Compositions and synergistic methods for treating infections

Abstract

To provide compositions that can be used to synergistically treat microbial infections.SOLUTION: A composition comprises effective amounts of a gelsolin agent and an antimicrobial agent for treating a microbial infection in a subject, where the composition comprises a clinically acceptable amount of the antimicrobial agent, and where the presence of the gelsolin agent in the composition reduces the minimum effective dose (MED) of the antimicrobial agent in the subject.SELECTED DRAWING: Figure 1A

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 June 21, 2019, the disclosure of which is incorporated herein 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] The present invention, in some embodiments, relates to compositions and methods for enhancing host immune defenses in the treatment of microbial infections. [Background technology]

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

[0004] Antibacterial agents have long been used to treat microbial infections because they have therapeutic effects against them in humans and animals. Resistance to previously therapeutically effective antibacterial agents can be caused by changes in the pathogens causing the infection. Overuse and misuse of antibacterial agents may be factors in the growing problem of antibacterial resistance, which has led to an ever-increasing number of types of pathogenic infections that are less responsive to previously effective antibacterial agents. Antibacterial resistance results in a lack of therapeutic options for treating pathogenic infections. Antibacterial-resistant pathogens cause many deaths each year and are a serious public health challenge worldwide. Summary of the Invention [Means for solving the problem]

[0005] The present invention relates, in part, to compositions that can be used to synergistically treat microbial infections. The compositions include one or more antibacterial agents and a gelsolin agent. The methods of the present invention relate, in part, to administering such compositions to a subject, wherein the antibacterial agent and gelsolin are administered synergistically. The Luzoline agents act synergistically to treat a microbial infection in a subject.

[0006] According to one aspect of the present invention, a composition is provided comprising an effective amount of a gelsolin agent and an antibacterial agent for synergistically treating a microbial infection in a subject. In some embodiments, the antibacterial agent is in a clinically tolerated amount, and the administered gelsolin agent and antibacterial agent synergistically enhance the therapeutic effect of administering a clinically tolerated amount of the antibacterial agent to a subject without administering the gelsolin agent. In certain embodiments, the clinically tolerated 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, yet tolerable, dose level of the antibacterial agent in the subject. In some embodiments, the MTD of the antibacterial agent is determined at least in part based on a preselected clinically limiting toxicity of the antibacterial agent in the subject. In certain embodiments, 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. In certain embodiments, the MED is the lowest dose level of the antibacterial agent that provides a clinically significant response in mean efficacy, which response is statistically significantly greater than the response provided by a control without that dose of the antibacterial agent. In some embodiments, the synergistic therapeutic effect of the gelsolin agent and the antibacterial agent comprises increasing the survival chances of the subject. In some embodiments, the synergistic therapeutic effect of the gelsolin agent and the antibacterial agent comprises reducing a microbial infection in the subject. In some embodiments, the microbial infection is a bacterial infection, optionally caused by a Streptococcus pneumoniae species. In certain embodiments, the antibacterial agent comprises a β-lactam antibiotic. In some embodiments, the antibacterial agent comprises a penicillin. In some embodiments, the microbial infection is caused by a 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 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 certain embodiments, the antibacterial agent comprises an antiviral agent, and the microbial infection comprises 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 a 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 is provided, comprising administering to a subject having a microbial infection synergistically effective amounts 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, the synergistic therapeutic effect being 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 greater than a control therapeutic effect against the microbial infection, the control therapeutic effect being the sum of the therapeutic effect of the antibacterial agent against the microbial infection when each of the antibacterial agent and the gelsolin agent is administered without the other and the therapeutic effect of the gelsolin agent against the microbial infection. 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% greater than the therapeutic effect of a control. In some embodiments, the antimicrobial agent comprises an antibiotic agent and the microbial infection comprises a bacterial infection. In some embodiments, the antimicrobial agent comprises an antifungal agent and the microbial infection comprises a microorganism. The biological 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 a 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, a clinically tolerated amount of an antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent. In some embodiments, the MTD of an antibacterial agent is the highest possible, yet tolerable, dose level of the antibacterial agent for a subject. In certain embodiments, the MTD of an antibacterial agent is determined at least in part based on a preselected clinically limiting toxicity for the antibacterial agent. In some embodiments, a synergistically effective amount of a gelsolin agent and an antibacterial agent reduces the minimum effective dose (MED) of the antibacterial agent in a subject. In some embodiments, the synergistic therapeutic effect of administering a synergistically effective amount of each of the antimicrobial agent and the gelsolin agent reduces the level of microbial infection in the subject compared to the level of microbial infection in a control. In some embodiments, the control level of infection comprises the level of infection in the absence of the synergistically effective amounts of each of the antimicrobial agent and the 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 microbial infection in the control. In some embodiments, the level of microbial infection in the subject is determined, and the determining method includes one or more of an assay, observing the subject, assessing one or more physiological symptoms of 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 symptom comprises pulmonary pathology, hi some embodiments, the physiological symptom comprises weight loss.In some embodiments, the assay comprises a means for detecting the presence, absence, and / or level of a characteristic of a microbial infection in a biological sample from a subject. In some embodiments, administration of a synergistically effective amount of each of the antimicrobial agent and gelsolin increases the subject's viability compared to a control's viability. In certain embodiments, the control's viability is the viability in the absence of administration of the synergistically effective amounts of each of the antimicrobial agent and gelsolin. In some embodiments, the increase in the subject's viability 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% greater than the control's viability. In certain embodiments, administration of a synergistically effective amount of each of the antimicrobial agent and gelsolin reduces the level of lung lesions in the subject compared to the level of lung lesions in the control. In some embodiments, the control lung lesion level is the lung lesion level in the absence of administration of a synergistically effective amount of each of the antibacterial agent and gelsolin. In certain embodiments, the lung lesion level in a subject administered a synergistically effective amount of each of the antibacterial agent and gelsolin 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 control lung lesion level. In some embodiments, the subject has a Pseudomonas aeruginosa bacterial infection. In some embodiments, the antibacterial agent comprises a carbapenem class, optionally including meropenem. In certain embodiments, the bacterial infection is caused by a type of Streptococcus pneumoniae (pneumococcus). In some embodiments, the antibacterial agent comprises a β-lactam antibiotic. In some embodiments, the antibacterial agent comprises penicillin. In certain embodiments, the bacterial infection is caused by a 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. 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 some embodiments, the means of administration of the gelsolin agent and the antimicrobial agent are independently selected from oral, sublingual, buccal, intranasal, intravenous, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intrasynovial, 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 antimicrobial 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 a subject having a microbial infection effective amounts of a gelsolin agent and an antimicrobial agent, wherein the administered gelsolin agent and antimicrobial agent have a synergistic therapeutic effect on the microbial infection in the subject compared to the therapeutic effect of a control, wherein the antimicrobial agent is administered in a clinically tolerated amount. In some embodiments, the control comprises the therapeutic effect of administering a clinically tolerated amount of the antimicrobial agent without administering the gelsolin agent. In certain embodiments, the clinically tolerated amount of the antimicrobial agent is an amount less than the maximum tolerated dose (MTD) of the antimicrobial agent. In some embodiments, the MTD of the antimicrobial agent is the highest possible, yet tolerable, dose level of the antimicrobial agent in the subject. In some embodiments, the MTD of the antimicrobial agent is determined at least in part based on a preselected clinically limiting toxicity for the antimicrobial agent. In some embodiments, the synergistically effective amounts of the gelsolin agent and the antimicrobial agent reduce the minimum effective dose (MED) of the antimicrobial agent in the subject. In certain embodiments, the MED is the lowest dose level of an antimicrobial agent that provides a clinically significant response in mean efficacy, which response is statistically significantly greater than the response provided by a control without that dose of antimicrobial 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% greater than the therapeutic effect of the control. In some embodiments, the antimicrobial agent comprises an antibiotic agent, and the microbial infection comprises a bacterial infection. In certain embodiments, the antimicrobial 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 a 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 administering a synergistically effective amount of each of the antibacterial agent and the gelsolin agent reduces the level of microbial infection in the subject compared to a control level of microbial infection. In some embodiments, the control level of infection comprises the level of infection without administering a synergistically effective amount of each of the antibacterial agent and the 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 control level of microbial infection. In some embodiments, the level of microbial infection in the subject is determined, and the means for determining include one or more of assays, observing the subject, assessing one or more physiological symptoms of microbial infection in the subject, and assessing the viability of the subject. In certain embodiments, the physiological symptoms include one or more of fever, fatigue, and death. In some embodiments, the physiological symptoms include weight loss. In some embodiments, the physiological symptoms include pulmonary pathology. In certain embodiments, the assay detects the presence, absence, and / or level of a characteristic of a microbial infection in a biological sample from a subject. In some embodiments, administration of a synergistically effective amount of each of the antimicrobial agent and gelsolin agent increases the subject's viability compared to a control's viability. In certain embodiments, the control's viability is the viability in the absence of administration of a synergistically effective amount of each of the antimicrobial agent and gelsolin agent. In some embodiments, the increase in the subject's viability 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% greater than the control's viability. In some embodiments, administration of a synergistically effective amount of each of the gelsolin agent and the antimicrobial agent reduces the level of lung lesions in the subject compared to the level of lesions in the control. In certain embodiments, the control lung lesion level is the lung lesion level in the absence of administration of a synergistically effective amount of each of the antibacterial agent and gelsolin. In some embodiments, the lung lesion level in a subject administered a synergistically effective amount of each of the antibacterial agent and gelsolin 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 control lung lesion level. In some embodiments, the subject has a Pseudomonas aeruginosa bacterial infection. In certain embodiments, the antibacterial agent comprises a carbapenem class, optionally including meropenem. In some embodiments, the bacterial infection is caused by a type of Streptococcus pneumoniae (pneumococcus). In certain embodiments, the antibacterial agent comprises a β-lactam antibiotic. In some embodiments, the antibacterial agent comprises 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 gelsolin agent and the antimicrobial agent are independently selected from oral, sublingual, buccal, intranasal, intravenous, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intrasynovial, and intraocular administration. In some embodiments, the subject is a mammal. In some embodiments, the gelsolin agent is a non-therapeutic gelsolin agent. In certain embodiments, the antimicrobial agent is a non-therapeutic agent.

[0009] According to another aspect of the present invention, a pharmaceutical composition is provided for use in a method of treating a subject, the method comprising administering a pharmaceutical composition comprising a synergistically effective amount of each of the gelsolin and the antibacterial agent, the pharmaceutical composition comprising a synergistically effective amount of each of the gelsolin and the antibacterial agent, in an amount effective to treat the microbial infection in the subject, wherein the synergistic therapeutic effect is greater than the therapeutic effect of the antibacterial agent administered without the gelsolin. In some embodiments, the gelsolin and the antibacterial are administered separately or simultaneously to the subject. In certain embodiments, the antibacterial agent is administered in a clinically tolerated amount, and the administered gelsolin and antibacterial agents synergistically enhance the therapeutic effect of administering a clinically tolerated amount of the antibacterial agent to the subject without administering the gelsolin. In some embodiments, the clinically tolerated amount of the antibacterial agent is 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, yet tolerable, dose level of the antibacterial agent to the subject. In certain embodiments, the MTD of the antibacterial agent is determined at least in part based on a preselected clinical limiting toxicity for the antibacterial agent in the subject. In some embodiments, 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. In some embodiments, the MED is the lowest dose level of the antibacterial agent that provides a clinically significant response in mean efficacy, which response is statistically significantly greater than the response provided by a control that does not include that dose of the antibacterial agent. In certain embodiments, the synergistic therapeutic effect of the gelsolin agent and the antibacterial agent includes increasing the survival chances of the subject. In some embodiments, the synergistic therapeutic effect of the gelsolin agent and the antibacterial agent reduces microbial infection in the subject. In some embodiments, the microbial infection is a bacterial infection, optionally caused by a pneumococcus species. In certain embodiments, the antibacterial agent comprises penicillin. In some embodiments, the bacterial infection is caused by a type of Pseudomonas aeruginosa. In some embodiments, the antibacterial agent is an antibacterial agent of the carbapenem class. In certain embodiments, the antibacterial agent is meropenem. 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 subject is a mammal. In certain embodiments, the gelsolin agent comprises plasma gelsolin (pGSN), optionally recombinant pGSN. In some embodiments, the pharmaceutical composition also comprises a pharmaceutically acceptable carrier. In some embodiments, the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of a gelsolin molecule. In certain embodiments, the pharmaceutical composition also includes a pharmaceutically acceptable carrier.

[0010] In yet another aspect of the present invention, a method for treating a viral infection in a subject is provided, comprising administering an effective amount of a gelsolin agent to a subject having a viral infection, wherein the gelsolin agent is administered at least 3, 4, 5, 6, 7, 8, 9, or more days after the subject is infected with the viral infection, and is not administered on the day the subject is infected with the viral infection, one day after the subject is infected with the viral infection, or two days after the subject is infected with the viral infection. In some embodiments, the effective amount of the gelsolin agent increases the therapeutic effect against the viral infection in the subject compared to a control therapeutic effect. In certain embodiments, the control therapeutic effect 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% greater 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 a 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 administering a gelsolin agent reduces the level of viral infection in the subject compared to a control level of viral infection, where the control level of infection comprises the level of infection in the absence of administration of the gelsolin agent. 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 determining means include one or more of assaying, observing the subject, assessing one or more physiological symptoms of viral infection in the subject, and assessing the subject's viability. In some embodiments, the physiological symptoms include one or more of fever, fatigue, weight loss, and death. In some embodiments, the assay includes a means for detecting the presence, absence, and / or level of a characteristic of viral infection in a biological sample from the subject. In certain embodiments, administration of an effective amount of a gelsolin agent increases the subject's viability compared to a control's viability. In some embodiments, the control's viability is the viability in the absence of administration of the gelsolin agent. In certain embodiments, the increase in the subject's viability is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more than the control's viability. %, 95%, 100%, 125%, 150%, 175%, or 200% higher. In some embodiments, the means of administration of the gelsolin agent is selected from oral, sublingual, buccal, intranasal, intravenous, inhalation, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intrasynovial, and intraocular administration. In some embodiments, the subject is a mammal, optionally a human. In certain embodiments, the method also includes treating the subject with an antiviral agent one or more days before administering the gelsolin agent to the subject, wherein the antiviral agent is administered on one or more of the following days: 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, synergistically effective amounts of each of the gelsolin agent and the antiviral agent are administered to the subject and have a synergistic therapeutic effect against the viral infection compared to the therapeutic effect of a control, and the antiviral agent is administered in a clinically tolerated amount. In some embodiments, the control comprises the therapeutic effect of administering a clinically tolerated amount of an antiviral agent without administering a gelsolin agent. In certain embodiments, a clinically tolerated amount of an antiviral agent is an amount less than the maximum tolerated dose (MTD) of the antiviral agent. In some embodiments, the MTD of an antiviral agent is the highest possible, yet tolerable, dose level of the antiviral agent for a subject. In some embodiments, the MTD of the antiviral agent is determined at least in part based on a preselected clinically limiting toxicity for the antiviral agent. In certain embodiments, a synergistically effective amount of a gelsolin agent and an antiviral agent reduces the minimum effective dose (MED) of the antiviral agent in a subject. In some embodiments, the MED is the lowest dose level of the antiviral agent that provides a clinically significant response in mean efficacy, which response is statistically significantly greater than the response provided by a control that does not include that dose of the antibacterial agent. In certain embodiments, the means of administration of the gelsolin agent and the antiviral agent are independently selected from oral, sublingual, buccal, intranasal, inhalation, intravenous, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intrasynovial, and intraocular administration. [Brief explanation of the drawings]

