Compositions and synergistic methods for treating infections

Abstract

The present invention relates to compositions and methods for treating a microbial infection in a subject, particularly methods of administering a gelsolin agent and an antibacterial agent to produce a synergistic therapeutic effect against a microbial infection in a subject. The present invention also relates to methods for treating a viral infection in a subject, including methods involving delayed dosing and / or synergistic methods.

Description

Technical Field

[0001] Related Applications This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application No. 62 / 864,599, filed Jun. 21, 2019, the disclosure of which is hereby incorporated by reference in its entirety. Government Interests This invention was made with government support under grant NIH AI125152 and NIH / NIAID contracts HHSN272201000033I-HHSN27200003 and HHSN272201000033I-HHSN27200006. The U.S. government has certain rights in this invention.

[0002] In some aspects, the present invention relates to compositions and methods for enhancing host immune defenses in the treatment of microbial infections.

Background Art

[0003] Antimicrobial resistance is a global public health concern. Antimicrobial resistance is known to reduce the therapeutic efficacy of various antimicrobial agents such as antibacterial agents, antiviral agents, antifungal agents, and antiparasitic agents. Examples of the evolving presence of resistant pneumococcal species include case reports of fatal resistant pneumococcal pneumonia (Waterer GW et al., Chest 2000;18:1839-1840) and the finding that 22% (139 / 643) of patients admitted for S. pneumoniae had macrolide-resistant bacteria (Cilloniz et al., Am J Respir Crit Care Med 2015;191:1265-1272). Recent publications have demonstrated (1) resistance of S. pneumoniae isolates from invasive infections to erythromycin (96%), trimethoprim-sulfamethoxazole (79%) and tetracycline (77%) in the pediatric population (Cai et al., Infect Drug Resist 2018;11:2461-2469) and (2) investigation of S. pneumoniae isolates from invasive infections in the elderly population (Intra et al., Front Public Health 2017;5:169). The review publication, Kollef & Betthauser, Curr.Opin.Inf.Dis.2019;32:169-175 highlights the increasing antibiotic resistance of common pathogens associated with community-acquired pneumonia (CAP), particularly staphylococci and Streptococcus pneumoniae.

[0004] Antimicrobial agents have been used for a long time to treat microbial infections because they have a therapeutic effect against microbial infections in humans and animals. Resistance to antimicrobial agents that were previously therapeutically effective may be caused by changes in the pathogens that cause the infection. The misuse and overuse of antibacterial drugs can be factors contributing to the spread of the problem of antimicrobial resistance, which has led to an increasing number of types of pathogenic infections with reduced responsiveness to antibacterial agents that were previously effective. Antimicrobial resistance results in a lack of treatment options for treating pathogenic infections. Antimicrobial-resistant pathogens cause many deaths every year and are a serious global public health problem.

Summary of the Invention

Means for Solving the Problems

[0005] The present invention relates, in part, to a composition that can be used to synergistically treat microbial infections. The composition includes one or more antimicrobial agents and a gelsecrin agent. The method of the present invention relates, in part, to the administration of such a composition to a subject, and the antimicrobial agent and the gelsecrin agent act synergistically to treat a microbial infection in the subject.

[0006] According to one aspect of the present invention, there is provided a composition comprising an effective amount of a gelseolin agent and an antibacterial agent for synergistically treating microbial infections in a subject. In some embodiments, the antibacterial agent is in a clinically acceptable amount, and the administered gelseolin agent and antibacterial agent synergistically enhance the therapeutic effect of administering a clinically acceptable amount of the antibacterial agent to the subject without administering the gelseolin agent. In certain embodiments, the clinically acceptable amount of the antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent in the subject. In some embodiments, the MTD of the antibacterial agent is the highest possible but still acceptable dosage level of the antibacterial agent for the subject. In some embodiments, the MTD of the antibacterial agent is determined based at least in part on a preselected clinical limiting toxicity for the antibacterial agent in the subject. In certain embodiments, the synergistic effective amount of the gelseolin agent and the antibacterial agent reduces the minimum effective dose (MED) of the antibacterial agent in the subject. In certain embodiments, the MED is the lowest dosage level of the antibacterial agent that provides a clinically significant response in mean effectiveness, and the response is statistically significantly greater than the response provided by a control that does not include the dosage of the antibacterial agent. In some embodiments, the synergistic therapeutic effect of the gelseolin agent and the antibacterial agent includes enhancing the survival probability of the subject. In some embodiments, the synergistic therapeutic effect of the gelseolin agent and the antibacterial agent includes reducing microbial infections in the subject. In some embodiments, the microbial infection is a bacterial infection, optionally caused by pneumococcal species. In certain embodiments, the antibacterial agent includes a β-lactam antibiotic. In some embodiments, the antibacterial agent includes penicillin. In some embodiments, the microbial infection is caused by one type of Pseudomonas aeruginosa. In certain embodiments, the antibacterial agent is a carbapenem class antibacterial agent. In some embodiments, the antibacterial agent is meropenem. In some embodiments, the antibacterial agent includes an antifungal agent and the microbial infection includes a fungal infection. In certain embodiments, the antibacterial agent includes an antiparasitic agent and the microbial infection includes a parasitic infection. In certain embodiments, the antibacterial agent includes an antiviral agent and the microbial infection includes a viral infection.In some embodiments, the subject is a mammal, optionally a human. In some embodiments, the gelsolin agent comprises plasma gelsolin (pGSN), optionally recombinant pGSN. In some embodiments, the composition also comprises a pharmaceutically acceptable carrier. In certain embodiments, the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of the gelsolin molecule. In some embodiments, the composition also comprises a pharmaceutically acceptable carrier.

[0007] According to one aspect of the present invention, a method for increasing the therapeutic effect of an antibacterial agent against a microbial infection in a subject, the method comprising administering to the subject having a microbial infection a synergistically effective amount of each of a gelsolin agent and an antibacterial agent, wherein the administered gelsolin agent and antibacterial agent have a synergistic therapeutic effect against the microbial infection in the subject, and the synergistic therapeutic effect is greater than the therapeutic effect of the antibacterial agent administered without the gelsolin agent. In some embodiments, the antibacterial agent is administered in a clinically acceptable amount. In some embodiments, the synergistic therapeutic effect against the microbial infection is higher than the control therapeutic effect against the microbial infection, and the control therapeutic effect is the sum of the therapeutic effect of the antibacterial agent against the microbial infection and the therapeutic effect of the gelsolin agent against the microbial infection when each of the antibacterial agent and the gelsolin agent is administered without the other. In certain embodiments, the control therapeutic effect is equal to the individual therapeutic effect of the gelsolin agent. In some embodiments, the control therapeutic effect is equal to the individual therapeutic effect of the antibacterial agent administered in a clinically acceptable amount. In certain embodiments, the synergistic therapeutic effect is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the control therapeutic effect. In some embodiments, the antibacterial agent comprises an antibiotic preparation and the microbial infection comprises a bacterial infection. In some embodiments, the antibacterial agent comprises an antifungal agent and the microbial infection comprises a fungal infection. In certain embodiments, the antibacterial agent comprises an antiparasitic agent and the microbial infection comprises a parasitic infection. In some embodiments, the antibacterial agent comprises an antiviral agent and the microbial infection comprises a viral infection. In some embodiments, the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of the gelsolin molecule. In some embodiments, the gelsolin molecule is plasma gelsolin (pGSN). In certain embodiments, the gelsolin molecule is a recombinant gelsolin molecule. In some embodiments, the clinically acceptable amount of the antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent.In some embodiments, the MTD of the antimicrobial agent is the highest possible but still acceptable dosage level of the antimicrobial agent for the subject. In certain embodiments, the MTD of the antimicrobial agent is determined based at least in part on a clinically selected pre-determined toxicity limit for the antimicrobial agent. In some embodiments, the synergistic effective amount of the gelsecillin agent and the antimicrobial agent reduces the minimum effective amount (MED) of the antimicrobial agent in the subject. In some embodiments, the synergistic therapeutic effect of the administration of each of the synergistic effective amount of the antimicrobial agent and the gelsecillin agent reduces the level of microbial infection in the subject as compared to the level of control microbial infection. In some embodiments, the control infection level includes the infection level when neither the synergistic effective amount of the antimicrobial agent nor the gelsecillin agent is administered. In certain embodiments, the level of microbial infection in the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% lower than the level of control microbial infection. In some embodiments, the level of microbial infection in the subject is determined, and the means of determination includes one or more of an assay, observing the subject, assessing one or more physiological symptoms of the microbial infection in the subject, and assessing the viability of the subject. In some embodiments, the physiological symptoms include one or more of fever, malaise, and death. In certain embodiments, the physiological symptoms include lung lesions. In some embodiments, the physiological symptoms include weight loss. In some embodiments, the assay includes means for detecting the presence, absence, and / or level of characteristics of the microbial infection in a biological sample from the subject. In some embodiments, the administration of each of the synergistic effective amount of the antimicrobial agent and the gelsecillin agent increases the viability of the subject as compared to the viability of the control. In certain embodiments, the viability of the control is the viability when neither the synergistic effective amount of the antimicrobial agent nor the gelsecillin agent is administered.In some embodiments, the increase in the viability of the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the viability of the control. In certain embodiments, the administration of each of the synergistically effective amounts of the antibacterial agent and the gelsecillin agent reduces the level of lung lesions in the subject as compared to the level of lung lesions in the control. In some embodiments, the level of lung lesions in the control is the level of lung lesions in the absence of the administration of each of the synergistically effective amounts of the antibacterial agent and the gelsecillin agent. In certain embodiments, the level of lung lesions in the subject administered each of the synergistically effective amounts of the antibacterial agent and the gelsecillin agent is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% lower than the level of lung lesions in the control. In some embodiments, the subject has a Pseudomonas aeruginosa bacterial infection. In some embodiments, the antibacterial agent includes the carbapenem class and optionally includes meropenem. In certain embodiments, the bacterial infection is caused by one type of Streptococcus pneumoniae. In some embodiments, the antibacterial agent includes a β-lactam antibiotic. In some embodiments, the antibacterial agent includes penicillin. In certain embodiments, the bacterial infection is caused by one type of Pseudomonas aeruginosa. In some embodiments, the antibacterial agent is an antibacterial agent of the carbapenem class. In some embodiments, the antibacterial agent is meropenem. In certain embodiments, the bacterial infection is caused by one or more of Gram-positive bacteria, Gram-negative bacteria, Mycobacterium tuberculosis, non-tuberculous mycobacteria, spirochetes, actinomycetes, Ureaplasma species bacteria, Mycoplasma species bacteria, and Chlamydia species bacteria.In some embodiments, the means of administration of the gelsolin agent and the antibacterial agent are independently selected from oral, sublingual, buccal, intranasal, intravenous, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intra-synovial, and intraocular administration. In some embodiments, the subject is a mammal, optionally a human. In certain embodiments, the gelsolin agent is a non-therapeutic gelsolin agent. In some embodiments, the antibacterial agent is a non-therapeutic agent.

[0008] According to another aspect of the present invention, there is provided a method for synergistically treating a microbial infection in a subject, the method comprising administering to the subject having the microbial infection an effective amount of each of a gelsolin agent and an antibacterial agent, wherein the administered gelsolin agent and antibacterial agent have a synergistic therapeutic effect on the microbial infection in the subject as compared to the therapeutic effect of a control, and the antibacterial agent is administered in a clinically acceptable amount. In some embodiments, the control comprises the therapeutic effect of administering a clinically acceptable amount of the antibacterial agent administered without administering the gelsolin agent. In certain embodiments, the clinically acceptable amount of the antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent. In some embodiments, the MTD of the antibacterial agent is the highest possible but still acceptable dose level of the antibacterial agent for the subject. In some embodiments, the MTD of the antibacterial agent is determined based at least in part on a clinically selected pre-determined limit toxicity for the antibacterial agent. In some embodiments, the synergistic effective amounts of the gelsolin agent and the antibacterial agent decrease the minimum effective dose (MED) of the antibacterial agent in the subject. In certain embodiments, the MED is the lowest dose level of the antibacterial agent that provides a clinically significant response in terms of average effectiveness, and the response is statistically significantly greater than the response provided by a control that does not include the dose of the antibacterial agent. In some embodiments, the synergistic therapeutic effect is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the therapeutic effect of the control. In some embodiments, the antibacterial agent comprises an antibiotic formulation and the microbial infection comprises a bacterial infection. In certain embodiments, the antibacterial agent comprises an antifungal agent and the microbial infection comprises a fungal infection. In some embodiments, the antibacterial agent comprises an antiparasitic agent and the microbial infection comprises a parasitic infection. In some embodiments, the antibacterial agent comprises an antiviral agent and the microbial infection comprises a viral infection. In certain embodiments, the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of the gelsolin molecule. In some embodiments, the gelsolin molecule is plasma gelsolin (pGSN).In some embodiments, the gelsolin molecule is a recombinant gelsolin molecule. In certain embodiments, the synergistic therapeutic effect of the administration of each of a synergistically effective amount of an antibacterial agent and a gelsolin agent reduces the level of microbial infection in a subject as compared to the level of control microbial infection. In some embodiments, the control infection level includes the infection level when not administering each of a synergistically effective amount of an antibacterial agent and a gelsolin agent. In certain embodiments, the level of microbial infection in the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% lower than the level of control microbial infection. In some embodiments, the level of microbial infection in the subject is determined, and the means of determination includes one or more of an assay, observing the subject, evaluating one or more physiological symptoms of microbial infection in the subject, and evaluating the viability of the subject. In certain embodiments, the physiological symptoms include one or more of fever, malaise, and death. In some embodiments, the physiological symptoms include weight loss. In some embodiments, the physiological symptoms include lung lesions. In certain embodiments, the assay includes means for detecting the presence, absence, and / or level of characteristics of microbial infection in a biological sample from the subject. In some embodiments, the administration of each of a synergistically effective amount of an antibacterial agent and a gelsolin agent increases the viability of the subject as compared to the control viability. In certain embodiments, the control viability is the viability in the absence of the administration of each of a synergistically effective amount of an antibacterial agent and a gelsolin agent. In some embodiments, the increase in the viability of the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the control viability. In some embodiments, the administration of each of a synergistically effective amount of a gelsolin agent and an antibacterial agent reduces the level of lung lesions in the subject as compared to the level of control lesions.In certain embodiments, the level of the control lung lesion is the level of the lung lesion in the absence of administration of each of the synergistically effective amounts of the antibacterial agent and the gelsecillin agent. In some embodiments, the level of the lung lesion in a subject administered each of the synergistically effective amounts of the antibacterial agent and the gelsecillin agent is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% lower than the level of the control lung lesion. In some embodiments, the subject has a Pseudomonas aeruginosa bacterial infection. In certain embodiments, the antibacterial agent includes the carbapenem class and optionally meropenem. In some embodiments, the bacterial infection is caused by one type of Streptococcus pneumoniae. In certain embodiments, the antibacterial agent includes a β-lactam antibiotic. In some embodiments, the antibacterial agent includes penicillin. In some embodiments, the bacterial infection is caused by one or more of Gram-positive bacteria, Gram-negative bacteria, Mycobacterium tuberculosis, nontuberculous mycobacteria, spirochetes, actinomycetes, Ureaplasma species bacteria, Mycoplasma species bacteria, and Chlamydia species bacteria. In certain embodiments, the means of administration of the gelsecillin agent and the antibacterial agent are independently selected from oral, sublingual, buccal, intranasal, intravenous, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intra-synovial, and intraocular administration. In some embodiments, the subject is a mammal. In some embodiments, the gelsecillin agent is a non-therapeutic gelsecillin agent. In certain embodiments, the antibacterial agent is a non-therapeutic agent.

[0009] According to another aspect of the present invention, a pharmaceutical composition for use in a method of treating a subject, the composition comprising an antibacterial agent and a gelsecrin agent that synergistically increase the therapeutic effect of the antibacterial agent against a microbial infection, wherein the subject has a microbial infection, and the method comprises administering to the subject a pharmaceutical composition in an amount effective to treat the microbial infection in the subject and comprising a synergistically effective amount of each of the gelsecrin agent and the antibacterial agent, and the synergistic therapeutic effect is greater than the therapeutic effect of the antibacterial agent administered without the gelsecrin agent. In some embodiments, the gelsecrin agent and the antibacterial agent are administered to the subject separately or simultaneously. In certain embodiments, the antibacterial agent is administered in a clinically acceptable amount, and the administered gelsecrin agent and antibacterial agent synergistically enhance the therapeutic effect of administering to the subject a clinically acceptable amount of the antibacterial agent without administering the gelsecrin agent. In some embodiments, the clinically acceptable amount of the antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent in the subject. In some embodiments, the MTD of the antibacterial agent is the highest possible but still acceptable dose level of the antibacterial agent for the subject. In certain embodiments, the MTD of the antibacterial agent is determined based at least in part on a clinically selected pre-determined limit toxicity for the antibacterial agent in the subject. In some embodiments, the synergistically effective amount of the gelsecrin agent and the antibacterial agent reduces the minimum effective dose (MED) of the antibacterial agent in the subject. In some embodiments, the MED is the lowest dose level of the antibacterial agent that provides a clinically significant response in terms of average effectiveness, and the response is statistically significantly greater than the response provided by a control that does not include the dose of the antibacterial agent. In certain embodiments, the synergistic therapeutic effect of the gelsecrin agent and the antibacterial agent includes increasing the viability of the subject. In some embodiments, the synergistic therapeutic effect of the gelsecrin agent and the antibacterial agent includes reducing the microbial infection in the subject. In some embodiments, the microbial infection is a bacterial infection, optionally caused by a pneumococcal species. In certain embodiments, the antibacterial agent includes penicillin. In some embodiments, the bacterial infection is caused by one type of Pseudomonas aeruginosa. In some embodiments, the antibacterial agent is a carbapenem class antibacterial agent.In certain embodiments, the antimicrobial agent is meropenem. In some embodiments, the antimicrobial agent includes an antifungal agent and the microbial infection includes a fungal infection. In certain embodiments, the antimicrobial agent includes an antiparasitic agent and the microbial infection includes a parasitic infection. In some embodiments, the antimicrobial agent includes an antiviral agent and the microbial infection includes a viral infection. In some embodiments, the subject is a mammal. In certain embodiments, the gelsolin agent includes plasma gelsolin (pGSN), optionally recombinant pGSN. In some embodiments, the pharmaceutical composition also includes a pharmaceutically acceptable carrier. In some embodiments, the gelsolin agent includes a gelsolin molecule, a functional fragment thereof, or a functional derivative of the gelsolin molecule. In certain embodiments, the pharmaceutical composition also includes a pharmaceutically acceptable carrier.

[0010] In yet another aspect of the present invention, there is provided a method for treating viral infection in a subject, the method comprising administering to a subject having a viral infection an effective amount of a gelsolin agent, wherein the gelsolin agent is administered at least 3, 4, 5, 6, 7, 8, 9 days or more after the subject contracts the viral infection and is not administered on the day the subject contracts the viral infection, 1 day after the subject contracts the viral infection, or 2 days after the subject contracts the viral infection. In some embodiments, the effective amount of the gelsolin agent increases the therapeutic effect against viral infection in the subject as compared to the therapeutic effect of a control. In certain embodiments, the therapeutic effect of the control includes the therapeutic effect when the gelsolin agent is not administered to the subject. In certain embodiments, the antiviral agent comprises one or more of oseltamivir phosphate, zanamivir, peramivir, and baloxavir marboxil. In some embodiments, the therapeutic effect of the administered gelsolin agent is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the therapeutic effect of the control. In some embodiments, the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of the gelsolin molecule. In certain embodiments, the gelsolin molecule is plasma gelsolin (pGSN). In some embodiments, the gelsolin molecule is a recombinant gelsolin molecule. In some embodiments, the therapeutic effect of the administration of the gelsolin agent reduces the level of viral infection in the subject as compared to the level of viral infection in the control, wherein the control level of infection includes the level of infection when the gelsolin agent is not administered. In certain embodiments, the level of viral infection in the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% lower than the level of viral infection in the control.In some embodiments, the level of viral infection in a subject is determined, and the means of determination includes one or more of an assay, observing the subject, assessing one or more physiological symptoms of viral infection in the subject, and assessing the viability of the subject. In some embodiments, the physiological symptoms include one or more of fever, malaise, weight loss, and death. In some embodiments, the assay includes means for detecting the presence, absence, and / or level of characteristics of viral infection in a biological sample from the subject. In certain embodiments, administration of an effective amount of a gelseolin agent increases the viability of the subject as compared to the viability of a control. In some embodiments, the viability of the control is the viability in the absence of administration of the gelseolin agent. In certain embodiments, the increase in the viability of the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the viability of the control. In some embodiments, the means of administration of the gelseolin agent is selected from oral, sublingual, buccal, intranasal, intravenous, inhalation, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intra-synovial, and intraocular administration. In some embodiments, the subject is a mammal, optionally a human. In certain embodiments, the method also includes the step of treating the subject with an antiviral agent one or more days prior to administration of the gelseolin agent to the subject, and the antiviral agent is administered on one or more days out of the day the subject contracts a viral infection, one day after the subject contracts a viral infection, and two days after the subject contracts a viral infection. In some embodiments, a synergistically effective amount of each of the gelseolin agent and the antiviral agent is administered to the subject, and has a synergistic therapeutic effect against viral infection as compared to the therapeutic effect of the control, and the antiviral agent is administered in a clinically acceptable amount. In some embodiments, the control includes the therapeutic effect of administration of a clinically acceptable amount of an antiviral agent administered without administration of the gelseolin agent. In certain embodiments, the clinically acceptable amount of the antiviral agent is an amount less than the maximum tolerated dose (MTD) of the antiviral agent.In some embodiments, the MTD of the antiviral agent is the highest possible but still tolerable dosage level of the antiviral agent for the subject. In some embodiments, the MTD of the antiviral agent is determined based at least in part on a clinically selected pre-determined limiting toxicity for the antiviral agent. In certain embodiments, the synergistic effective amount of the gelsevirine agent and the antiviral agent reduces the minimum effective dose (MED) of the antiviral agent in the subject. In some embodiments, the MED is the lowest dosage level of the antiviral agent that provides a clinically significant response in mean effectiveness, where the response is statistically significantly greater than the response provided by a control that does not include the dosage of the antibacterial agent. In certain embodiments, the means of administration of the gelsevirine agent and the antiviral agent are independently selected from oral, sublingual, buccal, intranasal, inhalation, intravenous, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intra-synovial, and intraocular administration.

