Compounds and Methods for Treating Chronic

Microbial Infections

CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application serial number 62/773,703, filed November 30, 2018, which is hereby incorporated by reference in its entirety.

GOVERNMENT SUPPORT

This invention was made with government support under Grant No. R01AI093459 and Grant No. R01AI125362 awarded by the National Institutes of Health. The government has certain rights in the invention.

STATEMENT REGARDING PRIOR DISCLOSURE BY AN INVENTOR OR A JOINT

INVENTOR UNDER 37 CFR 1.77(b)(6)

On November 30, 2017, Lizbeth K. Hedstrom publicly disclosed a poster entitled “New Strategy to Treat Chronic Infections.”

BACKGROUND

Most recurrent and difficult to treat infections are caused by bacteria that are susceptible to commonly used antibiotics. Examples of common chronic bacterial infections include, but are not limited to, surgical site infections, wound infections, prosthetic joint infections, osteomyelitis, and endocarditis (caused by Staphylococcus aureus ); Lyme disease (caused by bacteria in genus Borrelia , such as B. burgdorferi ); and tuberculosis (caused by Mycobacterium tuberculosis).

These infections persist because antibiotics are only effective against actively growing bacteria. However, a small number of bacteria in an infection are quiescent, and therefore survive treatment. These quiescent cells are known as“persisters.” Persister cells are hypothesized to play an important role in the emergence of antibiotic resistance. While some anti-persister compounds have been described in the art, none has reached the clinic.

There is a need for compounds that can be used to treat recurrent or chronic bacterial infections, either alone or in combination with commonly used antibiotics. BRIEF SUMMARY

In certain embodiments, the invention relates to a compound having the following structure:

, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a compound having a structure selected from the group consisting of:

In certain embodiments, the invention relates to a pharmaceutical composition, comprising a pharmaceutically acceptable carrier, adjuvant, or vehicle; and a compound having the following structure:

, or a pharmaceutically acceptable salt thereof. In certain embodiments, the invention relates to a pharmaceutical composition, comprising a pharmaceutically acceptable carrier, adjuvant, or vehicle; and a compound having a structure selected from the group consisting of:

pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a pharmaceutical composition, comprising

(a) a compound having the following structure:

, or a pharmaceutically acceptable salt thereof;

(b) an antibacterial agent; and

(c) a pharmaceutically acceptable carrier, adjuvant, or vehicle. In certain embodiments, the invention relates to a pharmaceutical composition, comprising

(a) a compound having a structure selected from the group consisting of:

pharmaceutically acceptable salt thereof;

(b) an antibacterial agent; and

(c) a pharmaceutically acceptable carrier, adjuvant, or vehicle.

In certain embodiments, the invention relates to a pharmaceutical composition, comprising

(a) a compound selected from the group consisting of:

(b) an antibacterial agent; and

(c) a pharmaceutically acceptable carrier, adjuvant, or vehicle.

In certain embodiments, the invention relates to a method of killing or inhibiting the growth of a microbe, comprising the step of contacting said microbe with an effective amount of a compound having the following structure:

, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a method of treating or preventing a microbial infection in a mammal, comprising the step of administering to a mammal in need thereof a therapeutically effective amount of a compound having the following structure: , or a pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a method of treating or preventing a parasitic infection in a mammal comprising the step of administering to a mammal in need thereof a therapeutically effective amount of a compound having the following structure:

, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a method of killing or inhibiting the growth of a microbe, comprising the step of contacting said microbe with an effective amount of a compound having a structure selected from the group consisting of:

pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a method of treating or preventing a microbial infection in a mammal, comprising the step of administering to a mammal in need thereof a therapeutically effective amount of a compound having a structure selected from the group consisting of:

In certain embodiments, the invention relates to a method of treating or preventing a parasitic infection in a mammal comprising the step of administering to a mammal in need thereof a therapeutically effective amount of a compound having a structure selected from the group consisting of:

In certain embodiments, the invention relates to a method of treating or preventing a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of a compound having the following structure:

, or a pharmaceutically acceptable salt thereof; and

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are effective.

In certain embodiments, the invention relates to a method of treating or preventing a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of a compound having a structure selected from the group consisting of:

pharmaceutically acceptable salt thereof; and

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are effective. In certain embodiments, the invention relates to a method of treating a recurrent or persistent bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of any one of the compounds described in US Patent Application Publication No. US 2010/0022547, US Patent No. 8,969,342, or US Patent No. 9,447,134, each of which is incorporated by reference in its entirety; and

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are therapeutically effective.

In certain embodiments, the invention relates to a method of treating a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of any one of the compounds described in US Patent Application Publication No. US 2010/0022547, US Patent No. 8,969,342, or US Patent No. 9,447,134, each of which is incorporated by reference in its entirety; and

(b) a second amount of an antibacterial agent,

wherein,

taken together, the first amount and the second amount are therapeutically effective; and

the bacterial infection does not persist or recur after treatment.

In certain embodiments, the invention relates to a method of preventing the recurrence of a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of any one of the compounds described in US Patent Application Publication No. US 2010/0022547, US Patent No. 8,969,342, or US Patent No. 9,447,134, each of which is incorporated by reference in its entirety; and

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are effective to prevent the recurrence of the bacterial infection in the mammal. In certain embodiments, the invention relates to a method of treating a recurrent or persistent bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of a compound selected from the group consisting of:

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are therapeutically effective.

In certain embodiments, the invention relates to a method of treating a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of a compound selected from the group consisting of:

(b) a second amount of an antibacterial agent,

wherein,

taken together, the first amount and the second amount are therapeutically effective; the bacterial infection does not persist or recur after treatment.

