HK1237643B

HK1237643B - Therapy for inhibition of single-stranded rna virus replication

Info

Publication number: HK1237643B
Application number: HK17110471.8A
Authority: HK
Inventors: 列扎‧法西; 德罗尔‧本-阿舍; 盖伊‧戈德堡
Original assignee: 红山生物医药有限公司
Priority date: 2014-10-24
Filing date: 2015-10-23
Publication date: 2021-10-08

Abstract

Pharmaceutical compositions showing the ability to inhibit or suppress replication of a filovirus in an individual are disclosed. The disclosed compositions are useful for treating, reventing, or reducing the spread of infections by filovirus. A method includes administering at least one agent of the present disclosure to an individual infected with or exposed to a filovirus, wherein the step of administering is carried out for a suitable time period so that the individual is treated; and determining whether the individual has been treated, wherein the step of determining includes one of measuring an inhibition in viral replication, measuring a decrease in viral load, or reducing at least one symptom associated with the filovirus.

Description

Treatment for inhibiting the replication of single-stranded RNA viruses

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of and priority to U.S. Provisional Patent Application No. 62/068,465, filed on October 24, 2014, U.S. Provisional Patent Application No. 62/068,469, filed on October 24, 2014, U.S. Provisional Patent Application No. 62/068,477, filed on October 24, 2014, U.S. Provisional Patent Application No. 62/068,487, filed on October 24, 2014, U.S. Provisional Patent Application No. 62/068,492, filed on October 24, 2014, and U.S. Provisional Patent Application No. 62/188,030, filed on July 2, 2015, the entire contents of which are incorporated herein by reference.

Background Art

RNA viruses can be divided into negative-sense (-) RNA viruses and positive-sense (+) RNA viruses based on the sense or polarity of their RNA translation. The largest family of viruses is the single-stranded negative-sense (-) RNA ("ssRNA") virus family. Their viral RNA genome cannot be directly translated, but the (-) strand is complementary to the viral mRNA that needs to be produced and translated into viral proteins. At the time of this disclosure, one order and eight families have been recognized in this group. There are also some unclassified species and genera:

Single-molecule negative-strand RNA viruses

○ Bornaviridae family - Borna disease virus

○ Filoviridae family - includes Ebola virus, Marburg virus

Paramyxoviridae family - includes measles virus, mumps virus, Nipah virus, Hendra virus, respiratory tract infection virus (RSV), and Newcastle disease virus (NDV)

○ Rhabdoviridae family - includes rabies virus

Nyamiviridae family - includes niaviruses

Unclassified family:

Arenaviridae family – includes Lassa virus

Bunyaviridae family - includes Hantavirus, Crimean-Congo hemorrhagic fever

○Ophioviridae family

○ Orthomyxoviridae family - includes influenza viruses

Unclassified genus:

○Hepatitis D virus genus - including hepatitis E virus

○Dichlorovirus

○Emaravirus

Nivavirus – includes Niamani virus and Midway virus

○Tensivirus

○Meganevirus

Unclassified species:

Taastrup virus

○ Sclerotinia sclerotiorum negative-strand RNA virus 1

Despite decades of research efforts to develop an effective, approved, and available filovirus treatment for individuals, there are currently no U.S. Food and Drug Administration-approved vaccines or therapies for the treatment of filovirus disease infections.

Summary of the Invention

Disclosed herein are therapies for inhibiting the replication of single-stranded RNA viruses. Disclosed herein are compositions and methods for treating symptomatic and/or asymptomatic infection with negative-sense single-stranded RNA viruses, including but not limited to, Boyaviridae, Filoviridae, Paramyxoviridae, Rhabdoviridae, Tobaccoviridae, or any combination thereof. In one embodiment, disclosed herein are therapies for inhibiting Ebola virus. In one embodiment, disclosed herein are therapies for inhibiting Marburg virus.

The methods of the present invention comprise administering a compound to an individual infected with or exposed to a filovirus, wherein the administering step is performed for a suitable period of time such that the individual is treated, and wherein the compound is represented by Formula I:

or a pharmaceutically acceptable salt thereof, wherein:

_R1 is phenyl, 4-chlorophenyl or 4-fluorophenyl,

R ₂ is 4-pyridyl, optionally substituted with up to 4 groups which are independently (C ₁ -C ₆ ) alkyl, halogen, haloalkyl, -OC(O)(C ₁ -C ₆ alkyl), -C(O)O(C ₁ -C ₆ alkyl), -CONR'R", -OC(O)NR'R", -NR'C(O)R", -CF ₃ , -OCF ₃ , -OH, C ₁ -C ₆ alkoxy, hydroxyalkyl, -CN, -CO ₂ H, -SH, -S-alkyl, -SOR'R", -SO ₂ R', -NO ₂ or NR'R", wherein R' and R" are independently H or (C ₁ -C ₆ ) alkyl, and wherein each alkyl portion of the substituent is optionally further substituted with 1, 2 or 3 groups independently selected from halogen, CN, OH and NH ₂ ,

_R4 is H or alkyl, and

n is 1 or 2; and

Determining whether the individual has been treated, wherein the determining step comprises the step of measuring one of: measuring inhibition of viral replication, measuring a reduction in viral load, or alleviating at least one symptom associated with the filovirus. In one embodiment, the compound of formula I is:

In one embodiment, the compound of Formula I is administered at a daily dose of about 2.5 mg/kg to about 22.5 mg/kg. In one embodiment, the compound of Formula I is administered at a daily dose of about 3.5 mg/kg to about 21.5 mg/kg. In one embodiment, the compound of Formula I is administered at a daily dose of about 4.5 mg/kg to about 20.5 mg/kg. In one embodiment, the compound of Formula I is administered at a daily dose of about 5.5 mg/kg to about 19.5 mg/kg. In one embodiment, the compound of Formula I is administered at a daily dose of about 6.5 mg/kg to about 18.5 mg/kg. In one embodiment, the compound of Formula I is administered at a daily dose of about 7.5 mg/kg to about 17.5 mg/kg. In one embodiment, the compound of Formula I is administered at a daily dose of about 8.5 mg/kg to about 16.5 mg/kg. In one embodiment, the compound of Formula I is administered at a daily dose of about 9.5 mg/kg to about 15.5 mg/kg. In one embodiment, the compound of Formula I is administered at a daily dose of about 10.5 mg/kg to about 14.5 mg/kg. In one embodiment, the compound of formula I is administered at a daily dose of about 11.5 mg/kg to about 13.5 mg/kg. In one embodiment, the determining step includes measuring viral load using a nucleic acid amplification-based test method at at least two different times within a suitable time period. In one embodiment, the inhibition of viral replication or the reduction of viral load determined using a nucleic acid amplification-based test method is at least 10%. In one embodiment, the individual is a human. In one embodiment, the filovirus is Ebola virus or Marburg virus. In one embodiment, the filovirus is Ebola virus. In one embodiment, the compound of formula I is present in a solid dosage form. In one embodiment, the solid dosage form is a capsule. In one embodiment, the method further includes administering at least one antibiotic to an individual infected or exposed to a filovirus for a suitable time period, wherein the combination of at least one antibiotic and the compound of formula I produces a synergistic effect. In one embodiment, the at least one antibiotic is selected from one of clarithromycin or rifabutin.

The methods of the present invention comprise administering at least two antibiotics to an individual infected with or exposed to a filovirus, wherein the administering step is performed for a suitable period of time to allow the individual to be treated; and determining whether the individual has been treated, wherein the determining step comprises a step of measuring one of: measuring inhibition of viral replication, measuring a reduction in viral load, or alleviating at least one symptom associated with the filovirus. In one embodiment, at least one antibiotic is a macrolide antibiotic. In one embodiment, at least one antibiotic is a rifamycin antibiotic. In one embodiment, the antibiotics are clarithromycin and rifabutin. In one embodiment, clarithromycin is administered at a daily dose of about 2.5 mg/kg to about 21.5 mg/kg. In one embodiment, clarithromycin is administered at a daily dose of about 3.5 mg/kg to about 20.5 mg/kg. In one embodiment, clarithromycin is administered at a daily dose of about 4.5 mg/kg to about 19.5 mg/kg. In one embodiment, clarithromycin is administered at a daily dose of about 5.5 mg/kg to about 18.5 mg/kg. In one embodiment, clarithromycin is administered at a daily dose of about 6.5 mg/kg to about 17.5 mg/kg. In one embodiment, rifabutin is administered at a daily dose of about 7.5 mg/kg to about 16.5 mg/kg. In one embodiment, clarithromycin is administered at a daily dose of about 8.5 mg/kg to about 15.5 mg/kg. In one embodiment, clarithromycin is administered at a daily dose of about 9.5 mg/kg to about 14.5 mg/kg. In one embodiment, rifabutin is administered at a daily dose of about 10.5 mg/kg to about 13.5 mg/kg. In one embodiment, rifabutin is administered at a daily dose of about 0.5 mg/kg to about 7.5 mg/kg. In one embodiment, rifabutin is administered at a daily dose of about 1.5 mg/kg to about 6.5 mg/kg. In one embodiment, rifabutin is administered at a daily dose of about 2.5 mg/kg to about 5.5 mg/kg. In one embodiment, the determining step comprises measuring viral load using a nucleic acid amplification-based assay at at least two different times within a suitable time period. In one embodiment, the inhibition of viral replication or reduction in viral load as determined using a suitable test is at least 10%. In one embodiment, the individual is a human. In one embodiment, the filovirus is an Ebola virus or a Marburg virus. In one embodiment, the filovirus is an Ebola virus.

DETAILED DESCRIPTION

As used herein, the term "pharmaceutical agent" refers to a compound that has pharmacological activity - the effect of the agent on a subject. The terms "pharmaceutical agent," "compound," and "drug" are used interchangeably herein.

"Patient" or "subject" refers to any animal, such as a primate. In one embodiment, the primate is a non-human primate. In one embodiment, the primate is a human primate. Any animal can be treated using the methods and compositions of the present invention.

As used herein, the term "synergistic effect" refers to the coordinated or interrelated effects of two or more agents of the present invention such that the combined effect is greater than the sum of the effects of each agent alone. In one embodiment, when administered together as part of a therapeutic regimen, the agents of the present invention provide therapeutic synergy without the attendant side effects of the synergy (such as, but not limited to, cross-reacting agents).

As used herein, the term "treatment" refers to the administration of one or more agents of the present invention to moderately inhibit viral replication in vitro or in vivo, to moderately reduce the load of viruses in cells in vitro or in vivo, or to reduce at least one symptom associated with a virally mediated disease. Ideally, the inhibition of viral replication or the reduction of viral load as measured using a suitable test is at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%. Assays for monitoring viral replication include, but are not limited to, cytopathic virus assays, reporter virus and reporter cell assays, viral replicon assays, and gene-targeted viral assays. In one embodiment, the slowing or stopping of viral replication is measured using an assay that measures the inhibition of viral replication mediated by CD8 T cells. Viral load testing can be performed using nucleic acid amplification-based tests (NAT or NAAT) and non-nucleic acid-based tests on plasma samples to determine the amount of virus in a given volume, including viral RNA levels and total viral DNA in plasma and tissues. Alternatively, in certain embodiments, treatment observed by trained physicians is a considerable or substantial alleviation of symptoms in patients with virally mediated diseases. Typically, reduction in viral replication is achieved by reducing RNA polymerization rates, RNA translation rates, protein processing or modification rates, or by reducing the activity of molecules involved in any viral replication step (e.g., proteins or genome encoding of viruses or hosts that are important for viral replication). In one embodiment, the term "treatment" refers to the ability of an agent of the present invention to inhibit or suppress the replication of a virus such as an RNA virus. In one embodiment, the term "treatment" refers to the ability of an agent of the present invention or an agent to inhibit cytopathic effects during RNA viral infection.

"Effective amount" refers to the amount (such as any virus described herein, including Ebola virus or Marburg virus) required for the treatment of a patient with a viral disease by the medicament of the present invention alone or in combination with other treatment regimens in a clinically relevant manner. The sufficient effective amount of the medicament for treating the illness caused by a virus for the implementation of the present invention varies according to the mode of administration, the age of the patient, the weight of the patient, and the general health status of the patient. Ultimately, the prescriber will determine the appropriate dosage and dosage regimen. In the combination therapy of the present invention, if the reagent is administered in a non-combination (single agent) treatment manner, the effective amount of the medicament may be less than the effective amount. In addition, the effective amount can be the amount of the medicament in the combination therapy of the present invention, which is safe and effective (such as the U.S. Food and Drug Administration) in treating patients with viral diseases relative to each separate medicament determined and approved by regulatory agencies.

"More effective" means that a treatment has greater efficacy, or is less toxic, safer, more convenient, or less expensive than the other treatment to which the treatment is being compared. Efficacy can be measured by a skilled practitioner using any standard method appropriate for a given indication.

"Filovirus" refers to a virus belonging to the family Fillovirdae. Exemplary filoviruses are Ebola virus and Marburg virus.

"Ebola" or "Ebola hemorrhagic fever" is a disease caused by infection with one of the Ebola virus strains. Ebola viruses can cause disease in humans and non-human primates (monkeys, gorillas, and chimpanzees). Ebola disease in humans is caused by four of five viruses in the genus Ebolavirus. These four viruses are Bundibugyou ebolavirus (BDBV), Sudan virus (SUDV), Côte d'Ivoire ebolavirus (TAFV), and another simply called Ebolavirus (EBOV, formerly known as Zaire ebolavirus). The fifth virus, Reston virus (RESTV), is not thought to cause disease in humans but does cause disease in other primates. These five viruses are closely related to Marburg virus. Marburg virus disease (MVD) is a serious disease in humans and non-human primates caused by one of two Marburg viruses (Marburg virus and Raf virus).

