U.S. patent application number 14/856329 was filed with the patent office on 2016-07-21 for broad spectrum antiviral and antiparasitic agents.
The applicant listed for this patent is The Regents of the University of California. Invention is credited to Peter J. Bradley, Bettina E. Hajagos, Michael E. Jung, Benhur Lee, Jihye Lee, Frederic Vigant, Michael C. Wolf.
Application Number | 20160207908 14/856329 |
Document ID | / |
Family ID | 44799297 |
Filed Date | 2016-07-21 |
United States Patent
Application |
20160207908 |
Kind Code |
A1 |
Lee; Benhur ; et
al. |
July 21, 2016 |
BROAD SPECTRUM ANTIVIRAL AND ANTIPARASITIC AGENTS
Abstract
The invention provides compounds represented by the formulae:
(I) and (II) where the compounds are useful in treating, for
example, viral and/or parasitic infections. Also provided are
pharmaceutical compositions comprising the compounds and methods of
treatment using the compounds.
Inventors: |
Lee; Benhur; (Los Angeles,
CA) ; Jung; Michael E.; (Los Angeles, CA) ;
Lee; Jihye; (Los Angeles, CA) ; Vigant; Frederic;
(Hollywood, CA) ; Bradley; Peter J.; (Los Angeles,
CA) ; Wolf; Michael C.; (Washington, DC) ;
Hajagos; Bettina E.; (Corona Del Mar, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
The Regents of the University of California |
Oakland |
CA |
US |
|
|
Family ID: |
44799297 |
Appl. No.: |
14/856329 |
Filed: |
September 16, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13640732 |
Oct 11, 2012 |
9139575 |
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PCT/US2011/032336 |
Apr 13, 2011 |
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14856329 |
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61467912 |
Mar 25, 2011 |
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61323823 |
Apr 13, 2010 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 31/22 20180101;
A61P 31/18 20180101; C07D 413/06 20130101; A61P 33/00 20180101;
C07D 405/06 20130101; A61P 31/12 20180101; A61P 31/00 20180101;
A61K 9/0014 20130101; A61P 31/14 20180101; A61K 9/127 20130101 |
International
Class: |
C07D 413/06 20060101
C07D413/06; A61K 9/00 20060101 A61K009/00; C07D 405/06 20060101
C07D405/06; A61K 9/127 20060101 A61K009/127 |
Goverment Interests
STATEMENT OF GOVERNMENT SUPPORT
[0002] This invention was made with Government support of Grant
Nos. AI 070495 and AI 082100, awarded by the National Institutes of
Health. The Government has certain rights in this invention.
Claims
1. A compound according to formula Ia or a pharmaceutically
acceptable salt thereof, ##STR00058## wherein R.sup.1 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, C.sub.3-C.sub.6 aryl,
C.sub.3-C.sub.6 heteroaryl, C.sub.3-C.sub.6 cycloalkyl, or
C.sub.3-C.sub.6 heterocycloalkyl, each optionally substituted with
halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
R.sup.2 represents one or more substituents independently selected
from --OR.sup.3, --SR.sup.3, and --NR.sup.4R.sup.5; R.sup.3,
R.sup.4, and R.sup.5 are independently H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl,
C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3, --CN,
--OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5; and X is O, N, S, NH, or NR.sup.3.
2. A compound according to formula IIa or a pharmaceutically
acceptable salt thereof, ##STR00059## wherein R.sup.1 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, each optionally substituted with halo, --NO.sub.2,
--CF.sub.3, --CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3,
--NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5; halo, --NO.sub.2, --CF.sub.3, --CN,
--OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5; R.sup.2 represents exactly one
--OR.sup.3, --SR.sup.3, or --NR.sup.4R.sup.5; R.sup.3, R.sup.4, and
R.sup.5 are independently H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, or C.sub.1-C.sub.6 alkoxy, halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; X is
O, N, S, NH, or NR.sup.3; and Z is O, N, NH, or NR.sup.3.
3. The compound of claim 1, wherein R.sup.3, R.sup.4, and R.sup.5
are independently C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
or C.sub.2-C.sub.6 alkynyl.
4. The compound of claim 1, wherein R.sup.1 is C.sub.1-C.sub.6
alkyl.
5. A method of treating or preventing a disease or condition caused
by infection with an enveloped virus, the method comprising
administering to a subject a therapeutically effective amount of
the compound of claim 1.
6. A method of treating or preventing a disease or condition caused
by infection with an enveloped virus, the method comprising
administering to a subject a therapeutically effective amount of a
compound according to formula IIa or a pharmaceutically acceptable
salt thereof, ##STR00060## wherein R.sup.1 is H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl, each
optionally substituted with halo, --NO.sub.2, --CF.sub.3, --CN,
--OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5; halo, --NO.sub.2, --CF.sub.3, --CN,
--OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5; R.sup.2 represents one or more
substituents independently selected from H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6
alkoxy, halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, and --SO.sub.2NR.sup.4R.sup.5;
R.sup.3, R.sup.4, and R.sup.5 are independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3, --CN,
--OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5; X is O, N, S, NH, or NR.sup.3; and Z is
O, N, NH, or NR.sup.3.
