Combination therapy for inhibition of ebolavirus

Abstract

Combination therapies for the inhibition of single-stranded RNA virus replication, such as ebolaviruses are disclosed herein.

Description

[0001] COMBINATION THERAPY FOR INHIBITION OF EBOLAVIRUS

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of and priority to U.S. Provisional Patent Application No. 63 / 612,418, filed December 20, 2023 and U.S. Provisional Patent Application No. 63 / 725,183 filed November 26, 2024, the entirety of these applications is hereby incorporated herein by reference.

[0004] BACKGROUND

[0005] According to the Centers for Disease Control and Prevention, Ebola disease is a rare and often deadly illness, caused by infection by one of a group of four viruses, known as ebolaviruses, that are found primarily in sub-Saharan Africa and are known as: Zaire, Sudan, Tai Forest (formerly Cote d’Ivoire) and Bundibugyo.

[0006] SUMMARY

[0007] Combination therapies for the inhibition of single-stranded RNA virus replication, such as ebolaviruses are disclosed herein. In some embodiments, the combination therapy comprises Opaganib and Remdesivir. In other embodiments, the combination therapy comprises Upamostat and Remdesivir. In other embodiments, the combination therapy comprises Opaganib and Upamostat. Compositions and methods for treating symptomatic and / or asymptomatic infections of ebolaviruses are disclosed herein.

[0008] Aspects of the disclosure relate to a method of treating a human infected with or exposed to an Ebolavirus, the method comprising administering to the human, for a suitable period of time, an effective amount of 3 -(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir. In some embodiments, the effective amount of 3-(4-chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof is administered orally. In some embodiments, the effective amount of 3-(4-chlorophenyl)- adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 9.5 mg / kg to about 15.5 mg / kg daily. In some embodiments, the effective amount of3-(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 10.5 mg / kg to about 14.5 mg / kg daily. In some embodiments, the effective amount of 3-(4-chlorophenyl)-adamantane- 1 - carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 11.5 mg / kg to about 13.5 mg / kg daily. In some embodiments, the effective amount of

[0009] 3-(4-chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 15.0 mg / kg to about 20.0 mg / kg daily. In some embodiments, the method comprises administering orally a solid dosage form comprising the effective amount of the 3-(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof. In some embodiments, remdesivir is administered intravenously. In some embodiments, the human weighs more than 40 kg and the effective amount of remdesivir ranges from 50 mg to 250 mg daily. In some embodiments, the effective amount of remdesivir ranges from 150 to 250 mg daily on day 1, and 50 mg to 150 mg daily on following days. In some embodiments, the method comprises administering intravenously remdesivir over a period of 5 to 10 days. In some embodiments, the human weighs from 3.5 kg to less than 40 kg and the effective amount of remdesivir ranges from 2.5 mg / kg to 5 mg / kg daily. In some embodiments, the effective amount of remdesivir ranges is 5 mg / kg daily on day 1, and 2.5 mg / kg daily on following days. In some embodiments, the method comprises administering intravenously remdesivir over a period of 5 to 10 days. In some embodiments, the method further comprise confirming if the human is infected with an Ebola virus prior to the administering. In some embodiments, the confirming is performed via a test that detects viral antigens or RNA in a sample of blood. In some embodiments, the confirming is performed via a test that detects viral antigens or RNA in a sample of bodily fluids other than blood.

[0010] Other aspects of the disclosure relate to a method of treating a human infected with or exposed to an Ebola virus, the method comprising administering to the human, for a suitable period of time, an effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino-phenylalanine-

[0011] 4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3- hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir. In some embodiments, the pharmaceutically acceptable salt of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino- phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide is a hydrogen sulfate salt. In some embodiments, the effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof ranges from 200 mg to about 400 mg. In some embodiments, the effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof is about 231 mg. In some embodiments, the effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino- phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof ranges from 50 mg to about 200 mg.

[0012] In some embodiments, remdesivir is administered intravenously. In some embodiments, the human weighs more than 40 kg and the effective amount of remdesivir ranges from 50 mg to 250 mg daily. In some embodiments, the effective amount of remdesivir ranges from 150 to 250 mg daily on day 1, and 50 mg to 150 mg daily on following days. In some embodiments, the method comprises administering intravenously remdesivir over a period of 5 to 10 days. In some embodiments, the human weighs from 3.5 kg to less than 40 kg and the effective amount of remdesivir ranges from 2.5 mg / kg to 5 mg / kg daily. In some embodiments, the effective amount of remdesivir ranges is 5 mg / kg daily on day 1, and 2.5 mg / kg daily on following days. In some embodiments, the method comprises administering intravenously remdesivir over a period of 5 to 10 days. In some embodiments, the method further comprises confirming if the human is infected with an Ebola virus prior to the administering. In some embodiments, the confirming is performed via a test that detects viral antigens or RNA in a sample of blood. In some embodiments, the confirming is performed via a test that detects viral antigens or RNA in a sample of bodily fluids other than blood.

[0013] Other aspects of the disclosure relate to a method of treating a human infected with or exposed to an Ebola virus, the method comprising administering to the human, for a suitable period of time, an effective amount of 3 -(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof, and an effective amount of N- α(2,4,6-triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof. In some embodiments, the effective amount of 3-(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin- 4- ylmethyl)amide or a pharmaceutically acceptable salt thereof is administered orally. In some embodiments, the effective amount of 3-(4-chlorophenyl)- adamantane- 1-carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 9.5 mg / kg to about 15.5 mg / kg daily. In some embodiments, the effective amount of 3-(4- chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 10.5 mg / kg to about 14.5 mg / kg daily. In some embodiments, the effective amount of 3-(4-chlorophenyl)-adamantane- 1-carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 11.5 mg / kg to about 13.5 mg / kg daily. In some embodiments, the effective amount of 3-(4- chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 15.0 mg / kg to about 20.0 mg / kg daily. In some embodiments, the method comprises administering orally a solid dosage form comprising the effective amount of the 3-(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutically acceptable salt of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino- phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide is a hydrogen sulfate salt. In some embodiments, the effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxy carbonylpiperazide, free base or a pharmaceutically acceptable salt thereof ranges from 200 mg to about 400 mg. In some embodiments, the effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxy carbonylpiperazide, free base or a pharmaceutically acceptable salt thereof is about 231 mg. In some embodiments, the effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino- phenylalanine-4-ethoxy-carbonylpiperazi de-hydrochloride, N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof ranges from 50 mg to about 200 mg. In some embodiments, the method further comprises confirming if the human is infected with an Ebola virus prior to the administering. In some embodiments, the confirming is performed via a test that detects viral antigens or RNA in a sample of blood. In some embodiments,

[0014] The confirming is performed via a test that detects viral antigens or RNA in a sample of bodily fluids other than blood.

[0015] Aspect of the disclosure relates to an active ingredient combination for treating Ebola virus infection comprising an effective amount of 3 -(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir. In some embodiments, the combination is used to inhibit Ebola virus in a patient.

[0016] Other aspect of the disclosure relates to an active ingredient combination for treating Ebola virus infection comprising: an effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3- amidino-phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir. In some embodiments, the pharmaceutically acceptable salt of N-α-(2,4,6-triisopropylphenylsulfonyl)-3- hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide is a hydrogen sulfate salt. In some embodiments, the combination is used to inhibit Ebola virus in a patient.

[0017] Other aspect of the disclosure relates to an active ingredient combination for treating Ebola virus infection comprising: an effective amount of 3-(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof, and an effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy- carbonylpiperazide-hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino- phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof. In some embodiments, the combination is used to inhibit Ebola virus in a patient.

[0018] BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows plate layout of the Opaganib and Remdesivir combination study set-up according to some embodiments.

[0019] FIG. 2 shows Opaganib and Remdesivir dose response curves and results according to some embodiments.

[0020] FIG. 3 shows in vitro efficacy Opaganib and Remdesivir combination studies (% inhibition, % cell loss and % inhibition not due to cell death) according to some embodiments.

[0021] FIG. 4 is a topology synergy map showing significant synergy peaks were observed for Opaganib and Remdesivir according to some embodiments.

[0022] FIG. 5 is a topology synergy map showing significant synergy peaks were observed for Opaganib and Remdesivir according to some embodiments.

[0023] FIG. 6 shows plate layout of the Upamostat and Remdesivir combination study set-up according to some embodiments.

[0024] FIG. 7 shows potency of individual drugs Upamostat and Remdesivir against EBOV.

[0025] FIG. 8 shows in vitro efficacy Upamostat and Remdesivir combination studies (% inhibition, % cell loss and % inhibition not due to cell death) according to some embodiments.

[0026] FIG. 9 is a topology synergy map showing significant synergy peaks were observed for Upamostat and Remdesivir according to some embodiments.

[0027] FIG. 10 is a topology synergy map showing significant synergy peaks were observed for Upamostat and Remdesivir according to some embodiments.

[0028] FIG. 11 shows dose response curves to determine the potency (EC50), cytotoxicity (CC50) and selectivity index (SI) values.

[0029] FIG. 12 is a topology synergy map showing significant synergy peaks were observed for Opaganib and Remdesivir for % infection inhibition. Topology shown is corrected Inhibition (Ic).

[0030] FIG. 13 shows Opaganib and Upamostat dose curve. Upamostat and Opaganib were tested at starting concentration of 60 μM. Dose response involved three-fold serial dilution, eight different concentrations and four technical replicates either alone or in combination. This combination study was performed in HeLa cells. FIG. 14 is a topology synergy map showing significant synergy peaks were observed for Opaganib and Upamostat for % infection inhibition. Topology shown is corrected Inhibition (Ic).

[0031] FIG. 15 is a topology synergy map showing significant synergy peaks were observed for Opaganib and Upamostat for % corrected infection inhibition. Topology shown is corrected Inhibition (Ic) (not contributed due to cell loss).

[0032] FIG. 16 shows survival curves of mice infected with maEBOV and treated with different doses of Opaganib or drug diluent (vehicle).

[0033] DETAILED DESCRIPTION

[0034] According to the Centers for Disease Control and Prevention (CDC), Ebola disease is a rare and often deadly illness, caused by infection by one of a group of four viruses, known as ebolaviruses, that are found primarily in sub-Saharan Africa and are known as: Zaire, Sudan, Tai Forest (formerly Cote d’Ivoire) and Bundibugyo. Transmission of the disease is mostly through contact with an infected animal (bat or nonhuman primate), or a sick or dead person infected with an ebolavirus. The course of the illness typically progresses from “dry” symptoms initially (such as fever, aches and pains, and fatigue), and then progresses to “wet” symptoms (such as diarrhea, vomiting and unexplained hemorrhaging, bleeding or bruising) as the person becomes sicker. There are currently only two FDA-approved therapies to treat EVD caused by the Ebola virus, species Zaire ebolavirus, in adults and children; Inmazeb™, a combination of three monoclonal antibodies and Ebanga™, a single monoclonal antibody. Both are intravenously infused direct acting monoclonal antibody antivirals that bind to glycoproteins on the Ebola virus’s surface to prevent the virus from entering a person’s cells. There is an urgent need for host-directed small molecule therapies that may be effective against multiple strains of ebolavirus, less likely to be impacted by viral mutation, and that are easy to store, distribute and administer, especially in areas where healthcare services and infrastructures may be sub-optimal.

[0035] As used herein, the term “agent” refers to a compound having a pharmacological activity - an effect of the agent on an individual. The terms “agent,” “compound,” and “drug” are used interchangeably herein . A “patient” or an “individual” refers to any animal, such as a primate. In an embodiment, the primate is a non-human primate. In an embodiment, the primate is a human primate. Any animal can be treated using the methods and composition of the present disclosure.

[0036] As used herein, the term “synergistic effect” refers to the coordinated or correlated action of two or more agents of the present disclosure so that the combined action is greater than the sum of each acting separately. In an embodiment, agents of the present disclosure, when administered together as part of a treatment regimen, provide a therapeutic synergy without accompanying synergistic side effects (e.g., but not limited to, cross-reacting agents).

