[go: up one dir, main page]

WO2015164956A1 - Composés dérivés de benzisothiazole utilisés comme agents thérapeutiques et leurs méthodes d'utilisation - Google Patents

Composés dérivés de benzisothiazole utilisés comme agents thérapeutiques et leurs méthodes d'utilisation Download PDF

Info

Publication number
WO2015164956A1
WO2015164956A1 PCT/CA2015/000291 CA2015000291W WO2015164956A1 WO 2015164956 A1 WO2015164956 A1 WO 2015164956A1 CA 2015000291 W CA2015000291 W CA 2015000291W WO 2015164956 A1 WO2015164956 A1 WO 2015164956A1
Authority
WO
WIPO (PCT)
Prior art keywords
hiv
ome
conhoh
nhoh
compound
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CA2015/000291
Other languages
English (en)
Inventor
David Scott Grierson
Peter K. Cheung
Benoit Chabot
P. Richard Harrigan
Alan Walter COCHRANE
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of British Columbia
University of Toronto
SOCPRA Sciences Sante et Humaines sec
Original Assignee
University of British Columbia
University of Toronto
SOCPRA Sciences Sante et Humaines sec
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of British Columbia, University of Toronto, SOCPRA Sciences Sante et Humaines sec filed Critical University of British Columbia
Publication of WO2015164956A1 publication Critical patent/WO2015164956A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • This invention relates to therapeutics, their uses and methods for the treatment of viral infection, cancer treatment or neurodegenerative disease treatment.
  • the invention relates to therapies and methods of treatment for HIV infection, adenovirus infection, colon cancer and spinal muscular atrophy.
  • HIV Human Immunodeficiency Virus
  • AIDS Acquired Immunodeficiency Syndrome
  • CD4+T cells helper T cells
  • HIV is treated as a chronic infection, rather than a life threatening illness.
  • HIV In common with other viruses, HIV cannot ensure its own replication. Therefore, upon entry into the host cell it must take control of the cell machinery in order to replicate.
  • the virus first attaches to a host cell receptor CD4, then interacts with CCR5 and/or CXCR4 co- receptors, and this is followed by virus-cell fusion.
  • the viral capsid is released into the cytoplasm of the host cell where it is uncoated, releasing the viral enzymes and viral RNA.
  • the single stranded viral RNA is then converted to a double stranded complimentary viral DNA (cDNA) by the viral enzyme RT.
  • the cDNA is translocated to the nucleus where it is incorporated into the host DNA, this step is facilitated by the viral enzyme integrase.
  • the integrated viral DNA must be first transcribed into mRNA in the nucleus and then translated into the viral protein Gag-Pol, in the cytoplasm.
  • Gag-Pol is then cleaved by the viral enzyme protease into gag, gag-pol and env. Further processing by protease yields the remaining essential, structural HIV-i proteins.
  • the viral proteins then assemble at the cell membrane and bud off as a new immature virus.
  • RT The three key enzymes, RT, integrase and protease, in the HIV replication cycle are the major targets against which most anti-HIV/AIDS drugs have been developed.
  • the 9 kb HIV primary transcript is spliced and alternatively spliced through the use of four 5' splice sites (Di to D4) and eight 3' splice sites (Ai, A2, A3, A4a, ⁇ - , A4c, A5 andAz) to produce 15 viral proteins.
  • auxiliary proteins such as Tat, Rev and Nef, will respectively stimulate HIV transcription, allow transport to the cytoplasm of unspliced and partially spliced transcripts encoding viral enzymes and structural proteins, and ensure full virulence in the host (Li, L, H. S. Li, et al. (2005).
  • hnRNP and SR proteins also regulate the alternative splicing of cellular pre-mRNAs (Bakkour, N., Y. L. Lin, et al. (2007). "Small-molecule inhibition of HIV pre-mRNA splicing as a novel antiretroviral therapy to overcome drug resistance.”
  • the fact that HIV produces over 40 distinct mRNAs by alternative splicing may render its replication extraordinarly sensitive to even slight disturbances in the production of early HIV proteins.
  • SR proteins The localization and activity of these proteins are controlled by a variety of kinases such as SRPKs and CLKs (Zhou, Z. and X. D. Fu (2013). "Regulation of splicing by SR proteins and SR protein-specific kinases.” Chromosoma 122(3): 191-207).
  • a small molecule inhibitor of CLKs can alter HIV RNA processing and inhibit HIV replication (Wong, R., A. Balachandran, et al. (2011). "Differential effect of CLK SR Kinases on HIV-i gene expression: potential novel targets for therapy.” Retrovirology 8: 47).
  • This invention is based in part on the fortuitous discovery that compounds described herein modulate HIV infection. Specifically, compounds identified herein, show a reduction of HIV infection of cells, adenovirus infection, colon cancer (cancer) and spinal muscular atrophy (neurodegenerative diseases).
  • J i may be selected from OH; OMe; and
  • J 2 may be selected from OMe; ;
  • NH HO ⁇ J 3 may be selected from OH; OMe; ; an d
  • J 4 may be selected from OH; OMe; ;
  • M t may be selected from H, alkyl (1-6 carbon); F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR; CONR2; CONHOH; Ar; O ⁇ ->MMee - ; // - ; ; ⁇ and
  • M 2 maybe selected from H, alkyl (1-6 carbon); F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR; and
  • M 3 may be selected from H, alkyl (1-6 carbon); F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; NR; CONRa; CONHOH; Ar; OMe ; / ; « ; and
  • M 6 may be selected from H, alkyl (1-6 carbon); F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; NR; C0NR 2 ; CONHOH; Ar; OMe ; / ; ⁇ ; and
  • M 7 may be selected from H, alkyl (1-6 carbon); F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; NR; ; CONR2; CONHOH; Ar; OMe ; / ; ; an d
  • M 8 maybe selected from H, alkyl (1-6 carbon); F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; NR 2 ; C0NR 2 ; CONHOH; Ar; OMe ; / ; f ; an d
  • M 9 may be selected from H, alkyl (1-6 carbon); F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; NR; CONR2; CONHOH; Ar; OMe ; / ; f ; an d
  • R may be selected from H; Me; Et; Pr; Bu; CF 3 ; and NHOH;
  • L may be NH
  • Qi may be selected from N0 2 ; CN; CF 3 ; and NH 2 ;
  • Q 2 may be selected from N0 2 ; CN: CF 3 ; and NH 2 ;
  • Q 3 maybe selected from N0 2 ; CN; CF 3 ; and NH 2 ;
  • Q 4 maybe H; or Me; Et; H, alkyl (1-6 carbon); F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR A ; A 2 ;C0NR A 2 ;C0NH0H;Ar; OMe ; /" ; u ⁇ ⁇ ; and
  • Q 7 may be selected from H; Me; Et; H, alkyl (1-6 carbon); F; CI; Br; I; COH; CF 3 ; CN;
  • Qs may be selected from H; Me; Et; H, alkyl (1-6 carbon); F; CI; Br; I; COH; CF 3 ; CN;
  • R A may be selected from H; Me; Et; Pr; Bu; CF 3 ; and NHOH.
  • J 3 may be selected from OMe; and >0 ⁇ ;
  • J 4 may be selected from OMe; and ° e ;
  • Mi may be selected from H, Me; OMe - / ⁇ f ; and
  • M 2 may be selected from H, Me;
  • R may be selected from H; Me; Et; Pr; Bu; CF 3 ; and COH;
  • L may be NH
  • Qi may be selected from N0 2 ; CN; CF 3 ; and NH 2 ;
  • Q 2 may be selected from N0 2 ; CN; CF 3 ; and NH 2 ;
  • Q 3 may be selected from N0 2 ; CN; CF 3 ; and NH 2 ;
  • Q 4 may be H; or Me.
  • Formula wwhheerreeiinn ma be sel d from one of the
  • Jx may be selected from OH; OMe and OMe
  • J 2 may be selected from OMe;
  • J 3 may be selected from OMe;
  • J 4 may be selected from OMe;
  • Mi may be selected from Me
  • M 2 may be selected from Me
  • R may be selected from H; Me; Et; Pr; and Bu;
  • Qi may be selected from N0 2 ;CN; CF 3 ; and NH 2 ;
  • Q 2 may be selected from N0 2 ; CN; CF 3 ; and NH 2 ;
  • Q 3 may be selected from N0 2 ; CN; CF 3 ; and NH 2 .
  • J 2 may be OH or OMe
  • J 3 may be selected from H n M
  • Mi may be selected from Me; °M e ;
  • M 2 may be H or Me
  • R may be selected from H; Me; Et; Pr; and Bu; may be selected from one of the following:
  • Q 2 may be selected from N0 2 ; and NH 2 ;
  • Q 3 may be N0 2 .
  • a pharmaceutical composition including a compound or pharmaceutically acceptable salt described herein and a pharmaceutically acceptable excipient.
  • a method for the treatment of HIV infection, adenovirus infection, colon cancer or spinal muscular atrophy comprising administering to a subject in need thereof a compound or pharmaceutically acceptable salt thereof as described herein.
  • a pharmaceutical composition including a compound or pharmaceutically acceptable salt thereof described herein and a pharmaceutically acceptable excipient.
  • a commercial package including (a) a compound described herein; and (b) instructions for the use thereof for inhibiting HIV infection or adenovirus infection.
  • the instructions may be for the treatment of colon cancer or spinal muscular atrophy.
  • a commercial package including (a) a pharmaceutical composition comprising a compound described herein and a pharmaceutically acceptable carrier; and (b) instructions for the use thereof for inhibiting HIV infection or adenovirus infection.
  • the instructions may be for the treatment of colon cancer or spinal muscular atrophy.
  • the compound may be selected from one or more of the following:
  • the compound may be selected from one or more of the following:
  • ⁇ may be selected from one of the following: ; om one of the following: be selected from one of the following: J2 ; may be selected from one of the following: . may be selected from one of the
  • Ji may be selected from OH; OMe; and .
  • Ji may be selected from OH; and OMe.
  • J i may be may be OH.
  • J x may be OMe.
  • J 2 may be selected from .
  • Ja may be selected from
  • J 2 may be selected fro J 2 maybe selected from OMe; . j 2 ma y be selected from OMe; . J 2 may be selected from OMe; NH 2 HO . J 2 may be selected from OMe; O e ; / and J 2 may be selected from °M e ; / ; and . j 2 may be o
  • J 3 may be selected from OH; OMe; OMe ; / NH 2 H NH 2 ANC [ J 3 may be selected from OH; OMe; °Me; / ; NH 2;an( j
  • an d an d . J 3 may be selected from OH; OMe; OMe ; /" ; an d _ j 3 may J 3 maybe J 3 maybe J 3 may be OH.
  • j 4 may
  • Mi may be selected from H, alkyl (1-6 carbon); F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR; CO;
  • SOaNRs; C0NR 2 ; CONHOH; Ar; O e ; / : ; . M may be selected from H, Me; F; CI; CO; S0 2 NR 2 ; CONR 2 ;
  • Ar; M may be selected from H, Me; Et NR 2 ; C0NR 2 ;
  • Ar; Mi may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR; CO; S0 2 NR 2 ; C0NR 2 ; ⁇ N—
  • M t may be selected from H, Me; Et; ⁇ ; and ⁇ t .