7. The method of claim 5, wherein the virus is hepatitis C virus
(HCV), human immunodeficiency virus (HIV), herpes simplex virus
type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), Ebola virus,
Influenza virus, Nipah virus, Yellow Fever Virus, Dengue virus,
Rift Valley Fever Virus, West Nile Virus.
8. The method of claim 5, wherein the compound is administered as a
liposomal formulation.
9. A pharmaceutical composition comprising the compound of claim 1
and at least one pharmaceutically acceptable excipient.
10. The pharmaceutical composition of claim 9, wherein the
composition is suitable as a topical formulation.
11. The pharmaceutical composition of claim 10, which is in the
form of a patch, an ointment, a cream, a lotion, a drop, a spray,
or an aerosol.
12. The pharmaceutical composition of claim 10, which is in the
form of a liposomal formulation.
13. A method of treating or preventing a disease or condition
caused by infection with a parasite, the method comprising
administering to a subject a therapeutically effective amount of
the compound of claim 1.
14. A method of treating or preventing a disease or condition
caused by infection with a parasite, the method comprising
administering to a subject a therapeutically effective amount of a
compound according to formula II or a pharmaceutically acceptable
salt thereof, ##STR00061## wherein R.sup.1 is H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.1-C.sub.6 alkoxy, C.sub.3-C.sub.6 aryl, C.sub.3-C.sub.6
heteroaryl, C.sub.3-C.sub.6 cycloalkyl, or C.sub.3-C.sub.6
heterocycloalkyl, each optionally substituted with halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
R.sup.2 represents one or more substituents independently selected
from --OR.sup.3, --SR.sup.3, and --NR.sup.4R.sup.5; R.sup.3,
R.sup.4, and R.sup.5 are independently H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6
alkoxy, halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; X is
O, N, S, NH, or NR.sup.3; and Z is O, N, NH, or NR.sup.3.
Description
RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Applications 61/323,823 filed Apr. 13, 2010 and 61/467,912 filed
Mar. 25, 2011, both of which are hereby incorporated by reference
in their entirety for all purposes.
BACKGROUND
[0003] There are few licensed and efficacious broad-spectrum
antivirals currently available. Examples include ribavirin, which
functions via nebulous effects on both host and virus proteins, and
alpha-interferon, which produces unwanted side effects and remains
impractically expensive for widespread use (Tam et al., Antivir
Chem Chemother, 12:261-72 (2001); Bekisz et al., Growth Factors,
22:243-51 (2004); de Veer et al., J Leukoc Biol, 69:912-20 (2001);
Sen, Annu Rev Microbiol, 55:255-81 (2001); Hong and Cameron, Prog
Drug Res, 59:41-69 (2002)). The prevailing paradigm in antiviral
research emphasizes a "one bug-one drug" strategy; however, the
rapid rise in the number of emerging viral pathogens brings into
stark contrast the limited resources available to develop
therapeutics on a single-pathogen basis (see e.g., Burroughs et
al., "The Emergence of Zoonotic Diseases: Understanding the Impact
on Animal and Human Health," Workshop Summary from Board on Global
Health, Institute of Medicine, National Academy Press, Washington,
D.C. (2002)). The expense and difficulty of developing antiviral
drugs tailored to specific pathogens underscores the need to
develop broad spectrum antiviral drugs against targets that are
common among large classes of viruses.
[0004] Viruses can be categorized as either lipid-enveloped or
non-enveloped (naked). Enveloped viruses replicate within the
host-cell, recruit viral proteins to the host membrane, and then
bud from and utilize the host membrane, essentially, as a vehicle
to transport the viral genome to new cellular targets. Although the
lipid membrane of enveloped viruses derives from the host cell, it
differs from host cellular membranes in several biochemical and
biophysical properties, such as biogenic reparative capacity,
fluidity, lipid composition, and curvature. For example, the
membranes of budding viral particles are highly curved relative to
the membranes of much larger host cells. As a result, the fusion of
enveloped viral particles with new host cells requires that the
high curvature viral membranes undergo elastic stresses and
subsequent negative curvature needed to promote fusion between the
juxtaposed outer lipid monolayers of the viral particles and host
cell membranes (Chernomordik et al., J Cell Biol, 175:201-7 (2006);
McMahon and Gallop, Nature, 438:590-6 (2005); Chemomordik and
Kozlov, Annu Rev Biochem, 72:175-207 (2003)).
