that may be used as a biological weapon do not belong to the genus Salmonella (e.g., S. bongori and S. enterica). In yet another specific embodiment, the bacteria that may be used as a biological weapon do not belong to the species Campylobacter jejuni.

[0125] It will be appreciated that for all of the embodiments described, the compounds of the invention, for example Compound A, can be administered in dosages from about 10 mg to about 1000 mg.

[0126] In some embodiments, treating bacterial infection in a subject comprises administering a compound of the present invention after the subject is exposed to bacteria, but before the subject develops symptoms of bacterial infection.In some embodiments, the compound of the present invention is administered 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, or 2 weeks after the subject is exposed to bacteria, but before the subject develops symptoms of bacterial infection.

[0127] In another embodiment, treating a bacterial infection in a subject comprises administering a compound of the present invention after the subject develops symptoms following exposure to the bacteria. In some embodiments, the compound of the present invention is administered 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, or 2 weeks after the subject develops symptoms of a bacterial infection.

[0128] In another embodiment, treating a bacterial infection in a subject comprises administering a compound of the present invention after the subject is suspected to have been exposed to bacteria, but before the subject develops any symptoms of bacterial infection. In some embodiments, the compound of the present invention is administered 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, or 2 weeks after the subject is suspected to have been exposed to bacteria, but before the subject develops any symptoms.

[0129] "Suspected exposure" means that there is a reasonable, if not obvious, likelihood (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%) that the subject has been exposed to the bacteria and is therefore at risk for bacterial infection. In some embodiments, "suspected exposure" means that there is a greater than 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% likelihood that the subject has been exposed to the bacteria and is therefore at risk for bacterial infection. For example, "suspected exposure" means that there is a greater than 50% likelihood that the subject has been exposed to the microorganism.

[0130] A "symptom" of bacterial infection can be any indication that a subject exposed or suspected to be exposed to the bacteria is not normal, healthy, or comfortable, regardless of the subject's subjective perception or sensation. "Symptoms" include, but are not limited to, headache, stomach pain, abdominal cramps, abdominal pain, muscle pain, fever, diarrhea, vomiting, cough, weakness, fatigue, pain, skin rash or swelling, sores on any part of the body (e.g., skin, head, eyes, ears, mouth, torso, limbs, arms, hands, legs, feet, etc.), and abnormalities of any tissue or organ (e.g., skin, bones, blood, lymph, intestines, stomach, pancreas, brain, heart, lungs, liver, spleen, kidneys, bladder, ovaries, etc.).

[0131] In some embodiments, the prevention of bacterial infection in a subject comprises administering the compound of the present invention before the subject is exposed to bacteria.In some embodiments, the compound of the present invention is administered 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, or 2 weeks before the subject is exposed.In another embodiment, the prevention of bacterial infection in a subject comprises administering the compound of the present invention before or after an event that increases the risk of the subject being exposed to bacteria.These events include, but are not limited to, terrorist attacks with biological weapons, and the subject entering a dangerous area such as a battlefield. In some embodiments, the compounds of the present invention are administered to the subject 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, or 2 weeks prior to the event. In some embodiments, the compounds of the present invention are administered to the subject 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, or 2 weeks after the event.

[0132] In certain embodiments, the methods of the invention can further include identifying a subject at risk of exposure to a bacteria that could be used as a biological weapon prior to administering a compound of the invention. Subjects at risk of exposure to bacteria that could be used as biological weapons include, but are not limited to, subjects traveling to, entering, or present in conflict zones (e.g., battlefields and operational zones), such as military personnel, intelligence personnel, and military animals; subjects engaged or about to engage in law enforcement operations, such as government officials (e.g., police officers, government investigators, and Secret Service members) and other personnel (e.g., doctors, nurses, and rescue workers), and animals used in such operations; and subjects in geographic areas prone to being targeted by terrorist attacks (e.g., metropolitan areas, cities, populated areas (e.g., greater than 100,000 people, greater than 200,000 people, greater than 500,000 people, greater than 1 million people, greater than 2 million people, greater than 5 million people, and greater than 10 million people)), and locations or areas where damage thereto could pose a threat to national security or public health (e.g., nuclear plants, chemical plants, airports, and hospitals).

[0133] "Expose," "exposure," or "exposed" means that a subject comes into some contact with a bacterium or a component thereof (e.g., bacterial cell wall, bacterial cell membrane, bacterial nucleic acid, bacterial polynucleotide, bacterial protein, bacterial polypeptide, bacterial spore, and bacterial toxin). For example, a subject can be exposed to a bacterium or a component thereof by ingesting, inhaling, or touching anything that contains the bacterium or a component thereof. For example, a bacterial component has the ability to cause infection or symptoms of infection in a subject. A bacterial component is, for example, a bacterial spore.

[0134] In one embodiment, the present invention relates to a method of treating a bacterial infection in a subject, the subject being exposed to or suspected of being exposed to bacteria or a component thereof, comprising administering to the subject an effective amount of Compound A' or Compound A, or a salt thereof. In another embodiment, the present invention also relates to a method of treating a bacterial infection in a subject, the subject being exposed to or suspected of being exposed to bacteria or a component thereof, comprising administering to the subject Compound A' or Compound A, or a salt thereof, in a dose of about 10 mg to about 1000 mg. In one embodiment, the present invention also relates to a method of preventing a bacterial infection in a subject, the subject being at risk of being exposed to bacteria or a component thereof, comprising administering to the subject an effective amount of Compound A' or Compound A, or a salt thereof. In another embodiment, the present invention also relates to a method for preventing bacterial infection in a subject, the subject being at risk of exposure to bacteria or a component thereof, the method comprising administering to the subject an effective amount of Compound A' or Compound A, or a salt thereof, at a dose of about 10 mg to about 1000 mg. In one embodiment, the bacteria or a component thereof is formulated as an aerosol or powder. In one embodiment, the bacterial component is a bacterial spore.

[0135] A compound of the invention, e.g., Compound A, can be administered to a subject by any method of administration that can achieve a level of Compound A in the subject that is effective to treat or prevent an infection. In one embodiment, a compound of the invention is administered orally. In another embodiment, a compound of the invention is administered intravenously. In another embodiment, a compound of the invention is administered intraperitoneally. In yet another embodiment, a compound of the invention is administered subcutaneously.