In certain embodiments, the invention relates to a method of preventing the recurrence of a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of a compound selected from the group consisting of:

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are effective to prevent the recurrence of the bacterial infection in the mammal.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1A is the structure of a compound useful in the compositions and methods described herein. Figure IB is the structure of a compound of the invention, which also is useful in the compositions and methods described herein.

Figure 1C is the structure of a compound useful in the compositions and methods described herein.

Figure ID is the structure of a compound useful in the compositions and methods described herein.

Figure 2 is a graph showing that a population of cells survives treatment with conventional antibiotics alone. Stationary phase Staph bacteria were treated with ciprofloxacin (Cipro), kanamycin (Kan) or oxacillin (Oxa) at lOxMIC in 50% small molecule fraction (SMF) of human serum.

Figure 3A is a graph showing that compound P226 induces premature bacterial growth (S. aureus ), thereby sensitizing bacteria to commonly used antibiotics. Ctrl, RPMI media; Serum, 50% small molecule fraction (SMF) of human serum in RPMI; + P226 (5 mM = 4xMIC).

Figure 3B is a bar graph showing stationary phase S. aureus cells were treated with P226 (4xMIC), ciprofloxacin (Cipro), kanamycin (Kan), or oxacillin (Oxa) at lOxMIC in 50% SMF of human serum. Bacteria (CFUs) were measured after 18 h.

Figure 3C is a photograph showing that small colonies were not observed in P226- treated cells.

DETAILED DESCRIPTION

Overview

One aspect of the invention relates to compounds and pharmaceutically acceptable salts thereof that initiate, induce, or accelerate growth of a bacterium, for example, a quiescent bacterium. In some embodiments, the compounds may also function as inhibitors of inosine- 5’ -monophosphate dehydrogenase (IMPDH). Further, in certain embodiments, the invention provides pharmaceutical compositions comprising one or more compounds described herein, and a pharmaceutically acceptable excipient. In certain embodiments, the invention also relates to methods of treating various bacterial infections in mammals. Moreover, the compounds may be used alone or in combination with other therapeutic or prophylactic agents, such as antibacterial agents.

In certain embodiments, the invention relates to compounds that induce bacteria to initiate growth prematurely (Figure 3A). In certain embodiments, IMPDH inhibitors cause quiescent cells to become sensitive to commonly used antibiotics, thus providing a more effective treatment for chronic infections. In certain embodiments, the compounds increase the bacteriocidal activity of commonly used antibiotics, as expected if persister cells are induced to grow (Figure 3B).

In certain embodiments, the compositions and methods described herein are effective in the presence of a biofilm.

Exemplary Compounds

In certain embodiments, the invention relates to a compound having the following structure:

, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a compound having a structure selected from the group consisting of:

Exemplary Pharmaceutical Compositions

In certain embodiments, the invention relates to a pharmaceutical composition, comprising a pharmaceutically acceptable carrier, adjuvant, or vehicle; and a compound having the following structure:

pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a pharmaceutical composition, comprising

(a) a compound having the following structure:

, or a pharmaceutically acceptable salt thereof;

(b) an antibacterial agent; and

(c) a pharmaceutically acceptable carrier, adjuvant, or vehicle.

In certain embodiments, the invention relates to a pharmaceutical composition, comprising a pharmaceutically acceptable carrier, adjuvant, or vehicle; and a compound having a structure selected from the group consisting of:

pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a pharmaceutical composition, comprising

(a) a compound having a structure selected from the group consisting of:

pharmaceutically acceptable salt thereof; (b) an antibacterial agent; and

(c) a pharmaceutically acceptable carrier, adjuvant, or vehicle.

In certain embodiments, the invention relates to a pharmaceutical composition, comprising

(a) any one of the compounds described in US Patent Application Publication No. US 2010/0022547, US Patent No. 8,969,342, or US Patent No. 9,447,134, each of which is incorporated by reference in its entirety;

(b) an antibacterial agent; and

(c) a pharmaceutically acceptable carrier, adjuvant, or vehicle.

In certain embodiments, the invention relates to a pharmaceutical composition, comprising

(a) a compound selected from the group consisting of:

(b) an antibacterial agent; and

(c) a pharmaceutically acceptable carrier, adjuvant, or vehicle.

In certain embodiments, the invention relates to a pharmaceutical composition, comprising

(a) a compound selected from the group consisting of:

(b) an antibacterial agent; and

(c) a pharmaceutically acceptable carrier, adjuvant, or vehicle.

In certain embodiments, the invention relates to a pharmaceutical composition, comprising

(a) a compound having the following structure:

(b) an antibacterial agent; and

(c) a pharmaceutically acceptable carrier, adjuvant, or vehicle.

In certain embodiments, the antibacterial agent is an anti-translation agent, for example, an agent that interferes with bacterial protein synthesis, for example, by interacting with prokaryotic ribosomes. In certain embodiments, the antibacterial agent inhibits DNA gyrase or a topoisomerase in a bacterium. In certain embodiments, the antibacterial agent interferes with the transpeptidation reaction, thereby inhibits synthesis of peptidoglycan, a component of bacterial cell walls.

In certain embodiments, the antibacterial agent is a penicillin. In certain embodiments, the antibacterial agent is a tetracycline. In certain embodiments, the antibacterial agent is a cephalosporin. In certain embodiments, the antibacterial agent is a quinolone, such as a fluoroquinolone. In certain embodiments, the antibacterial agent is a linomycin. In certain embodiments, the antibacterial agent is a macrolide. In certain embodiments, the antibacterial agent is a sulfonamide. In certain embodiments, the antibacterial agent is a glycopeptide. In certain embodiments, the antibacterial agent is an aminoglycoside. In certain embodiments, the antibacterial agent is a carbapenem.