[0011] [Figure 1A] Figure 1A shows a graph of the results of a systemic experiment measuring the improvement of host defense against bacterial pneumonia after pGSN administration. In vitro, pGSN improves macrophage uptake (Figure 1A) when present at 125-250 μg / ml, similar to normal plasma levels. * = p<.05 vs. control, n = 6-12 per group. Serotype 3 Strep. pneumoniae was used in all experiments. [Figure 1B] Figure 1B shows a graph of the results of a systemic experiment measuring the improvement of host defense against bacterial pneumonia after pGSN administration. In vitro, pGSN improves killing of internalized pneumococci (Figure 1B) when present at 125-250 μg / ml, similar to normal plasma levels. * = p < .05 vs. control, n = 6-12 per group. Serotype 3 Strep. pneumoniae was used in all experiments. [Figure 1C] Figure 1C shows a graph of the results of a systemic experiment measuring the improvement of host defense against bacterial pneumonia after pGSN administration. In vivo, pGSN (10 mg sc 2 hours before infection and 8 and 20 hours after infection) improved bacterial clearance (fewer surviving bacteria at 24 hours) in Bl6 mice challenged with 10 pneumococci by insufflation (Figure 1C). * = p < .05 vs. control, n = 6–12 per group. Serotype 3 Strep. pneumoniae was used in all experiments. [Figure 1D] Figure 1D shows a graph of the results of a systemic experiment measuring improved host defense against bacterial pneumonia after pGSN administration. In vivo, similar results to those shown in Figure 1C were observed when pGSN was administered as an aerosol 15 or 30 minutes before infection (Figure 1D). * = p < .05 vs. control, n = 6–12 per group. All experiments used serotype 3 Strep. pneumoniae. [Figure 1E]Figure 1E shows a graph of the results of a systemic experiment measuring improved host defense against bacterial pneumonia after pGSN administration. Systemic pGSN (sc) improved survival from primary pneumococcal pneumonia (Figure 1E, using a 3 x 10 CFU inoculum) even in the absence of antibiotic treatment. * = p < .05 vs. control, n = 6-12 per group. Serotype 3 Strep. pneumoniae was used in all experiments. [Figure 1F] Figure 1F shows a graph of the results of a systemic experiment measuring improved host defense against bacterial pneumonia after pGSN administration. Systemic pGSN (sc) improved survival from secondary post-influenza pneumococcal pneumonia (Figure 1F, using a 500 CFU inoculum on day 7 after mild influenza infection with PR8) even in the absence of antibiotic treatment. * = p < .05 vs. control, n = 6–12 per group. Serotype 3 Strep. pneumoniae was used in all experiments. [Figure 2A] Figure 2A provides a graph demonstrating that NOS3 is required for the effect of pGSN on macrophages. Figure 2A shows the results of experiments demonstrating macrophage killing of pneumococci in vitro. When NOS3-deficient cells or animals were used, macrophage clearance of bacteria was no longer enhanced. *=p<0.01. [Figure 2B] Figure 2B provides a graph showing that NOS3 is required for the effect of pGSN on macrophages. Figure 2B shows the results of an experiment demonstrating macrophage clearance of bacteria in vivo. When NOS3-deficient cells or animals were used, macrophage clearance of bacteria was no longer enhanced. *=p<0.01. [Figure 3A] Figure 3A provides a graph of a study of antibiotic-susceptible pneumococcal pneumonia. Figure 3A shows the results of treatment with pGSN (5 mg i.p. on days 2 and 3 post-infection) demonstrating improved survival in mice infected with serotype 3 pneumococci (*=p=0.01, n=20 / group, summary of two studies, 10 mice per group per study). [Figure 3B]Figure 3B provides a graph of a study of antibiotic susceptibility pneumococcal pneumonia. Figure 3B provides the results of treatment with penicillin (PEN, 100 µg by i.m. on days 2 and 3 post-infection), demonstrating improved survival in mice infected with serotype 3 pneumococci (* = p = .02, n = 8–9 / group, single study). [Figure 4A] Figure 4A provides a graph of the results of a study of antibiotic-resistant pneumococcal pneumonia. The results in Figure 4A demonstrate that treatment with pGSN (5 mg ip daily starting on day 1 postinfection) improved survival in mice infected with serotype 14 pneumococci compared to vehicle or penicillin (PEN, 1 mg dose im daily) (*, p = .02 and .04, respectively, log-rank comparisons after Sidak correction for multiple comparisons). Combination treatment with pGSN and penicillin also resulted in higher survival compared to vehicle or penicillin (**, p = .0001 for both comparisons, log-rank comparisons with Sidak correction for multiple comparisons). After Sidak correction for multiple comparisons, the survival rate of the pGSN vs. pGSN+PEN group was not statistically significant (p = .47 n = 38–41 / group, summary of four trials, 8–11 mice per group per trial). [Figure 4B] Figure 4B provides a graph of the results of a study of antibiotic-resistant pneumococcal pneumonia. (Figure 4B) Assessment of weight loss showed more rapid weight recovery in the pGSN or pGSN+PEN groups (mean values for each day shown, p = .001, ANOVA; n = 38–41 per group across four trials in B; the last observation of any mouse was carried forward after death). [Figure 4C] Figure 4C provides a graph of the results of a study of antibiotic-resistant pneumococcal pneumonia. (Figure 4C) Morbidity assessment showed lower morbidity indices in the pGSN or pGSN+PEN groups (mean values for each day shown, p=0.04, ANOVA; n=30 per group across three trials in C; the last observation of any mouse was carried forward after death). [Figure 5]A table of data results from nine experiments in which four delayed-administration treatments were evaluated is provided. Figure 5 shows details of the nine experiments, including the pilot and distance measurement studies. Column H shows the change in bacterial growth method for the superior growth results obtained using the 2-fold growth method in BHI broth for penicillin-resistant Streptococcus pneumoniae [Restrepo AV et al., BMC Microbiol 2005;5:34]. The data provided in the table show that in all nine experiments, survival was highest in the PEN + pGSN group. Survival in the PEN + pGSN group was higher than in the pGSN group, and both survival rates were higher than in the vehicle or PEN alone groups. The difference in survival was statistically significant as determined by a pooled analysis of all nine studies using log-rank analysis with Sidak correction for multiple comparisons. Detailed results from the statistical analysis of the final four experiments (numbers 6–9) are summarized in Figure 4A–C. [Figure 6] Data from three experiments assessing survival after administration of meropenem doses with and without rhu-pGSN to neutropenic mice are provided in the table. Figure 6 details three experiments in which the indicated meropenem doses were administered subcutaneously beginning 3 hours after infection with MDR Pseudomonas aeruginosa and then every 8 hours for 5 days. Meropenem doses were administered either with or without rhu-pGSN. rhu-pGSN was administered as a 12 mg intraperitoneal injection on days -1, 0, 1, 2, 3, 4, and 5. n / N = number of surviving mice / number of mice treated. [Figure 7A]Figure 7A provides a graph demonstrating the survival benefit observed with combination treatment of meropenem and rhu-pGSN. BALB / c-Cy mice rendered neutropenic with cyclophosphamide were infected with the UNC-D strain of Pseudomonas aeruginosa and treated with either meropenem alone (1250 mg / kg / day subcutaneously three times daily from 3 hours postinfection for 5 days) or in combination with pGSN (12 mg / day intraperitoneally daily for days -1 to +5). Mice were euthanized when the endpoint criteria were reached or at the end of the study on day 7. Survival analysis was performed by log-rank test using two studies with a group size of n = 8 (Figures 7A and 7B). Control mortality on day 7 was ≥ 50% with the same 1250 mg meropenem dose. p values indicate the survival advantage of combination therapy over meropenem alone. MTD refers to the mean time to death. [Figure 7B] Figure 7B provides a graph demonstrating the survival benefit observed with combination treatment of meropenem and rhu-pGSN. BALB / c-Cy mice rendered neutropenic with cyclophosphamide were infected with the UNC-D strain of Pseudomonas aeruginosa and treated with either meropenem alone (1250 mg / kg / day subcutaneously three times daily from 3 hours postinfection for 5 days) or in combination with pGSN (12 mg / day intraperitoneally daily from days -1 to +5). Mice were euthanized when the endpoint criteria were reached or at the end of the study period (day 7). Survival analysis was performed by log-rank test using two studies with a group size of n = 8 (Figures 7A and 7B). Control mortality at day 7 was ≥ 50% with the same 1250 mg meropenem dose. p values indicate the survival advantage of combination therapy over meropenem alone. MTD indicates the mean time to death. [Figure 7C]Figure 7C provides a graph demonstrating the survival benefit observed with combination treatment of meropenem and rhu-pGSN. BALB / c-Cy mice rendered neutropenic with cyclophosphamide were infected with the UNC-D strain of Pseudomonas aeruginosa and treated with either meropenem alone (1250 mg / kg / day subcutaneously three times daily from 3 hours postinfection for 5 days) or in combination with pGSN (12 mg / day intraperitoneally daily from days -1 to +5). Mice were euthanized when the endpoint criteria were reached or at day 7 of the study. The results from the two separate studies in Figure 7A and Figure 7B were combined and analyzed (Figure 7C). p values indicate the survival advantage of combination therapy over meropenem alone. MTD indicates the mean time to death. [Figure 8]Figure 8A provides a graph showing that administration of rhu-pGSN reduces bacterial counts in the lungs. Two studies were performed in which BALB / c-Cy mice were infected with the UNC-D strain of Pseudomonas aeruginosa and treated with either meropenem alone (1250 mg / kg / day subcutaneously three times daily from 3 hours postinfection for 5 days) or in combination with pGSN (12 mg / day intraperitoneally daily for days -1 to +5). Mice were euthanized when they reached the endpoint criteria (open circles) or survivors at day 7 of the study (filled circles). Bacteria were counted from homogenized lungs by plate count. Figure 8A shows a graph of the results of the first study; p values refer to unpaired Student's t-test comparisons of the combination therapy versus meropenem alone. The line at the bottom of the graph indicates the limit of detection. Figure 8B provides a graph showing that administration of rhu-pGSN reduces bacterial counts in the lungs. Two studies were performed in which BALB / c-Cy mice were infected with the UNC-D strain of Pseudomonas aeruginosa and treated with either meropenem alone (1250 mg / kg / day subcutaneously three times daily from 3 hours postinfection for 5 days) or in combination with pGSN (12 mg / day intraperitoneally daily for days -1 to +5). Mice were euthanized when the endpoint criteria were reached (open circles) or survivors at day 7 of the study (filled circles). Bacteria were enumerated from homogenized lungs by plate counting. Figure 8B shows a graph of the results from the second study. Individual and combined data were analyzed across the two studies, and pairwise analyses were performed for meropenem therapy alone (Mero) versus in combination with pGSN. p values refer to unpaired Student's t-test comparisons of combination therapy versus meropenem alone. The line at the bottom of the graph indicates the limit of detection. Figure 8C shows a graph of the combined data from the two studies shown in Figures 8A and 8B. p values refer to unpaired Student's t-test comparisons of the combination therapy versus meropenem alone. The line at the bottom of the graph indicates the limit of detection. [Figure 9A]Figures 9A–C provide graphs demonstrating that rhu-pGSN limits infection-induced lung injury. Two studies were performed in which BALB / c-Cy mice were infected with the UNC-D strain of Pseudomonas aeruginosa and treated with either meropenem alone (1250 mg / kg / day subcutaneously three times daily from 3 hours postinfection for 5 days) or in combination with pGSN (12 mg / day intraperitoneally daily for days -1 to +5). Mice were euthanized when they reached the endpoint criteria (open circles) or survivors at day 7 of the study (filled circles). Representative sections of the lungs were excised and processed for H&E staining and scoring. Figure 9A shows a graph of the results from the first study; data were analyzed separately for the two individual studies and combined in a pairwise analysis of meropenem therapy alone (Mero) or in combination with pGSN. p values refer to unpaired Student's t-test comparison of combination therapy versus meropenem alone. [Figure 9B] Figures 9A–C provide graphs demonstrating that rhu-pGSN limits infection-induced lung injury. Two studies were performed in which BALB / c-Cy mice were infected with the UNC-D strain of Pseudomonas aeruginosa and treated with either meropenem alone (1250 mg / kg / day subcutaneously three times daily from 3 hours postinfection for 5 days) or in combination with pGSN (12 mg / day intraperitoneally daily for days -1 to +5). Mice were euthanized when they reached the endpoint criteria (open circles) or survivors at day 7 of the study (filled circles). Representative sections of the lungs were excised and processed for H&E staining and scoring. Figure 9B shows a graph of the results from the second study. Data were analyzed separately for the two individual studies and combined in a pairwise analysis of meropenem therapy alone (Mero) or in combination with pGSN. p values refer to unpaired Student's t-test comparison of combination therapy versus meropenem alone. [Figure 9C] Figures 9A-C provide graphs showing that rhu-pGSN limits infection-induced lung injury. Figure 9C shows a graph of the combined data from the two studies shown in Figures 9A and 9B. p values refer to unpaired Student's t-test comparisons of the combination therapy versus meropenem alone. [Figure 10] Overall survival data with mild lung injury from surviving mice treated with different doses of meropenem are shown in the table. The indicated meropenem doses were administered subcutaneously starting 3 hours post-infection and then every 8 hours for 5 days. rhu-pGSN was administered as a 12 mg intraperitoneal injection on days -1, 0, 1, 2, 3, 4, and 5. An asterisk (*) indicates that a total of three mice (all from experiment 2) were euthanized 20 hours post-challenge but did not have lung injury; one mouse was present in each of the three meropenem + rhu-pGSN treatment groups. Removing these three mice from the rhu-pGSN count resulted in a final count of 41 / 61 (67.2%). n / N = number of surviving mice with a combined lung injury score of ≤2 / number of treated mice. [Figure 11A] Figure 11A shows graphs of two experiments demonstrating the recovery of baseline temperatures in mice treated with either meropenem alone or meropenem plus rhu-pGSN. In both experiments, BALB / c-Cy mice were infected with the UNC-D strain of Pseudomonas aeruginosa. Figure 11A shows a graph of the temperature of mice in the first experiment treated with meropenem alone (1250 mg / kg / day subcutaneously every 8 hours from 3 hours post-infection for 5 days). Animal temperatures were monitored every 8 hours post-infection until the end of the study. Mice were euthanized when the endpoint criteria were reached (open circles) or at the end of the study on day 7 (filled circles). [Figure 11B] Figure 11B shows graphs of two experiments demonstrating the recovery of baseline temperatures in mice treated with either meropenem alone or meropenem plus rhu-pGSN. In both experiments, BALB / c-Cy mice were infected with the UNC-D strain of Pseudomonas aeruginosa. Figure 11B shows a graph of the temperature of mice in the first experiment treated with meropenem in combination with rhu-pGSN (12 mg / day intraperitoneally daily from day -1 to day +5). Animal temperatures were monitored every 8 hours post-infection until the end of the study. Mice were euthanized when the endpoint criteria were reached (open circles) or at the end of the study on day 7 (filled circles). [Figure 11C] Figure 11C shows graphs of two experiments demonstrating the recovery of baseline temperatures in mice treated with either meropenem alone or meropenem plus rhu-pGSN. In both experiments, BALB / c-Cy mice were infected with the UNC-D strain of Pseudomonas aeruginosa. Figure 11C shows a graph of the temperature of mice in the second experiment treated with meropenem alone (same regimen as in Figure 11A). Animal temperatures were monitored every 8 hours post-infection until the end of the study. Mice were euthanized when the endpoint criteria were reached (open circles) or at the end of the study on day 7 (filled circles). [Figure 11D] Figure 11D shows graphs of two experiments demonstrating the recovery of baseline temperatures in mice treated with either meropenem alone or meropenem plus rhu-pGSN. In both experiments, BALB / c-Cy mice were infected with the UNC-D strain of Pseudomonas aeruginosa. Figure 11D shows a graph of the temperature of mice in a second experiment treated with meropenem in combination with rhu-pGSN (the same regimen as in Figure 11B). Animal temperatures were monitored every 8 hours post-infection until the end of the study. Mice were euthanized when the endpoint criteria were reached (open circles) or at the end of the study on day 7 (filled circles). [Figure 12-1] A table showing details of a therapeutic trial using recombinant human plasma gelsolin (rhu-pGSN) in murine influenza is provided. * = Treatment effect was scored as "positive" if % survival with pGSN vs. vehicle was ≥ 10% better; negative if survival with pGSN was < 10%. [Figure 12-2] A table showing details of a therapeutic trial using recombinant human plasma gelsolin (rhu-pGSN) in murine influenza is provided. * = Treatment effect was scored as "positive" if % survival with pGSN vs. vehicle was ≥ 10% better; negative if survival with pGSN was < 10%. [Figure 12-3]A table showing details of a therapeutic trial using recombinant human plasma gelsolin (rhu-pGSN) in murine influenza is provided. * = Treatment effect was scored as "positive" if % survival with pGSN vs. vehicle was ≥ 10% better; negative if survival with pGSN was < 10%. [Figure 13] Figure 13 provides a summary of survival data using various treatment regimens. pGSN is plasma gelsolin. [Figure 14A] Figures 14A-H provide survival and morbidity analyses of various treatment regimens. Comparison of survival (Figures 14A, C, E, and G) and morbidity (Figures 14B, D, F, and H) rates for mice treated with rhu-pGSN or vehicle. (Figures 14A-B) Results from all 18 studies using delayed treatment (typically 10 or more mice per group; see details in Figures 12 and 13). In some studies, treatment was initiated on day 6 or day 3 with different treatment groups. (Figures 14C-D) Results from 13 studies using delayed treatment starting on day 6 or later. (Figures 14E-F) Results from 8 studies using treatment starting on day 3. (Figures 14G-H) Results from 4 studies starting at a low dose on day 3 and increasing the dose on days 6 / 7. *=0.000001, 0.00001, 0.0005, 0.0005 for Figures 14A, C, E, and G, respectively; p<0.0001 for Figures 14B, D, F, and H. [Figure 14B]Figures 14A-H provide survival and morbidity analyses of various treatment regimens. Comparison of survival (Figures 14A, C, E, and G) and morbidity (Figures 14B, D, F, and H) rates for mice treated with rhu-pGSN or vehicle. (Figures 14A-B) Results from all 18 studies using delayed treatment (typically 10 or more mice per group; see details in Figures 12 and 13). In some studies, treatment was initiated on day 6 or day 3 with different treatment groups. (Figures 14C-D) Results from 13 studies using delayed treatment starting on day 6 or later. (Figures 14E-F) Results from 8 studies using treatment starting on day 3. (Figures 14G-H) Results from 4 studies starting at a low dose on day 3 and increasing the dose on days 6 / 7. *=0.000001, 0.00001, 0.0005, 0.0005 for Figures 14A, C, E, and G, respectively; p<0.0001 for Figures 14B, D, F, and H. [Figure 14C] Figures 14A-H provide survival and morbidity analyses of various treatment regimens. Comparison of survival (Figures 14A, C, E, and G) and morbidity (Figures 14B, D, F, and H) rates for mice treated with rhu-pGSN or vehicle. (Figures 14A-B) Results from all 18 studies using delayed treatment (typically 10 or more mice per group; see details in Figures 12 and 13). In some studies, treatment was initiated on day 6 or day 3 with different treatment groups. (Figures 14C-D) Results from 13 studies using delayed treatment starting on day 6 or later. (Figures 14E-F) Results from 8 studies using treatment starting on day 3. (Figures 14G-H) Results from 4 studies starting at a low dose on day 3 and increasing the dose on days 6 / 7. *=0.000001, 0.00001, 0.0005, 0.0005 for Figures 14A, C, E, and G, respectively; p<0.0001 for Figures 14B, D, F, and H. [Figure 14D]Figures 14A-H provide survival and morbidity analyses of various treatment regimens. Comparison of survival (Figures 14A, C, E, and G) and morbidity (Figures 14B, D, F, and H) rates for mice treated with rhu-pGSN or vehicle. (Figures 14A-B) Results from all 18 studies using delayed treatment (typically 10 or more mice per group; see details in Figures 12 and 13). In some studies, treatment was initiated on day 6 or day 3 with different treatment groups. (Figures 14C-D) Results from 13 studies using delayed treatment starting on day 6 or later. (Figures 14E-F) Results from 8 studies using treatment starting on day 3. (Figures 14G-H) Results from 4 studies starting at a low dose on day 3 and increasing the dose on days 6 / 7. *=0.000001, 0.00001, 0.0005, 0.0005 for Figures 14A, C, E, and G, respectively; p<0.0001 for Figures 14B, D, F, and H. [Figure 14E] Figures 14A-H provide survival and morbidity analyses of various treatment regimens. Comparison of survival (Figures 14A, C, E, and G) and morbidity (Figures 14B, D, F, and H) rates for mice treated with rhu-pGSN or vehicle. (Figures 14A-B) Results from all 18 studies using delayed treatment (typically 10 or more mice per group; see details in Figures 12 and 13). In some studies, treatment was initiated on day 6 or day 3 with different treatment groups. (Figures 14C-D) Results from 13 studies using delayed treatment starting on day 6 or later. (Figures 14E-F) Results from 8 studies using treatment starting on day 3. (Figures 14G-H) Results from 4 studies starting at a low dose on day 3 and increasing the dose on days 6 / 7. *=0.000001, 0.00001, 0.0005, 0.0005 for Figures 14A, C, E, and G, respectively; p<0.0001 for Figures 14B, D, F, and H. [Figure 14F]Figures 14A-H provide survival and morbidity analyses of various treatment regimens. Comparison of survival (Figures 14A, C, E, and G) and morbidity (Figures 14B, D, F, and H) rates for mice treated with rhu-pGSN or vehicle. (Figures 14A-B) Results from all 18 studies using delayed treatment (typically 10 or more mice per group; see details in Figures 12 and 13). In some studies, treatment was initiated on day 6 or day 3 with different treatment groups. (Figures 14C-D) Results from 13 studies using delayed treatment starting on day 6 or later. (Figures 14E-F) Results from 8 studies using treatment starting on day 3. (Figures 14G-H) Results from 4 studies starting at a low dose on day 3 and increasing the dose on days 6 / 7. *=0.000001, 0.00001, 0.0005, 0.0005 for Figures 14A, C, E, and G, respectively; p<0.0001 for Figures 14B, D, F, and H. [Figure 14G] Figures 14A-H provide survival and morbidity analyses of various treatment regimens. Comparison of survival (Figures 14A, C, E, and G) and morbidity (Figures 14B, D, F, and H) rates for mice treated with rhu-pGSN or vehicle. (Figures 14A-B) Results from all 18 studies using delayed treatment (typically 10 or more mice per group; see details in Figures 12 and 13). In some studies, treatment was initiated on day 6 or day 3 with different treatment groups. (Figures 14C-D) Results from 13 studies using delayed treatment starting on day 6 or later. (Figures 14E-F) Results from 8 studies using treatment starting on day 3. (Figures 14G-H) Results from 4 studies starting at a low dose on day 3 and increasing the dose on days 6 / 7. *=0.000001, 0.00001, 0.0005, 0.0005 for Figures 14A, C, E, and G, respectively; p<0.0001 for Figures 14B, D, F, and H. [Figure 14H]Figures 14A-H provide survival and morbidity analyses of various treatment regimens. Comparison of survival (Figures 14A, C, E, and G) and morbidity (Figures 14B, D, F, and H) rates for mice treated with rhu-pGSN or vehicle. (Figures 14A-B) Results from all 18 studies using delayed treatment (typically 10 or more mice per group; see details in Figures 12 and 13). In some studies, treatment was initiated on day 6 or day 3 with different treatment groups. (Figures 14C-D) Results from 13 studies using delayed treatment starting on day 6 or later. (Figures 14E-F) Results from 8 studies using treatment starting on day 3. (Figures 14G-H) Results from 4 studies starting at a low dose on day 3 and increasing the dose on days 6 / 7. *=0.000001, 0.00001, 0.0005, 0.0005 for Figures 14A, C, E, and G, respectively; p<0.0001 for Figures 14B, D, F, and H. [Figure 15] Experimental results are provided showing the top 50 differentially expressed genes that were up- and downregulated in lung tissue from animals treated with vehicle or rhu-pGSN (day 9). Heatmap (right scale showing range -2 to +2) showing the top 50 down-regulated (left) and up-regulated (right) genes in the lungs of rhu-pGSN-treated animals on day 9. [Figure 16-1] Figure 1 shows the top 10 downregulated gene ontology (GO) processes and pathways in lung tissue (day 9) treated with plasma gelsolin (pGSN). [Figure 16-2] Figure 1 shows the top 10 downregulated gene ontology (GO) processes and pathways in lung tissue (day 9) treated with plasma gelsolin (pGSN). DETAILED DESCRIPTION OF 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 with a microbial infection can result in a synergistic therapeutic effect of the two agents that reduces the microbial infection. In some embodiments, the present invention relates to exogenous gelsolin agents and antibacterial agents that, when administered to a subject with a microbial infection, act synergistically in the subject, such that their synergistic effect results in a therapeutic effect that is greater than the therapeutic effect of administering to the subject either the gelsolin agent or the antibacterial agent at a clinically tolerated dose, without administering the other agent to the subject. Certain methods of the invention include administering a pharmaceutical composition of the 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 invention include administering a delayed dose of a gelsolin agent to a subject having a viral infection, which enhances treatment of the viral infection in the subject.