[0037] As used herein, the term “treat” is meant to administer one or more agents of the present disclosure to measurably inhibit the replication of a virus in vitro or in vivo, to measurably decrease the load of a virus in a cell in vitro or in vivo, or to reduce at least one symptom associated with having a filovirus-mediated disease in a patient. Desirably, the inhibition in replication or the decrease in viral load is at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, as determined using a suitable assay. Assays that monitor replication of viruses include, but are not limited to, cytopathic viral assays, reporter-virus and reporter-cell assays, viral replicon assays, and gene-targeted viral assays. In an embodiment, an assay that measures CD8 T cell-mediated inhibition of filovirus replication is used to measure the slow or stop in the replication of a virus. Viral load testing can be carried out using nucleic acid amplification based tests (NATs or NAATs) and non-nucleic acid-based tests on blood plasma samples to determine the quantity of virus in a given volume including viral RNA levels in plasma and tissue and total viral DNA. Alternatively, in certain embodiments, treatment is observed by a trained physician as an appreciable or substantial relief of symptoms in a patient with a filovirus-mediated disease. Typically, a decrease in viral replication is accomplished by reducing the rate of RNA polymerization, RNA translation, protein processing or modification, or by reducing the activity of a molecule involved in any step of viral replication (e.g., proteins or coded by the genome of the virus or host important for viral replication). In an embodiment, the term “treat” refers to the ability of an agent or agents of the present disclosure to inhibit or suppress replication of a virus, such as an RNA virus. In an embodiment, the term “treat” refers to the ability of an agent or agents of the present disclosure to inhibit the cytopathic effect during a RNA virus infection. By an “effective amount” is meant the amount of an agent or agents of the present disclosure, alone or in combination with another therapeutic regimen, required to treat a patient with a viral disease (e.g., any virus described herein including an Ebola virus or Marburg virus) in a clinically relevant manner. A sufficient effective amount of an agent or agents used to practice the present disclosure for therapeutic treatment of conditions caused by a virus varies depending upon the manner of administration, the age, body weight, and general health of the patient. Ultimately, the prescribers will decide the appropriate amount and dosage regimen. In a combination therapy of the disclosure, the effective amount of an agent may be less than the effective amount if the agent were administered in a non-combinatorial (single-agent) therapy. Additionally, an effective amount may be an amount of an agent in a combination therapy of the disclosure that is safe and efficacious in the treatment of a patient having a viral disease over each agent alone as determined and approved by a regulatory authority (such as the U.S. Food and Drug Administration).

[0038] By "more effective" is meant that a treatment exhibits greater efficacy, or is less toxic, safer, more convenient, or less expensive than another treatment with which it is being compared. Efficacy may be measured by a skilled practitioner using any standard method that is appropriate for a given indication.

[0039] By a “filovirus” is meant a virus belonging to the family Fillovirdae. Exemplary filoviruses are Ebola virus and Marburg virus.

[0040] “Ebola” or “Ebola hemorrhagic fever” is a disease caused by infection with one of the Ebolavirus strains. Ebola can cause disease in humans and nonhuman primates (monkeys, gorillas, and chimpanzees). Ebola disease in humans is caused by four of five viruses in the genus Ebolavirus. The four are Bundibugyo virus (BDBV), Sudan virus (SUDV), Tai Forest virus (TAFV), and one called, simply, Ebola virus (EBOV, formerly Zaire Ebola virus). The fifth virus, Reston virus (RESTV), is not thought to cause disease in humans, but has caused disease in other primates. These five viruses are closely related to marburgviruses. Marburg virus disease (MVD) is a severe illness of humans and non-human primates caused by either of the two marburgviruses, Marburg virus and Ravn virus.

[0041] As used herein, the term “a suitable period of time” refers to the period of time starting when a patient begins treatment for a diagnosis of ssRNA viral infection (e.g., but not limited to, Ebola) using a method of the present disclosure, throughout the treatment, and up until when the patient stops treatment due to either a reduction in symptoms associated with ssRNA viral infection (e.g., but not limited to, Ebola) or due to a laboratory diagnosis indicating that the ssRNA viral infection (e.g., but not limited to, Ebola) is under control. In an embodiment, a suitable period of time is one (1) week. In an embodiment, a suitable period of time is between one (1) week and two (2) weeks. In an embodiment, a suitable period of time is two (2) weeks. In an embodiment, a suitable period of time is between two (2) weeks and three (3) weeks. In an embodiment, a suitable period of time is three (3) weeks. In an embodiment, a suitable period of time is between three (3) weeks and four (4) weeks. In an embodiment, a suitable period of time is four (4) weeks. In an embodiment, a suitable period of time is between four (4) weeks and five (5) weeks. In an embodiment, a suitable period of time is five (5) weeks. In an embodiment, a suitable period of time is between five (5) weeks and six (6) weeks. In an embodiment, a suitable period of time is six (6) weeks. In an embodiment, a suitable period of time is between six (6) weeks and seven (7) weeks. In an embodiment, a suitable period of time is seven (7) weeks. In an embodiment, a suitable period of time is between seven (7) weeks and eight (8) weeks. In an embodiment, a suitable period of time is eight (8) weeks.

[0042] As used herein, the term “cytopathic effects” refers to the changes in cell morphology due to a viral infection.

[0043] As used herein, the terms “cytopathogenesis” or “pathogenesis” includes inhibition of host cell gene expression and includes other cellular changes that contribute to viral pathogenesis in addition to those changes that are visible at the microscopic level.

[0044] The term " in vitro " as used herein refers to procedures performed in an artificial environment, such as for example, without limitation, in a test tube or cell culture system. The skilled artisan will understand that, for example, an isolate SK enzyme may be contacted with a modulator in an in vitro environment. Alternatively, an isolated cell may be contacted with a modulator in an in vitro environment.

[0045] The term “ in vivo" as used herein refers to procedures performed within a living organism such as, without limitation, a human, monkey, mouse, rat, rabbit, bovine, equine, porcine, canine, feline, or primate. Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting 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, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50.

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

[0047] Unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example, within 2 standard deviations of the mean. “About” can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from context, all numerical values provided herein are modified by the term “about”.

[0048] The recitation of an embodiment for a variable or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

[0049] Any compositions or methods provided herein can be combined with one or more of any of the other compositions and methods provided herein.

[0050] Some small viruses carry their genome as single-stranded DNA (ssDNA) molecules. These viruses have a simple genome: one gene for a viral nucleocapsid protein and another gene for a DNA replication enzyme. The virus with a ssDNA genome also faces a serious replication problem in the host cell. When introduced into cells, these genomes cannot be used to make viral proteins because the only template for transcription is double-stranded DNA. For this reason, the first step after infection is the conversion of the viral ssDNA into dsDNA using host cell DNA polymerase. In some of these viruses, the 3' end of the viral DNA folds back and forms dsDNA by base-pairing with an internal sequence. In this way, the primer is built into the genome and the 3' end can be extended to create dsDNA that serves as a template for transcription. The resulting transcripts are translated to make the viral proteins, the replicated viral DNA is converted back into a ssDNA genome, and the virion is packaged for export.

[0051] RNA viruses can be classified according to the sense or polarity of their RNA into negative- sense (-) and positive-sense (+) RNA viruses. The largest family of viruses is the single stranded negative-sense (-) RNA (“ssRNA”) family of viruses. Their viral RNA genome cannot be directly translated, instead the (-) strand is complementary to the viral mRNAs that need to be produced and translated into viral proteins. At the time of this disclosure, one order and eight families are recognized in this group. There are also a number of unassigned species and genera:

[0052] • Order Mononegavirales o Family Bornaviridae — Boma disease virus o Family Filoviridae — includes Ebola virus, Marburg virus o Family Paramyxoviridae — includes Measles virus, Mumps virus, Nipah virus, Hendra virus, RespiratorySyncytial Virus (RSV), and Newcastle Disease Virus (NDV) o Family Rhabdoviridae — includes Rabies virus o Family Nyamiviridae — includes Nyavirus

[0053] • Unassigned families: o Family Arenaviridae — includes Lassa virus o Family Bunyaviridae — includes Hantavirus, Crimean-Congo hemorrhagic fever o Family Ophioviridae o Family Orthomyxoviridae — includes Influenza viruses

[0054] • Unassigned genera: o Genus Deltavirus — includes Hepatitis D virus o Genus Dichorhavirus o Genus Emaravirus o Genus Nyavirus — includes Nyamanini and Midway viruses o Genus Tenuivirus o Genus Varicosavirus

[0055] • Unassigned species: o Taastrup virus o Sclerotinia sclerotiorum negative-stranded RNA virus 1

[0056] Despite decades of efforts by researchers to develop an effective, approved, and available filovirus treatment for individuals, currently there are no United States Food and Drug Administration-approved vaccines or therapeutics for treatment of infection with filovirus diseases.

[0057] Positive-sense viral RNA is similar to mRNA and thus can be immediately translated by the host cell. Negative-sense viral RNA is complementary to mRNA and thus must be converted to positive-sense RNA by an RNA polymerase before translation. As such, purified RNA of a positive-sense virus can directly cause infection though it may be less infectious than the whole virus particle. Purified RNA of a negative- sense virus is not infectious by itself as it needs to be transcribed into positive-sense RNA; each virion can be transcribed to several positive-sense RNAs. Amtisense RNA viruses resemble negative-sense RNA viruses, except they also translate genes from the positive strand. Examples of positive-strand RNA viruses include, but are not limited to, polio virus, Coxsackie virus, and echovirus. Examples of negative- strand RNA viruses include, but are not limited to, influenza virus, measles viruses, and rabies virus.

[0058] The largest family of viruses is the (-) ssRNA family of viruses. Their viral RNA genome cannot be directly translated, instead the (-) strand is complementary to the viral mRNAs that need to be produced and translated into viral proteins. Nature has created hundreds of different (-) ssRNA viruses ranging from the measles and influenza viruses to the rabies and Ebola viruses. Members of this class of virus include Ebola virus and members of the influenza family of viruses.

[0059] Ebola, previously known as Ebola hemorrhagic fever, is a disease caused by infection with one of the Ebola virus strains. Ebola can cause disease in humans and nonhuman primates (monkeys, gorillas, and chimpanzees). Ebola disease in humans is caused by four of five viruses in the genus Ebolavirus. The four are Bundibugyo virus (BDBV), Sudan virus (SUDV), Tai Forest virus (TAFV), and one called, simply, Ebola virus (EBOV, formerly Zaire Ebola virus). The fifth virus, Reston virus (RESTV), is not thought to cause disease in humans, but has caused disease in other primates. These five viruses are closely related to marburgviruses. Currently, no specific therapy is available that has demonstrated efficacy in the treatment of Ebola.

[0060] Ebolaviruses contain single-strand, non-infectious RNA genomes. Ebolavirus genomes are approximately 19 kilobase pairs long and contain seven genes in the order 3'-UTR-NP-VP35- VP40-GP-VP 30-VP24-L-5' -UTR. The genomes of the five different ebolaviruses (BDBV, EBOV, RESTV, SUDV, and TAFV) differ in sequence and the number and location of gene overlaps. In general, ebolavirions are 80 nanometers (nm) in width and may be as long as 14,000 nm. In general, the median particle length of ebolaviruses ranges from 974 to 1,086 nm (in contrast to marburgvirions, whose median particle length was measured at 795-828 nm), but particles as long as 14,000 nm have been detected in tissue culture.