In certain embodiments, the invention relates to any one of the compositions described herein, further comprising an antibacterial agent selected from the group consisting of amikacin, amoxicillin, azithromycin, cefdinir, ceftriaxone, cefuroxime, cephalexin, ciprofloxacin, clarithromycin, clindamycin, co-trimoxazole, dalbavancin, doripenem, doxycycline, ertapenem, erythromycin, gentamicin, imipenem/cilastatin, kanamycin, levofloxacin, lincomycin, meropenem, metronidazole, minocycline, moxifloxacin, ofloxacin, oritavancin, oxacillin, penicillin, sulfamethoxazole, sulfasalazine, sulfisoxazole, telavancin, tetracycline, tobramycin, trimethoprim, and vancomycin.

The pharmaceutical compositions of the invention may be administered orally, parenternally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The pharmaceutical compositions of the invention may contain any conventional non-toxic pharmaceutically-acceptable carriers, adjuvants or vehicles. In some cases, the pH of the formulation may be adjusted with pharmaceutically acceptable acids, bases or buffers to enhance the stability of the formulated compound or its delivery form. The term parentemal as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intra-articular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.

The pharmaceutical compositions may be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are mannitol, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant such as those described in Pharmacopeia Helvetica, Ph. Helv., or a similar alcohol, or carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms such as emulsions and or suspensions. Other commonly used surfactants such as Tweens or Spans and/or other similar emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.

The pharmaceutical compositions of the invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, emulsions and aqueous suspensions, dispersions and solutions. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried com starch. When aqueous suspensions and/or emulsions are administered orally, the active ingredient may be suspended or dissolved in an oily phase is combined with emulsifying and/or suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added.

The pharmaceutical compositions of the invention may also be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing a compound of the invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components. Such materials include, but are not limited to, cocoa butter, beeswax and polyethylene glycols.

Topical administration of the pharmaceutical compositions of the invention is especially useful when the desired treatment involves areas or organs readily accessible by topical application. For application topically to the skin, the pharmaceutical composition should be formulated with a suitable ointment containing the active components suspended or dissolved in a carrier. Carriers for topical administration of the compounds of the invention include, but are not limited to, mineral oil, liquid petroleum, white petroleum, propylene glycol, polyoxy-ethylene polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical composition can be formulated with a suitable lotion or cream containing the active compound suspended or dissolved in a carrier with suitable emulsifying agents. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. The pharmaceutical compositions of the invention may also be topically applied to the lower intestinal tract by rectal suppository formulation or in a suitable enema formulation. Topically-transdermal patches are also included in this invention.

The pharmaceutical compositions of the invention may be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art.

Dosage levels of between about 0.01 and about 1000 mg/kg body weight per day, or between about 0.5 and about 75 mg/kg body weight per day, of the IMPDH inhibitory compounds described herein are useful in a monotherapy and/or in combination therapy for the prevention and treatment of IMPDH-mediated disease or infection. Typically, the pharmaceutical compositions of the invention will be administered from about 1 to about 5 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. A typical preparation will contain from about 5% to about 95% active compound (w/w). Such preparations contain from about 20% to about 80% active compound.

When the compositions of the invention comprise a combination of an antibacterial agent and a compound described herein, or when the compositions of the invention comprise a combination of an antibacterial agent and a compound described in US 2010/0022547, US 8,969,342, or US 9,447, 134, both agents should be present at dosage levels of between about 10 to 100%, or between about 10 to 80% of the dosage normally administered in a monotherapy regimen. Upon improvement of a patient's condition, a maintenance dose of a compound, composition or combination of the invention may be administered, if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained when the symptoms have been alleviated to the desired level, treatment should cease. Patients may, however, require intermittent treatment on a long-term basis upon any recurrence of disease symptoms.

As the skilled artisan will appreciate, lower or higher doses than those recited above may be required. Specific dosage and treatment regimens for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health status, sex, diet, time of administration, rate of excretion, drug combination, the severity and course of the infection, the patient's disposition to the infection and the judgment of the treating physician.

Combination Therapy

One aspect of the present invention relates to combination therapy. This type of therapy is advantageous because the co-administration of active ingredients achieves a therapeutic effect that is greater than the therapeutic effect achieved by administration of only a single therapeutic agent.

In certain embodiments, the co-administration of two or more therapeutic agents achieves a therapeutic effect that is greater than the therapeutic effect achieved by administration of only a single therapeutic agent. In this regard, the combination therapies are efficacious. The therapeutic effect of one therapeutic agent is augmented by the co administration of another therapeutic agent.

In certain embodiments, the co-administration of two or more therapeutic agents achieves a therapeutic effect that is equal to about the sum of the therapeutic effects achieved by administration of each single therapeutic agent. In these embodiments, the combination therapies are said to be“additive.”

In certain embodiments, the co-administration of two or more therapeutic agents achieves a synergistic effect, i.e., a therapeutic effect that is greater than the sum of the therapeutic effects of the individual components of the combination.

The active ingredients that comprise a combination therapy may be administered together via a single dosage form or by separate administration of each active agent. In certain embodiments, the first and second therapeutic agents are administered in a single dosage form. In certain embodiments, the first, second, and third therapeutic agents are administered in a single dosage form. The agents may be formulated into a single tablet, pill, capsule, or solution for parenteral administration and the like.