[0013] Synergistic therapeutic effects The methods of the present invention include producing a synergistic therapeutic effect in a subject with a microbial infection to alleviate and treat the microbial infection. It has been concluded that even if one or both of a gelsolin agent and an antibacterial agent do not have a statistically significant individual therapeutic effect on the microbial infection, they can be administered in combination with each other to produce a synergistic therapeutic effect on the microbial infection. Thus, in some embodiments of the present invention, a microbial infection in a subject caused by a microorganism 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] The term "individual therapeutic effect" as used herein with respect to an agent, such as a gelsolin agent or an antibacterial agent, refers to the therapeutic effect of the agent when administered to a subject with a microbial infection. With respect to the methods and compositions of the present invention, the individual therapeutic effect of a gelsolin agent is the therapeutic effect against a microbial infection in a subject that results from administering the gelsolin agent to a 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 against a microbial infection in a subject that results from administering the antibacterial agent to a subject without administering the gelsolin agent.

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

[0016] In some embodiments, the methods of the present invention comprise administering synergistically effective amounts of a gelsolin agent and a non-therapeutic antibacterial agent to a subject with a microbial infection. The synergistic effect of combined administration may enhance the therapeutic effect of the antibacterial agents. The term "non-therapeutic agent" is used herein in reference 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 a non-therapeutic agent used in connection with the methods and compositions of the present invention is not an antibacterial agent referred to in the art as a "therapeutic agent" or "antibacterial therapeutic agent." For example, an antibacterial agent that does not have a statistically significant individual therapeutic effect against a microbial infection when administered in a clinically acceptable amount to a medical professional would not be designated as a therapeutic agent for administration to a subject with that microbial infection. Similarly, it is recognized in the art that penicillin does not have a statistically significant individual therapeutic effect against certain microbial infections, and therefore, penicillin would be understood and defined as a "non-therapeutic agent" with respect to those infections. In certain embodiments of the present invention, an 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, an antimicrobial agent is a non-therapeutic agent with respect to its individual therapeutic effect on an antimicrobial-resistant microbial infection in a subject. Gelsolin agents may be referred to herein as non-therapeutic agents with respect to microbial infections.

[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, a gelsolin agent may have an individual therapeutic effect against a microbial infection, while an 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 a microbial infection may or may not be due to antibacterial resistance of the microorganism causing the microbial infection. As used herein, the term "resistant" in relation to a microorganism or microbial infection refers to a microorganism that is not killed or reduced, respectively, by an antibacterial agent. In some embodiments of the present invention, the individual therapeutic effect of the antibacterial agent against an antibacterial-resistant microorganism or infection may be zero.

[0018] In certain circumstances, a microbial infection in a subject is caused by a microorganism that is resistant to the therapeutic effect of a particular antimicrobial agent. Acquired antimicrobial resistance can be understood as the ability of a disease-causing microorganism to withstand exposure to an antimicrobial agent that was previously an effective treatment for the disease. An "antimicrobial-resistant" microorganism may be the cause of a microbial infection in a subject, and one or more antimicrobial agents, including one or more antimicrobial agents 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 caused by the presence of Streptococcus pneumoniae in the subject that is resistant to the therapeutic effect of one or more antibiotics.

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

[0020] In some embodiments of the present invention, a clinically acceptable amount of an antibacterial agent is an amount less than the maximum tolerated dose (MTD) of the antibacterial agent. It is understood in the art how to determine an individual's MTD to prevent or reduce negative side effects of administering an agent. In some embodiments of the present invention, the MTD of an antibacterial agent is the highest possible dose level of the antibacterial agent for a 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, subject discomfort, physiological distress, an increased risk of subject death, etc. In some embodiments of the present invention, the MTD of an antibacterial agent administered to a subject is determined at least in part based on a preselected clinically limiting toxicity for the antibacterial agent. For example, a dose or amount of an antibacterial agent that is effective in reducing or killing microorganisms resistant to the antibacterial agent may, when administered to a subject, result in clinically unacceptable toxicity and / or harmful side effects for the subject.

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

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

[0023] [Table 1]

[0024] Therapeutic Compositions and Methods The synergistic therapeutic effect of a composition of the invention or a treatment method of the invention (also referred to herein as a "response" to a treatment method of the invention) can be determined by detecting one or more physiological effects of the treatment, such as, for example, a reduction or absence of symptoms following administration of a synergistic treatment. Additional means for monitoring and assessing microbial infection in a subject and determining one or more of the presence, absence, level, severity, change in severity, etc. of a microbial infection in a subject in response 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 a synergistic combination of a gelsolin agent and an antibacterial agent to a subject with a microbial infection, each in an amount effective to produce a synergistic therapeutic effect for reducing the microbial infection in the subject. The gelsolin agent and the antibacterial agent can be administered simultaneously. The gelsolin agent and the antibacterial agent can be administered in the same formulation or in separate formulations that are administered simultaneously in the subject.

[0026] The methods and compositions of the present invention can be used to treat microbial infections. As used herein, the terms "treat," "treated," or "treating" when used in reference to a microbial infection can refer to prophylactic treatment that reduces the likelihood that a subject will develop a microbial infection, and can also refer to treatments that eliminate or ameliorate a microbial infection, preventing a microbial infection from becoming more advanced. The term "microbial infection" can be used to refer to treatment after a subject has developed a microbial infection to prevent the infection from progressing or becoming severe and / or to slow the progression of the microbial infection compared to the progression of the microbial infection in the absence of the therapeutic methods of the present invention. Gelsolin Gelsolin is a highly conserved, multifunctional protein initially described in the cytosol of macrophages and subsequently identified in many vertebrate cell types (Piktel E. et al., Int J Mol Sci 2018;19:E2516; Silacci P. et al., Cell Mol Life Sci 2004;61:2614-23). A unique property of gelsolin is that its gene expresses a splice variant encoding a distinct plasma isoform (pGSN) that is secreted into the extracellular fluid and differs from its cytoplasmic counterpart (cGSN) by expressing an additional 25 amino acid sequence. pGSN typically circulates in mammalian blood at concentrations of 200-300 μg / ml, making it one of the most abundant plasma proteins. As used herein, the term "gelsolin agent" refers to a composition containing a gelsolin molecule, its functional fragment, or a functional derivative of the gelsolin molecule. In some embodiments of the present invention, the gelsolin agent comprises only one or more of the gelsolin molecule, its functional fragment, or functional derivative of the gelsolin molecule.In certain embodiments of the present invention, the gelsolin agent can comprise one of additional components, such as, but not limited to, 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 molecule included in the compositions and methods of the present invention can be a recombinant gelsolin molecule.

[0027] As used herein, the term "gelsolin agent" refers to a compound containing an exogenous gelsolin molecule. As used herein, the term "exogenous" in reference to a gelsolin molecule refers to a gelsolin molecule that is administered to a subject, even if the same gelsolin molecule naturally exists in the subject, which may be referred to as an endogenous gelsolin molecule. The gelsolin agent included in the method or composition of the present invention may be a wild-type gelsolin molecule (GenBank accession number: X04412), an isoform, an analog, a variant, a fragment or a functional derivative of a gelsolin molecule.

[0028] Some embodiments of the present invention may include "gelsolin analogs," which, as used herein, refer to compounds whose function is substantially similar to either native gelsolin or a fragment thereof. A gelsolin analog comprises a biologically active amino acid sequence substantially similar to the gelsolin sequence and may have substitutions, deletions, extensions, replacements, or otherwise modified sequences that have a biological activity substantially similar to that of gelsolin. For example, a gelsolin analog 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 skilled in the art.

[0029] Certain embodiments of the methods and compositions of the present invention include fragments of gelsolin molecules. The term "fragment" is intended to include any portion of a gelsolin molecule that provides a segment of gelsolin that maintains at least some or substantially all of the level of biological activity of the "parent" gelsolin; this term is intended to include 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 refers to the gelsolin molecule from which the fragment or derivative sequence 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 some to all of the functions of its parent gelsolin molecule. In some embodiments, the methods and compositions of the present invention may include a "variant" of gelsolin. As used herein, a gelsolin variant may 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, a gelsolin variant is referred to as a functional variant and retains at least some to all of the functions of its parent gelsolin molecule.

[0031] Gelsolin derivatives are also contemplated for inclusion in the methods and compositions of the present invention. A "functional derivative" of gelsolin is a derivative that has biological activity substantially similar to that of gelsolin. "Substantially similar" means that the activity may be qualitatively the same, but quantitatively different. For example, a functional derivative of gelsolin may contain the same amino acid backbone as gelsolin, but may also contain other modifications, such as post-translational modifications such as attached phospholipids or covalently attached carbohydrates, depending on the necessity of such modifications for the performance of the therapeutic methods of the present invention. As used herein, this term also includes chemical derivatives of gelsolin. Such derivatives may improve the solubility, absorption, biological half-life, etc. of gelsolin. Derivatives may also reduce the toxicity of gelsolin or eliminate or mitigate 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, edited by 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 disease-causing microorganisms, sometimes referred to as "microbial infections." 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 cause bacterial, fungal, viral, and parasitic infections, respectively. The term "antimicrobial agent," as used herein with respect to treating or reducing an infection in a subject, includes antibacterial, antifungal, antiviral, and antiparasitic agents that can be administered to a subject to treat bacterial, fungal, viral, and parasitic infections, respectively. The present invention, in some aspects, includes methods for treating an infection in a subject. In some embodiments of the invention, the subject is known to have a microbial infection, is suspected of being exposed to, is 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 fatigue, body temperature, fever, cough, pulmonary exudate, 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 differ from the characteristics of the same microbial infection in another animal species. The characteristics present in different microbial infections and the characteristics present in humans and / or animals are known in the art. Those skilled in the art can easily 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 "characteristics" 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 a disorder 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 that occur in a subject due to infectious organisms such as microorganisms, including, but not limited to, bacteria, viruses, parasites, fungi, and protozoans.

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

[0036] Bacteria that can cause a 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, and Streptococcus species, such as Group A Streptococcus pyogenes. pyogenes, Streptococcus viridans group, Group B hemolytic streptococci (Streptococcus agalactiae), Streptococcus bovis, anaerobic streptococcal species, Streptococcus pneumoniae, and Streptococcus faecalis, Bacillus species, e.g., Bacillus anthracis, Corynebacterium species, e.g., Corynebacterium diphtheriae, aerobic Corynebacterium species, and anaerobic Corynebacterium species, Diphtheroids species, Listeria species, e.g., Listeria monocytogenes monocytogenes, Erysipelothrix species such as Erysipelothrix rhusiopathiae, and 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, and Klebsiella species, such as Klebsiella pneumoniae. pneumoniae, Salmonella spp., Shigella spp., Serratia spp., Acinetobacter spp.; Haemophilus spp., e.g., Haemophilus influenzae, and and Haemophilus ducreyi, Brucella spp., Yersinia spp., e.g., Yersinia pestis and Yersinia enterocolitica, Francisella spp., e.g., Francisella tularensis, Pasteurella spp., e.g., Pasteurella multocida, Vibrio cholerae, Flavobacterium spp., Meningosepticum, Campylobacter spp., e.g., Campylobacter jejuni jejuni, Bacteroides spp. (oral cavity, pharynx) e.g., Bacteroides fragilis, Fusobacterium spp. e.g., Fusobacterium nucleatum, Calymmatobacterium granulomatis, Streptobacillus spp. e.g., Streptobacillus moniliformis, Legionella spp. e.g., Legionella pneumophila.

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

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

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

[0041] Viral agents that, when present in a subject, can cause a viral infection 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, calciviruses, togaviruses, equine encephalitis virus, rubella virus, flaviviruses, dengue virus, Encephalitis viruses, yellow fever viruses, coronaviruses, rhabdoviruses, vesicular stomatitis viruses, rabies viruses, filoviruses, Ebola viruses, paramyxoviruses, parainfluenza viruses, mumps viruses, measles viruses, respiratory syncytial viruses (RSV), orthomyxoviruses, influenza viruses, bunyaviruses, Hantaan viruses, phleboviruses and nairoviruses, arenaviruses, hemorrhagic fever viruses, reoviruses, orbiviruses, rotaviruses, birnaviruses, hepadnaviruses, hepatitis B viruses, and paramyxoviruses. These include, but are not limited to, ruboviruses, papovaviruses, papillomaviruses, polyomaviruses, adenoviruses, herpesviruses including herpes simplex virus type 1 and herpes simplex virus type 2, varicella-zoster virus, poxviruses, variola 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.

[0042] Fungal agents that, when present in a subject, can cause a fungal infection include Cryptococcus species, e.g., Cryptococcus neoformans, Histoplasma species, e.g., Histoplasma capsulatum, Coccidioides species, e.g., Coccidiodes immitis, Paracoccidioides species, e.g., Paracoccidioides brasiliensis, Blastomyces species, e.g., Blastomyces dermatitidis, and Chlamydia species, e.g., Chlamydia trachomatis. trachomatis, Candida species such as Candida albicans, Sporothrix species such as Sporothrix schenckii, Aspergillus species, and fungi of mucormycosis.

[0043] Parasitic agents that, when present in a subject, can cause a parasitic infection can include Plasmodium species, including, for example, Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, and Plasmodium vivax, and Toxoplasma gondii. Blood-borne and / or tissue parasites include Plasmodium spp., Babesia spp., such as Babesia microti and Babesia divergens, Leishmania spp., such as Leishmania tropica, Leishmania spp., (Leishmania braziliensis, Leishmania donovani), Trypanosoma spp., 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 widely described in the literature; see, for example, C.G.A. Thomas, Medical Microbiology, Bailliere Tindall, Great Britain, 1983, the entire contents of which are hereby incorporated by reference. Certain embodiments of the methods and compositions of the present invention can be used to treat infections by these and other medically relevant microorganisms.

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

[0046] A large class of antibacterial agents is antibiotics. Antibiotics that are effective at killing or inhibiting a wide range of bacteria are called broad-spectrum antibiotics. Other types of antibiotics are primarily effective against bacteria from the gram-positive or gram-negative classes. These types of antibiotics are called narrow-spectrum antibiotics. Other antibiotics that are effective against a single organism or disease and not other types of bacteria are called limited-spectrum antibiotics. Antibacterial agents are sometimes classified based on their primary mechanism of action. In general, antibacterial agents are cell wall synthesis inhibitors, cell membrane inhibitors, protein synthesis inhibitors, nucleic acid synthesis or function inhibitors, and competitive inhibitors.