[0136] In some embodiments, the compounds of the invention are administered at 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 12 mg / kg, 14 mg / kg, 16 mg / kg, 18 mg / kg, 20 mg / kg, 22 mg / kg, 24 mg / kg, 26 mg / kg, 28 mg / kg, 30 mg / kg, 32 mg / kg, 34 mg / kg, 36 mg / kg, 38 mg / kg, 40 mg / kg, 42 mg / kg, 44 mg / kg, 46 mg / kg, 48 mg / kg, 50 mg / kg, 52 mg / kg, 54 mg / kg, 56 mg / kg, 58 mg / kg, 60 mg / kg, 62 mg / kg, 64 mg / kg, 66 mg / kg, 68 mg / kg, 70 mg / kg, 72 mg / kg, 74 mg / kg, 76 mg / kg, 78 mg / kg, 79 mg / kg, 80 mg / kg, 81 mg / kg, 82 mg / kg, 83 mg / kg, 84 mg / kg, 85 mg / kg, 86 mg / kg, 87 mg / kg, 88 mg / kg, 89 mg / kg, 90 mg / kg, 91 mg / kg, 92 mg / kg, 93 mg / kg, 94 mg / kg, 95 mg / kg, 96 mg / kg, 97 mg / kg, 98 mg / kg, 99 mg / kg, 100 mg / kg, 102 mg / kg, 104 mg / kg, 106 mg / kg, 108 mg / kg, 109 mg / kg, 110 mg In some embodiments, the therapeutic agent may be administered at a dose of 76 mg / kg, 78 mg / kg, 80 mg / kg, 82 mg / kg, 84 mg / kg, 86 mg / kg, 88 mg / kg, 90 mg / kg, 92 mg / kg, 94 mg / kg, 96 mg / kg, 98 mg / kg, 100 mg / kg, 110 mg / kg, 120 mg / kg, 130 mg / kg, 140 mg / kg, 150 mg / kg, 160 mg / kg, 170 mg / kg, 180 mg / kg, 190 mg / kg, or 200 mg / kg.

[0137] It is to be understood that dosage ranges including the above dosages are also included in the present invention. For example, any of the above dosages may be in the lower or upper portion of a dosage range included in the present invention. It is further to be understood that any list or collection of numerical values ​​used throughout this specification is intended to encompass numerical ranges in which any of the recited numerical values ​​may be in the lower or upper portion of a range. These ranges are intended to be included in the present invention.

[0138] In some embodiments, a compound of the invention, e.g., Compound A' or Compound A, can be administered in a dose of about 10 to about 1000 mg, about 20 to about 750 mg, about 50 to about 500 mg, about 75 to about 400 mg, about 100 to about 300 mg, about 110 to about 290 mg, about 120 to about 280 mg, about 130 to about 270 mg, about 140 to about 260 mg, about 150 to about 250 mg, about 160 to about 240 mg, about 170 mg to about 230 mg, about 180 mg to about 220 mg, about 190 mg to about 210 mg, or about 200 mg. In another embodiment, a compound of the invention, e.g., Compound A' or Compound A, can be administered intravenously at a dose of about 5 to about 500 mg, about 10 to about 400 mg, about 25 to about 300 mg, about 50 to about 200 mg, about 50 to about 150 mg, about 60 to about 140 mg, about 70 to about 130 mg, about 80 mg to about 120 mg, about 90 mg to about 110 mg, or about 100 mg. In certain embodiments, a compound of the invention, e.g., Compound A' or Compound A, can be orally administered in a dose of about 5 to about 800 mg, about 10 to about 700 mg, about 25 to 600 mg, about 50 to about 500 mg, about 100 to about 400 mg, about 150 to about 350 mg, about 200 to about 340 mg, about 250 to about 330 mg, about 270 to about 320 mg, about 280 to about 310 mg, or about 300 mg.

[0139] In one embodiment, the compound of the present invention, e.g., Compound A' or Compound A, can be administered intravenously at a dose of about 100 mg, about 200 mg, or about 300 mg. In another embodiment, the compound of the present invention, e.g., Compound A' or Compound A, can be administered orally at a dose of about 300 mg, about 600 mg, or about 900 mg.

[0140] In one embodiment, the oral dose of a compound of the invention, eg, Compound A' or Compound A, is 3-fold greater than the intravenous dose of a compound of the invention, eg, Compound A' or Compound A.

[0141] It will be understood that for all embodiments described, the dosage of a compound of the invention, eg, Compound A' or Compound A, is also an effective amount of a compound of the invention, eg, Compound A' or Compound A.

[0142] In certain embodiments, an effective amount of a compound of the invention, e.g., Compound A or Compound A', when administered orally, is from about 10 to about 100 mg, from about 20 to about 750 mg, from about 50 to about 500 mg, from about 75 to about 400 mg, from about 100 to about 300 mg, from about 110 to about 290 mg, from about 120 to about 280 mg, from about 130 to about 270 mg, from about 140 to about 260 mg, from about 150 to about 250 mg, from about 160 to about 240 mg, from about 170 mg to about 230 mg, from about 180 mg to about 220 mg, from about 190 mg to about 210 mg, or about 200 mg. In another embodiment, the effective amount of a compound of the invention, e.g., Compound A or Compound A', when administered intravenously, is from about 5 to about 500 mg, from about 10 to about 400 mg, from about 25 to about 300 mg, from about 50 to about 200 mg, from about 50 to about 150 mg, from about 60 to about 140 mg, from about 70 to about 130 mg, from about 80 mg to about 120 mg, from about 90 mg to about 110 mg, or about 100 mg.

[0143] In one embodiment, the salt of the compound of the invention is a hydrochloride salt. In another embodiment, the salt of the compound of the invention is a tosylate salt. In yet another embodiment, the compound of the invention is administered orally as the free base or as the tosylate salt. In another embodiment, the compound of the invention is administered intravenously as the hydrochloride salt. In yet another embodiment, the compound of the invention is a mixed salt, for example a mixed hydrochloride and tosylate salt.

[0144] In another embodiment, a compound of the invention, eg, Compound A or Compound A', can be administered once daily, either intravenously or orally.

[0145] In some embodiments, a compound of the invention, e.g., Compound A or Compound A', can be administered for at least 3 days, at least 7 days, at least 14 days, at least 21 days, at least 30 days, or at least 60 days. For example, administration of a compound of the invention can continue for 3 to 7 days, 3 to 14 days, 3 to 21 days, 3 to 30 days, 3 to 60 days, 7 to 14 days, 7 to 21 days, 7 to 30 days, 7 to 60 days, 14 to 21 days, 14 to 30 days, 14 to 60 days, 21 to 30 days, 21 to 60 days, or 30 to 60 days.

[0146] For example, a compound of the invention, e.g., Compound A or Compound A', can be administered for 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 days, 45 days, 46 days, 47 days, 48 ​​days, 49 days, 50 days, 51 days, 52 days, 53 days, 54 days, 55 days, 56 days, 57 days, 58 days, 59 days, or 60 days.