As used herein, the term "suitable period of time" refers to the period of time that begins when a patient diagnosed with an ssRNA viral infection (such as, but not limited to, Ebola) is treated using the methods of the present disclosure, including the entire treatment period, until the patient discontinues treatment due to a decrease in symptoms associated with the ssRNA viral infection (such as, but not limited to, Ebola) or due to laboratory diagnosis indicating that the ssRNA viral infection (such as, but not limited to, Ebola) is under control. In one embodiment, the suitable period of time is one (1) week. In one embodiment, the suitable period of time is between one (1) week and two (2) weeks. In one embodiment, the suitable period of time is two (2) weeks. In one embodiment, the suitable period of time is between two (2) weeks and three (3) weeks. In one embodiment, the suitable period of time is three (3) weeks. In one embodiment, the suitable period of time is between three (3) weeks and four (4) weeks. In one embodiment, the suitable period of time is four (4) weeks. In one embodiment, the suitable period of time is between four (4) weeks and five (5) weeks. In one embodiment, the suitable period of time is five (5) weeks. In one embodiment, a suitable time period is between five (5) weeks and six (6) weeks. In one embodiment, a suitable time period is six (6) weeks. In one embodiment, a suitable time period is between six (6) weeks and seven (7) weeks. In one embodiment, a suitable time period is seven (7) weeks. In one embodiment, a suitable time period is between seven (7) weeks and eight (8) weeks. In one embodiment, a suitable time period is eight (8) weeks.

As used herein, the term "cytopathic effect" refers to changes in cell morphology caused by viral infection.

As used herein, the term "cellular pathogenesis" or "pathogenesis" includes inhibition of host cell gene expression and includes other cellular changes beyond those visible at the microscopic level that contribute to viral pathogenesis.

As used herein, the term "in vitro" refers to a procedure performed in an artificial environment, such as, but not limited to, a test tube or a cell culture system. Those skilled in the art will appreciate that, for example, an isolated SK enzyme can be contacted with a modulator in an in vitro environment. Alternatively, isolated cells can be contacted with a modulator in an in vitro environment.

As used herein, the term "in vivo" refers to procedures performed within a living organism, such as, but not limited to, humans, monkeys, mice, rats, rabbits, cows, horses, pigs, dogs, felines, or primates.

Some small viruses carry their genomes as single-stranded DNA (ssDNA) molecules. These viruses have simple genomes: one gene for the viral nucleocapsid protein and another for the DNA replicase. Viruses with ssDNA genomes also face severe replication problems in host cells. When introduced into cells, these genomes cannot be used to produce viral proteins because the only transcription template is double-stranded DNA. Therefore, the first step after infection is to convert the viral ssDNA into dsDNA using host cell DNA polymerases. In some of these viruses, the 3' end of the viral DNA folds over and forms dsDNA through base pairing with internal sequences. In this way, primers are built into the genome, and the 3' end can be amplified to produce dsDNA that serves as a transcription template. The resulting transcripts are translated into viral proteins, the replicated viral DNA is converted back into the ssDNA genome, and the virions are packaged for export.

RNA viruses can be divided into negative-sense (-) and positive-sense (+) or bidirectional RNA viruses according to the translation sense or polarity of their RNA. Positive-sense viral RNA is similar to mRNA and can therefore be translated immediately by the host cell. Negative-sense viral RNA is complementary to mRNA and therefore must be converted into positive-sense RNA by RNA polymerase before translation. Therefore, purified RNA of positive-sense viruses can directly cause infection, although it may be less infectious than whole virus particles. Purified RNA of negative-sense viruses is not infectious in itself because it needs to be transcribed into positive-sense RNA; each virus particle can be transcribed into several positive-sense RNAs. Bidirectional RNA viruses are similar to negative-sense RNA viruses, except that they also convert genes from the positive strand. Examples of positive-strand RNA viruses include, but are not limited to, poliovirus, coxsackievirus, and echovirus. Examples of negative-strand RNA viruses include, but are not limited to, influenza virus, measles virus, and rabies virus.

The largest family of viruses is the (-)ssRNA virus family. Their viral RNA genomes cannot be directly translated, but rather the (-) strand complements the viral mRNA required for the production and translation of viral proteins. Hundreds of different (-)ssRNA viruses have been found in nature, ranging from measles and influenza viruses to rabies and Ebola. This group of viruses includes members of the Ebola and influenza virus families.

Ebola, formerly known as Ebola hemorrhagic fever, is a disease caused by infection with a strain of the Ebola virus. Ebola viruses can cause disease in humans and non-human primates (monkeys, gorillas, and chimpanzees). Ebola disease in humans is caused by four of five species in the genus Ebolavirus. These four viruses are Bundibugyou ebolavirus (BDBV), Sudan virus (SUDV), Côte d'Ivoire ebolavirus (TAFV), and a species simply called Ebolavirus (EBOV, formerly known as Zaire ebolavirus). The fifth virus, Reston virus (RESTV), is not thought to cause disease in humans but does cause disease in other primates. These five viruses are closely related to Marburg virus. Currently, there are no specific treatments proven to be effective for Ebola.

Ebola virus contains a single-stranded, non-infectious RNA genome. The Ebola virus genome is approximately 19 kilobase pairs long and contains seven genes with the sequence 3'-UTR-NP-VP35-VP40-GP-VP30-VP24-L-5'-UTR. The genomes of the five different Ebola viruses (BDBV, EBOV, RESTV, SUDV, and TAFV) differ in the amount and position of sequence and gene overlap. In general, Ebola viruses are 80 nanometers (nm) wide and can be up to 14,000 nm long. Typically, the median particle length of Ebola virus ranges from 974 to 1,086 nm (in contrast to Marburg virus, where the median particle length is measured at 795-828 nm), but particles up to 14,000 nm have been detected in tissue culture.

Viral matrix protein 40 (VP40) is the most abundant protein in virions, infected cells, and within the viral nucleocapsid. The nucleoprotein (NP) associates with the viral genome and, together with the polymerase cofactor (VP35), the transcriptional activator (VP30), and the RNA-dependent RNA polymerase (L), assembles into a helical nucleocapsid (NC). The viral proteins that comprise the NC catalyze the replication and transcription of the viral genome. NC assembly also requires a minor viral matrix protein, VP24. If NP is expressed alone in cells, it forms a loose coil-like structure with cellular RNA. 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. VP24 and the viral matrix protein VP40 have been shown to reduce the efficiency of transcription and replication of the EBOV genome, suggesting that VP24 and VP40 are important for the conversion of the transcription- and replication-competent NC to one that is ready for viral assembly. Even when expressed alone, VP40 plays a role in the formation and release of enveloped, filamentous virus-like particles (VLPs). When VP40 was co-expressed with NP, VP35, and VP24, NC-like structures were incorporated into VLPs, indicating that direct interaction between VP40 and NP is important for recruiting NC-like structures to the plasma membrane at the site of budding. The interaction between VP40 and NP is also required for the formation of condensed NC-like structures. GP is a surface glycoprotein that forms a spike on the virion and plays a key role in viral entry into cells by mediating receptor binding and fusion.

The Ebola virus life cycle begins with the attachment of the virion to a specific cell surface receptor, followed by fusion of the virion membrane with the cell membrane and simultaneous release of the viral nucleocapsid into the cytosol. Ebola virus structural glycoproteins (called GP1 and GP2) are responsible for the virus's ability to bind to and infect target cells. The viral RNA polymerase, encoded by the L gene, partially unmasks the nucleocapsid and transcribes the gene into positive-strand mRNA, which is then converted into structural and nonstructural proteins. The most abundant protein produced is the nucleoprotein, and its cellular concentration determines the timing of the transition from L gene transcription to genome replication. Replication produces full-length positive-strand antigen monomers, which are in turn transcribed into negative-strand viral progeny genome copies. The newly synthesized structural proteins and genome self-assemble and accumulate near the cell membrane. Viruses shed from the cell, acquiring their envelope from the cell membrane from which they budded. Mature progeny particles then infect other cells, repeating the cycle. Ebola virus genetics are difficult to study due to its toxicity.

Ebola virus is a filamentous, enveloped, negative-sense RNA virus in the Filoviridae family. The RNA-dependent RNA polymerase of Ebola virus shares significant sequence homology with other negative-sense RNA viruses, required for viral transcription and replication of the viral genome. However, the RNA-dependent RNA polymerase requires both host factors and viral proteins to coordinate replication and transcription. One reason Ebola virus is so deadly is its ability to evade the immune system while actively destroying the body, thereby preventing the immune system from mustering a concerted effort to combat the disease. During infection, monocytes/macrophages in lymphoid tissues are early and persistent targets of the deadly virus. During 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 clinical features observed in Ebola hemorrhagic fever. Secreted cytokines, chemokines, and other mediators alter vascular function, promoting and recruiting the influx of inflammatory cells, including additional monocytes/macrophages, to the site of infection. Viruses released from infected macrophages and dendritic cells spread throughout similar cells throughout the body and to parenchymal cells in many organs, leading to multifocal tissue necrosis. The host's ability to mount an effective adaptive immune response is impaired by massive lymphocyte apoptosis, a phenomenon also seen in bacterial sepsis. Ebola infection also induces lymphocyte apoptosis, despite the virus being unable to replicate in lymphocytes. Recent studies have shown that NK (natural killer) cells and CD4+ and CD8+ lymphocytes are the primary cell types affected in Ebola-infected macaques.

Ebola subjects who show symptoms (i.e., are symptomatic) may undergo blood and/or tissue tests to confirm a diagnosis of Ebola, including but not limited to high fever, headache, joint and muscle aches, sore throat, weakness, stomach pain, lethargy, and loss of appetite. Ebola subjects who do not show symptoms (i.e., are asymptomatic) may undergo blood and/or tissue tests to confirm a diagnosis of Ebola. These asymptomatic subjects may have markers in their blood that indicate they are carrying the disease, but they are completely asymptomatic.

Subjects infected with ssRNA viruses that develop symptoms (i.e., are symptomatic) may undergo blood and/or tissue tests to confirm the diagnosis of ssRNA virus infection, including but not limited to high fever, headache, joint and muscle aches, sore throat, weakness, stomach pain, lethargy, and loss of appetite. Subjects infected with ssRNA viruses that do not show symptoms (i.e., are asymptomatic) may undergo blood and/or tissue tests to confirm the diagnosis of ssRNA virus infection. These asymptomatic subjects may have markers in their blood that indicate they are carrying the disease, but they are completely asymptomatic.

In one embodiment, viral antigens or RNA can be detected in blood and other body fluids within a few days after the onset of symptoms or after treatment according to the present disclosure by collecting a sample of blood or other body fluid and testing the sample using methods such as antigen capture enzyme-linked immunosorbent assay (ELISA), IgM ELISA (to determine whether the subject has IgM antibodies), IgG ELISA (to determine whether the subject has IgG antibodies), polymerase chain reaction (PCR), or by virus isolation.

The present disclosure identifies agents and combinations of agents that have activity in inhibiting model filoviruses.The invention features compositions and methods for treating diseases mediated by filoviruses, such as those caused by Ebola virus or Marburg virus.

In one embodiment, the present disclosure describes a method for treating a patient suffering from a filovirus-mediated disease, such as caused by Ebola virus or Marburg virus. The method comprises administering to the patient a first agent selected from the group consisting of agents listed in Table 1 or an analog thereof, in an amount effective to treat the patient. In one embodiment, the method further comprises administering a second agent selected from the group consisting of agents listed in Table 1. In one embodiment, the method further comprises administering a third agent selected from the group consisting of agents listed in Table 1.

Table 1

When the method comprises administering more than one active agent to a patient, the agents can be administered within 7, 6, 5, 4, 3, 2, or 1 day; within 24, 12, 6, 5, 4, 3, 2, or 1 hour; within 60, 50, 40, 30, 20, 10, 5, or 1 minute; or substantially simultaneously. The methods of the invention may comprise administering one or more agents to a patient orally, systemically, parenterally, topically, intravenously, by inhalation, or intramuscularly. In one embodiment, the methods of the invention comprise administering one or more agents to a patient orally.

In one embodiment, the present disclosure describes a composition comprising two or more agents selected from the group consisting of agents listed in Table 1. In one embodiment, the two or more agents are present in amounts that, when administered together, are effective to treat a patient suffering from a filovirus-mediated disease, such as a disease caused by Ebola virus or Marburg virus. In one embodiment, the composition consists of an active ingredient consisting of two or more agents selected from the group consisting of agents listed in Table 1 and an excipient.

Active ingredients or agents useful in the present invention include those described herein in any pharmaceutically acceptable form thereof, including isomers, salts, solvates, and polymorphs, as well as racemic mixtures and prodrugs thereof.

According to aspects described herein, the therapeutic regimens of the present disclosure are suitable for inhibiting the viral replication machinery of ssRNA viruses, and are also suitable for inhibiting cytopathic effects during ssRNA viral infection.

Small-molecule Hsps interact with a large number of client proteins that are crucial for many cellular processes. For example, Hsp90 interacts with over 200 polypeptides to regulate their activity and/or half-life. Hsp90, HspB1, and likely other small-molecule Hsps are global regulators of cellular systems. Hsp90 is a major factor in the replication of negative-strand viruses and is responsible for proper protein folding, intracellular disposal, protein stability against heat stress, and the proteolytic conversion of many essential regulators of cell growth and differentiation.

Brivudine (bromovinyl deoxyuridine or BVDU for short) interacts with two phenylalanine residues (Phe29 and Phe33) in the N-terminal domain of HspB1. The full chemical description of the drug is (E)-5-(2-bromovinyl)-2-deoxyuridine. Brivudine is a nucleoside analog that targets two viral enzymes: deoxythymidine kinase and polymerase. Brivudine can be incorporated into viral DNA and then blocks the action of DNA polymerase, thereby inhibiting viral replication. The present disclosure includes oral formulations of brivudine that can be used in methods for treating ssRNA viral infections. The active compound is the 5'-triphosphate of BVDU, which is formed by viral thymidine kinase and possibly by nucleoside diphosphate kinase upon subsequent phosphorylation. Brivudine can bind to the heat stress protein HSPB1 in vitro and inhibit its interaction with its binding partners. Brivudine has properties against HSP27, HSP70 and HSP90.