Brief Description of the Drawings

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Mode for Carrying Out the Invention

[0012] The present invention is based, in part, on the discovery that administering a gelsolin agent and an antibacterial agent to a subject having a microbial infection can result in a synergistic therapeutic effect of two agents for reducing the microbial infection. In some embodiments, the present invention, when administered to a subject having a microbial infection, acts synergistically in the subject, and these synergistic effects are greater than the therapeutic effects when either the gelsolin agent or the antibacterial agent is administered to the subject at a clinically acceptable dose and the other is not administered to the subject. The present invention includes a therapeutic composition comprising an exogenous gelsolin agent and an antibacterial agent. Certain methods of the present invention include administering a pharmaceutical composition of the present invention to a subject having a microbial infection in an amount effective to produce a synergistic therapeutic effect against the microbial infection in the subject. Some methods of the present invention include delayed dosing of a gelsolin agent to a subject having a viral infection, which enhances the treatment of the viral infection in the subject.

[0013] Synergistic therapeutic effect The methods of the present invention include producing a synergistic therapeutic effect in a subject having a microbial infection to reduce and treat the microbial infection. It was concluded that even if one or both of the gelsolin agent and the antibacterial agent do not have a statistically significant individual therapeutic effect against the microbial infection, they can be administered in combination with each other to produce a synergistic therapeutic effect against the microbial infection. Thus, in some embodiments of the present invention, microbial infections in a subject caused by a microbe resistant to one or more antibacterial agents can be effectively treated using the synergistic treatment method of the present invention due to the newly discovered synergistic therapeutic effect of administering a synergistically effective amount of a gelsolin agent and an antibacterial agent to the subject.

[0014] As used herein, the term "individual therapeutic effect" with respect to an agent such as a gelseolin agent or an antibacterial agent means the therapeutic effect of the agent when administered to a subject having a microbial infection. With respect to the methods and compositions of the present invention, the individual therapeutic effect of a gelseolin agent is the therapeutic effect on microbial infection in a subject that results from administering the gelseolin agent to the subject without administering an antibacterial agent to the subject. With respect to the methods and compositions of the present invention, the individual therapeutic effect of an antibacterial agent is the therapeutic effect on microbial infection in a subject that results from administering the antibacterial agent to the subject without administering a gelseolin agent.

[0015] As is understood in the art, a synergistic therapeutic effect is a therapeutic effect resulting from an interaction between two or more drugs, and the total therapeutic effect of the drugs is greater than the sum of the individual therapeutic effects of each drug. With respect to the method of the present invention, the total therapeutic effect of the administered gelseolin and antibacterial agent is greater than the sum of the individual therapeutic effect of the gelseolin agent and the individual therapeutic effect of the antibacterial agent. In a non-limiting example, a subject having a Streptococcus pneumoniae infection can be treated by the method of the present invention comprising the step of administering to the subject a synergistically effective amount of a plasma gelseolin (pGSN) agent and penicillin, resulting in a synergistic therapeutic effect on the infection of the subject. In this example, the therapeutic effect of administering both the pGSN agent and penicillin is greater than the sum of the individual therapeutic effect of that amount of pGSN on Streptococcus pneumoniae infection and the individual therapeutic effect of that amount of penicillin.

[0016] In some embodiments, the method of the invention comprises administering to a subject having a microbial infection a synergistically effective amount of each of a gelsolin agent and a non-therapeutic antibacterial agent. The therapeutic effect of the antibacterial agent may be enhanced by the synergistic effect of the combined administration. The term "non-therapeutic agent" as used herein refers to an antibacterial agent that does not have a statistically significant individual therapeutic effect against a microbial infection in a subject. It should be understood that the non-therapeutic agents used in connection with the methods and compositions of the invention are not antibacterial agents that are referred to in the art as "therapeutic agents" or "antibacterial therapeutic agents". For example, for a healthcare provider, an antibacterial agent that does not have a statistically significant individual therapeutic effect against a microbial infection when administered in a clinically acceptable amount would not be designated as a therapeutic agent to be administered to a subject having that microbial infection. Similarly, penicillin is recognized in the art as not having a statistically significant individual therapeutic effect against certain microbial infections, and thus penicillin would be understood and defined as a "non-therapeutic agent" with respect to those infections. In certain embodiments of the invention, the antibacterial agent is a non-therapeutic agent with respect to its individual therapeutic effect against a microbial infection in a subject. In some embodiments of the invention, the antibacterial agent is a non-therapeutic agent with respect to its individual therapeutic effect against an antibacterial agent-resistant microbial infection in a subject. A gelsolin agent lacking a statistically significant individual therapeutic effect against a microbial infection in a subject may be referred to herein as a non-therapeutic agent with respect to the microbial infection.

[0017] Individual and synergistic therapeutic effects Certain embodiments of the methods and compositions of the present invention include one or more agents that lack an individual therapeutic effect against a microbial infection in a subject. In some cases, the gelsecide agent may have an individual therapeutic effect against the microbial infection, and the antibacterial agent may not have a statistically significant individual therapeutic effect. In the case of an antibacterial agent, the lack of an individual therapeutic effect of the antibacterial agent against the microbial infection may or may not be due to antibacterial resistance of the microorganism causing the microbial infection. In connection with a microorganism or a microbial infection, the term "resistance" as used herein means a microorganism that is not killed or reduced by an antibacterial agent, respectively. In some embodiments of the present invention, the individual therapeutic effect of the antibacterial agent against the antibacterial-resistant microorganism or infection can be zero.

[0018] In certain situations, a microbial infection in a subject is due to a microorganism that is resistant to the individual therapeutic effect of an antibacterial agent. Acquired antibacterial resistance can be understood as the ability of a disease-causing microorganism to withstand exposure to an antibacterial agent that was previously an effective treatment for the disease. A microorganism that is "antibacterial-resistant" may be the cause of a microbial infection in a subject, and one or more antibacterial agents that were previously known to be therapeutically effective against the microbial infection are ineffective against the microbial infection. In a non-limiting example, a pneumococcal infection in a subject may be due to the presence in the subject of Streptococcus pneumoniae that is resistant to the therapeutic effect of one or more antibiotics.

[0019] In certain embodiments of the present invention, it will be understood that the amount of gelsolin agent administered and the amount of antimicrobial agent administered are each a clinically acceptable amount for administration to a subject. It is known that administration of an antimicrobial infection administered in a clinically acceptable amount does not reduce a particular microbial infection. For example, in some cases, the microorganism causing the microbial infection is resistant to the administered antimicrobial agent, and in other such cases, the microorganism causing the microbial infection is not sufficiently killed by administration of a clinically acceptable amount of the antimicrobial agent. In either situation, it may be possible to administer an amount of the antimicrobial agent sufficient to reduce the subject's microbial infection, but the amount required results in toxicity and / or other adverse physiological effects to the subject, so it is not a clinically acceptable amount. In contrast, the synergistic therapeutic effect of certain embodiments of the methods of the present invention enables the administration of a clinically acceptable amount of the antimicrobial agent that successfully reduces microbial infection in the subject with statistically significantly less toxicity and fewer adverse side effects in the subject.

[0020] In some embodiments of the present invention, the clinically acceptable amount of the antimicrobial agent is an amount less than the maximum tolerated dose (MTD) of the antimicrobial agent. It is understood in the art how to determine an individual's MTD in order to prevent or reduce the negative side effects of administering a drug. In some embodiments of the present invention, the MTD of the antimicrobial agent is the highest possible dose level of the antimicrobial agent for the subject that is an acceptable dose for the subject. An acceptable dose can be determined based on side effects at a given dose level, including but not limited to discomfort to the subject, physiological distress, increased risk of death of the subject, etc. In some embodiments of the present invention, the MTD of the antimicrobial agent administered to the subject is determined based at least in part on a clinically selected pre-determined limit toxicity for the antimicrobial agent. For example, the dose or amount of the antimicrobial agent effective to reduce or kill a microorganism resistant to that antimicrobial agent, when administered to a subject, may result in toxicity and / or adverse side effects that are not clinically acceptable to the subject.

[0021] The methods of the present invention are advantageous in that, due to the synergistic therapeutic effect of administering both an antibacterial agent and a gelseculin agent to a subject having a microbial infection, those methods can be used with lower doses of the antibacterial agent. In some embodiments of the methods of the present invention, the synergistically effective amounts of the gelseculin agent and the antibacterial agent reduce the minimum effective dose (MED) of the antibacterial agent in the subject. It should be understood that the amount or dose of the gelseculin agent and the amount or dose of the antibacterial agent are independently selected and are clinically acceptable amounts and doses.

[0022] In some cases, an amount of the gelseculin agent and / or an amount of the antibacterial agent has no individual therapeutic effect against the microbial infection in the subject. In some cases, an amount of the gelseculin agent and / or an amount of the antibacterial agent has an individual therapeutic effect against the microbial infection that is greater than zero. Table 1 shows the relationship between the independent therapeutic effect resulting from an amount of the gelseculin agent administered to a subject having a microbial infection, the independent therapeutic effect resulting from an amount of the antibacterial agent administered to a subject having a microbial infection, and the synergistic therapeutic effect resulting from that amount of the gelseculin agent and that amount of the antibacterial agent administered to a subject having a microbial infection. In each of the situations shown, the synergistic therapeutic effect is greater than the sum of the independent therapeutic effects of the gelseculin agent and the antibacterial agent, respectively.

[0023] [Table 1]

[0024] Therapeutic Compositions and Methods The synergistic therapeutic effect of the composition of the present invention or the treatment method of the present invention (also referred to herein as the "response" to the treatment method of the present invention) can be determined by detecting one or more physiological effects of the treatment, such as the reduction or absence of symptoms after the administration of the synergistic treatment. Additional means for monitoring and evaluating microbial infections in a subject and determining one or more of the presence, absence, level, severity, change in severity, etc. of microbial infections in a subject responsive to treatment are well known in the art, and these means may be utilized in conjunction with some embodiments of the methods described herein.

[0025] The method of the present invention comprises administering to a subject having a microbial infection a synergistic combination of a gelsecrin agent and an antibacterial agent in an amount effective to each provide a synergistic therapeutic effect for reducing the microbial infection in the subject. The gelsecrin agent and the antibacterial agent can be administered simultaneously. The gelsecrin agent and the antibacterial agent can be administered in the same formulation or in separate formulations but administered such that they are simultaneous in the subject.

[0026] The methods and compositions of the present invention can be used to treat microbial infections. As used herein, the terms "treating," "being treated," or "treatment" when used in connection with a microbial infection can refer to a prophylactic treatment that reduces the likelihood that a subject will develop a microbial infection, eliminates or ameliorates a microbial infection, prevents the microbial infection from progressing or becoming more severe, and / or refers to a treatment after a subject has developed a microbial infection to slow the progression of the microbial infection as compared to the progression of the microbial infection in the absence of the treatment method of the present invention. Gelsecrin agent Gelsolin is a highly conserved multifunctional protein that was first described in the cytosol of macrophages and later identified in many vertebrate cells (Piktel E. et al., Int J Mol Sci 2018;19:E2516; Silacci P. et al., Cell Mol Life Sci 2004;61:2614-23). The unique property of gelsolin is that its gene expresses a splice variant that encodes a distinct plasma isoform (pGSN) that is secreted into the extracellular fluid and differs from its cytoplasmic counterpart (cGSN) by an additional 25 amino acid sequence. pGSN normally circulates in mammalian blood at a concentration of 200-300 μg / ml and is one of the most abundant plasma proteins. As used herein, the term "gelsolin agent" means a composition comprising a gelsolin molecule, a functional fragment thereof, or a functional derivative of a gelsolin molecule. In some embodiments of the present invention, the gelsolin agent comprises only one or more of a gelsolin molecule, a functional fragment thereof, or a functional derivative of a gelsolin molecule. In certain specific embodiments of the present invention, the gelsolin agent may comprise one of additional additional components, which are non-limiting examples such as detectable labels, carriers, delivery agents, etc. In certain aspects of the present invention, the gelsolin molecule is plasma gelsolin (pGSN), and in certain examples, the gelsolin molecule is cytoplasmic GSN. The gelsolin molecules included in the compositions and methods of the present invention may be recombinant gelsolin molecules.

[0027] As used herein, the term "gelsolin agent" is a compound comprising an exogenous gelsolin molecule. The term "exogenous" as used herein with respect to a gelsolin molecule means the gelsolin molecule administered to a subject, even if the same gelsolin molecule, which may be referred to as an endogenous gelsolin molecule, is naturally present in the subject. The gelsolin agent included in the methods or compositions of the present invention can be a wild-type gelsolin molecule (GenBank accession number: X04412), an isoform, analog, variant, fragment, or functional derivative of a gelsolin molecule.

[0028] In some embodiments of the present invention, a "gelsolin analog" may be included, which, as used herein, refers to a compound that functions substantially similarly to either native gelsolin or a fragment thereof. A gelsolin analog includes a biologically active amino acid sequence that is substantially similar to the gelsolin sequence and may have a substituted, deleted, extended, replaced, or otherwise modified sequence with a biological activity that is substantially similar to the biological activity of gelsolin. For example, an analog of gelsolin does not have the same amino acid sequence as gelsolin but is sufficiently homologous to gelsolin to retain the biological activity of gelsolin. Biological activity can be determined, for example, by determining the properties of the gelsolin analog and / or by determining the ability of the gelsolin analog to reduce or prevent the effects of infection. Gelsolin biological activity assays are known to those of skill in the art.

[0029] Certain embodiments of the methods and compositions of the present invention include fragments of the gelsolin molecule. The term "fragment" means any portion of a gelsolin molecule that provides a segment of gelsolin that maintains at least a portion or substantially all of the level of biological activity of the "parent" gelsolin; the term means, for example, gelsolin fragments made from any source, such as naturally occurring peptide sequences, synthetic or chemically synthesized peptide sequences, and genetically engineered peptide sequences. The term "parent" as used herein with respect to a gelsolin fragment or derivative molecule means the gelsolin molecule from which the sequence of the fragment or derivative is derived.

[0030] In certain embodiments of the methods and compositions of the present invention, the gelsolin fragment is a functional fragment and retains at least a portion to all of the functions of its parent gelsolin molecule. The methods and compositions of the present invention may, in some embodiments, include "variants" of gelsolin. As used herein, a gelsolin variant can be a compound that is substantially similar in structure and biological activity to either native gelsolin or a fragment thereof. In certain aspects of the present invention, the gelsolin variant is referred to as a functional variant and retains at least a portion to all of the functions of its parent gelsolin molecule.

[0031] Gelsolin derivatives are also contemplated to be included in embodiments of the methods and compositions of the present invention. A "functional derivative" of gelsolin is a derivative that has a biological activity that is substantially similar to the biological activity of gelsolin. "Substantially similar" means an activity that can be different quantitatively but the same qualitatively. For example, a functional derivative of gelsolin can include the same amino acid backbone as gelsolin, but can also include other modifications such as post-translational modifications, for example, conjugated lipids, or covalently attached carbohydrates, depending on the need for such modifications to the performance of the therapeutic methods of the present invention. As used herein, the term also means to include chemical derivatives of gelsolin. Such derivatives may improve the solubility, absorption, biological half-life, etc. of gelsolin. The derivatives may also reduce the toxicity of gelsolin or eliminate or reduce undesirable side effects of gelsolin. Derivatives, and specifically chemical moieties that can mediate such effects, are disclosed in Remington, The Science and Practice of Pharmacy, 2012, Editor: Allen, Loyd V., Jr., 22nd Edition). Procedures for coupling such moieties to molecules such as gelsolin are well known in the art. The term "functional derivative" is intended to include "fragments", "variants", "analogs", or "chemical derivatives" of gelsolin.

[0032] Microbial infection The terms "microbe" and "microbial" are used herein to refer to microorganisms that cause diseases, which may be referred to as "microbial infections" herein. The terms microbe and microbial include, but are not limited to, microorganisms such as bacteria, fungi, viruses, and parasites, which, when present in a subject, can each cause a bacterial, fungal, viral, and parasitic infection, respectively. The term "antimicrobial agent" as used herein with respect to the treatment or reduction of an infection in a subject includes antibacterial agents, antifungal agents, antiviral agents, and antiparasitic agents that can be administered to the subject to treat a bacterial infection, a fungal infection, a viral infection, and a parasitic infection, respectively. The present invention, in some aspects, includes methods for treating an infection in a subject. In some embodiments of the present invention, the subject is known to have a microbial infection, suspected of being exposed to a microbial infection, or at risk of being exposed to or has been exposed to a microbial infection.

[0033] Characteristics of microbial infection in a subject that can be evaluated in a control subject or group include, but are not limited to, viability, death, body weight, the level of microorganisms in a biological sample from the subject, the presence of microorganisms in a biological sample from the subject, the presence, absence, and / or level of malaise, body temperature, fever, cough, pulmonary effusion, congestion, headache, chills, body aches, rash, flushing, etc. It will be understood that different microbial infections may exhibit different characteristics, and the characteristics of a microbial infection in humans may be different from those of the same microbial infection in another animal species. Characteristics present in different microbial infections as well as those present in humans and / or animals are known in the art. One of ordinary skill in the art can readily select one or more characteristics of a microbial infection for detection and evaluation in conjunction with the use of the methods and compositions of the present invention. The term "characteristic" as used herein with respect to a microbial infection may refer to the physiological symptoms of the microbial infection.

[0034] As used herein, the terms "infection" and "microbial infection" refer to disorders resulting from the invasion of a host, either superficially, locally, or systemically, by an infectious organism. Certain embodiments of the methods and compositions of the present invention can be used to treat microbial infections occurring in a subject that are caused by infectious organisms, including but not limited to microorganisms such as bacteria, viruses, parasites, fungi, and protozoa.

[0035] Microbial agent Microbial agents, which may also be referred to herein as pathogenic agents, can include bacterial agents, fungal agents, viral agents, parasitic agents, and protozoal agents. Microbial agents such as those listed below herein may, when present in a subject, cause a microbial infection in the subject.

[0036] Bacteria that may cause bacterial infection when present in a subject can include gram-negative bacteria and gram-positive bacteria. Examples of gram-positive bacteria include Pasteurella species, Staphylococcus species such as Staphylococcus aureus, Streptococcus species such as Streptococcus pyogenes of group A, the Streptococcus viridans group, Streptococcus agalactiae of group B, Streptococcus bovis, anaerobic Streptococcus species, Streptococcus pneumoniae, and Streptococcus faecalis, Bacillus species such as Bacillus anthracis, Corynebacterium species such as Corynebacterium diphtheriae, aerobic Corynebacterium species, and anaerobic Corynebacterium species, Diphtheroids species, Listeria species such as Listeria monocytogenes, Erysipelothrix species such as Erysipelothrix rhusiopathiae, Clostridium species such as Clostridium perfringens, Clostridium tetani, and Clostridium difficile.

[0037] Gram-negative bacteria include Neisseria species such as Neisseria gonorrhoeae and Neisseria meningitidis, Branhamella species such as Branhamella catarrhalis, Escherichia species such as Escherichia coli, Enterobacter species, Proteus species such as Proteus mirabilis, Pseudomonas species such as Pseudomonas aeruginosa, Pseudomonas mallei, and Pseudomonas pseudomallei, Klebsiella species such as Klebsiella pneumoniae, Salmonella species, Shigella species, Serratia species, Acinetobacter species;Species of Haemophilus, such as Haemophilus influenza and Haemophilus ducreyi, Brucella species, Yersinia species, such as Yersinia pestis and Yersinia enterocolitica, Francisella species, such as Francisella tularensis, Pasteurella species, such as Pasteurella multocida, Vibrio cholerae, Flavobacterium species, meningosepticum, Campylobacter species, such as Campylobacter jejuni, Bacteroides species, (oral cavity, pharynx) such as Bacteroides fragilis, Fusobacterium species, such as Fusobacterium nucleatum, Calymmatobacterium granulomatis, Streptobacillus species, such as Streptobacillus moniliformis, Legionella species, such as Legionella pneumophila, may be mentioned.;

[0038] Other types of bacteria include acid-fast bacteria, spirochetes, and actinomycetes. Examples of acid-fast bacteria include Mycobacterium species, such as Mycobacterium tuberculosis and Mycobacterium leprae.