[0147] In some embodiments, the methods include administering to the subject one or more loading doses of a compound, followed by one or more maintenance doses of the compound, hi some embodiments, the one or more loading doses can be greater than the one or more maintenance doses.

[0148] In some embodiments, administering a compound of the invention, such as Compound A or Compound A', to a subject may include administering a loading dose or doses of the compound followed by administering a maintenance dose or doses of the compound. In some embodiments, the loading dose or doses of the compound may be greater than the maintenance dose or doses of the compound. For example, the loading dose may be about 200 mg, while the maintenance dose may be about 150 mg, 100 mg, or 50 mg; or the loading dose may be about 400 mg, while the maintenance dose may be about 300 mg, 250 mg, 200 mg, 150 mg, 100 mg, or 50 mg; or the loading dose may be about 100 mg, while the maintenance dose may be about 75 mg, about 50 mg, or about 25 mg.

[0149] The loading dose of a compound of the invention and the maintenance dose of a compound of the invention can be administered by the same route or by different routes, for example, the loading dose can be administered intravenously and then the maintenance dose can be administered orally, in other embodiments, both the loading dose and the maintenance dose can be administered orally, or the loading dose and the maintenance dose can be administered intravenously.

[0150] In some embodiments, the loading dose of the compound of the invention, e.g., Compound A' or Compound A, can be an oral or intravenous dose administered twice a day, and the maintenance dose can be an oral or intravenous dose administered once a day. For example, the compound of the invention, e.g., Compound A' or Compound A, can be administered as an intravenous loading dose of 100 mg twice a day, followed by an intravenous maintenance dose of 100 mg once a day. As another example, the compound of the invention, e.g., Compound A' or Compound A, can be administered as an intravenous loading dose of 100 mg twice a day, followed by an oral maintenance dose of 300 mg once a day. For further example, the compound of the invention, e.g., Compound A' or Compound A, can be administered as an oral loading dose of 300 mg twice a day, followed by an oral maintenance dose of 300 mg once a day.

[0151] The term "treat" or "treatment" refers to the amelioration or alleviation of one or more symptoms of the disease being treated, such as a bacterial infection.

[0152] The terms "prevention," "preventing," or "prevention" refer to preventing or reducing the risk of bacterial infection.

[0153] A bacterium is "easy to make or disseminate" if it can be made or disseminated by a routine method, process, or technique, using common materials, reagents, equipment, etc. available in the art, or by a method, process, or technique, using readily accessible materials, reagents, equipment, etc., such that the bacterium can be manipulated or used by an ordinary person with little or no training in the art.

[0154] The term "moderate prevalence" refers to a prevalence of 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, or 45% or more. The term "high prevalence" refers to a prevalence of 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, or 95% or more.

[0155] The term "moderate mortality" refers to a mortality rate of 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, or 45% or more. The term "high mortality" refers to a mortality rate of 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, or 95% or more.

[0156] The terms "resistant" or "resistant" refer to antibiotic / biological standards established by the Clinical and Laboratory Standards Institute (CLSI) and / or the Food and Drug Administration (FDA).

[0157] The term "subject" includes animals that are subject to bacterial infection. Examples of subjects include animals such as farm animals (e.g., cows, pigs, horses, goats, rabbits, sheep, chickens, etc.), laboratory animals (e.g., mice, rats, monkeys, chimpanzees, etc.), pets (e.g., dogs, cats, ferrets, hamsters, etc.), birds (e.g., chickens, turkeys, ducks, geese, crows, ravens, sparrows, etc.), primates (e.g., monkeys, gorillas, chimpanzees, bonobos, and humans), and other animals (e.g., squirrels, raccoons, mice, rats, etc.). In some embodiments, the subject is a mouse or rat. In some embodiments, the subject is a cow, pig, or chicken. In some embodiments, the subject is a human.

[0158] The compounds of the present invention can be administered by any route that allows the compounds to perform their intended function, for example, to treat or prevent bacterial infection. Examples of routes include oral, intravenous, and topical. In one embodiment, the compounds of the present invention are administered orally. In another embodiment, the compounds of the present invention are administered intravenously.

[0159] The term "effective amount" refers to the amount of the compound of the present invention required to treat or prevent bacterial infection. For example, an effective amount refers to an effective level sufficient to achieve a desired therapeutic effect by cell killing and / or inhibition of cell proliferation. In some embodiments, an effective amount is an amount sufficient to eradicate the bacteria causing the infection. In some embodiments, an effective amount is the dose of the compound of the present invention, e.g., Compound A' or Compound A, administered to a subject, e.g., orally or intravenously.

[0160] The term "about" refers to a range of values ​​that may be 15%, 10%, 8%, 5%, 3%, 2%, 1%, or 0.5% greater or less than a particular value. For example, "about 10%" may be from 8.5% to 11.5%. In one embodiment, the term "about" refers to a range of values ​​that are 5% greater or less than a particular value. In another embodiment, the term "about" refers to a range of values ​​that are 2% greater or less than a particular value. In another embodiment, the term "about" refers to a range of values ​​that are 1% greater or less than a particular value.

[0161] The structures of the compounds of the present invention may contain double bonds or asymmetric carbon atoms. Such compounds may exist as racemates, racemic mixtures, individual enantiomers, individual diastereomers, diastereomeric mixtures, and cis- or trans-, or E- or Z-double bond isomers. These isomers can be obtained in substantially pure form by classical separation techniques and by stereochemically controlled synthesis. Furthermore, the structures and other compounds and moieties discussed in this invention also include all tautomers thereof.

[0162] The compounds of the present invention may be basic or acidic and can form a variety of bases with various acids or bases. Acids that can be used to prepare pharma- ceutically acceptable salts of the compounds of the present invention that are basic are those that form non-toxic acid addition salts, such as HCl, HBr, HI, nitrate, sulfate, hydrogen sulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, bitartrate, pantothenate, ascorbate, succinate, maleate, gentisic acid, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate (i.e., tosylate) and palmoate. Bases that can be used to prepare pharma-ceutically acceptable salts of the compounds of the present invention that are acidic are those that form non-toxic base salts, such as alkali metal cations (e.g., Na + and K. + ), alkaline earth metal cations (e.g. Mg ++ and Ca ++ ), and can form salts containing amines.