  • Mi may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR; CO; S0 2 NR 2 ;
  • C0NR 2 ; . Mi may be selected from Me; Et; Pr; Bu; F;
  • Mi maybe selected from Me; °M e ; / ;and ⁇ . Mi maybe selected from H; Me;
  • Et; Pr; Bu; OMe ; / an( j f . Mi maybe selected from Me; Et; Pr; Bu;
  • Mi may be selected from
  • OMe Mi may be selected from Me; Et; Pr; COH; CF 3 ; CN; N0 2 ; CO; r .
  • Mi may be selected from H, alkyl (1-6 carbon); F; CI Br I COH CF CN;
  • M 2 maybe selected from H, Me; Et; Pr; Bu; F; CI; Br;
  • S0 2 NR 2 ; C0NR 2 ; CONHOH; Ar; OMe ; / ; M 2 may be selected from H, alkyl (1-6 carbon); F; CI; Br;
  • S0 2 NR 2 ; C0NR 2 ; CONHOH; Ar; OOMMee - ; // - ; M may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR; CO; S0 2 NR 2 ; CONR 2 ; and CONHOH.
  • M 2 may be selected from Me; Et; Pr; Bu; COH; CF 3 ; CN; N0 2 ; NR;
  • M 2 may be selected from H, Me; Et; Pr; Bu; COH; CF 3 ; CN; N0 2 ; NR; CO; OMe ;
  • M 2 may be Me.
  • M 2 may be selected from H, Me; Et; Pr; and Bu.
  • M 2 I COH; CF 3 ; CN; N0 2 ; CO;
  • M 2 may be selected from
  • H, Me; Et; Pr; Bu; F; CI; Br; I; COH; CF M 2 maybe selected from H, Me;
  • M 3 may be selected from H, alkyl -6 carbon); F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; NR; CO;
  • SO.NR,; C0NR 2 ; CONHOH; Ar; . M 3 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; NR; CO; S0 2 NR 2 ;
  • CONRa; CONHOH; Ar; . M 3 maybe selected from H, Me; Et; Pr; CF 3 ; CN; N0 2 ; CO; CONHOH; Ar; OMe ;
  • M 3 may be selected from H, Me; Et; Pr; Bu; F; CI;
  • M 3 maybe selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • M 3 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; N0 2 ; NR; CO; and CONHOH.
  • M 3 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; and N0 2 .
  • M 3 maybe selected from H, Me; Et; Pr; Bu; F; NHOH; CF 3 ; CN; and N0 2 .
  • M 3 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; and N0 2 .
  • M 3 may be selected from H, Me; Et; Pr; Bu; NHOH; and N0 2 .
  • M 3 may be Me.
  • M 4 maybe selected from H, alkyl (1-6 carbo NR; CO;
  • S0 2 NR 2 ; CONR 2 ; CONHOH; Ar; M 4 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; NR; CO; S0 2 NR 2 ;
  • CONR2; CONHOH; Ar; OMe ; / ; M 4 maybe selected from H, Me; Et; Pr; Bu; NHOH; CF 3 ; CN; N0 2 ; CO; CONHOH; Ar; OMe ;
  • M 4 may be selected from H, Me; Et; Pr; Bu; F; CI;
  • M 4 maybe selected from H, Me; Et; Pr; Bu; F; Cl; Br; I; NHOH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • M 4 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; N0 2 ; NR; CO; and CONHOH.
  • M 4 may be selected from H, Me; Et; Pr; Bu; F; Cl; Br; I; NHOH; CF 3 ; CN; and N0 2 .
  • M 4 maybe selected from H, Me; Et; Pr; Bu; F; NHOH; CF 3 ; CN; and N0 2 .
  • M 4 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; and N0 2 .
  • M 4 may be selected from H, Me; Et; Pr; Bu; NHOH; and N0 2 .
  • M 4 may be Me.
  • M 5 may be selected from H, alkyl (1-6 carbon); F; Cl; Br; I; NHOH; CF 3 ; CN; N0 2 ; NR; CO;
  • SO2NR2; C0NR 2 ; CONHOH; Ar; OMe ; / ; M 5 may be selected from H, Me; Et; Pr; Bu; F; Cl; Br; I; NHOH; CF 3 ; NR; CO; S0 2 NR 2 ;
  • C0NR 2 ; CONHOH; Ar; OMe ; / ; . M 5 maybe CF 3 ; CN; N0 2 ; CO; CONHOH; Ar; OMe ;
  • M 5 may be selected from H, Me; Et; Pr; Bu; F; Cl;
  • M 5 maybe selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • M 5 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; N0 2 ; NR; CO; and CONHOH.
  • M 5 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; and N0 2 .
  • M 5 may be selected from H, Me; Et; Pr; Bu; F; NHOH; CF 3 ; CN; and N0 2 .
  • M 5 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; and N0 2 .
  • M 5 may be selected from H, Me; Et; Pr; Bu; NHOH; and N0 2 .
  • M 5 may be Me.
  • M 6 may be selected from H, alkyl (1-6 carbo NR; CO;
  • SOaNRs; CONRa; CONHOH; Ar; OMe ; M 6 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; NR; CO; S0 2 NR 2 ;
  • C0NR 2 ; CONHOH; Ar; OOMMee - ; / ; M 6 maybe selected from H, Me; Et; CF 3 ; CN; N0 2 ; CO; CONHOH; Ar; OMe ; M 6 may be selected from H, Me; Et; Pr; Bu; F; CI;
  • M 6 maybe selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • M 6 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; N0 2 ; NR; CO; and CONHOH.
  • M 6 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; and N0 2 .
  • M 6 may be selected from H, Me; Et; Pr; Bu; F; NHOH; CF 3 ; CN; and N0 2 .
  • M 6 maybe selected from H, Me; Et; Pr; Bu; NHOH; CN; and N0 2 .
  • M 6 may be selected from H, Me; Et; Pr; Bu; NHOH;
  • M 7 may be selected from H, alkyl (1-6 carbon); F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; NR; CO;
  • S0 2 NR 2 ; CONR 2 ; CONHOH; Ar; OOMMee - ; // ⁇ ; M 7 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; NR; CO; S0 2 NR 2 ;
  • M 7 maybe selected from H, Me; Et; Pr; Bu; NHOH; CF 3 ; CN; N0 2 ; CO; CONHOH; Ar; OMe ;
  • M 7 may be selected from H, Me; Et; Pr; Bu; F; CI;
  • M 7 maybe selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • M 7 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; N0 2 ; NR; CO; and CONHOH.
  • M 7 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; and N0 2 .
  • M 7 maybe selected from H, Me; Et; Pr; Bu; F; NHOH; CF 3 ; CN; and N0 2 .
  • M 7 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; and N0 2 .
  • M 7 may be selected from H, Me; Et; Pr; Bu; NHOH; and N0 2 .
  • M 7 may be Me.
  • M 8 may be selected from H, alkyl (1-6 carbo NR; CO;
  • S0 2 NRs>; CON ij CONHOH; Ar; OMe ; M 8 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; NR; CO; S0 2 NR 2 ;
  • C0NR 2 ; CONHOH; Ar; OMe ; / ; M 8 maybe selected from H, Me; Et; CF 3 ; CN; N0 2 ; CO; CONHOH; Ar; OMe ;
  • M 8 maybe selected from H, Me; Et; Pr; Bu; F; CI;
  • M 8 maybe selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • M 8 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; N0 2 ; NR; CO; and CONHOH.
  • M 8 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; and N0 2 .
  • M 8 may be selected from H, Me; Et; Pr; Bu; F; NHOH; CF 3 ; CN; and N0 2 .
  • M 8 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; and N0 2 .
  • M 8 may be selected from H, Me; Et; Pr; Bu; NHOH; and N0 2 .
  • M 8 may be Me.
  • M 9 may be selected from H, alkyl (1-6 carbo NR; CO;
  • S0 2 NR 2 ; C0NR 2 ; CONHOH; Ar; OOMMee - ; M 9 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; NR; CO; S0 2 NR 2 ;
  • M 9 maybe selected from H, Me; Et; Pr; Bu; NHOH; CF 3 ; CN; N0 2 ; CO; CONHOH; Ar; OMe ;
  • M 9 may be selected from H, Me; Et; Pr; Bu; F; CI;
  • M 9 maybe selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • M 9 may be selected from H, Me; Et; Pr; Bu; NHOH; CN; N0 2 ; NR; CO; and CONHOH.
  • M 9 may be selected from H, Me; Et; Pr; Bu; F; CI; Br; I; NHOH; CF 3 ; CN; and N0 2 .
  • M 9 maybe selected from H, Me; Et; Pr; Bu; F; NHOH; CF 3 ; CN; and N0 2 .
  • M 9 maybe selected from H, Me; Et; Pr; Bu; NHOH; CN; and N0 2 .
  • M 9 may be selected from H, Me; Et; Pr; Bu; NHOH; and N0 2 .
  • M 9 may be Me.
  • R may be selected from H; Me; Et; Pr; Bu; CF 3 ; and NHOH.
  • R may be selected from H; Me; Et; Pr; Bu; and NHOH.
  • R may be selected from H; Me; Et; Pr; Bu; and CF 3 .
  • R may be selected from H; Me; Et; Pr; CF 3 ; and NHOH.
  • R may be selected from H; Me; Pr; Bu; CF 3 ; and NHOH.
  • R may be selected from H; Et; Pr; Bu; CF 3 ; and NHOH.
  • R may be selected from H; Et; Pr; Bu; CF 3 ; and NHOH.
  • R may be selected from Me; Et; Pr; Bu; and NHOH.
  • R may be selected from Me; Et; Pr; Bu; and NHOH.
  • R may be selected from H; Me; Et; Pr; and Bu.
  • R may be selected from H; Me; Et; and Pr. R may be selected from H; and Me. R may be selected from H; Me; Et; Pr; and NHOH.
  • R may be H.
  • R may be Me.
  • R may be Et.
  • R may be Pr.
  • R may be Bu.
  • R may be NHOH.
  • L may be NH.
  • ⁇ ⁇ be selected from one of the following:
  • . maybe may be maybe maybe . may be selected from one of the
  • Qi may be selected from N0 2 ; and NH 2 .
  • Qi may be N0 2 .
  • Qi may be NH 2 .
  • Q 2 may be selected from N0 2 ; and NH 2 .
  • Q 2 may be N0 2 .
  • Q 2 may be NH 2 .
  • Q 3 may be selected from N0 2 ; and NH 2 .
  • Q 3 may be N0 2 .
  • Q 3 may be NH 2 .
  • Q 4 may be selected from H; alkyl (1-6 carbon); F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR A ; CO;
  • Q 4 may be selected from H; Me; Et; F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR A ; CO; S0 2 NR A 2 ;
  • C0NR A 2 ; CONHOH; Ar; OMe ; Q 4 may be selected from H; Me ; and .
  • Q 4 may be selected from CI; COH; CF 3 ; CN;
  • Q 4 may be selected from H; Me; Et; F; CI; Br; COH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • Q 4 maybe selected from H; Me; Et; F; CI; Br; COH; CF 3 ; CN; N0 2 ; and CO.
  • Q 4 may be Me.
  • Q 5 may be selected from H; alkyl (1-6 carbon); F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR A ; CO;
  • Q 5 may be selected from H; Me; Et; Pr; Bu; F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; CO; CONHOH;
  • OMe ; / ; Q 5 may be selected from H; Me; Et;
  • F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR A ; CO; S0 2 NR A 2 ; C0NR A 2 ; CONHOH; Ar; OMe ; Q 5 may be selected from H; Me
  • Q 5 may be selected from COH; CF 3 ; CN; N0 2 ; CO; CONHOH; O e ; ; f ; Q 5 may be selected from H; Me; Et; F COH; CF :
  • Q 5 maybe selected from H; Me; Et; F; Cl; Br; COH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • Q 5 may e selected from H; Me; Et; F; Cl; Br; COH; CF 3 ; CN; N0 2 ; and CO.
  • Q 5 may be Me.
  • Q 6 may be selected from H; alkyl (1-6 carbon); F; Cl; Br; I; COH; CF 3 ; CN; N0 2 ; NR A ; CO;
  • Q 6 mayb I; COH; CF 3 ; CN; N0 2 ; CO; CONHOH; Q 6 may be selected from H; Me; Et F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR A ; CO; S0 2 NR A 2 ; CONR A 2 ; CONHOH; Ar; OMe ; ; Et; CF 3 ; CN; N0 2 ; CO; CONHOH; OMe ; / ; ; and
  • Q 6 may be selected from H; Me; Et; F; CI; COH; CF 3 ; CN; N0 2 ; CO; CONHOH; OMe ; ; COH; CF 3 ;
  • Q 6 maybe selected from H; Me; Et; F; CI; Br; COH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • Q 6 may be selected from H; Me; Et; F; CI; Br; COH; CF 3 ; CN; N0 2 ; and CO.
  • Q 6 maybe Me.
  • Q 7 may be selected from H; alkyl (1-6 carbon); F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR A ; CO;
  • Q 7 maybe selected from H; Me; Et; Pr; Bu; F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; CO; CONHOH;
  • OMe- / ⁇ - ; Q 7 may be selected from H; Me; Et;
  • F; CI; Br; I; COH; CF 3 ; CN; N0 2 ; NR A ; CO; S0 2 NR A 2 ; C0NR A 2 ; CONHOH; Ar; OMe ; Q 7 may be selected from H; Me; Et; F; CI; Br;
  • Q 7 may be selected from H; Me; Et; F; CI; COH; CF 3 ; CN; N0 2 ; CO; CONHOH; OMe ; Q 7 may be selected from H; Me; Et; F; COH; CF 3
