WO2022113069A1

WO2022113069A1 - E protein channel blockers and orf3 inhibitors as anti-covid-19 agents

Info

Publication number: WO2022113069A1
Application number: PCT/IL2021/051396
Authority: WO
Inventors: Isaiah Arkin; Prabhat Pratap SINGH TOMAR; Anamika Singh; Miriam KRUGLIAK
Original assignee: Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd.
Priority date: 2020-11-24
Filing date: 2021-11-24
Publication date: 2022-06-02
Also published as: WO2022113069A9

Abstract

The present invention is directed to pharmaceutical compositions including a SARS- CoV-2 E protein channel blocker and ORF3 inhibitor, and methods of using same, such as for treating or preventing SARS-CoV-2 virulence in a subject in need thereof.

Description

E PROTEIN CHANNEL BLOCKERS AND ORF3 INHIBITORS AS ANTI-COVID-

19 AGENTS

CROSS-REFERENCE TO RELATED APPLICATIONS

[001] This application claims the benefit of priority of U.S. Provisional Patent Application No. 63/117,619, titled "E PROTEIN CHANNEL BLOCKERS AND ORF3 INHIBITORS AS ANTI-COVID- 19 AGENTS", filed November 24, 2020, and of U.S. Provisional Patent Application No. 63/274,979, titled "E PROTEIN CHANNEL BLOCKERS AND ORF3 INHIBITORS AS ANTI-COVID-19 AGENTS", filed November 3, 2021. The contents of both applications are incorporated herein by reference in their entirety.

FIELD OF INVENTION

[002] The present invention is in the field of anti- viral therapy.

BACKGROUND

[003] Coronaviruses are positive-sense, single-stranded RNA viruses that are often associated with mild respiratory tract infections in humans. However, three members of the family have received notoriety due to their abnormal virulence: SARS-CoV-1 was the etiological agent of the SARS epidemic in the winter of 2002/3 that caused 774 deaths amongst 8,098 cases; MERS-CoV was responsible for the MERS epidemic that started from 2012 with 862 deaths from 2506 infections; Finally, SARS-CoV-2 is responsible for the ongoing COVID-2019 pandemic resulting in 4,997,967 deaths out of 246,535,404 cases (as of Sunday Oct 31, 2021).

[004] Genomic analyses have indicated that SARS-CoV-1 and SARS-CoV-2 are very similar to one another (ca. 80%) but are distinct from most other Coronaviridae members that infect humans. Both viruses have been placed in subgroup B in the Betacoronavirus genus within the Orthocoronavirinae subfamily of the Coronaviridae.

[005] Of all coronavirus’ structural proteins, E is the least understood in terms of mechanism of action and structure. Functionally, the E protein has been implicated in viral assembly, release, and pathogenesis. Yet crucially, coronavirus E proteins are important for viral pathogenesis, and attenuated viruses lacking the protein have even been suggested to serve as vaccine candidates. Co V-2 3a protein, also known as open reading implicated in assembly of homotetrameric potassium sensitive ion channels (viroporin) and may modulate virus release. Additionally, it is implicated in pathogenesis, including up- regulation of expression of fibrinogen subunits FGA, FGB and FGG in host lung epithelial cells, inducement of apoptosis in cell culture.

[007] There is still a great need for synergistic combinations of any one of 3 a protein inhibitors, E protein inhibitors, and both, for SARS-CoV-2 prevention and/or treatment.

SUMMARY

[008] According to a first aspect, there is provided a pharmaceutical composition comprising a SARS-CoV-23a protein inhibitor and a SARS-CoV-2 E channel blocker.

[009] According to another aspect, there is provided a pharmaceutical composition comprising Flumatinib and Darapladib.

[010] According to another aspect, there is provided a combination of a SARS-CoV-2 3a protein inhibitor and a SARS-CoV-2 E channel blocker, for use in the treatment or prevention of SARS-CoV-2 virulence in a subject in need thereof, wherein the SARS-CoV- 2 3a protein inhibitor and a SARS-CoV-2 E channel blocker are provided at a molar per molar ratio ranging from 10:1 to 1:10 to the subject.

[011] According to another aspect, there is provided a combination of Flumatinib and Darapladib, for use in the treatment or prevention of SARS-CoV-2 virulence in a subject in need thereof.

[012] According to another aspect, there is provided a method for treating or preventing SARS-CoV-2 virulence in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a SARS-CoV-23a protein inhibitor and a SARS-CoV-2 E channel blocker, thereby treating or preventing SARS-CoV-2 virulence in the subject.

[013] According to another aspect, there is provided a method for treating or preventing SARS-CoV-2 virulence in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Flumatinib and Darapladib.

[014] In some embodiments, the SARS-CoV-2 protein 3a inhibitor is selected from Flumatinib or Darapladib. [015] In some embodiments, the SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Saroglitazar, Mebrofenin, Cyclen, Plerixafor, and

Gliclazide.

[016] In some embodiments, the SARS-CoV-2 protein 3a inhibitor is Flumatinib and the SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Saroglitazar, Mebrofenin, Cyclen, Plerixafor, and Gliclazide.

[017] In some embodiments, the SARS-CoV-2 protein 3a inhibitor is Darapladib and the SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Mebrofenin, and Cyclen.

[018] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[019] In some embodiments, the pharmaceutical composition is for use in treatment or prevention of SARS-CoV-2 virulence in a subject in need thereof.

[020] In some embodiments, the SARS-CoV-2 3a protein inhibitor is formulated within a first pharmaceutical composition and the SARS-CoV-2 E channel blocker is formulated within a second pharmaceutical composition.

[021] T In some embodiments, the prevention or preventing comprises prevention or preventing of any one of: SARS-CoV-2 entry to a cell of the subject, uncoating of the SARS- CoV-2, release of the SARS-CoV-2 from a cell of the subject, or any combination thereof.

[022] In some embodiments, the subject is infected or suspected of being infected by SARS-CoV-2.

[023] Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

BRIEF DESCRIPTION OF THE FIGURES

[024] Fig. 1 includes a graph showing the effect of individual 3 a protein inhibitors on cell survival. Both Flumatinib and Darapladib provided near full protection at a concentration of 3 μM. include graphs showing the effect of the inhibitor and various E protein inhibitors on cell survival. (2A) Combination of Flumatinib and Mavorixafor; (2B) Combination of Flumatinib and Cyclen; and (2C) Combination of Flumatinib and Sargolitazar.

[026] Figs. 3A-3C include graphs showing the effect of a combination of 3a protein inhibitors on cell survival. Combinations of Flumatinib and Darapladib were tested. (3A) Flumatinib (1 μM) and Darapladib (0.3 μM); (3B) Flumatinib (0.1 μM) and Darapladib (1 μM); and (3C) Flumatinib (0.3 μM) and Darapladib (1 μM).

[027] Figs. 4A-4C include graphs showing the effect of 3a inhibitors, E protein blockers, or both, on cell survival. (4A) The sole effect of 3 μM of either 3a inhibitors or E protein blockers; (4B) Combinations of 3μM of the E protein blockers: Mavorixafor (Mavo), Saroglitazar (Saro), Mebrofenin (Mebro), Cyclen, 5-Azacytidine (5-aza), Plerixafor (Plerixa), Kasugamycin (Kasuga), and Gliclazide (glicla), with the 3a protein inhibitors: Capreomycin (Capero), Pentamidine (Penta), Spectinomycin (Spectino), and Flumatinib (Fluma); (4C) Combinations of 3μM of the E protein blockers: Mavo, Saro, Mebro, Cyclen, 5-aza, Plerixa, Kasuga, and glicla, with the 3a protein inhibitors: Fludarabine (Fluda), Fitronesib (Fitro), Darapladib (Darap), and Floxuridine (Fluxo).