[0047] Antibacterial agents include, but are not limited to, aminoglycosides, beta-lactams, cephalosporins, macrolides, penicillins, quinolones, sulfonamides, and tetracyclines. Examples of antibacterial agents include acedapsone, acetosulfone sodium, alamethicin, alexidine, amdinocillin clavulanate potassium, amdinocillin, amdinocillin pivoxil, amicycline, amifloxacin, amifloxacin mesylate, amikacin, amikacin sulfate, aminosalicylic acid, aminosalicylic acid sodium, amoxicillin, amphomycin, ampicillin, ampicillin sodium, apalcillin sodium, apramycin, aspartocin, Astromycin Sulfate, Avilamycin, Avoparcin, Azithromycin, Azlocillin, Azlocillin Sodium, Bacampicillin Hydrochloride, Bacitracin, Bacitracin Methylenedisalicylate, Zinc Bacitracin, Bambermycin, Benzoylpas Calcium, Berythromycin, Betamycin Sulfate, Biapenem, Biniramycin, Biphenamine Hydrochloride, Bispyrithione MagsulfexMagsulfex, buticacin, butirosin sulfate, capreomycin sulfate, carbadox, carbenicillin disodium, carbenicillin indanyl sodium, carbenicillin phenylsodium, carbenicillin potassium, carumonam sodium, cefaclor, cefadroxil, cefamandole, cefamandole nafate, cefamandole sodium, cefaparol, cefatrizine, cefazaflur sodium, cefazolin, cefazolin sodium, cefbuperazone, cefdinir, cefditoren pivoxil, 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, ceftibuten, ceftizoxime sodium, ceftriaxone sodium, cefuroxime, cefuroxime axetil, cefuroxime pivoxetil, cefuroxime sodium, cephacetrile sodium, cephalexin, cephalexin hydrochloride, cephaloglycin, cephaloridine, cephalothin sodium, cephapirin sodium, cephradine, cetocycline hydrochloride ( Cetocycline Hydrochloride, Cetofenicol, Chloramphenicol, Chloramphenicol Palmitate, Chloramphenicol Pantothenate Complex, Chloramphenicol Sodium Succinate, Chlorhexidine Phosphanilate, Chloroxylenol, Chlortetracycline Bisulfate, Chlortetracycline Hydrochloride, Cilastatin, Cinoxacin, Ciprofloxacin, Ciprofloxacin Hydrochloride, Ciloremycin, Clarithromycin, Potassium Clavulanate, Clinafloxacin Hydrochloride, Clindamycin, Clindamycin Dextrose, Clindamycin Hydrochloride, Clindamycin Palmitate Hydrochloride, Clindamycin Phosphate, Clofazimine, Cloxacillin B Inzatin, Cloxacillin Sodium, Cloxyquin, Colistimethate, Colitimethate Sodium, Colistin Sulfate, Coumermycin, Coumermycin Sodium, Cyclacillin, Cycloserine, Dalfopristin, Dapsone, Daptomycin, Demeclocycline, Demeclocycline Hydrochloride, Demecycline, Denofungin, Diaveridine, Dicloxacillin, Dicloxacillin Sodium um, dihydrostreptomycin sulfate, dipyrithione, dirithromycin, doxycycline, doxycycline calcium, doxycycline phosphatex, doxycycline hyclate, doxycycline monohydrate, droxacin sodium, enoxacin, epicillin, epitetracycline hydrochloride, ertapenem, erythromycin, erythromycin acyslate, erythromycin estolate, erythromycin ethylsuccinate, erythromycin single receptor Erythromycin lactobionate, erythromycin propionate, erythromycin stearate, ethambutol hydrochloride, ethionamide, fleroxacin, floxacillin, fludalanine, flumequine, fosfomycin, fosfomycin tromethamine, fumoxicillin, furazolium chloride, furazolium tartrate, sodium fusidate, fusidic acid, gatifloxacin, gemifloxacin, gentamicin sulfate, gloximonam, gramicidin,Haloprogin, Hetacillin, Hetacillin potassium, Hexedin, Ibafloxacin, Imipenem, Isoconazole, Isepamicin, Isoniazid, Josamycin, Kanamycin sulfate, Kitasamycin, Levofloxacin, Levofuraltadone, Levopropylcillin potassium, Lexithromycin, Lincomycin, Lincomycin hydrochloride, Linezolid, Lomefloxacin, Lomefloxacin hydrochloride, Lomefloxacin mesylate, Loraka Lubef, Mafenide, Meclocycline, Meclocycline sulfosalicylate, Megalomycin potassium phosphate, Mequidox, Meropenem, Methacycline, Methacycline hydrochloride, Methenamine, Methenamine hippurate, Methenamine mandelate, Methicillin sodium, Methioprim, Metronidazole hydrochloride, Metronidazole phosphate, Mezlocillin, Mezlocillin sodium, Minocycline hydrochloride, Mirincamycin hydrochloride Hydrochloride), monensin, monensin sodium, moxifloxacin hydrochloride, nafcillin sodium, nalidixic acid sodium, nalidixic acid, natamycin, nebramycin, neomycin palmitate, neomycin sulfate, neomycin undecylenate, netilmicin sulfate, neuthramycin, nifuraden, nifuraldeson, nifuratel, nifuratron, nifurdazil, nifurimide, nifurpirinol, nifurquinazole, nifurthiazole, nitrocycline, nitrofurantoin, nitromide, norfloxacin, novobiocin sodium, ofloxacin, ormetoprim, oxacillin sodium, oximona , oximonam sodium, oxolinic acid, oxytetracycline, oxytetracycline calcium, oxytetracycline hydrochloride, paldimycin, 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, pentizidone sodium, phenyl aminosalicylate, piperacillin, piperacillin sodium, pirbenicillinnicillin sodium, pyridicillin sodium, pirlimycin hydrochloride, pivampicillin hydrochloride, pivampicillin pamoate, pivampicillin probenate Probenate, Polymyxin B Sulfate, Porfiromycin, Propicacin, Pyrazinamide, Zinc Pyrithione, Quindecamine Acetate, Quinupristin, Racephenicol, Ramoplanin, Ranimycin, Reromycin, Repromycin, Rifabutin, Rifametane, Rifamexil, Rifamide, Rifampin, Rifapentine, Rifamixin, Rolitetracycline, Rolitetracycline Nitrate, Rosaramycin, Rosaramycin Butyrate, Rosaramycin Propionate, Rosaramycin Sodium Phosphate, Rosaramycin Stearate, Rosoxacin, Roxarsone, Roxithromycin, Sancycline, Sanfetrinem Sodium Sodium, Salmoxicillin, Sarpicillin, Scopafungin, Sisomicin, Sisomicin Sulfate, Sparfloxacin, Spectinomycin Hydrochloride, Spiramycin, Stallimycin Hydrochloride, Steffimycin, Sterile Ticarcillin Disodium, Streptomycin Sulfate, Streptonicozide, Sulbactam Sodium, Sulfabenz, Sulfabenzamide, Sulfacetamide, Sulfacetamide Sodium, Sulfacytine, Sulfadiazine, Sulfadiazine Sodium, Sulfadoxine, Sulfalene, Sulfamerazine, Sulfamethazine, Sulfamethizole, Sulfamethoxazole, Sulfamonomethoxine, Sulfamoxole, Zinc SulfanilateZinc, sulfanitran, sulfasalazine, sulfasomizole, sulfathiazole, sulfazamet, sulfisoxazole, sulfisoxazole acetyl, sulfisoxazole diolamine, sulfomixin, sulopenem, sultamicillin, sancillin sodium Sodium), talampicillin hydrochloride, tazobactam, teicoplanin, temafloxacin hydrochloride, temocillin, tetracycline, tetracycline hydrochloride, tetracycline phosphate complex, tetroxoprim, thiamphenicol, tifencillin potassium, ticarcillin credyl sodium, ticarcillin disodium, ticarcillin monosodium, ticlaton, thiodonium chloride, tobramycin, tobramycin sulfate, tosufloxacin, trimethoprim, trimethoprim sulfate, trisulfapyrimidine, troleandomycin, troleandomycin sulfate, trovafloxacin, tyrothricin, vancomycin, vancomycin hydrochloride, virginiamycin, and zorbamycin.

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

[0049] Antiviral agents useful in the present invention include, but are not limited to, immunoglobulins, amantadine, interferons, nucleotide analogs, and protease inhibitors. Specific examples of antivirals include acetomannan; acyclovir; acyclovir. Salicylic acid sodium; Adefovir; Alovudine; Alvircept Sudotox; Amantadine hydrochloride; Aranotin; Ariludone; Atevirdine mesylate; Avridine; Cidofovir; Cipamfylline; Cytarabine hydrochloride; Delavirdine mesylate; Desciclovir; Didanosine; Disoxalil; Edoxudine; Enviraden; Enviroxime; Famciclovir; Famotine hydrochloride; Fiacitabine; Fialuridine; Fosarilate; Foscarnet sodium; Phosphonate sodium; Ganciclovir; Ganciclovir sodium; Ido These include, but are not limited to, oxuridine; kethoxal; lamivudine; lobucavir; memotin hydrochloride; methisazone; nevirapine; penciclovir; pirodavir; ribavirin; rimantadine hydrochloride; saquinavir mesylate; somantadine hydrochloride; sorivudine; Statolon; stavudine; tilorone hydrochloride; trifluridine; valacyclovir hydrochloride; vidarabine; vidarabine phosphate; vidarabine sodium phosphate; viloxime; zalcitabine; zidovudine; and vinviroxime.

[0050] Nucleotide analogs are synthetic compounds that resemble nucleotides but have incomplete or abnormal deoxyribose or ribose groups. Once these nucleotide analogs enter cells, they are phosphorylated to form triphosphate forms that compete with normal nucleotides for incorporation into viral DNA or RNA. When the triphosphate forms of these nucleotide analogs are incorporated into growing nucleic acid chains, they irreversibly associate with viral polymerase, thus causing chain termination. Nucleotide analogs include, but are not limited to, acyclovir (used to treat herpes simplex virus and varicella-zoster virus), ganciclovir (useful for treating cytomegalovirus), idoxuridine, ribavirin (useful for treating respiratory syncytial virus), dideoxyinosine, dideoxycytidine, zidovudine (azidothymidine), imiquimod, and resiquimod.

[0051] Antifungal agents are used to treat superficial fungal infections as well as opportunistic and primary systemic fungal infections. Antifungal agents are useful for treating and preventing infectious fungi. Antifungal agents are sometimes classified based on their mechanism of action. Some antifungal agents function as cell wall inhibitors, for example, by inhibiting glucose synthase. These include, but are not limited to, basiungin / ECB. Other antifungal agents function by destabilizing membrane integrity. These include, but are not limited to, imidazoles such as clotrimazole, sertaconazole, fluconazole, itraconazole, ketoconazole, miconazole, and voriconazole, as well as FK463, amphotericin B, BAY38-9502, MK991, pradimicin, UK292, butenafine, and terbinafine. Other antifungal agents work by destroying chitin (e.g., chitinase) or immunosuppressing (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, furazolidone, glucocorticoids, halfantrine, iodoquinol, ivermectin, mebendazole, mefloquine, meglumine antimoniate, melarsoprol, metrifonate, metronidazole, niclosamide, nifurtimox, oxamniquine, paramomycin, pentamidine isethionate, piperazine, praziquantel, primaquine phosphate, proguanil, pyrantel pamoate, pyramidine phosphate ... These include, but are not limited to, limethamine-sulfonamide, pyrimethamine-sulfadoxine, quinacrine HCl, quinine sulfate, quinidine gluconate, spiramycin, sodium stibogluconate (sodium antimonyl gluconate), suramin, tetracycline, doxycycline, thiabendazole, tinidazole, trimethroprim-sulfamethoxazole, and triparthamide, some of which are used alone or in combination with others.

[0053] subject As used herein, a subject may be a vertebrate, including, but not limited to, a human, a mouse, a rat, a guinea pig, a rabbit, a cow, a dog, a cat, a horse, a goat, and a primate, such as a monkey. In certain aspects of the present invention, the subject may 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 and human treatment regimens. In some embodiments of the present invention, the subject is a human. In some embodiments of the present invention, the subject has a microbial infection and is in need of treatment.

[0054] In some embodiments, the subject already has or has had a microbial infection. In some embodiments, the subject is at high risk of having an infection because the subject has one or more risk factors that contribute to 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, newborns, premature babies, etc. The degree of risk of acquiring a microbial infection depends on the number and severity or importance of the risk factors the subject has. Risk charts and prediction algorithms are available to assess a subject's risk of microbial infection based on the presence and severity of risk factors. Other methods for assessing the risk of infection in a subject are known by those skilled in the art.

[0055] As used herein with respect to a subject suffering from a microbial infection, the term "infected" refers to the day the subject is infected with a microbial infectious agent, such as, but not limited to, a bacterial agent, a viral agent, a fungal agent, or a parasitic agent. It will be understood that the day of a subject's known or potential exposure to a microorganism can be considered day zero of the subject's infection with 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 infected subject's breath, saliva, droplets, exudates, bodily fluids, or excretions. 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, fabric, paper, metal, plastic, cardboard, fluids, air systems, etc. These and other means of exposure to microbial infection are known in the art.

[0056] Evaluation and Control Microbial infection in a subject can be detected using methods known in the art, including, but not limited to, assessing one or more characteristics of the microbial infection, such as the presence of a microorganism in a biological sample obtained from the subject; the level or amount of a microorganism in a biological sample obtained from the subject; and the presence and / or level of one or more physiological symptoms of the microbial infection detected in the subject. The characteristic of the microbial infection detected in the subject can be compared to a control value for the characteristic of the microbial infection. The control value can be a predetermined value that can take various forms. The control value can be a single cutoff value, such as a median or mean 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. Comparison Group Another example of the predetermined value may be a group of subjects with one or more symptoms or a diagnosis of a microbial infection and a group of subjects without one or more symptoms or a diagnosis of a microbial infection. Of course, the predetermined value will vary depending on the particular population selected. For example, a group of individuals with a microbial infection who are administered a gelsolin agent and not an antibacterial agent may have one or more different characteristics of the microbial infection than a group of individuals with a microbial infection who are administered an antibacterial agent and not a gelsolin agent. Thus, the selected predetermined value can take into account the category into which the individual falls. An appropriate category can be selected by one of ordinary skill in the art through simple routine experimentation.

[0057] Controls can be used in the methods of the present invention to compare characteristics of different control groups, characteristics of a subject with characteristics of a control group, etc. Comparisons between subjects and controls, comparisons between one control and another, etc., can be based on relative differences. For example, and not by way of limitation, the physiological symptoms of a subject treated with a synergistic therapeutic method of the present invention, comprising administering to the subject a gelsolin agent and an antimicrobial agent, can be compared to the physiological symptoms of a control group to which the gelsolin agent is administered but not the antimicrobial agent. Comparisons can be expressed in relative terms; for example, if an increase in body temperature (indicating a fever) or a decrease in body temperature is characteristic of a microbial infection, the body temperature of a subject treated with a synergistic therapeutic method of the present invention can be compared to the body temperature level of a control. In some embodiments, a suitable control is a subject not treated with a synergistic therapeutic method of the present invention. Comparisons between a treated subject and a control can include comparing the percentage difference in temperature between the treated subject and the selected control. In some cases, the body temperature of a subject treated with the methods of the present invention can be determined to be lower compared to a selected control, where the subject's body temperature is lower by 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%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 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%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8 %, 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.

[0058] In certain instances, the body temperature of a subject treated with the methods of the present invention can be determined to be elevated compared to a selected control, where the subject's body temperature is elevated by 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%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 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%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8 %, 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.

[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 with a microbial infection. The results of the assay in a subject treated using the synergistic treatment method of the present invention can be compared with the level of microbial infection in a control, e.g., the results of the assay for 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 present invention can be compared with the results of the assay in a subject treated using the synergistic treatment method of the present invention. The results can be compared to a control to determine the percentage difference between the subject and control levels. In some embodiments, the level of microbial infection in treated subjects is less than 100% of the control infection level. In certain embodiments of the invention, the level of microbial infection in a treated subject 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%, 52%, 53%, 54%, 55%, 55%, 56%, 57%, 57%, 58%, 58%, 59 ... 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.

[0060] In another non-limiting example, the increase in the level of microbial infection and / or the therapeutic effect of an antimicrobial agent using the methods of the present invention can be determined by comparing the viability of a control to the viability of a subject treated with a synergistic method or composition of the present invention. A non-limiting example of control viability is the viability of a subject with a microbial infection that is not treated with a method of the present invention. Non-limiting examples of viability parameters that can be measured include determining the length of time (hours, days, weeks, etc.) a subject remains alive after treatment with the present invention, and determining whether the subject dies or survives after treatment with the present invention. It will be understood how to compare these and other parameters related to viability with a control to evaluate and determine the therapeutic effect of a synergistic method or composition of the present invention. A non-limiting example of control viability is the number of days a subject survives after treatment with a 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 each of the antimicrobial agent and gelsolin agent. In some embodiments of the invention, the survival chance of a subject treated with a synergistic method of the 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%, or 500% greater than the survival chance of a control.

[0061] In another non-limiting example, the increase in the level of microbial infection and / or the therapeutic effect of an antimicrobial agent using the method of the present invention can be determined by comparing the level of lung lesions in a control with the level of lung lesions in a subject treated with a synergistic method or composition of the present invention. A non-limiting example of the level of lung lesions in a control is the level of lung lesions in a subject with a microbial infection who has not been treated with the method of the present invention. Non-limiting examples of measurable lung lesion parameters include determining the lung histopathology in a subject. In a non-limiting example, the histopathology of lung tissue (e.g., 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 in a blinded manner by a board-certified pathologist. A scoring system can be used to compare the subject's lung tissue with that of a control. In a non-limiting example, lung lesions can be evaluated using a 4-point, 4-criteria system (inflammation; infiltration; necrosis; and other, including hemorrhage) for a maximum score of 16 points. Points for each criterion can be assigned based on pathology findings: absent (0), minimal (1), mild (2), moderate (3), and severe (4). A scoring system can be used to assess lung pathology by comparing target tissue with control tissue. Additional means of comparing lung pathology are known in the art and are described in the present invention. In some embodiments of the invention, subjects treated with a synergistic method of the invention have a level of lung pathology that 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%, or 500% lower than the level of lung pathology in a control.

[0062] In another non-limiting example, the level of microbial infection and / or the increased therapeutic effect of an antimicrobial agent using the methods of the present invention can be determined by comparing the level of weight loss or relative weight loss of a control with the level of weight loss or relative weight loss of a subject treated with a synergistic method or composition of the present invention. A non-limiting example of a control level of weight loss is the level of weight loss in a subject with a microbial infection not treated with a method of the present invention. Non-limiting examples of weight loss and / or relative weight loss parameters that can be measured include the subject's weight before the microbial infection, the subject's weight during the microbial infection before treatment with a synergistic method of the present invention, the subject's weight after receiving a synergistic treatment method of the present invention, etc. In a non-limiting example, the weight of a subject with a Pseudomonas aeruginosa infection can be determined before and after administration of a synergistic treatment of the present invention comprising a gelsolin agent and a carbapenem class agent, a non-limiting example of which is meropenem. The subject's weight can be compared to the subject's pre-treatment weight, pre-infection weight, and / or the weight of another control. A decrease in weight loss in a subject after administration of a synergistic treatment of the present invention indicates a reduction in the microbial infection in the subject. In some embodiments of the invention, the level of weight loss in a subject treated with a synergistic method of the 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%, or 500% less than the level of weight loss in a control.

[0063] It will be understood that the control can be a predetermined value as well as a sample of material that is tested in parallel with the experimental material.Examples include a sample from a control population or a control sample produced through manufacturing that is tested in parallel with the experimental sample;A control can also be a sample from a subject before, during, or after treatment with an embodiment of the method or composition of the present invention.Therefore, one or more characteristics determined for a subject with infection can later be used as the "control" value of those characteristics in that subject.