  • Q 7 maybe selected from H; Me; Et; F; Cl; Br; COH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • Q 7 maybe selected from H; Me; Et; F; Cl; Br; COH; CF 3 ; CN; N0 2 ; and CO.
  • Q 7 may be Me.
  • Q 8 may be selected from H; alkyl (1-6 carbon); F; Cl; Br; I; COH; CF 3 ; CN; N0 2 ; NR A ; CO;
  • Q 8 may be selected from H; Me; Et; Pr; Bu; F; Cl; Br; I; COH; CF 3 ; CN; N0 2 ; CO; CONHOH;
  • 0Me ; / ; Qs may be selected from H; Me; Et;
  • Q 8 may be selected from H; Me; Et; F; Cl; COH; CF 3 ; CN; N0 2 ; CO; CONHOH; OMe ; Q 8 may be selected from H; Me; Et; F; COH;
  • Q 8 may be selected from H; Me; Et; F; Cl; Br; COH; CF 3 ; CN; N0 2 ; CO; and CONHOH.
  • Q 8 may be selected from H; Me; Et; F; Cl; Br; COH; CF 3 ; CN; N0 2 ; and CO.
  • Q 8 maybe Me.
  • R A may be selected from H; Me; Et; Pr; Bu; CF 3 ; and NHOH.
  • R A may be selected from H; Me; Et; Pr; Bu; and CF 3 .
  • R A may be selected from H; Me; Et; Pr; Bu; and NHOH.
  • R A may be selected from H; Me; Et; Pr; CF 3 ; and NHOH.
  • R A may be selected from H; Me; Pr; Bu; CF 3 ; and NHOH.
  • R A may be selected from H; Et; Pr; Bu; CF 3 ; and NHOH.
  • R A may be selected from H; Et; Pr; Bu; CF 3 ; and NHOH.
  • R A may be selected from Me; Et; Pr; Bu; CF 3 ; and NHOH.
  • R A may be selected from Me; Et; Pr; Bu; CF 3 ; and NHOH.
  • R A may be selected from Me; Et; Pr; Bu; CF 3 ; and NHOH
  • R A may be selected from H; Me; Et; and Pr.
  • R A may be selected from H; Me; Et; Pr; and NHOH.
  • R A maybe H.
  • R A may be Me.
  • R A maybe Et.
  • R A maybe Pr.
  • R A maybe Bu.
  • R A may be CFg.
  • R A may be NHOH.
  • R A may be selected from H; Me; Et; CF 3 ; and NHOH.
  • R A may be selected from H; Me; Et; CF 3 ; and NHOH.
  • FIGURE lA shows a graph of the anti-HIV activity of compounds DGPS56/DH-052 (C8),
  • FIGURE lB shows a graph of the anti-HIV activity of compound SL309 at various
  • FIGURE 2 shows a series of plots of the anti-HIV-i activity of C8 against wild-type subtype A and B, and multidrug-resistant clinical isolates (A: HrV-iIIIB, B: 97USSN54 (subtype A), C: (N)NRTI-resistant isolate, D: Pi-resistant isolate E: INI-resistant isolate, F: MVC-resistant R5 strain).
  • FIGURE 3 shows a bar graph summarizing HIV-i inhibition by compounds DGPS56/DH- 052 (C8), DGPS39/LB-45 (E5), DGPS154/MM-99 (D3), and DGPS73/SKK-31 (C2) on HIV-1NL4-3 as compared to negative control DMSO and positive control AZT, where C8 bars are shown by an arrow.
  • FIGURE 4A shows a map of donor and acceptor splice sites used for the production of the major HIV mRNAs (SS - singly spliced; UN - unspliced; MS - multiply spliced).
  • FIGURE 4B shows quantitative RT-PCR assays from HeLa-HIV cells treated with C8.
  • FIGURE 4C shows the intensity of products when normalized relative to the amount
  • FIGURE 5 shows a comparison of the effect of C8 as compared to DMSO on HIV-i
  • FIGURE 6 A shows a map of donor and acceptor splice sites used for the production of the major HIV mRNAs, wherein the AS2 set of primers was used to monitor the production of Tati and Nef2 in panels B and C.
  • FIGURE 6 B shows the results from the endpoint RT-PCR assay from HeLa-HIV cells
  • FIGURE 6C shows the histograms depicting the impact of C8 on Tati and Nef2 production, wherein the intensity of products was normalized relative to the total intensity for each lane.
  • FIGURE 7A shows a bar graph of the relative expression of level of unspliced RNA in the HeLa-HIV cell line with three different primer pairs (US-i, US-2 and US-4) at three different concentrations of C8 (i.e. 1 uM, 5 ⁇ , 10 uM) relative to control.
  • FIGURE 7B shows a bar graph of Tat transactivated HIV promoter activity in response to compound C8 at various concentrations (i.e. 1 ⁇ , ⁇ ⁇ , 2 ⁇ ⁇ ), measured as total GFP fluorescence at 24 hours post-transfection.
  • FIGURE 7C shows a bar graph of the transcription levels of housekeeping cellular genes MRPL19, PUMi and YWHAZ in HeLa-HIV a cell line in response to various concentrations of compound C8.
  • FIGURE 8A HeLa cells were co-transfected with a plasmid carrying Bcl-x minigene X2.13 and a plasmid containing the CMV promoter driving the expression of the SRSFi or SRSF9 cDNA, wherein the assay was performed in the presence of the indicated concentrations of C8 (i.e. 1 ⁇ , 5 ⁇ , io uM) as compared to control.
  • C8 i.e. 1 ⁇ , 5 ⁇ , io uM
  • FIGURE 8B HeLa cells were co-transfected with the Bcl-x minigene X2 and a Flag-SRSFio plasmid in the presence of the indicated concentrations of C8, wherein the percentage of Bcl-S over the sum of both Bcl-xS and Bcl-xL is shown in the histogram.
  • FIGURE 8C shows an immunoblot of the expression of Flag-SRSFio as compared to actin.
  • FIGURE 9A shows a histogram representing quantitative RT-PCR as used to assess the level of depletion of endogenous SRSFio in the HeLa-HIV cell line in the presence of an siRNA targeting SRSFio.
  • FIGURE 9B shows a histogram representing quantitative RT-PCR assays from HeLa-HIV cells treated with siSRSFio for 96 hours, where the histograms show the impact of C8 on selected HIV amplicons, wherein the intensity of products was normalized relative to the amount produced in the controls (which corresponds to an arbitrary unit value of i).
  • FIGURE 10A shows Hela cells expressing Flag-SRSFio or HA-SRSFio were either treated exposed to 20uM of C8 or were left untreated, wherein control cell extracts were treated with calf intestinal phosphatase (CIP) or left untreated (All extracts were separated by protein gel and detected by Western blot).
  • CIP calf intestinal phosphatase
  • FIGURE 10B shows a SRSFio-derived peptide recovered by MALDI mass spectrometry, wherein the phosphorylated serine at position 133 is highlighted by a circle.
  • FIGURE 10C shows cells treated with increasing concentrations of C8 (1, 10, 25 uM) have a reduction in the recovery of the phosphorylated SRSFio-derived peptide by MALDI mass spectrometry as compared to the non-phosphorylated SRSFio-derived peptide.
  • FIGURE 10D shows Hela cells expressing Flag-SRSFio were either exposed to 1 or 10 ⁇ of C8, wherein the extracts were immunoprecipitated with an anti-FLAG antibody and subjected to MALDI mass spectrometry, with treatments having increasing concentrations of C8 resulted in an increased recovery of hTRA2 -specific peptides.
  • FIGURE 10E shows increased recovery of SRPKi and SRPK2 related peptides.
  • FIGURE 11A shows Hela-HIV cells treated with with either DMSO, or various
  • FIGURE nB shows the addition of increasing concentrations of C8 (l ⁇ , ⁇ uM or 20 ⁇ ) altered the shift in splice site use for the BCLAFi gene in three colon cancer cell lines, leading to decreased production of the exon 5a-containing splice variant of BCLAFi.
  • FIGURE 12A shows cell population dot plots of a T-cell CEM-derived GFP reporter cell line (CEM-GXR) used for quantitating toxicity (values were obtained by Guava
  • FIGURE 12B shows the percentage of viable cells in response to increasing concentrations of C8 (i.e. 0.125 uM, 0.25 uM, 0.5 uM, 1 ⁇ , 2 uM, 4 uM, 8 ⁇ and 16 ⁇ ).
  • FIGURE 13B shows cells were treated with varying doses of C8 after infection ( 24 h), when cells were harvested and lysed, then the extracts were then used to determine levels of adenovirus present by limiting dilution titration as shown is a representative line graph.
  • FIGURE 15 shows a graph of SL309 inhibition of viral spread at various concentrations of
  • FIGURE 16 shows the adverse effect of C8 on GXR-CEM cell viability as measured by Guava
  • HIV infection i.e. the prevention and/or treatment of infection by HIV and in the treatment of the resulting acquired immune deficiency syndrome (AIDS)
  • HIV infection is meant to include both the prevention and/or treatment of infection by HIV.
  • the HIV infection may be either latent or acute.
  • a point of attachment bond denotes a bond that is a point of attachment between two chemical entities, one of which is depicted as being attached to the point of attachment bond and the other of which is not depicted as being attached to the point of attachment bond.
  • the specific point of attachment to the non-depicted chemical entity may be specified by inference. For example, the compound CH 3 -R3, wherein " infers that when R3 is "XY", the point of attachment bond is the same bond as the bond by which R3 is depicted as being bonded to CH 3 .
  • latent HIV infection refers to the cellular latency of HIV infection, as contrasted with clinical latency (i.e., the dormancy of HIV in resting memory CD4 T cells and other cells, including but not limited to monocyte/macrophages, neural cells (including glial cells and astrocytes), as well as the cells of the reproductive system). Latency ends when the virus starts to actively replicate within the subject.
  • C x -Cy alkyl is used as it is normally understood to a person of skill in the art and often refers to a chemical entity that has a carbon skeleton or main carbon chain comprising a number from x to y (with all individual integers within the range included, including integers x and y) of carbon atoms.
  • a "d-C 10 alkyl” is a chemical entity that has 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atom(s) in its carbon skeleton or main chain.
  • acute HIV infection refers to the active replication of the HIV within the subject.
  • the treatment may involve the administering to a subject in need of such treatment a pharmaceutical composition comprising a pharmaceutical carrier and a therapeutically-effective amount of a compound described herein.
  • compositions may be in the form of orally-administrable suspensions or tablets; nasal sprays; sterile injectable preparations, for example, as sterile injectable aqueous or oleagenous suspensions or suppositories.
  • compounds of Formula I or Formula II above may be used for systemic treatment of HIV infection.
  • compounds of Formula I or Formula II may be used in the preparation of a medicament or a composition for systemic treatment of HIV infection.
  • methods of systemically treating HIV infection are also provided.
  • compounds described herein may be used for the treatment of adenoviral infections, for the treatment of colon cancer, or spinal muscular dystrophy.
  • compound described herein may be used for the treatment of viral infection.
  • compound described herein may be used for the treatment of neurodegenerative diseases.
  • compound described herein may be used for the treatment of cancer.
  • SRSF10 serine/arginine-rich splicing factor 10