[0005] The central role of virus-host cell fusion in the
infectivity of enveloped viruses has motivated the development of
small molecule antiviral therapeutics that insert, intercalate, or
otherwise bind to viral membranes and disturb the membrane dynamics
required for successful virus-host cell fusion (e.g., Chemomordik
et al., J Cell Biol, 175:201-7 (2006); Martin and Ruysschaert,
Biochim Biophys Acta, 1240:95-100 (1995); Langosch et al., J Biol
Chem, 276:32016-21 (2001); Langosch et al., Cell Mol Life Sci,
64:850-64 (2007)). For example, the phospholipid analog
lysophosphotidylcholine (LPC) is designed to prevent the entry of
certain enveloped viruses, such as influenza, HIV-1 (Class I
fusion) and TBEV (Class II fusion), into host cell by stabilizing
the positive spontaneous curvature of viral membranes and thereby
preventing conformational changes needed for viral-host cell fusion
(Chernomordik et al., J Cell Biol, 175:201-7 (2006); Chernomordik
and Kozlov, Annu Rev Biochem, 72:175-207 (2003); Martin and
Ruysschaert, Biochim Biophys Acta, 1240:95-100 (1995); Razinkov et
al., J Gen Physiol, 112:409-22 (1998); Shangguan et al.,
Biochemistry, 35:4956-65 (1996); Gunther-Ausborn et al., J Biol
Chem, 270:29279-85 (1995)). However, LPC's viability as a drug
candidate is questionable since it exerts its effect in a highly
reversible manner, requires high molar concentrations (10% or
higher total lipid content), and can be effectively recycled and
metabolized by cells.
[0006] n-docosanol, a 22-carbon saturated alcohol, is also designed
to inhibit host cell entry of a variety of enveloped viruses (Katz
et al., Proc Natl Acad Sci USA, 88:10825-9 (1991)). However,
n-docosanol appears to inhibit virus-cell fusion by perturbing the
properties of the target cell rather than the virus (Katz et al.,
Ann NY Acad Sci, 724:472-88 (1994); Marcelletti et al., AIDS Res
Hum Retroviruses, 12:71-4 (1996); Pope et al., Antiviral Res,
40:85-94 (1998)), as optimal inhibition is observed when cells, but
not virus, are pre-incubated for several hours with n-docosanol. In
addition, poor solubility and a millimolar IC.sub.50 has limited
n-docosanol to use as a 10% v/v topical microbicide (Abreva.TM.)
for the treatment of cold sores (Katz et al., Ann NY Acad Sci,
724:472-88 (1994); Marcelletti et al., AIDS Res Hum Retroviruses,
12:71-4 (1996); Pope et al., Antiviral Res, 40:85-94 (1998)).
[0007] Recently, amphipathic peptides derived from the NS5A protein
of Hepatitis C have been identified as having antiviral activity
related to their ability to disrupt the membrane integrity of
enveloped viruses (Bobardt et al., Proc Natl Acad Sci USA,
105:5525-30 (2008); Cheng et al., Proc Natl Acad Sci USA,
105:3088-93 (2008)). Although the NS5A-derived peptides were not
active against all enveloped viruses, their broad range of activity
validates viral membranes as a therapeutic target for broad
spectrum antiviral therapeutics.
[0008] Thus, there is a need in the art for broad-spectrum
antiviral drugs capable of disrupting viral lipid membranes and
thereby preventing viral infections.
[0009] Toxoplasma gondii is an obligate intracellular parasite in
the phylum Apicomplexa that causes severe central nervous system
disorders of immunocompromised (AIDS/transplant/lymphoma)
individuals, birth defects in congenitally infected neonates, and
ocular disease in immunocompetent persons. In addition to being an
important pathogen in its own right, Toxoplasma serves as a model
system for studying apicomplexan parasites which cause a number of
diseases of medical and veterinary importance worldwide. There are
over 5000 different species of apicomplexans, but the most notable
of these is Plasmodium falciparum, the causative agent of malaria
which kills 1-2 million people each year. Other Apicomplexans which
cause disease in humans include the opportunistic pathogens
Cryptosporidia spp. and Isospora belli. Important veterinary
pathogens include Neospora caninum (a pathogen of dogs/cattle),
Theileria spp (cattle), and Eimeria spp (poultry).
[0010] Apicomplexan parasites share a common mechanism of gliding
motility that drives invasion into the host cell, a process that is
essential for survival. The parasites secrete an array of molecular
adhesins from a specialized secretory organelle named the
micronemes onto the surface of the parasite which mediate motility
and attachment to the host cell. The cytoplasmic tails of these
adhesins interact with a parasite actin:myosin motor which is
anchored in a membrane system underlying the plasma membrane known
as the inner membrane complex. Action on the actin:myosin motor
provides the driving force for motility, and also for the parasites
to actively penetrate their respective host cells. In Toxoplasma
and Plasmodium, the micronemal protein AMA1 also has been shown to
organize the moving junction RON proteins which are secreted from
another organelle called the rhoptries and this complex of proteins
is critical for subsequent invasion. Current antiparasite therapies
target biosynthetic pathways in the parasite and the bacterial-like
apicoplast and are prone to mutations leading to resistance.
[0011] Thus, there is a need in the art for broad-spectrum
antiparasitic drugs that are less susceptible to resistance through
mutation.