[0061] The viral matrix protein 40 (VP40) is the most abundant protein found in the virions, in infected cells, and also inside the viral nucleocapsid. The nucleoprotein (NP) is associated with the viral genome and assembled into a helical nucleocapside (NC) along with polymerase cofactor (VP35), the transcription activator (VP30), and the RNA-dependent RNA polymerase (L). The viral proteins that comprise the NC catalyze the replication and transcription of the viral genome. A minor viral matrix protein, VP24 is also required for NC assembly. If NP is expressed alone in cells, it assembles together with cellular RNA to form a loose coil-like structure. When NP is co- expressed with VP24 and VP35, NC-like structures are formed in the cytoplasm that are morphologically indistinguishable from those seen in infected cells. It has been shown that VP24 and the viral matrix protein VP40 reduce the transcription and replication efficiencies of the EBOV genome, suggesting that VP24 and VP40 are important for the conversion from a transcription and replication-competent NC to one that is ready for viral assembly. VP40 plays a role in the formation and release of the enveloped, filamentous virus-like particles (VLPs) even when expressed alone. NC-like structures are incorporated into VLPs when VP40 is co-expressed with NP, VP35, and VP24, suggesting that a direct interaction between VP40 and NP is important for the recruitment of NC-like structures to the budding site, the plasma membrane. The interaction between VP40 and NP is also required for the formation of condensed NC-like structures. GP is a surface glycoprotein that forms spikes on virions and plays a crucial role in virus entry into cells by mediating receptor binding and fusion.

[0062] The ebolavirus life cycle begins with virion attachment to specific cell-surface receptors, followed by fusion of the virion envelope with cellular membranes and the concomitant release of the virus nucleocapsid into the cytosol. Ebolavirus' structural glycoprotein (known as GP1,2) is responsible for the virus' ability to bind to and infect targeted cells. The viral RNA polymerase, encoded by the L gene, partially uncoats the nucleocapsid and transcribes the genes into positive- strand mRNAs, which are then translated into structural and nonstructural proteins. The most abundant protein produced is the nucleoprotein, whose concentration in the cell determines when L switches from gene transcription to genome replication. Replication results in full-length, positive-strand antigenomes that are, in turn, transcribed into negative-strand virus progeny genome copy. Newly synthesized structural proteins and genomes self-assemble and accumulate near the inside of the cell membrane. Virions bud off from the cell, gaining their envelopes from the cellular membrane they bud from. The mature progeny particles then infect other cells to repeat the cycle. The Ebola virus genetics are difficult to study due to its virulent nature.

[0063] Ebola is a filamentous, enveloped, negative-sense RNA strand virus in the family of Filoviridae. The RNA-dependent RNA-polymerase of Ebola virus shares significant sequence homology to other negative-strand RNA viruses, required for both viral transcription and replication of the viral genome. However, RNA-dependent RNA-polymerase requires a host factor and viral proteins cooperating to accomplish replication and transcription. One of the reasons why Ebola is so deadly is due to its ability to circumvent the immune system while at the same time pro-actively destroying the human body, as a result the immune system is not able to gather a cohesive effort to fight off the disease. During the infection, monocytes / macrophages in the lymphoid tissues are early and sustained targets of this deadly virus. During the viral infection, large amounts of proinflammatory cytokines such as tumor necrosis factor (TNF-α ) are secreted from infected macrophages and cause disruption of the endothelial barrier. Macrophages and Dendritic cells play a central role in inducing the observed clinical feature of Ebola’s hemorrhagic fever. Secreted cytokines, chemokines, and other mediators alter the blood vessel functions, promote and recruit an influx of inflammatory cells, including additional monocytes / macrophages to the site of the infection. The virus released from the infected macrophages and dendritic cells spread to similar cells throughout the body and to parenchymal cells in many organs, resulting in multifocal tissue necrosis. The ability of the host to develop an effective adaptive immune response is weakened by massive lymphocyte apoptosis, a phenomenon also seen in bacterial sepsis. Ebola infection also induces lymphocyte apoptosis, although the virus does not replicate in lymphocytes. Recent studies indicate that NK (Natural Killer) cells and CD4+ and CD8+ lymphocytes are the principal cell types affected in Ebola-infected macaques monkeys.

[0064] An Ebola subject who shows symptoms (i.e., is symptomatic) including, but not limited to, high fever, headache, joint and muscle aches, sore throat, weakness, stomach pain, lethargy, and lack of appetite, can undergo blood and / or tissue tests to confirm an Ebola diagnosis. An Ebola subject who does not show symptoms (i.e., is asymptomatic) can undergo blood and / or tissue tests to confirm an Ebola diagnosis. These asymptomatic subjects may have markers in their blood indicating they carry the disease, but they are totally asymptomatic.

[0065] A ssRNA virus infected subject who shows symptoms (i.e., is symptomatic) including, but not limited to, high fever, headache, joint and muscle aches, sore throat, weakness, stomach pain, lethargy, and lack of appetite, can undergo blood and / or tissue tests to confirm a diagnosis of a ssRNA virus infection. A ssRNA virus infected subject who does not show symptoms (i.e., is asymptomatic) can undergo blood and / or tissue tests to confirm a diagnosis of ssRNA virus infection. These asymptomatic subjects may have markers in their blood indicating they carry the disease, but they are totally asymptomatic.

[0066] In an embodiment, a subject can be tested for a ssRNA viral infection (e.g., but not limited to, Ebola) within a few days after symptoms begin, or after treatment according to the present disclosure, by collecting a blood or other body fluid sample and testing the sample for detection of viral antigens or RNA in blood and other body fluids using, for example, an antigen-capture enzyme-linked immunosorbent assay (ELISA), using an IgM ELISA (to determine whether the subject has IgM antibodies), using an IgG ELISA (to determine whether the subject has IgG antibodies), using polymerase chain reaction (PCR), or by virus isolation.

[0067] The present disclosure identifies agents and combinations of agents having inhibitory activity against a model filovirus. The present disclosure features compositions and methods for the treatment, of filovirus-mediated disease, e.g., one caused by an Ebola virus or Marburg virus.

[0068] In an embodiment, the present disclosure describes compositions and methods for treating a patient with a filovirus-mediated disease, for example a disease caused by Ebola virus.

[0069] To generate effective therapies against highly pathogenic as well as chronic, persistent RNA viruses, it is important to explore a combination of two or more drugs that act upon different mechanism of action, which will help with efficacy and lowering toxicity of drugs if they are synergistic.

[0070] The method includes administering to the patient a first agent selected from the agents of Table 1, or an analog thereof, in an amount that is effective to treat the patient. In an embodiment, the method further includes administering a second agent selected from the agents of Table 1. In an embodiment, the method further includes administering a third agent selected from the agents of Table 1 Table 1

[0071] When the methods include administering to a patient more than one active agent, the agents may be administered within 7, 6, 5, 4, 3, 2 or 1 days; within 24, 12, 6, 5, 4, 3, 2 or 1 hours, within 60, 50, 40, 30, 20, 10, 5 or 1 minutes; or substantially simultaneously. The methods of the disclosure may include administering one or more agents to the patient by oral, systemic, parenteral, topical, intravenous, inhalational, or intramuscular administration. In an embodiment, the methods of the disclosure include administering one or more agents to the patient by oral administration.

[0072] In an embodiment, the present disclosure describes a composition including two or more agents selected from the agents of Table 1. In an embodiment, the two or more agents are present in amounts that, when administered together to a patient with a filovirus-mediated disease such as a disease caused by Ebola virus, are effective to treat the patient. In an embodiment, the composition consists of active ingredients and excipients, and the active ingredients consist of two or more agents selected from agents of Table 1.

[0073] In an embodiment, the present disclosure describes a composition including three or more agents selected from the agents of Table 1. In an embodiment, the three or more agents are present in amounts that, when administered together to a patient with a filovirus-mediated disease such as a disease caused by Ebola virus, are effective to treat the patient. In an embodiment, the composition consists of active ingredients and excipients, and the active ingredients consist of three or more agents selected from agents of Table 1.

[0074] Active ingredients or agents useful in the invention include those described herein in any of their pharmaceutically acceptable forms, including isomers, salts, solvates, and polymorphs thereof, as well as racemic mixtures and prodrugs. According to aspects illustrated herein, combination therapies of the present disclosure is suitable to inhibit the viral replication machinery of a ssRNA virus, and is also suitable to inhibit the cytopathic effect during a ssRNA virus infection.

[0075] According to aspects illustrated herein, a treatment regimen of the present disclosure is suitable to inhibit the viral replication machinery of a ssRNA virus, and is also suitable to inhibit the cytopathic effect during a ssRNA virus infection.

[0076] Small Hsps interact with a large number of client proteins that are essential to many cellular processes. For example, Hsp90 interacts with over 200 polypeptides in order to modulate their activity and / or half-life. Hsp90, HspBl, and probably other small Hsps, are global regulators of cell systems. Hsp90 is a host factor for the replication of negative strand viruses and is responsible for proteins folding properly, intracellular disposition, stabilizing proteins against heat stress, and also proteolytic turnover of many essential regulators of cell growth and differentiation.

[0077] Upamostat (“WX-671” or “Mesupron”) inhibits the urokinase-type plasminogen activator

[0078] (uPA) system.

[0079] The drug's full chemical description is (S)-ethyl 4-(3-(3-(N- hydroxycarbamimidoyl)phenyl)-2-(2,4,6-triisopropylphenylsulfonamido)propanoyl) piperazine - 1 -carboxylate (or N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxycarbonylpiperazide)). In an embodiment, the compound is N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide present as a salt. In an embodiment, the compound is N-α-(2,4,6-triisopropylphenylsulfonyl)-3- hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide present as a sulfate or hydrogen sulfate salt. In some embodiments, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino- phenylalanine-4-ethoxycarbonylpiperazide is (L)- or (D)-enantiomers, E- or (Z)-isomers or (E / Z)- mixtures, and as free bases or as salts thereof. Upamostat is a serine protease inhibitor. After oral administration, serine protease inhibitor WX-671 is converted to the active Nα-(2,4,6- triisopropylphenylsulfonyl)-3-amidino-(L)-phenylalanine-4-ethoxycarbonylpiperazide (“WX- UK1”), which inhibits several serine proteases, particularly uPA.

[0080] In an embodiment, WX-671 exists as a hydrogen sulfate salt. The serine protease inhibitor Upamostat can potentially inhibit replication of viral RNA.

[0081] An oral formulation of the present disclosure including Upamostat can be used an approach for a therapeutic ssRNA viral infection.

[0082] In an embodiment, the compound is N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino- phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride and in an injectable form to be delivered intravenously or intramuscularly.

[0083] In an embodiment, WX-671 is combined with a pharmaceutically-acceptable carrier material. In an embodiment, the WX-671 and optionally the pharmaceutically-acceptable carrier material, are in a unit dosage form suitable for oral administration. In an embodiment, the dosage form is a solid dosage form. In some embodiments, Upamostat is administered orally at a dose of about 0.5 mg / kg to about 1.1 mg / kg. In an embodiment, Upamostat is administered orally at a daily dose of between about 50 mg to about 200 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 200 mg to about 400 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 50 mg to about 100 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 100 mg to about 150 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 150 mg to about 200 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 200 mg to about 250 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 250 mg to about 300 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 300 mg to about 350 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 350 mg to about 400 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 150 mg to about 550 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 200 mg to about 550 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 250 mg to about 550 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 300 mg to about 550 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 350 mg to about 550 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 400 mg to about 550 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 450 mg to about 550 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 500 mg to about 550 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 200 mg to about 550 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 200 mg to about 500 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 200 mg to about 450 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 200 mg to about 350 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 200 mg to about 300 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 200 mg to about 250 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 500 mg to about 1000 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 750 mg to about 1000 mg. In an embodiment, Upamostat is administered orally at a daily dose of between about 500 mg to about 750 mg.

[0084] An aryladamantane compound of the present invention has been shown to be capable of selectively inhibiting SK2 activity in vitro. Without being bound by theory, it is believed that inhibition of sphingosine kinase (SK) may impair viral protein expression and infectious virus production from cells expressing a cellular protein that acts as a receptor for Ebola virus and Marburg virus. According to aspects illustrated herein, there is disclosed a composition comprising an aryladamantane compound, wherein the aryladamantane compound is present in an amount that, when administered to a patient with a filovirus-mediated disease, are effective to treat the patient. In an embodiment, the filovirus is Ebola virus or Marburg virus. According to aspects illustrated herein, a method for treating a patient having filovirus-mediated disease includes administering to the patient a composition comprising an aryladamantane compound in an amount effective to treat the patient. In an embodiment, the filovirus is Ebola virus or Marburg virus.