  • SRSF10 serine/arginine-rich splicing factor 10
  • the role of SR proteins in regulated splicing is achieved via binding to exonic splicing enhancer sites that may lead to the inclusion of an exon into the spliced mRNA transcript. Alternatively, SR protein binding may induce skipping of an exon leading to its removal from the spliced mRNA transcript.
  • SR proteins are subjected to a reversible phosphorylation/
  • SRSFio has been shown to be a key regulator of alternative splicing that can function as a sequence-dependent splicing activator (Feng, Y., M. Chen, et al. (2008). "Phosphorylation switches the general splicing repressor SRp38 to a sequence-specific activator.” Nat Struct Mol Biol 15(10): 1040-1048).
  • the SRSFio homolog in mice has been shown to be involved in splicing events related to adipogenesis and adipocyte differentiation, specifically involving the axini, ACLY, UFPi and lipin 1 genes(Ii, H., Y. Cheng, et al. (2014).
  • SRSFio regulates alternative splicing and is required for adipocyte differentiation. Mol Cell Biol 34(12): 2198- 2207).
  • SRSFio was shown to play a regulatory role in variety of splicing events linked to genes involved in cell survival and responses to cellular stress and apoptosis (Zhou, X., W. Wu, et al. (2014). "Transcriptome analysis of alternative splicing events regulated by SRSFio reveals position-dependent splicing modulation.” Nucleic Acids Res 42(6): 4019-4030).
  • SRSFio as a splicing factor involved in the response to heat shock (Shi, Y. and J. L. Manley (2007).
  • SRSFio has been shown to regulate the splicing of Bcl-2-associated transcription factor 1 (BCLAFi) transcript leading to the inclusion of an alternative exonsa.
  • Tissue samples from colorectal cancer patients showed increased SRSFio expression and increased inclusion of the BCLAFi exonsa variant(Zhou, X., X. Li, et al. (2014). "BCLAFi and its splicing regulator SRSFio regulate the tumorigenic potential of colon cancer cells.” Nat Commun 5: 4581).
  • Increased expression of SRSFio has also been shown in human endometrial carcinomas and ovarian cancers by immunohistochemical analysis of tumor tissue (Peng, Z. Y, P. J. Xiao, et al. (2011). "NSSRi is regulated by testosterone in the mouse uterus and extensively expressed in endometrial carcinoma.” Tumour Biol 32(2): 359-366).
  • SRSFio has been shown to associate with the translocation liposarcoma protein (TLS) and Ewing's sarcoma protein (EWS) in normal cells.
  • TLS translocation liposarcoma protein
  • EWS Ewing's sarcoma protein
  • the association between SRSFio and TLS has been shown to influence the splicing of adenovirus EiA pre-mRNA (Yang, L., L. J. Embree, et al. (1998). "Oncoprotein TLS interacts with serine-arginine proteins involved in RNA splicing.” J Biol Chem 273(43): 27761-27764).
  • the loss of SRSFio splicing activity has also been suggested in certain cancers.
  • SRSFio has also been linked to the splicing efficiency of the low-density lipoprotein receptor (LDLR) which is involved in cholesterol homeostasis (Ling, I. F. and S. Estus (2010). "Role of SFRS13A in low-density lipoprotein receptor splicing.” Hum Mutat 31(6): 702-709) and has been identified as a master regulatory involved in gene regulatory networks in cell culture models of advanced atherosclerosis (Bjorkegren, J. L., S. Hagg, et al. (2014). "Plasma cholesterol-induced lesion networks activated before regression of early, mature, and advanced atherosclerosis.” PLoS Genet 10(2): 61004201).
  • LDLR low-density lipoprotein receptor
  • SRSFio is encoded by the human gene identified as EntrezGene # 10772 and is located at chromosomal location ip36.11. SRSFio is also known as neural-salient SR protein (NSSR); TLS-associated protein with SR repeats (TASR); splicing factor SRp38; TLS- associated serine-arginine protein 1 (TASRi); TLS-associated serine-arginine protein 2 (TASR2); FUS interacting protein (serine-arginine rich) 1 (FUSIPl); FUS interacting protein (serine-arginine rich) 2 (FUSIP2); splicing factor, arginine/serine-rich 13 (SFRS13); serine- arginine repressor protein 4okDa (SRrp4o); splicing factor arginine/serine-rich 13A
  • NSSR neural-salient SR protein
  • TASRi TLS-associated protein with SR repeats
  • SFRS13A protein phosphatase 1, regulatory subunit 149
  • PPP1R149 protein phosphatase 1, regulatory subunit 149
  • the human transformer 2 beta homolog (hTRA2p) is also a member of the serine/arginine protein family that plays a role in mRNA processing and can be expressed as 5 separate mRNA isoforms through alternative splicing.
  • I1TRA2 ⁇ controls the alternative splicing of the calcitonin-calcitonin gene-related peptide(Tran, Q., T. P. Coleman, et al. (2003).
  • Human transformer 2beta and SRp55 interact with a calcitonin- specific splice enhancer.” Biochim Biophys Acta 1625(2): 141-152), SMN2 (Hofmann, Y., C. L. Lorson, et al. (2000).
  • hTRA2 ⁇ is upregulated in response to hypoxic/oxidative stress and in a variety of disease conditions including cancer (Tsukamoto, Y., N. Matsuo, et al. (2001). "Expression of a novel RNA- splicing factor, RA30i/Tra2beta, in vascular lesions and its role in smooth muscle cell proliferation.”
  • Am J Pathol 158(5): 1685-1694 arteriosclerosis (Tsukamoto, Y., N. Matsuo, et al. (2001).
  • Am J Pathol 158(5): 1685-1694 age-related macular degeneration
  • I1TRA2 ⁇ A role of I1TRA2 ⁇ has also been shown in HIV replication.
  • digoxin was found to suppress HIV gene expression through alteration in the use of specific ⁇ pre- mRNA splice sites. Protein analysis found that digoxin treatment leads to increased abundance of a higher MW form of hTRA2 ⁇ . Overexpression of hTRA2 ⁇ resulted in a reduction in all HIV RNA transcripts (Wong, R. W., A. Balachandran, et al. (2013). "Digoxin suppresses HIV-i replication by altering viral RNA processing.” PLoS Pathog 9(3):
  • SMNi survival motor neuron gene 1
  • the human genome contains a nearly identical copy of the SMNi gene, known as SMN2, yet this gene contains a single nucleotide difference in exon 7, and produces a truncated, less stable SMN protein that is unable to functionally correct for the loss of SMNi (Lorson, C. L., E. Hahnen, et al. (1999). "A single nucleotide in the SMN gene regulates splicing and is responsible for spinal muscular atrophy.” Proc Natl Acad Sci U S A 96(11): 6307-6311).
  • SRp30c-dependent stimulation of survival motor neuron (SMN) exon 7 inclusion is facilitated by a direct interaction with hTra2 beta 1.”
  • Hum Mol Genet 11(5): 577-58 or inhibit the inclusion of exon7 (Wee, C. D., M. A Havens, et al. (2014). "Targeting SR proteins improves SMN expression in spinal muscular atrophy cells.”
  • PLoS One 9(12): 6115205 As described by Hofmann and colleague, increased expression of I1TRA2 ⁇ promotes the inclusion of exon 7 into the SMN2 transcript, leading to increased production of functional SMN protein.
  • Compounds described herein may be useful in the treatment of spinal muscular atrophy. Alternatively, compounds described herein may be useful in the treatment of other neurodegenerative diseases.
  • the human transformer 2 beta homolog (hTRA2 ⁇ ) is encoded by the human gene identified as EntrezGene # 6434 and is located at chromosomal location 3q26.2-q27.
  • hTRA2 ⁇ is also known as SFRS10; SRFS10; TRAN2B; PPP1R156; TRA2-BETA; hTRA2-betai and Htra2- beta.
  • Compounds as described herein may be in the free form or in the form of a salt thereof.
  • compounds as described herein may be in the form of a pharmaceutically acceptable salt, which are known in the art (Berge S. M. et al., J. Pharm. Sci. (1977) 66(i):i- 19).
  • Pharmaceutically acceptable salt as used herein includes, for example, salts that have the desired pharmacological activity of the parent compound (salts which retain the biological effectiveness and/or properties of the parent compound and which are not biologically and/or otherwise undesirable).
  • Compounds as described herein having one or more functional groups capable of forming a salt may be, for example, formed as a
  • compositions containing one or more basic functional groups may be capable of forming a pharmaceutically acceptable salt with, for example, a pharmaceutically acceptable organic or inorganic acid.
  • Pharmaceutically acceptable salts may be derived from, for example, and without limitation, acetic acid, adipic acid, alginic acid, aspartic acid, ascorbic acid, benzoic acid, benzenesulfonic acid, butyric acid, cinnamic acid, citric acid, camphoric acid, camphorsulfonic acid, cyclopentanepropionic acid, diethylacetic acid, digluconic acid, dodecylsulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, glucoheptanoic acid, gluconic acid, glycerophosphoric acid, glycolic acid, hemisulfonic acid, heptanoic acid, hexanoic acid, hydrochloric acid, hydrobromic acid, hydri
  • Compounds containing one or more acidic functional groups may be capable of forming pharmaceutically acceptable salts with a pharmaceutically acceptable base, for example, and without limitation, inorganic bases based on alkaline metals or alkaline earth metals or organic bases such as primary amine compounds, secondary amine compounds, tertiary amine compounds, quaternary amine compounds, substituted amines, naturally occurring substituted amines, cyclic amines, quanidino compounds, or basic ion-exchange resins.
  • inorganic bases based on alkaline metals or alkaline earth metals or organic bases such as primary amine compounds, secondary amine compounds, tertiary amine compounds, quaternary amine compounds, substituted amines, naturally occurring substituted amines, cyclic amines, quanidino compounds, or basic ion-exchange resins.
  • Pharmaceutically acceptable salts may be derived from, for example, and without limitation, a hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation such as ammonium, sodium, potassium, lithium, calcium, magnesium, iron, zinc, copper, manganese or aluminum, ammonia, benzathine, meglumine, methylamine, dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine, isopropylamine, tripropylamine, tributylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, glucamine, methylglucamine, theobromine, purines, piperazine, piperidine, procaine, N- ethylpiperidine, the
  • compounds as described herein may contain both acidic and basic groups and may be in the form of inner salts or zwitterions, for example, and without limitation, betaines.