DETAILED DESCRIPTION

[028] The present invention, in some embodiments, provides compositions comprising a SARS-CoV-2 E protein channel blocker and a SARS-CoV-23a protein inhibitor for treating or preventing SARS-CoV-2 virulence in a subject. The present invention, in some embodiments, provides compositions comprising a plurality of SARS-CoV-2 3a protein inhibitors for treating or preventing SARS-CoV-2 virulence in a subject. The present invention, in some embodiments, provides compositions comprising a SARS-CoV-2 E protein channel blocker and a SARS-CoV-23a protein inhibitor, for preventing SARS-CoV- 2 cell entry, uncoating and/or release from a cell. The present invention, in some embodiments, provides compositions comprising a plurality of SARS-CoV-2 3a protein inhibitors, for preventing SARS-CoV-2 cell entry, uncoating and/or release from a cell.

[029] SARS-CoV-2 E protein is known to one skilled in the art and has a GenBank Accession no: QIH45055.1. According to some embodiments, the SARS-CoV-2 E protein comprises the amino acid sequence as set forth in SEQ ID NO 1:

According to some embodiments, comprises an analog of SEQ ID NO: 1, such as an analog having at least 85%, at least 90%, at least 95% identity to SEQ ID NO: 1.

[030] According to some embodiments, the invention provides a method of treating or preventing SARS-CoV-2 virulence in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a SARS-CoV-2 E protein channel blocker and a SARS-CoV-2 3a protein inhibitor, thereby treating or preventing SARS-CoV-2 virulence in the subject.

[031] In some embodiments, administering comprises providing a SARS-CoV-2 E protein channel blocker and a SARS-CoV-2 3a protein in a therapeutically effective amount, synergistically effective amount, or both, to the subject.

[032] In some embodiments, administering comprises providing at least two SARS-CoV- 2 3 a protein inhibitors in a therapeutically effective amount, synergistically effective amount, or both, to the subject.

[033] As used herein, the term “synergistically effective amount” comprises any weight or concentration of a SARS-CoV-2 E protein channel blocker and a SARS-CoV-2 3a protein inhibitor, as long as their molar ratio ranges from: 100: 1 to 1 : 100 (M:M), 50: 1 to 1 :50 (M:M), 30:1 to 1:30 (M:M), 10: lo 1:10 (M:M), 8:1 to 1:8 (M:M), 5:1 to 1:5 (M:M), 4:1 to 1:4 (M:M), 3:1 to 1:3 (M:M), 2:1 to 1:2 (M:M), or is 1:1 (M:M). Each possibility represents a separate embodiment of the invention.

[034] As used herein, the term “synergistically effective amount” comprises any weight or concentration of a first SARS-CoV-2 3a protein channel inhibitor and a second SARS-CoV- 2 3a protein inhibitor, as long as their molar ratio ranges from: 100:1 to 1:100 (M:M), 50:1 to 1:50 (M:M), 30:1 to 1:30 (M:M), 10:1 to 1:10 (M:M), 8:1 to 1:8 (M:M), 5:1 to 1:5 (M:M), 4:1 to 1:4 (M:M), 3:1 to 1:3 (M:M), 2:1 to 1:2 (M:M), or is 1:1 (M:M). Each possibility represents a separate embodiment of the invention.

[035] In some embodiments, the first 3a protein inhibitor comprises any one of Flumatinib and Darapladib. In some embodiments, the second 3a protein inhibitor comprises any one of Flumatinib and Darapladib.

[036] According to some embodiments, the invention provides a method of treating or preventing SARS-CoV-2 virulence in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a combination of at least 2 SARS-CoV-2 3a protein inhibitors, thereby treating or preventing SARS-CoV-2 virulence in the subject. embodiments, the invention provides a method Coronavirus disease 2019 (COVID-19) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a SARS-CoV-2 E protein channel blocker and a SARS-CoV-2 3a protein inhibitor, thereby treating or preventing COVID-19.

[038] In some embodiments, the invention provides a method of treating or preventing Coronavirus disease 2019 (COVID-19) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a combination of at least two SARS-CoV-2 3a protein inhibitors, thereby treating or preventing COVID-19.

[039] According to some embodiments, the invention provides a method of preventing SARS-CoV-2 release from a cell. In some embodiments, the method comprises contacting a cell with a SARS-CoV-2 E protein channel blocker and a SARS-CoV-2 3a protein inhibitor, thereby preventing SARS-CoV-2 release from the cell. In some embodiments, the method comprises contacting a cell with at least 2 SARS-CoV-2 3a protein inhibitors, thereby preventing SARS-CoV-2 release from the cell.

[040] According to some embodiments, the invention provides a method of preventing SARS-CoV-2 cell entry. In some embodiments, the method comprises contacting a cell with a SARS-CoV-2 E protein channel blocker and a SARS-CoV-2 3a protein inhibitor, thereby preventing SARS-CoV-2 cell entry. In some embodiments, the method comprises contacting a cell with at least 2 SARS-CoV-2 3a protein inhibitors, thereby preventing SARS-CoV-2 cell entry.

[041] According to some embodiments, the invention provides a method of preventing SARS-CoV-2 uncoating. In some embodiments, the method comprises contacting a cell with a SARS-CoV-2 E protein channel blocker and a SARS-CoV-2 3a protein inhibitor, thereby preventing SARS-CoV-2 uncoating. In some embodiments, the method comprises contacting a cell with at least 2 SARS-CoV-2 3a protein inhibitors, thereby preventing SARS-CoV-2 uncoating.

[042] According to some embodiments, a cell is a cell of a subject. According to some embodiments, contacting comprises administering to the subject. According to some embodiments, the subject is a subject infected or suspected as being infected by SARS-CoV-

2.

[043] According to some embodiments, there is provided a method for treating or preventing SARS-CoV-2 virulence in a subject in need thereof, the method comprising to the subject a therapeutically effective amount composition comprising a SARS-CoV-2 3a protein inhibitor and a SARS-CoV-2 E channel blocker in a molar per molar ratio ranging from 100:1 to 1:100, 50:1 to 1:50, 30:1 to 1:30, 10:1 to 1:10, 8:1 to 1:8, 5:1 to 1:5, 4:1 to 1:4, 3:1 to 1:3, 2:1 to 1:2, or is 1:1, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[044] In some embodiments, the SARS-CoV-2 protein 3a inhibitor is Flumatinib or Darapladib.

[045] In some embodiments, the SARS-CoV-2 E channel blocker is selected: Mavorixafor, Saroglitazar, Mebrofenin, Cyclen, Plerixafor, and Gliclazide.

[046] In some embodiments, the SARS-CoV-2 protein 3a inhibitor is Flumatinib and the SARS-CoV-2 E channel blocker is selected from: Mavorixafor, Saroglitazar, Mebrofenin, Cyclen, Plerixafor, and Gliclazide.