[0039] Examples of spirochetes include species of Treponema, such as Treponema pallidum, Treponema pertenue, and species of Borrelia, such as Borrelia burgdorferi (Lyme disease), and Borrelia recurrentis, and species of Leptospira.

[0040] Examples of actinomycetes include species of Actinomyces, such as Actinomyces israelii, and species of Nocardia, such as Nocardia asteroids.

[0041] Viral factors that may cause viral infection when present in a subject include retroviruses, human immunodeficiency viruses including HIV-1, HDTV-III, LAVE, HTLV-III / LAV, HIV-III, and HIV-LP, cytomegalovirus (CMV), picornaviruses, polioviruses, hepatitis A virus, enteroviruses, human coxsackieviruses, rhinoviruses, echoviruses, caliciviruses, togaviruses, equine encephalitis viruses, rubella virus, flaviviruses, dengue virus, encephalitis virus, yellow fever virus, coronaviruses, rhabdoviruses, vesicular stomatitis virus, rabies virus, filoviruses, Ebola virus, paramyxoviruses, parainfluenza viruses, mumps virus, measles virus, respiratory syncytial virus (RSV), orthomyxoviruses, influenza viruses, bunyaviruses, hantaan virus, phlebovirus and nairovirus, arenaviruses, hemorrhagic fever viruses, reoviruses, orbiviruses, rotaviruses, birnaviruses, hepadnaviruses, hepatitis B virus, parvoviruses, papovaviruses, papillomaviruses, polyomaviruses, adenoviruses, herpesviruses including herpes simplex virus type 1 and herpes simplex virus type 2, varicella-zoster virus, poxviruses, smallpox virus, vaccinia virus, iridoviruses, African swine fever virus, hepatitis D virus, non-A non-B hepatitis virus, hepatitis C virus, norwalk virus, astroviruses and unclassified viruses, but are not limited thereto.

[0042] Fungal factors that may cause fungal infections when present in the subject include Cryptococcus species, such as Cryptococcus neoformans, Histoplasma species, such as Histoplasma capsulatum, Coccidioides species, such as Coccidiodes immitis, Paracoccidioides species, such as Paracoccidioides brasiliensis, Blastomyces species, such as Blastomyces dermatitidis, Chlamydia species, such as Chlamydia trachomatis, Candida species, such as Candida albicans, Sporothrix species, such as Sporothrix schenckii, Aspergillus species, and fungi causing mucormycosis, but are not limited to these.

[0043] Parasitic factors that may cause parasitic infections when present in a subject include Plasmodium species, such as Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, and Plasmodium vivax, and Toxoplasma gondii. Blood-borne parasites and / or tissue parasites include Plasmodium species, Babesia species, such as Babesia microti and Babesia divergens, Leishmania species, such as Leishmania tropica, Leishmania species, (Leishmania braziliensis, Leishmania donovani), Trypanosoma species, such as Trypanosoma gambiense, Trypanosoma rhodesiense (African sleeping sickness), and Trypanosoma cruzi (Chagas disease).

[0044] Other medically relevant microorganisms that may cause infection when present in a subject are well documented in the literature. For example, see C.G.A Thomas, Medical Microbiology, Bailliere Tindall, Great Britain, 1983, the entire contents of which are hereby incorporated by reference herein. Certain embodiments of the methods and compositions of the present invention can be used to treat infections by these and other medically related microorganisms.

[0045] Antibacterial agent Terms such as "antibacterial agent", "antibacterial drug", "antiviral agent", "antifungal agent", and "antiparasitic agent" have meanings established among those skilled in the art and are defined in standard medical texts. In summary, antibacterial agents kill bacteria or inhibit their growth or function. Antibacterial agents include antibiotics and other synthetic or natural compounds with similar functions. Antibiotics are usually low-molecular-weight molecules produced as secondary metabolites by cells, such as microorganisms. Generally, antibiotics are specific to microorganisms and interfere with the function or structure of one or more bacteria that are not present in host cells.

[0046] A major class of antibacterial agents is antibiotics. Antibiotics that are effective in killing or inhibiting a wide range of bacteria are called broad-spectrum antibiotics. Other types of antibiotics are mainly effective against classes of bacteria such as Gram-positive or Gram-negative bacteria. These types of antibiotics are called narrow-spectrum antibiotics. Other antibiotics that are effective against a single organism or disease and not against other types of bacteria are called limited-spectrum antibiotics. Antibacterial agents may be classified based on their main mechanism of action. Generally, antibacterial agents are cell wall synthesis inhibitors, cell membrane inhibitors, protein synthesis inhibitors, nucleic acid synthesis or function inhibitors, and competitive inhibitors.

[0047] Examples of antibacterial agents include, but are not limited to, aminoglycosides, β-lactam agents, cephalosporins, macrolides, penicillins, quinolones, sulfonamides, and tetracyclines. Examples of antibacterial agents include acetadapson, acetosulfone sodium, alamethicin, alexidine, amdinocillin potassium clavulanate, amdinocillin, amdinocillin pivoxyl, amiciclin, amifloxacin, amifloxacin mesylate, amikacin, amikacin sulfate, aminosalicylic acid, sodium aminosalicylic acid, amoxicillin, amphomycin, ampicillin, sodium ampicillin, apalcillin sodium, apramycin, aspartocin, astromicin sulfate, avilamycin, avoparcin, azithromycin, azlocillin, sodium azlocillin, bacampicillin hydrochloride, bacitracin, bacitracin methylene disalicylate, zinc bacitracin, bambermycin, benzoyl pascalcium, berythromycin, betamycin sulfate, biapenem, biniramycin, bifenamine hydrochloride, bispyrithione magsulfex, butikacin, butirosin sulfate, capreomycin sulfate, carbadox, carbenicillin disodium, carbenicillin indanyl sodium, carbenicillin phenyl sodium, carbenicillin potassium, carmonam sodium, cefaclor, cefadroxil, cefamandole, cefamandole nafate, cefamandole sodium, cefaparole, cefatrizine, cefazaflur sodium, cefazolin, cefazolin sodium, cefbuperazone, cefdinir, cefditoren pivoxyl, cefepime, cefepime hydrochloride, cefetecol, cefexime, cefmenoxime hydrochloride, cefmetazole, cefmetazole sodium, cefonicid monosodium, cefonicid sodium, cefoperazone sodium, ceforanide, cefotaxime, cefotaxime sodium, cefotetan, cefotetan disodium, cefotiam hydrochloride, cefoxitin, cefoxitin sodium, cefpimizole, cefpimizole sodium, cefpiramide,Cefpiramide sodium, cefpirome sulfate, cefpodoxime proxetil, cefprozil, cefroxadine, cefsulodin sodium, ceftazidime, ceftazidime sodium, cefibuten, ceftezoxime sodium, ceftriaxone sodium, cefuroxime, cefuroxime axetil, cefuroxime pivoxetil, cefuroxime sodium, cefacetrile sodium, cephalexin, cephalexin hydrochloride, cephaloglycin, cephaloridine, cephalothin sodium, cephapirin sodium, cefradine, cetocycline hydrochloride, ceftobiprole, chloramphenicol, chloramphenicol palmitate, chloramphenicol pantothenate complex, chloramphenicol sodium succinate, chlorhexidine phosphanilate, chloroxylenol, chlortetracycline bisulfate, chlortetracycline hydrochloride, cilastatin, cinoxacin, ciprofloxacin, ciprofloxacin hydrochloride, siromycin, clarithromycin, potassium clavulanate, clinafloxacin hydrochloride, clindamycin, clindamycin dextrose, clindamycin hydrochloride, clindamycin palmitate hydrochloride, clindamycin phosphate, clofazimine, cloxacillin benzathine, cloxacillin sodium, cloxyquin, colistimethate, sodium colistimethate, colistin sulfate, coumamycin, coumamycin sodium, cyclacillin, cycloserine, dalfopristin, dapsone, daptomycin, demeclocycline, demeclocycline hydrochloride, desmethacycline, denofungin, dihydrostreptomycin sulfate, dipyrithione, dirithromycin, doxycycline, doxycycline calcium, doxycycline phosphatex, doxycycline hyclate, doxycycline monohydrate, droxacin sodium, enoxacin, epicillin, epitetracycline hydrochloride, ertapenem, erythromycin, erythromycin acistrate, erythromycin estolate,Erythromycin ethylsuccinate, erythromycin gluceptate, erythromycin lactobionate, erythromycin propionate, erythromycin stearate, ethambutol hydrochloride, ethionamide, fleroxacin, floxacin, fludarabine, flumequine, fosfomycin, fosfomycin trometamol, fumoquinoline, furazolidone chloride, furazolidone tartrate, fusidic acid sodium, fusidic acid, gatifloxacin, gemifloxacin, gentamicin sulfate, gloximonam, gramicidin, haloprogin, hetacillin, hetacillin potassium, hexetidine, ibafloxacin, imipenem, isoconazole, isepamicin, isoniazid, josamycin, kanamycin sulfate, kitasamycin, levofloxacin, levoflaltadone, levopropylcillin potassium, lexicithromycin, lincomycin, lincomycin hydrochloride, linezolid, lomefloxacin, lomefloxacin hydrochloride, lomefloxacin mesylate, loracarbef, mafenide, meclocycline, meclocycline sulfosalicylic acid, megalomicin potassium phosphate, mequindox, meropenem, metacycline, metacycline hydrochloride, methenamine, methenamine hippurate, methenamine mandelate, methicillin sodium, methioprim, metronidazole hydrochloride, metronidazole phosphate, mezlocillin, mezlocillin sodium, minocycline, minocycline hydrochloride, mirincamycin hydrochloride, monensin, monensin sodium, moxifloxacin hydrochloride, nafcillin sodium, nalidixic acid sodium, nalidixic acid, natamycin, nebramycin, neomycin palmitate, neomycin sulfate, neomycin undecylenate, netilmicin sulfate, neutramycin, nifradene, nifraldazone, nifrate, nifratrone, nifurdazil, nifrimide, nifurpirinol, nifurquinazole, nifurthiazole, nitrosocycline, nitrofurantoin, nitromide, norfloxacin, novobiocin sodium, ofloxacin, ormetoprim, oxacillin sodium, oxymonam, oxymonam sodium, oxolinic acid, oxytetracyclineOxycycline calcium, Oxytetracycline hydrochloride, Pardomycin, Parachlorophenol, Paulomycin, Pefloxacin, Pefloxacin mesylate, Penamecillin, Benzathine penicillin G, Penicillin G potassium, Procaine penicillin G, Penicillin G sodium, Penicillin V, Benzathine penicillin V, Hydrabamine penicillin V, Penicillin V potassium, Pentidone sodium, Phenyl aminosalicylate, Piperacillin, Piperacillin sodium, Pirbenicillin sodium, Piridicillin sodium, Pyrlimycin hydrochloride, Pivampicillin hydrochloride, Pivampicillin pamoate, Pivampicillin probenate, Polymyxin B sulfate, Porfiromycin, Propikacin, Pyrazinamide, Zinc pyrithione, Kinderamine acetate, Quinupristin, Racephenicol, Ramoplanin, Ranimycin, Relomycin, Repromycin, Rifabutin, Rifametane, Rifamexil, Rifamide, Rifampin, Rifapentine, Rifamixin, Rolitetracycline, Rolitetracycline nitrate, Rosaramicin, Rosaramicin butyrate, Rosaramicin propionate, Rosaramicin sodium phosphate, Rosaramicin stearate, Roxithromycin, Roxarsone, Roxithromycin, Sancycline, Sanfetrinem sodium, Salmoxycillin, Sarpicillin, Scopafungin, Sisomicin, Sisomicin sulfate, Sparfloxacin, Spectinomycin hydrochloride, Spiramycin, Stallimycin hydrochloride, Steffimycin, Sterile ticarcillin disodium, Streptomycin sulfate, Streptonicodide, Sulbactam sodium, Sulfabenz, Sulfabenz amide, Sulfacetamide, Sulfacetamide sodium, Sulfacytine, Sulfadiazine,Sulfadiazine sodium, Sulfadoxine, Sulfalen, Sulfamerazine, Sulfameter, Sulfamethazine, Sulfamethizole, Sulfamethoxazole, Sulfamonomethoxine, Sulfamoxole, Sulfanilate Zinc, Sulfanitran, Sulfasalazine, Sulfisomizole, Sulfathiazole, Sulfazamet, Sulfisoxazole, Sulfisoxazole acetyl, Sulfisoxazole diolamine, Sulfomixine, Surotomycin, Sulbactam, Suncillin Sodium, Talampicillin hydrochloride, Tazobactam, Teicoplanin, Temafloxacin hydrochloride, Temocillin, Tetracycline, Tetracycline hydrochloride, Tetracycline phosphate complex, Tetroxoprim, Thiamphenicol, Tifensillin potassium, Ticarcillin cloxacillin sodium, Ticarcillin disodium, Ticarcillin monosodium, Ticaractone, Thiodonium chloride, Tobramycin, Tobramycin sulfate, Tosufloxacin, Trimethoprim, Trimethoprim sulfate, Trisulfapyrimidine, Troleandomycin, Troleandomycin sulfate, Trovafloxacin, Tyrothricin, Vancomycin, Vancomycin hydrochloride, Virginiamycin, Zorbamycin, and the like, but not limited thereto.

[0048] Antiviral agents can be isolated or synthesized from natural sources and are useful for killing viruses or inhibiting virus growth or function. Antiviral agents are compounds that prevent cell infection by viruses or virus replication within cells. There are several stages within the process of virus infection that can be blocked or inhibited by antiviral agents. These stages include virus attachment to host cells (immunoglobulins or binding peptides), virus uncoating (e.g., amantadine), synthesis or translation of viral mRNA (e.g., interferon), replication of viral RNA or viral DNA (e.g., nucleotide analogs), maturation of new viral proteins (e.g., protease inhibitors), as well as virus budding and release.

[0049] Useful antiviral agents in the present invention include, but are not limited to, immunoglobulins, amantadine, interferon, nucleotide analogs, and protease inhibitors. Specific examples of antiviral agents include acemannan; acyclovir; acyclovir sodium; adefovir; alovudine; alvircept sudotox; amantadine hydrochloride; allantoin; allyldone; atazanavir mesylate; abridin; cidofovir; cipamfylline; cytarabine hydrochloride; delavirdine mesylate; desciclovir; didanosine; disoxaril; edoxudine; envirazide; enviroxime; famciclovir; famotidine hydrochloride; fiacitabine; fialuridine; fosarilate; phosphonoformate sodium; phosphonotungstate sodium; ganciclovir; ganciclovir sodium; idoxuridine; ketoxal; lamivudine; lobucavir; memantine hydrochloride; methisazone; nevirapine; penciclovir; pyroddavir; ribavirin; rimantadine hydrochloride; saquinavir mesylate; somantadine hydrochloride; sorivudine; statolon; stubidine; tyrlolone hydrochloride; trifluridine; valacyclovir hydrochloride; vidarabine; vidarabine phosphate; vidarabine phosphate sodium; viroxime; zalcitabine; zidovudine; and zinviroxime, but are not limited thereto.

[0050] Nucleotide analogs are synthetic compounds that are similar to nucleotides but have incomplete or abnormal deoxyribose or ribose groups. Once these nucleotide analogs enter the cell, they are phosphorylated and form triphosphate forms that compete with normal nucleotides for incorporation into viral DNA or viral RNA. When these triphosphate forms of nucleotide analogs are incorporated into growing nucleic acid strands, they result in irreversible association with viral polymerases and thus cause chain termination. Examples of nucleotide analogs include, but are not limited to, acyclovir (used for the treatment of herpes simplex virus and varicella-zoster virus), ganciclovir (useful for the treatment of cytomegalovirus), idoxuridine, ribavirin (useful for the treatment of respiratory syncytial virus), didanosine, dideoxycytidine, zidovudine (azidothymidine), imiquimod, and resimiquimod.

[0051] Antifungal agents are used to treat superficial fungal infections as well as opportunistic and major systemic fungal infections. Antifungal agents are useful for the treatment and prevention of infectious fungi. Antifungal agents may be classified based on these mechanisms of action. Some antifungal agents function as cell wall inhibitors, for example, by inhibiting glucosynthase. These include, but are not limited to, basiungin / ECB. Other antifungal agents function by destabilizing membrane integrity. These include imidazoles such as clotrimazole, sertaconzole, fluconazole, itraconazole, ketoconazole, miconazole, and voriconacole, and FK463, amphotericin B, BAY38-9502, MK991, pradimicin, UK292, butenafine, and terbinafine. Other antifungal agents function by disrupting chitin (e.g., chitinase) or immunosuppression (501 cream).

[0052] Antiparasitic agents kill or inhibit parasites. Examples of antiparasitic agents useful for administration to humans, also called anthelmintics, include albendazole, amphotericin B, benznidazole, bithionol, chloroquine HCl, chloroquine phosphate, clindamycin, dehydroemetine, diethylcarbamazine acid, diloxanide furoate, eflornithine, furazolidaone, glucocorticoids, halofantrine, iodoquinol, ivermectin, mebendazole, mefloquine, meglumine antimonate, melarsoprol, metrifonate, metronidazole, niclosamide, nifurtimox, oxamniquine, paromomycin, pentamidine isethionate, piperazine, praziquantel, primaquine phosphate, proguanil, pyrantel pamoate, pyrimethamine-sulfonamide, pyrimethamine-sulfadoxine, quinacrine HCl, quinine sulfate, quinidine gluconate, spiramycin, sodium stibogluconate (sodium antimonyl gluconate), suramin, tetracycline, doxycycline, thiabendazole, tinidazole, trimethroprim-sulfamethoxazole, and trypanocide, but are not limited to these, and some of these are used alone or in combination with others.

[0053] Subject As used herein, a subject can be a vertebrate including, but not limited to, humans, mice, rats, guinea pigs, rabbits, cows, dogs, cats, horses, goats, and primates such as monkeys. In certain embodiments of the invention, the subject can be a domestic animal, a wild animal, or an agricultural animal. Thus, the present invention can be used to treat microbial infections in human and non-human subjects. For example, the methods and compositions of the present invention can be used in veterinary applications as well as in human treatment regimens. In some embodiments of the invention, the subject is a human. In some embodiments of the invention, the subject has a microbial infection and is in need of treatment.

[0054] In some embodiments, the subject already has or had a microbial infection. In some embodiments, the subject has one or more risk factors that are the cause of the infection, so the subject is at high risk of having an infection. Risk factors for microbial infection include, but are not limited to, immunosuppression, immunodeficiency, age, trauma, burns (such as thermal burns), surgery, foreign bodies, cancer, neonates, premature infants, etc. The degree of risk of acquiring a microbial infection depends on the multiplicity and severity or importance of the risk factors the subject has. Risk charts and predictive algorithms are available for assessing the risk of microbial infection in the subject based on the presence and severity of the risk factors. Other methods of assessing the risk of infection in a subject are known to those of skill in the art.

[0055] As used herein with respect to a subject having a microbial infection, the term "infected" means the day the subject was infected with a microbial infectious agent such as, but not limited to, a bacterial agent, a viral agent, a fungal agent, a parasitic agent, etc. It will be understood that the day of known or potential exposure of the subject to the microbe can be considered the zero day of infection of the subject to the microbial agent. Exposure to a microbial infection is understood to mean direct or indirect contact with an infected individual. Contact with an infected individual can be physical contact, contact with the breath, saliva, droplets, exudates, body fluids, excretions of the infected subject. In some embodiments, indirect contact can be physical contact by the subject with a substrate contaminated by an infected individual. Examples of substrates that can be contaminated by an infected individual include, but are not limited to, food, cloth, paper, metal, plastic, cardboard, fluids, air systems, etc. These and other means of exposure to a microbial infection are known in the art.

[0056] Evaluation and control Microbial infections in a subject can be detected using methods known in the art, including but not limited to, the presence of microorganisms in a biological sample obtained from the subject; the level or amount of microorganisms in a biological sample obtained from the subject; and the presence and / or level of one or more physiological symptoms of a microbial infection detected in the subject. The characteristics of a microbial infection detected in a subject can be compared to a control value for the characteristics of the microbial infection. The control value can be a predetermined value that can take various forms. The control value can be a single cut-off value, such as a median or an average value. The control value can be established based on a comparison group, such as a group of individuals with a microbial infection and a group of individuals administered a treatment for the microbial infection. Another example of a comparison group can be a group of subjects with one or more symptoms or diagnoses of a microbial infection and a group of subjects without one or more symptoms or diagnoses of a microbial infection. Of course, the predetermined value will vary depending on the particular population selected. For example, a population of individuals with a microbial infection who have been administered a gerzolin agent and not an antibacterial agent may have one or more different characteristics of the microbial infection than a population of individuals with a microbial infection who have been administered an antibacterial agent and not a gerzolin agent. Thus, the selected predetermined value can take into account the category to which the individual belongs. The appropriate category can be selected by routine experimentation by one of ordinary skill in the art.