[0064] In some embodiments of the invention, the effectiveness of the synergistic methods of the invention can be assessed by comparing the synergistic therapeutic results in subjects treated using the methods of the invention with one or both of the following: (1) the individual therapeutic effects of the gelsolin agent and (2) the individual therapeutic effects of the antibacterial agent. In certain aspects of the invention, the difference in the level of therapeutic effectiveness can be assessed on a scale indicating the increase from the control level. In some aspects, the increase is from the control zero level obtained in (1) or (2) to a level greater than zero resulting from treatment with the synergistic methods of the invention. In some embodiments of the invention, the level of therapeutic effect of the synergistic treatment method of the invention is increased 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 therapeutic effect of a control.

[0065] Delayed medication method Some embodiments of the present invention include a delayed dosing schedule determined to be effective in reducing viral infection in an infected subject. Delaying administration of the gelsolin agent to the subject until three or more days after the day the subject contracts a viral infection (day zero) enhances the therapeutic effect of the gelsolin agent. Some embodiments of the treatment methods of the present invention include administering an effective amount of a gelsolin agent to a subject with a viral infection, wherein the gelsolin agent is administered to the subject with a viral infection at least three, four, five, six, seven, eight, nine, or more days after the subject's infection. In some embodiments, the gelsolin agent is not administered to the subject on the day the subject contracts a viral infection (day zero). In some embodiments, the gelsolin agent is not administered on the first day (day 1) after the subject contracts a viral infection. In some embodiments, the gelsolin agent is not administered on the second day (day 2) after the subject contracts a viral infection. In some embodiments of the methods of the present invention, the gelsolin agent is not administered on one or more of days 0, 1, and 2 of the subject's viral infection.

[0066] As used herein with respect to a subject suffering from a microbial infection, the term "infected" refers to the day on which the subject becomes infected with a microbial infectious agent, such as, but not limited to, a bacterial agent, a viral agent, a fungal agent, etc. It will be understood that the day of the subject's known or potential exposure to a microbial agent can be considered 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 the subject's known or potential exposure to the virus; (2) administering an antiviral agent to a subject within 48 hours of the subject's known or potential exposure to the virus; (3) seasonal prophylaxis using an antiviral agent by administering an antiviral agent to a subject without specific known exposure to the virus; and (4) prophylaxis using an antiviral agent in the context of a community outbreak of the virus. Exposure to a viral infection is understood to mean direct or indirect contact with an individual suffering from a viral infection. Non-limiting examples of contact with an infected individual include physical contact, i.e., contact with the breath, saliva, droplets, exudates, bodily fluids, excretions, etc., of an infected individual. In some embodiments, indirect contact can be physical contact by a subject with a substrate contaminated by an infected individual. Examples of substrates that may be contaminated by an individual suffering from a viral infection include, but are not limited to, food, fabric, paper, metal, plastic, cardboard, fluids, air systems, etc. These and other means of exposure to viral infections 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] The method for treating a viral infection using a timed / delayed gelsolin agent dosing regimen can include administering a gelsolin agent at a determined time delay following a subject's known exposure to a viral infection, suspected exposure to a viral infection, potential exposure to a viral infection, and / or risk of exposure to a viral infection. The administered gelsolin agent can include a gelsolin molecule, a functional fragment thereof, or a functional derivative of a gelsolin molecule. In some embodiments, the gelsolin molecule is plasma gelsolin (pGSN), and in certain embodiments of the methods of the present invention, the gelsolin molecule is a recombinant gelsolin molecule.

[0069] In some embodiments, an effective amount of gelsolin agent is administered to a subject in a manner that is comparable to the therapeutic effect of a control. and increasing the therapeutic effect of the administered gelsolin agent against a viral infection in the subject, wherein the control therapeutic effect comprises the therapeutic effect that would occur if the gelsolin agent were not administered to the subject. 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% greater than the therapeutic effect of the control.

[0070] In certain embodiments of the present invention, the therapeutic effect of administering a gelsolin agent reduces the level of viral infection in a subject compared to a control level of viral infection, which can be the level of infection in the absence of administration of a gelsolin agent. In some embodiments of the present invention, the level of viral infection in a subject after administration of a gelsolin agent in the methods 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 subject can be determined by, for example, using one or more of the following methods: detecting the presence, absence and / or level of viral infection characteristics in biological samples obtained from subject; observing subject; assessing one or more physiological symptoms of viral infection in subject; assessing the viability of subject; or other methods known in the art.Physiological symptoms of viral infection include, but are not limited to, one or more of fever, fatigue, weight loss and death.

[0072] Embodiments of the present invention may include administering an effective amount of a gelsolin agent to a subject 3, 4, 5, 6, 7, or more days after the subject's exposure or suspected exposure to a viral infection, wherein administration of the effective amount of the gelsolin agent increases the subject's viability compared to a control viability, where the control viability is the viability in the absence of administration of the gelsolin agent. The increase in subject viability after administration of the gelsolin agent using a 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% greater than the control viability.

[0073] A time delay in administering a gelsolin agent to a subject up to three or more days after the day the subject contracts a viral infection (day zero) enhances the therapeutic effect of the gelsolin agent, and the time delay can be used in combination with administration of an antiviral agent, thereby resulting in a synergistic effect between the antiviral agent and the gelsolin agent administered to the subject. In some aspects of the present invention, the methods of treating a viral infection of the present invention include administering an antiviral agent to the subject one or more days before the time-delayed administration of a gelsolin agent to the subject. In some embodiments, the antiviral agent can be administered before the subject's exposure or potential exposure to a viral infection, or on day 0, 1, or 2 of the subject's exposure or suspected exposure to a viral infection. It has been determined that effective amounts of each of the gelsolin agent and antiviral agent administered to a subject can have a synergistic therapeutic effect against a viral infection compared to the therapeutic effect of a control in which both the gelsolin agent and the antiviral agent are not administered to the subject in a synergistic manner. As described elsewhere herein, the antiviral agent is administered in a clinically tolerated amount, and it is understood that the control therapeutic effect can be the therapeutic effect of administration of a clinically tolerated amount of the antiviral agent administered without administration of the gelsolin agent.

[0074] In some embodiments of the methods of the invention, the clinically tolerated amount of the antiviral agent is an amount that is less than the maximum tolerated dose (MTD) of the antiviral agent. D is the highest possible, yet tolerable dose level of the antiviral agent for the subject.In certain instances, the MTD of the antiviral agent is determined at least in part based on the preselected clinical limiting toxicity of the antiviral agent.In the methods of the present invention, comprising administering synergistically effective amounts of a gelsolin agent and an antiviral agent, the synergistic effect reduces the minimum effective dose (MED) of the antiviral agent in the subject.In certain methods of the present invention, the MED is the lowest dose level of the antiviral agent that provides a clinically significant response in mean efficacy, and this response is statistically significantly greater than the response provided by a control that does not contain this dose of 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 antivirals: oseltamivir phosphate (available in generic versions or under the trade name Tamiflu®), zanamivir (trade name Relenza®), and peramivir (trade name Rapivab®); and cap-dependent endonuclease (CEN) inhibitors: for example, baloxavir marboxil (trade name Xofluza®).

[0076] Antiviral therapies for preventing and treating viral infections, such as influenza A, B, C, and D infections, are known and routinely used in the art. It is also recognized that certain viral strains may be resistant to known antiviral therapies (see, for example, Moscona, A., 20090, 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 antiviral agents. 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 antiviral agents.

[0077] Certain embodiments of the methods of the present invention treat viral infections using a timed-dose gelsolin regimen administered in the absence of an antiviral regimen. Some embodiments of the methods of the present invention treat viral infections by administering an antiviral regimen and a time-delayed gelsolin regimen to a subject in need of such treatment. In some embodiments of the methods of the present invention, administering an antiviral agent and a delayed-dose gelsolin agent to a subject results in a synergistic therapeutic effect of the gelsolin 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 compared to the therapeutic effect of a control. 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 compared to the therapeutic effect of a control.

[0078] Drug preparation and administration The methods and compositions of the present invention have important implications for patient treatment and the clinical development of new therapeutic methods. Clinical researchers are also expected to use the methods of the present invention to determine the entry criteria for human subjects in clinical trials. Medical professionals select therapeutic regimens for treatment based on the expected net benefit to the subject. Net benefit is derived from the risk-to-benefit ratio.

[0079] The amount of treatment can be modified, for example, by increasing or decreasing the amount of gelsolin agent and / or antimicrobial agent administered to the subject, by changing the therapeutic composition administered, by changing the route of administration, by changing the timing of administration, etc. The effective amount will vary depending on factors within the knowledge and expertise of a medical practitioner, such as the particular infection or condition being treated, the age and physiological condition of the subject being treated, the severity of the infection or condition, the duration of treatment, the particular route of administration, etc. For example, the effective amount may depend on the extent to which an individual has been exposed to or affected by exposure to a microbial infection.

[0080] Effective dose The term "effective amount" as used herein in connection with the treatment methods or compositions of the present invention is referred to as a "synergistically effective amount." The methods of the present invention include administering a gelsolin agent and an antibacterial agent in amounts that are synergistically effective amounts of the gelsolin agent and the antibacterial agent, respectively. When administered to a subject using the methods of the present invention, the synergistically effective amounts of the gelsolin agent and the antibacterial agent result in a synergistic therapeutic effect against and / or a reduction in microbial infection in the subject.

[0081] An effective amount is the dosage of each drug sufficient to provide medically desirable results. 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, gelsolin and antibacterial agents. It should be understood that the drugs of the present invention are used to treat or prevent infection, i.e., they can be used prophylactically in subjects at risk of developing infection. Thus, an effective amount is an amount that can reduce, delay, or in some cases completely prevent the risk of developing infection. It is recognized that when a drug is used in an acute situation, it is used to prevent one or more medically undesirable consequences that usually result from such adverse events.

[0082] The factors involved in determining effective dose are well known to those skilled in the art, and can be handled by routine experimentation alone.Usually preferred to use the maximum dose of the medicament of the present invention (alone or in combination with other therapeutic agents), that is, the highest safe dose according to sound medical judgment.However, those skilled in the art will understand that patients may insist on a lower dose or tolerated dose for medical reasons, psychological reasons, or other reasons in fact.

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

[0084] Drugs and Delivery The medicaments used in the methods of the present invention are preferably sterile and contain an effective amount of gelsolin and an effective amount of an antibacterial agent to produce the desired response in a unit of weight or volume suitable for administration to a subject. The dose of the medicament administered to a subject can be selected according to various parameters, particularly the mode of administration used and the subject's condition. Other factors include the desired duration of treatment. In the event that the subject's response is inadequate with the initial dose applied, higher doses (or effectively higher doses via a different, more localized delivery route) can be used, as tolerated by the patient. The dosage of the medicament can be adjusted by the individual physician or veterinarian, especially if complications arise. Therapeutically effective amounts typically vary from 0.01 mg / kg to about 1000 mg / kg, from about 0.1 mg / kg to about 200 mg / kg, or from about 0.2 mg / kg to about 20 mg / kg, administered in one or more daily doses for one or more days. The gelsolin and antibacterial agents may also be referred to herein as drugs.

[0085] Various modes of administration that effectively deliver the agents of the present invention to the desired tissues, cells, or body fluids are known to those skilled in the art. The method and dosage to be administered can be adjusted by individual physicians, medical professionals, or veterinarians, especially when complications occur. The absolute amount to be administered will depend on the material selected for administration, whether the administration is a single dose or multiple doses, and the age, physical condition, etc. The amount of time that the individual subject will have to live will depend on a variety of factors, including individual subject parameters, including age, size, weight, and stage of the disease or condition, which are well known to those of skill in the art and can be addressed with no more than routine experimentation.

[0086] Pharmaceutically acceptable carriers include diluents, fillers, salts, buffers, stabilizers, solubilizers, 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, and the like. While salts may be pharmaceutically acceptable when used in medicine, non-pharmaceutically acceptable salts may be conveniently used to prepare pharmaceutically acceptable salts, and this is 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, hydrobromide, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, citric acid, formic acid, malonic acid, succinic acid, and the like. Pharmaceutically acceptable salts may also be prepared as alkali metal or alkaline earth salts, such as sodium, potassium, or calcium salts.

[0087] Various administration modes known to those skilled in the art can be used to effectively deliver the pharmaceutical compositions of the present invention, including antibacterial agents and gelsolin agents, to a subject to produce a synergistic therapeutic effect against a microbial infection 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 administration. In some embodiments of the present invention, the means for administering the compositions of the present invention is inhalation. The present invention is not limited by the specific administration modes disclosed herein. Standard references in the art (e.g., Remington, The Science and Practice of Pharmacy, 2012, edited by Allen, Loyd V., Jr., 22nd Edition) provide administration modes and formulations for delivering various pharmaceutical preparations and formulations to pharmaceutical carriers. Other protocols useful for administering the therapeutic compounds of the present invention are known to those skilled in the art, and the dosages, administration schedules, administration sites, administration modes (e.g., intraorgan), etc., may differ from those presented herein. Other protocols useful for administering the agents of the present invention will be known to those skilled in the art, and will differ in dosage, administration schedule, site of administration, mode of administration, etc. from those presented herein.

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

[0089] When administered, the pharmaceutical preparations of the present invention are administered in pharmaceutically acceptable amounts and in pharmaceutically acceptable compositions. The term "pharmaceutically acceptable" refers to a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient. Such preparations may typically contain salts, buffers, preservatives, compatible carriers, and optionally other therapeutic agents. While salts must be pharmaceutically acceptable when used in medicine, non-pharmaceutically acceptable salts may conveniently be used to prepare pharmaceutically acceptable salts, and this is 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, hydrobromide, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, citric acid, formic acid, malonic acid, succinic acid, etc. Pharmaceutically acceptable salts may also be prepared as alkali metal or alkaline earth salts, such as sodium, potassium, or calcium salts.

[0090] If desired, the drug or composition can be combined with a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers, diluents or encapsulating substances suitable for human administration. The term "carrier" refers to 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 drug of the present invention and with each other in such a way that there is no interaction that would substantially impair the desired medicinal effect.

[0091] The pharmaceutical compositions may contain suitable buffering agents, including acetate, phosphate, citrate, glycine, borate, carbonate, bicarbonate, hydroxide (and other bases), and pharmaceutically acceptable salts of the foregoing compounds, as described above. The pharmaceutical compositions may also optionally contain suitable preservatives, such as benzalkonium chloride, chlorobutanol, parabens, and thimerosal.

[0092] The pharmaceutical composition can be conveniently provided in unit dosage form and can be prepared by any method known in the field of pharmacy.All methods include the step of combining the active agent with the carrier, which constitutes one or more accessory ingredients.Generally, the composition is prepared by uniformly and intimately combining the active compound with a liquid carrier, a finely divided solid carrier, or both, and then, if necessary, shaping the product.

[0093] Compositions suitable for oral administration can be presented as discrete units such as capsules, tablets, pills, lozenges, each containing a predetermined amount of the active compound (e.g., gelsolin). 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 milled, and after adding suitable additives as needed, the granular mixture can be processed to obtain tablets or dragee cores. Suitable excipients can be, 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). If desired, disintegrants, such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate, can be added. Optionally, oral preparations can be formulated in saline or buffer, i.e., EDTA to neutralize internal acid conditions, or can be administered without a carrier.

[0095] Oral dosage forms of one or more of the above components are also specifically contemplated. One or more components can be chemically modified to enable effective oral delivery of the derivatives. Generally, the contemplated chemical modification is the attachment of at least one moiety to the component molecule itself, which (a) inhibits proteolysis; and (b) allows uptake into the bloodstream from the stomach or intestine. Also desirable is an increase in the overall stability of these components and their circulation time in the body. Examples of such moieties include polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, and polyproline. Abuchowski and Davis, 1981, "Soluble Polymer-Enzyme Adducts," In: Enzymes as Drugs, edited by Hocenberg and Roberts, Wiley-Interscience, New York, NY, 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] For pharmaceuticals, the location of release may be the stomach, the small intestine (the duodenum, jejunum, or ileum), or the large intestine. Those skilled in the art have available formulations that do not dissolve in the stomach but release the substance in the duodenum or elsewhere in the intestine. Preferably, the release avoids the harmful effects of the stomach environment, either by protection of the gelsolin agent and / or antimicrobial agent, or by release of the biologically active substance beyond the stomach 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 should be in dosages suitable for such administration.

[0098] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner. For administration by inhalation, the compound for use according to the present invention can be conveniently provided in the form of aerosol spray from pressurized pack or nebulizer by using suitable propellant, for example, dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoromethane, carbon dioxide or other suitable gas.In the case of pressurized aerosol, dosage unit can be determined by providing a valve for delivering a metered amount.For example, the capsules and cartridges of gelatin for use in inhaler or insufflator can be formulated to contain the powder mixture of compound and suitable powder base, for example, lactose or starch.

[0099] Pulmonary delivery of gelsolin is also contemplated herein. Gelsolin is delivered to the lungs of a mammal during inhalation and crosses the pulmonary epithelial lining to the bloodstream. Nasal (or intranasal) delivery of the pharmaceutical composition of the present invention is also contemplated.Nasal delivery allows the pharmaceutical composition of the present invention to pass into the bloodstream immediately after administering the therapeutic product to the nose, without the need for the product to be deposited in the lungs.Formulations for nasal delivery include formulations containing dextran or cyclodextran.

[0100] When it is desired to deliver the compound systemically, it can be formulated for parenteral administration by injection, for example, by bolus injection or continuous infusion.The preparation for injection can be presented in a unit dosage form, for example, in an ampoule or a multi-dose container with added preservative.The composition can take the form of suspension, solution or emulsion in oily or aqueous vehicle, and can contain compounding agents such as suspending agent, stabilizer and / or dispersing agent.

[0101] Preparations for parenteral administration include aqueous solutions of water-soluble active compounds.In addition, suspensions of active compounds 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 can contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran.Optionally, the suspension can also contain suitable stabilizers or agents that increase the solubility of the compound, allowing for the preparation of highly concentrated solutions.Alternatively, the active compound can be in powder form, which can be reconstituted with a suitable vehicle, such as sterile pyrogen-free water, before use.

[0102] In particular, agents including but not limited to gelsolin and antimicrobial agents may be provided in particles. As used herein, particles refer to any or all of the gelsolin or antimicrobial agents described herein. "Nanoparticles" refers to nanoparticles or microparticles (or possibly larger) that may be composed, in part, of a nanoparticle or microparticle. The particle may contain a drug within a core surrounded by a coating, including but not limited to an enteric coating. The drug may also be dispersed throughout the particle. The drug may also be adsorbed onto the particle. The particle may have 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. In addition to the drug, the particle may contain any of the materials routinely used in the pharmaceutical and medical fields, including but not limited to erodible, nonerodible, biodegradable, or nonbiodegradable materials or combinations thereof. The particle may be a microcapsule containing gelsolin in solution or in a semi-solid state. The particle may be of virtually any shape.

[0103] Both non-biodegradable and biodegradable polymeric materials can be used to manufacture particles for drug delivery. Such polymers can be natural or synthetic. The polymer is selected based on the desired release period. 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, glutin, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isopropyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate).