[0085] The symbol in general represents a bond between two atoms in the chain. Thus CH3O — CH2— CH( R1) — CH3represents a 2-substituted-1-niethoxypropane compound. In addition, the symbol represents the point of attachment of the substituent to a compound. Thus for example aryl(C1-C6)alkyl- indicates an alkylaryl group, such as benzyl, attached to the compound at the alkyl moiety.

[0086] Where multiple substituents are indicated as being attached to a structure, it is to be understood that the substituents can be the same or different. Thus for example “Rmoptionally substituted with 1, 2 or 3 Rqgroups” indicates that Rmis substituted with 1, 2, or 3 Rqgroups where the Rqgroups can be the same or different

[0087] The phrase “optionally substituted” is used interchangeably with the phrase “substituted or unsubstituted”. Unless otherwise indicated, an optionally substituted group may have a substituent at each substitutable position of the group, and each substituent is independent of the other.

[0088] As used herein, the terms “halogen” or “halo” indicate fluorine, chlorine, bromine, or iodine

[0089] The term “heteroatom” means nitrogen, oxygen or sulfur and includes any oxidized form of nitrogen and sulfur, and the quaternized form of any basic nitrogen. Also the term “nitrogen” includes a substitutable nitrogen in a heterocyclic ring As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from nitrogen, oxygen or sulfur, the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR+(as in N-substituted pyrrolidinyl).

[0090] The term “alkyl”, as used herein alone or as part of a larger moiety, refers to a saturated aliphatic hydrocarbon including straight chain, branched chain or cyclic (also called “cycloalkyl”) groups. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, iso-, sec- and iert-butyl, pentyl, hexyl, heptyl, 3-ethylbutyI, and the like. Preferably, the alkyl group has 1 to 20 carbon atoms (whenever a numerical range, e.g. “1-20”, is stated herein, it means that the group, in this case the alkyl group, may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc. up to and including 20 carbon atoms). More preferably, it is a medium size alkyl having 1 to 10 carbon atoms. Most preferably, it is a lower alkyl having 1 to 4 carbon atoms. The cycloalkyl can be monocyclic, or a polycyclic fused system. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cycolpentyl, cyclohexyl, cycloheptyl, cyclooctyl, and adamantyl. The alkyl or cycloalkyl group may be unsubstituted or substituted with 1 , 2, 3 or more substituents. Examples of such substituents including, without limitation, halo, hydroxy, amino, alkoxy, alkylamino, dialkylamino, cycloalkly, aryl, aryloxy, arylalkyloxy, heterocyclic radical, and (heterocyclic radical)oxy. Examples include fluoromethyl, hydroxyethyl, 2,3-dihydroxyethyl, (2- or 3- furanyl)m ethyl, cyclopropyl methyl, benzyloxyethyl, (3-pyridinyl)rnethyl, (2-thienyl)ethyl, hyroxypropyl, aminocyclohexyl, 2-dimethylaminobutyl, methoxymethyl, N-pyridinylethyl, and diethylaminoethyl .

[0091] The term “cycloalkylalkyl”, as used herein alone or as part of a larger moiety, refers to a C3-C10cycloalkyl group attached to the parent molecular moiety through an alkyl group, as defined above. Examples of cycloalkyl alkyl groups include cyclopropylmethyl and cyclopentylethyb

[0092] The term “alkenyl”, as used herein alone or as part of a larger moiety, refers to an aliphatic hydrocarbon having at least one carbon-carbon double bond, including straight chain, branched chain or cyclic groups having at least one carbon-carbon double bond. Preferably, the alkenyl group has 2 to 20 carbon atoms. More preferably, it is a medium size alkenyl having 2 to 10 carbon atoms. Most preferably, it is a lower alkenyl having 2 to 6 carbon atoms. The alkenyl group may be unsubstituted or substituted with 1, 2, 3 or more substituents. Examples of such substituents including, without limitation halo, hydroxy, amino, alkoxy, alkylamino, dialkylamino, cycloalkly, aryl, aryloxy, arylalkyloxy, heterocyclic radical, and (heterocyclic radical)oxy. Depending on the placement of the double bond and substituents, if any, the geometry of the double bond may be entgegen (E) or zusammen (Z), cis, or trans. Examples of alkenyl groups include ethenyl, propenyl, cis-2-butenyl, trans-2-butenyl, and 2-hyroxy-2-propenyl,

[0093] The term “alkynyl”, as used herein alone or as part of a larger moiety, refers to an aliphatic hydrocarbon having at least one carbon-carbon triple bond, including straight chain, branched chain or cyclic groups having at least one carbon-carbon triple bond. Preferably, the alkynyl group has 2 to 20 carbon atoms. More preferably, it is a medium size alkynyl having 2 to 10 carbon atoms. Most preferably, it is a lower alkynyl having 2 to 6 carbon atoms. The alkynyl group may be unsubstituted or substituted with 1, 2, 3 or more substituents. Examples of such substituents including, without limitation, halo, hydroxy, amino, alkoxy, alkylamino, dialkylamino, cycloalkly, aryl, aryloxy, arylalkyloxy, heterocyclic radical, and (heterocyclic radical)oxy. Examples of alkynyl groups include ethynyl, propynyl, 2-butynyl, and 2-hyroxy-3-butylnyl.

[0094] The term “alkoxy”, as used herein alone or as part of a larger moiety, represents an alky! group of indicated number of carbon atoms attached to the parent molecular moiety through an oxygen bridge. Examples of alkoxy groups include, for example, methoxy, ethoxy, propoxy and isopropoxy. Alkoxy radicals may be further substituted with one or more halo atoms, such as fluoro, chloro or bromo, to provide “haloalkoxy” radicals. Examples of such radicals include fluoromethoxy, chloromethoxy, trifluoromethoxy, and fluoroethoxy.

[0095] The term “aryl”, as used herein alone or as part of a larger moiety, refers to an aromatic hydrocarbon ring system containing at least one aromatic ring The aromatic ring may optionally be fused or otherwise attached to other aromatic hydrocarbon rings or non-aromatic hydrocarbon rings. Additionally, the aryl group may be substituted or unsubstituted by various groups such as hydrogen, halo, hydroxy, alkyl, haloalkyl, alkoxy, nitro, cyano, alkylamine, carboxy or alkoxycarbonyl. Examples of aryl groups include, for example, phenyl, naphthyl, 1, 2,3,4- tetrahydronaphthalene, benzodi oxole, and biphenyl. Preferred examples of unsubstituted aryl groups include phenyl and biphenyl Preferred aryl group substituents include hydrogen, halo, alkyl, haloalkyl, hydroxy and alkoxy. The term “heteroalkyl”, as used herein alone or as part of a larger moiety, refers to an alkyl radical as defined herein with one or more heteroatoms replacing a carbon atom with the moiety. Such heteroalkyl groups are alternately referred to using the terms ether, thioether, amine, and the like.

[0096] The term “heterocyclyl”, as used herein alone or as part of a larger moiety, refers to saturated, partially unsaturated and unsaturated heteroatom-containing ring shaped radicals, where the heteroatoms may be selected from nitrogen, sulfur and oxygen. Said heterocyclyl groups may be unsubstituted or substituted at one or more atoms within the ring system. The heterocyclic ring may contain one or more oxo groups.

[0097] The term “heterocycloalkyl”, as used herein alone or as part of a larger moiety, refers to a non-aromatic ring system containing at least one heteroatom selected from nitrogen, oxygen, and sulfur. The heterocycloalkyl ring may be optionally fused to or otherwise attached to other heterocycloalkyl rings and / or non-aromatic hydrocarbon rings. Preferred heterocycloalkyl groups have from 3 to 7 members. Examples of heterocycloalkyl groups include, for example, piperazine, morpholine, piperidine, tetrahydrofuran, pyrrolidine, and pyrazole. Preferred monocyclic heterocycloalkyl groups include piperidyl, piperazinyl, morpholinyl, pyrrolidinyl, thioraorpholinyl, thiazolidinyl, 1,3-dioxolanyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like. Heterocycloalkyl radicals may also be partially unsaturated. Examples of such groups include dihydrothienyl, dihydropyranyl, di hydrofuryl, and dihydrothiazolyl.

[0098] The term “heteroaryl”, as used herein alone or as part of a larger moiety, refers to an aromatic ring system containing at least one heteroatom selected from nitrogen, oxygen, and sulfur The heteroaryl ring may be fused or otherwise attached to one or more heteroaryl rings, aromatic or non-aromatic hydrocarbon rings or heterocycloalkyl rings. Additionally, the heteroaryl group may be unsubstituted or substituted at one or more atoms of the ring system, or may contain one or more oxo groups. Examples of heteroaryl groups include, for example, pyridine, furan, thiophene, carbazole and pyrimidine. Preferred examples of heteroaryl groups include thienyl, benzothienyl, pyridyl, quinolyl, pyrazinyl, pyrimidyl, imidazolyl, benzimidazolyl, furanyl, benzofuranyl, thiazolyl, benzothiazolyl, isoxazolyl, oxadiazolyl, isothiazolyl, benzisothiazolyl, triazolyl, tetrazolyl, pyrrolyl, indolyl, pyrazolyl, benzopyrazolyl, purinyl, benzooxazolyl, and carbazolyl.

[0099] The term “acyl” means an H — C(O) — or alkyl-C(O) — group in which the alkyl group, straight chain, branched or cyclic, is as previously described. Exemplary acyl groups include formyl, acetyl, propanoyl, 2-methylpropanoyl, butanoyl, and caproyl.

[0100] The term “aroyl” means an aryl-C(O) — group in which the aryl group is as previously described. Exemplary aroyl groups include benzoyl and 1- and 2-naphthoyl.

[0101] The term “solvate” means a physical association of a compound of this invention with one or more solvent molecules. This physical association involves varying degress of ionic and covalent bonding, including hydrogen bonding. In certain instances, the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. “Solvate” encompasses both solution-phase and isolatable solvates Exemplary solvates include ethanolates, methanolates, and the like. “Hydrate” is a solvate wherein the solvent molecule(s) is / are H2O.

[0102] Examples of aryladamantane compound of the present invention are generally represented by Formula I, shown below: and pharmaceutically acceptable salts thereof, wherein

[0103] L is a bond or is — C(R3,R4) — ;

[0104] X is— C(R3,R4)N(R5)— , — C(O)N(R4)— , — N(R4)C(O)— , — C(R4,R5)— , — N(R4)— , — O— , — S— — C(O)— , — S(O)2— , — S(O)2N(R4)— or— N(R4)S(O)2— ;

[0105] R1is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, — COOH, —OH, — SH, — S-alkyl, — CN, — NO2, — NH2, — CO2(alkyl), — OC(O)alkyl, carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, or mono or dialkylthiocarbamoyl; R2is H, alkyl, cycloalkyl, cycloalkyl alkyl, alkenyl, alkynyl, heteroalkyl, and, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, — COOH, — OH, — SH, — S-alkyl, — CN, — NO2, — NH2, — CO2(alkyl), — OC(O)alkyl, carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, mono or dialkylthiocarbamoyl, alkyl-S-alky 1, -heteroaryl-aryl, -alkyl-heteroaryl-aryl, — C(O) — NH-aryl, -alkenyl-heteroaryl, — C(O)-heteroaryl, or -alkenyl- heteroaryl-aryl; R3is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, oxo (=O), — COOH, — OH, — SH, — S-alkyl, — CN, — NO2, — NH2, — CO2(alkyl), — OC(O)alkyl, carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, or mono or dialkylthiocarbamoyl; wherein the alkyl and ring portion of each of the above R1, R2, and R? groups is optionally substituted with up to 5 groups that are independently (C1-C6) alkyl, halogen, haloalkyl, — OC(O)(C1-C6alkyl), — C(O)O(C1-C6alkyl), — CONR'R", — OC(O)NR'R", — NR'C(O)R", — CF3, — OCF3, —OH, C1-C6alkoxy, hydroxyalkyl, — CN, — CO2H, — SH, —S-alkyl, — SOR'R", — SO2R', — NO2, or NR'R", wherein R' and R" are independently H or (C1-C6) alkyl, and wherein each alkyl portion of a substituent is optionally further substituted with 1, 2, or 3 groups independently selected from halogen, CN, OH, and NHz; and R4and R5are independently H or alkyl, provided that when R3 and R4are on the same carbon and R3 is oxo, then R4is absent.