A non-limiting example of a prodrug form of BVDU is represented by Formula I:

Brivudine is represented by Formula 2:

The solid oral dosage composition of the present disclosure includes (E)-5-(2-bromovinyl) 2'-deoxyuridine (BVDU), a salt thereof, or a protected or prodrug form of BVDU, and at least one conventional carrier, and may include at least one auxiliary material. BVDU may be present in an amount effective to produce a concentration in the blood of 0.02 μg/ml to 10.0 μg/ml.

According to aspects described herein, a composition is disclosed comprising brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or BVDU in protected or prodrug form, wherein brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or BVDU in protected or prodrug form is present in an amount that, when administered to a patient suffering from a filovirus-mediated disease, is effective in treating the patient. In one embodiment, the virus is Ebola virus or Marburg virus.

According to aspects described herein, a method for treating a patient suffering from a filovirus-mediated disease comprises administering to the patient a composition comprising brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or BVDU in a protected or prodrug form in an amount effective to treat the patient. In one embodiment, the virus is Ebola virus or Marburg virus.

Upamostat ("WX-671" or "Mesupron") inhibits the urokinase-type plasminogen activator (uPA) system. Upamostat is a serine protease inhibitor. After oral administration, the serine protease inhibitor WX-671 is converted to the active Nα-(2,4,6-triisopropylphenylsulfonyl)-3-amidino-(L)-phenylalanine-4-ethoxycarbonylpiperazine ("WX-UK1"), which inhibits several serine proteases, particularly uPA. The serine protease inhibitor upamostat can potentially inhibit viral RNA replication. Oral formulations of the present invention comprising upamostat can be used in methods for treating ssRNA viral infections. The full chemical description of the drug is (S)-ethyl 4-(3-(3-(N-hydroxycarbamimidoyl)phenyl)-2-(2,4,6-triisopropylphenylsulfonylamino)propionyl)piperazine-1-carboxylate. In one embodiment of the present disclosure, upamostat is orally administered at a dose of about 0.5 to about 1.1 mg/kg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 400 mg. In one embodiment, upamostat is orally administered at a daily dose of about 150 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 250 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 300 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 350 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 400 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 450 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 500 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 500 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 450 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 350 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 300 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 250 mg. In one embodiment, upamostat is orally administered at a daily dose of about 500 mg to 1000 mg. In one embodiment, upamostat is orally administered at a daily dose of about 750 mg to 1000 mg. In one embodiment, upamostat is orally administered at a daily dose of about 500 mg to 750 mg.

Upamostat is represented by Formula 3:

In one embodiment of the present disclosure, upamostat is orally administered at a dose of about 0.5 to about 1.1 mg/kg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 400 mg. In one embodiment, upamostat is orally administered at a daily dose of about 150 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 250 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 300 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 350 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 400 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 450 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 500 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 550 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 500 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 450 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 350 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 300 mg. In one embodiment, upamostat is orally administered at a daily dose of about 200 mg to 250 mg. In one embodiment, upamostat is orally administered at a daily dose of about 500 mg to 1000 mg. In one embodiment, upamostat is orally administered at a daily dose of about 750 mg to 1000 mg. In one embodiment, upamostat is orally administered at a daily dose of about 500 mg to 750 mg.

Clofazimine is a riminophenazine compound that accumulates to very high concentrations in tissues. Clofazimine may inhibit bacterial growth by inducing host cell apoptosis. Treatment with clofazimine may result in highly condensed chromatin in the nucleus, indicating that macrophages are undergoing apoptosis. Macrophages are responsible for a range of biochemical products with potent immunomodulatory activities. In addition, clofazimine can stimulate the activity of various reticuloendothelial phagocytes. Cells, primarily monocytes and macrophages, accumulate in the lymph nodes and spleen, connective tissue and liver, lungs and central nervous system (microglia). The mononuclear phagocyte system is an integral part of humoral and cell-mediated immunity and plays an important role in the fight against microorganisms, including mycobacteria, fungi and viruses.

According to aspects described herein, a composition comprising clofazimine is disclosed, wherein the clofazimine is present in an amount that, when administered to a patient suffering from a filovirus-mediated disease, is effective in treating the patient. In one embodiment, the filovirus is Ebola virus or Marburg virus. According to aspects described herein, a method for treating a patient suffering from a filovirus-mediated disease comprises administering to the patient a composition comprising clofazimine in an amount effective in treating the patient. In one embodiment, the filovirus is Ebola virus or Marburg virus.

In one embodiment, the daily dose of clofazimine is from about 15 mg/day to about 120 mg/day until recovery. In one embodiment, the maximum dose of clofazimine is 80-120 mg/day until recovery. In one embodiment, clofazimine is administered orally twice daily as a 50 mg tablet. In one embodiment, clofazimine is administered orally once daily as a 100 mg tablet. In one embodiment, clofazimine is administered up to six times daily as a component of a solid oral dosage form comprising 10 mg to 16 mg of clofazimine per dosage form. In one embodiment, the method for treating an ssRNA viral infection (e.g., Ebola virus) comprises concomitantly administering: i) up to 120 mg/day of an oral clofazimine dosage form, ii) up to 450 mg/day of an oral rifabutin dosage form, iii) up to 1000 mg/day of clarithromycin as an intravenous infusion or a solid oral dosage form, and iv) up to 600 mg/day of an oral dosage form comprising brivudine, an active metabolite of brivudine, a salt thereof, or a protected or prodrug form of BVDU, wherein the administration continues for a period of time until the subject is free of and clears the ssRNA viral infection, such as Ebola virus. Clofazimine is represented by Formula 4:

Infection with ssRNA viruses, such as Ebola virus, induces massive lymphocyte apoptosis, which is thought to prevent the development of a functional adaptive immune response. Rifabutin may be effective in inhibiting the cytopathic effects of ssRNA viral infection, such as Ebola virus, which destroys T lymphocytes. Rifabutin interferes with the RNA replication machinery by inhibiting RNA synthesis (transcription), thereby initiating a new round of DNA replication.

According to aspects described herein, a composition comprising rifabutin is disclosed, wherein the rifabutin is present in an amount effective to treat the patient when administered to a patient suffering from a filovirus-mediated disease. In one embodiment, the filovirus is Ebola virus or Marburg virus. According to aspects described herein, a method for treating a patient suffering from a filovirus-mediated disease comprises administering to the patient a composition comprising rifabutin in an amount effective to treat the patient. In one embodiment, the filovirus is Ebola virus or Marburg virus.

In one embodiment, the daily dose of rifabutin is from about 80 mg/day to about 480 mg/day. In one embodiment, the maximum dose of rifabutin is 480 mg/day until recovery. In one embodiment, rifabutin is administered orally twice daily as a 150 mg tablet. In one embodiment, rifabutin is administered orally once daily as a 300 mg tablet. In one embodiment, rifabutin is administered up to 6 times daily as a component of a solid oral dosage form comprising 45 mg to 60 mg of rifabutin per dosage form. In one embodiment, the method for treating an ssRNA viral infection (e.g., Ebola virus) comprises concomitantly administering: i) up to 450 mg/day of an oral rifabutin dosage form, ii) up to 120 mg/day of an oral clofazimine dosage form, iii) up to 1000 mg/day of clarithromycin as an intravenous infusion or solid oral dosage form, and iv) up to 600 mg/day of an oral dosage form comprising brivudine, an active metabolite of brivudine, a salt thereof, or a protected or prodrug form of BVDU. Rifabutin is represented by Formula 5:

According to aspects described herein, a composition comprising clarithromycin is disclosed, wherein the clarithromycin is present in an amount effective to treat the patient when administered to a patient suffering from a filovirus-mediated disease. In one embodiment, the filovirus is Ebola virus or Marburg virus. According to aspects described herein, a method for treating a patient suffering from a filovirus-mediated disease comprises administering to the patient a composition comprising clarithromycin in an amount effective to treat the patient. In one embodiment, the filovirus is Ebola virus or Marburg virus.

In one embodiment, the daily dose of clarithromycin is from about 180 mg/day to about 1000 mg/day, until recovery. In one embodiment, the maximum dose of clarithromycin is 980-1000 mg/day, until recovery. In one embodiment, two doses of 500 mg clarithromycin are administered as an intravenous (IV) infusion using a solution concentration of about 2 mg/ml. 1 gram of clarithromycin is administered daily as an intravenous (IV) infusion for a period of two to five days. In one embodiment, 1 gram of clarithromycin is administered daily as an intravenous (IV) infusion for a period of three days. In one embodiment, clarithromycin is administered orally twice a day as a 500 mg tablet. In one embodiment, clarithromycin is administered as a component of a solid oral dosage form, which contains 95 mg to 125 mg of clarithromycin per dosage form. The solid dosage form can be administered up to twelve times a day. In one embodiment, the method for treating an ssRNA viral infection (e.g., Ebola virus) comprises intravenously administering 1 gram of clarithromycin per day for a first period of time, e.g., 2 to 5 days, followed by oral administration of 1 gram of clarithromycin per day for a second period of time, e.g., from the end of the first period of time until the subject is free of and clears the ssRNA viral infection, such as Ebola virus. In one embodiment, the method for treating an ssRNA viral infection (e.g., Ebola virus) comprises concomitantly administering: i) up to 1 gram/day of clarithromycin, ii) up to 600 mg/day of an oral dosage form comprising brivudine, an active metabolite of brivudine, a salt thereof, or BVDU in a protected or prodrug form, iii) up to 120 mg/day of an oral dosage form of clofazimine, and iv) up to 450 mg/day of an oral dosage form of rifabutin, wherein the administration is for a period of time until the subject is free of and clears the ssRNA viral infection, e.g., Ebola virus. Clarithromycin is represented by Formula 6:

Proinflammatory cytokines such as TNF-α, IL-1β, IL-6, IL-8, IL-12, and IFN-γ increase vascular permeability, promote vascular leakage and recruitment of neutrophils to sites of inflammation, and stimulate the production of acute phase proteins. In clinical settings, they can cause fever or hypothermia and peripheral shock. In animal models, co-administration of clarithromycin with clofazimine and/or rifabutin specifically and effectively reduces proinflammatory cytokines such as TNF-α and IL-6. During viral infection, macrophages secrete large amounts of proinflammatory cytokines such as TNF-α. The combination of the above drugs can effectively inhibit proinflammatory cytokines and reduce the permeability of vascular endothelial cells, which facilitates viral entry into endothelial cells.

Clarithromycin is an effective immunomodulator. Rifabutin can enhance the antibacterial and intercellular effects of clarithromycin. In addition, clarithromycin as an immunomodulator can have considerable efficacy in patients with sepsis (as sometimes seen in ssRNA viral infections such as Ebola virus infection). In one embodiment, clarithromycin is orally administered to a subject as a component of a solid oral dosage form. In one embodiment, clarithromycin is administered to a subject as an intravenous infusion. In one embodiment, clarithromycin can be administered to a subject as an intravenous infusion in combination with brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or an oral dosage form of BVDU in a protected or prodrug form. In one embodiment, clarithromycin can be administered to a subject as an intravenous infusion in combination with an oral dosage form of upamostat.

According to aspects described herein, compositions comprising rifabutin and clarithromycin are disclosed, wherein the rifabutin and clarithromycin are present in an amount effective to treat the patient when administered together to a patient suffering from a filovirus-mediated disease. In one embodiment, the filovirus is Ebola virus or Marburg virus. According to aspects described herein, methods for treating a patient suffering from a filovirus-mediated disease comprise administering to the patient a composition comprising rifabutin and clarithromycin in an amount effective to treat the patient. In one embodiment, the filovirus is Ebola virus or Marburg virus.

According to aspects described herein, a composition comprising clofazimine, rifabutin, and clarithromycin is disclosed, wherein the clofazimine, rifabutin, and clarithromycin are present in an amount effective to treat the patient when administered together to a patient suffering from a filovirus-mediated disease. In one embodiment, the filovirus is Ebola virus or Marburg virus. According to aspects described herein, a method for treating a patient suffering from a filovirus-mediated disease comprises administering to the patient a composition comprising clofazimine, rifabutin, and clarithromycin in an amount effective to treat the patient. In one embodiment, the filovirus is Ebola virus or Marburg virus.

In one embodiment, rifabutin, clarithromycin, and clofazimine are administered as a single solid oral dosage form. In one embodiment, a solid oral dosage form of the present disclosure comprises rifabutin, clarithromycin, clofazimine, and a pharmaceutically acceptable carrier, wherein the amount of clofazimine relative to the amount of clarithromycin is 5-18% w/w (e.g., 7-16%, 9-14%, 9-12%, 10-15%, or 0-11% w/w) and relative to the amount of rifabutin is 10-25% w/w (e.g., 12-25%, 12-23%, 15-25%, 15-23%, 18-25%, 18-23%, 20-25%, 20-23%, or 21-23%).

In one embodiment, the solid oral dosage form of the present disclosure comprises rifabutin, clarithromycin, and clofazimine in a ratio of 8-10:18-20:1-2.5 w/w/w (e.g., a ratio of 8.5-9.5:18.5-19.5:1.5-2.5 w/w/w or a ratio of 9:19:2, wherein each variable is freely varied by ±0.5 or 0.25). In one embodiment, the solid oral dosage form of the present disclosure comprises rifabutin, clarithromycin, and clofazimine in a ratio of about 9:19:2 w/w/w, wherein each variable is freely varied by ±2, 1, 0.5, or 0.25 (e.g., 9±0.5:19±5:2±0.5). For example, in one embodiment, a solid oral dosage form of the present disclosure comprises 90 mg rifabutin (±30, 20, 10, 5, 2, or 1 mg), 190 mg clarithromycin (±60, 40, 20, 10, 5, 2, or 1 mg), and 20 mg clofazimine (±10, 7, 5, 2, or 1 mg). In one embodiment, a solid oral dosage form of the present disclosure comprises 45 mg rifabutin (±15, 10, 7, 5, 2, or 1 mg), 95 mg clarithromycin (±30, 20, 10, 5, 2, or 1 mg), and 10 mg clofazimine (±6, 5, 2, or 1 mg).