  • Salts as described herein may be prepared by conventional processes known to a person skilled in the art, for example, and without limitation, by reacting the free form with an organic acid or inorganic acid or base, or by anion exchange or cation exchange from other salts. Those skilled in the art will appreciate that preparation of salts may occur in situ during isolation and purification of the compounds or preparation of salts may occur by separately reacting an isolated and purified compound.
  • compounds and all different forms thereof e.g.
  • Solvates contain either stoichiometric or non-stoichiometric amounts of a solvent in physical association the compound or salt thereof.
  • the solvent may be, for example, and without limitation, a pharmaceutically acceptable solvent.
  • hydrates are formed when the solvent is water or alcoholates are formed when the solvent is an alcohol.
  • compounds and all different forms thereof may include crystalline and amorphous forms, for example, polymorphs, pseudopolymorphs, conformational polymorphs, amorphous forms, or a combination thereof.
  • Polymorphs include different crystal packing arrangements of the same elemental composition of a compound. Polymorphs usually have different X-ray diffraction patterns, infrared spectra, melting points, density, hardness, crystal shape, optical and electrical properties, stability and/or solubility. Those skilled in the art will appreciate that various factors including recrystallization solvent, rate of crystallization and storage temperature may cause a single crystal form to dominate.
  • compounds and all different forms thereof include isomers such as geometrical isomers, optical isomers based on asymmetric carbon, stereoisomers, tautomers, individual enantiomers, individual diastereomers, racemates, diastereomeric mixtures and
  • compositions as described herein may comprise a salt of such a compound, preferably a pharmaceutically or physiologically acceptable salt.
  • compositions will typically comprise one or more carriers, excipients or diluents acceptable for the mode of administration of the preparation, be it by injection, inhalation, topical administration, lavage, or other modes suitable for the selected treatment.
  • Suitable carriers, excipients or diluents are those known in the art for use in such modes of administration.
  • Suitable pharmaceutical compositions may be formulated by means known in the art and their mode of administration and dose determined by the skilled practitioner.
  • parenteral administration a compound may be dissolved in sterile water or saline or a pharmaceutically acceptable vehicle used for administration of non-water soluble compounds such as those used for vitamin K.
  • enteral administration the compound may be administered in a tablet, capsule or dissolved in liquid form.
  • the tablet or capsule may be enteric coated, or in a formulation for sustained release.
  • Many suitable formulations are known, including, polymeric or protein microparticles encapsulating a compound to be released, ointments, pastes, gels, hydrogels, or solutions which can be used topically or locally to administer a compound.
  • a sustained release patch or implant may be employed to provide release over a prolonged period of time.
  • Many techniques known to one of skill in the art are described in Remington: the Science & Practice of Pharmacy by Alfonso Gennaro, 20 th ed., Lippencott Williams & Wilkins, (2000).
  • Formulations for parenteral administration may, for example, contain excipients, polyalkylene glycols such as polyethylene glycol, oils of vegetable origin, or hydrogenated naphthalenes. Biocompatible, biodegradable lactide polymer,
  • lactide/glycolide copolymer or polyoxyethylene-polyoxypropylene copolymers maybe used to control the release of the compounds.
  • Other potentially useful parenteral delivery systems for modulatory compounds include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes.
  • Formulations for inhalation may contain excipients, for example, lactose, or may be aqueous solutions containing, for example, polyoxyethylene-9-lauryl ether, glycocholate and deoxycholate, or may be oily solutions for administration in the form of nasal drops, or as a gel.
  • Compounds or pharmaceutical compositions as described herein or for use as described herein may be administered by means of a medical device or appliance such as an implant, graft, prosthesis, stent, etc.
  • a medical device or appliance such as an implant, graft, prosthesis, stent, etc.
  • implants may be devised which are intended to contain and release such compounds or compositions.
  • An example would be an implant made of a polymeric material adapted to release the compound over a period of time.
  • an “effective amount” of a pharmaceutical composition as described herein includes a therapeutically effective amount or a prophylactically effective amount.
  • a “therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result, such as reduced tumor size, increased life span or increased life expectancy.
  • a therapeutically effective amount of a compound may vary according to factors such as the disease state, age, sex, and weight of the subject, and the ability of the compound to elicit a desired response in the subject. Dosage regimens may be adjusted to provide the optimum therapeutic response.
  • a therapeutically effective amount is also one in which any toxic or detrimental effects of the compound are outweighed by the therapeutically beneficial effects.
  • prophylactically effective amount refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result, such as smaller tumors, increased life span, increased life expectancy or prevention of the progression of prostate cancer to an androgen-independent form.
  • a prophylactically effective amount refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result, such as smaller tumors, increased life span, increased life expectancy or prevention of the progression of prostate cancer to an androgen-independent form.
  • a prophylactic result such as smaller tumors, increased life span, increased life expectancy or prevention of the progression of prostate cancer to an androgen-independent form.
  • prophylactic dose is used in subjects prior to or at an earlier stage of disease, so that a prophylactically effective amount maybe less than a therapeutically effective amount.
  • dosage values may vary with the severity of the condition to be alleviated.
  • specific dosage regimens may be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions.
  • Dosage ranges set forth herein are exemplary only and do not limit the dosage ranges that may be selected by medical practitioners.
  • the amount of active compound(s) in the composition may vary according to factors such as the disease state, age, sex, and weight of the subject. Dosage regimens may be adjusted to provide the optimum therapeutic response. For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It may be advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage.
  • compounds and all different forms thereof as described herein may be used, for example, and without limitation, in combination with other treatment methods for inhibiting HIV infection.
  • Toxicity of the compounds as described herein can be determined using standard techniques, for example, by testing in cell cultures or experimental animals and determining the therapeutic index, i.e., the ratio between the LD50 (the dose lethal to 50% of the population) and the EC50 (The concentration of an investigational drug required to inhibit virus replication by 50 percent). In some circumstances however, such as in severe disease conditions, it may be appropriate to administer substantial excesses of the compositions. Some compounds as described herein may be toxic at some concentrations. Titration studies may be used to determine toxic and non-toxic concentrations. Toxicity may be evaluated by examining a particular compound's or composition's specificity across cell lines. Animal studies may be used to provide an indication if the compound has any effects on other tissues.
  • Compounds as described herein may be administered to a subject.
  • a subject As used herein, a
  • subject may be a human, non-human primate, rat, mouse, cow, horse, pig, sheep, goat, dog, cat, etc.
  • the subject may be suspected of having or at risk for having an HIV infection.
  • NelfinavirTM, EfavirenzTM and 3'-Azido-3'-deoxythymidine (ACTTM) were synthesized by Sigma-AldrichTM (St. Louis, MO). MaravirocTM, RaltegravirTM, ElevategravirTM and
  • DolutegraverTM were obtained from Cedarlane LaboratoriesTM (Ontario, Canada). Indicator cell lines CEM-GXR and GXR-5 were obtained from Dr. Mark Brockman (Simon Fraser University). (N)NRTI-resistant virus (E00443) was obtained from Dr. Zabrina Brumme (Simon Fraser University). HIV-i CXCR4-tropic laboratory-adapted strain NL4-3, and IIIB, HIV-i CCR5-tropic BaL, integrase inhibitor resistant virus (cat. #. 11845), protease inhibitor resistant virus (cat. #. 2948), and HIV-i subtype A (cat. #. 4114) were obtained through the NIH AIDS Research and Reference reagent program, Division of AIDS, NIAID, NIH).
  • HeLa cells were co-transfected with a plasmid carrying Bcl-x minigene X2.13 and a plasmid containing the CMV promoter driving the expression of the SRSFi or SRSF9 cDNA, as described previously in Cloutier, P., J. Toutant, et al. (2008). "Antagonistic effects of the SRp30c protein and cryptic 5' splice sites on the alternative splicing of the apoptotic regulator Bcl-x." J Biol Chem 283(31): 21315-21324, except the present assay was performed in the presence of various concentrations of C8.
  • the anti-HIV activity of C8 was measured by a human T-cell reporter assay based on CEM- GXR cells. These cells express GFP due to the activation of the tat-dependent promoter upon HIV-i infection, and the level of infection was monitored using Flow cytometric analysis (guavaSoft 2.2TM software, Guava HT8, MilliporeTM). Antiviral activity was evaluated in the assay that measure inhibition of HIV-i spread in a co-culture of CEM-GXR cells containing one percent of HIV-i infected (GFP positive) cells. Infection was performed in a 96-well plate containing serial dilutions of molecule C8, and the inhibitory concentration 50 percent (IC50) were determined on day 3.
  • the ViaCount assay was performed with the Guava easyCyte 8HTTM flow cytometer according to the manufacturer's manual (Guava TechnologiesTM, Inc., Hayward, CA, USA). In brief, CEM-GXR cells were seeded in 96-well plates 80000 cells/well in the absence or presence of compound C8 in various concentrations. Twenty-four hours later, 25 ⁇ . of cell suspension was mixed with 225 ⁇ , of Guava ViaCountTM reagent, and the mixture was incubated for 5 min at room temperature. Sample acquisition and data analysis were performed with the selection of EasyFit analysis feature using the ViaCountTM software module.