[0106] Aryladamantane compounds of Formula I include compounds of formula 1-1 : and pharmaceutically acceptable salts thereof, wherein:

[0107] R1is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl -heterocycloalkyl, acyl, aroyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, — COOH, — OH, — SH, — S-alkyl, — CN, — NO2, — NH2, — CO2(alkyl), — OC(O)alkyl, carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, or mono or dialkylthiocarbamoyl, and

[0108] R2is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, — COOH, — OH, — SH, — S-alkyl, — CN, — NO2, — NH2, — CO2(alkyl), — OC(O)alkyl, carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, mono or dialkylthiocarbamoyl, alkyl-S-alkyl, -heteroaryl-aryl, -alkyl-heteroaryl-aryl, — NH-aryl, -alkenyl-heteroaryl, -heteroaryl, — NH-alkyl, — NH- cycloalkyl, or -alkenyl-heteroaryl-aryl, wherein the alkyl and ring portion of each of the above R1, and R2groups is optionally substituted with up to 5 groups that are independently (C1-C6) alkyl, halogen, haloalkyl, — OC(O)(C1-C6alkyl), — C(O)O(C1-C6alkyl), — CONR'R", — OC(O)NR'R", — NR'C(O)R", — CF3, — OCF3, — OH, C1-C6alkoxy, hydroxyalkyl, — CN, — CO2H, — SH, -S-alkyl, — SOR'R", — SO2R', — NO2, or NR'R", wherein R' and R" are independently H or (C1-C6) alkyl, and wherein each alkyl portion of a substituent is optionally further substituted with 1, 2, or 3 groups independently selected from halogen, CN, OH, NH2.

[0109] Aryladamantane compounds of Formula I include those of formula II: and pharmaceutically acceptable salts thereof, wherein:

[0110] Y is — C(R4,R5)— , — N(R4)— , — O— , or — C(O)— ,

[0111] R1is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, — COOH, — OH, — SH, — S-alkyl, — CN, — NO2, — NH2, — CO2(alkyl), — OC(O)alkyl, carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, or mono or dialkylthiocarbamoyl;

[0112] R2is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, — COOH, — OH, — SH, — S-alkyl, — CN, —NO2, — NH2, — CO2(alkyl), — OC(O)alkyl, carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, mono or dialkylthiocarbamoyl, alkyl-S-alky 1, -heteroaryl-aryl, -alkyl-heteroaryl-aryl, — C(O) — NH-aryl, -alkenyl-heteroaryl, — C(O)-heteroaryl, or -alkenyl- heteroaryl-aryl; R3is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, oxo (=O), — COOH, — OH, — SH, — S-alkyl, — CN, — NO2, — NHz, — CO2(alkyl), — OC(O)alkyl, carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, or mono or dialkylthiocarbamoyl; wherein the alkyl and ring portion of each of the above R1, R2, and R3groups is optionally substituted with up to 5 groups that are independently (C1-C6) alkyl, halogen, haloalkyl, — OC(O)(C1-C6alkyl), — C(O)O(C1-C6alkyl), — CONRR", — OC(O)NRR", — NR'C(O)R", — CF3, — OCF3, —OH, C1-C6alkoxy, hydroxyalkyl, — CN, — CO2H, — SH, —S-alkyl, — SORR”, — SO2R', — NO2, or NR'R", wherein R' and R" are independently H or (C1-C6) alkyl, and wherein each alkyl portion of a substituent is optionally further substituted with 1, 2, or 3 groups independently selected from halogen, CN, OH, NH2; and R4and R5are independently H or alkyl

[0113] Compounds of the formula II include those wherein:

[0114] Y is — C(R4,R5) — or — N(R4) — ,

[0115] R1is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxy alkyI, alkanoyl, — COOH, —OH, ----- SH, — S-alkyl, — CN, — NO2, — NH2, — COi(alkyl), — OC(O)alkyl, carbamoyl, mono or dialkylaminocarbarnoyl, mono or di alkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, or mono or dialkylthiocarbamoyl;

[0116] R2is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyd, alkyl-heterocycloalkyl, acyl, aroyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, — COOH, — OH, — SH, --S-alkyl, ----CN, --NO2, --NH2, — CO2(alkyl), - - OC(O)alkyl, carbamoyl, mono or dialkylaminocarbarnoyl, mono or dialkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, mono or dialkyIthiocarbamoyl, alkyl-S-alkyl, -heteroaryl-aryl, -alkyl-heteroaryl-aryl, — C(O) — NH-aryl, -alkenyl-heteroaryl, — C(O)-heteroaryl, or -alkenyl- heteroaryl -aryl; wherein the alkyl and ring portion of each of the above R1and R2groups is optionally substituted with up to 5 groups that are independently (C1-C6) alkyl, halogen, haloalkyl, — OC(O)(C1-C6alkyl), --C(O)O(C1-C6alkyl), --CONR4R5, ------OC(O)NR4R5, -- NR4C(O)R5, --CF3, --OCF3, -- OH, C1-C6alkoxy, hydroxy alkyl, — CN, — CO2H, — SH, -S-alkyl, — SOR4R5, — SO2R4R5, — NO2, or NR4R5, and wherein each alkyl portion of a substituent is optionally further substituted with 1, 2, or 3 groups independently selected from halogen, CN, OH, NH2;

[0117] R3is H, alkyl, or oxo (=0), and

[0118] R4and R5are independently H or (C1-C6)alkyl.

[0119] Representative formula II compounds include:

[0120] Cmpd Chemical name Y R3 R1 R2

[0121]

[0122] A particularly preferred aryladamantane compound of the present invention is illustrated below and referred to as ABC294640 [3-(4-chlorophenyl)-adantaniane-1-carboxylic acid (pyridin- 4-ylmethyl)amide] (also referred to as Opaganib): In an embodiment, an aryladamantane compound of the present invention is selected from a compound of Formula below: and pharmaceutically acceptable salts thereof, wherein o R1is H, Cl or F, o R2is H or alkyl; o m is 0, 1 or 2; o n is 1, 2, 3, 4 or 5; o each R3is independently H, (O)alkyl --C(O)CH2CH2C(O)OH, R4, ------ C(O)NR5R6, — P(O)(OR7)2or glucosyl, provided that at least one R.i is not H, o wherein

[0123] ■ R4is a natural or unnatural amino acid linked through the carboxyl moiety as an ester,

[0124] ■R5is H or alkyl,

[0125] ■ R6is H or alkyl, and each R7is independently H or alkyl.

[0126] In certain embodiments of the compounds of formula (I) as described above, the moiety is a catechol with substitution at least one catechol — OH. For example, in one embodiment, the

[0127]

[0128] In one particularly preferred embodiment of the compounds of formula (I) as described above, the

[0129] In one especially preferred embodiment of the invention, compounds of formula (I) have R1=Cl,

[0130] R2=H, m=2, n=2, and each R1= — C(O)alkyl, especially — C(O)CH3.

[0131] For example, compounds of the invention include:

[0132] » Acetic acid 2-acetoxy-5-(2-{[3-(4-chlorophenyl)-adamantane-1-carbonyl]- amino]ethyl)phenyl ester;

[0133] « Propionic acid 2-propionyloxy-5-(2-{ [3-(4-chlorophenyl)-adamantane- 1 -carbonyl]- amiiio}ethyl)phenyl ester;

[0134] » Butyric acid 2-butyryloxy-5-(2-{[3-(4-chlorophenyl)-adamantane-1-carbonyl]- amino}ethyl)phenyl ester;

[0135] « Isobutyric acid 5-(2-{[3-(4-chlorophenyl)adamantane-1-carbonyi]amino}ethyl)-2- hydroxyphenyl ester; and

[0136] 2-Amino-3 -methyl -butyric acid 5-(2-{[3-(4-chlorophenyl)adamantane-1- carbonyl]amino} ethyl)-2-hydroxyphenyl ester. A particularly preferred aryladamantane compound of the present invention is illustrated below and referred to as ABC294735 [3 -(4-chlorophenyl)adamantane-1-carboxylic acid [2-(3,4- dihydroxyphenyl)ethyl]amide] :

[0137] In an embodiment, an effective amount of the compound of formula I is between about 15,0 mg / kg / day to about 20 mg / kg / day. In an embodiment, the compound of formula I is administered at a dailv dosage ranging from about 2,5 mg / kg to about 22,5 mg / kg. In an embodiment, the compound of formula I is administered at a daily dosage ranging from about 3.5 mg / kg to about 21.5 mg / kg. In an embodiment, the compound of formula I is administered at a daily dosage ranging from about 4.5 mg / kg to about 20.5 mg / kg. In an embodiment, the compound of formula I is administered at a daily dosage ranging from about 5.5 mg / kg to about

[0138] 19.5 mg / kg. In an embodiment, the compound of formula I is administered at a daily dosage ranging from about 6.5 mg / kg to about 18.5 mg / kg. In an embodiment, the compound of formula I is administered at a daily dosage ranging from about 7.5 mg / kg to about 17.5 mg / kg. In an embodiment, the compound of formula I is administered at a daily dosage ranging from about 8.5 mg / kg to about 16.5 mg / kg. In an embodiment, the compound of formula I is administered at a daily dosage ranging from about 9,5 mg / kg to about 15.5 mg / kg. In an embodiment, the compound of formula I is administered at a daily dosage ranging from about 10.5 mg / kg to about 14.5 mg / kg. In an embodiment, the compound of formula I is administered at a daily dosage ranging from about

[0139] 11.5 mg / kg to about 13.5 mg / kg.

[0140] In an embodiment, the determining step includes measuring, at least two different times during the suitable time period, the viral load using a nucleic acid amplification based test. In an embodiment, the inhibition in viral replication or the decrease in viral load is at least 10% as determined using a nucleic acid amplification based test. In an embodiment, the individual is a human. In an embodiment, the filovirus is Ebola virus or Marburg virus. In an embodiment, the filovirus is Ebola virus. In an embodiment, the compound of formula I is present as a solid dosage form. In an embodiment, the solid dosage form is a capsule.

[0141] In an embodiment, the determining step includes measuring, at least two different times during the suitable time period, the viral load using a nucleic acid amplification based test. In an embodiment, the inhibition in viral replication or the decrease in viral load is at least 10% as determined using a nucleic acid amplification based test. In an embodiment, the individual is a human. In an embodiment, the filovirus is Ebola virus or Marburg virus.

[0142] Solid forms for oral administration may contain pharmaceutically acceptable binders, sweeteners, disintegrating agents, diluents, flavorings, coating agents, preservatives, lubricants, and / or time delay agents. Suitable binders include gum acacia, gelatin, corn starch, gum tragacanth, sodium alginate, carboxymethylcellulose or polyethylene glycol (PEG). Suitable sweeteners include sucrose, lactose, glucose, aspartame or saccharine. Suitable disintegrating agents include corn starch, methylcellulose, polyvinylpyrrolidone, xanthan gum, bentonite, alginic acid or agar. Suitable diluents include lactose, sorbitol, mannitol, dextrose, kaolin, cellulose, calcium carbonate, calcium silicate or dicalcium phosphate. Suitable flavoring agents include peppermint oil, oil of wintergreen, cherry, orange, or raspberry flavoring. Suitable coating agents include polymers or copolymers of acrylic acid and / or methacrylic acid and / or their esters, waxes, fatty alcohols, zein, shellac or gluten. Suitable preservatives include sodium benzoate, vitamin E, alpha-tocopherol, ascorbic acid, methyl paraben, propyl paraben or sodium bisulphite. Suitable lubricants include magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc. Suitable time delay agents include glyceryl monostearate or glyceryl distearate

[0143] Remdesivir

[0144] Remdesivir (2-ethylbutyl (2S)-2-[[[(2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin- 7-yl)-5-cyano-3,4-dihydroxyoxolan-2-yl]methoxy-phenoxyphosphoryl]amino]propanoate) is a carboxylic ester resulting from the formal condensation of the carboxy group of N-[(S)- {[(2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano-3,4- dihydroxytetrahydrofuran-2-yl]methoxy}(phenoxy)phosphoryl]-L-alanine with the hydroxy group of 2-ethylbutan-1-oL Remdesivir is a broad-spectrum antiviral prodrug with potent in vitro antiviral activity against a diverse panel of RNA viruses such as Ebola virus, Marburg virus, MERS-CoV and SARS-CoV. It has demonstrated in vitro activity against the Arenaviridae, Flaviviridae, Filoviridae, Paramyxoviridae, Pneumoviridae, and Coronaviridae viral families. It is a carboxylic ester, a pyrrolotriazine, a nitrile, a phosphoramidate ester, a C-nucleoside and an aromatic amine.