In an embodiment, the solid oral dosage form of the present disclosure further comprises an absorption enhancer that can improve the bioavailability of one or more active ingredients. The amount of the absorption enhancer can be between 300-700% w/w relative to the amount of clofazimine, including 400-600% or 450-550% or 475-525%. In certain embodiments, the absorption enhancer is polyethylene glycol (PEG), such as a polyethylene glycol having an average molecular weight between 200 and 20,000, including 1000-15000 or 5000-12000 or 7000-9000 or 7500-8500, such as PEG8000).

In an embodiment, the solid oral dosage form of the present disclosure further comprises one or more additional excipients, such as MCC-Tabulose Type 200, magnesium stearate, SLS-Emal10Pwd HD, polysorbate (e.g., polysorbate 80), or a combination thereof, including all of these. In some cases, the present composition comprises polyethylene glycol and polysorbate (e.g., polysorbate 80), wherein the amount of polysorbate relative to the amount of clofazimine is 30-120% w/w (e.g., 50-100%, 50-85%, or 60-75%).

In one embodiment, the solid oral dosage form of the present disclosure further comprises one or more additional excipients, such as microcrystalline cellulose (MCC) SC 200), magnesium stearate, sodium lauryl sulfate (SLS) 10Pwd HD, polysorbate (e.g., polysorbate 80), or a combination thereof, including all of these. In some cases, the present composition comprises polyethylene glycol and polysorbate (e.g., polysorbate 80), wherein the amount of polysorbate relative to the amount of clofazimine is 30-120% w/w (e.g., 50-100%, 50-85%, or 60-75%).

In one embodiment, the solid oral dosage form of the present disclosure is provided in the form of a capsule or tablet containing the active substance in powder form. In one embodiment, the solid oral dosage form of the present disclosure is in the form of a capsule or tablet containing the active substance in microencapsulated form. In one embodiment, the solid oral dosage form of the present disclosure is in the form of a capsule or tablet containing the active substance in microparticle form.

In one embodiment, the solid oral dosage form of the present disclosure is available in the form of a tablet comprising at least one of rifabutin, clarithromycin, and clofazimine in powder form. In some cases, two or all of rifabutin, clarithromycin, and clofazimine are in powder form. In one embodiment, the solid oral dosage form of the present disclosure is in the form of a tablet or capsule comprising at least one of rifabutin, clarithromycin, and clofazimine in microencapsulated form. In some cases, two or all of rifabutin, clarithromycin, and clofazimine are in microencapsulated form. In one embodiment, the solid oral dosage form of the present disclosure is in the form of a tablet or capsule comprising at least one of rifabutin, clarithromycin, and clofazimine in powder form, with the remaining agents being in microcapsule form. In one embodiment, the solid oral dosage form of the present disclosure is in the form of a tablet or capsule comprising one or more of rifabutin, clarithromycin, and clofazimine in microsomal form. In one embodiment, the solid oral dosage form of the present disclosure is in the form of a tablet containing one or more of rifabutin, clarithromycin, and clofazimine in a capsule, or in the form of a capsule containing one or more of rifabutin, clarithromycin, and clofazimine in a tablet, or in the form of a capsule containing one or more of rifabutin, clarithromycin, and clofazimine in an outer capsule containing other agents, or any combination thereof. In one embodiment, the solid oral dosage form of the present disclosure comprises an inner capsule containing rifabutin in an outer capsule containing clarithromycin and clofazimine, wherein the clarithromycin and clofazimine may be in powdered, microcapsulated, or particulate form.

Surprisingly, the combination of multiple drugs disclosed herein, including but not limited to brivudine in combination with at least one of clofazimine, rifabutin, clarithromycin, or upamostat, can inhibit the replication mechanism of ssRNA viral infection, thereby hindering, inhibiting, or preventing viral infection.

Surprisingly, the combination of multiple drugs disclosed herein, including but not limited to brivudine in combination with at least one of clofazimine, rifabutin, clarithromycin, or upamostat, can inhibit the cytopathic effects of ssRNA viral infection, thereby hindering, inhibiting, or preventing degenerative changes or abnormalities.

According to aspects described herein, a composition comprising rifabutin, clarithromycin, and brivudine is disclosed, wherein the rifabutin, clarithromycin, and brivudine are present in amounts effective to treat a patient suffering from a filovirus-mediated disease when administered together. In one embodiment, the filovirus is Ebola virus or Marburg virus.

According to aspects described herein, a method for treating a patient suffering from a filovirus-mediated disease comprises administering to the patient a composition comprising rifabutin, clarithromycin, and brivudine in an amount effective to treat the patient. In one embodiment, the filovirus is Ebola virus or Marburg virus.

According to aspects described herein, a method for treating a subject having an ssRNA viral infection comprises administering in combination a therapeutically effective amount of brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU; a therapeutically effective amount of rifabutin; and a therapeutically effective amount of clarithromycin.

According to aspects described herein, a method for treating a subject having an ssRNA viral infection comprises the combination of: oral administration of a tablet containing a therapeutically effective amount of brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU; oral administration of a capsule or tablet containing rifabutin; and oral administration of a capsule or tablet containing clarithromycin, wherein rifabutin and clarithromycin can be present in the same capsule or tablet.

According to aspects described herein, methods for treating a subject with an ssRNA viral infection comprise the following combination: oral administration of a therapeutically effective amount of brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU for a suitable period of time; and administration of a therapeutically effective amount of at least one of clofazimine, rifabutin, or clarithromycin for a suitable period of time. In one embodiment, the therapeutically effective amount of BVDU is up to 600 mg/day for adults. In one embodiment, 600 mg is administered once daily as a single oral dosage form. In one embodiment, 600 mg is administered as a single 150 mg oral dosage form, taken four times daily. In one embodiment, the therapeutically effective amount of BVDU is up to 500 mg/day for adults. In one embodiment, 500 mg is administered once daily as a single oral dosage form. In one embodiment, 500 mg is administered as a single 125 mg oral dosage form, taken four times daily. In one embodiment, the therapeutically effective amount of clofazimine for adults is from about 50 mg to about 300 mg per day. In one embodiment, clofazimine is administered orally once or more daily as a solid dosage form. In one embodiment, the therapeutically effective amount of rifabutin for an adult is about 45 mg to about 480 mg per day. In one embodiment, rifabutin is administered orally once or more daily as a solid dosage form. In one embodiment, the therapeutically effective amount of clofazimine for an adult is about 50 mg to about 300 mg per day. In one embodiment, clofazimine is administered orally once or more daily as a solid dosage form.

According to aspects described herein, a method for treating a subject with an ssRNA viral infection comprises the following combination: oral administration of a therapeutically effective amount of brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU for a suitable period of time; oral administration of a therapeutically effective amount of a solid oral dosage form comprising at least one of clofazimine or rifabutin for a suitable period of time; and administration of a therapeutically effective amount of clarithromycin as an intravenous infusion for a suitable period of time. In one embodiment, for adults, the therapeutically effective amount of BVDU is up to 600 mg/day. In one embodiment, 600 mg is administered once daily as a single oral dosage form. In one embodiment, 600 mg is administered as a 150 mg single oral dosage form taken four times daily. In one embodiment, the therapeutically effective amount of BVDU is up to 500 mg/day for adults. In one embodiment, 500 mg is administered once daily as a single oral dosage form. In one embodiment, 500 mg is administered as a 125 mg single oral dosage form taken four times daily. In one embodiment, for adults, the therapeutically effective amount of clarithromycin is up to 1 gram per day. In one embodiment, two doses of 500 mg of clarithromycin are administered as an IV infusion using a solution concentration of about 2 mg/ml. 1 gram of clarithromycin per day can be administered as an IV infusion for a period of two to five days. In one embodiment, 1 gram of clarithromycin per day can be administered as an intravenous infusion for three days. In one embodiment, the therapeutically effective amount of rifabutin per day for adults is up to 480 mg. In one embodiment, rifabutin is administered orally as a tablet once or more daily. In one embodiment, rifabutin is administered as a component of a solid oral dosage form containing 45 mg to 60 mg of rifabutin per dosage form. A solid dosage form containing 45 mg to 60 mg of rifabutin can be administered up to twelve times per day. In one embodiment, the therapeutically effective amount of clofazimine per day for adults is about 50 mg to about 300 mg. In one embodiment, clofazimine is administered orally as a solid dosage form once or more daily. In one embodiment, clofazimine is administered as a component of a solid oral dosage form comprising 10 mg to 16 mg of clofazimine per dose.

According to aspects described herein, methods for treating a subject with an ssRNA viral infection comprise the following combination: oral administration of a therapeutically effective amount of brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU for a suitable period of time; and administration of a therapeutically effective amount of clarithromycin as an intravenous infusion for a suitable period of time. In one embodiment, the therapeutically effective amount of BVDU is up to 600 mg/day for adults. In one embodiment, 600 mg is administered once daily as a single oral dosage form. In one embodiment, 600 mg is administered as a 150 mg single oral dosage form, taken four times daily. In one embodiment, the therapeutically effective amount of BVDU is up to 500 mg/day for adults. In one embodiment, 500 mg is administered once daily as a single oral dosage form. In one embodiment, 500 mg is administered as a 125 mg single oral dosage form, taken four times daily. In one embodiment, the therapeutically effective amount of clarithromycin for adults is up to 1 gram per day. Clarithromycin is administered as an intravenous (IV) infusion. In one embodiment, two doses of 500 mg clarithromycin are administered as IV infusions using a solution concentration of about 2 mg/ml. 1 gram of clarithromycin per day can be administered as an IV infusion for a period of two to five days. In one embodiment, 1 gram of clarithromycin per day can be administered as an intravenous infusion for three days.

According to aspects described herein, a method for treating a subject with an ssRNA viral infection comprises the following combination: oral administration of a therapeutically effective amount of brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or BVDU in a protected or prodrug form for a suitable period of time; oral administration of a therapeutically effective amount of a solid oral dosage form comprising clarithromycin for a suitable period of time; oral administration of a therapeutically effective amount of a solid oral dosage form comprising rifabutin for a suitable period of time; and oral administration of a therapeutically effective amount of a solid oral dosage form comprising clofazimine for a suitable period of time. In one embodiment, the therapeutically effective amount of BVDU for an adult is up to 600 mg/day. In one embodiment, 600 mg is administered once daily as a single oral dosage form. In one embodiment, 600 mg is administered as a 150 mg single oral dosage form taken four times daily. In one embodiment, the therapeutically effective amount of BVDU for an adult is up to 500 mg/day. In one embodiment, 500 mg is administered once daily as a single oral dosage form. In one embodiment, 500 mg is administered as a 125 mg single oral dosage form taken four times daily. In one embodiment, for adults, the therapeutically effective amount of clarithromycin is up to 1 gram per day. In one embodiment, clarithromycin is administered orally twice daily as a 500 mg tablet. In one embodiment, for adults, the therapeutically effective amount of rifabutin is up to 480 mg per day. In one embodiment, rifabutin is administered orally once or multiple times per day as a tablet. In one embodiment, rifabutin is administered as a component of a solid oral dosage form containing 45 mg to 60 mg of rifabutin per dosage form. A solid dosage form containing 45 mg to 60 mg of rifabutin can be administered up to twelve times per day. In one embodiment, for adults, the therapeutically effective amount of clofazimine is about 50 mg to about 300 mg per day. In one embodiment, clofazimine is administered orally once or multiple times per day as a solid dosage form. In one embodiment, clofazimine is administered as a component of a solid oral dosage form containing 10 mg to 16 mg of clofazimine per dose.

In one embodiment, clofazimine and brivudine are orally administered as a solid dosage form once or more per day. In one embodiment, clofazimine, rifabutin, and brivudine are orally administered as a solid dosage form once or more per day. In one embodiment, rifabutin and brivudine are orally administered as a solid dosage form once or more per day. In one embodiment, clarithromycin, rifabutin, and brivudine are orally administered as a solid dosage form once or more per day. In one embodiment, clarithromycin, clofazimine, and brivudine are orally administered as a solid dosage form once or more per day. In one embodiment, upamostat, clofazimine, and brivudine are orally administered as a solid dosage form once or more per day. In one embodiment, upamostat, rifabutin, clofazimine, and brivudine are orally administered as a solid dosage form once or more per day. In one embodiment, upamostat, rifabutin, clarithromycin, and brivudine are orally administered as a solid dosage form once or more per day. In one embodiment, upamostat, brivudine, clarithromycin, and clofazimine are administered orally one or more times daily as a solid dosage form. In one embodiment, upamostat, brivudine, clarithromycin, rifabutin, and clofazimine are administered orally one or more times daily as a solid dosage form.