In certain embodiments, the therapeutic agents are administered in a single dosage form, wherein each individual therapeutic agent is isolated from the other therapeutic agent(s). Formulating the dosage forms in such a way assists in maintaining the structural integrity of potentially reactive therapeutic agents until they are administered. A formulation of this type may be useful during production and for long-term storage of the dosage form. In certain embodiments, the therapeutic agents may comprise segregated regions or distinct caplets or the like housed within a capsule. In certain embodiments, the therapeutic agents are provided in isolated layers comprised by a tablet.

Alternatively, the therapeutic agents may be administered as separate compositions, e.g., as separate tablets or solutions. One or more active agent may be administered at the same time as the other active agent(s) or the active agents may be administered intermittently. The length of time between administrations of the therapeutic agents may be adjusted to achieve the desired therapeutic effect. In certain instances, one or more therapeutic agent(s) may be administered only a few minutes (e.g., about 1, 2, 5, 10, 30, or 60 min) after administration of the other therapeutic agent(s). Alternatively, one or more therapeutic agent(s) may be administered several hours (e.g., about 2, 4, 6, 10, 12, 24, or 36 h) after administration of the other therapeutic agent(s). In certain embodiments, it may be advantageous to administer more than one dosage of one or more therapeutic agent(s) between administrations of the remaining therapeutic agent(s). For example, one therapeutic agent may be administered at 2 hours and then again at 10 hours following administration of the other therapeutic agent(s). Importantly, it is required that the therapeutic effects of each active ingredient overlap for at least a portion of the duration of each therapeutic agent so that the overall therapeutic effect of the combination therapy is attributable in part to the combined or synergistic effects of the combination therapy.

The dosage of the active agents will generally be dependent upon a number of factors including pharmacodynamic characteristics of each agent of the combination, mode and route of administration of active agent(s), the health of the patient being treated, the extent of treatment desired, the nature and kind of concurrent therapy, if any, and the frequency of treatment and the nature of the effect desired. In general, dosage ranges of the active agents often range from about 0.001 to about 250 mg/kg body weight per day. For a normal adult having a body weight of about 70 kg, a dosage in the range of from about 0.1 to about 25 mg/kg body weight is typically preferred. However, some variability in this general dosage range may be required depending upon the age and weight of the subj ect being treated, the intended route of administration, the particular agent being administered and the like. Since two or more different active agents are being used together in a combination therapy, the potency of each agent and the interactive effects achieved using them together must be considered. Importantly, the determination of dosage ranges and optimal dosages for a particular mammal is also well within the ability of one of ordinary skill in the art having the benefit of the instant disclosure.

In certain embodiments, it may be advantageous for the pharmaceutical combination to have a relatively large amount of the first component compared to the second component. In certain instances, the ratio of the first active agent to second active agent is about 100: 1, 90: 1, 80: 1, 70: 1, 60: 1, 50: 1, 40: 1, 30: 1, 20: 1, 15: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, or 5: 1. In certain embodiments, it may be preferable to have a more equal distribution of pharmaceutical agents. In certain instances, the ratio of the first active agent to the second active agent is about 4: 1, 3 : 1, 2: 1, 1 : 1, 1 :2, 1 :3, or 1 :4. In certain embodiments, it may be advantageous for the pharmaceutical combination to have a relatively large amount of the second component compared to the first component. In certain instances, the ratio of the second active agent to the first active agent is about 30: 1, 20: 1, 15: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, or 5: 1. In certain instances, the ratio of the second active agent to first active agent is about 100: 1, 90: 1, 80: 1, 70: 1, 60: 1, 50: 1, or 40: 1. Importantly, a composition comprising any of the above-identified combinations of first therapeutic agent and second therapeutic agent may be administered in divided doses about 1, 2, 3, 4, 5, 6, or more times per day or in a form that will provide a rate of release effective to attain the desired results. In one embodiment, the dosage form contains both the first and second active agents. In one embodiment, the dosage form only has to be administered one time per day and the dosage form contains both the first and second active agents.

For example, a formulation intended for intravenous administration to humans may contain from about 0.1 mg to about 5 g of the first therapeutic agent and about 0.1 mg to about 5 g of the second therapeutic agent, both of which are compounded with an appropriate and convenient amount of carrier material varying from about 5 to about 95 percent of the total composition. Unit dosages will generally contain between about 0.5 mg to about 1500 mg of the first therapeutic agent and 0.5 mg to about 1500 mg of the second therapeutic agent. In a preferred embodiment, the dosage is about 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg, etc., up to about 1500 mg of the first therapeutic agent. In a preferred embodiment, the dosage is about 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg, etc., up to about 1500 mg of the second therapeutic agent.

Dosage amount and interval may be adjusted on an individual or group basis to provide plasma levels of a particular active moiety or moieties sufficient to maintain the modulating effects or minimal effective concentration (MEC) of each of them. The MEC will vary for each compound and individual, but it can be estimated from in vitro data. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. However, HPLC assays or bioassays can be used to determine plasma concentrations. In certain embodiments, the dose may be decreased. In certain embodiments, the dose may be increased. Moreover, a long-term treatment regimen may include alternating period of increasing and decreasing dosage with respect to a particular compound or compounds.

Synergism and Augmentation

The term "synergistic" refers to a combination which is more effective than the additive effects of any two or more single agents. A synergistic effect permits the effective treatment of a disease using lower amounts (doses) of individual therapy. The lower doses result in lower toxicity without reduced efficacy. In addition, a synergistic effect can result in improved efficacy. Finally, synergy may result in an improved avoidance or reduction of disease as compared to any single therapy.