[047] In some embodiments, the SARS-CoV-2 protein 3a inhibitor is in some embodiments, the SARS-CoV-2 protein 3a inhibitor is Flumatinib or Darapladib and the SARS-CoV-2 E channel blocker is selected from: Mavorixafor, Cyclen, or Mebrofenin.

[048] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Flumatinib and Mavorixafor, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[049] In some embodiments, the method comprises administering to the subject a synergistically effective amount of a pharmaceutical composition comprising Flumatinib and Mavorixafor, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[050] In some embodiments, the method comprises administering to the subject a pharmaceutical composition comprising Flumatinib and Mavorixafor in a molar per molar ratio ranging from 100:1 to 1:100, 50:1 to 1:50, 30:1 to 1:30, 15:1 to 1:15, 10:1 to 1:10, 8:1 to 1:8, 4:1 to 1:4, 3:1 to 1:3, 2:1 to 1:2, or is 1:1, thereby treating or preventing SARS-CoV- 2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[051] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Flumatinib treating or preventing SARS-CoV-2 possibility represents a separate embodiment of the invention.

[052] In some embodiments, the method comprises administering to the subject a synergistically effective amount of a pharmaceutical composition comprising Flumatinib and Cyclen, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[053] In some embodiments, the method comprises administering to the subject a pharmaceutical composition comprising Flumatinib and Cyclen in a molar per molar ratio ranging from100:1 to 1:100, 50:1 to 1:50, 30:1 to 1:30, 15:1 to 1:15, 10:1 to 1:10, 8:1 to 1:8, 4:1 to 1:4, 3:1 to 1:3, 2:1 to 1:2, or is 1:1, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[054] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Flumatinib and Mebrofenin, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[055] In some embodiments, the method comprises administering to the subject a synergistically effective amount of a pharmaceutical composition comprising Flumatinib and Mebrofenin, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[056] In some embodiments, the method comprises administering to the subject a pharmaceutical composition comprising Flumatinib and Mebrofenin in a molar per molar ratio ranging from 100:1 to 1:100, 50:1 to 1:50, 30:1 to 1:30, 15:1 to 1:15, 10:1 to 1:10, 8:1 to 1:8, 4:1 to 1:4, 3:1 to 1:3, 2:1 to 1:2, or is 1:1, thereby treating or preventing SARS-CoV- 2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[057] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Flumatinib and Saroglitazar, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[058] In some embodiments, the method comprises administering to the subject a synergistically effective amount of a pharmaceutical composition comprising Flumatinib and Saroglitazar, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention. embodiments, the method comprises pharmaceutical composition comprising Flumatinib and Saroglitazar in a molar per molar ratio ranging from 100:1 to 1:100, 50:1 to 1:50, 30:1 to 1:30, 15:1 to 1:15, 10:1 to 1:10, 8:1 to 1:8, 4:1 to 1:4, 3:1 to 1:3, 2:1 to 1:2, or is 1:1, thereby treating or preventing SARS-CoV- 2 virulence in the subject. Each possibility represents a separate embodiment of the invention. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Flumatinib and Plerixafor, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[060] In some embodiments, the method comprises administering to the subject a synergistically effective amount of a pharmaceutical composition comprising Flumatinib and Plerixafor, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[061] In some embodiments, the method comprises administering to the subject a pharmaceutical composition comprising Flumatinib and Plerixafor in a molar per molar ratio ranging from 100:1 to 1:100, 50:1 to 1:50, 30:1 to 1:30, 15:1 to 1:15, 10:1 to 1:10, 8:1 to 1:8, 4:1 to 1:4, 3:1 to 1:3, 2:1 to 1:2, or is 1:1, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[062] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Flumatinib and Gliclazide, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[063] In some embodiments, the method comprises administering to the subject a synergistically effective amount of a pharmaceutical composition comprising Flumatinib and Gliclazide, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[064] In some embodiments, the method comprises administering to the subject a pharmaceutical composition comprising Flumatinib and Gliclazide in a molar per molar ratio ranging from 100:1 to 1:100, 50:1 to 1:50, 30:1 to 1:30, 15:1 to 1:15, 10:1 to 1:10, 8:1 to 1:8, 4:1 to 1:4, 3:1 to 1:3, 2:1 to 1:2, or is 1:1, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[065] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Darapladib thereby treating or preventing SARS-CoV-2 possibility represents a separate embodiment of the invention.

[066] In some embodiments, the method comprises administering to the subject a synergistically effective amount of a pharmaceutical composition comprising Darapladib and Mavorixafor, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[067] In some embodiments, the method comprises administering to the subject a pharmaceutical composition comprising Darapladib and Mavorixafor in a molar per molar ratio ranging from 100:1 to 1:100, 50:1 to 1:50, 30:1 to 1:30, 15:1 to 1:15, 10:1 to 1:10, 8:1 to 1:8, 4:1 to 1:4, 3:1 to 1:3, 2:1 to 1:2, or is 1:1, thereby treating or preventing SARS-CoV- 2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[068] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Darapladib and Cyclen, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[069] In some embodiments, the method comprises administering to the subject a synergistically effective amount of a pharmaceutical composition comprising Darapladib and Cyclen, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[070] In some embodiments, the method comprises administering to the subject a pharmaceutical composition comprising Darapladib and Cyclen in a molar per molar ratio ranging froml00:l to 1:100, 50:1 to 1:50, 30:1 to 1:30, 15:1 to 1:15, 10:1 to 1:10, 8:1 to 1:8, 4:1 to 1:4, 3:1 to 1:3, 2:1 to 1:2, or is 1:1, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[071] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Darapladib and Mebrofenin, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[072] In some embodiments, the method comprises administering to the subject a synergistically effective amount of a pharmaceutical composition comprising Darapladib and Mebrofenin, thereby treating or preventing SARS-CoV-2 virulence in the subject. Each possibility represents a separate embodiment of the invention. embodiments, the method comprises pharmaceutical composition comprising Darapladib and Mebrofenin in a molar per molar ratio ranging from 100:1 to 1:100, 50:1 to 1:50, 30:1 to 1:30, 15:1 to 1:15, 10:1 to 1:10, 8:1 to 1:8, 4:1 to 1:4, 3:1 to 1:3, 2:1 to 1:2, or is 1:1, thereby treating or preventing SARS-CoV- 2 virulence in the subject. Each possibility represents a separate embodiment of the invention.

[074] According to some embodiments, there is provided a method for treating or preventing SARS-CoV-2 virulence in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Flumatinib and Darapladib.

[075] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising Flumatinib and Darapladib in a molar per molar ratio ranging from 100:1 to 1:100, 50:1 to 1:50, 30:1 to 1:30, 15:1 to 1:15, 10:1 to 1:10, 8:1 to 1:8, 4:1 to 1:4, 3:1 to 1:3, 2:1 to 1:2, or is 1:1. Each possibility represents a separate embodiment of the invention.

[076] According to some embodiments, the SARS-CoV-2 E protein channel blocker is Azacytidine, an analog or a salt thereof. According to some embodiments, the SARS-CoV- 2 E protein channel blocker is 5 -Azacytidine.

[077] Azacytidine, as used herein, includes Azacytidine (CAS: 320-67-2; 4-Amino-1-β-D- ribofuranosyl-s-triazin-2(1H)-one), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof. Azacytidine is described, for example in WO2012135405AE The terms “5- Azacytidine” and “Azacytidine” are used herein interchangeably.