  • quantitative RT-PCR A total of 200 ng of RNA measured for integrity (using the Agilent Lab-on-ChipTM station) and quantification (using the Thermo Scientific NanoDropTM) was reverse transcribed using random hexamers with Transcriptor Reverse transcriptase in a final volume of 10 ul.
  • HIV-SS.6.REV 5 TGAAGAGGCACAGGCTCCG -3'
  • Reactions were carried out in the ABI 7500 qPCR (Applied BiosystemsTM, Foster City, CA, USA). A first cycle of 10 minutes at 95°C was followed by 40 cycles of 15 seconds at 94°C, 20 seconds at 55°C and 20 seconds at 68°C. Fluorescence measurement using SYBR Green was performed and values were normalized to a set of housekeeping genes.
  • cDNA Twenty ng of cDNA were amplified with 0.2 U/ 10 ul of HotStarTaqTM DNA Polymerase (QiagenTM) in the buffer provided by the manufacturer, and in the presence of the specific primers (IDT) for each splicing unit (at concentrations ranging from 0.3 to 0.6 uM) and dNTPs.
  • a first cycle of 15 minutes at 95°C was followed by 35 cycles of 30 seconds at 94°C, 30 seconds at 55°C and 1 minute at 72°C. Thermocycling was concluded with an extension step of 10 minutes at 72°C. Visualization and analysis of amplified products were done using the LabChip HTTM DNA assay on a Caliper LC-90 automated microfluidic station (CaliperTM, Hopkmton, MA, USA).
  • Plasmids expressing Bcl-x reporter mini genes (X2.13 and X2) and SRSF9 (aka SRp30c) and SRSFi (aka AFS/SF2) were previously described (Garneau D. et al. J Biol Chem. 2005 28o(24):2264i-5o; Shkreta L. et al. J Biol Chem. 2011 286( ⁇ ):33 ⁇ -4 ⁇ , 2011; Cloutier, P., J. Toutant, et al. J Biol Chem. 2008 283(31): 21315-21324).
  • Plasmid transfections were carried out with
  • siRNA used to knock down the expression of SRSFio siGENOME SMARTpool-Human SRSFio, was purchased from DharmaconTM (GE HealthcareTM) and transfected into cells at a concentration of 100 nM using Lipofectamine 2000 (InvitrogenTM). Proteins or RNA were extracted from mock-transfected and siRNA-transfected cells at 72 hours post-transfection.
  • HeLa-HIV cells expressing Flag-SRSFio and treated or not with various concentration of C8 were cultured in 150 mm plates. Collected cells were washed twice with ice-cold PBS and lysed on ice for 30 minutes in NET-2 buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.05 % (vol/vol) Nonidet P-40 added with EDTA-free protease and phosphatase inhibitors cocktail (Roche Diagnostics GmbHTM). The lysates were clarified by centrifugation at 13,000 ⁇ g for 15 min and RNase A solution (0.1 mg/mL cellular lysate) was added.
  • NET-2 buffer 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.05 % (vol/vol) Nonidet P-40 added with EDTA-free protease and phosphatase inhibitors cocktail (Roche Diagnostics GmbHTM).
  • the lysates were clarified by centri
  • the reaction was stopped by adding formic acid to 1% final concentration.
  • the supernatant was transferred to a new tube, while beads were resuspended in 50 of a solution containing 60% acetonitrile, 0.1% formic acid and incubated for 5 min at room temperature. Both supernatants were pooled and lyophilized using a SpeedvacTM.
  • Peptides were resuspended in 30 ⁇ . of 0.1% of trifluoroacetic acid and proceeded to desalting using ZipTip C18 (MilliporeTM) as recommended by the manufacturer. Eluted peptides were lyophilized and resuspended in 25 iL of 1% formic acid.
  • RNA from cells treated with various concentrations of C8 in triplicates (o, 1, 5, 10 and 20 ⁇ ) was purified using the RNAeasyTM mini kit (QiagenTM). A total of 10 ⁇ g of RNA was used to generate an TruSeqTM library using the manufacturer's protocol (iUuminaTM). The library was then sequenced for 51 cycles on a Alumina HiSeq-2500TM platform following the manufacturer's recommendations (AluminaTM).
  • the 51-nucleotide (nt) reads from the sequencer Fastq files were then aligned to the human genome (hgig) with TopHatTM v2.o.io, using the options -p6—no-coverage-search with a transcriptome index produced from the Ensemble GRCI137 release 73 gene build.
  • 51-nt reads were generated for the control, and C8-treated samples. On average, 20M reads were obtained per sample with greater than 95% of reads successfully aligned and 85% of the total reads uniquely aligned to the human genome.
  • RPKM Reads per kilobase of transcript per million mapped reads
  • per genes were then produced by counting the number of uniquely mappable reads that fell within exonic region for all the genes annotated in the Ensembl GRCI137 release 73 build. The total length of exonic regions were used to normalize read counts per gene.
  • the coverage plots were produced by breaking down the spliced transcript in 100 bins and by computing the median read coverage within each bin relative to the total coverage along the transcript.
  • the relative coverage for the different samples was then plotted along the length of the transcript using the same scale for the y axis.
  • the average coverage along all transcripts was calculated by computing the means for each of the 100 bins for all expressed transcripts (transcripts with an RPKM greater than 0.5 in at least two samples) and plotting the resulting coverage for all 100 bins. MISO analysis was performed using the MISO software.
  • TABLE 1 provides a list of the compounds that have been made and an indication of the activity where the compounds have been tested.
  • N-(5-Nitrobenzo[d]isothiazol-3-yl)-4-oxo-i,4-dihydropyridine-3-carboxamide (7) A mixture of acid 2 (8iomg, 5.8mmol) in DCM (6ml) Et 3 N (1.0ml) was cooled to o°C, treated with SOCI 2 (0.52ml, i.2eq) and stirred for 2h. Then the excess of S0C1 2 and DCM were removed under reduced pressure, and the solid was dried for an additional hour.
  • dmydropyridine-3-carboxamide (16) Compound 16 was synthesized by conversion of acid 14 to the corresponding acid fluoride, and reaction of this intermediate with the N- trimethylsilyl derivative of 5-nitrobenzo[d]isothiazol-3-amine.
  • the acid fluoride was diluted with CH 3 CN (3.0 mL) and it was added to the silylated amine at room temperature. After stirring the orange mixture overnight, the solvent was removed under reduced pressure and the crude mixture was flash silica column chromatographed (gradient of 0-50% MeOH in EtOAc) to provide compound 16 as an orange solid (35 mg, 28%).
  • Example 1 Anti-HIV activity of compounds C8, E5, D3, and C2.
  • GXR-5 human T-cell-based reporter cell line (GXR-5) that expresses the HIV-i receptor CD4, the CXCR4 and CCR5 co-receptors was used, and green fluorescent protein (GFP) as a direct and quantitative marker, as described previously (Brockman et al. 2006 J. Virol Methods 131(2): 134-142).
  • GXR-5 cells express GFP due to the activation of the tat-dependent promoter upon HIV infection, and the level of infection can be monitored using Flow cytometric analysis (guavaSoft 2.2TM software, Guava HT8TM, MilliporeTM).
  • a co-culture method using infected and non-infected cells was used to assay both viral replication and cell to cell spread, as described previously (Chen et al. 2007. J. Virol 8i(22):i2582-95). Briefly, a co-culture of lxio 6 non-infected GXR-5 cells and 1% HIV- 1NL4-3 infected cells was exposed to varying concentrations of the test compound in DMSO or 1 uM Nelfinavir as a control. At four days post-infection, the level of infection was determined by flow-cytometry using GFP expression as a read-out.
  • FIGURE lA the C8 and D3 compounds showed a dose-dependent ability to inhibit viral replication and cell to cell spread, whereas compounds E5 and C2 showed the same level of anti-HIV activity regardless of the dose used.
  • SL309 showed a gradual dose-dependent ability to inhibit viral replication.
  • FIGURE 15 shows an inhibition of viral spread at various concentrations of SL309 from o ⁇ to 25 ⁇ .
  • Example 2 The antiretroviral activity of C8 on wild-type HIV-i and drug resistant strains
  • C8 was evaluated for its antiretroviral activity using a human T-cell line (CEM-GXR) against wild-type HIV-i in both subtype A and subtype B, and against multiple drug-resistant strains ofHIV-l.
  • CEM-GXR human T-cell line
  • CEM-GXR cells express HIV-i receptor CD4, both the CXCR4 and CCR5 coreceptors, and GFP as a direct and quantitative marker.
  • CEM-GXR cells express GFP due to the activation of the tat-dependent promoter upon HIV infection, and the level of infection was monitored using Flow cytometric analysis (guavaSoft 2.2TM software, Guava HT8TM, MilliporeTM).
  • Antiviral activity was evaluated in the assay that measures inhibition of HIV-i spread in a co- culture of one million CEM-GXR cells containing one percent HIV-i variants infected cells. Infection was performed in a T25 flask containing 5ml RPMI 1640 supplemented with 10% FBS. C8 is then added to the culture at final concentration from ⁇ . ⁇ -8. ⁇ from a 5mM stock. At four days post-infection, the level of infection is determined by flow cytometry using GFP expression as readout.
  • C8 showed activity at the submicromolar concentrations against both HIV-IIIIB (subtype B, X4-tropic), and HIV-i 97USSN54 (subtype A, Rs-tropic), with IC 50 of ⁇ . ⁇ and 0.9UM respectively (FIGURE 2A and B).
  • NRTI Non-Nucleoside Reverse Transcriptase Inhibitor
  • PI Protease Inhibitor
  • II Integrase Inhibitor
  • This virus also carries the mutations D67N, K70R, V75I, F77L, Y115F, F116Y, Q151M, M184V, T215V, K219Q which confer for high-level resistance to lamivudine (3TC), abacavir (ABC), zidovudine (AZT), stavudine (D4T) , didanosine (DDI) , emtricitabine (FTC), tenofovir (TDF). C8 showed activity against this (N)NRTI-resistant virus with an IC 50 of ⁇ .3 ⁇ (FIGURE 2C).
  • C8 also displayed potency against a Pi-resistant isolate (2948) that contains substitution G48V, and L90M. This combination of mutation decreases susceptibility to each of the FDA approved Pis except Tipranavir (TPV) and Darunavir (DRV) (Rhee et al 2010 Antimicrob Agents Chemother). C8 retained its activity against this Pi-resistant strain with an IC 50 of ⁇ .4 ⁇ (FIGURE 2D).
  • HIV-i chnical isolate resistant to integrase inhibitors (11845) was tested for its susceptibility to C8.
  • This virus harbors G140S, and Q148H substitution that reduce Raltegravir (RAL) and Elvitegravir (EVG) susceptibility > 100-fold (Abram et al. 2013 Antimicrob Agents