[0057] In the method of the present invention, a control can be used to compare characteristics of different control groups, characteristics of a subject, and characteristics of a control group. Comparisons between a subject and a control, or between one control and another control, may be based on relative differences. For example, without intending to limit, the physiological symptoms of a subject treated with the synergistic treatment method of the present invention, which includes administering a gelsolin agent and an antibacterial agent to the subject, can be compared with the physiological symptoms of a control group to which the gelsolin agent has been administered and the antibacterial agent has not been administered. The comparison can be expressed in relative terms. For example, if an increase in body temperature (indicating fever) or a decrease in body temperature is characteristic of a microbial infection, the body temperature of a subject treated with the synergistic treatment method of the present invention can be compared with the body temperature level of a control. In some embodiments, a suitable control is a subject not treated with the synergistic treatment method of the present invention. The comparison between the treated subject and the control may include comparing the percentage of the temperature difference between the treated subject and the selected control. In some cases, it can be determined that the body temperature of a subject treated with the method of the present invention is lower compared to the selected control. In the comparison, the body temperature of the subject is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, or 5.9% lower compared to the body temperature level of the control, indicating.

[0058] In certain instances, it can be determined that the body temperature of a subject treated by the method of the present invention is high when compared to a selected control, where the body temperature of the subject is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, or 5.9% higher when compared to the body temperature level of the control.

[0059] In another non-limiting example, the level of microbial infection can be determined using an assay to detect the presence, absence, and / or amount of a microorganism in a biological sample obtained from a subject having the microbial infection. The results of the assay in a subject treated using the synergistic treatment method of the invention can be compared to the level of microbial infection in a control, e.g., the results of the assay on a sample obtained from a control subject not so treated. The results of the assay for assessing the level of microbial infection in a subject treated using the method of the invention can be compared to a control to determine the percentage difference between the subject level and the control level. In some embodiments, the level of microbial infection in the treated subject is less than 100% of the control infection level. In certain embodiments of the invention, the level of microbial infection in the subject to be treated is 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71%, 70%, 69%, 68%, 67%, 66%, 65%, 64%, 63%, 62%, 61%, 60%, 59%, 58%, 57%, 56%, 55%, 54%, 53%, 52%, 51%, 50%, 49%, 48%, 47%, 46%, 45%, 44%, 43%, 42%, 41%, 40%, 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1% or less of the level of microbial infection in the control.

[0060] In another non-limiting example, an increase in the level of microbial infection and / or the therapeutic effect of an antibacterial agent using the method of the present invention can be determined by comparing the viability of a control with the viability of a subject treated with the synergistic method or composition of the present invention. Non-limiting examples of the viability of a control are the viability in a subject having a microbial infection not treated by the method of the present invention. Non-limiting examples of measurable viability parameters include determination of the length of time (hours, days, weeks, etc.) that a subject continues to survive after treatment of the present invention, and determination of whether a subject dies or survives after treatment of the present invention. By comparing these and other parameters related to viability with a control, a method for evaluating and determining the therapeutic effect of the synergistic method or composition of the present invention will be understood. A non-limiting example of the viability of a control is the number of days a subject survives after treatment by the synergistic method of the present invention compared to the number of days a control survives in the absence of administration of a synergistically effective amount of an antibacterial agent and a gellosin agent. In some embodiments of the present invention, the viability of a subject treated by the synergistic method of the present invention is at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, 200%, 300%, 400%, 500% higher than the viability of the control.

[0061] In another non-limiting example, an increase in the level of microbial infection and / or the therapeutic effect of an antibacterial agent using the methods of the present invention can be determined by comparing the level of a control lung lesion with the level of a lung lesion in a subject treated with the synergistic method or composition of the present invention. Non-limiting examples of the level of a control lung lesion are the levels of lung lesions in subjects having a microbial infection not treated with the methods of the present invention. Non-limiting examples of measurable lung lesion parameters include determination of lung histopathology in a subject. In a non-limiting example, the histopathology of lung tissue (such as a sample obtained by biopsy from a subject) can be evaluated using methods known in the art, for example, the lung tissue may be observed and scored blindly by a board-certified pathologist. A scoring system can be used to compare the lung tissue of a subject to a control. In a non-limiting example, a 4-point, 4-criterion system (inflammation; infiltration; necrosis; and others, including hemorrhage) can be used to evaluate lung lesions with a maximum score of 16 points. The points for each criterion can be assigned based on no pathological findings (0), minimal (1), mild (2), moderate (3), and severe (4) pathological findings. The scoring system allows for the evaluation of lung lesions by comparing the subject tissue to the control tissue. Additional means of comparing lung lesions are known in the art and can be used in combination with the methods of the present invention. In some embodiments of the present invention, the level of a lung lesion in a subject treated with the synergistic method of the present invention is at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, 200%, 300%, 400%, 500% lower than the level of a control lung lesion.

[0062] In another non-limiting example, an increase in the level of microbial infection and / or the therapeutic effect of an antibacterial agent using the method of the present invention can be determined by comparing the level of weight loss or relative weight loss in a control with the level of weight loss or relative weight loss in a subject treated with the synergistic method or composition of the present invention. Non-limiting examples of the level of weight loss in a control are the levels of weight loss in subjects having a microbial infection not treated by the method of the present invention. Non-limiting examples of measurable weight loss and / or relative weight loss parameters include the weight of the subject before microbial infection, the weight of the subject during microbial infection before treatment by the synergistic method of the present invention, the weight of the subject after receiving the synergistic treatment method of the present invention, and the like. In a non-limiting example, the weight of a subject having a Pseudomonas aeruginosa infection can be determined before and after administration of a synergistic treatment comprising a gellin agent and a carbapenem class agent, a non-limiting example of which is meropenem. The weight of the subject can be compared with the weight of the subject before treatment, before infection, and / or the weight of another control. A decrease in weight loss in a subject after administration of the synergistic treatment of the present invention indicates a reduction in microbial infection in the subject. In some embodiments of the present invention, the level of weight loss in a subject treated by the synergistic method of the present invention is at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, 200%, 300%, 400%, 500% lower than the level of weight loss in a control.

[0063] It will be understood that the control can be a sample of a material that is tested in parallel with the experimental material, in addition to a predetermined value. Examples include samples from a control population, or control samples generated through manufacture that are tested in parallel with the experimental samples; also, the control can be a sample from a subject before, during, or after treatment according to an embodiment of the method or composition of the invention. Thus, one or more characteristics determined for a subject having an infection may later be used as a "control" value for those characteristics in that subject.

[0064] In some embodiments of the invention, the effectiveness of the synergistic method of the invention can be evaluated by comparing the synergistic treatment result in a subject treated using the method of the invention with one or both of the following: (1) the individual therapeutic effect of the gerzoline agent and (2) the individual therapeutic effect of the antibacterial agent. In certain aspects of the invention, the difference in the level of therapeutic effectiveness can be evaluated on a scale indicating an increase from the control level. In some aspects, the increase is an increase from the zero level of the control obtained in (1) or (2) to a level greater than zero resulting from treatment by the synergistic method of the invention. In some embodiments of the invention, the level of the therapeutic effect of the synergistic treatment method of the invention increases by at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, 200%, 300%, 400%, 500%, or more from the level of the therapeutic effect of the control.

[0065] Delayed dosing method Some embodiments of the present invention include a delayed dosing schedule determined to be effective in reducing viral infection in an infected subject. The therapeutic effect of the gelzoline agent is enhanced by delaying the administration of the gelzoline agent to the subject until at least 3 days or more after the day (day zero) on which the subject became infected with the virus. Some embodiments of the treatment method of the present invention include the step of administering to a subject having a viral infection an effective amount of a gelzoline agent, wherein the gelzoline agent is administered at least 3, 4, 5, 6, 7, 8, 9 days, or more after the subject having the viral infection became infected. In some embodiments, the gelzoline agent is not administered to the subject on the day (day zero) on which the subject became infected with the virus. In some embodiments, the gelzoline agent is not administered on the first day (day 1) after the subject became infected with the virus. In some embodiments, the gelzoline agent is not administered on the second day (day 2) after the subject became infected with the virus. In some embodiments of the method of the present invention, the gelzoline agent is not administered on one or more of days 0, 1, and 2 of the viral infection in the subject.

[0066] As used herein with respect to a subject having a microbial infection, the term "infected" means the day on which the subject became infected with a microbial infectious agent, e.g., but not limited to, a bacterial agent, a viral agent, a fungal agent, etc. It will be understood that the day of known or possible exposure of the subject to the microbe can be regarded as day zero of the subject's infection with the microbial agent.

[0067] Standard regimens known in the art for treating viral infections may include one or more of the following: (1) administering an antiviral agent to a subject on the day of known or potential exposure to the virus of interest; (2) administering an antiviral agent to a subject within 48 hours of known or potential exposure to the virus of interest; (3) seasonal prophylaxis with an antiviral agent by administering an antiviral agent to a subject without a specific known exposure to the virus; and (4) prophylaxis with an antiviral agent in the context of community occurrence of the virus. Exposure to a viral infection is understood to mean direct or indirect contact with an individual affected by the viral infection. Non-limiting examples of contact with an infected individual can be physical contact, i.e., contact with the breath, saliva, droplets, exudates, body fluids, excreta, etc. of the infected subject. In some embodiments, indirect contact can be physical contact by the subject with a substrate contaminated by an infected individual. Examples of substrates that can be contaminated by an individual affected by a viral infection include, but are not limited to, food, cloth, paper, metal, plastic, cardboard, fluids, air systems, etc. These and other means of exposure to a viral infection are known in the art. The methods of the present invention can be used to treat viral infections such as influenza A, B, C, and D infections. Non-limiting examples of viral infections include those caused by H1N1, H3N2, coronaviruses (e.g., 229E, NL63, OC43, HKU1, MERS-CoV, SARS-CoV, SARS-CoV-2, etc.).

[0068] A method of treating a viral infection using a timed / delayed gelsecolin agent dosing regimen may include administering a gelsecolin agent at a determined time delay following a known exposure of a subject to a viral infection, a suspected exposure to a viral infection, a potential exposure to a viral infection, and / or a risk of exposure to a viral infection. The gelsecolin agent administered may include a gelsecolin molecule, a functional fragment thereof, or a functional derivative of a gelsecolin molecule. In some embodiments, the gelsecolin molecule is plasma gelsecolin (pGSN), and in certain embodiments of the methods of the present invention, the gelsecolin molecule is a recombinant gelsecolin molecule.

[0069] In some embodiments, an effective amount of a gelsonlin agent increases the therapeutic effect against viral infection in a subject as compared to the therapeutic effect of a control, where the therapeutic effect of the control includes the therapeutic effect that occurs when the gelsonlin agent is not administered to the subject. In some embodiments, the therapeutic effect of the administered gelsonlin agent is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the therapeutic effect of the control.

[0070] In certain embodiments of the present invention, the therapeutic effect of administration of a gelsonlin agent reduces the level of viral infection in a subject as compared to the control level of viral infection, where the control infection level can be the infection level when the gelsonlin agent is not administered. In some embodiments of the present invention, the level of viral infection in the subject after administration of the gelsonlin agent in the method of the present invention is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% lower than the control level of viral infection.

[0071] One or more levels of viral infection in a subject can be determined using, for example, an assay to detect the presence, absence, and / or level of characteristics of viral infection in a biological sample obtained from the subject; observing the subject; evaluating one or more physiological symptoms of viral infection in the subject; evaluating the viability of the subject; or one or more of other means known in the art. Physiological symptoms of viral infection include, but are not limited to, one or more of fever, malaise, weight loss, and death.

[0072] Embodiments of the present invention may include administering to a subject an effective amount of a gelsevirin agent 3, 4, 5, 6, 7 days or more after or suspected exposure to a target viral infection, wherein administration of the effective amount of the gelsevirin agent increases the subject's viability compared to the viability of a control, where the control viability is the viability in the absence of administration of the gelsevirin agent. The increase in the subject's viability after administration of the gelsevirin agent using the timed dosing regimen of the present invention is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the control viability.

[0073] Delaying the time of administration of the gelsevirin agent to the subject until at least 3 days after the day (day zero) the subject became infected with the virus enhances the therapeutic effect of the gelsevirin agent and can be used in combination with administration of an antiviral agent to provide a synergistic effect between the antiviral agent and the gelsevirin agent administered to the subject. In some aspects of the present invention, the method of treating a viral infection of the present invention includes administering an antiviral agent to the subject more than 1 day prior to the delayed administration of the gelsevirin agent to the subject. In some embodiments, the antiviral agent can be administered before the subject's exposure or potential exposure to the viral infection, or on day 0, day 1, or day 2 of the subject's exposure or suspected exposure to the viral infection. It has been confirmed that depending on the effective amounts of the gelsevirin agent and the antiviral agent administered to the subject, there may be a synergistic therapeutic effect against the viral infection compared to the therapeutic effect of a control not administered in a manner that both the gelsevirin agent and the antiviral agent provide a synergistic effect. As described elsewhere herein, the antiviral agent is administered in a clinically acceptable amount, and it is understood that the control therapeutic effect can be the therapeutic effect of administration of a clinically acceptable amount of the antiviral agent administered without administration of the gelsevirin agent.

[0074] In some embodiments of the methods of the present invention, the clinically acceptable amount of the antiviral agent is an amount less than the maximum tolerated dose (MTD) of the antiviral agent. In some cases, the MTD of the antiviral agent is the highest possible, yet still acceptable, dose level of the antiviral agent for the subject. In one particular example, the MTD of the antiviral agent is determined based at least in part on a clinically selected pre-determined limit toxicity for the antiviral agent. In the methods of the present invention that include the step of administering a synergistic amount of gelsevirin agent and an antiviral agent, the synergistic effect decreases the minimum effective dose (MED) of the antiviral agent in the subject. In one particular method of the present invention, the MED is the lowest dose level of the antiviral agent that provides a clinically significant response in mean effectiveness, and the response is statistically significantly greater than the response provided by a control that does not include the dose of the antibacterial agent.

[0075] Non-limiting examples of antiviral agents that can be administered to a subject as part of an antiviral regimen are neuraminidase inhibitor antiviral drugs: oseltamivir phosphate (available as a generic version or under the trade name Tamiflu®), zanamivir (trade name Relenza®), and peramivir (trade name Rapivab®); and cap-dependent endonuclease (CEN) inhibitors: for example, baloxavir marboxyl (trade name Xofluza®).

[0076] Antiviral therapies for preventing and treating viral infections such as influenza A, B, C, and D infections are known and commonly used in the art. It is also recognized that certain viral strains can be resistant to known antiviral therapies [see, for example, Moscona, A., 2009, N Engl J Med 360;10:953-956]. Some embodiments of the methods of the present invention enhance the efficacy of antiviral agents for treating viral infections caused by viral strains that are not resistant to the antiviral agent. Certain embodiments of the methods of the present invention enhance the efficacy of antiviral agents for treating viral infections caused by viral strains that are not resistant to the antiviral agent.

[0077] In certain embodiments of the methods of the present invention, a time-limited dosage of gelsevirine regimen is administered in the absence of an antiviral agent regimen to treat viral infection. Some embodiments of the methods of the present invention treat viral infection by administering an antiviral agent regimen and a time-delayed gelsevirine regimen to a subject in need of such treatment. In some embodiments of the methods of the present invention, the administration of the antiviral agent and the delayed dosage of gelsevirine agent to a subject results in a synergistic therapeutic effect of the gelsevirine agent and the antiviral agent in the subject. The synergistic therapeutic effect of certain embodiments of the methods of the present invention can enhance the treatment of non-antiviral-resistant viral infections in a subject as compared to the control therapeutic effect. The synergistic therapeutic effect of some embodiments of the methods of the present invention can be used to enhance the treatment of antiviral-resistant viral infections in a subject as compared to the control therapeutic effect.

[0078] Preparation and Administration of Agents The methods and compositions of the present invention have important implications for the treatment of patients and also for the clinical development of new treatment methods. Here, clinical researchers are expected to use the methods of the present invention to determine the entry criteria for human subjects in clinical trials. Healthcare providers select a treatment regimen for treatment based on the net benefit expected for the subject. The net benefit is derived from the risk-to-benefit ratio.

[0079] The amount of treatment can be varied, for example, by increasing or decreasing the amount of gelsevirine agent and / or antibacterial agent administered to the subject, by changing the treatment composition administered, by changing the route of administration, by changing the timing of administration, and the like. The effective amount varies depending on factors within the knowledge and expertise of the healthcare provider, such as the particular infection or condition being treated, the age and physiological state of the subject being treated, the severity of the infection or condition, the duration of treatment, the particular route of administration, and the like. For example, the effective amount may depend on the extent to which an individual has been exposed to or affected by a microbial infection.

[0080] Effective amount As used herein in connection with the treatment methods or compositions of the present invention, the term "effective amount" is referred to as "synergistic effective amount". The method of the present invention includes the step of administering each of the gelsecrin agent and the antibacterial agent in an amount that is a synergistic effective amount of the gelsecrin agent and the antibacterial agent. When administered to a subject by the method of the present invention, the synergistic effective amount of the gelsecrin agent and the antibacterial agent results in a synergistic therapeutic effect against microbial infection in the subject and / or a reduction of microbial infection in the subject.

[0081] The effective amount is the dosage of each drug sufficient to provide a medically desirable result. Examples of drugs that can be used in certain embodiments of the compositions and methods of the present invention include, but are not limited to, gelsecrin agents and antibacterial agents. It should be understood that the drugs of the present invention are used to treat or prevent infections, i.e., they can be used prophylactically in subjects at risk of developing an infection. Thus, the effective amount is an amount that can reduce, delay, or in some cases completely prevent the onset of an infection. When a drug is used in an acute situation, it is recognized that it is used to prevent one or more medically undesirable results that normally flow from such adverse events.

[0082] The factors involved in determining the effective amount are well known to those skilled in the art and can be addressed by routine experimentation only. It is generally preferred to use the maximum dosage (alone or in combination with other therapeutic agents) of the drugs of the present invention, i.e., the highest safe dosage based on sound medical judgment. However, those skilled in the art will understand that a patient may claim a lower or acceptable dosage for medical, psychological, or virtually other reasons.

[0083] The therapeutically effective amount of the agent of the present invention is an amount effective to treat disorders such as infections. In the case of an infection, the desired response is to inhibit the progression of the infection and / or reduce the level of the infection. This may include only temporarily delaying the progression of the infection, but may also include permanently halting the progression of the infection. This can be monitored by standard diagnostic methods known to those of skill in the art. The desired response to the treatment of an infection can also be to delay the onset of the infection or to prevent the onset of the infection.

[0084] Agents and Delivery The agent used in the method of the present invention is preferably sterile and contains an effective amount of gelsecortin and an effective amount of an antibacterial agent in a weight or volume unit suitable for administration to a subject to produce the desired response. The dosage of the agent administered to the subject can be selected according to different parameters, particularly according to the mode of administration used and the condition of the subject. Other factors include the desired treatment period. In the event that the response of the subject is insufficient at the initially applied dosage, higher dosages (or different, effectively higher dosages by a more local delivery route) can be used within the tolerance of the patient. The dosage of the agent can be adjusted by an individual physician or veterinarian, particularly in the event of complications. The therapeutically effective amount usually varies from 0.01 mg / kg to about 1000 mg / kg, about 0.1 mg / kg to about 200 mg / kg, or about 0.2 mg / kg to about 20 mg / kg in once-daily or multiple-daily administrations over one or more days. The gelsecortin agent and the antibacterial agent may also be referred to herein as the agent.

[0085] Those skilled in the art are aware of various modes of administration for effectively delivering the agent of the present invention to a desired tissue, cell, or body fluid. The method and dosage administered can be adjusted by an individual physician, healthcare provider, or veterinarian, especially in the event of complications. The absolute amount administered depends on various factors including the material selected for administration, whether the administration is a single dose or multiple doses, and individual subject parameters including age, physical condition, size, weight, and the stage of the disease or condition. These factors are well known to those skilled in the art and can be addressed with only routine experimentation.