[078] According to some embodiments, the invention provides a pharmaceutical composition comprising Memantine, an analog or a salt thereof, for use in the treatment of a viral infection. In some embodiments, the viral infection comprises a coronaviruses infection. In some embodiments, the viral infection comprises an infection by virus having an E protein being an ion channel. In some embodiments, the viral infection is a coronaviruses infection.

[079] According to some embodiments, the SARS-CoV-2 E protein channel blocker is Memantine, an analog or a salt thereof. According to some embodiments, the SARS-CoV-2 E protein channel blocker is memantine hydrochloride.

[080] Memantine, as used herein, includes memantine (CAS: 19982-08-2; 1-amino-3,5- dimethyladamantane), as well as pharmaceutically acceptable salts, solvates, hydrates, or Memantine is described, for example, in 5,891,885, 5,919,826, and 6,187,338.

[081] According to some embodiments, the invention provides a pharmaceutical composition comprising Gliclazide, an analog or a salt thereof, for treating a viral infection. In some embodiments, the viral infection is an infection by virus having an E protein being an ion channel.

[082] According to some embodiments, the SARS-CoV-2 E protein channel blocker is Gliclazide an analog or a salt thereof.

[083] Gliclazide, as used herein, includes gliclazide (CAS: 21187-98-4; 1-(3- azabicyclo(3.3.0)oct-3-yl)-3-(p-tolylsulfonyl)urea) as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof.

[084] According to some embodiments, the SARS-CoV-2 E protein channel blocker is selected from a group including Ginsenoside.

[085] According to some embodiments, the invention provides a pharmaceutical composition comprising Mavorixafor, an analog or a salt thereof, for treating a viral infection.

[086] According to some embodiments, the SARS-CoV-2 E protein channel blocker is Mavorixafor, an analog or a salt thereof. According to some embodiments, the SARS-CoV- 2 E protein channel blocker is Mavorixafor.

[087] Mavorixafor, as used herein, includes Mavorixafor (CAS: 558447-26-0; N-(1H- benzimidazol-2-ylmethyl)-N-[(8S)-5,6,7,8-tetrahydroquinolin-8-yl]butane- 1,4-diamine), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof. Mavorixafor is described, for example, in U.S. Patent US7332605, and as compound 89 from a series of 169 analogues in W02003055876.

[088] According to some embodiments, the invention provides a pharmaceutical composition comprising Saroglitazar, an analog or a salt thereof, for treating a viral infection.

[089] According to some embodiments, the SARS-CoV-2 E protein channel blocker is Saroglitazar, an analog or a salt thereof. According to some embodiments, the SARS-CoV- 2 E protein channel blocker is Saroglitazar Magnesium.

[090] Saroglitazar, as used herein, includes Saroglitazar (CAS: 495399-09-2; (aS)-a- Ethoxy-4-[2-[2-methyl-5-[4-(methylthio)phenyl]-lH-pyrrol-l-yl]ethoxy]benzenepropanoic pharmaceutically acceptable salts, solvates, Saroglitazar is described, for example, in W02016181409.

[091] According to some embodiments, the invention provides a pharmaceutical composition comprising Mebrofenin, an analog or a salt thereof, for treating a viral infection.

[092] According to some embodiments, the SARS-CoV-2 E protein channel blocker is Mebrofenin, an analog or a salt thereof. According to some embodiments, the SARS-CoV- 2 E protein channel blocker is Mebrofenin.

[093] Mebrofenin, as used herein, includes Mebrofenin (CAS: 78266-06-5; 2-[[2-(3- bromo-2,4,6-trimethylanilino)-2-oxoethyl]-(carboxymethyl)amino]acetic acid), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof. Mebrofenin is described, for example, in US9,878,984.

[094] According to some embodiments, the invention provides a pharmaceutical composition comprising Cyclen, an analog or a salt thereof, for treating a viral infection.

[095] According to some embodiments, the SARS-CoV-2 E protein channel blocker is Cyclen, an analog or a salt thereof. According to some embodiments, the SARS-CoV-2 E protein channel blocker is Cyclen.

[096] Cyclen, as used herein, includes Cyclen (CAS: 294-90-6; 1,4,7,10-

Tetraazacyclododecane), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof. Cyclen is described, for example in US9421223B2.

[097] According to some embodiments, the invention provides a pharmaceutical composition comprising Kasugamycin, an analog or a salt thereof, for treating a viral infection.

[098] According to some embodiments, the SARS-CoV-2 E protein channel blocker is Kasugamycin, an analog or a salt thereof. According to some embodiments, the SARS-CoV- 2 E protein channel blocker is Kasugamycin hydrochloride hydrate (CAS: 19408-46-9).

[099] Kasugamycin, as used herein, includes Kasugamycin (CAS: 6980-18-3; 2-amino-2- [(2R,3S,5S,6R)-5-amino-2-methyl-6-[(2R,3S,5S,6S)-2,3,4,5,6- pentahydroxycyclohexyl]oxyoxan-3-yl]iminoacetic acid), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof. Kasugamycin is described, for example in US3358001A.

[0100] According to some embodiments, the invention provides a pharmaceutical composition comprising Plerixafor, an analog or a salt thereof, for treating a viral infection. to some embodiments, the SARS-CoV-2 E Plerixafor, an analog or a salt thereof. According to some embodiments, the SARS-CoV-2 E protein channel blocker is Plerixafor octahydrochloride.

[0102] Plerixafor, as used herein, includes Plerixafor (CAS: 155148-31-5; 1-[[4-(1,4,8,11- tetrazacyclotetradec-1-ylmethyl)phenyl]methyl]-1,4,8,l 1-tetrazacyclotetradecane), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof. Plerixafor is described, for example in WO2014125499A1.

[0103] SARS-CoV-2 3a protein, also known as open reading frame 3a (ORF3a), is known to one skilled in the art and has a UniProt Accession no: P0DTC3.

[0104] According to some embodiments, the SARS-CoV-2 3a protein comprises the amino acid sequence as set forth in SEQ ID NO 2: MDLFMRIFTIGTVTLKQGEIKDATPSDFVRATATIPIQASLPFGWLIVGVALLAVFQ S AS KIITFKKRW QF ALS KG VHFVCNFFFFFVT V Y S HFFF V A AGLE APFFYF Y AFV Y FFQS INFVRIIMRFWFC WKCRS KNPFF YD AN YFFC WHTNC YD Y CIP YN S VT S S IVIT S GDGTTS PIS EHD Y QIGG YTEKWES G VKDC V VFHS YFTS D Y Y QF Y S TQFS TDTG VE H VTFFIYNKIVDEPEEH V QIHTIDGS S G VVNP VMEPIYDEPTTTT S VPF According to some embodiments, the SARS-CoV-2 3a protein_comprises an analog of SEQ ID NO: 2, such as an analog having at least 85%, at least 90%, at least 95% identity to SEQ ID NO: 2.

[0105] According to some embodiments, the invention provides a pharmaceutical composition comprising Capreomycin, an analog or a salt thereof, for treating a viral infection.

[0106] According to some embodiments, the SARS-CoV-2 3a protein inhibitor is Capreomycin, an analog or a salt thereof. According to some embodiments, the SARS-CoV- 2 3a protein inhibitor is Capreomycin sulfate.