In one embodiment, rifabutin, clarithromycin, and clofazimine are administered in a single solid oral dosage form. In some cases, rifabutin, clarithromycin, and clofazimine are co-administered once daily for a first period of treatment (e.g., 1-3 weeks, including 1 week, 2 weeks, or 3 weeks) in an amount as follows: (i) 80-100 mg rifabutin (e.g., 85-95 mg or 90 mg ± 1.5 mg), (ii) 180-200 mg clarithromycin (e.g., 185-195 mg or 190 mg ± 2 mg), and (iii) 15-25 mg clofazimine (e.g., 17-23 mg or 20 ± 1 mg). The method may further comprise the step of linearly increasing the amounts of rifabutin, clarithromycin, and clofazimine while maintaining a ratio of 8-10:18-20:1-2.5 w/w/w (e.g., a ratio of 8.5-9.5:18.5-19.5:1.5-2.5 w/w/w or a ratio of 9:19:2, wherein each variable can freely vary by ±0.5 or 0.25), for a second period of treatment (e.g., 4-10 weeks). In one embodiment, the linearly increasing amounts of rifabutin, clarithromycin, and clofazimine during the second period of treatment do not exceed (i) 420-480 mg rifabutin (e.g., 440-460 mg or 450 mg), 920-980 mg clarithromycin (e.g., 940-960 mg or 950 mg), and (iii) 80-120 mg clofazimine (e.g., 90-110 mg or 100 mg). In some cases, the linear increasing amounts of rifabutin, clarithromycin, and clofazimine include: a) (i) 160-200 mg rifabutin (e.g., 170-190 mg or 180 mg ± 2 mg), (ii) 360-400 mg clarithromycin (e.g., 370-390 mg or 380 mg ± 2 mg), and (iii) 30-50 mg clofazimine (e.g., 35-45 mg or 40 mg ± 1 mg), once daily for two weeks; b) (i) 250-290 mg rifabutin (e.g., 260-280 mg or 270 mg ± 2 mg), (ii) 550-590 mg clarithromycin (e.g., 560-580 mg or 570 ± 2 mg), and (iii) 50-70 mg clofazimine (e.g., 55-65 mg or 60 mg ± 1.5 mg) , once daily for two weeks; c) (i) 340-380 mg rifabutin (e.g., 350-370 mg or 360 mg ± 2 mg), (ii) 740-780 mg clarithromycin (e.g., 750-770 mg or 760 mg ± 2 mg), and (iii) 60-100 mg clofazimine (e.g., 70-90 mg or 80 mg ± 1.5 mg), once daily for two weeks; and d) (i) 420-480 mg rifabutin (e.g., 440-460 mg or 450 mg ± 2 mg), (ii) 920-980 mg clarithromycin (e.g., 940-960 mg or 950 mg ± 2 mg), and (iii) 80-120 mg clofazimine (e.g., 90-110 mg or 100 mg ± 1.5 mg), once daily for one week. In certain embodiments, the method further comprises, after step d) above, co-administering (i) 420-480 mg rifabutin (e.g., 440-460 mg or 450 mg ± 2 mg), (ii) 920-980 mg clarithromycin (e.g., 940-960 mg or 950 mg ± 2 mg), and (iii) 80-120 mg clofazimine (e.g., 90-110 mg or 100 mg ± 1.5 mg) once daily for a third period of treatment. In some embodiments, the third period of treatment is 1, 2, 4, 6, 8, 12 weeks; 3, 6, or 12 months or longer. In one embodiment, the third period of treatment continues until symptoms associated with the ssRNA viral infection are alleviated or the ssRNA viral disease is controlled as indicated by laboratory diagnosis.

In one embodiment, a single solid oral dosage form comprises rifabutin; clarithromycin; clofazimine; an absorption enhancer; and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition is a solid oral dosage form, wherein the amount of the absorption enhancer relative to the amount of clofazimine is 300% to 700% w/w, and wherein the amount of clofazimine relative to the amount of clarithromycin is 10-15% w/w and relative to the amount of rifabutin is 20-25% w/w.

In one embodiment, a single solid oral dosage form comprises rifabutin; clarithromycin; clofazimine; polyethylene glycol; and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition is a solid oral dosage form, wherein the polyethylene glycol (i) has an average molecular weight of 1,000-15,000 Daltons, and (ii) is 300% to 700% w/w relative to the amount of rifabutin, and wherein the amount of clofazimine is 10-15% w/w relative to the amount of clarithromycin and 20-25% w/w relative to the amount of rifabutin.

Small molecule heat shock proteins (small molecule Hsps) are stress-induced molecular chaperones that act as holdases for polypeptides that have lost their folding under stress conditions or have therefore mutated in their coding sequences. Therefore, cytoprotection is provided to cells to prevent the harmful effects of damaged protein-mediated damage. These chaperones are also highly expressed in response to protein conformation and inflammatory diseases and cancer pathology. Through the specific and reversible modification of their phosphate oligomerization, small molecule Hsps can be accompanied by appropriate receptor proteins to provide cells with resistance to different types of damage or pathological conditions. By helping cells better cope with their pathological conditions, their expression can be beneficial, for example in diseases characterized by pathological cell degeneration, or when they are necessary for tumor cell survival, their expression is harmful. In addition, small molecule Hsps are actively released by cells and can serve as immunogenic molecules with a dual effect that depends on pathology. Five Hsps families are induced by stress: the 70 kDa (HspA-Hsp70) family, the 20-30 kDa (HspB-small Hsps, sHsps) family, the 90 kDa (HspC-Hsp90) family, the 60 kDa (HspD-Hsp60) family, and the HspH (large Hsps) family.

The aryl adamantane compounds of the present invention have been shown to selectively inhibit 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 by cells expressing cellular proteins that serve as receptors for Ebola and Marburg viruses. According to aspects described herein, a composition comprising an aryl adamantane compound is disclosed, wherein the aryl adamantane compound is present in an amount effective to treat the patient when administered to a patient suffering from a filovirus-mediated disease. In one embodiment, the filovirus is Ebola or Marburg. According to aspects described herein, a method for treating a patient suffering from a filovirus-mediated disease comprises administering to the patient a composition comprising an aryl adamantane compound in an amount effective to treat the patient. In one embodiment, the filovirus is Ebola or Marburg.

The symbol "-" generally represents a bond between two atoms in a chain. Thus, _CH3 -O- _CH2 -CH( _R1 ) _-CH3 represents a 2-substituted-1-methoxypropane compound. Additionally, the symbol "-" indicates the point of attachment of a substituent to a compound. Thus, for example, aryl( _C1 - _C6 )alkyl- represents an alkylaryl group attached to the compound at the alkyl portion, such as benzyl.

When multiple substituents are indicated as being attached to a structure, it is understood that the substituents may be the same or different. Thus, for example, " _Rm optionally substituted with 1, 2, or 3 _Rq groups" means that _Rm is substituted with 1, 2, or 3 _Rq groups, wherein the _Rq groups may be the same or different.

The term "optionally substituted" is used interchangeably with the phrase "substituted or unsubstituted." Unless otherwise specified, an optionally substituted group can have a substituent at each substitutable position of the group, and each substituent is not constrained by another substituent.

As used herein, the term "halogen" or "halo" refers to fluorine, chlorine, bromine, or iodine.

The term "heteroatom" refers to nitrogen, oxygen or sulfur, and includes any oxidized forms of nitrogen and sulfur, as well as quaternized forms of any basic nitrogen. The term "nitrogen" also includes substitutable nitrogen in heterocyclic rings. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from nitrogen, oxygen or sulfur, nitrogen can be N (such as 3,4-dihydro-2H-pyrrolyl), NH (such as pyrrolidinyl) or NR+ (such as in N-substituted pyrrolidinyl).

The term "alkyl" used alone or as part of a larger moiety refers to a saturated aliphatic hydrocarbon including straight chain, branched chain or cyclic (also referred to as "cycloalkyl") groups. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, iso-, sec- and tert-butyl, pentyl, hexyl, heptyl, 3-ethylbutyl, etc. Preferably, the alkyl group has 1 to 20 carbon atoms (whenever a numerical range is specified herein, such as "1-20", this means that the group in this case can contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms). More preferred are medium-sized alkyl groups with 1 to 10 carbon atoms. Most preferred are lower alkyl groups with 1 to 4 carbon atoms. Cycloalkyl groups can be monocyclic or polycyclic fused systems. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and adamantyl. Alkyl or cycloalkyl groups can be unsubstituted or substituted with 1, 2, 3 or more substituents. Examples of such substituents include, but are not limited to, halogen, hydroxy, amino, alkoxy, alkylamino, dialkylamino, cycloalkyl, aryl, aryloxy, arylalkoxy, heterocyclyl, and (heterocyclyl)oxy. Examples include fluoromethyl, hydroxyethyl, 2,3-dihydroxyethyl, (2- or 3-furyl)methyl, cyclopropylmethyl, benzyloxyethyl, (3-pyridyl)methyl, (2-thienyl)ethyl, hydroxypropyl, aminocyclohexyl, 2-dimethylaminobutyl, methoxymethyl, N-pyridylethyl, and diethylaminoethyl.

The term "cycloalkylalkyl" as used herein alone or as part of a larger moiety, refers to a _C3 - _C10 cycloalkyl group attached to the parent molecular moiety through an alkyl group, as defined above. Examples of cycloalkylalkyl groups include cyclopropylmethyl and cyclopentylethyl.

The term "alkenyl" as used herein alone or as part of a larger group refers to an aliphatic hydrocarbon having at least one carbon-carbon double bond, including straight, branched 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-sized alkenyl group having 2 to 10 carbon atoms. Most preferably, it is a lower alkenyl group having 2 to 6 carbon atoms. The alkenyl group can be unsubstituted or substituted with 1, 2, 3 or more substituents. Examples of these substituents include, but are not limited to, halogen, hydroxyl, amino, alkoxy, alkylamino, dialkylamino, cycloalkyl, aryl, aryloxy, arylalkoxy, heterocyclic and (heterocyclic)oxy. Depending on the position of the double bond and the substituent (if any), the geometry of the double bond can be opposite (E) or the same (Z), cis or trans. Examples of alkenyl include ethenyl, propenyl, cis-2-butenyl, trans-2-butenyl and 2-hydroxy-2-propenyl.

The term "alkynyl" as used herein alone or as part of a larger group refers to an aliphatic hydrocarbon having at least one carbon-carbon triple bond, including straight, branched or cyclic groups having at least one carbon-carbon triple bond. Alkynyl preferably has 2 to 20 carbon atoms. More preferably, it is a medium-sized alkynyl having 2 to 10 carbon atoms. Most preferably, it is a lower alkynyl having 2 to 6 carbon atoms. Alkynyl can be unsubstituted or substituted with 1, 2, 3 or more substituents. Examples of these substituents include, but are not limited to, halogen, hydroxyl, amino, alkoxy, alkylamino, dialkylamino, cycloalkyl, aryl, aryloxy, arylalkoxy, heterocyclic and (heterocyclic)oxy. Examples of alkynyl include ethynyl, propynyl, 2-butynyl and 2-hydroxy-3-butyl.

The term "alkoxy" as used herein alone or as part of a larger moiety refers to an alkyl group having a specified number of carbon atoms attached to the parent molecular moiety via an oxygen bridge. Examples of alkoxy groups include, for example, methoxy, ethoxy, propoxy, and isopropoxy. Alkoxy groups may be further substituted with one or more halogen atoms, such as fluorine, chlorine, or bromine, to provide a "haloalkoxy group." Examples of these groups include fluoromethoxy, chloromethoxy, trifluoromethoxy, and fluoroethoxy.

The term "aryl" as used herein alone or as part of a larger group refers to an aromatic hydrocarbon ring system containing at least one aromatic ring. The aromatic rings may optionally be fused or otherwise connected to other aromatic hydrocarbon rings or non-aromatic hydrocarbon rings. In addition, the aryl group may be substituted or unsubstituted by various groups such as hydrogen, halogen, hydroxyl, alkyl, haloalkyl, alkoxy, nitro, cyano, alkylamine, carboxyl or alkoxycarbonyl. Examples of aryl groups include, for example, phenyl, naphthyl, 1,2,3,4-tetralin, benzodioxole and biphenyl. Preferred examples of unsubstituted aryl groups include phenyl and biphenyl. Preferred aryl substituents include hydrogen, halogen, alkyl, haloalkyl, hydroxyl and alkoxy.

The term "heteroalkyl" as used herein alone or as part of a larger moiety refers to an alkyl group as defined herein in which one or more heteroatoms replace a carbon atom in the moiety. Such heteroalkyl groups are interchangeably used with the terms ether, thioether, amine, etc.

The term "heterocyclyl" as used herein, alone or as part of a larger group, refers to saturated, partially unsaturated, and unsaturated heteroatom-containing cyclic groups, wherein the heteroatom can be selected from nitrogen, sulfur, and oxygen. The heterocyclyl group can be unsubstituted or substituted at one or more atoms in the ring system. The heterocycle can contain one or more oxo groups.

The term "heterocycloalkyl" as used herein, alone or as part of a larger group, refers to a non-aromatic ring system containing at least one heteroatom selected from nitrogen, oxygen, and sulfur. The heterocycloalkyl ring may optionally be fused or otherwise connected to other heterocycloalkyl rings and/or non-aromatic hydrocarbon rings. Preferred heterocycloalkyl groups have 3 to 7 members. Examples of heterocycloalkyl groups include, for example, piperazine, morpholine, piperidine, tetrahydrofuran, pyrrolidine, and pyrazole. Preferred monocyclic heterocycloalkyl groups include piperidinyl, piperazinyl, morpholinyl, pyrrolidinyl, thiomorpholinyl, thiazolidinyl, 1,3-dioxolanyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydrothiopyranyl, and the like. Heterocycloalkyl groups may also be partially unsaturated. Examples of these groups include dihydrothienyl, dihydropyranyl, dihydrofuranyl, and dihydrothiazolyl.

The term "heteroaryl" as used herein alone or as part of a larger group refers to an aromatic ring system containing at least one heteroatom selected from nitrogen, oxygen and sulfur. The heteroaryl ring can be fused to or otherwise connected to one or more heteroaryl rings, aromatic or non-aromatic hydrocarbon rings or heterocycloalkyl rings. In addition, the heteroaryl group can be unsubstituted or substituted at one or more atoms of the ring system, or can 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, pyrimidinyl, imidazolyl, benzimidazolyl, furyl, benzofuranyl, thiazolyl, benzothiazolyl, isoxazolyl, oxadiazolyl, isothiazolyl, benzisothiazolyl, triazolyl, tetrazolyl, pyrrolyl, indolyl, pyrazolyl, benzopyrazolyl, purinyl, benzoxazolyl and carbazolyl.

The term "acyl" refers to a H-C(O)- or alkyl-C(O)- group, wherein the alkyl, straight chain, branched chain or cyclic group is as described above. Exemplary acyl groups include formyl, acetyl, propionyl, 2-methylpropionyl, butyryl and hexanoyl.

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.

The term "solvate" refers to a physical association of a compound of the invention with one or more solvent molecules. This physical association involves varying degrees 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 into the crystal lattice of a crystalline solid. "Solvate" includes both solution-phase and isolatable solvates. Exemplary solvates include ethanolates, methanolates, and the like. "Hydrate" is a solvate in which the solvent molecule is _H2O .