  • FIGURE 3 shows four active molecules (i.e. D3, E5, C2, and C8) in 2 ⁇ final concentration for their ability to inhibit HIV-i replication of HAART RTI-resistant strains.
  • the resistant strains used were the highly resistant Eoo443_v, the moderately resistant Eoo435_4 and the slightly resistant E oo476_v.
  • DGPS56/DH-052 C8
  • DGPS39/LB-45 E5
  • DGPS154/MM-99 D3
  • DGPS73/SKK-31 C2
  • Nelfinavir ⁇ a protease inhibitor more potent than AZT
  • FIGURE 3 summarizes the results of this experiment.
  • the ability of DGPS56/DH-052 and DGPS154/ M-99 to inhibit the virus increased as their concentration increased.
  • the change in concentration had no significant effect on the inhibition of HIV-i replication. This could be due to the fact that the difference between concentrations was not large enough to observe a significant difference.
  • DGPS56/DH-052 appeared to be the most potent.
  • Example 3 Compound C8 interferes with the splicing of HIV transcripts
  • HeLa rtTA-HV- AMls A HeLa cell line stably expressing a modified X4HIV-1 (LAQ+I) provirus (HeLa rtTA-HV- AMls) was used to determine the effect of compound C8 on the splicing of HIV transcripts and the transcription of the HIV genome as described previously by Wong et al. (Wong et al.
  • Protein levels for the Gag protein were determined by ELISA according to the manufacturer's protocol. Protein levels for the Env and Tat proteins were determined by Western blot analysis and fluorescent microscopy. As shown in FIGURE 4B and 4C, quantitative RT-PCR determined that C8 at various concentrations (i.e. 1 ⁇ , 5 ⁇ and 10 ⁇ ) induced a marked decrease in the level of HIV-i RNA accumulation in HeLa-H cells (of selected amplicons as shown in FIGURE 4A). The intensity of products was normalized relative to the amount produced in the controls (which corresponds to an arbitrary unit value of 1).
  • FIGURE 5C shows the structure of the HIV-i provirus used to generate the HeLa cell line with the GagGFP fusion, wherein the provirus has elements (TetO sites, rtTA) required for regulation by doxycycline (Dox). Similarly, the negative impact of C8 on the production the Gag viral protein was further confirmed by fluorescent microscopy fluroscent microscopy.
  • FIGURE 5D shows the effect of C8 on HIV-i US RNA localization
  • the cells were incubated in the absence (-) or the presence (+) of Dox for 24 h, then fixed, stained with DAPI and imaged using a Leica DMR epifluorescent microscope and HeLa HIVGagGFP cells were treated with either DMSO or C8 (1 ⁇ ) for 4 h prior to addition of Dox to induce provirus expression, wherein 24 hours after Dox addition, cells were fixed and US RNA localization was determined by in situ hybridization
  • HIV-i splicing was analyzed by quantitative RT-PCR using different combinations of forward and reverse primers to amplify products corresponding to various splicing events (see the map of donor and acceptor splice sites used for the production of the major HIV mRNAs as shown in FIGURE 6A), wherein the AS2 set of primers was used to monitor the production of Tati and Nef2 in panels FIGURES B and C.
  • the differential impact of C8 on products derived from different splicing events indicates that these effects are not likely due to a reduction in HIV transcription alone, and is therefore consistent with the notion that C8 differentially affects splice site usage in HIV-i.
  • FIGURE 6B shows the results from the endpoint RT-PCR assay from HeLa-HIV cells treated with C8.
  • FIGURE 6C shows the histograms depicting the impact of C8 on Tati and Nef2 production, wherein the intensity of products was normalized relative to the total intensity for each lane.
  • FIGURES 7A shows that compound C8 at 1 uM, 5 uM, 10 uM did not significantly affect the level of unspliced RNA in the HeLa-HIV cell line with three different primer pairs (US-i, US-2 and US-4).
  • FIGURE 7B shows a dose dependent reduction of Tat transactivated HIV promoter activity in response to compound C8 at 1 uM, 10 ⁇ , 20 uM, measured as total GFP fluorescence at 24 hours post-transfection.
  • Example 4 SRSF10 is a host target of the C8 compound
  • Bcl-x is alternatively spliced through the use of two 5' splice sites to produce the Bcl-xL and Bcl-xS splice variants (Cloutier, P., J. Toutant, et al. (2008). J Biol Chem 283(31): 21315-21324).
  • a control we used a plasmid that expresses SRSF9 (aka SRp30c), a protein that also regulates Bcl-x splicing (Cloutier, P., J. Toutant, et al. (2008). J Biol Chem 283(31): 21315-21324).
  • a Flag-tagged SRSFio protein stimulates the production of pro- apoptotic Bcl-xS on transcripts derived from a Bcl-x mini-gene (X2), which contains different control elements than X2.13 (FIGURE 8B).
  • X2 Bcl-x mini-gene
  • 10 ⁇ of C8 completely abrogated the SRSFio-induced shift in splicing (FIGURE 8B). Since C8 had little effect on the expression of the Flag-SRSFio protein (FIGURE 8C), our results indicate that C8 had selectively abrogated the splicing regulatory function of SRSFio.
  • Example 5 SRSFio controls HIV splicing and C8 affects the phosphorylation of SRSFio
  • HIV-i splicing should be sensitive to a reduction in the level of SRSFio.
  • an siRNA to partially deplete SRSFio in the HeLa-HIV cell line was used (FIGURE 9A). Quantitative RT-PCR analysis indicated that this partial depletion of SRSFio reduced by 20 to 30 percentage points the level of products derived from the D4/A7 splice (SS5 and SS6 pairs of primers), the D1/A5 splice (SSi and SS1.2 pairs of primers), the D1/A2 splice (SS3 pair of primers) and the D1/A4 splice (SS4 pair of primers).
  • siRNA against SRSFio had no impact on HIV transcription as measured by products derived from the unspliced genomic pre-mRNA (USi, US2 and US4 pairs of primers) (FIGURE 9B). These results indicate that SRSFio participates in the control of HIV-i splicing. The impact of C8 on HIV-i splicing may therefore be caused, at least in part, by an alteration in the activity of SRSFio.
  • SRSFio is a phosphorylated protein, and its dephosphorylation converts SRSFio from a positive regulator into a repressor (Shin, C, Y. Feng, et al. (2004). "Dephosphorylated SRp38 acts as a splicing repressor in response to heat shock.” Nature 427(6974): 553-558; and Feng, Y., M. Chen, et al. (2008). "Phosphorylation switches the general splicing repressor SRp38 to a sequence-specific activator.” Nat Struct Mol Biol 15(10): 1040-1048).
  • Treating extracts with calf intestinal phosphatase produces a dephosphorylated version of SRSFio with distinctively faster gel mobility than phosphorylated forms(Shin, C. and J. L. Manley (2002). "The SR protein SRp38 represses splicing in M phase cells.” Cell 111(3): 407-417). This associated change in migration was reproduced when we treated extracts from cells expressing FlagSRSFio and HA-tagged SRSFio proteins with CIP and fractionated proteins using appropriate gel-migrating conditions (FIGURE 10A).
  • phosphorylation event might be altered by C8.
  • An anti-HIV compound that alters the activity of a cellular protein may be of limited use because it may affect the normal cellular functions carried out by the cellular protein.
  • C8 had a modest negative impact on cell growth (FIGURE 12), it was of interest to determine whether C8 also affected the alternative splicing of cellular genes.
  • a preliminary screen using RT-PCR was carried out on a set of 96 alternative splicing events from 92 different human genes (data not shown). At 10 ⁇ of C8, only one splicing event (INF2) shifted with a q-value inferior to 0.01. At 1 uM of C8, a concentration that severely blocks HIV replication, no alternative splicing event from this set was significantly altered (q-value ⁇ 0.05).
  • BCLAFi as a gene whose alternative splicing is regulated by SRSFio (Zhou, X., X. Li, et al. (2014). "BCLAFi and its splicing regulator SRSFio regulate the tumorigenic potential of colon cancer cells.” Nat Commun 5: 4581).
  • SRSFio splicing regulator SRSFio regulate the tumorigenic potential of colon cancer cells. Nat Commun 5: 4581.
  • the ability of C8 to impact the alternative splicing of BCLAFi was tested in the HeLa-HIV cell line as well as in three colon cancer cell lines (Caco2, SW620 and HCT116).
  • Example 8 C8 suppresses adenovirus replication
  • A549 cells human carcinomic alveolar basal epithelial cell line
  • the media was removed and replaced with fresh media containing either C8 or DMSO as a control.
  • Cells were harvested after 24 hrs and analyzed by Western blot.
  • FIGURE 14A the presence of ⁇ of C8 resulted in a significant reduction in the levels of the adenovirus hexon protein, suggesting that C8 effectively suppressed the viral replication.
  • Example 9 Treatment with C8 leads to increased production of full length SMN2 transcript
  • hTra2P can promote the inclusion of the survival motor neuron 2 (SMN2) exon 7, leading to full length SMN2 mRNA ( Hofmann, Y., C. L. Lorson, et al. (2000). "Htra2-beta 1 stimulates an exonic splicing enhancer and can restore full-length SMN expression to survival motor neuron 2 (SMN2)." Proc Natl Acad Sci U S A 97(17): 9618-9623).
  • SSN2 survival motor neuron 2
  • endpoint RT-PCR was used to determine whether treatment with C8 affected the inclusion of exon 7 of SMN2 using a cell line derived from a patient with spinal muscular atrophy (SMA).
  • SMA spinal muscular atrophy
  • FIGURE 7C shows no significant change in the transcription on housekeeping cellular genes MRPL19, PUMi and YWHAZ in HeLa-HIV a cell line in response to various concentrations of compound C8 (o, 1, 10 and 20 ⁇ ).
  • a transcriptome analysis was carried out by performing RNA-Seq on total RNA from the HeLa- HIV cells treated with DMSO and increasing concentrations of C8 (o, 1, 5, 10 and 20 ⁇ ) in triplicates.
  • MISO software Karlz, Y., E. T. Wang, et al. (2010). Nat Methods 7(12): 1009- 1015) was used to follow thousands of alternative splicing events.
  • the MISO analysis revealed 66 alternative splicing units affected by more than 30 percentage points with 20 ⁇ of C8 with a Bayesian coefficient score greater than 400 (data not shown). 22 of these units were selected for validation by endpoint RT-PCR using o, 1, 5 and 10 ⁇ of C8 on HeLa-HIV cells. Of the 22 units, only 6 were confirmed as hits by C8 (DOM3Z(DXO), ESYT2, ITGB3BP, NAP1L4, RBM41, SPTANi ; data not shown). Although 10 uM of C8 shifted these six events between 13 and 45 percentage points, all the shifts observed with 1 ⁇ of C8 were between o and 12 percentage points, values that are essentially below or at the validation threshold.