[0107] Capreomycin, as used herein, includes Capreomycin (CAS: 11003-38-6; IUPAC: (3S)-3,6-diamino-N-[[(2S,5S,8E,11S, 15S)-15-amino-11-[(4R)-2-amino-3, 4,5,6- tetrahydropyrimidin-4-yl]-8-[(carbamoylamino)methylidene]-2-(hydroxymethyl)- 3,6,9,12,16-pentaoxo-1,4,7,10,13-pentazacyclohexadec-5-yl]methyl]hexanamide; (3S)-3,6- diamino-N-[[(2S,5S,8E,11S,15S)-15-amino-11-[(4R)-2-amino-3,4,5,6- tetrahydropyrimidin-4-yl]-8-[(carbamoylamino)methylidene]-2-methyl-3,6,9,12,16- pentaoxo-1,4,7,10,13-pentazacyclohexadec-5-yl]methyl]hexanamide), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof. to some embodiments, the invention composition comprising Pentamidine, an analog or a salt thereof, for treating a viral infection.

[0109] According to some embodiments, the SARS-CoV-2 E protein channel blocker is Pentamidine, an analog or a salt thereof. According to some embodiments, the SARS-CoV- 23a protein inhibitor is Pentamidine isethionate.

[0110] Pentamidine, as used herein, includes Pentamidine (CAS: 100-33-4; IUPAC: 4,4'- [pentane-l,5-diylbis(oxy)]dibenzenecarboximidamide), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof.

[0111] According to some embodiments, the invention provides a pharmaceutical composition comprising Spectinomycin, an analog or a salt thereof, for treating a viral infection.

[0112] According to some embodiments, the SARS-CoV-2 E protein channel blocker is Spectinomycin, an analog or a salt thereof. According to some embodiments, the SARS- CoV-23a protein inhibitor is Spectinomycin dihydrochloride.

[0113] Spectinomycin, as used herein, includes Spectinomycin (CAS: 1695-77-8; IUPAC: lR,3S,5R,8R,10S,llS,12S,13R,14S)-8,12,14-trihydroxy-5-methyl-ll,13- bis(methylamino)-2,4,9-trioxatricyclo[8.4.0.03,8]tetradecan-7-one), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof.

[0114] According to some embodiments, the invention provides a pharmaceutical composition comprising Kasugamycin, an analog or a salt thereof, for treating a viral infection.

[0115] According to some embodiments, the SARS-CoV-2 3a protein inhibitor is

Kasugamycin, an analog or a salt thereof. According to some embodiments, the SARS-CoV- 23a protein inhibitor is Kasugamycin hydrochloride hydrate.

[0116] Kasugamycin, as used herein, includes Kasugamycin (CAS :6980- 18-3; IUPAC: 2- amino-2-[(2R,3S,5S,6R)-5-amino-2-methyl-6-[(2R,3S,5S,6S)-2,3,4,5,6- pentahydroxycyclohexyl]oxyoxan-3-yl]iminoacetic acid), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof.

[0117] According to some embodiments, the invention provides a pharmaceutical composition comprising Plerixafor, an analog or a salt thereof, for treating a viral infection. g to some embodiments, the SARS-CoV-23a an analog or a salt thereof. According to some embodiments, the SARS-CoV-2 3a protein inhibitor is Plerixafor.

[0119] Plerixafor, as used herein, includes Plerixafor (CAS: 155148-31-5; IUPAC: 1,1’- (1,4-phenylenebismethylene)bis(1,4,8,ll- tetraazacyclotetradecane)), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof.

[0120] According to some embodiments, the invention provides a pharmaceutical composition comprising Flumatinib, an analog or a salt thereof, for treating a viral infection.

[0121] According to some embodiments, the SARS-CoV-2 3a protein inhibitor is Flumatinib, an analog or a salt thereof. According to some embodiments, the SARS-CoV-2 3a protein inhibitor is Flumatinib.

[0122] Flumatinib, as used herein, includes Flumatinib (CAS: 895519-90-1; IUPAC: 4-[(4- methylpiperazin-l-yl)methyl]-N-[6-methyl-5-[(4-pyridin-3-ylpyrimidin-2- yl)amino]pyridin-3-yl]-3-(trifluoromethyl)benzamide), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof.

[0123] According to some embodiments, the invention provides a pharmaceutical composition comprising Litronesib, an analog or a salt thereof, for treating a viral infection.

[0124] According to some embodiments, the SARS-CoV-2 3a protein inhibitor is

Litronesib, an analog or a salt thereof. According to some embodiments, the SARS-CoV-2 3a protein inhibitor is Litronesib.

[0125] Litronesib, as used herein, includes Litronesib (CAS: 910634-41-2; IUPAC: N- [(5R)-4-(2,2-dimethylpropanoyl)-5-[[2-(ethylamino)ethylsulfonylamino]methyl]-5-phenyl- 1,3,4-thiadiazol-2-yl]-2,2-dimethylpropanamide), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof.

[0126] According to some embodiments, the invention provides a pharmaceutical composition comprising Darapladib, an analog or a salt thereof, for treating a viral infection.

[0127] According to some embodiments, the SARS-CoV-2 3a protein inhibitor is

Darapladib, an analog or a salt thereof. According to some embodiments, the SARS-CoV-2 3a protein inhibitor is Darapladib.

[0128] Darapladib, as used herein, includes Darapladib (CAS: 356057-34-6; IUPAC: N-(2- Diethylaminoethyl)-2-[2-[(4-fluorophenyl)methylsulfanyl]-4-oxo-6,7-dihydro-5H-

as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof.

[0129] According to some embodiments, the invention provides a pharmaceutical composition comprising Floxuridine, an analog or a salt thereof, for treating a viral infection.

[0130] According to some embodiments, the SARS-CoV-2 3a protein inhibitor is

Floxuridine, an analog or a salt thereof. According to some embodiments, the SARS-CoV- 2 3a protein inhibitor is Floxuridine.

[0131] Floxuridine, as used herein, includes Floxuridine (CAS: 50-91-9; IUPAC: 5-Fluoro- 1-[4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-lH-pyrimidine-2,4-dione), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof.

[0132] According to some embodiments, the invention provides a pharmaceutical composition comprising Fludarabine, an analog or a salt thereof, for treating a viral infection.

[0133] According to some embodiments, the SARS-CoV-2 3a protein inhibitor is

Fludarabine, an analog or a salt thereof. According to some embodiments, the SARS-CoV- 2 3a protein inhibitor is Fludarabine.

[0134] Fludarabine, as used herein, includes Fludarabine (CAS: 21679-14-1; IUPAC: [(2R,3S,4S,5R)-5-(6-amino-2-fluoro-purin-9-yl)- 3,4-dihydroxy-oxolan-2- yl]methoxyphosphonic acid), as well as pharmaceutically acceptable salts, solvates, hydrates, or mixtures thereof.

[0135] According to some embodiments, the subject is a subject infected or suspected as being infected by SARS-CoV-2. In some embodiments, the subject is a human subject.

[0136] According to some embodiments, the invention provides at least two SARS-CoV-2 3a protein inhibitors for use in the treatment or prevention of SARS-CoV-2 virulence, in a subject in need thereof.

[0137] According to some embodiments, the invention provides at least two SARS-CoV-2 3a protein inhibitors for use in the prevention of SARS-CoV-2 release from a cell.