Examples of the aryl adamantane compounds of the present invention are generally represented by Formula 7, as shown below:

and pharmaceutically acceptable salts thereof, wherein

L is a bond or -C(R ₃ ,R ₄ )-;

_Where )S(O) ₂ -;

R ₁ is 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, -NO ₂ , -NH ₂ , -CO ₂ (alkyl), -OC(O)alkyl, carbamoyl, mono- or dialkylaminocarbamoyl, mono- or dialkylcarbamoyl, mono- or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, or mono- or dialkylthiocarbamoyl; R ₂ is 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, -NO ₂ , -NH ₂ , -CO ₂ (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, -C(O)-NH-aryl, -alkenyl-heteroaryl, -C(O)-heteroaryl, or -alkenyl-heteroaryl-aryl;

_R3 is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl, halo, 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 portions of each of the aforementioned R ₁ , R ₂ and R ₃ groups are optionally substituted with up to 5 groups which are independently (C ₁ -C ₆ ) alkyl, halogen, haloalkyl, -OC(O)(C ₁ -C ₆ alkyl), -C(O)O(C ₁ -C ₆ alkyl), -CONR'R", -OC(O)NR'R", -NR'C(O)R", -CF ₃ , -OCF ₃ , -OH, C ₁ -C ₆ alkoxy, hydroxyalkyl, -CN, -CO ₂ H, -SH, -S-alkyl, -SOR'R", -SO ₂ R', -NO ₂ or NR'R", wherein R' and R" are independently H or (C ₁ -C ₆ ) alkyl, and wherein each alkyl portion of the substituent is optionally further substituted with 1 , 2 or 3 groups independently selected from halogen, CN, OH and NH ₂ ; and

_R4 and _R5 are independently H or alkyl, provided that when _R3 and _R4 are on the same carbon and _R3 is oxo, then _R4 is absent.

The aryl adamantane compounds of formula 7 include compounds of formula I-1:

and pharmaceutically acceptable salts thereof, wherein:

R ₁ is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl, halo, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, -COOH, -OH, -SH, -S-alkyl, -CN, -NO ₂ , -NH ₂ , -CO ₂ (alkyl), -OC(O)alkyl, carbamoyl, mono- or dialkylaminocarbamoyl, mono- or dialkylcarbamoyl, mono- or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, or mono- or dialkylthiocarbamoyl; and

_R2 is 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-cycloalkyl or -alkenyl-heteroaryl-aryl,

wherein the alkyl and ring portions of each of the above R ₁ and R ₂ groups are optionally substituted with up to 5 groups independently selected from (C ₁ -C ₆ )alkyl, halogen, haloalkyl, -OC(O)(C ₁ -C ₆ alkyl), -C(O)O(C ₁ -C ₆ alkyl), -CONR'R", -OC(O)NR'R", -NR'C(O)R", -CF ₃ , -OCF ₃ , -OH, C ₁ -C ₆ alkoxy, hydroxyalkyl, -CN, -CO ₂ H, -SH, -S-alkyl, -SOR'R", -SO ₂ R', -NO ₂ or NR'R", wherein R' and R" are independently H or (C ₁ -C ₆ )alkyl, and wherein each alkyl portion of the substituent is optionally further substituted with 1, 2 or 3 groups independently selected from halogen, CN, OH, NH ₂ .

Aryladamantane compounds of Formula 7 include those of Formula II:

and pharmaceutically acceptable salts thereof, wherein:

Y is -C(R ₄ ,R ₅ )-, -N(R ₄ )-, -O- or -C(O)-;

R ₁ is 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, -NO ₂ , -NH ₂ , -CO ₂ (alkyl), -OC(O)alkyl, carbamoyl, mono- or dialkylaminocarbamoyl, mono- or dialkylcarbamoyl, mono- or dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl, or mono- or dialkylthiocarbamoyl; R ₂ is 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, -NO ₂ , -NH ₂ , -CO ₂ (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, -C(O)-NH-aryl, -alkenyl-heteroaryl, -C(O)-heteroaryl, or -alkenyl-heteroaryl-aryl;

_R3 is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl, heteroalkyl, aryl, alkaryl, alkenylaryl, heterocyclyl, heteroaryl, alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl, halo, 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 portions of each of the above R ₁ , R ₂ and R ₃ groups are optionally substituted with up to 5 groups which are independently (C ₁ -C ₆ ) alkyl, halogen, haloalkyl, -OC(O)(C ₁ -C ₆ alkyl), -C(O)O(C ₁ -C ₆ alkyl), -CONR'R", -OC(O)NR'R", -NR'C(O)R", -CF ₃ , -OCF ₃ , -OH, C ₁ -C ₆ alkoxy, hydroxyalkyl, -CN, -CO ₂ H, -SH, -S-alkyl, -SOR'R", -SO ₂ R', -NO ₂ or NR'R", wherein R' and R" are independently H or (C ₁ -C ₆ ) alkyl, and wherein each alkyl portion of the substituent is optionally further substituted with 1 , 2 or 3 groups independently selected from halogen, CN, OH, NH ₂ ; and

_R4 and _R5 are independently H or alkyl.

Compounds of Formula II include those wherein:

Y is -C(R ₄ ,R ₅ )- or -N(R ₄ )-;

wherein the alkyl and ring portions of each of the above R ₁ and R ₂ groups are optionally substituted with up to 5 groups which are independently (C ₁ -C ₆ ) alkyl, halogen, haloalkyl, -OC(O)(C ₁ -C ₆ alkyl), -C(O)O(C ₁ -C ₆ alkyl), -CONR ₄ R ₅ , -OC(O)NR ₄ R ₅ , -NR ₄ C(O)R ₅ , -CF ₃ , -OCF ₃ , -OH, C ₁ -C ₆ alkoxy, hydroxyalkyl, -CN, -CO ₂ H, -SH, -S-alkyl, -SOR ₄ R ₅ , -SO ₂ R ₄ R ₅ , -NO ₂ or NR ₄ R ₅ , and wherein each alkyl portion of the substituent is optionally further substituted with 1 , 2 or 3 groups independently selected from halogen, CN, OH, NH ₂ ;

_R3 is H, alkyl or oxo (=O); and

R ₄ and R ₅ are independently H or (C ₁ -C ₆ )alkyl.

Representative compounds of Formula II include:

Representative compounds of formula I-1 include:

A particularly preferred aromatic adamantane compound of the present invention is shown below and is referred to as ABC294640 [3-(4-chlorophenyl)-adamantane-1-carboxylic acid (pyridin-4-ylmethyl)amide]:

In one embodiment, the aromatic adamantane compound of the present invention is selected from the compound of formula 8:

and pharmaceutically acceptable salts thereof, wherein

○ _R1 is H, Cl or F;

○ _R2 is H or alkyl;

○m is 0, 1 or 2;

○n is 1, 2, 3, 4 or 5;

each R ₃ is independently H, -C(O)alkyl, -C(O)CH ₂ CH ₂ C(O)OH, R ₄ , -C(O)NR ₅ R ₆ , -P(O)(OR ₇ ) ₂ or glucosyl, provided that at least one R ₃ is not H,

○Among them

R4 is a natural or unnatural amino acid linked as an ester via the carboxyl moiety,

● _R5 is H or alkyl,

● _R6 is H or alkyl, and

Each R ₇ is independently H or alkyl.

In certain embodiments of the compound of formula (I) as described above,

The moiety is a catechol having at least one catechol-OH group substituted. For example, in one embodiment, the

The part has the following structure

In a particularly preferred embodiment of the compound of formula (I) as described above,

The part has the following structure

In a particularly preferred embodiment of the invention, the compound of formula (I) has R ₁ ═Cl, R ₂ ═H, m=2, n=2, and each R ₃ ═—C(O)alkyl, in particular —C(O)CH ₃ .

For example, compounds of the present invention include:

2-Acetoxy-5-(2-{[3-(4-chlorophenyl)-adamantane-1-carbonyl]-amino}ethyl)phenyl acetate;

2-Propionyloxy-5-(2-{[3-(4-chlorophenyl)-adamantane-1-carbonyl]-amino}ethyl)phenyl propionate;

2-Butyryloxy-5-(2-{[3-(4-chlorophenyl)-adamantane-1-carbonyl]-amino}ethyl)phenyl butyrate;

5-(2-{[3-(4-chlorophenyl)adamantane-1-carbonyl]amino}ethyl)-2-hydroxyphenyl isobutyrate; and

2-Amino-3-methyl-butyric acid 5-(2-{[3-(4-chlorophenyl)adamantane-1-carbonyl]amino}ethyl)-2-hydroxyphenyl ester.

A particularly preferred aromatic adamantane compound of the present invention is shown below and is referred to as ABC294735 [3-(4-chlorophenyl)adamantane-1-carboxylic acid [(3,4-dihydroxyphenyl)ethyl]amide]:

Solid dosage forms for oral administration can contain pharmaceutically acceptable adhesives, sweeteners, disintegrants, diluents, flavorings, coatings, preservatives, lubricants and/or time delay agents. Suitable adhesives include gum arabic, gelatin, corn starch, tragacanth gum, sodium alginate, carboxymethyl cellulose or polyethylene glycol (PEG). Suitable sweeteners include sucrose, lactose, glucose, aspartame or saccharin. Suitable disintegrants include corn starch, methylcellulose, polyvinyl pyrrolidone, xanthan gum, bentonite, alginic acid or agar. Suitable diluents include lactose, sorbitol, mannitol, glucose, kaolin, cellulose, calcium carbonate, calcium silicate or dicalcium phosphate. Suitable flavorings include peppermint oil, wintergreen oil, cherry, orange or raspberry flavoring. Suitable coating agents include polymers or copolymers of acrylic acid and/or methacrylic acid and/or its esters, wax, fatty alcohol, zein, shellac or gluten. Suitable preservatives include sodium benzoate, vitamin E, alpha-tocopherol, ascorbic acid, methylparaben, propylparaben or sodium bisulfite. Suitable lubricants include magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc. Suitable delay agents include glyceryl monostearate or glyceryl distearate.

In one embodiment, the present disclosure relates to a method for treating a subject with an ssRNA viral infection (e.g., Ebola virus or Marburg virus) by co-administering i) a therapeutically effective amount of an antiviral drug; and ii) a therapeutically effective amount of at least one anti-atypical mycobacterial agent. In one embodiment, the antiviral drug is brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU. Suitable anti-atypical mycobacterial agents include, but are not limited to, clarithromycin, rifabutin, rifampicin, azithromycin, roxithromycin, amikacin, clofazimine, ofloxacin ethambutol, ciprofloxacin, and oxazolidinones. In one embodiment, the anti-atypical mycobacterial agent is selected from at least one of rifabutin, clarithromycin, and clofazimine. In one embodiment, the anti-atypical mycobacterial agent is clofazimine. In one embodiment, the anti-atypical mycobacterial agent clofazimine is administered as a single solid oral dosage form.

In one embodiment, the present disclosure relates to a method of treating a subject with an ssRNA viral infection (e.g., Ebola virus) by administering in combination i) a therapeutically effective amount of an antiviral drug, ii) a therapeutically effective amount of rifabutin; iii) a therapeutically effective amount of clofazimine; and iv) a therapeutically effective amount of clarithromycin. In one embodiment, the antiviral drug is brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU.

In one embodiment, the present disclosure relates to a method of treating a subject having an ssRNA viral infection (such as, but not limited to, Ebola virus) by administering i) a therapeutically effective amount of an antiviral drug and ii) a therapeutically effective amount of clofazimine. In one embodiment, the antiviral drug is brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU.

In one embodiment, the present disclosure relates to a method of treating a subject with an ssRNA viral infection (such as, but not limited to, Ebola virus) by administering in combination i) a therapeutically effective amount of an antiviral drug; ii) a therapeutically effective amount of rifabutin; and iii) a therapeutically effective amount of clofazimine. In one embodiment, the antiviral drug is brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU. In one embodiment, rifabutin and clofazimine are administered as a single solid oral dosage form. In one embodiment, rifabutin and clofazimine are administered as separate solid oral dosage forms.

In one embodiment, the present disclosure relates to a method of treating a subject having an ssRNA viral infection, such as, but not limited to, Ebola virus, by administering in combination i) a therapeutically effective amount of brivudine, an active metabolite of brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU, and ii) a therapeutically effective amount of clarithromycin as an intravenous infusion.

In one embodiment, the present disclosure relates to a method of treating a subject suffering from an ssRNA viral infection (such as, but not limited to, Ebola virus) by administering a therapeutically effective amount of an aryl adamantane compound. In one embodiment, the aryl adamantane compound is selected from a compound of formula 7.

In one embodiment, the present disclosure relates to a method for treating a subject with an ssRNA viral infection (such as, but not limited to, Ebola virus) by administering in combination i) a therapeutically effective amount of brivudine, an active metabolite of brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU, and ii) a therapeutically effective amount of an aryl adamantane compound. In one embodiment, the aryl adamantane compound is selected from compounds of Formula 7.

In one embodiment, the present disclosure relates to a method for treating a subject with an ssRNA viral infection (e.g., Ebola virus or Marburg virus) by co-administering i) a therapeutically effective amount of an antiviral drug; ii) a therapeutically effective amount of at least one anti-atypical mycobacterial agent; and iii) a therapeutically effective amount of an aromatic adamantane compound. In one embodiment, the antiviral drug is brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or BVDU in a protected or prodrug form. Suitable anti-atypical mycobacterial agents include, but are not limited to, clarithromycin, rifabutin, rifampicin, azithromycin, roxithromycin, amikacin, clofazimine, ofloxacin ethambutol, ciprofloxacin, and oxazolidinones. In one embodiment, the anti-atypical mycobacterial agent is selected from at least one of rifabutin, clarithromycin, and clofazimine. In one embodiment, the anti-atypical mycobacterial agent is clofazimine. In one embodiment, the anti-atypical mycobacterial drug clofazimine is administered as a single solid oral dosage form. In one embodiment, the aromatic adamantane compound is selected from compounds of Formula 7.