[0163] The compounds of the present invention that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. Acids which can be used to prepare pharma- ceutically acceptable acid addition salts of compounds of the invention that are basic in nature are those which form non-toxic acid addition salts, i.e., salts containing pharma- ceutically acceptable anions, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, hydrogen sulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate (i.e., tosylate), and palmoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoic acid)) salts. Such salts must be pharma- ceutically acceptable for administration to subjects, e.g., animals, but in practice it is often desirable to first separate the compounds of the invention from the reaction mixture as pharma- ceutically unacceptable salts, then convert the salts back to the free base compounds by simple treatment with alkaline reagents, and then convert the free bases to pharma- ceutically acceptable acid addition salts. Acid addition salts of the compounds of the invention are readily prepared by treating the compounds with substantially the same amount of a selected inorganic or organic acid in an aqueous solvent or in an organic solvent such as methanol or ethanol. Careful evaporation of the solvent readily affords the desired salt in solid form.

[0164] In one embodiment, the salt of the compound of the invention is a hydrochloride salt. In another embodiment, the salt of the compound of the invention is a tosylate salt. In yet another embodiment, the compound of the invention is administered orally as the free base or as a tosylate salt. In another embodiment, the compound of the invention is administered intravenously as a hydrochloride salt. In yet another embodiment, the compound of the invention is a mixed salt, e.g., a mixed salt of hydrochloric acid and tosylate.

[0165] It will be appreciated that wherever values ​​and ranges are given herein, e.g., age of a subject population, dosage, duration, etc., all values ​​and ranges encompassed by these values ​​and ranges are intended to be within the scope of the invention. Moreover, all values ​​within these values ​​and ranges may be the upper or lower limits of a range.

[0166] The compounds of the present invention can be synthesized by using art-recognized techniques, such as those described in U.S. Patent Nos. 6,846,939 and 7,553,828, and U.S. Patent Publication No. 20080287401, each of which is incorporated herein by reference in its entirety. The compounds thus obtained can be further purified, for example, by flash column chromatography, high performance liquid chromatography, crystallization, or any known purification method.

[0167] In certain embodiments, compounds of the present invention can be synthesized according to the synthetic scheme described in U.S. Patent Publication No. 20080287401, shown below:

[0168] [ka]

[0169] In certain embodiments, the compounds of the present invention can be purified by chromatography, which involves injecting a low pH aqueous solution of the compound into a polar organic solvent gradient HPLC and combining the product fractions. Selection of an appropriate acidic mobile phase enhances process stability and selectivity. Organic and mineral acid mobile phases are effective in separating by-products, including epimeric impurities and closely eluted by-products, by pH control or acid selection. Acidic mobile phases also protect the compounds from oxidative degradation. For example, the pH of the low pH solution is about 2-3. Examples of solutions that can be used include 0.1% methanesulfonic acid in water and 0.1% trifluoroacetic acid in water. An isocratic gradient of 94% aqueous solution and 6% acetonitrile or another polar organic solvent can be used to purify the compound from epimers and closely eluted by-products. The resulting aqueous product fractions can be combined and the pH adjusted to about 4.0-04.5 with a base (e.g., NaOH). Hydrophobic impurities and oxidative decomposition products of the compound can be removed by washing the aqueous solution with a non-polar organic solvent (e.g., CH2Cl2). The organic layer can be discarded and the aqueous layers can be combined and saved. Of note, an organic solvent such as methylene chloride can be used to selectively remove late eluting hydrophobic impurities such as the 4-carbonyl by-product, as well as oxidative decomposition products from the acidic aqueous solution of the compound. The pH of the combined aqueous layers is adjusted to a neutral pH (e.g., about 7.5 to about 8.5). The pH can be adjusted by the addition of a base such as NaOH. The neutral solution can then be washed with a non-polar solvent, e.g., methylene chloride. Of note, selective pH adjustment to a neutral pH range can allow extraction of the compound into the organic solvent while retaining the undesired β-epimer and by-products in the aqueous layer.

[0170] The reagents that can be used in the synthetic routes described in the above patents can be, for example, solvents, reagents, catalysts, and protecting group and deprotecting group reagents. The synthetic routes can include additional steps before or after the steps specified therein to add or remove appropriate protecting groups to ultimately allow synthesis of the desired tetracycline compound. In addition, various synthetic steps can be performed in different orders to provide the desired compounds. For example, the compounds can be further modified by conventional chemical transformations to produce the compounds of the present invention. Synthetic chemistry transformations and protecting group methodology (protection and deprotection) are known in the art and may be found, for example, in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); TW Greene and PGM Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley and Sons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995).

[0171] The synthetic routes described in the above patents are used for illustrative purposes only. It will be appreciated by those skilled in the art that in light of the schemes and examples described in the above patents, all compounds of the present invention can be made by similar methods well known to those skilled in the art.

[0172] The efficacy of the compounds of the present invention in treating or preventing bacterial infections can be evaluated using common methods known in the art. In some embodiments, efficacy can be measured by a minimum inhibitory concentration (MIC) assay. For example, the compounds of the present invention are serially diluted and then added to a growth medium, such as a cation-adjusted Mueller-Hinton Broth (CAMHB) for bacterial cultures. The minimum concentration of the compounds of the present invention that inhibits bacterial growth by 50% or 90% (i.e., MIC 50 or MIC 90 ) and, if necessary, the MICs of other antibiotics. 50 or MIC 90 In another embodiment, efficacy can be measured by in vivo assays known in the art (e.g., animal studies). For example, the compounds of the present invention are administered in decreasing amounts to laboratory animals (e.g., mice and rats). The minimum amount of the compounds of the present invention that treats the laboratory animals (e.g., improves symptoms of bacterial infection, extends the life of the animals, and allows the animals to survive bacterial infection) or prevents the laboratory animals from being infected with bacteria or from developing symptoms of infection is determined, and if necessary, compared to the minimum amount of other antibiotics that achieve the same results.

[0173] The present invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of the present invention (e.g., Compound A) or a salt thereof, and optionally a pharma- ceutically acceptable carrier. In another embodiment, the present invention relates to a pharmaceutical composition comprising about 10 to about 1000 mg of a compound of the present invention (e.g., Compound A or Compound A') or a salt thereof, and a pharma- ceutically acceptable carrier. In yet another embodiment, the pharma- ceutically acceptable carrier satisfies the conditions for oral administration. In yet another embodiment, the compound of the present invention is a free base or a tosylate salt.

[0174] In yet another embodiment, the composition contains about 20 to about 750 mg, about 50 to about 500 mg, about 75 to about 400 mg, about 100 to about 300 mg, about 110 to about 290 mg, about 120 to about 280 mg, about 130 to about 270 mg, about 140 to about 260 mg, about 20 to about 150 to about 250 mg, about 160 to about 240 mg, about 170 mg to about 230 mg, about 180 mg to about 220 mg, about 190 mg to about 210 mg, or about 200 mg of a compound of the present invention (e.g., Compound A or Compound A') or a salt thereof.