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Virology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • AIDS & HIV (AREA)
  • General Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention porte sur un composé ayant une structure des formules indiquées, et des utilisations de ces composés pour inhiber l'infection par le VIH, l'infection par l'adénovirus, ou pour traiter le cancer du colon ou l'atrophie musculaire spinale.
PCT/CA2015/000291 2014-04-29 2015-04-29 Composés dérivés de benzisothiazole utilisés comme agents thérapeutiques et leurs méthodes d'utilisation Ceased WO2015164956A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201461985963P 2014-04-29 2014-04-29
US61/985,963 2014-04-29

Publications (1)

Publication Number Publication Date
WO2015164956A1 true WO2015164956A1 (fr) 2015-11-05

Family

ID=54357942

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CA2015/000291 Ceased WO2015164956A1 (fr) 2014-04-29 2015-04-29 Composés dérivés de benzisothiazole utilisés comme agents thérapeutiques et leurs méthodes d'utilisation

Country Status (1)

Country Link
WO (1) WO2015164956A1 (fr)

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10266530B2 (en) 2016-09-09 2019-04-23 Incyte Corporation Pyrazolopyridine compounds and uses thereof
US10280164B2 (en) 2016-09-09 2019-05-07 Incyte Corporation Pyrazolopyridone compounds and uses thereof
WO2019192743A1 (fr) 2018-04-04 2019-10-10 INSERM (Institut National de la Santé et de la Recherche Médicale) Méthodes de traitement d'infection à virus de l'hépatite b (hbv)
US10722495B2 (en) 2017-09-08 2020-07-28 Incyte Corporation Cyanoindazole compounds and uses thereof
US10745388B2 (en) 2018-02-20 2020-08-18 Incyte Corporation Indazole compounds and uses thereof
US10752635B2 (en) 2018-02-20 2020-08-25 Incyte Corporation Indazole compounds and uses thereof
US10800761B2 (en) 2018-02-20 2020-10-13 Incyte Corporation Carboxamide compounds and uses thereof
US10899755B2 (en) 2018-08-08 2021-01-26 Incyte Corporation Benzothiazole compounds and uses thereof
US11014929B2 (en) 2016-09-09 2021-05-25 Incyte Corporation Pyrazolopyrimidine compounds and uses thereof
US11066394B2 (en) 2019-08-06 2021-07-20 Incyte Corporation Solid forms of an HPK1 inhibitor
US11111247B2 (en) 2018-09-25 2021-09-07 Incyte Corporation Pyrazolopyrimidine compounds and uses thereof
US11242343B2 (en) 2016-09-09 2022-02-08 Incyte Corporation Pyrazolopyridine compounds and uses thereof
US11299473B2 (en) 2018-04-13 2022-04-12 Incyte Corporation Benzimidazole and indole compounds and uses thereof
US11406624B2 (en) 2017-02-15 2022-08-09 Incyte Corporation Pyrazolopyridine compounds and uses thereof
WO2022178283A1 (fr) * 2021-02-19 2022-08-25 The Regents On The University Of California Compositions et méthodes de traitement de tauopathies

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009079767A1 (fr) * 2007-12-21 2009-07-02 University Health Network Dérivés d'indolmone à substitution indazolyle, benzimidazolyle, benzotriazolyle en tant qu'inhibiteurs de kinases utilisés dans le traitement du cancer

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009079767A1 (fr) * 2007-12-21 2009-07-02 University Health Network Dérivés d'indolmone à substitution indazolyle, benzimidazolyle, benzotriazolyle en tant qu'inhibiteurs de kinases utilisés dans le traitement du cancer

Cited By (30)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11242343B2 (en) 2016-09-09 2022-02-08 Incyte Corporation Pyrazolopyridine compounds and uses thereof
US10280164B2 (en) 2016-09-09 2019-05-07 Incyte Corporation Pyrazolopyridone compounds and uses thereof
US10435405B2 (en) 2016-09-09 2019-10-08 Incyte Corporation Pyrazolopyridine compounds and uses thereof
US12384778B2 (en) 2016-09-09 2025-08-12 Incyte Corporation Pyrazolopyridine compounds and uses thereof
US11891388B2 (en) 2016-09-09 2024-02-06 Incyte Corporation Pyrazolopyridine compounds and uses thereof
US11795166B2 (en) 2016-09-09 2023-10-24 Incyte Corporation Pyrazolopyridine compounds and uses thereof
US10266530B2 (en) 2016-09-09 2019-04-23 Incyte Corporation Pyrazolopyridine compounds and uses thereof
US11542265B2 (en) 2016-09-09 2023-01-03 Incyte Corporation Pyrazolopyrimidine compounds and uses thereof
US10934288B2 (en) 2016-09-09 2021-03-02 Incyte Corporation Pyrazolopyridine compounds and uses thereof
US11014929B2 (en) 2016-09-09 2021-05-25 Incyte Corporation Pyrazolopyrimidine compounds and uses thereof
US11406624B2 (en) 2017-02-15 2022-08-09 Incyte Corporation Pyrazolopyridine compounds and uses thereof
US10722495B2 (en) 2017-09-08 2020-07-28 Incyte Corporation Cyanoindazole compounds and uses thereof
US11731958B2 (en) 2018-02-20 2023-08-22 Incyte Corporation Carboxamide compounds and uses thereof
US10745388B2 (en) 2018-02-20 2020-08-18 Incyte Corporation Indazole compounds and uses thereof
US12466815B2 (en) 2018-02-20 2025-11-11 Incyte Corporation Carboxamide compounds and uses thereof
US10752635B2 (en) 2018-02-20 2020-08-25 Incyte Corporation Indazole compounds and uses thereof
US10800761B2 (en) 2018-02-20 2020-10-13 Incyte Corporation Carboxamide compounds and uses thereof
US11492354B2 (en) 2018-02-20 2022-11-08 Incyte Corporation Indazole compounds and uses thereof
CN112367993A (zh) * 2018-04-04 2021-02-12 Inserm(法国国家健康医学研究院) 治疗乙型肝炎病毒（hbv）感染的方法
JP2021520407A (ja) * 2018-04-04 2021-08-19 インサーム（インスティテュ ナシオナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシェ メディカル）Ｉｎｓｅｒｍ（Ｉｎｓｔｉｔｕｔ Ｎａｔｉｏｎａｌ Ｄｅｌａ Ｓａｎｔｅ Ｅｔ Ｄｅ Ｌａ Ｒｅｃｈｅｒｃｈｅ Ｍｅｄｉｃａｌｅ） Ｂ型肝炎ウィルス（ｈｂｖ）感染を処置する方法
WO2019192743A1 (fr) 2018-04-04 2019-10-10 INSERM (Institut National de la Santé et de la Recherche Médicale) Méthodes de traitement d'infection à virus de l'hépatite b (hbv)
US11299473B2 (en) 2018-04-13 2022-04-12 Incyte Corporation Benzimidazole and indole compounds and uses thereof
US10899755B2 (en) 2018-08-08 2021-01-26 Incyte Corporation Benzothiazole compounds and uses thereof
US11866426B2 (en) 2018-08-08 2024-01-09 Incyte Corporation Benzothiazole compounds and uses thereof
US12441731B2 (en) 2018-09-25 2025-10-14 Incyte Corporation Pyrazolopyrimidine compounds and uses thereof
US11111247B2 (en) 2018-09-25 2021-09-07 Incyte Corporation Pyrazolopyrimidine compounds and uses thereof
US11787784B2 (en) 2019-08-06 2023-10-17 Incyte Corporation Solid forms of an HPK1 inhibitor
US11066394B2 (en) 2019-08-06 2021-07-20 Incyte Corporation Solid forms of an HPK1 inhibitor
WO2022178283A1 (fr) * 2021-02-19 2022-08-25 The Regents On The University Of California Compositions et méthodes de traitement de tauopathies
US20240174625A1 (en) * 2021-02-19 2024-05-30 The Regents Of The University Of California Compositions and methods for treating tauopathies

Similar Documents

Publication Publication Date Title
WO2015164956A1 (fr) Composés dérivés de benzisothiazole utilisés comme agents thérapeutiques et leurs méthodes d'utilisation
ES2934810T3 (es) Moléculas químicas que inhiben el mecanismo de empalme para tratar enfermedades causadas por anomalías de empalme
CN105142642B (zh) 癌症治疗方法
US8816089B2 (en) Methods for controlling SR protein phosphorylation, and antiviral agents whose active ingredients comprise agents that control SR protein activity
JP5247470B2 (ja) ヒストンデアセチラーゼのインヒビターとしてのピリジン及びピリミジン誘導体
JP4725946B2 (ja) ヒストンデアセチラーゼの新規な阻害剤としてのスルホニル誘導体
JP4644428B2 (ja) ヒストンデアセチラーゼの新規な阻害剤
Zhong et al. Identification of benzenesulfonamide quinoline derivatives as potent HIV-1 replication inhibitors targeting Rev protein
US10941121B2 (en) Human helicase DDX3 inhibitors as therapeutic agents
KR20170139055A (ko) 백금계 항암제에 의해 유도된 세포 내 증가된 p53 수준을 유지시키는 방법 및 이의 활용
EA007908B1 (ru) Карбониламинопроизводные как новые ингибиторы гистондеацетилазы
US9757350B2 (en) Small molecule inhibitors of replication protein A that also act synergistically with cisplatin
CN106794125A (zh) 用于治疗或预防核纤层蛋白病、老化和癌症的nat10调节剂
Massari et al. Blocking HIV-1 replication by targeting the Tat-hijacked transcriptional machinery
Wang et al. Design, synthesis, and biological evaluation of 2-Anilino-4-Triazolpyrimidine derivatives as CDK4/HDACs inhibitors
Li et al. Hybrid molecules based on an emodin scaffold. Synthesis and activity against SARS-CoV-2 and Plasmodium
Doan et al. Novel (E)-3-(1-substituted-1 H-indazol-5-yl)-N-hydroxypropenamides as histone deacetylase inhibitors: design, synthesis and structure–activity relationships
US20190308956A1 (en) Piperazine derivatives as antiviral agents with increased therapeutic activity
US10975059B2 (en) Small molecule BRD4 modulators for HIV epigenetic regulation
WO2021127367A2 (fr) Procédés d'inhibition de l'arnm de l'alpha-synucléine à l'aide de petites molécules
JP2023506860A (ja) 抗ウイルス性化合物、組成物及び使用方法
JP2006348021A (ja) 細胞周期阻害剤
Bandy Development of inhibitors of alternative splicing events crucial to HIV-1 replication: A new anti-HIV/AIDS strategy targeting the host cell machinery
BR112017017229B1 (pt) Composto, composto para uso médico ou na preparação de um medicamento e composição farmacêutica

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15785629

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 15785629

Country of ref document: EP

Kind code of ref document: A1