SUMMARY
[0012] In a first embodiment, a compound is provided according to
formula Ia or a pharmaceutically acceptable salt thereof,
##STR00001##
[0013] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.6 aryl, C.sub.3-C.sub.6 heteroaryl, C.sub.3-C.sub.6
cycloalkyl, or C.sub.3-C.sub.6 heterocycloalkyl, each optionally
substituted with halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3,
--SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3,
--OC(O)R.sup.3, --NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0014] R.sup.2 represents one or more substituents independently
selected from --OR.sup.3, --SR.sup.3, and --NR.sup.4R.sup.5;
[0015] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5; and
[0016] X is O, N, S, NH, or NR.sup.3.
[0017] In a second embodiment, a compound is provided according to
formula IIa or a pharmaceutically acceptable salt thereof,
##STR00002##
[0018] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, each optionally substituted with
halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0019] R.sup.2 represents exactly one --OR.sup.3, --SR.sup.3, or
--NR.sup.4R.sup.5;
[0020] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, or C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0021] X is O, N, S, NH, or NR.sup.3; and
[0022] Z is O, N, NH, or NR.sup.3.
[0023] In one aspect of the first or second embodiment, R.sup.3,
R.sup.4, and R.sup.5 are independently C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl.
[0024] In one aspect of the first or second embodiment, R.sup.1 is
C.sub.1-C.sub.6 alkyl.
[0025] In a third embodiment, a method of treating or preventing a
disease or condition caused by infection with an enveloped virus is
provided, the method comprising administering to a subject a
therapeutically effective amount of a compound of formula Ia or
formula IIa.
[0026] In a first aspect of the third embodiment, a method of
treating or preventing a disease or condition caused by infection
with an enveloped virus is provided, the method comprising
administering to a subject a therapeutically effective amount of a
compound according to formula IIa or a pharmaceutically acceptable
salt thereof,
##STR00003##
[0027] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, or C.sub.2-C.sub.6 alkynyl, each optionally substituted
with halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0028] R.sup.2 represents one or more substituents independently
selected from H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2,
--CF.sub.3, --CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3,
--NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0029] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0030] X is O, N, S, NH, or NR.sup.3; and
[0031] Z is O, N, NH, or NR.sup.3.
[0032] In a second aspect of the third embodiment, the virus is
hepatitis C virus (HCV), human immunodeficiency virus (HIV), herpes
simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2),
Ebola virus, Influenza virus, Nipah virus, Yellow Fever Virus,
Dengue virus, Rift Valley Fever Virus, West Nile Virus.
[0033] In a third aspect of the third embodiment, the compound is
administered as a liposomal formulation.
[0034] In a fourth embodiment, a pharmaceutical composition
comprising a compound of formula Ia or formula IIa and at least one
pharmaceutically acceptable excipient is provided. In a first
aspect of the fourth embodiment, the composition is suitable as a
topical formulation. In a second aspect of the fourth embodiment,
the composition is in the form of a patch, an ointment, a cream, a
lotion, a drop, a spray, or an aerosol. In a third aspect of the
fourth embodiment, the composition is in the form of a liposomal
formulation.
[0035] In a fifth embodiment, a method of treating or preventing a
disease or condition caused by infection with a parasite is
provided, the method comprising administering to a subject a
therapeutically effective amount of the compound for formula Ia or
formula IIa.
[0036] In a sixth embodiment, a method of treating or preventing a
disease or condition caused by infection with a parasite is
provided, the method comprising administering to a subject a
therapeutically effective amount of a compound according to formula
IIa or a pharmaceutically acceptable salt thereof,
##STR00004##
[0037] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.6 aryl, C.sub.3-C.sub.6 heteroaryl, C.sub.3-C.sub.6
cycloalkyl, or C.sub.3-C.sub.6 heterocycloalkyl, each optionally
substituted with halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3,
--SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3,
--OC(O)R.sup.3, --NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0038] R.sup.2 represents one or more substituents independently
selected from --OR.sup.3, --SR.sup.3, and --NR.sup.4R.sup.5;
[0039] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0040] X is O, N, S, NH, or NR.sup.3; and
[0041] Z is O, N, NH, or NR.sup.3.
BRIEF DESCRIPTION OF DRAWINGS
[0042] FIG. 1 shows compounds-mediated inhibition of NDV infection.
Virus and compounds, at the indicated concentration, were incubated
under light for 10 minutes and used to infect Vero cells. 24 hours
later, GFP expression in cells was analyzed by flow cytometry.
n=number of independent experiments pooled. Inhibitory
concentration 50% (IC50 and 95% confidence interval (CI95%) are
indicated.
[0043] FIG. 2 shows increased cytotoxicity of the compounds under
light exposure. Compounds at the indicated concentrations were
incubated with Hela cells and left in the dark (0 hour, dashed
lines) or exposed to light for 1 hour (solid lines). 24 hours
later, cell survival was assessed by MTT assay. Inhibitory
concentration 50% (IC50) and 95% confidence interval (CI95%) are
indicated.