[0175] In another embodiment, the present invention also relates to a pharmaceutical composition containing about 5 to about 500 mg of a compound of the present invention (e.g., Compound A or Compound A') or a salt thereof, and a pharma- ceutically acceptable base suitable for intravenous administration. In yet another embodiment, the composition contains about 10 to about 400 mg, about 25 to about 300 mg, about 50 to about 200 mg, about 50 to about 150 mg, about 60 to about 140 mg, about 70 mg to about 130 mg, about 80 mg to about 120 mg, about 90 mg to about 110 mg, or about 100 mg of a compound of the present invention (e.g., Compound A or Compound A') or a salt thereof.

[0176] The term "pharmaceutical acceptable carrier" includes a substance that can be administered together with a compound of the present invention (e.g., Compound A or Compound A') and that allows the compound to perform its intended function, for example, to treat or prevent bacterial infection. Suitable pharmaceutical acceptable carriers include, but are not limited to, water, saline, alcohol, vegetable oils, polyethylene glycol, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, flavor oils, fatty acid monoglycerides and diglycerides, petroethral fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone, and the like. The pharmaceutical preparations are sterilized and, if necessary, can be mixed with auxiliary agents that do not adversely react with the compound of the present invention, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts that affect osmotic pressure, buffers, colorants, flavors and / or aromatics.

[0177] The compound of the present invention and its pharmaceutically acceptable salts can be administered by oral, parenteral or topical route.Generally, these compounds are most preferably administered in effective dosages according to the weight and condition of the subject to be treated and the individual administration route selected.Variations may occur according to the species of the subject to be treated and the individual response to drugs, as well as the type of pharmaceutical preparation selected, and the duration and interval of carrying out such administration.

[0178] The compounds and pharmaceutical compositions of the present invention can be administered alone or in combination with other known compositions to treat a tetracycline-responsive condition in a subject. The term "in combination with" a known composition is intended to include administering the composition of the present invention and the known composition simultaneously, administering the composition of the present invention first, followed by the known composition, and administering the known composition first, followed by the composition of the present invention. Any therapeutic composition known in the art for treating a tetracycline-responsive condition can be used in the methods of the present invention.

[0179] The compounds and pharmaceutical compositions of the present invention can be administered by any of the above routes, either alone or in combination with pharma- ceutically acceptable carriers or diluents, and can be administered in single or multiple doses. For example, the compounds of the present invention can be effectively administered in a variety of different dosage forms, i.e., the compounds of the present invention can be combined with various pharma- ceutical acceptable inert carriers to form tablets, capsules, lozenges, troches, hard candies, powders, sprays, creams, salves, suppositories, jellies, gels, pastes, lotions, ointments, aqueous suspensions, injectables, elixirs, syrups, and the like. Such carriers include solid excipients or bulking agents, sterile aqueous solvents, and various non-toxic organic solvents. Furthermore, oral pharmaceutical compositions can be suitably sweetened and / or flavored. In general, the therapeutically effective compounds of the present invention are included in such dosage forms at concentration levels ranging from about 5.0% to about 70% by weight.

[0180] For oral administration, tablets containing various additives such as microcrystalline cellulose, sodium citrate, calcium carbonate, dicalcium phosphate and glycine can be used with granulation binders such as polyvinylpyrrolidone, sucrose, gelatin, and gum arabic, in conjunction with various disintegrants such as starch (preferably corn, potato, or tapioca starch), alginic acid, and certain complex silicates. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate, and talc are often very useful for tabletting purposes. Solid compositions of a similar type can also be used as fillers in gelatin capsules; preferred materials in this connection also include lactose or milk sugar as well as high molecular weight polyethylene glycols.

[0181] When aqueous suspensions and / or elixirs are desired for oral administration, the active ingredient can be combined with diluents such as water, ethanol, polyethylene glycol, glycerin, and various similar combinations thereof, along with various sweetening or flavoring agents, coloring substances or dyes, and, if desired, further emulsifying and / or suspending agents.

[0182] For parenteral administration (including intraperitoneal, subcutaneous, intravenous, intradermal, or intramuscular injection), solutions of the compounds of the present invention in sesame or peanut oil or in aqueous propylene glycol can be used. Aqueous solutions should be appropriately buffered as necessary, and the liquid diluent should be rendered isotonic to begin with. These solutions are suitable for intravenous injection. Oily solutions are suitable for intra-articular, intramuscular, and subcutaneous injection. Preparation of all these solutions under sterile conditions is easily accomplished by standard pharmaceutical techniques well known to those skilled in the art. For parenteral application, examples of suitable formulations include solutions, preferably oily or aqueous solutions, as well as suspensions, emulsions, or placements, including suppositories. The therapeutic compounds may also be formulated under sterile conditions in multiple or single dose forms dispersed in a liquid base, such as sterile saline or 5% saline dextrose, commonly used with injections.

[0183] For enteric application, particularly suitable are tablets, dragees, or capsules having talc and / or carbohydrate-based binders, preferably lactose and / or corn starch and / or potato starch.Syrups, elixirs, etc. using sweetening agents can be used.Sustained-release compositions can be formulated, including compositions in which the active ingredient is protected by a gradually degradable coating, for example by microencapsulation, multiple coatings, etc.

[0184] Illustration of the invention EXAMPLES

[0185] [ka]

[0186] Minocycline hydrochloride was dissolved in methylsulfonic acid or hydrofluoric acid along with methylsulfonic anhydride. N-hydroxymethylphthalimide was added to the reaction mixture. The mixture was stirred at 20-35°C until the reaction was complete. When the acid solution was added to an ice / water mixture, the triflate salt immediately precipitated and was collected by filtration. This salt was redissolved in acetone and brought to neutral pH with base. The product was precipitated by the addition of water. The product was isolated as a mixture of bis and tris alkylated products. The isolated material from this reaction was concentrated to the desired bis ratio (90%).

[0187] The solid was suspended in EtOH. Aminolysis was carried out with methylamine. As the reaction proceeded, the phthalamide by-product precipitated and was removed by filtration. Addition of approximately 1.5 volumes of t-butyl methyl ether to the reaction mixture precipitated a pale yellow solid product that was collected by simple filtration, leaving many minor impurities and the methylamine reagent in solution. The compound was further purified by reslurrying in methanol.

[0188] The free base of compound 4 was transferred to a hydrogenation reactor containing methanol and an aldehyde. Deactivated Pd / C catalyst was added and the vessel was pressurized with hydrogen gas. The reaction mixture was hydrogenated for about 24 hours under about 30 Psi of hydrogen pressure. Upon completion of the conversion of compound 4 to 1, the solution was filtered and washed through a Celite pad. At this point, the reaction mixture contained only a small amount of the βC-4 epimer, about 3-7%.