Combination therapy can allow for the product of lower doses of the first therapeutic or the second therapeutic agent (referred to as "apparent one-way synergy" herein), or lower doses of both therapeutic agents (referred to as "two-way synergy" herein) than would normally be required when either drug is used alone.

Combination therapy can allow for the product of lower doses of any one of the therapeutic agents (referred to as "apparent one-way synergy" herein), or lower doses of all therapeutic agents than would normally be required when any drug is used alone.

In certain embodiments, the synergism exhibited between one or more therapeutic agent(s) and the remaining therapeutic agent(s) is such that the dosage of one of the therapeutic agents would be sub-therapeutic if administered without the dosage of the other therapeutic agents. The terms“augmentation” or“augment” refer to combinations where one of the compounds increases or enhances therapeutic effects of another compound or compounds administered to a patient. In some instances, augmentation can result in improving the efficacy, tolerability, or safety, or any combination thereof, of a particular therapy.

In certain embodiments, the present invention relates to a pharmaceutical composition comprising a therapeutically effective dose of one or more therapeutic agent(s) together with a dose of another therapeutic agent effective to augment the therapeutic effect of the one or more therapeutic agent(s). In other embodiments, the present invention relates to methods of augmenting the therapeutic effect in a patient of one or more therapeutic agent(s) by administering another therapeutic agent to the patient.

In certain preferred embodiments, the invention is directed in part to synergistic combinations of one or more therapeutic agent(s) in an amount sufficient to render a therapeutic effect together with the remaining therapeutic agent(s). For example, in certain embodiments a therapeutic effect is attained which is at least about 2 (or at least about 4, 6, 8, or 10) times greater than that obtained with the dose of the one or more therapeutic agent(s) alone. In certain embodiments, the synergistic combination provides a therapeutic effect which is up to about 20, 30 or 40 times greater than that obtained with the dose of the one or more therapeutic agent(s) alone. In such embodiments, the synergistic combinations display what is referred to herein as an "apparent one-way synergy", meaning that the dose of the remaining therapeutic agent(s) synergistically potentiates the effect of the one or more therapeutic agent(s), but the dose of the one or more therapeutic agent(s) does not appear to significantly potentiate the effect of the remaining therapeutic agent(s).

In certain embodiments, the combination of active agents exhibits two-way synergism, meaning that the second therapeutic agent potentiates the effect of the first therapeutic agent, and the first therapeutic agent potentiates the effect of the second therapeutic agent. Thus, other embodiments of the invention relate to combinations of a second therapeutic agent and a first therapeutic agent where the dose of each drug is reduced due to the synergism between the drugs, and the therapeutic effect derived from the combination of drugs in reduced doses is enhanced. The two-way synergism is not always readily apparent in actual dosages due to the potency ratio of the first therapeutic agent to the second therapeutic agent. For instance, two-way synergism can be difficult to detect when one therapeutic agent displays much greater therapeutic potency relative to the other therapeutic agent. The synergistic effects of combination therapy may be evaluated by biological activity assays. For example, the therapeutic agents are mixed at molar ratios designed to give approximately equipotent therapeutic effects based on the EC90 values. Then, three different molar ratios are used for each combination to allow for variability in the estimates of relative potency. These molar ratios are maintained throughout the dilution series. The corresponding monotherapies are also evaluated in parallel to the combination treatments using the standard primary assay format. A comparison of the therapeutic effect of the combination treatment to the therapeutic effect of the monotherapy gives a measure of the synergistic effect. Further details on the design of combination analyses can be found in B E Korba (1996) Antiviral Res. 29:49. Analysis of synergism, additivity, or antagonism can be determined by analysis of the aforementioned data using the CalcuSyn™ program (Biosoft, Inc.). This program evaluates drug interactions by use of the widely accepted method of Chou and Talalay combined with a statistically evaluation using the Monte Carlo statistical package. The data are displayed in several different formats including median-effect and dose-effects plots, isobolograms, and combination index [Cl] plots with standard deviations. For the latter analysis, a Cl greater than 1.0 indicates antagonism and a Cl less than 1.0 indicates synergism.

Compositions of the invention present the opportunity for obtaining relief from moderate to severe cases of disease. Due to the synergistic or additive or augmented effects provided by the inventive combination of the first and second therapeutic agent, it may be possible to use reduced dosages of each of therapeutic agent. Due to the synergistic or additive or augmented effects provided by the inventive combination of the first, second, and third therapeutic agents, it may be possible to use reduced dosages of each of therapeutic agent. By using lesser amounts of drugs, the side effects associated with each may be reduced in number and degree. Moreover, the inventive combinations avoid side effects to which some patients are particularly sensitive.