[0086] Pharmaceutically acceptable carriers include diluents, fillers, salts, buffers, stabilizers, solubilizing agents, and other materials well known in the art. Exemplary pharmaceutically acceptable carriers are described in U.S. Patent No. 5,211,657, and others are known to those skilled in the art. In certain embodiments of the present invention, such preparations may contain salts, buffers, preservatives, compatible carriers, aqueous solutions, water, etc. When used in medicine, salts may be pharmaceutically acceptable, but non-pharmaceutically acceptable salts can be conveniently used to prepare their pharmaceutically acceptable salts, which are not excluded from the scope of the present invention. Such pharmacologically and pharmaceutically acceptable salts include, but are not limited to, those prepared from hydrochloric acid, hydrobromic acid salts, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, citric acid, formic acid, malonic acid, succinic acid, etc. Also, pharmaceutically acceptable salts can be prepared as alkali metal or alkaline earth salts such as sodium, potassium, or calcium salts.

[0087] Using various modes of administration known to those skilled in the art, the pharmaceutical compositions of the present invention containing antibacterial agents and gellin agents can be effectively delivered to the subject to produce a synergistic therapeutic effect against microbial infections in the subject. Methods for administering such compositions or pharmaceutical compounds of the present invention can be topical, intravenous, oral, intracavitary, intrathecal, intrasynovial, buccal, sublingual, intranasal, transdermal, intravitreal, subcutaneous, intramuscular and intradermal administrations. In some embodiments of the present invention, the means for administering the composition of the present invention is inhalation. The present invention is not limited by the specific modes of administration disclosed herein. Standard references in the art (e.g., Remington, The Science and Practice of Pharmacy, 2012, Editor: Allen, Loyd V., Jr, 22nd Edition) provide modes of administration and formulations for delivering various pharmaceutical preparations and formulations to a pharmaceutical carrier. Other protocols useful for the administration of the therapeutic compounds of the present invention are known to those skilled in the art, and the dosage, dosing schedule, administration site, mode of administration (e.g., intra-organ), etc. are different from those presented herein. Other protocols useful for the administration of the agents of the present invention are known to those skilled in the art, and the dosage, dosing schedule, administration site, mode of administration, etc. are different from those presented herein.

[0088] For example, the administration of the agents of the present invention to non-human mammals for test purposes or for veterinary treatment purposes is carried out under substantially the same conditions as above. It will be understood by those skilled in the art that the present invention is applicable to diseases in both humans and animals. Accordingly, the present invention is intended to be used in livestock and veterinary medicine as well as in human therapy. The agent can be administered to the subject in a pharmaceutical preparation.

[0089] When administered, the pharmaceutical preparation of the present invention is applied in a pharmaceutically acceptable amount and in a pharmaceutically acceptable composition. The term "pharmaceutically acceptable" means a non-toxic substance that does not interfere with the biological activity of the active ingredient. Such preparations may usually contain salts, buffers, preservatives, compatible carriers, and optionally other therapeutic agents. When used in medicine, the salts should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts can be conveniently used to prepare their pharmaceutically acceptable salts, which are not excluded from the scope of the present invention. Such pharmacologically and pharmaceutically acceptable salts include, but are not limited to, those prepared from hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, citric acid, formic acid, malonic acid, succinic acid, etc. Also, pharmaceutically acceptable salts can be prepared as alkali metal or alkaline earth salts such as sodium, potassium or calcium salts.

[0090] The agent or composition can be combined with a pharmaceutically acceptable carrier if desired. As used herein, the term "pharmaceutically acceptable carrier" means one or more compatible solid or liquid fillers, diluents or encapsulating substances suitable for administration to humans. The term "carrier" means a natural or synthetic organic or inorganic component with which the active ingredient is combined to facilitate application. The components of the pharmaceutical composition can also be mixed with the agent of the present invention and with each other in such a way that there is no interaction that substantially impairs the desired pharmaceutical efficacy.

[0091] The pharmaceutical composition may contain a suitable buffer as described above, including acetates, phosphates, citrates, glycine, borates, carbonates, bicarbonates, hydroxides (and other bases) and pharmaceutically acceptable salts of the aforementioned compounds. The pharmaceutical composition may also optionally contain suitable preservatives: examples include benzalkonium chloride; chlorobutanol; parabens and thimerosal.

[0092] The above pharmaceutical composition may be conveniently provided in unit dosage form and may be prepared by any method well known in the pharmaceutical art. All methods include the step of bringing into association a carrier which constitutes one or more accessory ingredients with the active agent. In general, the compositions are prepared by uniformly and intimately bringing the active compound with liquid carriers, finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0093] Compositions suitable for oral administration may be presented as discrete units such as capsules, tablets, pills, troches, each containing a predetermined amount of the active compound (e.g., gelzoline). Other compositions include suspensions in aqueous liquids or non-aqueous liquids, such as syrups, elixirs, emulsions, or gels.

[0094] Pharmaceutical preparations for oral use can be obtained as solid excipients, and optionally, the resulting mixture can be comminuted and, if necessary, suitable auxiliaries can be added, and then the mixture of granules can be processed to obtain tablets or tablet cores with sugar coatings. Suitable excipients can include, in particular, fillers such as sugars including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP), etc. If desired, disintegrants such as cross-linked polyvinylpyrrolidone, agar, or alginic acid, or their salts such as sodium alginate, etc. can be added. Optionally, oral formulations can also be formulated in physiological saline or buffer solutions, i.e., in EDTA for neutralizing internal acidic conditions, or can be administered without a carrier.

[0095] Oral dosage forms of one or more of the above components are also particularly contemplated. One or more components can be chemically modified such that oral delivery of the derivative is effective. Generally, the chemical modifications contemplated are the attachment of at least one moiety to the component molecule itself, which moiety allows for (a) inhibition of proteolysis; and (b) uptake from the stomach or intestine into the bloodstream. Also desired is an increase in the overall stability of these components and an increase in the circulation time in the body. Examples of such moieties include polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone and polyproline. Abuchowski and Davis, 1981, “Soluble Polymer-Enzyme Adducts” In: Enzymes as Drugs, Hocenberg and Roberts eds., Wiley-Interscience, New York, N.Y., pp. 367-383; Newmark et al., 1982, J. Appl. Biochem. 4:185-189. Other polymers that can be used are poly-1,3-dioxolane and poly-1,3,6-tioxocane.

[0096] In the case of a medicament, the site of release can be the stomach, small intestine (duodenum, jejunum, or ileum), or large intestine. One of ordinary skill in the art has available formulations that do not dissolve in the stomach but release the substance in the duodenum or other locations in the intestine. Preferably, release avoids the deleterious effects of the gastric environment either by protection of the gellozoline agent and / or antibacterial agent or by release of the biologically active substance beyond the gastric environment such as in the intestine.

[0097] Microspheres formulated for oral administration can also be used. Such microspheres are well defined in the art. All formulations for oral administration must be in dosages suitable for such administration.

[0098] In the case of buccal administration, the composition can take the form of tablets or troches formulated by conventional methods. For administration by inhalation, the compounds for use according to the present invention can be conveniently provided in the form of an aerosol spray from a pressurized pack or nebulizer by the use of a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoromethane, carbon dioxide or other suitable gases. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. For example, gelatin capsules and cartridges for use in an inhaler or insufflator can be formulated to contain a powdered mixture of the compound and a suitable powder base such as lactose or starch.

[0099] Lung delivery of gelsolin is also contemplated herein. Gelsolin is delivered to the lungs of a mammal during inhalation and traverses the inner layer of the lung epithelium towards the bloodstream. Transnasal (or intranasal) delivery of the pharmaceutical compositions of the present invention is also contemplated. Transnasal delivery allows the pharmaceutical compositions of the present invention to pass through the bloodstream without the need to deposit the product in the lungs immediately after administering the therapeutic product to the nose. Formulations for transnasal delivery include those containing dextran or cyclodextran.

[0100] If it is desired to deliver the compound systemically, it may be formulated for parenteral administration by injection, for example, by bolus injection or continuous infusion. Injectable formulations may be presented in unit dosage form, for example, in ampoules or multi-dose containers with added preservatives. The composition can take the form of a suspension, solution or emulsion in an oily or aqueous vehicle and may contain formulating agents such as suspending, stabilizing and / or dispersing agents.

[0101] Formulations for parenteral administration include aqueous solutions of the active compound in water-soluble form. Furthermore, suspensions of the active compound can be prepared as suitable oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances such as sodium carboxymethyl cellulose, sorbitol, or dextran to increase the viscosity of the suspension. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compound and enable the preparation of highly concentrated solutions. Alternatively, the active compound may be in powder form for constitution with a suitable vehicle, for example, sterile water free from pyrogens, before use.

[0102] Agents, including but not limited to, particularly gelsevirine agents and antibacterial agents, can be provided in particles. As used herein, particles mean nanoparticles or microparticles (or in some cases larger) that can be composed of all or part of the gelsevirine or antibacterial agent described herein. The particles can contain the agent within a core surrounded by a coating including but not limited to enteric coatings. The agent can also be dispersed throughout the particle. The agent can also be adsorbed onto the particle. The particles can exhibit any order of release kinetics, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof. The particles can contain, in addition to the agent, any of the materials typically used in the pharmaceutical and medical fields, including but not limited to erosive, non-erosive, biodegradable, or non-biodegradable materials or combinations thereof. The particles can be microcapsules containing gelsevirine in solution or in a semi-solid state. The particles can be of virtually any shape.

[0103] Both non-biodegradable and biodegradable polymer materials can be used in the manufacture of particles for drug delivery. Such polymers can be natural or synthetic polymers. The polymer is selected based on the period during which release is desired. Particularly interesting bioadhesive polymers include the bioerodible hydrogels described by H.S. Sawhney, C.P. Pathak and J.A. Hubell in Macromolecules, (1993) 26:581-587, the teachings of which are incorporated herein. These include polyhyaluronic acid, casein, gelatin, gluten, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodel methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate).

[0104] The above-mentioned drugs can be included in a controlled release system. The term "controlled release" is intended to refer to any drug-containing formulation in which the method and profile of drug release from the formulation are controlled. This refers to immediate release and non-immediate release formulations, and non-immediate release formulations include, but are not limited to, sustained release and delayed release formulations. The term "sustained release" (also referred to as "extended release") is used in its conventional meaning and refers to a formulation that provides a gradual release of a drug over a long period of time and, although not necessarily, preferably results in a substantially constant blood concentration of the drug over a long period of time. The term "delayed release" is used in its conventional meaning to refer to a drug formulation in which there is a time delay between the administration of the formulation and the release of the drug therefrom. "Delayed release" may or may not be accompanied by a gradual release of the drug over a long period of time and thus may or may not be "sustained release".

[0105] The use of long-term sustained-release implants can be particularly suitable for the treatment of chronic conditions. As used herein, "long-term" release means that the implant is constructed and arranged to deliver therapeutic levels of the agent for at least 7 days, preferably 30 to 60 days. Long-term sustained-release implants are well known to those skilled in the art and include some of the above-described release systems.

[0106] The present invention also contemplates the use of kits. In some aspects of the present invention, the kit can include one or more vials of pharmaceutical preparation, a vial of pharmaceutical preparation diluent, an antibacterial agent, and a gellin agent. The vial containing the diluent for the pharmaceutical preparation is optional. The diluent vial can contain a diluent such as physiological saline for diluting a concentrated solution or lyophilized powder of the gellin agent and / or the antibacterial agent. Instructions for use can include instructions for mixing a specific amount of diluent with a specific amount of concentrated pharmaceutical preparation, thereby preparing the final formulation for injection or infusion. Instructions for use can also include instructions for treating a subject with an effective amount of the gellin agent and the antibacterial agent. The container containing this preparation can include a mark such as a conventional indicator that changes color when the preparation is autoclaved or otherwise sterilized, regardless of whether the container is a bottle, a vial with a septum, an ampule with a septum, an infusion bag, etc.

[0107] The present invention is further illustrated by the following examples, which should in no way be construed as further limitations. The entire contents of all references cited throughout this application (including literature references, issued patents, published patent applications, and co-pending patent applications) are hereby expressly incorporated herein by reference.

[0108] The following examples are provided to illustrate specific examples of the practice of the present invention and are not intended to limit the scope of the present invention. As will be apparent to those skilled in the art, the present invention will find uses in various compositions and methods.

Examples

[0109] Example 1 Antibiotic-resistant pneumococcal pneumonia can occur and may be a problem. Studies have been conducted to evaluate a novel treatment strategy for combating infection, including means to enhance innate immunity. Experiments were performed to determine the effect of pGSN administration on macrophages and host survival. Methods: Bacterial species and cultures S. pneumoniae serotype 3 (catalog number 6303, American Type Culture Collection, Rockville, MD) was cultured overnight on 5% sheep blood-enriched agar Petri dishes (catalog number 90001-282, VWR, West Chester, PA), prepared, and quantified as previously reported (Yang Z. et al., Am J Physiol Lung Cell Mol Physiol 2015;309:L11-6). In vitro and in vivo procedures (1) In vitro study An in vitro study was performed to determine the survival rate of bacteria by adding 125 - 250 μg / ml of pGSN to the bacterial culture. (2) In vivo study B16 mice were challenged with 10 5 pneumococci by i.n. instillation and 10 mg of pGSN was administered s.c. 2 hours before and 8 hours and 20 hours after infection. In some studies, pGSN was administered as an aerosol 15 or 30 minutes before infection. The aerosol was generated using a 5 mg / ml solution as described by Hamada, K. et al., J. Immunology. 2003;170(4):1683-9.

[0110] Results / Discussion The results of in vitro studies showed that pGSN improved macrophage uptake (Figure 1A) and killing of internalized pneumococci (Figure 1B) when present at 125 - 250 μg / ml, which is similar to normal plasma levels. In vivo, pGSN (10 mg s.c. 2 hours before and 8 and 20 hours after infection, and 10 5 improved bacterial clearance in Bl6 mice challenged with pneumococci (fewer viable bacteria at 24 hours) (Figure 1C); similar results were seen when pGSN was administered as an aerosol for 15 or 30 minutes before infection; the aerosol was generated using a 5 mg / ml solution as described in Hamada, K., et al., J. Immunology. 2003;170(4):1683 - 9 (Figure 1D). Systemic pGSN (s.c.) improved survival in primary (Figure 1E, using 3×10 5 CFU inoculation) or secondary pneumococcal pneumonia following influenza (Figure 1F, using 500 CFU inoculation 7 days after mild influenza infection with PR8) even without antibiotic treatment. *=p <.05 vs control, n = 6 - 12 per group. Serotype 3 Strep. Pneumoniae [ATCC #6303] was used in all experiments.

[0111] Macrophage NOS3 is an important mechanism of host defense against pneumonia in mice and also functions in human macrophages (Yang, Z., et al., Elife. 2014;3. Epub 2014 / 10 / 16. Doi 10.7554 / elife.03711). The results showed that this pathway functions as an important mechanism of the pGSN effect on macrophages because pGSN was unable to improve the bactericidal response in NOS3 - deficient macrophages (Figure 2A) and NOS3 - deficient mice (Figure 2B).

[0112] Additional studies were performed using Escherichia coli and Francisella tularensis (see Yang, Z. et al., American Journal of Physiology Lung Cellular and Molecular Physiology. 2015;309(1):L11-6).

[0113] Example 2 A study was performed to evaluate the effect of pGSN treatment on an antibiotic susceptibility and antibiotic resistance mouse model of pneumococcal pneumonia. Methods: Bacterial species and cultures S. pneumoniae serotypes 3 and 14 (catalog numbers 6303 and 700677, respectively) were obtained from the American Type Culture Collection (Rockville, MD). Bacteria of serotype 3 were cultured overnight on 5% sheep blood supplemented agar Petri dishes (catalog number 90001-282, VWR, West Chester, PA), prepared, and quantified as previously reported (Yang Z. et al., Am J Physiol Lung Cell Mol Physiol 2015;309:L11-6). Serotype 14 required a more detailed protocol to achieve consistent results, so the growth protocol reported by Restrepo AV et al., BMC Microbiol 2005;5:34 was followed, which uses two consecutive growths in liquid broth culture before centrifugation and adjustment of the bacterial concentration by OD600 for in vivo administration.

[0114] Mouse model of pneumococcal pneumonia Normal 6 - to 8 - week - old male CD1 mice were obtained from Charles River Laboratories (Wilmington, MA). As previously reported, primary pneumococcal pneumonia was induced (Yang Z. et al., Am J Physiol Lung Cell Mol Physiol 2015;309:L11 - 6). For antibiotic - susceptible pneumonia, intranasal instillation of 1.5 - 2×10 6 colony - forming units (CFU) of Streptococcus pneumoniae type 3 was performed in mice under anesthesia with ketamine (72 mg / kg ip) plus xylazine (9.6 mg / kg ip). Streptococcus pneumoniae type 14, which is resistant to penicillin (minimum inhibitory concentration (MIC) = 8 μg / ml) and other antibiotics (Jabes D. et al., J Infect Dis 1989;159:16 - 25), was used to model antibiotic - resistant pneumonia. For this pathogen, a highly lethal inoculum of approximately 300×10 6 colony - forming units (CFU) used for intranasal instillation under anesthesia as described above was identified. In most experiments, 10 mice per group were used in the vehicle, penicillin (PEN), pGSN, or PEN + pGSN groups.

[0115] Treatment and Outcome Recombinant human pGSN (rhu-pGSN) was synthesized in Escherichia coli and purified by Fujifilm Diosynth (Billingham, UK). rhu-pGSN was administered to mice by intraperitoneal injection at a dose in the range of 5 - 10 mg as detailed in the results. In some experiments, penicillin (procaine G injection suspension, NDC 57319-485-05, Phoenix Pharmaceuticals) was administered intramuscularly by i.m. injection at 0.1 - 2 mg. Mice were monitored for 10 days, and composite indices (i.e., rounded back appearance, ruffled fur, or partially closed eyes each scored 1 point; penile protrusion or spread hindquarters scored 1.5 points; lethargy scored 2 points; maximum score was 8; the assessment was performed without blinding the treatment groups) in accordance with the guidelines of Burkholder T. et al., Curr Protoc Mouse Biol 2012;2:145-65 were used to measure survival rate, change in body weight, and overall morbidity. For animals that did not survive, the body weight and morbidity score on the last day of survival were carried forward. To evaluate lung inflammation by quantifying neutrophil influx, one cohort of animals received lung lavage 48 hours after sacrifice from infection as described above (Yang Z. et al., Am J Physiol Lung Cell Mol Physiol 2015;309:L11-6, and Yang Z. et al., Elife 2014;3). After centrifugation, the resuspended lavage fluid samples were counted using a hemocytometer, and the difference in cell numbers was counted in cytocentrifuge preparations stained with Wright-Giemsa.

[0116] Statistical Analysis Data were analyzed using Prism (GraphPad Software) or SAS (SAS Institute) software. Differences in Kaplan-Meier survival curves were analyzed using the log-rank test with Sidak adjustment for multiple comparisons. For other measurements, differences between groups were investigated by ANOVA.

[0117] Results The delayed treatment with rhu-pGSN was tested in the same mouse model previously used to demonstrate the improvement in survival with pretreatment (Yang Z. et al., Am J Physiol Lung Cell Mol Physiol 2015;309:L11-6). As shown in Figure 3A, pGSN treatment given only on days 2 and 3 after serotype 3 pneumococcal infection significantly improved survival from a lethal inoculum compared to vehicle control, even in the absence of antibiotic treatment. In contrast to subsequent experiments using serotype 14, 100% survival of antibiotic-treated mice confirmed that serotype 3 is highly sensitive to penicillin (Figure 3B).

[0118] To determine whether these findings extend to antibiotic-resistant pneumonia, a similar model was developed using highly virulent serotype 14 pneumococci. Treatment was initiated 24 hours after infection and continued daily for 9 days. Mice treated with diluent vehicle alone experienced high mortality (Figure 4A). Penicillin treatment alone was ineffective (Figure 4A), consistent with the reported high level of in vitro resistance of this strain (Jabes D. et al., J Infect Dis 1989;159:16-25).

[0119] During the 24 hours prior to treatment, all mice experienced the same degree of deterioration as evidenced by equivalent weight loss and morbidity scores. Neutrophil influx at 48 hours after infection was decreased in animals treated with a single dose of pGSN regardless of the presence or absence of penicillin (total lavage neutrophils × 10E4 for vehicle, PEN, pGSN, and PEN + pGSN groups respectively: 186 ± 54, 153 ± 74, 111 ± 16, 104 ± 20; p <.03, n = 5-6 / group). Treatment with rhu-pGSN alone resulted in a significant improvement in overall survival, recovery from weight loss, and improvement in morbidity scores (Figures 4A-C).

[0120] In vitro, penicillin treatment alone or in combination with pGSN did not affect bacterial growth (increase in bacterial CFU, vehicle, PEN (16 μg / ml) or PEN + pGSN (250 μg / ml) for 1 hour (h) of culture: 88000, 105000, 88000, respectively, average of 2 repeats).