[0138] According to some embodiments, the at least two SARS-CoV-23a protein inhibitors are within a pharmaceutical composition.

[0139] In some embodiments, the viral infection is an infection by virus having a 3a protein.

[0140] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Mavorixafor in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Flumatinib. to some embodiments, there is provided a enhancing the anti-viral activity of Cyclen in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Flumatinib.

[0142] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Mebrofenin in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Flumatinib.

[0143] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Saroglitazar in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Flumatinib.

[0144] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Plerixafor in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Flumatinib.

[0145] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Gliclazide in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Flumatinib.

[0146] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Flumatinib in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Saroglitazar.

[0147] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Flumatinib in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Plerixafor.

[0148] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Flumatinib in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Gliclazide.

[0149] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Flumatinib in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Mavorixafor.

[0150] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Flumatinib in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Cyclen. to some embodiments, there is provided enhancing the anti-viral activity of Flumatinib in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Mebrofenin.

[0152] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Mavorixafor in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Darapladib.

[0153] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Cyclen in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Darapladib.

[0154] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Mebrofenin in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Darapladib.

[0155] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Darapladib in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Mavorixafor.

[0156] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Darapladib in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Cyclen.

[0157] According to some embodiments, there is provided a method for increasing or enhancing the anti-viral activity of Darapladib in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Mebrofenin.

[0158] According to another aspect, there is provided a method for increasing or enhancing the anti- viral activity of Flumatinib in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Darapladib.

[0159] According to another aspect, there is provided a method for increasing or enhancing the anti-viral activity of Darapladib in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising Flumatinib.

[0160] In some embodiments, increase, increasing, enhance, or enhancing is at least 5%, 10%, 35%, 50%, 80%, 100%, 150%, 270%, 400%, 650%, 800%, or 1,000% increase compared to a control, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. herein, the term “anti-viral activity” encompasses hampering, inhibiting, any equivalent thereof, or any combination thereof, of viral: genome replication, mRNA synthesis, budding, internalization, entry to a cell, uncoating, release from a cell, or any combination thereof.

[0162] In some embodiments, the cell is a host cell.

[0163] In some embodiments, the host is a mammal host. In some embodiments, the host is a human host or a subject, as disclosed herein.

[0164] As used herein, the terms “treatment” or “treating” of a disease, disorder, or condition encompasses alleviation of at least one symptom thereof, a reduction in the severity thereof, or inhibition of the progression thereof. Treatment need not mean that the disease, disorder, or condition is totally cured. To be an effective treatment, a useful composition herein needs only to reduce the severity of a disease, disorder, or condition, reduce the severity of symptoms associated therewith, or provide improvement to a patient or subject’s quality of life.

[0165] As used herein, the term “prevention” of a disease, disorder, or condition encompasses the delay, prevention, suppression, or inhibition of the onset of a disease, disorder, or condition. As used in accordance with the presently described subject matter, the term "prevention" relates to a process of prophylaxis in which a subject is exposed to the presently described compositions or formulations prior to the induction or onset of the disease/disorder process. The term "suppression" is used to describe a condition wherein the disease/disorder process has already begun but obvious symptoms of the condition have yet to be realized. Thus, the cells of an individual may have the disease/disorder, but no outside signs of the disease/disorder have yet been clinically recognized. In either case, the term prophylaxis can be applied to encompass both prevention and suppression. Conversely, the term "treatment" refers to the clinical application of active agents to combat an already existing condition whose clinical presentation has already been realized in a patient.

[0166] In some embodiments, preventing comprises reducing the disease severity, delaying the disease onset, reducing the disease cumulative incidence, or any combination thereof.

[0167] As used herein, the terms “administering,” “administration,” and like terms refer to any method which, in sound medical practice, delivers a composition containing an active agent to a subject in such a manner as to provide a therapeutic effect. herein, the terms “subject” or “individual” or “mammal,” refers to any subject, particularly a mammalian subject, for whom therapy is desired, for example, a human.

[0169] In some embodiments, a therapeutically effective dose of the composition of the invention is administered. The term "therapeutically effective amount" refers to an amount of a drug effective to treat a disease or disorder in a mammal. The term “a therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result. The exact dosage form and regimen would be determined by the physician according to the patient's condition.

[0170] The dosage administered will be dependent upon the age, health, and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment, and the nature of the effect desired. The route of administration of the pharmaceutical compositions will depend on the disease or condition to be treated. Suitable routes of administration include, but are not limited to, parenteral injections, e.g., intradermal, intravenous, intramuscular, intralesional, subcutaneous, intrathecal, and any other mode of injection as known in the art. Although the bioavailability of peptides administered by other routes can be lower than when administered via parenteral injection, by using appropriate compositions it is envisaged that it will be possible to administer the compositions of the invention via transdermal, oral, rectal, vaginal, topical, nasal, inhalation and ocular modes of treatment. In addition, it may be desirable to introduce the pharmaceutical compositions of the invention by any suitable route, including intraventricular and intrathecal injection; intraventricular injection may be facilitated by an intraventricular catheter, for example, attached to a reservoir.

Pharmaceutical compositions

[0171] According to some embodiments, there is provided a pharmaceutical composition comprising a SARS-CoV-2 3a protein and a SARS-CoV-2 E channel blocker. Each possibility represents a separate embodiment of the invention.

[0172] In some embodiments, the pharmaceutical composition comprises a SARS-CoV-2 E channel blocker being selected from: Mavorixafor, Cyclen, or Mebrofenin, or any combination thereof.

[0173] According to some embodiments, there is provided a pharmaceutical composition comprising Flumatinib and Darapladib.

[0174] In some embodiments, the pharmaceutical composition is for use in treatment or prevention of SARS-CoV-2 virulence in a subject in need thereof. to some embodiments, there is provided a 2 3a protein inhibitor and a SARS-CoV-2 E channel blocker, for use in the treatment or prevention of SARS-CoV-2 virulence in a subject in need thereof.

[0176] In some embodiments, according to the herein disclosed combination, the SARS CoV-2 3a protein inhibitor is formulated within a first pharmaceutical composition and the SARS-CoV-2 E channel blocker, as disclosed herein, is formulated within a second pharmaceutical composition.

[0177] According to some embodiments, there is provided a combination of Flumatinib and Darapladib, for use in the treatment or prevention of SARS-CoV-2 virulence in a subject in need thereof.

[0178] In some embodiments, prevention as disclosed herein for the pharmaceutical composition of the invention or the combination of the invention, comprises prevention of: SARS-CoV-2 entry to a cell of a subject, uncoating of a SARS-CoV-2, release of A SARS- CoV-2 from a cell of a subject, or any combination thereof.

[0179] In some embodiments, the composition of the invention is delivered orally. In some embodiments, the composition of the invention is an oral composition. In some embodiments, the composition of the invention further comprises orally acceptable carrier, excipient, or a diluent.

[0180] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, adjuvant or excipient.