[0104] The drug may be contained 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 called "sustained release") is used in its conventional sense to refer to a formulation that provides gradual release of drug over an extended period of time, preferably, but not necessarily, resulting in a substantially constant blood concentration of drug over an extended period of time. The term "delayed release" is used in its conventional sense to refer to a drug formulation in which there is a time delay between administration of the formulation and the release of the drug therefrom. "Delayed release" may or may not involve gradual release of drug over an extended period of time, and therefore may or may not be a "sustained release."

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

[0106] The present invention also contemplates the use of kits. In some embodiments of the present invention, the kits can include one or more pharmaceutical preparation vials, pharmaceutical preparation diluent vials, an antimicrobial agent, and a gelsolin agent. The vial containing the diluent for the pharmaceutical preparation is optional. The diluent vial can include a diluent such as saline for diluting what may be a concentrated solution or lyophilized powder of the gelsolin agent and / or antimicrobial agent. The instructions can include instructions for mixing a specific amount of diluent with a specific amount of the concentrated pharmaceutical preparation, thereby preparing a final formulation for injection or infusion. The instructions can also include instructions for treating a subject with an effective amount of the gelsolin agent and the antimicrobial agent. The container containing the preparation, whether it is a bottle, a vial with a septum, an ampoule with a septum, an infusion bag, or the like, can be used to automate the preparation. It is also understood that the container may include indicia such as conventional indicia that change color when claved or otherwise sterilized.

[0107] The present invention is further illustrated by the following examples, which should not be construed as further limiting in any way. The entire contents of all references cited throughout this application (including literature references, issued patents, published patent applications, and co-pending patent applications) are 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 invention. As will be apparent to those skilled in the art, the present invention will find use in a variety of compositions and methods. [Example]

[0109] Example 1 Antibiotic-resistant pneumococcal pneumonia is a potential problem. Research has been conducted to evaluate novel therapeutic strategies to combat infection, including measures to enhance innate immunity. Experiments were performed to determine the effects of pGSN administration on macrophages and host survival. method: Bacterial species and cultures S. pneumoniae serotype 3 (catalog no. 6303, American Type Culture Collection, Rockville, MD) was cultured overnight on 5% sheep blood-supplemented agar Petri dishes (catalog no. 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 studies In vitro studies were performed in which 125–250 μg / ml of pGSN was added to bacterial cultures to determine bacterial viability. (2) In vivo studies B16 mice were insufflated for 10 min. 5 Mice were challenged with 10 mg of pneumococcus pneumoniae and given 10 mg of pGSN sc 2 hours before, 8 hours after, and 20 hours after infection. In some studies, pGSN was administered as an aerosol 15 or 30 minutes before infection. Aerosols were generated using a 5 mg / ml solution as in Hamada, K. et al., J. Immunology. 2003;170(4):1683-9.

[0110] Results / Discussion 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 sc 2 h before and 8 and 20 h after infection, 10 mg sc by insufflation) significantly improved macrophage uptake (Figure 1B). 5 pGSN improved bacterial clearance in Bl6 mice challenged with pneumococci (fewer viable bacteria at 24 hours) (Fig. 1C); similar results were seen when pGSN was administered as an aerosol 15 or 30 minutes before infection; aerosols were generated using a 5 mg / ml solution as in Hamada, K., et al., J. Immunology. 2003;170(4):1683-9 (Fig. 1D). Systemic pGSN (sc) prevented primary pneumococcus (Fig. 1E, 3 × 10 5 CFU inoculation) or secondary post-influenza pneumococcal pneumonia (Figure 1F, using a 500 CFU inoculation on day 7 after mild influenza infection with PR8). *=p <.05 vs. control, n = 6–12 per group. Serotype 3 Strep. Pneumoniae [ATCC #6303] was used in all experiments.

[0111] Macrophage NOS3 is a key 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). Our results indicated that this pathway functions as a key mechanism of pGSN effects on macrophages, as pGSN failed to improve bactericidal responses 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 antibiotic-susceptible and antibiotic-resistant mouse models of pneumococcal pneumonia. method: 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). Serotype 3 bacteria were grown 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 we followed the growth protocol reported in Restrepo AV et al., BMC Microbiol 2005;5:34, which uses two sequential growths in liquid broth cultures prior to centrifugation and adjustment of 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). Primary pneumococcal pneumonia was induced as previously reported (Yang Z. et al., Am J Physiol Lung Cell Mol Physiol 2015;309:L11-6). For antibiotic-susceptible pneumonia, 1.5-2 × 10 Streptococcus pneumoniae type 3 was inoculated under ketamine (72 mg / kg i.p.) plus xylazine (9.6 mg / kg i.p.) anesthesia. 6 Mice were instilled intranasally with colony-forming units (CFU). 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, distance measurement experiments revealed approximately 300 × 10 colony-forming units (CFU) were used for instillation under anesthesia as described above. 6 A highly lethal inoculum of colony-forming units (CFU) was identified. Most studies used 10 mice per group in vehicle, penicillin (PEN), pGSN, or PEN+pGSN groups.

[0115] Treatment and Outcomes Recombinant human pGSN (rhu-pGSN) was synthesized in E. coli and purchased from Fujifilm Purified by Diosynth (Billingham, UK). rhu-pGSN was administered to mice by intraperitoneal injection at doses ranging from 5 to 10 mg, as detailed in the Results section. In some experiments, penicillin (G Procaine Injectable Suspension, NDC) was used. Mice were administered 0.1–2 mg of fluoxetine (57319-485-05, Phoenix Pharmaceuticals) intramuscularly via intramuscular injection. Mice were monitored for 10 days and survival, weight change, and overall morbidity were measured using a composite index adapted from the guidelines of Burkholder T. et al., Curr Protoc Mouse Biol 2012;2:145–65 (i.e., hunched appearance, ruffled fur, or partially closed eyes each scored 1 point; protruding penis or splayed hindquarters scored 1.5 points; lethargy scored 2 points; maximum score 8; evaluation was performed unblinded to treatment group). For animals that did not survive, weight and morbidity scores from the last day of survival were carried forward. To assess lung inflammation by quantifying neutrophil influx, one cohort of animals underwent lung lavage 48 hours after euthanasia and infection as previously described (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, resuspended lavage fluid samples were counted by hemocytometer, and differential cell counts were performed on Wright-Giemsa-stained cytocentrifuge preparations.

[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 measures, differences between groups were examined by ANOVA.

[0117] result Delayed treatment with rhu-pGSN was tested in the same mouse model previously used to demonstrate improved 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 post-infection with serotype 3 pneumococci significantly improved survival from a highly lethal inoculum compared to vehicle controls, even in the absence of antibiotic treatment. In contrast to subsequent experiments using serotype 14, 100% survival in antibiotic-treated mice confirmed that serotype 3 is highly susceptible to penicillin (Figure 3B).

[0118] To determine whether these findings extend to antibiotic-resistant pneumonia, we developed a similar model using the highly virulent serotype 14 pneumococcus. Treatment began 24 hours postinfection 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 resistance of this strain in vitro (Jabes D. et al., J Infect Dis 1989;159:16-25).

[0119] During the 24 hours prior to treatment, all mice experienced identical deterioration, as evidenced by comparable weight loss and morbidity scores. Neutrophil influx at 48 hours postinfection was reduced in animals treated with a single dose of pGSN with or without penicillin (total washed 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). rhu-pGSN treatment alone resulted in significant improvement in overall survival, recovery from weight loss, and improved morbidity scores (Figure 4A–C).

[0120] In vitro, penicillin treatment alone or in combination with pGSN did not affect bacterial growth (increase in bacterial CFU, 1-hour (h) incubation with vehicle, PEN (16 μg / ml) or PEN + pGSN (250 μg / ml): 88,000, 105,000, and 88,000, respectively, average of two replicates).

[0121] In vivo, treatment with the combination of penicillin and pGSN resulted in higher survival rates than 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 nine experiments, survival was highest in the pGSN + PEN group, followed by pGSN alone compared to either PEN or vehicle alone (the pGSN + PEN combination was significantly better than pGSN alone). The table in Figure 5 provides results from nine experiments in which four delayed-administration treatments were evaluated. Data from the final four experiments, which used essentially identical treatments and are representative of the overall results obtained across all nine studies, are shown in Figures 4A-C. Figure 5 provides details of all nine experiments, including the pilot and distance measurement studies. Column H shows the change in bacterial growth method for the superior growth results obtained using the 2-fold growth method in BHI broth (Restrepo AV et al., BMC Microbiol 2005;5:34) for penicillin-resistant Streptococcus pneumoniae. The difference in viability was statistically significant as determined by analysis of all nine pooled studies using log-rank analysis with Sidak correction for multiple comparisons. The results of the statistical analysis of the final four experiments (numbers 6-9) are detailed in Figure 4A-C.

[0122] Consideration The study was designed and constructed to mimic the clinical situation that subjects exhibit after infection becomes apparent. Therefore, the experiment was performed using a clinically relevant scenario in which administration was delayed until mice were visibly ill, rather than the pre-treatment or co-treatment used in previous studies (Yang Z. et al., Am J Physiol Lung Cell Mol Physiol 2015;309:L11-6). This design was used to evaluate the potential of pGSN to improve treatment outcomes. A key finding was that delayed pGSN treatment improved survival when used alone without antibiotics or in combination with suboptimal antibiotics to which the bacterial strain was highly resistant. The reduced bronchoalveolar neutrophil counts observed in infected pGSN-treated animals may reflect accelerated bacterial clearance due to pGSN-stimulated resident macrophages, the inflammation-modulating activity of pGSN, or both. For serotype 14, our ability to study longer delays before treatment was limited in our pilot study by the relatively high number of deaths by the second or third day without treatment. We will conduct further studies to investigate other antibiotic-resistant bacteria in other model systems. Previous findings that pGSN enhances macrophage bactericidal function 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 totality of the data suggests a synergistic interaction between pGSN and penicillin treatment, which was ineffective by itself. However, this conclusion relies on the analysis of all pooled distance measurements, not just the final test performed. When analyzing only the final four replicate studies (Figure 4A-C), the comparison is in the same direction but does not achieve statistical significance. No enhancement of penicillin effects on bacterial growth in vitro by concomitant rhu-pGSN was observed. While not intending to be bound by any particular theory, these data suggest that antibacterial defenses enhanced by pGSN may be even more effective against bacteria that are slightly perturbed (but not killed) by penicillin. This mechanism This mechanism merits future attention, especially if similar results are observed in other infections caused by resistant bacteria. Collectively, rhu-pGSN can improve outcomes in a highly lethal pneumococcal pneumonia model when given after a clinically appropriate delay, even in the setting of antimicrobial resistance. These findings support further evaluation of pGSN as an adjunctive therapy for severe antibiotic-resistant infections.

[0124] Example 3 Highly lethal multidrug-resistant Pseudomonas in a neutropenic mouse model A study was conducted to evaluate the efficacy of rhu-pGSN treatment with meropenem in aeruginosa pneumonia. method Generating rhu-pGSN Recombinant human plasma gelsolin (rhu-pGSN) was produced in E. coli and then lyophilized for reconstitution. A vehicle control containing the formulation components was used for comparison mice. Bacterial species and growth conditions Pseudomonas aeruginosa UNC-D was isolated from sputum from a patient with cystic fibrosis [Lawrenz MB et al., Pathog. Dis. 73 (2015)]. Bacteria were grown on trypticase soy agar (TSA) plates and in Lennox broth at 37°C with shaking broth cultures. The minimum inhibitory concentrations for UNC-D strains 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]. Bacteria were grown overnight in Lennox broth, washed with 1x PBS, and then analyzed at OD . 600 It was prepared for animal challenge studies by diluting to a final concentration based on the base estimate and a final 50 μl delivery volume. The bacterial inoculum was confirmed by serial dilution and colony counting on TSA plates.

[0125] Animal respiratory infection models The BALB / c infection model with Pseudomonas aeruginosa UNC-D strain [Lawrenz MB et al. (2015) Pathog. Dis. 73(5):ftv025] was specifically designed to test adjuvant therapies that could improve the efficacy of failed meropenem monotherapy against multidrug-resistant (MDR) Pseudomonas aeruginosa UNC-D strains resistant to several clinically important antibiotics, including meropenem. Previous experience has demonstrated that this model is most informative when investigating novel compounds using a meropenem dose that provides approximately 50% mortality with meropenem treatment alone [Lawrenz MB et al. (2015) Pathog. Dis. 73(5):ftv025]. Mice were housed and treated in accordance with standard animal experimentation guidelines at the University of Louisville. Briefly, female BALB / c mice were made neutropenic using cyclophosphamide injections (150 mg / kg) on days −5 and −3 before infection, which typically resulted in a reduction of neutrophil counts by approximately 90%. 5.5 CFU of UNC-D were instilled directly into the lungs by intratracheal instillation via intubation. Meropenem (Hospira; Lake Forest, IL) was administered by subcutaneous injection every 8 hours for 5 days, starting 3 hours after infection.

[0126] To determine whether adjunctive rhu-pGSN therapy improved 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 postinfection. Mice were monitored for disease development every 8 hours for 7 days postinfection via transponders (BioMedic Data Systems; Seaford, DE) implanted subcutaneously before the start of the study. Temperatures measured at the time of the test were also included. Moribund mice were humanely euthanized and scored as having succumbed to infection at the following time points: Tissue samples were collected for bacterial counts and pathology as previously described [Lawrenz MB et al., Pathog. Dis. 73 (2015)]. Mice surviving up to 7 days were scored as having survived infection and euthanized; tissues were processed similarly. Lung histopathology was scored in a blinded manner by a board-certified veterinary pathologist. Lung lesions were assessed using a 4-point, 4-criterion system (inflammation; infiltration; necrosis; and other, including hemorrhage) for a maximum score of 16 points. Points for each criterion were assigned based on pathology findings: absent (0), minimal (1), mild (2), moderate (3), and severe (4).

[0127] statistical analysis In total, three parallel experiments were performed independently using this model. Titration experiments were performed using new batches of meropenem to determine the effective dose (ED) of each lot of antibiotic prior to the formal experiment. 50 The data were used to estimate survival rates. Overall survival and survival with minimal lung damage (post-hoc defined as a histopathology score ≤2) were tabulated for the entire experiment and for experimental conditions in which the meropenem-only control group protected ≤50% of mice. The 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 a normal approximation to the binomial distribution. For individual experimental conditions in which the mortality rate in the control meropenem group was approximately 50% or greater, survival curves were analyzed by the log-rank test, temperature data by two-way ANOVA, and bacterial load and pathology scores by one-way ANOVA with Tukey's post-hoc multiplicity adjustment. The pre-specified primary endpoint was survival rate 7 days after infection challenge. During the analysis of these data, a "survival-plus" endpoint examining survival with healthy lungs (histopathology score ≤2) was used as a clinically meaningful extension of a favorable outcome. Bacterial load and temperature response were not included in this two-pronged composite because they are not direct measures of clinical improvement.

[0128] result rhu-pGSN improved survival in mice infected with Pseudomonas aeruginosa To determine whether rhu-pGSN can improve the efficacy of meropenem against pulmonary infection, female BALB / c mice were made neutropenic with cyclophosphamide (n=8), infected with MDR Pseudomonas aeruginosa, and treated with various doses of meropenem to determine the time at which meropenem therapy begins to fail in this model (i.e., the ED of meropenem). 50 A dose approaching 500 mg / kg / day was determined. Mice were treated with selected doses of meropenem with or without rhu-pGSN for 5 days postinfection and monitored for the development of moribund disease for 7 days postinfection (Figure 6). In both experiments 1 and 2, treatment with 1250 mg / kg / day of meropenem resulted in a survival rate of ≤50%, indicating failure of meropenem treatment and allowing confirmation of whether adjunctive therapy with rhu-pGSN could improve efficacy. Focusing on animals receiving this dose, the addition of rhu-pGSN numerically increased the number of animals surviving to the end of each study (Figure 7A-B). Combining the two consecutive studies, 31% of mice receiving meropenem alone survived for 7 days, compared to 75% survival when mice were given meropenem 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 in the meropenem-only group), no difference in survival was observed between treatment groups (Figure 6).

[0129] To determine whether the increased survival rate with rhu-pGSN therapy was associated with a reduced lung bacterial burden, lung samples were collected from the lungs of mice receiving 1250 mg / kg / day at the time of euthanasia. Colony counts were determined (Figure 8A-C). A general trend was observed suggesting that rhu-pGSN improved control of bacterial burden in the lungs of infected mice compared to meropenem alone, although a statistically significant difference in bacterial counts was only observed in the second study (p=0.0273).

[0130] Overall survival across all treatment groups in the three combined experiments was 35 / 64 (55%) and 46 / 64 (72%) in mice treated with meropenem without or with rhu-pGSN, respectively [Δ(95% confidence interval) = 17% (1, 34)]. Adjunctive rhu-pGSN treatment enhanced the efficacy of meropenem against pulmonary infection with Pseudomonas aeruginosa, although inhibition of bacterial growth in the lungs may only partially explain the observed effect. Interestingly, while meropenem alone controlled spread from the lung to the spleen in both studies, pGSN enabled splenic colonization in some animals. This observation was not significant alone in any study, but combined data demonstrated a significant increase in spleen counts in pGSN-treated mice. These observations, along with improved survival, were consistent with rhu-pGSN exerting an opsonizing effect to enhance splenic uptake.

[0131] rhu-pGSN limits acute lung injury The lack of a clear relationship between reduced lung bacterial burden and increased survival in mice receiving rhu-pGSN raised the possibility that rhu-pGSN protection was mediated by alternative or additional mechanisms. Because pGSN modulates inflammation, we investigated the question of whether lung injury was reduced by rhu-pGSN adjunctive therapy in animals infected with Pseudomonas aeruginosa receiving 1250 mg / kg / day of pGSN. Representative sections of lung tissue collected from animals were blindly scored for pathology by a board-certified veterinary pathologist. The addition of rhu-pGSN to meropenem reduced host lung injury (Figure 9A-B; p = 0.0035 and p = 0.1514, respectively). Combining the data from these two independent studies, the mean pathology score for mice receiving meropenem alone was 6.86, whereas the mean pathology score for mice receiving both meropenem and rhu-pGSN was 2.53 (Figure 9C; p = 0.0049).