[0189] The product (1) was worked up and selectively isolated from impurities. The pH of the solution was adjusted to about 4.5 using concentrated HCl, and the solution was washed with dichloromethane. Sulfite was added to the aqueous layer, and the product was extracted with dichloromethane at about pH 7-8 to selectively recover the desired epimeric product (e.g., a). The dichloromethane layers were combined and concentrated, and 2 L of n-heptane was added to precipitate the product. The crude product was further purified by repeating the workup step to dissolve with or without tert-butyl methyl ether.

[0190] 9-(2',2'-Dimethylpropylaminomethyl)-minocycline dihydrochloride (200 mg, 1 equiv.), DMF, and trimethylacetaldehyde (45 μL, 1 equiv.) were combined in a 40 mL flask and stirred. Triethylamine (150 μL, 3 equiv.) was then added. After stirring at room temperature for a few minutes, NaBH(OAc)3 (175 mg, 2 equiv.) and InCl3 (9 mg, 0.1 equiv.) were added. After 1 h, the reaction was clear and red in color. The reaction was quenched with methanol and the solvent was removed to give compound A. EXAMPLES

[0191] Crude 9-(2',2'-dimethylpropylaminomethyl)-minocycline free base (40 g) was dissolved in 150 mL of Buffer A (0.1% methanesulfonic acid (MSA) in water) and the pH was adjusted to 2-3 with MSA. The solution was filtered and injected into the HPLC, and the product was eluted with an isocratic gradient of 94% Buffer A and 6% acetonitrile. Collection of product fractions was initiated when the product peak was detected. Each fraction was analyzed, and an acceptance criterion of AUC of the main peak >80% was used for early product fractions. When fractions were combined, the levels of impurities, and the relative concentrations of the pooled fractions, were included in the selection criteria for meeting the final product specifications. A 10% aqueous solution of sodium sulfite was added to the product fractions equal to 10% of the original volume of the collected fractions.

[0192] A product fraction of 3.5 liters (containing sodium sulfite) was collected and the pH was adjusted to 4.0-4.5 with sodium hydroxide solution. The aqueous solution was washed with 2 liters of dichloromethane and the organic layer was separated and discarded. The pH of the aqueous layer was adjusted to 7.5-8.5 with sodium hydroxide and the product was extracted four times with 2.4 liters of dichloromethane. Before each extraction the pH was adjusted to 7.5-8.5 with sodium hydroxide.

[0193] The four dichloromethane layers were combined and concentrated to approximately 200 mL, which was then slowly (over approximately 10 min) added to vigorously stirred n-heptane (2.5 L). The suspension was stirred at room temperature for approximately 10 min and slowly diluted (over 5 min) with 1.5 L of n-heptane. The slurry was cooled to 0-5° C. and stirred for 1-2 h. The suspended solids were filtered and washed with three 150 mL portions of n-heptane. The product was dried under vacuum at 40° C. for at least 24 h until a constant weight was reached and all residual solvent levels were within the specified range. Approximately 13.6 g of 9-(2′,2′-dimethylpropylaminomethyl)-minocycline free base was isolated as a yellow solid. The remaining product was similarly isolated, weighing 1.64 g. EXAMPLES

[0194] Bacterial inocula were prepared by suspending colonies from 18-24 h B. anthracis, B. pseudomallei, or B. mallei plates or 42-48 h F. tularensis or Y. pestis plates incubated at 35°C in cation-adjusted Mueller Hinton broth (CAMHB). Sheep blood agar (SBA) plates were used for B. anthracis and Y. pestis, chocolate agar plates for F. tularensis, and trypticase soy agar (TSA) plates for B. pseudomallei and B. mallei. Suspension cultures were plated at a bacterial cell density of 10 5 The OD 600 The conversion factors used for each pathogen were: B. anthracis (3.82 x 10 7 CFU / mL / OD), B. mallei and B. pseudomallei (5.0 x 10 8 CFU / mL / OD), Y. pestis (5.34 x 10 8 CFU / mL / OD), and F. tularensis (3.89 x 10 10 CFU / mL / OD) was approximately 5x10 4 For a final inoculum of CFU / well, 50 μL of the adjusted dilution was added to each well of a 96-well plate.

[0195] The inoculated 96-well plates were incubated at 35°C. Antibiotics were serially diluted 2-fold in 50 μL of CAMHB and added to each well of the plate. For all steps with F. tularensis, CAMHB was supplemented with 2% Isovitalex (Becton Dickinson). Antibiotics ranged from 8-0.0039 μg / ml, or 64-0.03125 μg / ml, based on a final well volume of 100 μL after inoculation. MICs were determined by microdilution in 96-well plates according to CLSI guidelines. MICs were determined visually at 18-24 h or 42-48 h (for F. tularensis and Y. pestis) and also by reading plates at 600 nm (SpectroMax M2, Molecular Devices). Quality control of antibiotic stock solutions was established by using E. coli ATCC 25922, P. aeruginosa ATCC 27853, and S. aureus ATCC 29213. Inocula of these control bacteria were prepared as described above from 18-24 hour SBA plates. Conversion factors were E. coli (6.83x10 8 CFU / mL / OD), P. aeruginosa (5.74x10 10 CFU / mL / OD), and Staphylococcus aureus (2.07xl0 10 CFU / mL / OD).

[0196] A stock solution of 5.15 mg / ml of Compound A was made in 100% DMSO. Prior to use, the stock solution was diluted in CAMHB and then added to the 96-well plate as described above. The control antibiotics, ciprofloxacin, ceftazidime, and azithromycin, were all purchased from USP, made into 5.15 mg / ml stock solutions according to CLSI M 100-S 18 Table 4 guidelines, and stored at -70°C until use. EXAMPLES

[0197] Compound A is highly effective against strains of Bacillus anthracis and B. mallei and also has significant activity against Yersinia pestis and Franciscella tularensis, with excellent MICs against these pathogens. 50 and MIC 90 The MIC values ​​for compound A were comparable to those of doxycycline, which is currently recommended for the treatment of disease caused by these pathogens. However, unlike doxycycline, compound A is expected to be active against strains that possess either efflux or ribosomal resistance mechanisms.