Exemplary Methods

In certain embodiments, the invention relates to a method of killing or inhibiting the growth of a microbe, comprising the step of contacting said microbe with an effective amount of a compound having the following structure: , or a pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a method of killing or inhibiting the growth of a microbe, comprising the step of contacting said microbe with an effective amount of a compound having a structure selected from the group consisting of:

In certain embodiments, the invention relates to any one of the methods described herein, wherein said microbe is a protozoan or bacterium.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said microbe is a protozoan or a bacterium selected from the group consisting of the genera Acinetobacter, Arcobacter , Bacillus, Bacteroides, Borrelia, Brucella , Burkholderia, Campylobacter, Clostridia , Coxiella , Cryptosporidium, Entamoeba, Enterococcus , Erysipelothrix , Francisella , Fusobacterium, Helicobacter , Lactobacillus , Leishmania , Listeria , Mycobacterium, Neisseria , Pseudomonas , Staphylococcus , Streptococcus , and Trypanosoma.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said microbe is a protozoan or a bacterium selected from the group consisting of Acinetobacter baumannii , Bacillus anthracis , Burkholderia cenocepacia , thailandensis , Campylobacter jejuni , Francisella tularensis , Helicobacter pylori , Staphylococcus aureus , Listeria monocytogenes, and Mycobacterium tuberculosis.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said microbe is a protozoan or a bacterium selected from the group consisting of Acinetobacter baumannii ATCC 17961, Bacillus anthracis Sterne 7702, Burkholderia cenocepacia K56-2, B. thailandensis E264, Campylobacter jejuni 81-176, Francisella tularensis Schu S4, Helicobacter pylori , Staphylococcus aureus NCTC 8325, Listeria monocytogenes 10403S, and Mycobacterium tuberculosis H 37 Rv .

In certain embodiments, the invention relates to any one of the methods described herein, wherein said microbe is a protozoan; and said protozoan is selected from the group consisting of the genera Cryptosporidium, Entamoeba, Leishmania and Trypanosoma.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said protozoan is selected from the genus Cryptosporidium.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said protozoan is Cryptosporidium parvum and/or Cryptosporidium hominis.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said microbe is a bacterium; and said bacterium is selected from the group consisting of the genera Acinetobacter, Arcobacter , Bacillus, Bacteroides, Borrelia, Brucella , Burkholderia, Brachyspira, Campylobacter, Clostridia , Coxiella , Enterococcus , Erysipelothrix , Francisella , Fusobacterium, Helicobacter , Lactobacillus , Listeria , Mycobacterium, Neisseria , Pseudomonas , Staphylococcus and Streptococcus.

In certain embodiments, the invention relates to a method of treating or preventing a microbial infection in a mammal, comprising the step of administering to a mammal in need thereof a therapeutically effective amount of a compound having the following structure:

, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a method of treating or preventing a microbial infection in a mammal, comprising the step of administering to a mammal in need thereof a therapeutically effective amount of a compound having a structure selected from the group consisting of:

In certain embodiments, the invention relates to any one of the methods described herein, wherein the method is a method of treating a microbial infection in a mammal.

In certain embodiments, the invention relates to a method of treating or preventing a parasitic infection in a mammal comprising the step of administering to a mammal in need thereof a therapeutically effective amount of a compound having the following structure:

, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the invention relates to a method of treating or preventing a parasitic infection in a mammal comprising the step of administering to a mammal in need thereof a therapeutically effective amount of a compound having a structure selected from the group consisting of:

In certain embodiments, the invention relates to any one of the methods described herein, wherein the method is a method of treating a parasitic infection in a mammal.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said microbial infection is caused by a protozoan or bacterium.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said microbial infection is caused by a protozoan or a bacterium selected from the group consisting of the genera Cryptosporidium, Entamoeba, Leishmania , Trypanosoma, Acinetobacter, Arcobacter , Bacillus, Bacteroides, Borrelia, Brucella , Burkholderia, Brachyspira, Campylobacter, Clostridia , Coxiella , Enterococcus , Erysipelothrix , Francisella , Fusobacterium, Helicobacter , Lactobacillus , Listeria , Mycobacterium, Neisseria , Pseudomonas , Staphylococcus and Streptococcus.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said microbial infection is caused by a protozoan; and said protozoan is selected from the group consisting of the genera Cryptosporidium, Entamoeba, Leishmania and Trypanosoma.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said protozoan is selected from the genus Cryptosporidium. In certain embodiments, the invention relates to any one of the methods described herein, wherein said microbial infection is caused by Cryptosporidium parvum or Cryptosporidium hominis or both.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said microbe is a bacterium; and said bacterium is selected from the group consisting of the genera Acinetobacter, Arcobacter , Bacillus, Bacteroides, Borrelia, Brucella , Burkholderia, Brachyspira, Campylobacter, Clostridia , Coxiella , Enterococcus , Erysipelothrix , Francisella , Fusobacterium, Helicobacter , Lactobacillus , Listeria , Mycobacterium, Neisseria , Pseudomonas , Staphylococcus and Streptococcus.

In certain embodiments, the invention relates to a method of treating or preventing a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of a compound having the following structure:

, or a pharmaceutically acceptable salt thereof; and

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are effective.

In certain embodiments, the invention relates to a method of treating or preventing a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of a compound having a structure selected from the group consisting

and

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are effective.

In certain embodiments, the invention relates to any one of the methods described herein, wherein the method is a method of treating a bacterial infection in a mammal; and, taken together, the first amount and the second amount are therapeutically effective.

In certain embodiments, the invention relates to a method of treating a recurrent or persistent bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of any one of the compounds described in US Patent Application Publication No. US 2010/0022547, US Patent No. 8,969,342, or US Patent No. 9,447,134, each of which is incorporated by reference in its entirety; and

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are therapeutically effective. In certain embodiments, the invention relates to a method of treating a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of any one of the compounds described in US Patent Application Publication No. US 2010/0022547, US Patent No. 8,969,342, or US Patent No. 9,447,134, each of which is incorporated by reference in its entirety; and

(b) a second amount of an antibacterial agent,

wherein,

taken together, the first amount and the second amount are therapeutically effective; and

the bacterial infection does not persist or recur after treatment.