[0132] Based on these observations that rhu-pGSN conferred protection from lung injury, we expanded our analysis to include mice receiving doses of meropenem above and below 1250 mg / kg / day. Overall survival of mice receiving different doses of meropenem in three individual experiments is shown in Figure 6. Animals that survived infection for 7 days were grouped as those exhibiting either near-normal lung histology (pathology score ≤ 2) or signs of lung pathology (pathology score > 2). Retrospectively, using this criterion, overall survival with mild lung injury was observed in 26 / 64 (41%) mice receiving meropenem alone and 38 / 64 (59%) mice given meropenem plus rhu-pGSN [Δ(95% confidence interval) = 19% (2, 36)] (Figure 10). To eliminate noise generated by highly effective and ineffective meropenem doses, arbitrary but clinically reasonable exclusion limits of ≥ 75% and ≤ 25% were imposed on control survival. At this midpoint of response to meropenem alone, another exploratory post-hoc analysis showed that 12 / 32 (37.5%) patients had a favorable outcome (survival with near-normal lungs) with meropenem alone and 27 / 32 (84.4%) with meropenem plus rhu-pGSN [Δ=47% (26,68)].

[0133] Using surviving mice as the denominator, near-normal lung histopathology was observed in 26 / 35 (74.3%) and 38 / 46 (82.6%) mice treated with meropenem monotherapy versus meropenem and rhu-pGSN combination therapy, respectively. These data combined suggest that the addition of rhu-pGSN significantly reduced the risk of Pseudomonas aeruginosa infection compared with mice treated with antibacterial agents alone. It has been shown that steroids can reduce lung damage caused by P. uginosa infection.

[0134] Plasma gelsolin accelerates the resolution of the host's systemic response As part of monitoring disease progression, host body temperature was tracked over the course of infection. In this model, all mice tended to exhibit a steady decline in body temperature within the first 24 hours after infection. In mice receiving effective treatment, temperature eventually returned to normal, while temperatures continued to decline in mice receiving less effective treatment [Lawrenz MB et al. (2015) Pathog.Dis.73(5):ftv025]. The temperature-normalized time course allowed for assessment of differences in recovery rates between different treatments. In these experiments, the target ED of meropenem alone was significantly higher than that of meropenem alone. 50 Focusing on dosing regimens approaching rhu-pGSN, we investigated the question of whether pGSN promoted the recovery of temperature homeostasis in mice that survived infection. In the two studies that achieved a survival advantage, mice typically experienced a drop in body temperature of approximately -12.22°C (10°F) within the first 24 hours after infection (Figure 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, host body temperature recovery was much more rapid in mice treated with rhu-pGSN and meropenem, with survivors' temperatures recovering to 35°C (95°F) by day 2. Thus, adjunctive rhu-pGSN not only improved survival and lung lesions but also accelerated systemic recovery in the host, as measured by temperature curves. In a third experiment in which no survival benefit was observed with rhu-pGSN, no differences in temperature course were observed between treatment groups.

[0135] Consideration The addition of rhu-pGSN to meropenem improved survival in an established murine model of severe, multidrug-resistant Pseudomonas aeruginosa pneumonia. Temperature normalization in surviving mice generally occurred more rapidly with rhu-pGSN adjuvant therapy than with meropenem alone. Lungs from rhu-pGSN recipients generally contained fewer viable bacteria. Furthermore, rhu-pGSN reduced the degree 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 rhu-pGSN-mediated reductions in bacterial burden and the severity of lung injury during the course of infection.

[0136] The first line of host defense against infection involves a focused inflammatory response. However, excessive local and systemic inflammation can be detrimental to vital organs proximal and distal to the primary infection site. Once acute injury recedes, pGSNs promote the resolution of the inflammatory process and limit the resulting damage.

[0137] The potential effect of adding rhu-pGSN treatment to meropenem was examined in a highly lethal, multidrug-resistant Pseudomonas aeruginosa pneumonia model in a neutropenic mouse model. All mice died within approximately 24 hours of infection without immediate antimicrobial therapy. rhu-pGSN as a sole treatment extended mean survival by approximately 12 hours. Titration experiments were performed with each batch of antibiotic to determine the meropenem dose that resulted in ≥50% mortality. Nevertheless, outcomes were not always predictable, with some trials resulting in ≤25% or ≥75% mortality compared with the meropenem control. Under these extreme conditions, mice were either too sick or not sufficiently sick, potentially masking the potential effect of adjunctive rhu-pGSN on outcomes. Nevertheless, rhu-pGSN given with meropenem was more effective than meropenem alone under most conditions.

[0138] These preclinical data support the development of rhu-pGSN as an adjunct to standard treatment modalities. This adds to the growing body of evidence that rhu-pGSN may be effective in limiting lung injury while increasing survival. No serious or drug-related adverse events were observed in rhu-pGSN recipients treated for 3 consecutive days, even at supraphysiological levels throughout the dosing interval.

[0139] Using an established model of murine Gram-negative pneumonia, bacterial colony counts from alveolar lavage and histopathological lung injury scores at euthanasia were higher in mice receiving meropenem alone compared with mice treated with meropenem and rhu-pGSN, although considerable intra- and interexperimental variability was observed. Most notably, adding rhu-pGSN to meropenem reduced both mortality and parenchymal injury, most notably in situations where meropenem alone was relatively ineffective.

[0140] Example 4 method Mouse models 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 before the start of the experiment. Separate batches of mice were used for each study. A 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 with an intraperitoneal injection of 72 mg / kg ketamine plus 9.6 mg / kg xylazine. Mice then received an intranasal instillation of 25 μl of a PBS suspension containing virus (ranging from 400 to 1000 PFU depending on the study) or vehicle alone. All infections were performed at approximately the same time (approximately 10:00 AM). Initial titration identified 400 PFU as the dose resulting in approximately 60% mortality in vehicle-treated mice, and this dose was used in the majority of studies (see Figure 12). Most studies used at least 10 mice per group for vehicle and pGSN treatment groups; details of influenza doses, total number of mice, and their weights are provided in the underlying data table [Kobzik L: "Expanded Tables 1 & 2". Harvard Dataverse, V1 2019. www.doi.org / 10.7910 / DVN / 53GJY1].

[0141] Treatment and Outcomes Recombinant human pGSN (rhu-pGSN) was synthesized in E. coli and purchased from Fujifilm Purified rhu-pGSN was obtained from Diosynth (Billingham, UK). Based on previous demonstration of its function in rodent models and because data using human gelsolin would facilitate clinical translation efforts, we used human gelsolin rather than mouse gelsolin. Rhu-pGSN was administered to mice daily by subcutaneous injection starting on days 3 or 6 postinfection at doses ranging from 0.5 to 5 mg, as detailed in the Results section. Mice were monitored for 12 days and survival, weight change, and overall morbidity were measured using a composite index adapted from previously described guidelines [Burkholder T et al., Current Protocols Mouse Biol. 2012;2:145-65.] (i.e., hunched appearance, ruffled fur, or partially closed eyes each scored 1 point; protruding penis or splayed hindquarters scored 1.5 points; lethargy scored 2 points; maximum score 8; evaluations were performed unblinded to treatment groups). For animals that did not survive, the weight and morbidity scores from the last day of survival were carried forward.

[0142] Lung transcriptome profiling from mice treated with either vehicle or rhu-pGSN at 7 days postinfection and Lung tissue was obtained on day 9 (2 mg per day was administered starting on day 3 postinfection, 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 uses highly multiplexed RT-PCR amplification and next-generation sequencing (NGS) quantification to measure the expression of 4753 protein-encoding functionally important mouse genes. Amplification index libraries were generated and sequenced on an Illumina NextSeq 500 instrument according to the procedures detailed in section 5.3 of the Cellecta user manual. Sequencing data were converted to FASTQ format for further analysis using DriverMap Sample Extraction software. This produced a raw data matrix file of counts for each sample in 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. A Breslow-Day test for homogeneity of pGSN vs. vehicle comparisons across studies yielded p>0.2, indicating that homogeneity was not rejected and supporting comparisons across studies performed via a log-rank (Mantel-Cox) test stratified by test. For other measures, differences between groups were investigated by ANOVA. Transcriptome profiling results, scaled to normalize column counts, were converted 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 allow evaluation using a custom background gene list (i.e., approximately 4700 genes measured using the Cellecta DriverMap platform).

[0144] result Effect of rhu-pGSN on survival To evaluate the potential for rhu-pGSN to improve outcomes, various dose and timing regimens were tested, with a total of 18 studies performed, tabulated in Figure 12 and summarized in Figure 13. To mimic potential clinical use, mice were not treated until several days after challenge.

[0145] The main finding was that delayed treatment with rhu-pGSN resulted in a significant improvement in mouse survival (Figures 14A-H). Combining all studies, 39% (93 / 236) surviving mice treated with vehicle and 62% (241 / 389) surviving mice treated with pGSN were observed at day 12 (p=0.000001, Figure 14A). Improved survival was observed regardless of whether delayed treatment was initiated at day 6 (Figure 14C) or day 3 (Figures 14E, 14G) post-infection. Similarly, rhu-pGSN reduced morbidity scores compared to vehicle treatment (Figures 14B, 14D, 14F, 14H). In contrast, no statistically significant differences in weight loss or recovery (surviving animals) were consistently observed in the experiments summarized in Figures 14A-H. The only exception was found in a study that initially tested a low-dose regimen (>2 mg rhu-pGSN on days 3–6 / 7, then 5 mg until day 11). In this latter series of studies, body weights at the end of the study (compared to day 0) were 81.4 ± 4.7% in vehicle-treated mice, whereas pGSN and 85±2.6% in treated mice (p<0.0001; see summary of 4 studies, also Figures 12 and 13, and more detailed tables of all experiments in Expanded Data [Kobzik L: "Expanded Tables 1 & 2". Harvard Dataverse, V1 2019. www.doi.org / 10.7910 / DVN / 53GJY1]). The beneficial effect of rhu-pGSN was observed in most, but not all, of the 18 individual studies (Figure 12, see Discussion).

[0146] Transcriptome profiling To assess whether rhu-pGSN treatment altered the transcriptome profile of infected lungs [see Harvard Dataverse: Expanded Tables 1 & 2. https: / / doi.org / 10.7910 / DVN / 53GJY116 ], lung tissue was collected (n = 5 per group per day) just before (day 7) and just after (day 9) the onset of normal mortality in this model (day 8). Per protocol, the dose of rhu-pGSN was 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 down-regulated genes and 149 up-regulated genes. The top 50 up- and down-regulated genes are shown in Figure 15, which is notable for the many cytokine and immune-related genes prominent among the down-regulated genes in the rhu-pGSN-treated group (e.g., IL10, IL12rb, CTLA4, and CCR9, 7, and 5, among others). Gene enrichment analysis of the complete down-regulated gene list was performed using the Panther online analysis tool, querying the GO Ontology or Reactome databases. The primary finding was a decreased expression of biological processes related to immune and inflammatory responses, or the release of cytokines and other cellular activators. The top 10 most significant processes / pathways are shown in Figure 16. Analysis using another gene enrichment analysis software tool (MetaCore) produced similar results. Analysis of the up-regulated gene list identified enrichment for processes related to tissue morphogenesis and epithelial / epidermal cell differentiation (consistent with repair of influenza-mediated damage; see Discussion).Details of the DriverMap gene list, the identified differentially expressed genes, and the complete results of the gene enrichment analysis, which uses the down- and up-regulated gene lists to query the Panther and MetaCore databases, are presented in worksheets 2–15 in the spreadsheets available in Expanded Data [Kobzik L: Harvard Dataverse, V1 2019. www.doi.org / 10.7910 / DVN / 8HBFD7]. Data for the experimental groups of the study described herein are presented in Tables 1 and 2 of Harvard Dataverse: Expanded Tables 1 & 2. http: / / doi.org / 10.7910 / DVN / 53GJY116, where additional variables such as weights and statistical analyses are also described. Additional data from the experiments described herein are provided in 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] Consideration We performed a study to evaluate the potential of rhu-pGSN to improve outcomes in severe influenza using a clinically relevant scenario of delayed treatment initiation. The key finding was that delayed pGSN treatment significantly improved survival even when used from day 3 or up to day 6 after infection. The study was conducted immediately after the patient's infection (immediately after the onset of severe symptoms). Given the impracticality of initiating early treatment (as opposed to initiating early treatment), as well as adverse outcomes observed in some experimental models, the delay was implemented so as not to interfere with the immediate immune response to influenza.

[0148] Some limitations merit consideration. First, there is the experimental variability observed. Treatment with rhu-pGSN increased survival in the majority of experiments performed, but not all of them. Some negative trials were attributed to factors including, but not limited to, technical issues with virus stocks, variations in instillation methods, and insufficient initial rhu-pGSN doses in the "low-dose, then high-dose" trials. Wherever possible, methods were adjusted to reduce these potential sources of variability.

[0149] Experimental variables were also manipulated to examine, for example, whether treatment as late as day 6 versus day 3 after the start of infection was effective, and other variables in the study were evaluated. Ultimately, beneficial effects were observed whether the survival analysis included all trials (Figures 14A, B) or trials using treatment from day 6 or day 3 (Figures 14C-H).

[0150] When surviving mice were euthanized, they were followed for only 12 days. Because the survival curve was still potentially declining, final mortality could not be determined with certainty. However, the time to death was at least prolonged with rhu-pGSN compared with placebo treatment.

[0151] Notably, although rhu-pGSN did not rescue all mice that died from influenza in the experimental model, the results demonstrated a significant survival benefit. Given the goal of identifying novel treatments for severe influenza, the results obtained in mice without supportive fluids, additional therapeutic agents (e.g., but not limited to, antivirals), and respiratory care provided to hospitalized patients support the conclusion that the method will provide similar and synergistic benefits in clinical settings. The results suggest that administering gelsolin at an appropriate time after infection, combined with standard treatments such as antivirals, may provide a greater survival benefit.

[0152] In summary, rhu-pGSN, when given after a clinically relevant delay, can improve outcomes in a highly lethal murine influenza model. 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 pathogen type. Experimental results support the use of gelsolin as an adjunctive therapy for severe influenza and other viral infections in humans and other mammals.

[0153] Example 5 Further studies are carried out using synergistic amounts of gelsolin and antiviral agents. In certain studies, oseltamivir phosphate, zanamivir, peramivir, or baloxavir marboxil is the antiviral drug administered to the subject. The gelsolin is administered using a delayed dose method as described herein above. Effective amounts of the antiviral and gelsolin are administered to a subject with or suspected of having a viral infection, such as influenza A, B, C, or D, and the effective amounts produce a synergistic therapeutic effect against the virus in the subject's infection. A synergistic therapeutic effect is a greater improvement in one or more characteristics of the viral infection in the subject than the improvement in one or more characteristics in a control, and the control does not receive treatment including the administration of a synergistically effective amount of gelsolin and antiviral agents.

[0154] equivalent Although several embodiments of the present invention have been described and illustrated herein, those skilled in the art will recognize that the present invention is not limited to the embodiments described herein. Various other means and / or structures for performing the functions and / or obtaining the results and / or advantages of one or more of the inventions described herein are readily envisioned, and each such variation and / or modification is deemed to be within the scope of the present invention. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are intended to be exemplary, and that the 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 recognize, or be able to ascertain using no more than 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 invention may be practiced otherwise than as specifically described and claimed. The present invention relates to each individual feature, system, article, material, and / or method described herein. Furthermore, any combination of two or more of such features, systems, articles, and / or methods, where such features, systems, articles, and / or methods are not mutually inconsistent, is included within the scope of the present invention.

[0155] All definitions defined and used herein should be understood to cover dictionary definitions, definitions in documents incorporated by reference, and / or ordinary meanings of the defined terms.

[0156] The indefinite articles "a" and "an," as used in the specification and claims, unless clearly indicated to the contrary, should be understood to mean "at least one." The term "and / or," as used in the specification and claims, should be understood to mean "either or both" of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Unless expressly indicated to the contrary, other elements may optionally be present other than those specifically identified by the "and / or" clause, whether associated with specifically identified elements or not.

[0157] All references, patents and patent applications and publications cited or referred to in this application are incorporated herein by reference in their entirety.

Claims

1. A composition comprising an effective amount of a gelsolin agent and an antibacterial agent for synergistically treating a microbial infection in a subject.

2. The composition of claim 1, wherein the antibacterial agent is in a clinically acceptable amount and the administered gelsolin agent and antibacterial agent synergistically enhance the therapeutic effect of administering a clinically acceptable amount of the antibacterial agent to a subject without administering the gelsolin agent.

3. 2. The composition of claim 1, wherein the clinically tolerated amount of the antibacterial agent is an amount that is less than the maximum tolerated dose (MTD) of the antibacterial agent in the subject.

4. 10. The composition of claim 1, wherein the MTD of the antibacterial agent is the highest possible, yet tolerable, dose level of the antibacterial agent to a subject.

5. 5. The composition of claim 4, wherein the MTD of the antibacterial agent is determined at least in part based on a preselected clinically limiting toxicity for the antibacterial agent in the subject.

6. 10. The composition of claim 1, wherein the synergistically effective amounts of the gelsolin agent and the antibacterial agent reduce the minimal effective dose (MED) of the antibacterial agent in the subject.

7. 7. The composition of claim 6, wherein the MED is the lowest dose level of the antibacterial agent that provides a clinically significant response in mean efficacy, which response is statistically significantly greater than the response provided by a control that does not include that dose of the antibacterial agent.

8. The composition of claim 1 , wherein the synergistic therapeutic effect of the gelsolin agent and the antibacterial agent comprises increasing the survival chances of the subject.

9. 10. The composition of claim 1, wherein the synergistic therapeutic effect of the gelsolin agent and the antibacterial agent comprises reducing a microbial infection in a subject.

10. 2. The composition of claim 1, wherein the microbial infection is a bacterial infection, optionally caused by a Pneumococcus species.

11. The composition of claim 1, wherein the antibacterial agent comprises a β-lactam antibiotic.

12. The composition of claim 1 , wherein the antibacterial agent comprises penicillin.

13. 10. The composition of claim 1, wherein the microbial infection is caused by a type of Pseudomonas aeruginosa.

14. 10. The composition of claim 1, wherein the antibacterial agent is a carbapenem class antibacterial agent.

15. 2. The composition of claim 1, wherein the antibacterial agent is meropenem.

16. The composition of claim 1 , wherein the antibacterial agent comprises an antifungal agent and the microbial infection comprises a fungal infection.

17. The composition of claim 1 , wherein the antimicrobial agent comprises an antiparasitic agent and the microbial infection comprises a parasitic infection.

18. The composition of claim 1 , wherein the antibacterial agent comprises an antiviral agent and the microbial infection comprises a viral infection.

19. The composition of claim 1 , wherein the subject is a mammal, optionally a human.

20. 10. The composition of claim 1, wherein the gelsolin agent comprises plasma gelsolin (pGSN), optionally recombinant pGSN.

21. The composition of claim 1 further comprising a pharmaceutically acceptable carrier.

22. The composition of claim 1 , wherein the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of a gelsolin molecule.

23. The composition of claim 1 further comprising a pharmaceutically acceptable carrier.

24. 1. A method for increasing the therapeutic efficacy of an antimicrobial agent against a microbial infection in a subject, comprising: A method comprising administering synergistically effective amounts of a gelsolin agent and an antibacterial agent to a subject having a microbial infection, wherein the administered gelsolin agent and antibacterial agent have a synergistic therapeutic effect on the subject's microbial infection, the synergistic therapeutic effect being greater than the therapeutic effect of the antibacterial agent administered without the gelsolin agent.