[0198] [Table 1] EXAMPLES

[0199] The in vitro activity of Compound A was evaluated in an independent laboratory using bacteria isolated from clinical specimens. For all anaerobic or facultative anaerobic organisms, MICs were determined using cation-adjusted Mueller Hinton Broth (CAMHB, DIFCO Lot# 5230237) by the broth microdilution method recommended by the Clinical & Laboratory Standards Institute (CLSI, M7-A7, 2006). CAMHB was supplemented with 3-5% horse lysed blood (Hemostat Lot# H06036) when testing for difficult-to-culture organisms such as streptococci. Haemophilus test medium (HTM) was used to test for Haemophilus influenzae. Simple Mueller Hinton agar (Difco Lots 1303004 & 5011641) was used to test the majority of isolates, and 5% sheep blood (Hema Resource Lot # 1127-100140-04) was used to test for streptococci, as recommended by the CLSI document M7-A7, Selected microbial groups may also be tested by agar dilution using methods specified by the MIC Guidelines in 2006. All cultures used were less than 12 hours old at the time of preparation of the MIC trays.

[0200] All anaerobic strains were tested by broth microdilution and agar dilution using the methods specified by CLSI (M11-A6, 2004). All anaerobic strains were tested using Brucella broth (BBL Lot #5227153) supplemented with 5 μg / ml hemin (Sigma Lot #89K0914), 1 μg / ml vitamin K1 (Sigma Lot #120K1413), and 5% lysed horse blood (Hemostat Lot #H06036). In addition, all strains were tested by agar dilution using Brucella agar (BBL Lot #6160970) supplemented with 5 μg / ml hemin, 1 μg / ml vitamin K1, and 5% lysed sheep blood (Hema Resource Lot # 1127-100140-04). Fresh cultures less than 12 hours old were used to prepare MIC trays. Agar dilution plates were poured on the day of testing.

[0201] Compound A was active against almost all clinical isolates tested (Table 2), including isolates resistant to current tetracycline antibiotics such as doxycycline (Table 2). Compound A was also active against pathogens expected to be found in ABSSSI and CABP, regardless of resistance to commonly used antibiotics, including tetracycline. The in vitro activity of Compound A was not affected by serum or pulmonary surfactant, an important property consistent with its potential utility in lower respiratory tract infections.

[0202] [Table 2] a ND - not tested. bMICs determined by broth microdilution using fresh media. Data taken from Brown, S., and MM Traczewski, 2007. MK-2764: In vitro Spectrium of Activity, Confirmation of Disk Mass, Agar Dilution Validation and Short Term Stability using Fresh Media, The Clinical Microbiology Institute Report, Wilsonville, OR. EXAMPLES

[0203] We demonstrated the in vivo activity of Compound A in multiple animal models of infection with a variety of pathogens. As shown in Table 3, Compound A was generally as potent and effective as, or more potent than, minocycline, vancomycin, and linezolid.

[0204] [Table 3] a PD 50 (50% protective dose) is defined as the dose required to achieve 50% survival. b ED 50 The (50% effective dose) is defined as the dose required to achieve a 2 log10 reduction in bacterial burden (cfu / g) in target organs versus untreated controls. EXAMPLES

[0205] The in vivo activity of compound A (omadacycline) was investigated in a lethal Y. pestis postexposure prophylaxis (PEP) infection model. Six- to eight-week-old female BALB / c mice were infected with LD 50Mice were infected with Y. pestis (CO92) at 29.9. Ten mice were used per group. Compound A was administered intraperitoneally at doses of 5, 10, 20 and 40 mg / kg 24 hours after infection, every 12 hours for 7 days. As positive controls, doxycycline was used at doses of 5, 10, 20 and 40 mg / kg, and ciprofloxacin at a dose of 15 mg / kg. Mice were followed up for 14 days after infection, and the survival percentage was determined.

[0206] The results are shown in Figure 1 and show that Compound A at 40 mg / kg is more effective than doxycycline at 40 mg / kg and at least as effective as ciprofloxacin at 15 mg / kg in treating Y. pestis infection for at least 14 days post-infection. In this experiment, the MIC of Compound A was 1 μg / mL, doxycycline was 0.5 μg / mL, and ciprofloxacin was 0.06 μg / mL. The PD of Compound A was 0.5 μg / mL, doxycycline was 0.5 μg / mL, and ciprofloxacin was 0.06 μg / mL. 50 The PD of doxycycline was 23.5 (range 20.1–27.0 mg / kg). 50 was 29.7 (range 20.7-38.8). EXAMPLES

[0207] The in vivo activity of compound A (omadacycline) was examined in a lethal B. anthracis post-exposure prophylaxis (PEP) infection model. Six- to eight-week-old female BALB / c mice were infected with LD 50 Mice were infected with B. anthracis AMES strain 30.5. Ten mice were used per group. Compound A was administered intraperitoneally 24 hours after infection at doses of 0.75, 2.5, 7.5, and 15 mg / kg every 12 hours for 14 days. As positive controls, doxycycline was used at doses of 0.75, 2.5, 7.5, and 15 mg / kg, and ciprofloxacin was used at a dose of 30 mg / kg. Mice were followed up for 41 days after infection to determine the survival percentage.

[0208] The results, shown in Figure 2, indicate that compound A is as effective as doxycycline or ciprofloxacin in treating B. anthracis infection for at least 41 days post-infection. In this experiment, the MIC of compound A was <0.03 μg / mL, whereas the MICs of doxycycline and ciprofloxacin were both 0.03 μg / mL. The PD of compound A was 50 The PD of doxycycline was 0.8 (range 0.6–1.1 mg / kg). 50 was 2.0 (range 1.4-2.6). EXAMPLES

[0209] The in vivo activity of compound A (omadacycline) was investigated in a lethal B. mallei post-exposure prophylaxis (PEP) infection model. Six- to eight-week-old female BALB / c mice were infected with LD 50 Mice were infected with B. mallei (China 7) strain 59.6. Ten mice were used per group. Compound A was administered intraperitoneally at doses of 0.75, 2.5, 7.5 and 15 mg / kg 24 hours after infection, every 12 hours for 21 days. As positive controls, doxycycline was used at concentrations of 0.75, 2.5, 7.5 and 15 mg / kg, and azithromycin at a concentration of 15 mg / kg. Mice were followed up for 55 days after infection, and the survival percentage was determined.

[0210] The results are shown in Figure 3 and indicate that compound A is as effective as doxycycline or azithromycin in treating B. mallei infections. In this experiment, the MIC of compound A was 0.25 μg / mL, the MIC of doxycycline was 0.06 μg / mL, and the MIC of azithromycin was 0.5 μg / mL. The PD of compound A was 50 The PD of doxycycline was <0.75 mg / kg. 50 The dose was <0.75 mg / kg. All but one animal in the treated groups survived all treatments. No animals in the control groups survived.

[0211] equivalent Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments and methods described herein. Such equivalents are intended to be within the scope of the present invention. All patents, patent applications, and literature references cited herein are expressly incorporated by reference.