In one embodiment, a treatment for an ssRNA viral infection (e.g., Ebola virus) comprises providing intravenous fluids (IV) and balanced electrolytes (body salts) to the subject; maintaining the subject's oxygen status and blood pressure; and administering in combination i) a therapeutically effective amount of an antiviral drug; ii) a therapeutically effective amount of rifabutin; iii) a therapeutically effective amount of clofazimine; and iv) a therapeutically effective amount of clarithromycin. In one embodiment, the antiviral drug is brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU.

In one embodiment, treatment for an ssRNA viral infection (such as, but not limited to, Ebola virus) comprises providing intravenous fluids (IV) and balanced electrolytes (body salts) to the subject; maintaining the subject's oxygen status and blood pressure; and administering i) a therapeutically effective amount of an antiviral drug; and ii) a therapeutically effective amount of clofazimine. In one embodiment, the antiviral drug is brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU.

In one embodiment, treatment for an ssRNA viral infection (such as, but not limited to, Ebola virus) comprises providing intravenous fluids (IV) and balanced electrolytes (body salts) to the subject; maintaining the subject's oxygen status and blood pressure; and administering in combination i) a therapeutically effective amount of an antiviral drug; ii) a therapeutically effective amount of clofazimine; and iii) a therapeutically effective amount of rifabutin. In one embodiment, the antiviral drug is brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU. In one embodiment, rifabutin and clofazimine are administered as a single solid oral dosage form. In one embodiment, rifabutin and clofazimine are administered as separate solid oral dosage forms.

In one embodiment, treatment for ssRNA viral infection (such as, but not limited to, Ebola virus) comprises providing intravenous fluids (IV) and balanced electrolytes (body salts) to the subject; maintaining the subject's oxygen status and blood pressure; and administering in combination i) a therapeutically effective amount of an antiviral drug; and ii) a therapeutically effective amount of clarithromycin as an intravenous infusion. In one embodiment, the antiviral drug is brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU.

In one embodiment, treatment for ssRNA viral infection (such as, but not limited to, Ebola virus) comprises providing intravenous fluids (IV) and balanced electrolytes (body salts) to the subject; maintaining the subject's oxygen status and blood pressure; and administering a therapeutically effective amount of an aryl adamantane compound. In one embodiment, the aryl adamantane compound is selected from compounds of Formula 7.

In one embodiment, treatment for ssRNA viral infection (such as, but not limited to, Ebola virus) comprises providing intravenous fluids (IV) and balanced electrolytes (body salts) to the subject; maintaining the subject's oxygen status and blood pressure; and administering in combination i) a therapeutically effective amount of an antiviral drug; and ii) a therapeutically effective amount of at least one anti-atypical mycobacterial agent. In one embodiment, the antiviral drug is brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU. Suitable anti-atypical mycobacterial agents include, but are not limited to, clarithromycin, rifabutin, rifampicin, azithromycin, roxithromycin, amikacin, clofazimine, ofloxacin ethambutol, ciprofloxacin, and oxazolidinones. In one embodiment, the anti-atypical mycobacterial agent is selected from at least one of rifabutin, clarithromycin, and clofazimine. In one embodiment, the anti-atypical mycobacterial agent is clofazimine. In one embodiment, the anti-atypical mycobacterial drug clofazimine is administered as a single solid oral dosage form.

In one embodiment, treatment for ssRNA viral infection (such as, but not limited to, Ebola virus) comprises providing intravenous fluids (IV) and balanced electrolytes (body salts) to the subject; maintaining the subject's oxygen status and blood pressure; and administering in combination i) a therapeutically effective amount of an antiviral drug; ii) a therapeutically effective amount of at least one anti-atypical mycobacterial agent; and iii) a therapeutically effective amount of an aromatic adamantane compound. In one embodiment, the antiviral drug is brivudine (BVDU), an active metabolite of BVDU, a salt thereof, or a protected or prodrug form of BVDU. Suitable anti-atypical mycobacterial agents include, but are not limited to, clarithromycin, rifabutin, rifampicin, azithromycin, roxithromycin, amikacin, clofazimine, ofloxacin ethambutol, ciprofloxacin, and oxazolidinones. In one embodiment, the anti-atypical mycobacterial agent is selected from at least one of rifabutin, clarithromycin, and clofazimine. In one embodiment, the anti-atypical mycobacterial agent is clofazimine. In one embodiment, the anti-atypical mycobacterial drug clofazimine is administered as a single solid oral dosage form. In one embodiment, the aryl adamantane compound is selected from compounds of Formula 7.

If necessary, the compounds of the present invention can be used in mechanical tests, using the assay method generally known in the prior art to determine whether other combinations or single agents and the combination of the present invention are equally effective in suppressing viral diseases (for example, those described herein). For example, candidate compounds can be detected alone or in combination (for example, with agents that inhibit viral replication, such as agents described herein) and applied to cells (for example, hepatocytes such as HepG2, renal epithelial cells such as 293T, macrophages such as THP-1 or isolated primary cells). After the appropriate time, the viral replication or load of these cells are checked. The reduction of viral replication or viral load identifies candidate compounds or reagent combinations as effective agents for treating viral diseases.

The compositions and methods of the present invention can include compounds or preparations that produce the concentration of the compound for treating a filovirus-mediated disease when applied to a subject. Compounds can be included in any suitable carrier material in any suitable amount, and are typically present in an amount of 1-95% by weight of a composition's total weight. Compositions can be provided in the form of oral, parenteral (e.g., intravenous or intramuscular), rectal, dermatological, skin, nose, vagina, inhalant, skin (patch), eyes, intrathecal or intracranial administration routes. Therefore, compositions can be, for example, tablets, capsules, pills, powders, granules, suspensions, emulsions, solutions, gels comprising hydrogels, pastes, ointments, creams, plasters, water-permeable agents, osmotic delivery devices, suppositories, enemas, injections, implants, sprays or aerosols. Pharmaceutical compositions can be prepared according to conventional pharmaceutical practice.

The pharmaceutical compositions according to the present invention or for use in the methods of the present invention can be formulated to release the active compound immediately after administration or to release the active compound at any predetermined time or time period after administration. The latter type of composition is generally referred to as a controlled release formulation, which includes (i) formulations that produce a substantially constant concentration of the agent of the present invention in the body over an extended period of time; (ii) formulations that produce a substantially constant concentration of the agent of the present invention in the body for a long time after a predetermined lag time; (iii) formulations that maintain the effect of the agent for a predetermined period of time by maintaining a relatively constant effective dose of the agent in the body while minimizing undesirable side effects associated with fluctuations in the plasma level of the agent (sawtooth kinetic pattern); (iv) formulations that localize the effect of the agent, for example, by spatially arranging the controlled release composition adjacent to or within a diseased tissue or organ; (v) formulations that achieve ease of administration, for example, administration once a week or once every two weeks; and (vi) formulations that achieve targeted action of the agent by using a carrier or chemical derivative to deliver the combination to a specific target cell type.

Any number of strategies can be employed to achieve controlled release, wherein the release rate exceeds the metabolic rate of the compound. In one embodiment, controlled release is achieved by appropriate selection of various formulation parameters and ingredients, including, for example, various types of controlled release compositions and coatings. Thus, the compound is formulated with appropriate excipients into a pharmaceutical composition that releases the compound in a controlled manner upon administration. Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, molecular complexes, microspheres, nanoparticles, patches, and liposomes.

It is not desirable to limit the administration of the compounds to a single formulation and method of delivery for all compounds in the combination. The combination can be administered using separate formulations and/or methods of delivery for each compound using, for example, a combination of any of the above formulations and methods. In one example, a first agent is delivered orally and a second agent is delivered intravenously.

The dosage of the compound or combination of compounds depends on several factors, including: the method of administration, the type of disease to be treated, the severity of the infection, whether it occurs first in the early or late stages of the infection, and the age, weight, and health status of the patient to be treated. For combinations including the synergistic pairs of agents identified herein, the recommended dosage of the antiviral agent may be less than or equal to the recommended dosage given in the Physician's Desk Reference, 69th Edition (2015).

As mentioned above, the compound in question can be administered in the form of tablets, capsules, elixirs or syrups, or rectally in the form of suppositories. Parenteral administration of the compound is suitably carried out in the form of, for example, saline solutions or compounds incorporated into liposomes. In cases where the compound itself is not easily soluble, a solubilizing agent such as ethanol can be used. The correct dosage of the compound can be determined by examining its efficacy in viral replication assays and its toxicity in humans.

The medicament of the present invention is also a useful tool for illustrating the mechanism information of the biological pathways related to viral diseases. This information can cause the development of new combinations or single agents for treating, preventing or alleviating viral diseases. Methods known in the art for determining biological pathways can be used to determine the pathway or pathway network affected by contacting cells infected with viruses (e.g., primary macrophages) with the compounds of this invention. These methods can include, compared with untreated, positive or negative control compounds and/or new single agents and combinations, analyzing the cellular components expressed or suppressed after contact with the compounds of this invention, or analyzing some other activities of cells or viruses (such as enzyme activity, nutrient absorption and proliferation). The cellular components analyzed can include gene transcripts and protein expression. Suitable methods can include standard biochemical techniques, radiolabeled compounds of the present invention (e.g., ¹⁴ C or ³ H labels), and observing protein-bound compounds, such as using 2D gels, gene expression profiles. Once identified, such compounds can be used in vivo models (e.g., knockout or transgenic mice) to further verify the tool or develop new agents or strategies for the treatment of viral diseases.

Example

The following examples are intended to illustrate but not to limit the present invention.

Example 1 : In vitro screening experiment

Screening experiments provided results regarding measurable inhibition of ssRNA virus replication in cell culture using in vitro plaque assays.

For the assay, primary human macrophages were seeded at 1×10 ⁵ cells/well in 100 μl of 10% RPMI 1640 medium (96-well plates coated with Biocoat I collagen I). The next day, the medium was removed and 100 μl of medium containing different concentrations of drug was added for one (1) hour. Afterwards, 15 μl of virus-medium mixture was added to each well for one (1) hour (multiplicity of infection (MOI) was 0.01, 0.1 or 1.0). The virus inoculum was removed and 100 μl of medium containing different concentrations of drug was added. The plates were kept in a 37°C incubator for 48 hours. After 48 hours of infection, the supernatant was collected for plaque assay and the plates were fixed with 10% NBF.7. The plates were then stained with anti-EBOV VP40. The plates were washed three (3) times with PBS to remove NBF. The cells were permeabilized with 0.25% Triton X-100 in PBS for 5 minutes at room temperature. The plates were washed three (3) times with PBS and then blocked with 10% BSA in PBS for 30 minutes at 37°C. The cells were stained with primary antibody (anti-EBOVVp40 BMD004B007) at 1:1000 in 3% BSA (PBS) at 37°C for 2 hours. The plates were washed three (3) times with PBS again and stained with secondary antibody (Alex-488 conjugated goat anti-mouse IgG) at 1:2000 in 3% BSA (PBS) at 37°C for 45 minutes. The plates were washed three (3) times with PBS again and the cells were stained with Horst for 10 minutes at room temperature. The plates were washed three (3) times with PBS and then scanned using a high-content imaging system. The percentage of infected cells was measured and analyzed using a high-content imaging system with high-content imaging and analysis software.

Primary human macrophages were seeded at 1×10 ⁵ cells/well in 100 μl of 10% RPMI 1640 medium (96-well plates coated with Biocoat type I collagen). The culture medium was removed the next day and 100 μl of culture medium containing different concentrations of drug was added. 48 h after treatment, cytotoxicity was measured using Dojindo's Cell Counting Kit-8 (CCK-8), which uses Dojindo's highly water-soluble tetrazolium salt (WST). WST-8 produces a water-soluble formazan dye when reduced in the presence of an electron mediator. Viability was calculated relative to the untreated cell control, which was set to 100% viability.

The assay described above was used to identify drugs that inhibit Ebola virus replication in primary human macrophages.

The table below lists the concentrations of tested drugs and drug combinations and their effects on toxicity in primary human macrophages.

Based on these results, specific concentrations and combinations of drugs were tested to determine whether they could inhibit Ebola virus replication.

The table below provides a summary of the concentrations of drugs and drug combinations that were found to be both non-toxic and capable of reducing Ebola virus infection of primary macrophages.

Clarithromycin alone can inhibit EBOV infection in primary macrophages. When rifabutin and clarithromycin are administered together, there appears to be a synergistic effect.

Prophetic Embodiments

Example 2 : In vivo screening experiment - Ebola - non-human primates

Use macaques to determine whether the administration of the medicament of the present invention or the medicament combination of the present invention for a suitable time period can inhibit viral replication, reduce viral load or alleviate at least one symptom associated with Ebola virus. The experiment will be composed of the same number of non-human primates (NHPs) in each study group, and each study group will receive various doses of the medicament of the present invention alone and together for a suitable time period of various doses of the medicament of the present invention, starting at least one day after a lethal dose of intramuscular challenge, such as 4,000 × median tissue culture infection dose (TCID50) of EBOV-K (or 2,512 plaque forming units (pfu). It is planned that the NHPs in each study group will be divided into different groups and receive the one or more medicaments at different time points after infection, such as 30 to 75 minutes (Group 1), 24 hours (Group 2), 48 hours (Group 3) and 72 hours (Group 4) after infection), and will include at least once a day. Control animals consisting of the same number of NHPs as the study group will be given phosphate buffered saline (PBS). Kaplan-Meier survival curves will be created for each group, and clinical scores and fever (rectal temperature) will be measured. In addition, viral titers (EBOC viremia) will be measured by TCID 50. Blood cell counts and serum biochemistry will be measured, including but not limited to white blood cell count, lymphocyte count, lymphocyte percentage, platelet count, neutrophil count, neutrophil percentage, alanine aminotransferase, alkaline phosphatase, blood urea nitrogen, creatinine, and glucose.