25. 25. The method of claim 24, wherein the antibacterial agent is administered in a clinically tolerated amount.

26. The method of claim 24, wherein the synergistic therapeutic effect against the microbial infection is greater 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.

27. 27. The method of claim 26, wherein the therapeutic effect of the control is equal to the individual therapeutic effect of the gelsolin agent.

28. 27. The method of claim 26, wherein the control therapeutic effect is equal to the individual therapeutic effect of the antibacterial agent administered in a clinically tolerated amount.

29. 28. The method of claim 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% greater than the therapeutic effect of the control.

30. 25. The method of claim 24, wherein the antimicrobial agent comprises an antibiotic agent and the microbial infection comprises a bacterial infection.

31. 25. The method of claim 24, wherein the antibacterial agent comprises an antifungal agent and the microbial infection comprises a fungal infection.

32. 25. The method of claim 24, wherein the antimicrobial agent comprises an antiparasitic agent and the microbial infection comprises a parasitic infection.

33. 25. The method of claim 24, wherein the antibacterial agent comprises an antiviral agent and the microbial infection comprises a viral infection.

34. 25. The method of claim 24, wherein the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of a gelsolin molecule.

35. 35. The method of claim 34, wherein the gelsolin molecule is plasma gelsolin (pGSN).

36. 36. The method of claim 34 or 35, wherein the gelsolin molecule is a recombinant gelsolin molecule.

37. 26. The method of claim 25, wherein the clinically tolerated amount of the antibacterial agent is an amount that is less than the maximum tolerated dose (MTD) of the antibacterial agent.

38. 38. The method of claim 37, wherein the MTD of an antibacterial agent is the highest possible, yet tolerable, dose level of the antibacterial agent for a subject.

39. 39. The method of claim 38, wherein the MTD of the antimicrobial agent is determined based at least in part on a preselected clinically limiting toxicity for the antimicrobial agent.

40. 25. The method of claim 24, wherein the synergistically effective amounts of the gelsolin agent and the antimicrobial agent reduce the minimal effective dose (MED) of the antimicrobial agent in the subject.

41. 25. The method of claim 24, wherein the synergistic therapeutic effect of administering a synergistically effective amount of each of the antibacterial agent and the gelsolin agent reduces the level of microbial infection in the subject compared to the level of microbial infection in a control.

42. 42. The method of claim 41, wherein the control infection level comprises the infection level in the absence of administration of a synergistically effective amount of each of the antibacterial agent and the gelsolin agent.

43. 43. The method of claim 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 microbial infection in the control.

44. 42. The method of claim 41, wherein the level of microbial infection in the subject is determined, and the means of determining comprises one or more of an assay, observing the subject, assessing one or more physiological symptoms of microbial infection in the subject, and assessing the viability of the subject.

45. 45. The method of claim 44, wherein the physiological symptoms include one or more of fever, malaise, and mortality.

46. 45. The method of claim 44, wherein the physiological condition comprises a pulmonary lesion.

47. 45. The method of claim 44, wherein the physiological symptoms include weight loss.

48. 45. The method of claim 44, wherein the assay comprises a means for detecting the presence, absence, and / or level of a characteristic of a microbial infection in a biological sample from a subject.

49. 25. The method of claim 24, wherein administration of a synergistically effective amount of each of the antimicrobial agent and the gelsolin agent increases the survival chance of the subject compared to the survival chance of a control.

50. 50. The method of claim 49, wherein the control viability is the viability in the absence of administration of a synergistically effective amount of each of the antimicrobial agent and the gelsolin agent.

51. 51. The method of claim 49 or 50, wherein the increase in survival chance 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% greater than survival chance of the control.

52. 25. The method of claim 24, wherein administration of a synergistically effective amount of each of the antibacterial agent and the gelsolin agent reduces the level of lung lesions in the subject compared to the level of lung lesions in a control.

53. 53. The method of claim 52, wherein the control level of lung lesions is the level of lung lesions in the absence of administration of a synergistically effective amount of each of the antibacterial agent and the gelsolin agent.

54. The method of claim 52 or 53, wherein the level of lung lesions in a subject administered synergistically effective amounts of each of an antibacterial agent and a gelsolin 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 a control.

55. 53. The method of claim 52, wherein the subject has a Pseudomonas aeruginosa bacterial infection.

56. 53. The method of claim 52, wherein the antibacterial agent comprises the carbapenem class, optionally including meropenem.

57. 31. The method of claim 30, wherein the bacterial infection is caused by a type of Streptococcus pneumoniae (pneumococcus).

58. 31. The method of claim 30, wherein the antibacterial agent comprises a β-lactam antibiotic.

59. 31. The method of claim 30, wherein the antibacterial agent comprises penicillin.

60. 31. The method of claim 30, wherein the bacterial infection is caused by a type of Pseudomonas aeruginosa.

61. 61. The method of claim 60, wherein the antibacterial agent is an antibacterial agent of the carbapenem class.

62. 62. The method of claim 61, wherein the antibacterial agent is meropenem.

63. 31. The method of claim 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.

64. 25. The method of claim 24, wherein 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, intrasynovial, and intraocular administration.

65. 25. The method of claim 24, wherein the subject is a mammal, optionally a human.

66. 25. The method of claim 24, wherein the gelsolin agent is a non-therapeutic gelsolin agent.

67. 25. The method of claim 24, wherein the antimicrobial agent is a non-therapeutic agent.

68. A method for synergistically treating a microbial infection in a subject, comprising administering to a subject having a microbial infection effective amounts of a gelsolin agent and an antibacterial agent, respectively, wherein the administered gelsolin agent and antibacterial agent have a synergistic therapeutic effect on the subject's microbial infection compared to the therapeutic effect of a control, and the antibacterial agent is administered in a clinically acceptable amount.

69. 69. The method of claim 68, wherein the control comprises the therapeutic effect of administration of a clinically tolerated amount of the antibacterial agent administered without administration of the gelsolin agent.

70. 69. The method of claim 68, wherein the clinically tolerated amount of the antibacterial agent is an amount that is less than the maximum tolerated dose (MTD) of the antibacterial agent.

71. 71. The method of claim 70, wherein the MTD of the antibacterial agent is the highest possible, yet tolerable, dose level of the antibacterial agent for the subject.

72. 72. The method of claim 71, wherein the MTD of the antimicrobial agent is determined based at least in part on a preselected clinically limiting toxicity for the antimicrobial agent.

73. 69. The method of claim 68, wherein the synergistically effective amounts of the gelsolin agent and the antibacterial agent reduce the minimal effective dose (MED) of the antibacterial agent in the subject.

74. 74. The method of claim 73, wherein the MED is the lowest dose level of the antibacterial agent that provides a clinically significant response in mean efficacy, which response is statistically significantly greater than the response provided by a control that does not include that dose of the antibacterial agent.

75. 69. The method of claim 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% greater than the therapeutic effect of the control.

76. 69. The method of claim 68, wherein the antimicrobial agent comprises an antibiotic agent and the microbial infection comprises a bacterial infection.

77. 69. The method of claim 68, wherein the antibacterial agent comprises an antifungal agent and the microbial infection comprises a fungal infection.

78. 69. The method of claim 68, wherein the antimicrobial agent comprises an antiparasitic agent and the microbial infection comprises a parasitic infection.

79. 69. The method of claim 68, wherein the antibacterial agent comprises an antiviral agent and the microbial infection comprises a viral infection.

80. The gelsolin agent is a gelsolin molecule, a functional fragment thereof, or a 69. The method of claim 68, comprising a functional derivative.

81. 69. The method of claim 68, wherein the gelsolin molecule is plasma gelsolin (pGSN).

82. 82. The method of claim 80 or 81, wherein the gelsolin molecule is a recombinant gelsolin molecule.

83. 69. The method of claim 68, wherein the synergistic therapeutic effect of administering a synergistically effective amount of each of the antibacterial agent and the gelsolin agent reduces the level of microbial infection in the subject compared to the level of microbial infection in a control.

84. 84. The method of claim 83, wherein the control infection level comprises the infection level in the absence of administration of a synergistically effective amount of each of the antibacterial agent and the gelsolin agent.

85. 84. The method of claim 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 microbial infection in the control.

86. 84. The method of claim 83, wherein the level of microbial infection in the subject is determined, and the means of determining comprises one or more of an assay, observing the subject, assessing one or more physiological symptoms of microbial infection in the subject, and assessing the viability of the subject.

87. 87. The method of claim 86, wherein the physiological symptoms include one or more of fever, malaise, and death.

88. 87. The method of claim 86, wherein the physiological symptoms include weight loss.

89. 87. The method of claim 86, wherein the physiological condition comprises a pulmonary lesion.

90. 87. The method of claim 86, wherein the assay comprises a means for detecting the presence, absence, and / or level of a characteristic of a microbial infection in a biological sample from a subject.

91. 69. The method of claim 68, wherein administering a synergistically effective amount of each of the antibacterial agent and the gelsolin agent increases the survival chance of the subject compared to the survival chance of a control.

92. 92. The method of claim 91, wherein the control viability is the viability in the absence of administration of a synergistically effective amount of each of the antimicrobial agent and the gelsolin agent.

93. 92. The method of claim 91, wherein the increase in subject survival probability 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% greater than the survival probability of the control.

94. 69. The method of claim 68, wherein administration of a synergistically effective amount of each of the antibacterial agent and the gelsolin agent reduces the level of lung lesions in the subject compared to the level of lung lesions in a control.

95. 95. The method of claim 94, wherein the control level of lung lesions is the level of lung lesions in the absence of administration of a synergistically effective amount of each of the antibacterial agent and the gelsolin agent.

96. The method of claim 94, wherein the level of lung lesions in a subject administered synergistically effective amounts of each of an antibacterial agent and a gelsolin 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 a control.

97. 77. The method of claim 76, wherein the subject has a Pseudomonas aeruginosa bacterial infection.

98. 95. The method of claim 94, wherein the antibacterial agent comprises the carbapenem class, optionally including meropenem.

99. 77. The method of claim 76, wherein the bacterial infection is caused by a type of Streptococcus pneumoniae (pneumococcus).

100. 77. The method of claim 76, wherein the antibacterial agent comprises a beta-lactam antibiotic.

101. 77. The method of claim 76, wherein the antibacterial agent comprises penicillin.

102. 77. The method of claim 76, 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.

103. 69. The method of claim 68, wherein 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, intrasynovial, and intraocular administration.

104. 69. The method of claim 68, wherein the subject is a mammal.

105. 69. The method of claim 68, wherein the gelsolin agent is a non-therapeutic gelsolin agent.

106. 69. The method of claim 68, wherein the antibacterial agent is a non-therapeutic agent.

107. 1. A pharmaceutical composition comprising an antibacterial agent and a gelsolin agent that synergistically increases the therapeutic effect of the antibacterial agent against a microbial infection, for use in a method of treating a subject, comprising: The subject has a microbial infection, and the method includes administering a pharmaceutical composition comprising a synergistically effective amount of each of a gelsolin agent and an antibacterial agent in an amount effective to treat the microbial infection in the subject, wherein the synergistic therapeutic effect is greater than the therapeutic effect of the antibacterial agent administered without the gelsolin agent.

108. 108. The pharmaceutical composition of claim 107, wherein the gelsolin agent and the antibacterial agent are administered to the subject separately or simultaneously.

109. The pharmaceutical composition of claim 107, wherein the antibacterial agent is administered in a clinically tolerated amount, and the administered gelsolin agent and antibacterial agent synergistically enhance the therapeutic effect of administering a clinically tolerated amount of the antibacterial agent to a subject without administering the gelsolin agent.

110. 110. The pharmaceutical composition of claim 109, wherein the clinically tolerated amount of the antibacterial agent is an amount that is less than the maximum tolerated dose (MTD) of the antibacterial agent in a subject.

111. 111. The pharmaceutical composition of claim 110, wherein the MTD of the antibacterial agent is the highest possible, yet tolerable, dose level of the antibacterial agent for a subject.

112. 111. The pharmaceutical composition of claim 110, wherein the MTD of the antibacterial agent is determined at least in part based on a preselected clinically limiting toxicity for the antibacterial agent in the subject.

113. 108. The pharmaceutical composition of claim 107, wherein the synergistically effective amounts of the gelsolin agent and the antibacterial agent reduce the minimal effective dose (MED) of the antibacterial agent in a subject.

114. The pharmaceutical composition of claim 113, wherein the MED is the lowest dose level of the antibacterial agent that provides a clinically significant response in mean efficacy, which response is statistically significantly greater than the response provided by a control that does not include that dose of the antibacterial agent.

115. 108. The pharmaceutical composition of claim 107, wherein the synergistic therapeutic effect of the gelsolin agent and the antibacterial agent comprises increasing the survival chances of the subject.

116. 108. The pharmaceutical composition of claim 107, wherein the synergistic therapeutic effect of the gelsolin agent and the antibacterial agent comprises reducing a microbial infection in a subject.

117. 108. The pharmaceutical composition of claim 107, wherein the microbial infection is a bacterial infection, optionally caused by a Pneumococcus species.

118. 108. The pharmaceutical composition of claim 107, wherein the antibacterial agent comprises penicillin.

119. 108. The pharmaceutical composition of claim 107, wherein the bacterial infection is caused by one type of Pseudomonas aeruginosa.

120. 108. The pharmaceutical composition of claim 107, wherein the antibacterial agent is an antibacterial agent of the carbapenem class.

121. 108. The pharmaceutical composition of claim 107, wherein the antibacterial agent is meropenem.

122. 108. The pharmaceutical composition of claim 107, wherein the antibacterial agent comprises an antifungal agent and the microbial infection comprises a fungal infection.

123. 108. The pharmaceutical composition of claim 107, wherein the antibacterial agent comprises an antiparasitic agent and the microbial infection comprises a parasitic infection.

124. 108. The pharmaceutical composition of claim 107, wherein the antibacterial agent comprises an antiviral agent and the microbial infection comprises a viral infection.

125. The pharmaceutical composition of claim 107, wherein the subject is a mammal.

126. 108. The pharmaceutical composition of claim 107, wherein the gelsolin agent comprises plasma gelsolin (pGSN), optionally recombinant pGSN.

127. 108. The pharmaceutical composition of claim 107, further comprising a pharmaceutically acceptable carrier.

128. 108. The pharmaceutical composition of claim 107, wherein the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of a gelsolin molecule.

129. 108. The pharmaceutical composition of claim 107, further comprising a pharmaceutically acceptable carrier.

130. A method for treating a viral infection in a subject, comprising administering an effective amount of a gelsolin agent to a subject having a viral infection, wherein the gelsolin agent is administered at least 3, 4, 5, 6, 7, 8, 9, or more days after the subject contracts the viral infection, and not on the day the subject contracts the viral infection, one day after the subject contracts the viral infection, or two days after the subject contracts the viral infection.

131. 131. The method of claim 130, wherein the effective amount of the gelsolin agent increases the therapeutic effect against a viral infection in a subject compared to the therapeutic effect of a control.

132. 132. The method of claim 131, wherein the control therapeutic effect comprises the therapeutic effect when the gelsolin agent is not administered to the subject.

133. 131. The method of claim 130, wherein the antiviral agent comprises one or more of oseltamivir phosphate, zanamivir, peramivir, and baloxavir marboxil.

134. The method of claim 130, wherein 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% greater than the therapeutic effect of the control.

135. 131. The method of claim 130, wherein the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of a gelsolin molecule.

136. 131. The method of claim 130, wherein the gelsolin molecule is plasma gelsolin (pGSN).

137. 137. The method of claim 135 or 136, wherein the gelsolin molecule is a recombinant gelsolin molecule.

138. The method of claim 131, wherein the therapeutic effect of administering the gelsolin agent reduces the level of viral infection in the subject compared to the level of viral infection in a control, wherein the control level of infection comprises the level of infection when the gelsolin agent is not administered.

139. 139. The method of claim 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 viral infection in the control.

140. The method of claim 139, wherein the level of viral infection in the subject is determined, and the means of determining 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.

141. 141. The method of claim 140, wherein the physiological symptoms include one or more of fever, fatigue, weight loss, and death.

142. The assay detects the presence, absence, and severity of a viral infection in a biological sample from a subject.

141. The method of claim 140, comprising means for detecting the level and / or the amount of the protein.

143. 141. The method of claim 140, wherein administering an effective amount of the gelsolin agent increases the survival chance of the subject compared to the survival chance of a control.

144. 144. The method of claim 143, wherein the control viability is the viability in the absence of administration of the gelsolin agent.

145. 144. The method of claim 143, wherein the increase in subject survival probability 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% greater than the survival probability of the control.

146. 131. The method of claim 130, wherein the means of administration of the gelsolin agent is selected from oral, sublingual, buccal, intranasal, intravenous, inhalation, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intrasynovial, and intraocular administration.

147. 131. The method of claim 130, wherein the subject is a mammal, optionally a human.

148. The method of any one of claims 130 to 147, further comprising treating the subject with an antiviral agent one or more days prior to administering the gelsolin agent to the subject, wherein the antiviral agent is administered on one or more of the following days: the day the subject becomes infected with a viral infection, one day after the subject becomes infected with a viral infection, and two days after the subject becomes infected with a viral infection.

149. The method of claim 148, wherein synergistically effective amounts of a gelsolin agent and an antiviral agent are each administered to a subject and have a synergistic therapeutic effect against a viral infection compared to the therapeutic effect of a control, and the antiviral agent is administered in a clinically acceptable amount.

150. 150. The method of claim 149, wherein the control comprises the therapeutic effect of administering a clinically tolerated amount of an antiviral agent administered without administering a gelsolin agent.

151. 151. The method of claim 150, wherein the clinically tolerated amount of the antiviral agent is an amount less than the maximum tolerated dose (MTD) of the antiviral agent.

152. 152. The method of claim 151, wherein the MTD of the antiviral agent is the highest possible, yet tolerable, dose level of the antiviral agent for the subject.

153. 153. The method of claim 152, wherein the MTD of the antiviral agent is determined based at least in part on a preselected clinically limiting toxicity for the antiviral agent.

154. 150. The method of claim 149, wherein the synergistically effective amounts of the gelsolin agent and the antiviral agent reduce the minimum effective dose (MED) of the antiviral agent in the subject.

155. The method of claim 154, wherein the MED is the lowest dose level of an antiviral agent that provides a clinically significant response in mean efficacy, which response is statistically significantly greater than the response provided by a control that does not include said dose of the antiviral agent.

156. The method of claim 148, wherein the means of administration of the gelsolin agent and the antiviral agent are independently selected from oral, sublingual, buccal, intranasal, inhalation, intravenous, intramuscular, intrathecal, intraperitoneal, subcutaneous, intradermal, topical, rectal, vaginal, intrasynovial, and intraocular administration.

Images