[0145] In some embodiments, Remdesivir s administered intravenously. In some embodiments, Remdesivir is administered once daily. In some embodiments, the human weighs at least 40 kg, and the Remdesivir, is administered in the first dose of 150-250 mg on day 1, and administered in a second dose of 50-150 mg on each of the following 4 days. In some embodiments, the second dose of 50-150 mg is administered for an additional 1 to 5 days. In some embodiments, Remdisivir is administered in the first dose of 150-250 mg on day 1, and administered in a second dose of 50- 150 mg on each of the following 4, 5, 6, 7, 8, or 9 days. In some embodiments, Remdisivir is administered in the first dose of 150-250 mg on day 1, and administered in a second dose of 50- 150 mg on each of the following 9 days. In some embodiments, Remdisivir is administered once daily. In some embodiments, Remdisivir is administered intravenously in the first dose of 200 mg on day 1, and administered intravenously in a second dose of 100 mg on each of the following 4 days. In some embodiments, the second dose of 100 mg is administered for an additional 1 to 5 days. In some embodiments, Remdisivir is administered intravenously in the first dose of 200 mg on day 1, and administered intravenously in a second dose of 100 mg on each of the following 4, 5, 6, 7, 8, or 9 days. In some embodiments, Remdisivir is administered intravenously in the first dose of 200 mg on day 1, and administered intravenously in a second dose of 100 mg on each of the following 9 days. In some embodiments, Remdisivir is administered intravenously once daily. In some embodiments, Remdisivir is administered intravenously over from about 30 to about 120 minutes, is administered in the first dose of 150-250 mg on day 1, and administered in a second dose of 50-150 mg on each of the following 4 days. In some embodiments, the second dose of 50- 150 mg is administered for an additional 1 to 5 days. In some embodiments, Remdisivir is administered in the first dose of 150-250 mg on day 1, and administered in a second dose of 50- 150 mg on each of the following 4, 5, 6, 7, 8, or 9 days. In some embodiments, Remdisivir is administered in the first dose of 150-250 mg on day 1, and administered in a second dose of 50- 150 mg on each of the following 9 day. In some embodiments, Remdisivir is administered intravenously over a period ranging from about 30 to about 120 minutes.

[0146] Combination Therapy In some embodiments, administration of ABC294640 has a synergistic activity with remdesivir to inhibit the replication machinery of a ssRNA virus infection. In some embodiments, administration of the combination of ABC294640 and remdesivir hinders, restrains or prevents viral infection.

[0147] In some embodiments, administration of Upamostat has a synergistic activity with remdesivir to inhibit the replication machinery of a ssRNA virus infection. In some embodiments, administration of the combination of Upamostat and remdesivir hinders, restrains or prevents viral infection.

[0148] In some embodiments, administration of ABC294640 and Upamostat has a synergistic activity with remdesivir to inhibit the replication machinery of a ssRNA virus infection. In some embodiments, administration of the combination of ABC294640, Upamostat and remdesivir hinders, restrains or prevents viral infection.

[0149] In some embodiments, administration of ABC294640 has a synergistic activity with an anti-viral drug to inhibit the replication machinery of a ssRNA virus infection. In some embodiments, administration of the combination of ABC294640 and an anti-viral drug hinders, restrains or prevents viral infection.

[0150] In some embodiments, administration of Upamostat has a synergistic activity with an anti- viral drug to inhibit the replication machinery of a ssRNA virus infection. In some embodiments, administration of the combination of Upamostat and an anti-viral drug hinders, restrains or prevents viral infection.

[0151] In some embodiments, administration of ABC294640 and Upamostat has a synergistic activity with an anti-viral drug to inhibit the replication machinery of a ssRNA virus infection. In some embodiments, administration of the combination of ABC294640, Upamostat and an anti- viral drug hinders, restrains or prevents viral infection.

[0152] In some embodiments, administration of ABC294640 has a synergistic activity with Upamostat to inhibit the replication machinery of a ssRNA virus infection.

[0153] In some embodiments, Upamostat is administered orally as a solid dosage form. In some embodiments, Upamostat is administered orally one or more times per day.

[0154] In some embodiments, ABC294640 is administered orally as a solid dosage form. In some embodiments, ABC294640 is administered orally one or more times per day. In some embodiments, anti-viral drugs are administered orally or intravenously. In some embodiments, anti-viral drugs are administered orally or intravenously one or more times per day. In some embodiments, anti-viral drugs are administered intravenously. In some embodiments, the anti-viral drug is remdesivir.

[0155] In an embodiment, the present disclosure relates to methods for treating a subject having a ssRNA viral infection, such as Ebola virus or Marburg virus, by concomitantly administering i) a therapeutically effective amount of an anti-viral drug; and ii) a therapeutically effective amount of ABC294640, Upamostat or a combination thereof.

[0156] In an embodiment, the present disclosure relates to methods for treating a subject having a ssRNA viral infection, such as Ebola virus, by administering i) a therapeutically effective amount of an anti-viral drug; and ii) a therapeutically effective amount of ABC294640. In some embodiments, the i) therapeutically effective amount of an anti-viral drug; and ii) therapeutically effective amount of ABC294640 are administered concomitantly. In other embodiments, the i) therapeutically effective amount of an anti-viral drug; and ii) therapeutically effective amount of ABC294640 are administered sequentially.

[0157] In an embodiment, the present disclosure relates to methods for treating a subject having a ssRNA viral infection, such as Ebola virus, by administering i) a therapeutically effective amount of an anti-viral drug; and ii) a therapeutically effective amount of upamostat. In some embodiments, the i) therapeutically effective amount of an anti-viral drug; and ii) therapeutically effective amount of Upamostat are administered concomitantly. In other embodiments, the i) therapeutically effective amount of an anti-viral drug; and ii) therapeutically effective amount of Upamostat are administered sequentially.

[0158] In an embodiment, the present disclosure relates to methods for treating a subject having a ssRNA viral infection, such as Ebola virus, administering i) a therapeutically effective amount of an anti-viral drug; ii) a therapeutically effective amount of ABC294640; iii) a therapeutically effective amount of Upamostat. In some embodiments, the i) therapeutically effective amount of an anti-viral drug; ii) therapeutically effective amount of ABC294640; and iii) therapeutically effective amount of Upamostat are administered concomitantly. In other embodiments, the i) therapeutically effective amount of an anti-viral drug; ii) therapeutically effective amount of ABC294640; and iii) therapeutically effective amount of Upamostat are administered sequentially. In an embodiment, one, two or more antiviral drugs are administered to the patient in need thereof. In an embodiment, the anti-viral drug is remdesivir. In an embodiment, the anti-viral drug is a direct antiviral agent. In an embodiment, the anti-viral drug is host directed small molecule. In some embodiments, the one, two or more antiviral drugs are administered intravenously. In some embodiments, the one, two or more antiviral drugs are administered by infusion.

[0159] In an embodiment, the present disclosure relates to methods for treating a subject having a ssRNA viral infection, e.g., but not limited to, Ebola virus, by administering a therapeutically effective amount of an aryladamantane compound. In an embodiment the aryladamantane compound is selected from a compound of formula I.

[0160] Aspects of the disclosure relate to a method of treating a human infected with or exposed to an Ebola virus, the method comprising administering to the human, for a suitable period of time, an effective amount of 3 -(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyljamide or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir. In some embodiments, the effective amount of 3-(4-chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4- ylmethyljamide or a pharmaceutically acceptable salt thereof is administered orally. In some embodiments, the effective amount of 3-(4-chlorophenyl)- adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 9.5 mg / kg to about 15.5 mg / kg daily. In some embodiments, the effective amount of 3-(4-chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 10.5 mg / kg to about 14.5 mg / kg daily. In some embodiments, the effective amount of 3-(4-chlorophenyl)-adamantane- 1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 11.5 mg / kg to about 13.5 mg / kg daily. In some embodiments, the effective amount of 3-(4-chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 15.0 mg / kg to about 20.0 mg / kg daily. In some embodiments, the method comprises administering orally a solid dosage form comprising the effective amount of the 3-(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyljamide or a pharmaceutically acceptable salt thereof. In some embodiments, remdesivir is administered intravenously. In some embodiments, the human weighs more than 40 kg and the effective amount of remdesivir ranges from 50 mg to 250 mg daily. In some embodiments, the effective amount of remdesivir ranges from 150 to 250 mg daily on day 1, and 50 mg to 150 mg daily on following days. In some embodiments, the method comprises administering intravenously remdesivir over a period of 5 to 10 days. In some embodiments, the human weighs from 3.5 kg to less than 40 kg and the effective amount of remdesivir ranges from 2.5 mg / kg to 5 mg / kg daily. In some embodiments, the effective amount of remdesivir ranges is 5 mg / kg daily on day 1, and 2.5 mg / kg daily on following days. In some embodiments, the method comprises administering intravenously remdesivir over a period of 5 to 10 days. In some embodiments, the method further comprise confirming if the human is infected with an Ebola virus prior to the administering. In some embodiments, the confirming is performed via a test that detects viral antigens or RNA in a sample of blood. In some embodiments, the confirming is performed via a test that detects viral antigens or RNA in a sample of bodily fluids other than blood.

[0161] Other aspects of the disclosure relate to a method of treating a human infected with or exposed to an Ebola virus, the method comprising administering to the human, for a suitable period of time, an effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino-phenylalanine- 4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3- hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir. In some embodiments, the pharmaceutically acceptable salt of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino- phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide is a hydrogen sulfate salt. In some embodiments, the effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof ranges from 200 mg to about 400 mg. In some embodiments, the effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxy carbonylpiperazide, free base or a pharmaceutically acceptable salt thereof is about 231 mg. In some embodiments, the effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino- phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof ranges from 50 mg to about 200 mg.

[0162] In some embodiments, remdesivir is administered intravenously. In some embodiments, the human weighs more than 40 kg and the effective amount of remdesivir ranges from 50 mg to 250 mg daily. In some embodiments, the effective amount of remdesivir ranges from 150 to 250 mg daily on day 1, and 50 mg to 150 mg daily on following days. In some embodiments, the method comprises administering intravenously remdesivir over a period of 5 to 10 days. In some embodiments, the human weighs from 3.5 kg to less than 40 kg and the effective amount of remdesivir ranges from 2.5 mg / kg to 5 mg / kg daily. In some embodiments, the effective amount of remdesivir ranges is 5 mg / kg daily on day 1, and 2.5 mg / kg daily on following days. In some embodiments, the method comprises administering intravenously remdesivir over a period of 5 to 10 days. In some embodiments, the method further comprises confirming if the human is infected with an Ebola virus prior to the administering. In some embodiments, the confirming is performed via a test that detects viral antigens or RNA in a sample of blood. In some embodiments, the confirming is performed via a test that detects viral antigens or RNA in a sample of bodily fluids other than blood.