Example 3 : In vivo screening experiment - Marburg - non-human primates

Macaque will be used to determine whether the appropriate time period for administering medicament of the present invention or medicament combination of the present invention can suppress viral replication, reduce viral load or alleviate at least one symptom relevant to Marburg virus.The experiment will be made up of the non-human primates (NHP) of the same number in each research group, and each research group accepts the reagent of the present invention of various dosage separately and accepts the appropriate time period of reagent of the present invention of various dosage together, starting at least one day after lethal dose MARV-Angola.Expectation will be divided into different groups by the NHP from each research group, and it will receive reagent or multiple reagents at different time points after infection, for example 30 to 75 minutes (group 1), 24 hours (group 2), 48 hours (group 3) and 72 hours (group 4), and will comprise the dosage of at least one day.The control animal being made up of the NHP of the same number with research group will be given phosphate buffered saline (PBS).Kaplan-Meier survival curve will be created for every group, and clinical score and fever (rectal temperature) will be measured.In addition, by TCID50, measure viral titer. Blood counts and serum biochemistries will be measured, including but not limited to white blood cell count, lymphocyte count, lymphocyte percentage, platelet count, neutrophil count, neutrophil percentage, alanine aminotransferase, alkaline phosphatase, blood urea nitrogen, creatinine, and glucose.

or a pharmaceutically acceptable salt thereof, wherein:

_R1 is phenyl, 4-chlorophenyl or 4-fluorophenyl,

_R4 is H or alkyl, and

n is 1 or 2; and

In one embodiment, the effective amount of the compound of formula I is from about 15.0 mg/kg/day to about 20 mg/kg/day. In one embodiment, the determining step includes measuring viral load using a nucleic acid amplification-based test method at at least two different times within a suitable time period. In one embodiment, the inhibition of viral replication or the reduction of viral load determined using a nucleic acid amplification-based test method is at least 10%. In one embodiment, the individual is a human. In one embodiment, the filovirus is Ebola virus or Marburg virus. In one embodiment, the virus is Ebola virus. In one embodiment, the compound of formula I is present as a solid dosage form. In one embodiment, the solid dosage form is a capsule. In one embodiment, the method further comprises administering at least one antibiotic suitable for a time period to an individual infected or exposed to the filovirus, wherein the combination of the at least one antibiotic and the compound of formula I produces a synergistic effect. In one embodiment, the at least one antibiotic is selected from one of clarithromycin or rifabutin.

The methods of the present invention comprise administering at least two antibiotics to an individual infected or exposed to a virus, wherein the administering step is performed for a suitable period of time to allow the individual to be treated; and determining whether the individual has been treated, wherein the determining step comprises measuring one of: inhibition of viral replication, reduction in viral load, or alleviation of at least one symptom associated with the filovirus. In one embodiment, at least one antibiotic is a macrolide antibiotic. In one embodiment, at least one antibiotic is a rifamycin antibiotic. In one embodiment, the antibiotics are clarithromycin and rifabutin. In one embodiment, the effective amount of clarithromycin is from about 12.0 mg/kg/day to about 17.0 mg/kg/day. In one embodiment, the effective amount of rifabutin is from about 4.0 mg/kg/day to about 8.0 mg/kg/day. In one embodiment, the determining step comprises measuring viral load using a nucleic acid amplification-based assay at at least two different times over a suitable period of time. In one embodiment, the inhibition of viral replication or reduction in viral load as determined using a suitable assay is at least 10%. In one embodiment, the individual is a human. In one embodiment, the filovirus is Ebola virus or Marburg virus. In one embodiment, the filovirus is Ebola virus.

All patents, patent applications and open references cited herein are incorporated herein by reference in their entirety. Without departing from the scope and spirit of the present invention, various modifications and variations of the compositions and methods of the present invention will be apparent to those skilled in the art. Although the present invention has been described in conjunction with specific embodiments, it should be understood that the present invention should not be unduly limited to these specific embodiments. In fact, various modifications for implementing the described pattern of the present invention that are apparent to those skilled in the art in molecular biology, medicine, immunology, pharmacology, virology or related fields are intended to be within the scope of the present invention.

Treatment for inhibiting the replication of single-stranded RNA viruses

CROSS-REFERENCE TO RELATED APPLICATIONS

Background Art

Single-molecule negative-strand RNA viruses

○ Bornaviridae family - Borna disease virus

○ Filoviridae family - includes Ebola virus, Marburg virus

○ Rhabdoviridae family - includes rabies virus

Nyamiviridae family - includes niaviruses

Unclassified family:

Arenaviridae family – includes Lassa virus

Bunyaviridae family - includes Hantavirus, Crimean-Congo hemorrhagic fever

○Ophioviridae family

○ Orthomyxoviridae family - includes influenza viruses

Unclassified genus:

○Hepatitis D virus genus - including hepatitis E virus

○Dichlorovirus

○Emaravirus

Nivavirus – includes Niamani virus and Midway virus

○Tensivirus

○Meganevirus

Unclassified species:

Taastrup virus

○ Sclerotinia sclerotiorum negative-strand RNA virus 1

Summary of the Invention

or a pharmaceutically acceptable salt thereof, wherein:

_R1 is phenyl, 4-chlorophenyl or 4-fluorophenyl,

_R4 is H or alkyl, and

n is 1 or 2; and

DETAILED DESCRIPTION

Table 1

A non-limiting example of a prodrug form of BVDU is represented by Formula I:

Brivudine is represented by Formula 2:

Upamostat is represented by Formula 3:

and pharmaceutically acceptable salts thereof, wherein

L is a bond or -C(R ₃ ,R ₄ )-;

_Where )S(O) ₂ -;

The aryl adamantane compounds of formula 7 include compounds of formula I-1:

and pharmaceutically acceptable salts thereof, wherein:

Aryladamantane compounds of Formula 7 include those of Formula II:

and pharmaceutically acceptable salts thereof, wherein:

Y is -C(R ₄ ,R ₅ )-, -N(R ₄ )-, -O- or -C(O)-;

_R4 and _R5 are independently H or alkyl.

Compounds of Formula II include those wherein:

Y is -C(R ₄ ,R ₅ )- or -N(R ₄ )-;

_R3 is H, alkyl or oxo (=O); and

R ₄ and R ₅ are independently H or (C ₁ -C ₆ )alkyl.

Representative compounds of Formula II include:

Representative compounds of formula I-1 include:

and pharmaceutically acceptable salts thereof, wherein

○ _R1 is H, Cl or F;

○ _R2 is H or alkyl;

○m is 0, 1 or 2;

○n is 1, 2, 3, 4 or 5;

○Among them

● _R5 is H or alkyl,

● _R6 is H or alkyl, and

Each R ₇ is independently H or alkyl.

In certain embodiments of the compound of formula (I) as described above,

The part has the following structure

For example, compounds of the present invention include:

Example

Example 1 : In vitro screening experiment

Prophetic Embodiments

Example 2 : In vivo screening experiment - Ebola - non-human primates

Example 3 : In vivo screening experiment - Marburg - non-human primates

or a pharmaceutically acceptable salt thereof, wherein:

_R1 is phenyl, 4-chlorophenyl or 4-fluorophenyl,

_R4 is H or alkyl, and

n is 1 or 2; and

CROSS-REFERENCE TO RELATED APPLICATIONS

Background Art

Single-molecule negative-strand RNA viruses

○ Bornaviridae family - Borna disease virus

○ Filoviridae family - includes Ebola virus, Marburg virus

○ Rhabdoviridae family - includes rabies virus

Nyamiviridae family - includes niaviruses

Unclassified family:

Arenaviridae family – includes Lassa virus

Bunyaviridae family - includes Hantavirus, Crimean-Congo hemorrhagic fever

○Ophioviridae family

○ Orthomyxoviridae family - includes influenza viruses

Unclassified genus:

○Hepatitis D virus genus - including hepatitis E virus

○Dichlorovirus

○Emaravirus

Nivavirus – includes Niamani virus and Midway virus

○Tensivirus

○Meganevirus

Unclassified species:

Taastrup virus

○ Sclerotinia sclerotiorum negative-strand RNA virus 1

Summary of the Invention

or a pharmaceutically acceptable salt thereof, wherein:

_R1 is phenyl, 4-chlorophenyl or 4-fluorophenyl,

_R4 is H or alkyl, and

n is 1 or 2; and

DETAILED DESCRIPTION

Table 1

A non-limiting example of a prodrug form of BVDU is represented by Formula I:

Brivudine is represented by Formula 2:

Upamostat is represented by Formula 3:

and pharmaceutically acceptable salts thereof, wherein

L is a bond or -C(R ₃ ,R ₄ )-;

_Where )S(O) ₂ -;

The aryl adamantane compounds of formula 7 include compounds of formula I-1:

and pharmaceutically acceptable salts thereof, wherein:

Aryladamantane compounds of Formula 7 include those of Formula II:

and pharmaceutically acceptable salts thereof, wherein:

Y is -C(R ₄ ,R ₅ )-, -N(R ₄ )-, -O- or -C(O)-;

_R4 and _R5 are independently H or alkyl.

Compounds of Formula II include those wherein:

Y is -C(R ₄ ,R ₅ )- or -N(R ₄ )-;

_R3 is H, alkyl or oxo (=O); and

R ₄ and R ₅ are independently H or (C ₁ -C ₆ )alkyl.

Representative compounds of Formula II include:

Representative compounds of formula I-1 include:

and pharmaceutically acceptable salts thereof, wherein

○ _R1 is H, Cl or F;

○ _R2 is H or alkyl;

○m is 0, 1 or 2;

○n is 1, 2, 3, 4 or 5;

○Among them

● _R5 is H or alkyl,

● _R6 is H or alkyl, and

Each R ₇ is independently H or alkyl.

In certain embodiments of the compound of formula (I) as described above,

The part has the following structure

For example, compounds of the present invention include:

Example

Example 1 : In vitro screening experiment

Prophetic Embodiments

Example 2 : In vivo screening experiment - Ebola - non-human primates

Example 3 : In vivo screening experiment - Marburg - non-human primates

or a pharmaceutically acceptable salt thereof, wherein:

_R1 is phenyl, 4-chlorophenyl or 4-fluorophenyl,

_R4 is H or alkyl, and

n is 1 or 2; and

CROSS-REFERENCE TO RELATED APPLICATIONS

Background Art

Single-molecule negative-strand RNA viruses

○ Bornaviridae family - Borna disease virus

○ Filoviridae family - includes Ebola virus, Marburg virus

○ Rhabdoviridae family - includes rabies virus

Nyamiviridae family - includes niaviruses

Unclassified family:

Arenaviridae family – includes Lassa virus

Bunyaviridae family - includes Hantavirus, Crimean-Congo hemorrhagic fever

○Ophioviridae family

○ Orthomyxoviridae family - includes influenza viruses

Unclassified genus:

○Hepatitis D virus genus - including hepatitis E virus

○Dichlorovirus

○Emaravirus

Nivavirus – includes Niamani virus and Midway virus

○Tensivirus

○Meganevirus

Unclassified species:

Taastrup virus

○ Sclerotinia sclerotiorum negative-strand RNA virus 1

Summary of the Invention

or a pharmaceutically acceptable salt thereof, wherein:

_R1 is phenyl, 4-chlorophenyl or 4-fluorophenyl,

_R4 is H or alkyl, and

n is 1 or 2; and

DETAILED DESCRIPTION

Table 1

A non-limiting example of a prodrug form of BVDU is represented by Formula I:

Brivudine is represented by Formula 2:

Upamostat is represented by Formula 3:

and pharmaceutically acceptable salts thereof, wherein

L is a bond or -C(R ₃ ,R ₄ )-;

_Where )S(O) ₂ -;

The aryl adamantane compounds of formula 7 include compounds of formula I-1:

and pharmaceutically acceptable salts thereof, wherein:

Aryladamantane compounds of Formula 7 include those of Formula II:

and pharmaceutically acceptable salts thereof, wherein:

Y is -C(R ₄ ,R ₅ )-, -N(R ₄ )-, -O- or -C(O)-;

_R4 and _R5 are independently H or alkyl.

Compounds of Formula II include those wherein:

Y is -C(R ₄ ,R ₅ )- or -N(R ₄ )-;

_R3 is H, alkyl or oxo (=O); and

R ₄ and R ₅ are independently H or (C ₁ -C ₆ )alkyl.

Representative compounds of Formula II include:

Representative compounds of formula I-1 include:

and pharmaceutically acceptable salts thereof, wherein

○ _R1 is H, Cl or F;

○ _R2 is H or alkyl;

○m is 0, 1 or 2;

○n is 1, 2, 3, 4 or 5;

○Among them

● _R5 is H or alkyl,

● _R6 is H or alkyl, and

Each R ₇ is independently H or alkyl.

In certain embodiments of the compound of formula (I) as described above,

The part has the following structure

For example, compounds of the present invention include:

Example

Example 1 : In vitro screening experiment

Prophetic Embodiments

Example 2 : In vivo screening experiment - Ebola - non-human primates

Example 3 : In vivo screening experiment - Marburg - non-human primates

or a pharmaceutically acceptable salt thereof, wherein:

_R1 is phenyl, 4-chlorophenyl or 4-fluorophenyl,

_R4 is H or alkyl, and

n is 1 or 2; and

Claims

1. Use in the preparation of an effective amount of clarithromycin and an effective amount of rifabutin in the preparation of a medicament for treating an individual tested and identified as infected or exposed to a filovirus, wherein administration of said effective amount of clarithromycin and said effective amount of rifabutin results in inhibition of viral replication or reduction of viral load, and said administration is continued for an appropriate period of time so that the individual is treated.

2. The use according to claim 1, wherein the reduction in viral load is determined by using a nucleic acid amplification-based assay at two or more different times within the suitable time period.

3. The use according to claim 1, characterized in that the inhibition of viral replication or the reduction of viral load after treatment is at least 10%.

4. The use according to claim 1, wherein the individual is a person.

5. The use according to claim 1, wherein the filamentous virus is Ebola virus or Marburg virus.

6. The use according to claim 1, wherein the filamentous virus is an Ebola virus.

7. The use according to claim 1, characterized in that administration of the effective amount of clarithromycin and the effective amount of rifabutin results in the relief of at least one symptom associated with the filovirus.

8. The use according to claim 1, characterized in that the administration of the effective amount of clarithromycin and the effective amount of rifabutin results in synergistic inhibition of viral replication or synergistic reduction of viral load.