In certain embodiments, the invention relates to a method of preventing the recurrence of a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of any one of the compounds described in US Patent Application Publication No. US 2010/0022547, US Patent No. 8,969,342, or US Patent No. 9,447,134, each of which is incorporated by reference in its entirety; and

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are effective to prevent the recurrence of the bacterial infection in the mammal.

In certain embodiments, the invention relates to a method of treating a recurrent or persistent bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of a compound selected from the group consisting of:

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are therapeutically effective. In certain embodiments, the invention relates to a method of treating a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof (a) a first amount of a compound selected from the group consisting of:

(b) a second amount of an antibacterial agent,

wherein,

taken together, the first amount and the second amount are therapeutically effective; and

the bacterial infection does not persist or recur after treatment.

In certain embodiments, the invention relates to a method of preventing the recurrence of a bacterial infection in a mammal, comprising the step of co-administering to a mammal in need thereof

(a) a first amount of a compound selected from the group consisting of:

(b) a second amount of an antibacterial agent,

wherein, taken together, the first amount and the second amount are effective to prevent the recurrence of the bacterial infection in the mammal.

In certain embodiments, the invention relates to any one of the methods described herein, wherein (a) and (b) are co-administered in the form of any one of the pharmaceutical compositions described herein.

In certain embodiments, the invention relates to any one of the methods described herein, wherein said bacterium is selected from the group consisting of the genera Acinetobacter, Arcobacter , Bacillus, Bacteroides, Borrelia, Brucella , Burkholderia, Brachyspira, Campylobacter, Clostridia , Coxiella , Enterococcus , Erysipelothrix , Francisella , Fusobacterium, Helicobacter , Lactobacillus , Listeria , Mycobacterium, Neisseria , Pseudomonas , Staphylococcus and Streptococcus.

In certain embodiments, the antibacterial agent is an anti -translation agent, for example, an agent that interferes with bacterial protein synthesis, for example, by interacting with prokaryotic ribosomes. In certain embodiments, the antibacterial agent inhibits DNA gyrase or a topoisomerase in a bacterium. In certain embodiments, the antibacterial agent interferes with the transpeptidation reaction, thereby inhibits synthesis of peptidoglycan, a component of bacterial cell walls. In certain embodiments, the antibacterial agent is a penicillin. In certain embodiments, the antibacterial agent is a tetracycline. In certain embodiments, the antibacterial agent is a cephalosporin. In certain embodiments, the antibacterial agent is a quinolone, such as a fluoroquinolone. In certain embodiments, the antibacterial agent is a linomycin. In certain embodiments, the antibacterial agent is a macrolide. In certain embodiments, the antibacterial agent is a sulfonamide. In certain embodiments, the antibacterial agent is a glycopeptide. In certain embodiments, the antibacterial agent is an aminoglycoside. In certain embodiments, the antibacterial agent is a carbapenem.

In certain embodiments, the invention relates to any one of the methods described herein, further comprising an antibacterial agent selected from the group consisting of amikacin, amoxicillin, azithromycin, cefdinir, ceftriaxone, cefuroxime, cephalexin, ciprofloxacin, clarithromycin, clindamycin, co-trimoxazole, dalbavancin, doripenem, doxycycline, ertapenem, erythromycin, gentamicin, imipenem/cilastatin, kanamycin, levofloxacin, lincomycin, meropenem, metronidazole, minocycline, moxifloxacin, ofloxacin, oritavancin, oxacillin, penicillin, sulfamethoxazole, sulfasalazine, sulfisoxazole, telavancin, tetracycline, tobramycin, trimethoprim, and vancomycin.

In certain embodiments, the invention relates to any one of the methods described herein, wherein, prior to co-administration of (a) and (b), the mammal had been administered a first antibacterial agent. In certain embodiments, the first antibacterial agent is selected from the group consisting of amikacin, amoxicillin, azithromycin, cefdinir, ceftriaxone, cefuroxime, cephalexin, ciprofloxacin, clarithromycin, clindamycin, co-trimoxazole, dalbavancin, doripenem, doxycycline, ertapenem, erythromycin, gentamicin, imipenem/cilastatin, kanamycin, levofloxacin, lincomycin, meropenem, metronidazole, minocycline, moxifloxacin, ofloxacin, oritavancin, oxacillin, penicillin, sulfamethoxazole, sulfasalazine, sulfisoxazole, telavancin, tetracycline, tobramycin, trimethoprim, and vancomycin. In certain embodiments, the first antibacterial agent and the antibacterial agent are the same. In certain embodiments, the first antibacterial agent and the antibacterial agent are not the same.

Definitions

All definitions, as defined and used herein, should be understood to control over dictionary definitions, definitions in documents incorporated by reference, and/or ordinary meanings of the defined terms. The indefinite articles“a” and“an,” as used herein in the specification and in the claims, unless clearly indicated to the contrary, should be understood to mean“at least one.”

The phrase“and/or,” as used herein in the specification and in the claims, should be understood to mean“either or both” of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with“and/or” should be construed in the same fashion, i.e.,“one or more” of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the“and/or” clause, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, a reference to“A and/or B”, when used in conjunction with open-ended language such as“comprising” can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.

As used herein in the specification and in the claims,“or” should be understood to have the same meaning as“and/or” as defined above. For example, when separating items in a list,“or” or“and/or” shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as“only one of’ or“exactly one of,” or, when used in the claims,“consisting of,” will refer to the inclusion of exactly one element of a number or list of elements. In general, the term“or” as used herein shall only be interpreted as indicating exclusive alternatives (i.e.,“one or the other but not both”) when preceded by terms of exclusivity, such as“either,”“one of,”“only one of,” or “exactly one of.”“Consisting essentially of,” when used in the claims, shall have its ordinary meaning as used in the field of patent law.