[0163] Other aspects of the disclosure relate to a method of treating a human infected with or exposed to an Ebola virus, the method comprising administering to the human, for a suitable period of time, an effective amount of 3 -(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof, and an effective amount of N- a(2,4,6-triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof. In some embodiments, the effective amount of 3 -(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin- 4- ylmethyl)amide or a pharmaceutically acceptable salt thereof is administered orally. In some embodiments, the effective amount of 3-(4-chlorophenyl)- adamantane- 1-carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 9.5 mg / kg to about 15.5 mg / kg daily. In some embodiments, the effective amount of 3-(4- chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 10.5 mg / kg to about 14.5 mg / kg daily. In some embodiments, the effective amount of 3-(4-chlorophenyl)-adamantane- 1 -carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 11.5 mg / kg to about 13.5 mg / kg daily. In some embodiments, the effective amount of 3-(4- chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 15.0 mg / kg to about 20.0 mg / kg daily. In some embodiments, the method comprises administering orally a solid dosage form comprising the effective amount of the 3 -(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutically acceptable salt of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino- phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide is a hydrogen sulfate salt. In some embodiments, the effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof ranges from 200 mg to about 400 mg. In some embodiments, the effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof is about 231 mg. In some embodiments, the effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino- phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof ranges from 50 mg to about 200 mg. In some embodiments, the method further comprises confirming if the human is infected with an Ebola virus prior to the administering. In some embodiments, the confirming is performed via a test that detects viral antigens or RNA in a sample of blood. In some embodiments,

[0164] The confirming is performed via a test that detects viral antigens or RNA in a sample of bodily fluids other than blood.

[0165] Aspect of the disclosure relates to an active ingredient combination for treating Ebola virus infection comprising an effective amount of 3 -(4-chlorophenyl)-adamantane-1-carboxylic acid (pyri din-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir. In some embodiments, the combination is used to inhibit Ebola virus in a patient.

[0166] Other aspect of the disclosure relates to an active ingredient combination for treating Ebola virus infection comprising: an effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3- amidino-phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir. In some embodiments, the pharmaceutically acceptable salt of N-α-(2,4,6-triisopropylphenylsulfonyl)-3- hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide is a hydrogen sulfate salt. In some embodiments, the combination is used to inhibit Ebola virus in a patient.

[0167] Other aspect of the disclosure relates to an active ingredient combination for treating Ebola virus infection comprising: an effective amount of 3-(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof, and an effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy- carbonylpiperazide-hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino- phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof. In some embodiments, the combination is used to inhibit Ebola virus in a patient.

[0168] In an embodiment, a treatment for a ssRNA viral infection, such as Ebola virus further includes providing intravenous fluids (IV) and balancing electrolytes (body salts) to a subject being treated.

[0169] The compositions and methods of the disclosure can include formulation(s) of compound(s) that, upon administration to a subject, result in a concentration of the compound(s) that treats a filovirus-mediated disease. The compound(s) may be contained in any appropriate amount in any suitable carrier substance, and are generally present in an amount of 1-95% by weight of the total weight of the composition. The composition may be provided in a dosage form that is suitable for the oral, parenteral (e.g., intravenously or intramuscularly), rectal, dermatological, cutaneous, nasal, vaginal, inhalant, skin (patch), ocular, intrathecal, or intracranial administration route. Thus, the composition may be in the form of, e.g., tablets, capsules, pills, powders, granulates, suspensions, emulsions, solutions, gels including hydrogels, pastes, ointments, creams, plasters, drenches, osmotic delivery devices, suppositories, enemas, injectables, implants, sprays, or aerosols. The pharmaceutical compositions may be formulated according to conventional pharmaceutical practice.

[0170] Pharmaceutical compositions according to the disclosure or used in the methods of the disclosure may be formulated to release the active compound immediately upon administration or at any predetermined time or time period after administration. The latter types of compositions are generally known as controlled release formulations, which include (i) formulations that create substantially constant concentrations of the agent(s) of the disclosure within the body over an extended period of time; (ii) formulations that after a predetermined lag time create substantially constant concentrations of the agent(s) of the disclosure within the body over an extended period of time; (iii) formulations that sustain the agent(s) action during a predetermined time period by maintaining a relatively constant, effective level of the agent(s) in the body with concomitant minimization of undesirable side effects associated with fluctuations in the plasma level of the agent(s) (sawtooth kinetic pattern); (iv) formulations that localize action of agent(s), e.g., spatial placement of a controlled release composition adjacent to or in the diseased tissue or organ; (v) formulations that achieve convenience of dosing, e.g., administering the composition once per week or once every two weeks; and (vi) formulations that target the action of the agent(s) by using carriers or chemical derivatives to deliver the combination to a particular target cell type.

[0171] Any of a number of strategies can be pursued in order to obtain controlled release in which the rate of release outweighs the rate of metabolism of the compound in question. In one example, controlled release is obtained by appropriate selection of various formulation parameters and ingredients, including, e.g., various types of controlled release compositions and coatings. Thus, the compound(s) are formulated with appropriate excipients into a pharmaceutical composition that, upon administration, releases the compound(s) in a controlled manner. Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, molecular complexes, microspheres, nanoparticles, patches, and liposomes.

[0172] It is not intended that administration of compounds be limited to a single formulation and delivery method for all compounds of a combination. The combination can be administered using separate formulations and / or delivery methods for each compound of the combination using, for example, any of the above-described formulations and methods. In one example, a first agent is delivered orally, and a second agent is delivered intravenously. The dosage of a compound or a combination of compounds depends on several factors, including: the administration method, the type of disease to be treated, the severity of the infection, whether administration first occurs at an early or late stage of infection, and the age, weight, and health of the patient to be treated. For combinations that include a synergistic pair of agents identified herein, the recommended dosage for the anti-viral agent can be less than or equal to the recommended dose as given in the Physician's Desk Reference, 69thEdition (2015).

[0173] As described above, the compound(s) in question may be administered orally in the form of tablets, capsules, elixirs or syrups, or rectally in the form of suppositories. Parenteral administration of a compound is suitably performed, for example, in the form of saline solutions or with the compound(s) incorporated into liposomes. In cases where the compound in itself is not sufficiently soluble to be dissolved, a solubilizer such as ethanol can be applied. The correct dosage of a compound can be determined by examining the efficacy of the compound in viral replication assays, as well as its toxicity in humans.

[0174] EXAMPLES

[0175] The following examples are intended to illustrate rather than limit the disclosure.

[0176] Example 1- Opaganib and Upamostat Combinations with Remdesivir and Combinations of Opaganib and Upamostat Show Distinct Synergistic Effect Against Ebola

[0177] FIGS. 1-16 show that Opaganib and Upamostat demonstrated distinct synergistic effect when combined individually with remdesivir, significantly improving potency while maintaining cell viability, in an in vitro Ebola virus study.

[0178] Utilizing a checkerboard design to test the study compounds in combination, the study cell lines were pretreated and then infected with Ebola virus. The cells were fixed, washed and subjected to immunofluorescence staining using a virus-specific antibody as described below.

[0179] The antiviral activity of Opaganib was tested using image-based immunofluorescence screening assay. Dose response studies were performed to determine the potency (EC50), cytotoxicity (CC50) and selectivity index (SI) values. Opaganib was tested at a starting concentration of 60 μM, 3 -fold step dilutions, 8 different concentrations and four technical replicates. Cells were seeded one day prior to treatment and infection. After 20-24 hours, cells were pre-treated with the compound for 2 hours using HP-D300 digital dispenser. The cells were then infected with the virus using the optimized multiplicity of infection (MOI) and time of infection. At the end of the infection period, cells were fixed in 10% formalin. After a minimum of 24 hours or more of virus inactivation, plates were brought out of containment and subjected to standardized immunofluorescence staining assays using appropriate viral specific primary antibodies and appropriate fluorophore conjugated secondary antibodies. In addition, cell mask deep red and the Hoechst dyes were added to stain the cell cytoplasm and nuclei respectively. Images were acquired using the Opera Phenix confocal system and the percent infected cells and % loss in cell number were calculated directly by the Harmony software for each image with data normalized on a per plate basis. The quality of the assay for each plate was assessed using the Z’ (Z prime factor), which must be >0.5 for the assay to be acceptable. Analysis of dose response curves to determine the EC 50 was performed using GeneData Screener software applying the Levenberg-Marquardt algorithm (LMA) for curve fitting. The fitting strategy is considered acceptable if R2>0.8. Remdesivir was used as a reference inhibitor for each plate.

[0180] Ebola (EBOV), Marburg (MARV) and Sudan (SUDV) viruses are the three filoviruses which have caused the most fatalities in humans.

[0181] Table 1 : In vitro potency of Opaganib against EBOV, MARV and SUDV.

[0182] FIG. 11 shows the dose response curves to determine the potency (EC50), cytotoxicity (CC50) and selectivity index (SI) values. The upper curve shows the % activity of Opaganib in reducing cell number (EBOV / MRC-5 cells). The lower curve shows % activity of Opaganib in inhibiting viral infection. To determine if the combination of two drugs has synergistic or antagonistic activity, the data was analyzed using application, which incorporates four existing models (Loewe additivity model, Response additivity model, Highest single agent model (HSA), Bliss Independence Model, Zero Interaction Potency model (ZIP)) for drug combinations. Peak synergy is determined for all methods. A summary synergy score is generated and correspond to the average excess response due to drug interactions (i.e. synergy score of 15 corresponds to 15% of response beyond expectation). A score greater than 10 indicates synergy. The degree of combination synergy can be determined by comparing the observed drug combination response against the expected response, These four models quantify the degree of synergy either as the excess over the maximum single drug response (HSA), multiplicative effect of single drugs as if they acted independently (Bliss), expected response corresponding to an additive effect as if the single drugs were the same compound (Loewe), and expected response corresponding to the effect as if the single drugs did not affect the potency of each other (ZIP). A score from -10 to 10 indicates additivity. A score lower than -10 indicated antagonism.

[0183] The starting concentration of the drug varies in μM units with varying factor dilution. A mathematical equation was developed to calculate the % infection inhibition with no significant cell loss, and then these values were used to determine the synergy, antagonistic, additive, and potentiating effects of drug combinations.

[0184] In-vitro Combination (checkerboard) Assay set up

[0185] Different drug combinations were investigated to determine if the combination of the two drugs have synergistic or antagonist activity: Opaganib and Remdesivir (Example la), Opaganib and Remdesivir (Example lb) and Opaganib and Upamostat (a host protease inhibitor)(Example 1c). Upamostat inhibits cell surface proteases required to cleave viral proteins and thereby blocking attachment and uptake of the virus into host cells). MRC-5 cells were treated with individual drugs or combination of two drugs at an appropriate starting concentration. Exemplary Plate layout of the combination study set-up is shown in FIG. 1. A 3-fold serial dilution was performed, 8 different concentration and four technical replicates for each drug combination. In addition, each of the compounds were also individually tested at the same concentration in the same plate. After two hours of pre-treatment, the cells were infected with EBOV Duncan / Makona for 48 hours, fixed and subjected to immunofluorescence staining using viral antigen specific antibodies. In addition, cell mask deep red and the Hoechst dyes were added to stain the cell cytoplasm and nuclei respectively.

[0186] Images were acquired using the Opera Phenix confocal system and the percent infected cells and percent loss in cell number were calculated directly by the Harmony software for each image.

[0187] Example la. Opaganib Combinations with Remdesivir Show Distinct Synergistic Effect Against Ebola

[0188] Remdesivir was tested at starting concentration of 6 μM and Opaganib at starting concentration of 60 μM. Dose response involved three-fold serial dilution, eight different concentrations and four technical replicates either alone or in combination. See FIG. 2. Potency of individual drugs Opaganib and Remdesivir against EBOV is shown at Table 2 and FIG. 2.

[0189] Table 2

[0190] The raw data for the combination was analyzed to determine synergistic, additivity or antagonistic effects on viral inhibition while taking into account cell viability. FIG. 3 shows in vitro efficacy studies (% inhibition, % cell loss and % inhibition not due to cell death).

[0191] Four models are used to explore Synergistic effects: LOEWE, ZIP, BLISS and HSA. Peak synergy is determined for all methods.