[0044] FIG. 3 shows that alpha-tocopherol blocks the antiviral
activity ofJL103 (compound 2). NDV virus encoding GFP was incubated
with or without JL103 in the presence of increasing concentrations
of alpha-tocopherol. Vero cells were then infected with the virus.
At 24 h post-infection, cells were harvested and GFP expression was
measured by flow cytometry.
[0045] FIG. 4 shows HIV inactivation by several compounds of the
present invention in packed red blood cells (RBC) at a hematocrit
of 80%. Packed RBC spiked with HIV (IIIB) were treated for 1 h with
100 .mu.M of the indicated compound. After treatment, the serum was
used to infect the reporter cells TZM-BL. At 48 h after infection,
luciferase expression was measured in cell lysates (RLU).
[0046] FIG. 5 shows the normalized absorption spectra of selected
compounds in DMSO. The absorption spectra of 100 .mu.M solutions of
selected compounds in DMSO were scanned using a Tecan Infinite.RTM.
M1000 microplate reader and normalized to their absorption
maxima.
DETAILED DESCRIPTION
[0047] Provided herein are a series of oxazolidine-2,4-dithiones
having broad-spectrum antiviral activity. These compounds are novel
photosensitizers that act as broad-spectrum, virucidal antivirals
that prevent infection of a variety of viruses, especially lipid
bilayer encapsulated (enveloped) viruses. Without being limited to
a particular theory, it is believed that the compounds are capable
of binding and/or inserting into the outer membranes of enveloped
viruses and disrupting the membrane structure and/or dynamics in a
manner that inhibits viral attachment, fusion, and/or entry into
host cells. Moreover, the membrane disrupting activity of the
compounds is preferably selective for viral membranes relative to
non-viral biological membranes, including but not limited to, cell
plasma membranes, organelle membranes, biological barriers, and
other non-viral lipid bilayers. Thus, compounds provided herein are
substantially non-toxic and have a wide therapeutic index for
antiviral activity in vivo without significant toxicity or side
effects. Because membranes are common to all enveloped viruses,
compounds provided herein have broad ranging antiviral activity
against enveloped viruses, including but not limited to,
filoviruses, poxviruses, arenaviruses, bunyaviruses,
paramyxoviruses, flaviviruses, influenza A, and HIV-1.
[0048] In some aspects, antiviral compounds of formula 1, and
pharmaceutically acceptable salts, prodrugs, and derivatives
thereof, are provided herein
##STR00005##
[0049] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.6 aryl, C.sub.3-C.sub.6 heteroaryl, C.sub.3-C.sub.6
cycloalkyl, or C.sub.3-C.sub.6 heterocycloalkyl, each optionally
substituted with halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3,
--SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3,
--OC(O)R.sup.3, --NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0050] R.sup.2 represents one or more substituents to the ring to
which it is attached, independently selected from H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5,
--SO.sub.2NR.sup.4R.sup.5, aryl, or heteroaryl;
[0051] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0052] Cyc is aryl or heteroaryl;
[0053] X is O, N, S, NH, or NR.sup.3; and
[0054] Z is O, N, NH, or NR.sup.3.
[0055] In some preferred aspects of formula I, Z is O or NH.
[0056] In some aspects, antiviral compounds of formula Ia, and
pharmaceutically acceptable salts, prodrugs, and derivatives
thereof, are provided herein
##STR00006##
[0057] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.6 aryl, C.sub.3-C.sub.6 heteroaryl, C.sub.3-C.sub.6
cycloalkyl, or C.sub.3-C.sub.6 heterocycloalkyl, each optionally
substituted with halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3,
--SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3,
--OC(O)R.sup.3, --NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0058] R.sup.2 represents one or more substituents to the ring to
which it is attached, independently selected from H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0059] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3,
--OC(O)R.sup.3, --NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
and
[0060] X is O, N, S, NH, or NR.sup.3.
[0061] In some preferred aspects of formulas I and Ia, X is O or
N--CH.sub.3.
[0062] In some preferred embodiments of formula I and Ia, R.sup.1
is other than H, such as C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, or C.sub.2-C.sub.6 alkynyl, most preferably methyl, ethyl,
propyl or propenyl.
[0063] In some preferred embodiments of formulas I and Ia, each
R.sup.2 is independently --OR.sup.3, --SR.sup.3, or
--NR.sup.4R.sup.5. In other preferred embodiments of formula Ia,
each R.sup.2 is, independently, --O--C.sub.1-C.sub.6 alkyl,
--O--C.sub.2-C.sub.6 alkenyl, --O--C.sub.2-C.sub.6 alkynyl,
--S--C.sub.1-C.sub.6 alkyl, --S--C.sub.2-C.sub.6 alkenyl,
--S--C.sub.2-C.sub.6 alkynyl or --N(C.sub.1-C.sub.6
alkyl)(C.sub.1-C.sub.6 alkyl). In yet other preferred embodiments,
each R.sup.2 is independently preferably --OMe, --OEt, --OPr,
--SMe, --SEt, --SPr, --N(Me)(Me), --N(Et)(Et) or --N(Pr)(Pr), and
most preferably --OMe or --N(Me)(Me). In some preferred embodiments
of formulas I and Ia, more than one R.sup.2 is present. In some
such embodiments, all R.sup.2 substituents are identical. In other
aspects, exactly one R.sup.2 is present.