[0121] In vivo, treatment with the combination of penicillin and pGSN resulted in a higher survival rate than treatment with pGSN alone (Figure 4A), but this was not statistically different when adjusted for multiple comparisons (p = 0.47, see Figure 5). The results of all survival experiments are shown in Figure 5, which shows that for each of the 9 experiments, the survival rate was highest in the pGSN + PEN group, followed by pGSN alone compared to either PEN or vehicle alone (the combination of pGSN + PEN is significantly better than pGSN alone). The table in Figure 5 provides the results from 9 experiments in which 4 treatments of delayed administration were tested. Data from the final 4 experiments, which used essentially the same treatments and are representative of the overall results obtained in all 9 studies, are shown in Figures 4A - C. Figure 5 provides details of all 9 experiments, including pilot and rangefinder tests. Column H shows the changes in the bacterial growth method for the excellent growth results obtained using the double - growth method in BHI broth of penicillin - resistant pneumococci (Restrepo AV et al., BMC Microbiol 2005;5:34). The difference in survival rates was statistically significant as determined by the analysis of all 9 pooled studies using log - rank analysis with Sidak correction for multiple comparisons. The details of the results of the statistical analysis of the final 4 experiments (numbers 6 - 9) are summarized in Figures 4A - C.

[0122] Discussion The study was designed and configured to mimic the clinical situation shown by the subjects after the infection became apparent. Thus, the experiment was conducted using a clinically relevant scenario of delaying administration until the mice became visibly ill, rather than using pretreatment or co-treatment used in previous studies (Yang Z. et al., Am J Physiol Lung Cell Mol Physiol 2015;309:L11-6). Using this design, the potential of pGSN to improve treatment outcome was evaluated. The key finding was that survival was improved by the delay in pGSN treatment when used alone without antibiotics or in combination with the next best antibiotic against which the bacterial species was highly resistant. The decrease in bronchoalveolar neutrophil numbers observed in infected pGSN-treated animals may reflect accelerated bacterial clearance by pGSN-stimulated resident macrophages, the anti-inflammatory activity of pGSN, or both. For serotype 14, the ability to study longer delays before treatment was limited in the pilot study because there were relatively numerous deaths by day 2 or 3 without treatment. Studies are conducted to investigate other antibiotic-resistant bacteria in other model systems. Previous findings that pGSN enhances the bactericidal function of macrophages against other bacteria (e.g., E. coli, F. tularensis LVS [Yang Z. et al., Am J Physiol Lung Cell Mol Physiol 2015;309:L11-6.]) are promising in this regard but direct testing is needed.

[0123] The overall data suggest a synergistic interaction between pGSN and penicillin treatment that was ineffective by itself. However, this conclusion depends on not only the final test performed but also the analysis of all pooled distance measurements. Analyzing only the final four repeated studies (Figures 4A - C), the comparison is in the same direction but statistical significance is not achieved. Enhancement of the penicillin effect on in vitro bacterial growth by the accompanying rhu - pGSN was not observed. While not intending to be bound by a particular theory, these data suggest that antibacterial defense enhanced by pGSN may be more effective against bacteria slightly disrupted (but not killed) by penicillin. This mechanism may merit future attention, especially if similar results are observed in other infections caused by resistant bacteria. In summary, rhu - pGSN can improve the outcome in a highly lethal pneumococcal pneumonia model when given after a clinically appropriate delay, even in the setting of antibacterial resistance. These findings support further evaluation of pGSN as adjunctive therapy for severe antibiotic - resistant infections.

[0124] Example 3 A study was conducted to evaluate the effect of rhu - pGSN treatment on meropenem in a highly lethal multidrug - resistant Pseudomonas aeruginosa pneumonia in a neutropenic mouse model. Methods Generation of rhu - pGSN Recombinant human plasma gelsolin (rhu - pGSN) was produced in Escherichia coli and then lyophilized for reconstitution. A vehicle control containing formulation components was used for comparison mice. Bacterial species and growth conditions Pseudomonas aeruginosa UNC-D is sputum isolated from a patient with cystic fibrosis [Lawrenz MB et al., Pathog. Dis. 73 (2015)]. The bacteria were cultured on trypticase soy agar (TSA) plates and in Lennox broth with shaking as a broth culture at 37 °C. The minimum inhibitory concentrations of the UNC-D strain are ceftazidime [32 μg / ml], meropenem [8 μg / ml], imipenem [16 μg / ml], tobramycin [32 μg / ml], piperacillin [16 μg / ml], aztreonam [4 μg / ml], colistin [1 μg / ml], and fosfomycin [256 μg / mL]. The bacteria were cultured overnight in Lennox broth, washed with 1× PBS, and then diluted to the final concentration based on the OD 600 base estimated value and the final delivery volume of 50 μl for preparation for animal challenge studies. Bacterial inoculation was confirmed by serial dilution on TSA plates and colony counting.

[0125] Animal respiratory infection model The BALB / c infection model of the Pseudomonas aeruginosa UNC-D strain [Lawrenz MB et al., (2015) Pathog. Dis. 73 (5): ftv025] was specifically designed to test adjuvant therapies that may improve the efficacy of meropenem monotherapy, which failed against the multidrug-resistant (MDR) Pseudomonas aeruginosa UNC-D strain that is resistant to several clinically important antibiotics including meropenem. Previous experience has demonstrated that this model is most beneficial when investigating new compounds using meropenem doses that provide approximately 50% mortality with meropenem treatment alone [Lawrenz MB et al., (2015) Pathog. Dis. 73 (5): ftv025]. Mice were housed and treated according to the standard animal experimental guidelines of the University of Louisville. Briefly, female BALB / c mice were neutropenized using cyclophosphamide injections (150 mg / kg) on days -5 and -3 before infection, and usually, the neutrophil count decreased by approximately 90%. Approximately 105.5 The UNC-D of CFU was directly instilled into the lungs by intratracheal injection via a cannula. Meropenem (Hospira; Lake Forest, IL) was administered by subcutaneous injection starting 3 hours after infection and every 8 hours for 5 days.

[0126] To determine whether rhu-pGSN adjuvant therapy improves the efficacy of meropenem, 12 mg / day of rhu-pGSN was administered by intraperitoneal injection of 0.3 ml at -24, -3, 3, 27, 51, 75, 99, and 123 hours after infection. The morbidity of the mice was monitored every 8 hours for 7 days after infection, which also included the temperature measured via a transponder (BioMedic Data Systems; Seaford, DE) implanted subcutaneously before the start of the study. Moribund mice were humanely euthanized and scored as succumbing to infection at the next time point. As described previously, tissue samples were taken for bacterial counts and pathology [Lawrenz MB et al., Pathog. Dis. 73 (2015)]. Mice that survived until 7 days were scored as having withstood the infection and euthanized; the tissues were processed similarly. The histopathology of the lungs was scored blindly by a board-certified veterinary pathologist. A 4-point, 4-criterion system (inflammation; infiltration; necrosis; and others, including hemorrhage) was used to evaluate the lung lesions, with a maximum score of 16 points. The points for each criterion were assigned based on no pathological findings (0), minimal (1), mild (2), moderate (3), and severe (4) pathological findings.

[0127] Statistical Analysis Overall, three equivalent experiments were independently performed using this model. Titration experiments were performed using a new batch of meropenem to determine the effective dose (ED) of each lot of the antibiotic before the formal experiment 50This was done when estimating. The overall survival period and the survival period with minimal lung injury (post hoc defined as histopathological score ≤2) were tabulated for experimental conditions in which the entire experiment and the control group with meropenem alone protected ≤50% of the mice. 95% confidence intervals and p-values for the difference in the proportion of surviving mice between treatment groups with and without rhu-pGSN were calculated via normal approximation of the binomial distribution. For individual experimental conditions in which the mortality rate in the control meropenem group was approximately 50% or more, survival curves were analyzed by the log-rank test, temperature data were analyzed by two-way ANOVA, and bacterial load and pathological scores were analyzed by one-way ANOVA using Tukey's post hoc test for multiplicity adjustment. The predefined primary endpoint was the survival rate 7 days after the infection challenge. During the analysis of these data, a "survival plus" endpoint for investigating the survival rate in healthy lungs (histopathological score ≤2) was used as a clinically significant extension of the favorable outcome. Bacterial load and temperature response were not included in this two-pronged composite because they were not direct measures of clinical improvement.

[0128] Results rhu-pGSN improved the survival rate of mice infected with Pseudomonas aeruginosa To determine whether rhu-pGSN could improve the efficacy of meropenem against lung infection, female BALB / c mice were neutropenized with cyclophosphamide (n = 8), infected with MDR Pseudomonas aeruginosa, and treated with various doses of meropenem until meropenem therapy began to fail in this model (i.e., the ED of meropenem 50(Approaching) the dosage was determined. Mice were treated with the selected dosage of meropenem with or without rhu-pGSN for 5 days after infection and monitored for the onset of fatal disease for 7 days after infection (Figure 6). In both Experiments 1 and 2, treatment with 1250 mg / kg / day of meropenem resulted in a survival rate of ≤50%, indicating the failure of meropenem treatment, which enabled confirmation of whether adjuvant therapy with rhu-pGSN could improve efficacy. Focusing on the animals that received this dosage, the addition of rhu-pGSN numerically increased the number of animals that survived until the end of each study (Figures 7A - B). Combining the two consecutive studies, 31% of the mice that received meropenem alone survived for 7 days, compared with a survival rate of 75% when meropenem was administered to mice with rhu-pGSN (Δ(95% confidence interval) = 44% (13, 75); p = 0.0238; Figure 7C). In a third experiment using a different lot of meropenem that showed higher-than-expected meropenem efficacy (75% survival rate in the meropenem-only group), no difference in survival rate was observed between the treatment groups (Figure 6).

[0129] To confirm whether the increased survival rate by rhu-pGSN therapy is associated with a decrease in the bacterial load in the lungs, the colony count was determined from the lungs of mice administered 1250 mg / kg / day at the time of euthanasia (Figures 8A - C). A general trend was observed suggesting that rhu-pGSN improved the control of the bacterial load in the lungs of infected mice compared with meropenem alone, but a statistically significant difference in the number of bacteria was observed only in the second study (p = 0.0273).

[0130] The overall survival periods of all dosing groups in the three combined experiments were 35 / 64 (55%) and 46 / 64 (72%), respectively, in mice treated with meropenem with or without rhu-pGSN [Δ (95% confidence interval) = 17% (1, 34)]. Treatment with adjunctive rhu-pGSN enhanced the efficacy of meropenem against pulmonary infection by Pseudomonas aeruginosa, but inhibition of bacterial growth in the lungs may only partially explain the observed effects. Interestingly, in both studies, spread from the lungs to the spleen was controlled by meropenem alone, but it was observed that pGSN enabled colonization of the spleen in some animals. This observation, which was not significant in either study alone, was demonstrated to significantly increase the number of spleens in pGSN-treated mice when the data were combined. Together with the improved survival rate, these observations were consistent with rhu-pGSN exerting an opsonizing effect to enhance spleen uptake.

[0131] rhu-pGSN limits acute lung injury Since there was no clear relationship between the reduction of lung bacterial load and the increased survival rate of mice receiving rhu-pGSN, the likelihood that rhu-pGSN protection is mediated by alternative or additional mechanisms increased. Since pGSN modulates inflammation, the question of whether lung injury was reduced by rhu-pGSN adjuvant therapy was investigated in animals infected with Pseudomonas aeruginosa receiving 1250 mg / kg / day. Representative sections of lung tissue taken from the animals were blindly scored for pathology by a board-certified veterinary pathologist. Adding rhu-pGSN to meropenem reduced host lung injury (Figs. 9A - B; p = 0.0035 and p = 0.1514, respectively). Combining the data from these two independent studies, the mean pathology score of mice receiving meropenem alone was 6.86, while the mean pathology score of mice receiving both meropenem and rhu-pGSN was 2.53 (Fig. 9C; p = 0.0049).

[0132] Based on these observations that rhu-pGSN provided protection from lung injury, the analysis was extended to include mice receiving meropenem doses above and below 1250 mg / kg / day. The overall survival of mice receiving different doses of meropenem in three separate experiments is shown in Figure 6. Animals that survived infection for 7 days were grouped as showing either nearly normal lung histology (pathology score ≤2) or signs of lung lesions (pathology score >2). Using this criterion retrospectively, the overall survival with mild lung injury was seen in 26 / 64 (41%) mice receiving meropenem alone and 38 / 64 (59%) mice receiving meropenem plus rhu-pGSN [Δ (95% confidence interval) = 19% (2, 36)] (Figure 10). To exclude the noise generated by very effective and ineffective meropenem doses, arbitrarily but clinically reasonable exclusion limits of ≥75% and ≤25% were imposed on the control survival rate. At this intermediate point of responsiveness to meropenem alone, another exploratory post hoc analysis showed favorable outcomes (survival with nearly normal lungs) in 12 / 32 (37.5%) cases with meropenem alone and 27 / 32 (84.4%) cases with the combination of meropenem and rhu-pGSN. [Δ = 47% (26, 68)].

[0133] Using the surviving mice as the denominator, nearly normal lung histopathology was seen in 26 / 35 (74.3%) and 38 / 46 (82.6%) for monotherapy with meropenem versus combination therapy with meropenem and rhu-pGSN, respectively. Combining these data indicates that the addition of rhu-pGSN may reduce lung injury caused by Pseudomonas aeruginosa infection treated with antibacterial agents alone.

[0134] Plasma gelsolin accelerates the resolution of the host's systemic response As part of monitoring the progression of the disease, the body temperature of the host was tracked during the course of infection. In this model, all mice tended to show a gradual decrease in body temperature within the first 24 hours after infection. In the case of mice that received effective treatment, the temperature eventually returned to normal, but the temperature of mice that received less effective treatment continued to decline [Lawrenz MB et al., (2015) Pathog. Dis. 73(5):ftv025]. By observing the time course of temperature normalization, the difference in recovery rates between different treatments could be evaluated. In these experiments, the dosing regimen approaching the target ED of meropenem alone was focused on, and the question of whether pGSN promoted the recovery of thermoregulation in mice resistant to infection was investigated. In two studies that achieved a survival advantage, mice typically had a body temperature decrease of approximately -12.22°C (10°F) within the first 24 hours after infection (Figures 11A - D). Mice treated with meropenem alone, which were expected to survive until day 7, began to recover their body temperature to 35°C (95°F) within 3 - 5 days after infection. In contrast, the recovery of the host's body temperature was much more rapid in mice treated with rhu - pGSN and meropenem, and the body temperature of the survivors had recovered to 35°C (95°F) by day 2. Therefore, in addition to improving survival rate and lung lesions, the adjunctive rhu - pGSN accelerated the host's overall recovery as measured by the temperature curve. In a third experiment where no effect of rhu - pGSN on survival was seen, no difference in the temperature course was observed between treatment groups. 50 Focused on the dosing regimen approaching the target ED of meropenem alone, the question of whether pGSN promoted the recovery of thermoregulation in mice resistant to infection was investigated. In two studies that achieved a survival advantage, mice typically had a body temperature decrease of approximately -12.22°C (10°F) within the first 24 hours after infection (Figures 11A - D). Mice treated with meropenem alone, which were expected to survive until day 7, began to recover their body temperature to 35°C (95°F) within 3 - 5 days after infection. In contrast, the recovery of the host's body temperature was much more rapid in mice treated with rhu - pGSN and meropenem, and the body temperature of the survivors had recovered to 35°C (95°F) by day 2. Therefore, in addition to improving survival rate and lung lesions, the adjunctive rhu - pGSN accelerated the host's overall recovery as measured by the temperature curve. In a third experiment where no effect of rhu - pGSN on survival was seen, no difference in the temperature course was observed between treatment groups.

[0135] Discussion Adding rhu-pGSN to meropenem in an established mouse model of severe multidrug-resistant Pseudomonas aeruginosa pneumonia improved survival. Normalization of body temperature in surviving mice generally occurred more rapidly with rhu-pGSN adjunctive therapy than with meropenem alone. Lungs from rhu-pGSN recipients generally had fewer viable bacteria. Furthermore, rhu-pGSN reduced the extent of acute lung injury in surviving animals, which may represent a clinically important advance in the treatment of severe bacterial pneumonia. Collectively, these findings suggest that the survival benefit conferred by adding rhu-pGSN to meropenem treatment is likely due in large part to the rhu-pGSN-mediated reduction in bacterial burden and severity of lung injury during the infectious process.

[0136] The first line of host defense against infection is accompanied by an intense inflammatory response. However, excessive local and systemic inflammation can be harmful to vital organs proximal and distal to the primary site of infection. As acute injury resolves, pGSN promotes resolution of the inflammatory process and limits resulting damage.

[0137] The putative effect of adding rhu-pGSN treatment to meropenem was examined in a highly lethal multidrug-resistant Pseudomonas aeruginosa pneumonia in a neutropenic mouse model. All mice died within approximately 24 hours of infection without immediate antimicrobial therapy. Treatment with rhu-pGSN alone extended the mean survival time by approximately 12 hours. Titration experiments were performed with each batch of antibiotic to determine the dose of meropenem that produced a mortality of ≧50%. Nevertheless, the outcome was not always predictable, and in some trials the mortality of the meropenem control was ≦25% or ≧75%. Under such extreme conditions, the putative effect of adjunctive rhu-pGSN on the outcome may be masked because the mice are either too sick or not sick enough. Nevertheless, rhu-pGSN given with meropenem was more effective than meropenem alone under most conditions.

[0138] These pre - clinical data further strengthen the growing evidence that rhu - pGSN, as an adjunct to standard treatment modalities, may be effective in increasing survival while limiting lung injury. Even at supra - physiological levels throughout the dosing interval, no severe adverse events or drug - related adverse events were observed in rhu - pGSN recipients treated continuously for 3 days.

[0139] Using an established model of murine Gram - negative pneumonia, there was considerable variability in the number of bacterial colonies from bronchoalveolar lavage and the histopathological lung injury score at euthanasia within and between experiments, but these were higher in mice receiving meropenem alone compared to mice treated with meropenem and rhu - pGSN. Most notably in situations where meropenem alone was relatively ineffective, adding rhu - pGSN to meropenem reduced both mortality and parenchymal injury.

[0140] Example 4 Method Mouse model of influenza Normal 6 - to 8 - week - old male CD1 mice were obtained from Charles River Laboratories (Wilmington, MA). Due to budget and time constraints, only male mice were used. All mice arrived and were co - housed 1 week prior to the start of the experiment. In each trial, individual batches of mice were used. The mouse - adapted strain of H1N1 influenza virus, A / Puerto Rico / 8 / 1934 (PR8), quantified as plaque - forming units (PFU), was procured from ViraSource (Durham, NC). Mice were anesthetized by intraperitoneal injection of 72 mg / kg ketamine plus 9.6 mg / kg xylazine. Next, the mice received an intranasal inoculation of 25 μl of a PBS suspension containing the virus (ranging from 400 to 1000 PFU depending on the trial) or vehicle alone. All infections were performed at approximately the same time (around 10:00 am). In the first titration, 400 PFU was identified as the dose that produced approximately 60% mortality in vehicle - treated mice, and this dose was used in most of the trials (see Figure 12). In most trials, at least 10 mice per group were used for the vehicle and pGSN - treated groups; details of the influenza dose, total number of mice, and their body weights are provided in the underlying data tables [Kobzik L: "Expanded Tables 1 & 2". Harvard Dataverse, V1 2019. www.doi.org / 10.7910 / DVN / 53GJY1].

[0141] Treatment and Outcome Recombinant human pGSN (rhu-pGSN) was synthesized in Escherichia coli and purified by Fujifilm Diosynth (Billingham, UK). Based on previous demonstrations of the function of rhu-pGSN in rodent models and data using human gelsolin to facilitate clinical translation efforts, human gelsolin rather than mouse gelsolin was used. rhu-pGSN was administered daily to mice by subcutaneous injection starting on day 3 or 6 post-infection at doses in the range of 0.5 - 5 mg, as detailed in the results. Mice were monitored for 12 days, and composite indices (i.e., rounded back appearance, ruffled fur, or partially closed eyes each worth 1 point; penile protrusion or splayed hindquarters worth 1.5 points; lethargy worth 2 points; maximum score is 8; evaluation was performed without blinding the treatment groups) adapted from previously described guidelines [Burkholder T et al., Current Protocols Mouse Biol. 2012;2: 145 - 65.] were used to measure survival, weight change, and overall morbidity. For animals that did not survive, the weight and morbidity score on the last day of survival were carried forward.

[0142] Lung transcriptome profiling Lung tissues were harvested from mice treated with either vehicle or rhu-pGSN on days 7 and 9 post-infection (2 mg per day was administered starting on day 3 post-infection and then increased to 5 mg per day on day 7). RNA was isolated using the RNAEasy Mini Kit (Qiagen, Germantown, MD) according to the manufacturer's instructions. RNA samples were analyzed using the Mouse DriverMap Target Gene Expression Profiling Panel from Cellecta (Mountain View, CA). The Cellecta platform measures the expression of 4753 functionally important mouse genes that encode proteins using highly multiplexed RT-PCR amplification and next-generation sequencing (NGS) quantification. Amplified indexed libraries sequenced on an Illumina NextSeq 500 instrument were created according to the procedures detailed in item 5.3 of the Cellecta User Manual. Sequencing data were converted to FASTQ format and further analyzed using DriverMap Sample Extraction software. This generated a raw data matrix file of counts for each sample for the columns aligned to the 4753 gene panel.