Claims

1. 1. A method of treating a bacterial infection in a subject in need thereof, comprising administering to the subject an effective amount of a compound or a salt thereof at a dose of from about 10 mg to about 1000 mg, the compound having the structural formula: 【Chemistry 1】 wherein the bacterial infection is caused by a bacteria that can be used as a biological weapon, thereby treating the infection in the subject.

2. Compound A, wherein the compound has the following structural formula: 【Chemistry 2】 The method of claim 1, wherein

3. 10. The method of claim 1, wherein the bacteria is resistant to an antibiotic commonly used to treat bacterial infections.

4. The method of claim 1, wherein the bacteria is in the form of a powder or aerosol.

5. The method of claim 1 , wherein the bacterium is capable of forming spores.

6. Bacteria: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. cholerae), Salmonella (e.g., S. bongori and S. enterica); and Bacteria belonging to the diarrheagenic strains of Escherichia coli (E. coli), The method of claim 1 , wherein the compound is selected from the group comprising:

7. Bacteria: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (e.g., B. anthracis, including multidrug-resistant (MDR) anthrax strains); The method of claim 6, wherein the compound is selected from the group comprising:

8. Bacteria: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. cholerae), Salmonella (e.g., S. bongori and S. enterica); and Bacteria belonging to the diarrheagenic strains of Escherichia coli (E. coli), The method of claim 1 or 2, wherein the compound is selected from the group consisting of:

9. Bacteria: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (e.g., B. anthracis, including multidrug-resistant (MDR) anthrax strains); The method of claim 8, wherein the compound is selected from the group consisting of:

10. 2. The method of claim 1, wherein the bacterium is selected from the group consisting of Yersinia pestis, Burkholderia mallei, and B. anthracis.

11. 10. The method of claim 1, wherein the compound is administered once daily or twice daily.

12. 10. The method of claim 1, wherein the compound is administered intravenously or orally.

13. 10. The method of claim 1, wherein the compound is administered intravenously at a dose of from about 50 mg to about 200 mg.

14. 14. The method of claim 13, wherein the compound is administered intravenously at a dose of about 100 mg.

15. 10. The method of claim 1, wherein the compound is orally administered at a dose of from about 100 mg to about 300 mg.

16. 16. The method of claim 15, wherein the compound is administered orally at a dose of about 300 mg.

17. 10. The method of claim 1, wherein the method comprises administering the compound for at least 3 days, at least 7 days, at least 14 days, at least 21 days, at least 30 days, or at least 60 days.

18. 20. The method of claim 17, wherein the method comprises administering the compound for about 30 days.

19. 10. The method of claim 1, wherein the method comprises administering to the subject one or more loading doses of the compound, followed by one or more maintenance doses of the compound.

20. 20. The method of claim 19, wherein the one or more loading doses are greater than the one or more maintenance doses.

21. 20. The method of claim 19, wherein the loading dose is intravenous and the maintenance dose is oral.

22. The method of claim 1 , wherein the subject is a human.

23. 1. A method of preventing a bacterial infection in a subject in need thereof, comprising administering to the subject an effective amount of a compound or a salt thereof at a dose of from about 10 mg to about 1000 mg, the compound having the following structural formula: 【Chemistry 3】 wherein the bacterial infection is caused by a bacteria that may be used as a biological weapon, thereby preventing the infection in the subject.

24. The compound is Compound A, which has the following structural formula: 【Chemistry 4】 24. The method of claim 23.

25. 24. The method of claim 23, wherein the bacteria is resistant to an antibiotic commonly used to treat bacterial infections.

26. 24. The method of claim 23, wherein the bacteria is in the form of a powder or aerosol.

27. 24. The method of claim 23, wherein the bacterium is capable of forming spores.

28. Bacteria: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. cholerae), Salmonella (S. bongori and S. enterica); and Bacteria belonging to the diarrheagenic strains of Escherichia coli (E. coli), 24. The method of claim 23, wherein the compound is selected from the group comprising:

29. Bacteria: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (e.g., B. anthracis, including multidrug-resistant (MDR) anthrax strains); 24. The method of claim 23, wherein the compound is selected from the group comprising:

30. Bacteria: Bacteria belonging to the following species: Franciscella tularensis, Clostridium botulinum, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii, Chlamydia psittaci, Clostridium perfringens, Rickettsia prowazekii, Campylobacter jejuni, Yersinia enterocolitica, Listeria monocytogenes; Bacillus (e.g., B. anthracis, including multidrug resistant (MDR) strains of B. anthracis), Brucella (e.g., B. abortus, B. canis, B. ceti, B. inopinata, B. melitensis, B. microti, B. neotomae, B. ovis, B. pinnipedialis, B. suis), Shigella (e.g., S. boydii, S. dysenteriae, S. flexneri, and S. sonnei), Vibrio (e.g., V. cholerae, V. cholerae), Salmonella (e.g., S. bongori and S. enterica); and Bacteria belonging to the diarrheagenic strains of Escherichia coli (E. coli), 25. The method of claim 23 or 24, selected from the group consisting of:

31. Bacteria: Bacteria belonging to the species Franciscella tularensis, Yersinia pestis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia prowazekii; and Bacteria belonging to the genus Bacillus (e.g., B. anthracis, including multidrug-resistant (MDR) anthrax strains); 31. The method of claim 30, wherein the compound is selected from the group consisting of:

32. 24. The method of claim 23, wherein the bacterium is selected from the group consisting of Yersinia pestis, Burkholderia mallei, and B. anthracis.

33. 24. The method of claim 23, wherein the compound is administered once daily or twice daily.

34. 24. The method of claim 23, wherein the compound is administered intravenously or orally.

35. 24. The method of claim 23, wherein the compound is administered intravenously at a dose of from about 50 mg to about 200 mg.

36. 36. The method of claim 35, wherein the compound is administered intravenously at a dose of about 100 mg.

37. 24. The method of claim 23, wherein the compound is orally administered at a dose of from about 100 mg to about 300 mg.

38. 38. The method of claim 37, wherein the compound is administered orally at a dose of about 300 mg.

39. 24. The method of claim 23, wherein the method comprises administering the compound for at least 3 days, at least 7 days, at least 14 days, at least 21 days, at least 30 days, or at least 60 days.

40. 40. The method of claim 39, wherein the method comprises administering the compound for about 30 days or about 60 days.

41. 24. The method of claim 23, wherein the method comprises administering to the subject one or more loading doses of the compound, followed by one or more maintenance doses of the compound.

42. 42. The method of claim 41, wherein the one or more loading doses are greater than the one or more maintenance doses.

43. 42. The method of claim 41, wherein the loading dose is intravenous and the maintenance dose is oral.

44. 24. The method of claim 23, wherein the subject is a human.

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