[0064] In some preferred embodiments of formula Ia:
[0065] R.sup.1 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
or C.sub.2-C.sub.6 alkynyl;
[0066] X is O or S; and
[0067] one to three R.sup.2 substituents are present, each
independently selected from --O--C.sub.1-C.sub.6 alkyl,
--O--C.sub.2-C.sub.6 alkenyl, --O--C.sub.2-C.sub.6 alkynyl,
--S--C.sub.1-C.sub.6 alkyl, --S--C.sub.2-C.sub.6 alkenyl,
--S--C.sub.2-C.sub.6 alkynyl, and --N(C.sub.1-C.sub.6
alkyl)(C.sub.1-C.sub.6 alkyl).
[0068] In some preferred embodiments of formula Ia:
[0069] R.sup.1 is C.sub.1-C.sub.3 alkyl;
[0070] X is O or S; and
[0071] one to three R.sup.2 substituents are present, each
independently selected from --O--C.sub.1-C.sub.3 alkyl,
--S--C.sub.1-C.sub.3 alkyl, and --N(C.sub.1-C.sub.3
alkyl)(C.sub.1-C.sub.3 alkyl).
[0072] In yet other preferred embodiments of formula Ia, R.sup.1 is
C.sub.1-C.sub.3 alkyl, X is O, Z is O, and two R.sup.2 substituents
are present, each independently --O--C.sub.1-C.sub.3 alkyl.
[0073] Also provided herein are antiviral compounds of formula II,
and pharmaceutically acceptable salts, prodrugs, and derivatives
thereof, are provided herein
##STR00007##
[0074] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.6 aryl, C.sub.3-C.sub.6 heteroaryl, C.sub.3-C.sub.6
cycloalkyl, or C.sub.3-C.sub.6 heterocycloalkyl, each optionally
substituted with halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3,
--SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3,
--OC(O)R.sup.3, --NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0075] R.sup.2 represents one or more substituents to the ring to
which it is attached, independently selected from H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5,
--SO.sub.2NR.sup.4R.sup.5, aryl, or heteroaryl;
[0076] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0077] Cyc is aryl or hetcroaryl;
[0078] X is O, N, S, NH, or NR.sup.3; and
[0079] Z is O, N, NH, or NR.sup.3.
[0080] Also provided herein are antiviral compounds of formula IIa,
and pharmaceutically acceptable salts, prodrugs, and derivatives
thereof, are provided herein
##STR00008##
[0081] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.6 aryl, C.sub.3-C.sub.6 heteroaryl, C.sub.3-C.sub.6
cycloalkyl, or C.sub.3-C.sub.6 heterocycloalkyl, each optionally
substituted with halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3,
--SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3,
--OC(O)R.sup.3, --NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0082] R.sup.2 represents one or more substituents to the ring to
which it is attached, independently selected from H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0083] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0084] X is O, N, S, NH, or NR.sup.3; and
[0085] Z is O, N, NH, or NR.sup.3.
[0086] In some preferred aspects of formula IIa, X is O.
[0087] In some preferred aspects of formulas II and IIa, Z is O. In
other preferred aspects of formulas II and IIa, Z is NH.
[0088] In some preferred embodiments of formulas II and IIa,
R.sup.1 is other than H, such as methyl, ethyl or propyl,
preferably ethyl. In other embodiments, R.sup.1 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl.
[0089] In some preferred embodiments of formulas II and IIa, each
R.sup.2 is H. In other preferred embodiments, of formula IIa, each
R.sup.2 is independently --OR.sup.3, --SR.sup.3, or
--NR.sup.4R.sup.5. In some such embodiments, all R.sup.2
substituents are identical.
[0090] In some aspects, the antiviral compounds of formulas I and
Ia are:
##STR00009## ##STR00010##
[0091]
(Z)-3-ethyl-5-((5-phenylfuran-2-yl)methylene)oxazolidine-2,4-dithio-
ne (i.e., JL102); or
(Z)-3-ethyl-5-((5-(2-methoxyphenyl)furan-2-yl)methylene)oxazolidine-2,4-d-
ithione (i.e., JL103).
[0092] In further aspects, the antiviral compound of formulas IIa
is:
##STR00011##
[0093]
(Z)-3-ethyl-5-((5-phenylfuran-2-yl)methylene)-2-thioxooxazolidine-4-
-one (i.e., JL101).