[0143] Statistical analysis Data were analyzed using Prism (GraphPad Software) or SAS (SAS Institute) software. Differences in Kaplan–Meier survival curves were analyzed using the log-rank test with Sidak adjustment for multiple comparisons. The Breslow–Day test for homogeneity of the pGSN versus vehicle comparison across the study gave p > 0.2, indicating that homogeneity was not rejected and supporting the overall study comparison performed via the stratified log-rank (Mantel–Cox) test by trial. For other measurements, differences between groups were investigated by ANOVA. Results of transcriptome profiling, scaled to normalize column counts, were transformed to log2 counts (after adding 0.1 to all cells to remove zero values) and analyzed using Qlucore software (Lund, Sweden). Further analysis of gene set enrichment was performed using tools (Panther version 14.118 and MetaCore (version 19.3, Clarivate Analytics, Philadelphia, PA)) that enabled assessment using a custom background gene list (i.e., approximately 4700 genes measured using the Cellecta DriverMap platform).

[0144] Results Effect of rhu-pGSN on survival rate To evaluate the potential of rhu-pGSN to improve outcomes, various dose and timing regimens were tested, and a total of 18 tests were conducted, summarized in a table in Figure 12 and summarized in Figure 13. To mimic possible clinical usage, mice were not treated until several days after challenge.

[0145] The main finding was that the delayed treatment with rhu-pGSN resulted in a significant improvement in the survival rate of mice (Figs. 14A-H). Combining all the studies, 39% (93 / 236) of the surviving mice treated with vehicle and 62% (241 / 389) of the surviving mice treated with pGSN were obtained on day 12 (p = 0.000001, Fig. 14A). An improvement in the survival rate was observed regardless of whether the delayed treatment was initiated on day 6 (Fig. 14C) or day 3 (Figs. 14E, 14G) post-infection. Similarly, the morbidity score was reduced by rhu-pGSN compared to vehicle treatment (Figs. 14B, 14D, 14F, 14H). In contrast, no statistically significant differences in weight loss or recovery (in surviving animals) were consistently observed in the experiments summarized in Figs. 14A-H. The only exception was found in a study that initially tested a low-dose regimen (>2 mg of rhu-pGSN on days 3-6 / 7, then 5 mg until day 11). In this latter series of studies, the body weight at the end of the study was 81.4 ± 4.7% in vehicle-treated mice compared to 85 ± 2.6% in pGSN-treated mice (p < 0.0001, summary of 4 studies, and also Figs. 12 and 13, and more detailed tables of all experiments in the Extended Data [Kobzik L: "Expanded Tables 1 & 2". Harvard Dataverse, V1 2019. www.doi.org / 10.7910 / DVN / 53GJY1]). The beneficial effects of rhu-pGSN were observed in most, but not all, of the 18 individual studies (see Fig. 12 and discussion).

[0146] Transcriptome profiling To assess whether rhu-pGSN treatment altered the transcriptome profile of infected lungs (see Harvard Dataverse: Expanded Tables 1 & 2. / / doi.org / 10.7910 / DVN / 53GJY116), lung tissues were harvested immediately before (day 7) and after (day 9) the onset of normal death (day 8) of this model (n = 5 per group per day). For each protocol, the dose of rhu-pGSN increased on day 7 of this experiment between the two time points selected for profiling. Comparison of lung samples obtained on day 7 from vehicle-treated and rhu-pGSN-treated mice showed no significant differences. In contrast, analysis of day 9 samples identified 344 differentially expressed genes in the rhu-pGSN-treated group, consisting of 195 downregulated genes and 149 upregulated genes. The top 50 upregulated and downregulated genes are shown in Figure 15, which is notable for the large number of cytokines and immune-related genes among the downregulated genes (especially IL10, IL12rb, CTLA4, and CCR9, 7, and 5 etc.) in the rhu-pGSN-treated group. Gene enrichment analysis of the complete downregulated gene list was performed using the Panther online analysis tool, querying the GO ontology or Reactome database. The main findings were a decrease in the expression of biological processes related to immune and inflammatory responses, or the release of cytokines and other cell activators. The top 10 most important processes / pathways are shown in Figure 16. Analysis using another gene enrichment analysis software tool (MetaCore) also produced similar results. Analysis of the upregulated gene list identified enrichment of processes related to tissue morphogenesis and epithelial / epidermal cell differentiation (see discussion, consistent with repair of influenza-mediated damage).The full results of gene enrichment analysis using the DriverMap gene list details, the identified differentially expressed genes, and the downregulation and upregulation gene lists to query the Panther and MetaCore databases are presented in worksheets 2 - 15 in the spreadsheets available in the Extended Data [Kobzik L: Harvard Dataverse, V1 2019. www.doi.org / 10.7910 / DVN / 8HBFD7]. Data regarding the experimental groups of the study described herein are shown in Tables 1 and 2 of Harvard Dataverse: Expanded Tables 1 & 2. / / doi.org / 10.7910 / DVN / 53GJY116, which also describes additional variables such as weights and statistical analysis. Additional data from the experiments described herein are provided by NCBI Gene Expression Omnibus: Transcriptome profiling of lung tissue from influenza-infected mice treated with plasma gelsolin. Accession number GSE138986; / / identifiers.org / geo:GSE138986.

[0147] Discussion A study was performed to evaluate the potential of rhu-pGSN to improve the outcome of severe influenza using a clinically relevant scenario of delaying treatment initiation. The key finding was that delaying pGSN treatment significantly improved survival, whether used starting on day 3 or day 6 post-infection. Delays were implemented to avoid interfering with the immediate immune response to influenza, given that it may not be possible to initiate early treatment immediately after patient infection (as opposed to immediately upon the onset of severe symptoms), and considering the adverse outcomes observed in some experimental models.

[0148] Some limitations are worthy of consideration. First, there is the observed experimental variability. Treatment with rhu-pGSN increased survival in most of the experiments conducted, but not in all of them. In some negative tests, factors such as technical problems with the virus stock, variations in the drop infusion method, and insufficient initial rhu-pGSN dosage in the "low dose, then high dose" tests, among others, but not limited to these, were considered to be the result. As much as possible, the methods were adjusted to reduce these potential sources of variability.

[0149] The experimental variables were also manipulated to investigate, for example, whether a late treatment on day 6 versus day 3 after the start of infection was effective, and other variables of the study were evaluated. Ultimately, a beneficial effect was observed regardless of whether all the tests were included in the survival analysis (Figures 14A, B), or whether the tests using treatment starting on day 6 or day 3 were included (Figures 14C-H).

[0150] When euthanasia was performed on surviving mice, the mice were followed for only 12 days. Since the survival curve was still potentially declining, the final mortality could not be confirmed with certainty. However, when rhu-pGSN was used compared to placebo treatment, the period until death was at least extended.

[0151] In particular, rhu-pGSN did not rescue all the mice that died from influenza in the experimental model, but the results showed a significant survival prolongation effect. Considering the goal of identifying a new treatment for severe influenza, the results obtained in mice without supportive fluids, additional therapeutic agents (not limited to antiviral agents, for example), and respiratory care given to hospitalized patients support the conclusion that the method will bring similar benefits and synergistic benefits in the clinical setting. The results suggest that administration of the gelseptin agent at a suitable time after infection in combination with standard therapies such as antiviral drugs provides a greater survival prolongation effect.

[0152] In summary, rhu-pGSN can improve the outcomes of a highly lethal mouse influenza model when administered after a clinically appropriate delay. These findings are consistent with the benefits seen in models of pneumococcal pneumonia. The mechanism of action of pGSN appears to involve the host response and is not dependent on a specific type of pathogen. The experimental results support the use of gelsolin as an adjuvant therapy for severe influenza and other viral infections in humans and other mammals.

[0153] Example 5 Additional studies are performed using a synergistic amount of a gelsolin agent and an antiviral agent. In certain studies, oseltamivir phosphate, zanamivir, peramivir, or baloxavir marboxil is the antiviral agent administered to the subject. The gelsolin agent is administered by a delayed dosing regimen as described above herein. An effective amount of the antiviral agent and the gelsolin agent are administered to a subject having or suspected of having a viral infection such as one of influenza A, B, C, or D, and the effective amount results in a synergistic therapeutic effect against the virus in the subject's infection. The synergistic therapeutic effect more greatly improves one or more characteristics of the viral infection in the subject than improvement of one or more characteristics in a control, where the control has not received a treatment comprising administration of a synergistic effective amount of the gelsolin agent and the antiviral agent.

[0154] Equivalents Although several embodiments of the present invention have been described and explained herein, those skilled in the art can readily envision various other means and / or structures for performing the functions described herein and / or obtaining the results and / or one or more advantages, and each such variation and / or modification is considered to be within the scope of the present invention. More generally, those skilled in the art will recognize that all parameters, dimensions, materials, and configurations described herein are intended to be exemplary, and that actual parameters, dimensions, materials, and / or configurations will depend on the particular application or applications for which the teachings of the present invention are used. Those skilled in the art will be able to recognize, or confirm with only routine experimentation, many equivalents to the specific embodiments of the invention described herein. Accordingly, the foregoing embodiments are presented by way of example only, and it should be understood that within the scope of the appended claims and their equivalents, embodiments of the present invention may be practiced otherwise than as specifically described and claimed. The present invention relates to the individual features, systems, articles, materials, and / or methods described herein. Further, any combination of two or more of such features, systems, articles, and / or methods is included within the scope of the present invention if such features, systems, articles, and / or methods are not mutually inconsistent.

[0155] All definitions, as defined and used herein, are to be understood to use a distinction between the dictionary definition, the definition in the document incorporated by reference, and / or the ordinary meaning of the defined term.

[0156] As used in this specification and the appended claims, the indefinite articles "a" and "an" are to be understood to mean "at least one" unless clearly indicated to the contrary. As used in this specification and the claims, the phrase "and / or" is to be understood to mean "one or both" of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Whether or not specifically identified with another element, other elements may optionally be present in addition to those specifically identified by the "and / or" clause, unless there is a clear contrary indication.