As used herein in the specification and in the claims, the phrase“at least one,” in reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase“at least one” refers, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example,“at least one of A and B” (or, equivalently,“at least one of A or B,” or, equivalently“at least one of A and/or B”) can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.

It should also be understood that, unless clearly indicated to the contrary, in any methods claimed herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited.

In the claims, as well as in the specification above, all transitional phrases such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” “composed of,” and the like are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases“consisting of’ and“consisting essentially of’ shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03.

Certain compounds contained in compositions of the present invention may exist in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, R- and //-enantiomers, diastereomers, (D)- i somers, (L)-i somers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention. All such isomers, as well as mixtures thereof, are intended to be included in this invention.

If, for instance, a particular enantiomer of compound of the present invention is desired, it may be prepared by asymmetric synthesis, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers. Alternatively, where the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means well known in the art, and subsequent recovery of the pure enantiomers. For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, “Handbook of Chemistry and Physics”, 67th Ed., 1986-87, inside cover.

A“pharmaceutically acceptable salt” means any pharmaceutically acceptable salt, which, upon administration to a recipient, is capable of providing (directly or indirectly) a compound of this invention.

Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acid salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2- hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, nitrate, oxalate, palmoate, pectinate, persulfate, 3- phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, succinate, sulfate, tartrate, thiocyanate, tosylate and undecanoate. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts.

Salts derived from appropriate bases include alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), and ammonium salts. This invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil-soluble or dispersible products may be obtained by such quaternization.

The term“pharmaceutically acceptable carrier or adjuvant” refers to a carrier or adjuvant that may be administered to a patient, together with a therapeutic agent, and which does not destroy the pharmacological activity thereof and is nontoxic when administered in doses sufficient to deliver a therapeutic amount of the agent.

Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions of the invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) such as d. alpha. -tocopherol poly ethyleneglycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens or other similar polymeric delivery matrices, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat. Cyclodextrins such as a-, b-, and g-cyclodextrin, or chemically modified derivatives such as hydroxyalkylcyclodextrins, including 2- and 3- hydroxypropyl-P-cyclodextrins, or other solubilized derivatives may also be advantageously used to enhance delivery of any one of the agents.

The term“treating” as used herein refers to the alleviation of symptoms of a particular disorder in a patient or the improvement of an ascertainable measurement associated with a particular disorder. As used herein, the term“patient” refers to a mammal, including a human.

The terms “co-administration” and “co-administering” refer to both concurrent administration (administration of two or more therapeutic agents at the same time) and time varied administration (administration of one or more therapeutic agents at a time different from that of the administration of an additional therapeutic agent or agents), as long as the therapeutic agents are present in the patient to some extent at the same time.

A “recurrent” or “persistent” bacterial infection results from certain bacterial pathogens that are able to evade the host immune system and persist within the human host. The consequences of persistent bacterial infections potentially include increased morbidity and mortality from the infection itself as well as an increased risk of dissemination of disease. Eradication of persistent infections is difficult, often requiring prolonged or repeated courses of antibiotics. During persistent infections, a population or subpopulation of bacteria exists that is refractory to traditional antibiotics, possibly in a non-replicating or metabolically altered (“quiescent”) state. As used herein,“preventing the recurrence of’ an infection describes eradication of the agent that caused the original infection. The efficacy of preventing the recurrence of the infection is determined by a successful clinical outcome and does not require 100% elimination of the microorganisms involved in the infection. Achieving a level of antimicrobial activity at the site of infection that allows the host to survive, resolve the infection, or eradicate the causative agent is sufficient.

While several embodiments of the present invention are described and illustrated herein, those of ordinary skill in the art will readily envision a variety of other means and/or structures for performing the functions and/or obtaining the results and/or one or more of the advantages described herein, and each of such variations and/or modifications is deemed to be within the scope of the present invention. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and/or configurations will depend upon the specific application or applications for which the teachings of the present invention is/are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and equivalents thereto, the invention may be practiced otherwise than as specifically described and claimed. The present invention is directed to each individual feature, system, article, material, kit, and/or method described herein. In addition, any combination of two or more such features, systems, articles, materials, kits, and/or methods, if such features, systems, articles, materials, kits, and/or methods are not mutually inconsistent, is included within the scope of the present invention.

EXEMPLIFICATION

The invention now being generally described, it will be more readily understood by reference to the following examples which are included merely for purposes of illustration of certain aspects and embodiments of the present invention, and are not intended to limit the invention.

Example 1 - P226 sensitizes bacteria to conventional antibiotics

P226 sensitizes Staph bacteria to the conventional antibiotics rifampicin, ciprofloxacin and oxacillin. See Figure 2 and Figures 3A-3C.

Prophetic Example 2 - Determine if P226 and related compounds sensitize persister cells in biofilms to conventional antibiotics

The effects of P226 and related compounds on S. aureus biofilms in combination with conventional antibiotics such as rifampicin, ciprofloxacin, vancomycin and oxacillin is investigated. Prophetic Example 3 - Determine efficacy of P226 or a derivative in a mouse model of abscess infection

P226 is applied directly into an abscess, thus avoiding potential issues with bioavailability. The ability of P226 to sterilize abscesses in combination with a conventional antibiotic is investigated.

INCORPORATION BY REFERENCE

All of the U.S. patents and U.S. published patent applications cited herein are hereby incorporated by reference.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.