[0094] In some aspects, antiparasitic compounds of formula III, and
pharmaceutically acceptable salts, prodrugs, and derivatives
thereof, are provided herein
##STR00012##
[0095] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.6 aryl, C.sub.3-C.sub.6 heteroaryl, C.sub.3-C.sub.6
cycloalkyl, or C.sub.3-C.sub.6 heterocycloalkyl, each optionally
substituted with halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3,
--SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3,
--OC(O)R.sup.3, --NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0096] R.sup.2 represents one or more substituents to the ring to
which it is attached, independently selected from H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5,
--SO.sub.2NR.sup.4R.sup.5, aryl, or heteroaryl;
[0097] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0098] Cyc is aryl or heteroaryl;
[0099] X is O, N, S, NH, or NR.sup.3; and
[0100] Z is O, N, NH, or NR.sup.3.
[0101] In some preferred aspects of formula III, Z is O or NH.
[0102] In some aspects, antiparasitic compounds of formula IIIa,
and pharmaceutically acceptable salts, prodrugs, and derivatives
thereof, are provided herein
##STR00013##
[0103] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.6 aryl, C.sub.3-C.sub.6 heteroaryl, C.sub.3-C.sub.6
cycloalkyl, or C.sub.3-C.sub.6 heterocycloalkyl, each optionally
substituted with halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3,
--SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3,
--OC(O)R.sup.3, --NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0104] R.sup.2 represents one or more substituents to the ring to
which it is attached, independently selected from H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0105] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5; and
[0106] X is O, N, S, NH, or NR.sup.3.
[0107] In sonic preferred aspects of formula IIIa, X is O or
N--CH.sub.3.
[0108] In some preferred embodiments of formulas III and IIIa,
R.sup.1 is other than H, such as C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl, most
preferably methyl, ethyl, propyl or propenyl. In other embodiments,
R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl.
[0109] In some preferred embodiments of formulas III and IIIa, each
R.sup.2 is independently --OR.sup.3, --SR.sup.3, or
--NR.sup.4R.sup.5. In other preferred embodiments of formulas IIIa,
each R.sup.2 is, independently, --O--C.sub.1-C.sub.6 alkyl,
--O--C.sub.2-C.sub.6 alkenyl, --O--C.sub.2-C.sub.6 alkynyl,
--S--C.sub.1-C.sub.6 alkyl, --S--C.sub.2-C.sub.6 alkenyl,
--S--C.sub.2-C.sub.6 alkynyl or --N(C.sub.1-C.sub.6
alkyl)(C.sub.1-C.sub.6 alkyl). In other preferred embodiments, each
R.sup.2 is independently preferably --OMe, --OEt, --OPr, --SMe,
--SEt, --SPr, --N(Me)(Me), --N(Et)(Et) or --N(Pr)(Pr), and most
preferably --OMe or --N(Me)(Me). In some preferred embodiments of
formulas III and IIIa, more than one R.sup.2 is present. In some
such embodiments, all R.sup.2 substituents are identical. In other
aspects, exactly one R.sup.2 is present.
[0110] In some preferred embodiments of formula IIIa:
[0111] R.sup.1 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
or C.sub.2-C.sub.6 alkynyl;
[0112] X is O or S; and
[0113] one to three R.sup.2 substituents are present, each
independently selected from --O--C.sub.1-C.sub.6 alkyl,
--O--C.sub.2-C.sub.6 alkenyl, --O--C.sub.2-C.sub.6 alkynyl,
--S--C.sub.1-C.sub.6 alkyl, --S--C.sub.2-C.sub.6 alkenyl,
--S--C.sub.2-C.sub.6 alkynyl, and --N(C.sub.1-C.sub.6
alkyl)(C.sub.1-C.sub.6 alkyl).
[0114] In some preferred embodiments of formula IIIa:
[0115] R.sup.1 is C.sub.1-C.sub.3 alkyl;
[0116] X is O or S; and
[0117] one to three R.sup.2 substituents are present, each
independently selected from --O--C.sub.1-C.sub.3 alkyl,
--S--C.sub.1-C.sub.3 alkyl, and --N(C.sub.1-C.sub.3
alkyl)(C.sub.1-C.sub.3 alkyl).
[0118] In yet other preferred embodiments of formula IIIa, R.sup.1
is C.sub.1-C.sub.3 alkyl, X is O; and two R.sup.2 substituents are
present, each independently --O--C.sub.1-C.sub.3 alkyl.
[0119] Also provided herein are antiparasitic compounds of formula
IV, and pharmaceutically acceptable salts, prodrugs, and
derivatives thereof, are provided herein
##STR00014##
[0120] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.6 aryl, C.sub.3-C.sub.6 heteroaryl, C.sub.3-C.sub.6
cycloalkyl, or C.sub.3-C.sub.6 heterocycloalkyl, each optionally
substituted with halo, --NO.sub.2, --CF.sub.3, --CN, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0121] R.sup.2 represents one or more substituents to the ring to
which it is attached, independently selected from H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5,
--SO.sub.2NR.sup.4R.sup.5, aryl, or heteroaryl;
[0122] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0123] Cyc is aryl or heteroaryl;
[0124] X is O, N, S, NH, or NR.sup.3; and
[0125] Z is O, N, NH, or NR.sup.3.