[0181] As used herein, the terms “carrier”, “adjuvant”, or “excipient” refer to any component of a pharmaceutical composition that is not the active agent. As used herein, the term “pharmaceutically acceptable carrier” refers to non-toxic, inert solid, semi-solid liquid filler, diluent, encapsulating material, formulation auxiliary of any type, or simply a sterile aqueous medium, such as saline. Some examples of the materials that can serve as pharmaceutically acceptable carriers are sugars, such as lactose, glucose and sucrose, starches such as corn starch and potato starch, cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt, gelatin, talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as propylene glycol, polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate, agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline, Ringer's solution; and phosphate buffer solutions, as well as other substances used in pharmaceutical formulations. Some non-limiting examples of substances which can serve as a carrier herein include sugar, starch, cellulose and its derivatives, powered tragacanth, malt, gelatin, talc, stearic acid, magnesium stearate, calcium sulfate, vegetable oils, polyols, alginic acid, pyrogen-free water, isotonic saline, phosphate buffer solutions, cocoa butter (suppository base), emulsifier as well as other non-toxic pharmaceutically compatible substances used in other pharmaceutical formulations. Wetting agents and lubricants such as sodium lauryl sulfate, as well as coloring agents, flavoring agents, excipients, stabilizers, antioxidants, and preservatives may also be present. Any nontoxic, inert, and effective carrier may be used to formulate the compositions contemplated herein. Suitable pharmaceutically acceptable carriers, excipients, and diluents in this regard are well known to those of skill in the art, such as those described in The Merck Index, Thirteenth Edition, Budavari et al., Eds., Merck & Co., Inc., Rahway, N.J. (2001); the CTFA (Cosmetic, Toiletry, and Fragrance Association) International Cosmetic Ingredient Dictionary and Handbook, Tenth Edition (2004); and the “Inactive Ingredient Guide,” U.S. Food and Drug Administration (FDA) Center for Drug Evaluation and Research (CDER) Office of Management, the contents of all of which are hereby incorporated by reference in their entirety. Examples of pharmaceutically acceptable excipients, carriers and diluents useful in the present compositions include distilled water, physiological saline, Ringer's solution, dextrose solution, Hank's solution, and DMSO. These additional inactive components, as well as effective formulations and administration procedures, are well known in the art and are described in standard textbooks, such as Goodman and Gillman’s: The Pharmacological Bases of Therapeutics, 8th Ed., Gilman et al. Eds. Pergamon Press (1990); Remington’s Pharmaceutical Sciences, 18th Ed., Mack Publishing Co., Easton, Pa. (1990); and Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott Williams & Wilkins, Philadelphia, Pa., (2005), each of which is incorporated by reference herein in its entirety. The presently described composition may also be contained in artificially created structures such as liposomes, ISCOMS, slow-releasing particles, and other vehicles which increase the half-life of the peptides or polypeptides in serum. Liposomes include emulsions, foams, micelles, insoluble monolayers, liquid crystals, phospholipid dispersions, lamellar layers and the like. Liposomes for use with the presently described peptides are formed from standard vesicle-forming lipids which generally include neutral and negatively charged phospholipids and a sterol, such as cholesterol. The selection of lipids is generally determined by considerations such as liposome size and stability in the of methods are available for preparing liposomes a by Coligan, J. E. et al, Current Protocols in Protein Science, 1999, John Wiley & Sons, Inc., New York, and see also U.S. Pat. Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369.

[0182] The carrier may comprise, in total, from about 0.1% to about 99.99999% by weight of the pharmaceutical compositions presented herein.

General

[0183] As used herein, the term "about" when combined with a value refers to plus and minus 10% of the reference value. For example, a length of about 1,000 nanometers (nm) refers to a length of 1,000 nm ± 100 nm.

[0184] It is noted that as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a polynucleotide" includes a plurality of such polynucleotides and reference to "the polypeptide" includes reference to one or more polypeptides and equivalents thereof known to those skilled in the art, and so forth. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as "solely," "only" and the like in connection with the recitation of claim elements or use of a "negative" limitation.

[0185] In those instances where a convention analogous to "at least one of A, B, and C, etc." is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., "a system having at least one of A, B, and C" would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and/or phrase presenting two or more alternative terms, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase "A or B" will be understood to include the possibilities of "A" or "B" or "A and B."

[0186] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination. All combinations of the embodiments pertaining to the invention are specifically embraced by the present invention and are disclosed herein just as if each and was individually and explicitly disclosed. combinations of the various embodiments and elements thereof are also specifically embraced by the present invention and are disclosed herein just as if each and every such sub-combination was individually and explicitly disclosed herein.

[0187] Additional objects, advantages, and novel features of the present invention will become apparent to one ordinarily skilled in the art upon examination of the following examples, which are not intended to be limiting. Additionally, each of the various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below finds experimental support in the following examples.

[0188] Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples.

EXAMPLES

[0189] Generally, the nomenclature used herein, and the laboratory procedures utilized in the present invention include molecular, biochemical, microbiological and recombinant DNA techniques. Such techniques are thoroughly explained in the literature. See, for example, "Molecular Cloning: A laboratory Manual" Sambrook et al., (1989); "Current Protocols in Molecular Biology" Volumes I-III Ausubel, R. M., ed. (1994); Ausubel et al., "Current Protocols in Molecular Biology", John Wiley and Sons, Baltimore, Maryland (1989); Perbal, "A Practical Guide to Molecular Cloning", John Wiley & Sons, New York (1988); Watson et al., "Recombinant DNA", Scientific American Books, New York; Birren et al. (eds) "Genome Analysis: A Laboratory Manual Series", Vols. 1-4, Cold Spring Harbor Laboratory Press, New York (1998); methodologies as set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659 and 5,272,057; "Cell Biology: A Laboratory Handbook", Volumes I-III Cellis, J. E., ed. (1994); "Culture of Animal Cells - A Manual of Basic Technique" by Freshney, Wiley-Liss, N. Y. (1994), Third Edition; "Current Protocols in Immunology" Volumes I-III Coligan J. E., ed. (1994); Stites et al. (eds), "Basic and Clinical Immunology" (8th Edition), Appleton & Lange, Norwalk, CT (1994); Mishell and Shiigi (eds), "Strategies for Protein Purification and Characterization - A Laboratory Course Manual" CSHL Press (1996); all of which are incorporated by reference. Other general references are provided throughout this document.

Materials and Methods of Vero E6 cells in the presence of SARS-CoV-2 grown in 96-well plates with a culture density of 10,000 cells per well. The culture media contained 10% FCS. After 24 hours, the culture media was replaced to media with 2% FCS and appropriate drugs with a final DMSO concentration of 0.1%. After one hour the cells were infected with virus at an multiplicity of infection (MOI) of 0.01. After 48 hours cell viability was monitored by MTS according to standard protocol from the manufacturer (Promega, USA).

[0191] Results were normalized relative to uninfected cells. Further comparisons may be gained by comparing the data to untreated cells and to cells that received only 0.1% DMSO. All infection experiments were performed in a BSF-3 facility.

Screen in bacteria

[0192] Two channels were targeted for inhibition: 3a and E.

[0193] Using three bacteria-based assays, the inventors screened a library of 2,839 approved for human use drugs and identified ten inhibitors against 3a and eight against E (Table 1).

[0194] The screen was at high concentration (100 μM)

Table 1. E protein and 3a protein inhibitors

EXAMPLE 1

Activity of sole compounds in a “whole virus” system

[0195] The inventors examined the activity of each of the chemicals in vitro by testing their ability to protect cells against viral death.

[0196] Cell were infected by the virus (1:100) and after 48 hours their viability was tested. [0197] Without treatment roughly 50% of the cells died.

[0198] Results were compared relative to uninfected cells (100%), and to cells that received a drug “placebo” (0.1% DMSO).