[0192] Combination of host-directed Opaganib and Remdesivir, a direct acting antivirals exhibits synergistic activity. See FIG. 4 and FIG. 5, FIG. 12

[0193] Table 3 shows the highest significant peak synergy predicted by all four models (Loewe, HSA, Bliss and ZIP) is at a concentration of Opaganib at 0.74 μM and Remdesivir at 0.66 μM at 23% corrected inhibition. Table 3

[0194] Table 4 shows the highest significant peak synergy predicted by all four models (Loewe, HSA, Bliss and ZIP) is at a concentration of Opaganib at 0.74 μM and Remdesivir at 0.02 μM at 17% corrected inhibition.

[0195] Table 4 Example lb: Upamostat Combinations with Remdesivir Show Distinct Synergistic Effect Against Ebola

[0196] Upamostat is a first-in-class, once-daily orally administered investigational antiviral, that targets human serine proteases involved in preparing the spike protein for viral entry into target cells. Upamostat is well tolerated; in the initial COVID- 19 study, among 41 patients only one reported a drug-related adverse reaction (a mild, self-limited, rash).

[0197] Experimental design is shown in FIG. 6.

[0198] Dose Response curves of individual drugs.

[0199] Remdesivir was tested at starting concentration of 6 μM and Upamostat at starting concentration of 60 μM. Dose response involved three-fold serial dilution, eight different concentrations and four technical replicates either alone or in combination. See FIG. 7. Potency of individual drugs Upamostat and Remdesivir against EBOV is shown at FIG. 7.

[0200] FIG. 8 shows in vitro efficacy studies (%inhibition, % cell loss and % inhibition not due to cell death).

[0201] Combination of Upamostat and Remdesivir exhibits synergistic activity. See FIG. 5 and FIG. 9 and FIG. 10.

[0202] In vitro results for Opaganib and Upamostat show a distinct synergy in terms of viral inhibition while maintaining cell viability (i.e., not increasing toxicity), when either is added to remdesivir. Opaganib shows the greatest synergistic effect in combination with remdesivir.

[0203] The results suggest that Opaganib and Upamostat may be used in combination with direct antiviral agents, including but not limited as remdesivir, to improve treatment outcome, increasing efficacy while maintaining safety.

[0204] Opaganib (ABC294640) and Upamostat are both oral, host-directed, small molecule investigational drugs that are easy to administer and distribute, with demonstrated activity against multiple viral targets, including COVID-19, and are expected to be effective against emerging viral variants. This, together with their growing safety and tolerability databases, presents a compelling hypothesis for treatment of Ebola virus.

[0205] Twice daily administered Opaganib has previously demonstrated benefit in late-stage clinical studies of patients hospitalized with moderate to severe COVID-19. Opaganib is believed to be the first host-directed molecule to show activity in Ebola virus disease, having recently delivered a statistically significant increase in survival time in an in vivo Ebola virus study.

[0206] Opaganib, a host-directed and potentially broad-acting drug, is a first-in-class, orally administered sphingosine kinase-2 (SPHK2) selective inhibitor with anticancer, anti-inflammatory and antiviral activity.

[0207] Opaganib ’s host-directed action is thought to work through the inhibition of multiple pathways, the induction of autophagy and apoptosis, and disruption of viral replication, through simultaneous inhibition of three sphingolipid-metabolizing enzymes in human cells (SPHK2, DES1 and GCS).

[0208] Opaganib has demonstrated antiviral activity against SARS-CoV-2, multiple variants, and several other viruses, such as Influenza A. Being host-targeted, and based on data accumulated to date, Opaganib is expected to maintain effect against emerging viral variants. In prespecified analyses of Phase 2 / 3 clinical data in hospitalized patients with moderate to severe COVID-19, oral Opaganib demonstrated improved viral RNA clearance, faster time to recovery and significant mortality reduction in key patient subpopulations versus placebo on top of standard of care.

[0209] Example 1c- Opaganib and Upamostat Combinations Show Distinct Synergistic Effect Against Ebola Upamostat and Opaganib were tested at starting concentration of 60 μM. Dose response involved three-fold serial dilution, eight different concentrations and four technical replicates either alone or in combination. This combination study was performed in HeLa cells.

[0210] Dose response curves of individual drugs are shown in FIG. 13.

[0211] Potency of individual drugs is shown at Table 5.

[0212] Table 5 FIG. 14 and FIG. 15 show the topology synergy maps. Table 6 shows the highest significant peak synergy predicted by all four models (Loewe, HSA, Bliss and ZIP) is at a concentration of

[0213] Opaganib at 20 μM and Upamostat at 6.6 μM with 98 % corrected inhibition.

[0214] Table 6

[0215] Table 7 shows the highest significant peak synergy predicted by all four models (Loewe, HSA, Bliss and ZIP) is at a concentration of Opaganib at 2.2 μM and Upamostat at 6.6 μM with 46-63 % inhibition.

[0216] Table 7

[0217] Example 2: Evaluation of the efficacy of Opaganib in mouse model of EBOV infection

[0218] Two independent studies were performed to determine the efficacy of Opaganib in a mouse model of EBOV infection. In the first study, Balb / c mice (6-8 weeks old; n=10 mice / group) were exposed via the intraperitoneal (i.p) route with 100 plaque forming units (pfu) of mouse-adapted ebola virus. (maEBOV). Approximately one-hour post-infection, treatment with different doses of Opaganib (60 mg / kg, 100 mg / kg and 150 mg / kg) was initiated. Doses were chosen based on the lowest dose that demonstrated efficacy in any of our mouse models. The dose close to the Human Equivalent Dose at 500mg BID selected for our clinical trials and at the likely Maximum Tolerated Human Equivalent Dose, respectively. Mice administered with vehicle formulation (46.7% PEG400, 46.7% Saline and 6.6% Tween 80) served as the infection control group. Both Opaganib and the vehicle control were administered via oral route and given twice daily (BID) for total of 8 days. Favipiravir (150 mg / kg) was used as a reference control and was administered via the oral route, twice daily for 8 days. Mice were observed daily for signs of disease and were euthanized when moribund. Animals that survived until the end of the study were euthanized on day 23. The data showed statistically significant increase in survival at Opaganib 150 mg / kg BID in the mouse model:

[0219] - 30% survival in the 150 mg / kg Opaganib treated group compared to no survival in the vehicle control (p<0.03);

[0220] - Virus exposure was 100% lethal to vehicle-control mice.

[0221] In the second serial sac study, Balb / c mice were exposed via the intraperitoneal (i.p.) route with 100 pfu of maEBOV. Approximately one-hour post-infection, mice (n=10 mice / group) were administered twice daily via the oral route with Opaganib (150 mg / kg) or vehicle control for a total of 8 days. These mice were monitored for survival until the end of study. A cohort of mice (n=3 mice / timepoint) were euthanized on days 5, 7 and 10 to determine viremia in blood and tissues. Figure 9 shows the combined survival curve generated from two independent studies performed for Vehicle control and Opaganib (150 mg / kg). Mice treated with Opaganib (150 mg / kg) overall showed a 20% survival compared to the vehicle control which showed 10% survival. See FIG. 16.

[0222] All patents, patent applications, and published references cited herein are hereby incorporated by reference in their entirety. Various modifications and variations of the described compositions and methods of the disclosure will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific embodiments, it will be understood that the invention should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention that are obvious to those skilled in the fields of molecular biology, medicine, immunology, pharmacology, virology, or related fields are intended to be within the scope of the disclosure.

Claims

CLAIMS1. A method of treating a human infected with or exposed to an Ebola virus, the method comprising: administering to the human, for a suitable period of time, an effective amount of 3-(4- chlorophenyl)-adamantane-1-carboxylic acid (pyri din-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir.

2. A method of treating a human infected with or exposed to an Ebola virus, the method comprising: administering to the human, for a suitable period of time, an effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir.

3. A method of treating a human infected with or exposed to an Ebola virus, the method comprising: administering to the human, for a suitable period of time, an effective amount of 3-(4- chlorophenyl)-adamantane-1-carboxylic acid (pyri din-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof, and an effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3- amidino-phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof.

4. The method of claim 1 or claim 3 wherein the effective amount of 3-(4-chlorophenyl)- adamantane-1 -carboxylic acid (pyri din-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof is administered orally.

5. The method of claim 4, wherein the effective amount of 3-(4-chlorophenyl)- adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 9.5 mg / kg to about 15.5 mg / kg daily.

6. The method of claim 4, wherein the effective amount of 3-(4-chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 10.5 mg / kg to about 14.5 mg / kg daily.

7. The method of claim 4, wherein the effective amount of 3-(4-chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 11.5 mg / kg to about 13.5 mg / kg daily.

8. The method of claim 4, wherein the effective amount of 3-(4-chlorophenyl)-adamantane-1- carboxylic acid (pyridin-4-ylmethyl)amide or a pharmaceutically acceptable salt thereof ranges from about 15.0 mg / kg to about 20.0 mg / kg daily.

9. The method of any one of claims 1-7, comprising administering orally a solid dosage form comprising the effective amount of the 3 -(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof.

10. The method of claim 1 or claim 2 wherein remdesivir is administered intravenously.

11. The method of claim 10, wherein the human weighs more than 40 kg.

12. The method of claim 11, wherein the effective amount of remdesivir ranges from 50 mg to 250 mg daily.

13. The method of claim 11, wherein the effective amount of remdesivir ranges from 150 to 250 mg daily on day 1, and 50 mg to 150 mg daily on following days.

14. The method of claim 13, comprising administering intravenously remdesivir over a period of 5 to 10 days.

15. The method of claim 10, wherein the human weighs from 3.5 kg to less than 40 kg.

16. The method of claim 15, wherein the effective amount of remdesivir ranges from 2.5 mg / kg to 5 mg / kg daily.

17. The method of claim 15, wherein the effective amount of remdesivir ranges is 5 mg / kg daily on day 1, and 2.5 mg / kg daily on following days.

18. The method of claim 17, comprising administering intravenously remdesivir over a period of 5 to 10 days.

19. The method of claim 1 or claim 3, wherein the pharmaceutically acceptable salt of N-α-(2,4,6- triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4-ethoxycarbonylpiperazide is a hydrogen sulfate salt.

20. The method of claim 2 or claim 3, wherein the effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxy carbonylpiperazide, free base or a pharmaceutically acceptable salt thereof ranges from 200 mg to about 400 mg.

21. The method of claim 20, wherein the effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxy carbonylpiperazide, free base or a pharmaceutically acceptable salt thereof is about 231 mg.

22. The method of claim 2 or claim 3, wherein the effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide- hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof ranges from 50 mg to about 200 mg.

23. The method of claim any preceding claims, comprising confirming if the human is infected with an Ebola virus prior to the administering.

24. The method of claim 23, wherein the confirming is performed via a test that detects viral antigens or RNA in a sample of blood.

25. The method of claim 23, wherein the confirming is performed via a test that detects viral antigens or RNA in a sample of bodily fluids other than blood.

26. An active ingredient combination for treating Ebola virus infection comprising: an effective amount of 3 -(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir.

27. An active ingredient combination for treating Ebola virus infection comprising: an effective amount of N-α(2,4,6-triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy- carbonylpiperazide-hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino- phenylalanine-4-ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof, and an effective amount of remdesivir.

28. An active ingredient combination for treating Ebola virus infection comprising: an effective amount of 3-(4-chlorophenyl)-adamantane-1-carboxylic acid (pyri din-4- ylmethyl)amide or a pharmaceutically acceptable salt thereof, and an effective amount of N-α(2,4,6- triisopropylphenylsulfonyl)-3-amidino-phenylalanine-4-ethoxy-carbonylpiperazide-hydrochloride, N-α-(2,4,6-triisopropylphenylsulfonyl)-3-hydroxyamidino-phenylalanine-4- ethoxycarbonylpiperazide, free base or a pharmaceutically acceptable salt thereof.

29. The combination of any one of claims 26-28, wherein the combination is for use in inhibiting Ebola virus in a patient.

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