[0157] All references, patents, patent applications, and publications cited or referenced in this application are hereby incorporated by reference in their entirety. Aspects of the Invention [Aspect 1] A composition comprising an effective amount of a gelseculin agent and an antibacterial agent for synergistically treating a microbial infection in a subject. [Aspect 2] The composition according to Aspect 1, wherein the antibacterial agent is in a clinically acceptable amount, and the administered gelseculin agent and antibacterial agent synergistically enhance the therapeutic effect of administering a clinically acceptable amount of the antibacterial agent to the subject without administering the gelseculin agent. [Aspect 3] The composition according to Aspect 1, wherein the clinically acceptable amount of the antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent in the subject. [Aspect 4] The composition according to Aspect 1, wherein the MTD of the antibacterial agent is the highest possible but still acceptable dose level of the antibacterial agent for the subject. [Aspect 5] The composition according to Aspect 4, wherein the MTD of the antibacterial agent is determined based at least in part on a clinically selected pre-determined limit toxicity for the antibacterial agent in the subject. [Aspect 6] The composition according to Aspect 1, wherein the synergistic effective amount of the gelseculin agent and the antibacterial agent reduces the minimum effective dose (MED) of the antibacterial agent in the subject. [Aspect 7] The composition according to Aspect 6, wherein the MED is the lowest dose level of the antibacterial agent that provides a clinically significant response in average effectiveness, and the response is statistically significantly greater than the response provided by a control that does not include the dose of the antibacterial agent. [Aspect 8] The composition according to Aspect 1, wherein the synergistic therapeutic effect of the gelseculin agent and the antibacterial agent includes enhancing the viability of the subject. [Aspect 9] The composition according to aspect 1, comprising reducing microbial infection in a subject by the synergistic therapeutic effect of a gelsonlin agent and an antibacterial agent. [Aspect 10] The composition according to aspect 1, wherein the microbial infection is a bacterial infection, optionally caused by a pneumococcal species. [Aspect 11] The composition according to aspect 1, wherein the antibacterial agent comprises a β-lactam antibiotic. [Aspect 12] The composition according to aspect 1, wherein the antibacterial agent comprises penicillin. [Aspect 13] The composition according to aspect 1, wherein the microbial infection is caused by one type of Pseudomonas aeruginosa. [Aspect 14] The composition according to aspect 1, wherein the antibacterial agent is a carbapenem-class antibacterial agent. [Aspect 15] The composition according to aspect 1, wherein the antibacterial agent is meropenem. [Aspect 16] The composition according to aspect 1, wherein the antibacterial agent comprises an antifungal agent and the microbial infection comprises a fungal infection. [Aspect 17] The composition according to aspect 1, wherein the antibacterial agent comprises an antiparasitic agent and the microbial infection comprises a parasitic infection. [Aspect 18] The composition according to aspect 1, wherein the antibacterial agent comprises an antiviral agent and the microbial infection comprises a viral infection. [Aspect 19] The composition according to aspect 1, wherein the subject is a mammal, optionally a human. [Aspect 20] The composition according to aspect 1, wherein the gelsonlin agent comprises plasma gelsonlin (pGSN), optionally recombinant pGSN. [Aspect 21] The composition according to aspect 1, further comprising a pharmaceutically acceptable carrier. [Aspect 22] The composition according to aspect 1, wherein the gelsonlin agent comprises a gelsonlin molecule, a functional fragment thereof, or a functional derivative of the gelsonlin molecule. [Aspect 23] The composition according to aspect 1, further comprising a pharmaceutically acceptable carrier. [Aspect 24] A method of increasing the therapeutic effect of an antibacterial agent against a microbial infection in a subject, comprising: A method comprising the step of administering to a subject having a microbial infection a synergistically effective amount of each of a gelsolin agent and an antibacterial agent, wherein the administered gelsolin agent and antibacterial agent have a synergistic therapeutic effect against the microbial infection of the subject, and the synergistic therapeutic effect is greater than the therapeutic effect of the antibacterial agent administered without the gelsolin agent. [Aspect 25] The method according to aspect 24, wherein the antibacterial agent is administered in a clinically acceptable amount. [Aspect 26] The method according to aspect 24, wherein the synergistic therapeutic effect against the microbial infection is higher than the control therapeutic effect against the microbial infection, and the control therapeutic effect is the sum of the therapeutic effect of the antibacterial agent against the microbial infection and the therapeutic effect of the gelsolin agent against the microbial infection when each of the antibacterial agent and the gelsolin agent is administered without the other. [Aspect 27] The method according to aspect 26, wherein the control therapeutic effect is equal to the individual therapeutic effect of the gelsolin agent. [Aspect 28] The method according to aspect 26, wherein the control therapeutic effect is equal to the individual therapeutic effect of the antibacterial agent administered in a clinically acceptable amount. [Aspect 29] The method according to aspect 27, wherein the synergistic therapeutic effect is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the control therapeutic effect. [Aspect 30] The method according to aspect 24, wherein the antibacterial agent comprises an antibiotic preparation and the microbial infection comprises a bacterial infection. [Aspect 31] The method according to aspect 24, wherein the antibacterial agent comprises an antifungal agent and the microbial infection comprises a fungal infection. [Aspect 32] The method according to aspect 24, wherein the antibacterial agent comprises an antiparasitic agent and the microbial infection comprises a parasitic infection. [Aspect 33] The method according to aspect 24, wherein the antibacterial agent comprises an antiviral agent and the microbial infection comprises a viral infection. [Aspect 34] The method according to aspect 24, wherein the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of the gelsolin molecule. [Aspect 35] The method according to aspect 34, wherein the gelsolin molecule is plasma gelsolin (pGSN). [Aspect 36] The method according to aspect 34 or 35, wherein the gelsolin molecule is a recombinant gelsolin molecule. [Aspect 37] The method according to aspect 25, wherein the clinically acceptable amount of the antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent. [Aspect 38] The method according to aspect 37, wherein the MTD of the antibacterial agent is the highest possible but still acceptable dose level of the antibacterial agent for the subject. [Aspect 39] The method according to aspect 38, wherein the MTD of the antibacterial agent is determined based at least in part on a clinically selected pre-determined limit toxicity for the antibacterial agent. [Aspect 40] The method according to aspect 24, wherein the synergistically effective amount of the gelsolin agent and the antibacterial agent reduces the minimum effective dose (MED) of the antibacterial agent in the subject. [Aspect 41] The method according to aspect 24, wherein the synergistic therapeutic effect of the administration of each of the synergistically effective amount of the antibacterial agent and the gelsolin agent reduces the level of microbial infection in the subject as compared to the level of control microbial infection. [Aspect 42] The method according to aspect 41, wherein the control infection level includes the infection level when each of the synergistically effective amount of the antibacterial agent and the gelsolin agent is not administered. [Aspect 43] The method according to aspect 41 or 42, wherein the level of microbial infection in the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% lower than the level of control microbial infection. [Aspect 44] The method according to aspect 41, wherein the level of microbial infection in the subject is determined, and the means of determination includes one or more of an assay, observing the subject, evaluating one or more physiological symptoms of microbial infection in the subject, and evaluating the viability of the subject. [Aspect 45] The method according to aspect 44, wherein the physiological symptoms include one or more of fever, malaise, and death. [Aspect 46] The method according to aspect 44, wherein the physiological symptoms include lung lesions. [Aspect 47] The method according to aspect 44, wherein the physiological symptom includes weight loss. [Aspect 48] The method according to aspect 44, wherein the assay includes means for detecting the presence, absence, and / or level of characteristics of a microbial infection in a biological sample from the subject. [Aspect 49] The method according to aspect 24, wherein the administration of a synergistically effective amount of each of the antibacterial agent and the gelsecide increases the viability of the subject as compared to the viability of a control. [Aspect 50] The method according to aspect 49, wherein the viability of the control is the viability in the absence of the administration of a synergistically effective amount of each of the antibacterial agent and the gelsecide. [Aspect 51] The method according to aspect 49 or 50, wherein the increase in the viability of the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the viability of the control. [Aspect 52] The method according to aspect 24, wherein the administration of a synergistically effective amount of each of the antibacterial agent and the gelsecide reduces the level of lung lesions in the subject as compared to the level of lung lesions in a control. [Aspect 53] The method according to aspect 52, wherein the level of lung lesions in the control is the level of lung lesions in the absence of the administration of a synergistically effective amount of each of the antibacterial agent and the gelsecide. [Aspect 54] The method according to aspect 52 or 53, wherein the level of lung lesions in the subject administered a synergistically effective amount of each of the antibacterial agent and the gelsecide is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% lower than the level of lung lesions in the control. [Aspect 55] The method according to aspect 52, wherein the subject has a Pseudomonas aeruginosa bacterial infection. [Aspect 56] The method according to aspect 52, wherein the antibacterial agent includes the carbapenem class and optionally includes meropenem. [Aspect 57] The method according to aspect 30, wherein the bacterial infection is caused by one type of Streptococcus pneumoniae. [Aspect 58] The method according to aspect 30, wherein the antibacterial agent comprises a β-lactam antibiotic. [Aspect 59] The method according to aspect 30, wherein the antibacterial agent comprises penicillin. [Aspect 60] The method according to aspect 30, wherein the bacterial infection is caused by one type of Pseudomonas aeruginosa. [Aspect 61] The method according to aspect 60, wherein the antibacterial agent is a carbapenem-class antibacterial agent. [Aspect 62] The method according to aspect 61, wherein the antibacterial agent is meropenem. [Aspect 63] The method according to aspect 30, wherein the bacterial infection is caused by one or more of gram-positive bacteria, gram-negative bacteria, Mycobacterium tuberculosis, non-tuberculous mycobacteria, spirochetes, actinomycetes, Ureaplasma species bacteria, Mycoplasma species bacteria, and Chlamydia species bacteria. [Aspect 64] The method according to aspect 24, wherein the administration means of the gelseptin agent and the antibacterial agent are independently selected from oral, sublingual, buccal, intranasal, intravenous, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intra-synovial, and intraocular administrations. [Aspect 65] The method according to aspect 24, wherein the subject is a mammal, optionally a human. [Aspect 66] The method according to aspect 24, wherein the gelseptin agent is a non-therapeutic gelseptin agent. [Aspect 67] The method according to aspect 24, wherein the antibacterial agent is a non-therapeutic agent. [Aspect 68] A method for synergistically treating a microbial infection in a subject, comprising the step of administering to a subject having a microbial infection an effective amount of each of a gelseptin agent and an antibacterial agent, wherein the administered gelseptin agent and antibacterial agent have a synergistic therapeutic effect on the microbial infection of the subject as compared to the therapeutic effect of a control, and the antibacterial agent is administered in a clinically acceptable amount. [Aspect 69] The method according to aspect 68, wherein the control comprises the therapeutic effect of administering a clinically acceptable amount of an antibacterial agent administered without administering a gelseptin agent. [Aspect 70] The method according to aspect 68, wherein the clinically acceptable amount of the antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent. [Aspect 71] The method according to aspect 70, wherein the MTD of the antibacterial agent is the highest possible but still acceptable dose level of the antibacterial agent for the subject. [Aspect 72] The method according to aspect 71, wherein the MTD of the antibacterial agent is determined based at least in part on a clinically selected pre - determined limit toxicity for the antibacterial agent. [Aspect 73] The method according to aspect 68, wherein the synergistic effective amount of the gelseolin agent and the antibacterial agent reduces the minimum effective dose (MED) of the antibacterial agent in the subject. [Aspect 74] The method according to aspect 73, wherein the MED is the lowest dose level of the antibacterial agent that provides a clinically significant response in average effectiveness, and the response is statistically significantly greater than the response provided by a control that does not include the dose of the antibacterial agent. [Aspect 75] The method according to aspect 68, wherein the synergistic therapeutic effect is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the therapeutic effect of the control. [Aspect 76] The method according to aspect 68, wherein the antibacterial agent includes an antibiotic preparation and the microbial infection includes a bacterial infection. [Aspect 77] The method according to aspect 68, wherein the antibacterial agent includes an antifungal agent and the microbial infection includes a fungal infection. [Aspect 78] The method according to aspect 68, wherein the antibacterial agent includes an antiparasitic agent and the microbial infection includes a parasitic infection. [Aspect 79] The method according to aspect 68, wherein the antibacterial agent includes an antiviral agent and the microbial infection includes a viral infection. [Aspect 80] The method according to aspect 68, wherein the gelseolin agent includes a gelseolin molecule, a functional fragment thereof, or a functional derivative of the gelseolin molecule. [Aspect 81] The method according to aspect 68, wherein the gelseolin molecule is plasma gelseolin (pGSN). [Aspect 82] The method according to aspect 80 or 81, wherein the gelsolin molecule is a recombinant gelsolin molecule. [Aspect 83] The method according to aspect 68, wherein the synergistic therapeutic effect of the administration of each of the synergistically effective amounts of the antibacterial agent and the gelsolin agent reduces the level of microbial infection in the subject as compared to the level of control microbial infection. [Aspect 84] The method according to aspect 83, wherein the control infection level includes the infection level when each of the synergistically effective amounts of the antibacterial agent and the gelsolin agent is not administered. [Aspect 85] The method according to aspect 83, wherein the level of microbial infection in the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% lower than the level of control microbial infection. [Aspect 86] The method according to aspect 83, wherein the level of microbial infection in the subject is determined, and the means of determination includes one or more of an assay, observing the subject, evaluating one or more physiological symptoms of microbial infection in the subject, and evaluating the viability of the subject. [Aspect 87] The method according to aspect 86, wherein the physiological symptoms include one or more of fever, malaise, and death. [Aspect 88] The method according to aspect 86, wherein the physiological symptoms include weight loss. [Aspect 89] The method according to aspect 86, wherein the physiological symptoms include lung lesions. [Aspect 90] The method according to aspect 86, wherein the assay includes means for detecting the presence, absence, and / or level of characteristics of microbial infection in a biological sample from the subject. [Aspect 91] The method according to aspect 68, wherein the administration of each of the synergistically effective amounts of the antibacterial agent and the gelsolin agent increases the viability of the subject as compared to the viability of the control. [Aspect 92] The method according to aspect 91, wherein the control viability is the viability when each of the synergistically effective amounts of the antibacterial agent and the gelsolin agent is not administered. [Aspect 93] The method according to Aspect 91, wherein the increase in the viability of the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the viability of the control. [Aspect 94] The method according to Aspect 68, wherein the administration of each of the synergistically effective amounts of the antibacterial agent and the gelsecrin agent reduces the level of lung lesions in the subject as compared to the level of lung lesions in the control. [Aspect 95] The method according to Aspect 94, wherein the level of lung lesions in the control is the level of lung lesions in the absence of the administration of each of the synergistically effective amounts of the antibacterial agent and the gelsecrin agent. [Aspect 96] The method according to Aspect 94, wherein the level of lung lesions in the subject administered each of the synergistically effective amounts of the antibacterial agent and the gelsecrin agent is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% lower than the level of lung lesions in the control. [Aspect 97] The method according to Aspect 76, wherein the subject has a Pseudomonas aeruginosa bacterial infection. [Aspect 98] The method according to Aspect 94, wherein the antibacterial agent includes the carbapenem class and optionally includes meropenem. [Aspect 99] The method according to Aspect 76, wherein the bacterial infection is caused by one type of Streptococcus pneumoniae. [Aspect 100] The method according to Aspect 76, wherein the antibacterial agent includes a β-lactam antibiotic. [Aspect 101] The method according to Aspect 76, wherein the antibacterial agent includes penicillin. [Aspect 102] The method according to Aspect 76, wherein the bacterial infection is caused by one or more of gram-positive bacteria, gram-negative bacteria, Mycobacterium tuberculosis, nontuberculous mycobacteria, spirochetes, actinomycetes, Ureaplasma species bacteria, Mycoplasma species bacteria, and Chlamydia species bacteria. [Aspect 103] The method according to aspect 68, wherein the administration means of the gelsonlin agent and the antibacterial agent are independently selected from oral, sublingual, buccal, intranasal, intravenous, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intra-synovial sac, and intraocular administration. [Aspect 104] The method according to aspect 68, wherein the subject is a mammal. [Aspect 105] The method according to aspect 68, wherein the gelsonlin agent is a non-therapeutic gelsonlin agent. [Aspect 106] The method according to aspect 68, wherein the antibacterial agent is a non-therapeutic agent. [Aspect 107] A pharmaceutical composition comprising an antibacterial agent and a gelsonlin agent for use in a method of treating a subject, which synergistically increases the therapeutic effect of the antibacterial agent against a microbial infection, wherein the subject has a microbial infection, and the method comprises administering to the subject a pharmaceutical composition comprising a synergistically effective amount of each of the gelsonlin agent and the antibacterial agent in an amount effective to treat the microbial infection in the subject, and the synergistic therapeutic effect is greater than the therapeutic effect of the antibacterial agent administered without the gelsonlin agent. [Aspect 108] The pharmaceutical composition according to aspect 107, wherein the gelsonlin agent and the antibacterial agent are administered to the subject separately or simultaneously. [Aspect 109] The pharmaceutical composition according to aspect 107, wherein the antibacterial agent is administered in a clinically acceptable amount, and the administered gelsonlin agent and antibacterial agent synergistically enhance the therapeutic effect of administering a clinically acceptable amount of the antibacterial agent to the subject without administering the gelsonlin agent. [Aspect 110] The pharmaceutical composition according to aspect 109, wherein the clinically acceptable amount of the antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent in the subject. [Aspect 111] The pharmaceutical composition according to aspect 110, wherein the MTD of the antibacterial agent is the highest possible but still acceptable dose level of the antibacterial agent for the subject. [Aspect 112] The pharmaceutical composition according to aspect 110, wherein the MTD of the antibacterial agent is determined based at least in part on a clinically selected pre-determined limit toxicity for the antibacterial agent in the subject. [Aspect 113] The pharmaceutical composition according to aspect 107, wherein a synergistically effective amount of a gelsonlin agent and an antibacterial agent reduces the minimum effective dose (MED) of the antibacterial agent in a subject. [Aspect 114] The pharmaceutical composition according to aspect 113, wherein the MED is the lowest dose level of an antibacterial agent that provides a clinically significant response in mean effectiveness, and the response is statistically significantly greater than the response provided by a control that does not include the dose of the antibacterial agent. [Aspect 115] The pharmaceutical composition according to aspect 107, wherein the synergistic therapeutic effect of the gelsonlin agent and the antibacterial agent includes enhancing the viability of the subject. [Aspect 116] The pharmaceutical composition according to aspect 107, wherein the synergistic therapeutic effect of the gelsonlin agent and the antibacterial agent includes reducing a microbial infection in the subject. [Aspect 117] The pharmaceutical composition according to aspect 107, wherein the microbial infection is a bacterial infection, optionally caused by a pneumococcal species. [Aspect 118] The pharmaceutical composition according to aspect 107, wherein the antibacterial agent includes penicillin. [Aspect 119] The pharmaceutical composition according to aspect 107, wherein the bacterial infection is caused by one type of Pseudomonas aeruginosa. [Aspect 120] The pharmaceutical composition according to aspect 107, wherein the antibacterial agent is a carbapenem class antibacterial agent. [Aspect 121] The pharmaceutical composition according to aspect 107, wherein the antibacterial agent is meropenem. [Aspect 122] The pharmaceutical composition according to aspect 107, wherein the antibacterial agent includes an antifungal agent and the microbial infection includes a fungal infection. [Aspect 123] The pharmaceutical composition according to aspect 107, wherein the antibacterial agent includes an antiparasitic agent and the microbial infection includes a parasitic infection. [Aspect 124] The pharmaceutical composition according to aspect 107, wherein the antibacterial agent includes an antiviral agent and the microbial infection includes a viral infection. [Aspect 125] The pharmaceutical composition according to aspect 107, wherein the subject is a mammal. [Aspect 126] The pharmaceutical composition according to aspect 107, wherein the gelsonlin agent includes plasma gelsonlin (pGSN), and optionally, is recombinant pGSN. [Aspect 127] The pharmaceutical composition according to Aspect 107, further comprising a pharmaceutically acceptable carrier. [Aspect 128] The pharmaceutical composition according to Aspect 107, wherein the gelsevirin agent comprises a gelsevirin molecule, a functional fragment thereof, or a functional derivative of the gelsevirin molecule. [Aspect 129] The pharmaceutical composition according to Aspect 107, further comprising a pharmaceutically acceptable carrier. [Aspect 130] A method for treating viral infection in a subject, comprising the step of administering an effective amount of a gelsevirin agent to a subject having a viral infection, wherein the gelsevirin agent is administered at least 3, 4, 5, 6, 7, 8, 9 days, or more after the subject contracts the viral infection, and is not administered on the day the subject contracts the viral infection, 1 day after the subject contracts the viral infection, or 2 days after the subject contracts the viral infection. [Aspect 131] The method according to Aspect 130, wherein the effective amount of the gelsevirin agent increases the therapeutic effect on viral infection in the subject as compared to the therapeutic effect of a control. [Aspect 132] The method according to Aspect 131, wherein the therapeutic effect of the control includes the therapeutic effect when the gelsevirin agent is not administered to the subject. [Aspect 133] The method according to Aspect 130, wherein the antiviral agent comprises one or more of oseltamivir phosphate, zanamivir, peramivir, and baloxavir marboxil. [Aspect 134] The method according to Aspect 130, wherein the therapeutic effect of the administered gelsevirin agent is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the therapeutic effect of the control. [Aspect 135] The method according to Aspect 130, wherein the gelsevirin agent comprises a gelsevirin molecule, a functional fragment thereof, or a functional derivative of the gelsevirin molecule. [Aspect 136] The method according to Aspect 130, wherein the gelsevirin molecule is plasma gelsevirin (pGSN). [Aspect 137] The method according to aspect 135 or 136, wherein the gelsolin molecule is a recombinant gelsolin molecule. [Aspect 138] The method according to aspect 131, wherein the therapeutic effect of the administration of the gelsolin agent reduces the level of viral infection in the subject as compared to the level of the control viral infection, and the control infection level includes the infection level when the gelsolin agent is not administered. [Aspect 139] The method according to aspect 138, wherein the level of viral infection in the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% lower than the level of the control viral infection. [Aspect 140] The method according to aspect 139, wherein the level of viral infection in the subject is determined, and the means of determination includes one or more of an assay, observing the subject, evaluating one or more physiological symptoms of viral infection in the subject, and evaluating the viability of the subject. [Aspect 141] The method according to aspect 140, wherein the physiological symptoms include one or more of fever, malaise, weight loss, and death. [Aspect 142] The method according to aspect 140, wherein the assay includes means for detecting the presence, absence, and / or level of features of viral infection in a biological sample from the subject. [Aspect 143] The method according to aspect 140, wherein the administration of an effective amount of the gelsolin agent increases the viability of the subject as compared to the viability of the control. [Aspect 144] The method according to aspect 143, wherein the viability of the control is the viability in the absence of the administration of the gelsolin agent. [Aspect 145] The method according to aspect 143, wherein the increase in the viability of the subject is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, or 200% higher than the viability of the control. [Aspect 146] The method according to Aspect 130, wherein the administration means of the gelsolin agent is selected from oral, sublingual, buccal, intranasal, intravenous, inhalation, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intra-synovial sac, and intraocular administration. [Aspect 147] The method according to Aspect 130, wherein the subject is a mammal, optionally a human. [Aspect 148] The method according to any one of Aspects 130 to 147, further comprising the step of treating the subject with an antiviral agent more than one day before the administration of the gelsolin agent to the subject, and the antiviral agent is administered on one or more days out of the day when the subject contracts a viral infection, one day after the subject contracts a viral infection, and two days after the subject contracts a viral infection. [Aspect 149] The method according to Aspect 148, wherein a synergistically effective amount of each of the gelsolin agent and the antiviral agent is administered to the subject, and has a synergistic therapeutic effect against viral infection as compared to the therapeutic effect of a control, and the antiviral agent is administered in a clinically acceptable amount. [Aspect 150] The method according to Aspect 149, wherein the control includes the therapeutic effect of the administration of a clinically acceptable amount of an antiviral agent administered without administering the gelsolin agent. [Aspect 151] The method according to Aspect 150, wherein the clinically acceptable amount of the antiviral agent is an amount less than the maximum tolerated dose (MTD) of the antiviral agent. [Aspect 152] The method according to Aspect 151, wherein the MTD of the antiviral agent is the highest possible but still acceptable dose level of the antiviral agent for the subject. [Aspect 153] The method according to Aspect 152, wherein the MTD of the antiviral agent is determined based at least in part on a clinically selected pre-determined limit toxicity for the antiviral agent. [Aspect 154] The method according to Aspect 149, wherein the synergistically effective amount of the gelsolin agent and the antiviral agent reduces the minimum effective dose (MED) of the antiviral agent in the subject. [Aspect 155] The method according to Aspect 154, wherein the MED is the lowest dose level of the antiviral agent that provides a clinically significant response in terms of average effectiveness, and the response is statistically significantly greater than the response provided by a control not including the dose of the antiviral agent. [Aspect 156] The method according to Aspect 148, wherein the administration means of the gelseolin agent and the antiviral agent are independently selected from oral, sublingual, buccal, intranasal, inhalation, intravenous, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intra-synovial sac, and intraocular administrations.

Claims

**Claim 1** A composition comprising an effective amount of a gelsolin agent and an antibacterial agent for synergistically treating microbial infections in a subject, the composition comprises a clinically acceptable amount of the antibacterial agent, and the composition comprising the administered gelsolin agent and antibacterial agent synergistically enhances the therapeutic effect of administering a clinically acceptable amount of the antibacterial agent to the subject without administering the gelsolin agent, wherein (a) the antibacterial agent is penicillin and the microbial infection is a bacterial infection caused by penicillin-resistant pneumococcal species, or (b) the antibacterial agent is meropenem and the microbial infection is a bacterial infection caused by multi-drug resistant (MDR) Pseudomonas species. **Claim 2** The composition according to claim 1, wherein the clinically acceptable amount of the antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent in the subject, and optionally the MTD of the antibacterial agent is the highest possible but still acceptable dose level of the antibacterial agent for the subject. **Claim 3** The composition according to claim 2, wherein the MTD of the antibacterial agent is determined based at least in part on a clinically selected pre-determined limit toxicity for the antibacterial agent in the subject. **Claim 4** The composition according to claim 1, wherein the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of the gelsolin molecule, and optionally the gelsolin agent comprises plasma gelsolin (pGSN), and optionally the pGSN is recombinant pGSN. **Claim 5** A pharmaceutical composition comprising a gelsolin agent and an antibacterial agent for use in a method of increasing the therapeutic effect of an antibacterial agent against a microbial infection in a subject, the method comprising: administering to a subject having a microbial infection a synergistically effective amount of each of a gelsolin agent and an antibacterial agent, wherein the administered gelsolin agent and antibacterial agent have a synergistic therapeutic effect against the microbial infection of the subject, and the synergistic therapeutic effect is greater than the therapeutic effect of the antibacterial agent administered without the gelsolin agent, wherein (a) the antibacterial agent is penicillin and the microbial infection is a bacterial infection caused by penicillin-resistant pneumococcal species, or (b) the antibacterial agent is meropenem and the microbial infection is a bacterial infection caused by multi-drug resistant (MDR) Pseudomonas species. **Claim 6** The pharmaceutical composition according to claim 5, wherein the antibacterial agent is administered in a clinically acceptable amount, the clinically acceptable amount of the antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent, and the MTD of the antibacterial agent is determined based at least in part on a clinically selected limiting toxicity for the antibacterial agent.

7. The pharmaceutical composition according to claim 5, wherein the synergistic therapeutic effect against a microbial infection is higher than the control therapeutic effect against the microbial infection, and the control therapeutic effect is the sum of the therapeutic effect of the antibacterial agent against the microbial infection and the therapeutic effect of the gelsecrin agent against the microbial infection when each of the antibacterial agent and the gelsecrin agent is administered without the other.

8. The pharmaceutical composition according to claim 5, wherein the gelsecrin agent comprises a gelsecrin molecule, a functional fragment thereof, or a functional derivative of the gelsecrin molecule, optionally wherein the gelsecrin molecule is plasma gelsecrin (pGSN), and optionally wherein the gelsecrin molecule is a recombinant gelsecrin molecule.

9. The pharmaceutical composition according to claim 5, wherein the synergistic effective amounts of the gelsecrin agent and the antibacterial agent reduce the minimum effective dose (MED) of the antibacterial agent in the subject.

10. The pharmaceutical composition according to claim 5, wherein the administration of each of the synergistic effective amounts of the antibacterial agent and the gelsecrin agent increases the survival probability of the subject as compared to the control survival probability.

11. A pharmaceutical composition comprising a gelsecrin agent and an antibacterial agent for use in a method for synergistically treating a microbial infection in a subject, the method comprising administering to the subject having the microbial infection an effective amount of each of the gelsecrin agent and the antibacterial agent, wherein the administered gelsecrin agent and antibacterial agent have a synergistic therapeutic effect against the microbial infection of the subject as compared to the control therapeutic effect, and the antibacterial agent is administered in a clinically acceptable amount, (a) the antibacterial agent is penicillin and the microbial infection is a bacterial infection caused by a penicillin-resistant Streptococcus pneumoniae species, or (b) the antibacterial agent is meropenem and the microbial infection is a bacterial infection caused by a multi-drug resistant (MDR) Pseudomonas species.

12. The pharmaceutical composition according to claim 11, wherein the gelseolin agent comprises a gelseolin molecule, a functional fragment thereof, or a functional derivative of the gelseolin molecule, optionally the gelseolin molecule is plasma gelseolin (pGSN), and optionally the gelseolin molecule is a recombinant gelseolin molecule.

13. A pharmaceutical composition for use in a method of treating a subject, comprising an antibacterial agent and a gelseolin agent that synergistically increase the therapeutic effect of the antibacterial agent against a microbial infection, wherein the subject has a microbial infection, and the method comprises administering to the subject a pharmaceutical composition comprising a synergistically effective amount of each of the gelseolin agent and the antibacterial agent in an amount effective to treat the microbial infection in the subject, and the synergistic therapeutic effect is greater than the therapeutic effect of the antibacterial agent administered without the gelseolin agent. The pharmaceutical composition, wherein (a) the antibacterial agent is penicillin and the microbial infection is a bacterial infection caused by a penicillin-resistant Streptococcus pneumoniae species, or (b) the antibacterial agent is meropenem and the microbial infection is a bacterial infection caused by a multi-drug resistant (MDR) Pseudomonas species.

14. The pharmaceutical composition according to claim 13, wherein the gelseolin agent and the antibacterial agent are administered to the subject separately or simultaneously.

15. The pharmaceutical composition according to claim 13, wherein the antibacterial agent is administered in a clinically acceptable amount, and the administered gelseolin agent and antibacterial agent synergistically enhance the therapeutic effect of administering to the subject a clinically acceptable amount of the antibacterial agent without administering the gelseolin agent.

16. The pharmaceutical composition according to claim 15, wherein the clinically acceptable amount of the antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent in the subject, and optionally the MTD of the antibacterial agent is the highest possible but still acceptable dose level of the antibacterial agent for the subject.

17. The pharmaceutical composition according to claim 16, wherein the MTD of the antibacterial agent is determined based at least in part on a clinically selected pre-determined limit toxicity for the antibacterial agent in the subject.

18. The pharmaceutical composition according to claim 13, wherein the synergistically effective amount of the gelseolin agent and the antibacterial agent reduces the minimum effective dose (MED) of the antibacterial agent in the subject, and optionally the MED is the lowest dose level of the antibacterial agent that provides a clinically significant response in terms of average effectiveness, and the response is statistically significantly greater than the response provided by a control that does not include the dose of the antibacterial agent.

19. The pharmaceutical composition according to claim 13, wherein the gelsolin agent comprises plasma gelsolin (pGSN), optionally the pGSN is recombinant pGSN, and optionally the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of the gelsolin molecule.

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