[0199] Most compounds exhibited activity at varying extents. E inhibitors were found to have little/reduced activity at concentrations of 3 μM or lower (data not shown).

[0200] The two most effective compounds on their own were Darapladib and Flumatinib (which inhibit 3 a).

[0201] Specifically, the results show that in the presence of Flumatinib at concentrations of 0.1 to 3 μM, approximately 60% or more cells have survived. In particular, Flumatinib at 3 μM, provided near full survival (Fig. 1). Further, in the presence of Darapladib at concentrations of 1 to 3 μM, approximately 60% or more cells have survived. In particular, Darapladib at 3 μM, provided full survival (Fig. 1).

EXAMPLE 2

Synergistic effects of E protein and 3a protein inhibitors

[0202] The inventors further sought to examin whether a combination of E inhibitors and 3 a inhibitors would provide a better protection against viral-induced cell death.

[0203] The inventors tested different combinations of Flumatinib (which inhibit 3 a) with various E inhibitors (Mavorixafor, Cyclen, and Sargolitazar) at various combinations (0.1 μM - 3 μM). The inventors showed particularily that Flumatinib and either Mavorixafor or Cyclen at a molar per molar ratio ranging from 3:1 to 1:3 results in a synergistic positive effect on cells survival, providing more than 80% viability (Figs. 2A-2B). Specifically, Flumatinib and Mavorixafor at concentrations of either 0.1 μM and 0.3 μM or 0.3 μM and 0.1 μM provided the synergitic and increased cell viability effect (Fig. 2A). Further, Flumatinib and Cyclen at concentrations of 1 μM and 1 μM provided a synergitic effect, resulting with more than 90% cell viability (Fig. 2B). the inventors have shown that at a signet E inhibitors positively reacted to a combination with Flumatinib (Fig. 4B). Further, the inventors showed that Darapladib exhibited a synergistic effect on the cell survival when provided along with Mavorixafor, Cyclen, and Plerixafor (Fig. 4C).

EXAMPLE 3

Synergistic effects of 3a protein inhibitors

[0205] The inventors further sought to examine whether a particular combination of 3a inhibitors would provide a better protection against viral-induced cell death.

[0206] The inventors tested different combinations of Flumatinib and Darapladib at various combinations (0.1 μM - 3 μM).

[0207] The inventors showed that Flumatinib and Darapladib at a molar per molar ratio ranging from 10:1 to 1:10 results in a synergistic positive effect on cells survival, providing approximately 80% viability, or more (Figs. 3A-3C). Specifically, in the presence of Flumatinib and Darapladib at concentrations of either 1 μM and 0.3 μM or 0.1 μM and 1 μM, respectively, approximately 80% of the cells survived (Figs. 3A-3B). Further, Flumatinib and Darapladib at concentrations of 0.3 μM and 1 μM provided a synergitic effect, resulting with approximately 90% cell viability (Fig. 3C).

[0208] Altough the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.

Claims

What is claimed is:

1. A pharmaceutical composition comprising a SARS-CoV-2 3a protein inhibitor and a SARS-CoV-2 E channel blocker.

2. The pharmaceutical composition of claim 1, wherein said SARS-CoV-2 protein 3a inhibitor is selected from Flumatinib or Darapladib.

3. The pharmaceutical composition of claim 1 or 2, wherein said SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Saroglitazar, Mebrofenin, Cyclen, Plerixafor, and Gliclazide.

4. The pharmaceutical composition of claim 2 or 3, wherein said SARS-CoV-2 protein 3a inhibitor is Flumatinib and said SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Saroglitazar, Mebrofenin, Cyclen, Plerixafor, and Gliclazide.

5. The pharmaceutical composition of claim 2 or 3, wherein said SARS-CoV-2 protein 3a inhibitor is Darapladib and said SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Mebrofenin, and Cyclen.

6. A pharmaceutical composition comprising Flumatinib and Darapladib.

7. The pharmaceutical composition of any one of claims 1 to 6, further comprising a pharmaceutically acceptable carrier.

8. The pharmaceutical composition of any one of claims 1 to 7, for use in treatment or prevention of SARS-CoV-2 virulence in a subject in need thereof.

9. A combination of a SARS-CoV-2 3a protein inhibitor and a SARS-CoV-2 E channel blocker, for use in the treatment or prevention of SARS-CoV-2 virulence in a subject in need thereof, wherein said SARS-CoV-23a protein inhibitor and a SARS-CoV-2 E channel blocker are provided at a molar per molar ratio ranging from 10:1 to 1 : 10 to said subject.

10. The combination of claim 9, wherein said SARS-CoV-2 3a protein inhibitor is formulated within a first pharmaceutical composition and said SARS-CoV-2 E channel blocker is formulated within a second pharmaceutical composition. selected from Flumatinib or Darapladib.

12. The combination of any one of claims 9 to 11, wherein said SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Saroglitazar, Mebrofenin, Cyclen, Plerixafor, and Gliclazide.

13. The combination of claim 11 or 12, wherein said SARS-CoV-2 protein 3a inhibitor is Flumatinib and said SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Saroglitazar, Mebrofenin, Cyclen, Plerixafor, and Gliclazide.

14. The combination of claim 11 or 12, wherein said SARS-CoV-2 protein 3a inhibitor is Darapladib and said SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Mebrofenin, and Cyclen.

15. A combination of Flumatinib and Darapladib, for use in the treatment or prevention of SARS-CoV-2 virulence in a subject in need thereof.

16. The pharmaceutical composition of any one of claims 1 to 8 or the combination of any one of claims 9 to 15, wherein said prevention comprises prevention of any one of: SARS- CoV-2 entry to a cell of said subject, uncoating of said SARS-CoV-2, release of said SARS- CoV-2 from a cell of said subject, or any combination thereof.

17. A method for treating or preventing SARS-CoV-2 virulence in a subject in need thereof, the method comprising administering to said subject a therapeutically effective amount of a pharmaceutical composition comprising a SARS-CoV-2 3a protein inhibitor and a SARS-CoV-2 E channel blocker, thereby treating or preventing SARS-CoV-2 virulence in said subject.

18. The method of claim 17, wherein said SARS-CoV-2 protein 3a inhibitor is selected from Flumatinib or Darapladib.

19. The method of claim 17 or 18, wherein said SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Saroglitazar, Mebrofenin, Cyclen, Plerixafor, and Gliclazide. Flumatinib and said SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Saroglitazar, Mebrofenin, Cyclen, Plerixafor, and Gliclazide.

21. The method of claim 18 or 19, wherein said SARS-CoV-2 protein 3a inhibitor is Darapladib and said SARS-CoV-2 E channel blocker is selected from the group consisting of: Mavorixafor, Mebrofenin, and Cyclen.

22. A method for treating or preventing SARS-CoV-2 virulence in a subject in need thereof, the method comprising administering to said subject a therapeutically effective amount of a pharmaceutical composition comprising Flumatinib and Darapladib.

23. The method of any one of claims 17 to 22, wherein said preventing comprises preventing any one of: SARS-CoV-2 entry to a cell of said subject, uncoating of said SARS- CoV-2 in a cell of said subject, release of said SARS-CoV-2 from a cell of said subject, and any combination thereof.

24. The method of any one of claims 17 to 23, wherein said subject is infected or suspected of being infected by SARS-CoV-2.