[0126] Also provided herein are antiparasitic compounds of formula
IVa, and pharmaceutically acceptable salts, prodrugs, and
derivatives thereof, are provided herein
##STR00015##
[0127] wherein R.sup.1 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.6 aryl, C.sub.3-C.sub.6 heteroaryl, C.sub.3-C.sub.6
cycloalkyl, or C.sub.3-C.sub.6 heterocycloalkyl, each optionally
substituted with halo, --NO.sub.2, --CF.sub.3, --CN, --OR.sup.3,
--SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3,
--OC(O)R.sup.3, --NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5; halo,
--NO.sub.2, --CF.sub.3, --CN, --OR.sup.3, --SR.sup.3,
--C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3, --OC(O)R.sup.3,
--NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0128] R.sup.2 represents one or more substituents to the ring to
which it is attached, independently selected from H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --OR.sup.3, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3,
--C(O)OR.sup.3, --OC(O)R.sup.3, --NR.sup.4R.sup.5,
--C(O)NR.sup.4R.sup.5, --NHR.sup.3C(O)NR.sup.4R.sup.5, or
--SO.sub.2NR.sup.4R.sup.5;
[0129] R.sup.3, R.sup.4, and R.sup.5 are independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxy, halo, --NO.sub.2, --CF.sub.3,
--CN, --SR.sup.3, --C(O)R.sup.3, --NHC(O)R.sup.3, --C(O)OR.sup.3,
--OC(O)R.sup.3, --NR.sup.4R.sup.5, --C(O)NR.sup.4R.sup.5,
--NHR.sup.3C(O)NR.sup.4R.sup.5, or --SO.sub.2NR.sup.4R.sup.5;
[0130] X is O, N, S, NH, or NR.sup.3; and
[0131] Z is O, N, NH, or NR.sup.3.
[0132] In some preferred aspects of formula IVa, X is O.
[0133] In some preferred aspects of formula IVa, Z is O. In other
preferred aspects of formula IVa, Z is NH.
[0134] In some preferred embodiments of formula IVa, R.sup.1 is
other than H, such as C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, in particular methyl, ethyl or
propyl, preferably ethyl. In other embodiments, R.sup.1 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl.
[0135] In some preferred embodiments of formula IVa, each R.sup.2
is H. In other preferred embodiments, of formula IVa, each R.sup.2
is independently --OR.sup.3, --SR.sup.3, or --NR.sup.4R.sup.5. In
some such embodiments, all R.sup.2 substituents are identical.
[0136] In some aspects, the antiparasitic compounds of formulas III
and IIIa are:
##STR00016## ##STR00017##
[0137]
(Z)-3-ethyl-5-((5-phenylfuran-2-yl)methylene)oxazolidine-2,4-dithio-
ne (i.e., JL102); or
(Z)-3-ethyl-5-((5-(2-methoxyphenyl)furan-2-yl)methylene)oxazolidine-2,4-d-
ithione (i.e., JL103).
[0138] In further aspects, the antiparasitic compound of formulas
IVa is:
##STR00018##
Or
[0139]
(Z)-3-ethyl-5-((5-phenylfuran-2-yl)methylene)-2-thioxooxazolidine-4-
-one (i.e., JL101).
[0140] In some aspects, the following antiviral compounds, and
pharmaceutically acceptable salts, prodrugs, and derivatives
thereof, are provided herein:
##STR00019##
[0141] In some aspects, the following antiparasitic compounds, and
pharmaceutically acceptable salts, prodrugs, and derivatives
thereof, are provided herein:
##STR00020##
[0142] Chemical moieties referred to as univalent chemical moieties
(e.g., alkyl, aryl, etc.) also encompass structurally permissible
multivalent moieties, as understood by those skilled in the art.
For example, while an "alkyl" moiety generally refers to a
monovalent radical (e.g., CH.sub.3CH.sub.2--), in appropriate
circumstances an "alkyl" moiety can also refer to a divalent
radical (e.g., --CH.sub.2CH.sub.2--, which is equivalent to an
"alkylene" group). Similarly, under circumstances where a divalent
moiety is required, those skilled in the art will understand that
the term "aryl" refers to the corresponding divalent arylene
group.
[0143] All atoms are understood to have their normal number of
valences for bond formation (e.g., 4 for carbon, 3 for N, 2 for O,
and 2, 4, or 6 for S, depending on the atom's oxidation state). On
occasion a moiety can be defined, for example, as (A).sub.aB,
wherein a is 0 or 1. In such instances, when a is 0 the moiety is B
and when a is 1 the moiety is AB.
[0144] Where a substituent can vary in the number of atoms or
groups of the same kind (e.g., alkyl groups can be C.sub.1,
C.sub.2, C.sub.3, etc.), the number of repeated atoms or groups can
be represented by a range (e.g., C.sub.1-C.sub.6 alkyl) which
includes each and every number in the range and any and all sub
ranges. For example, C.sub.1-C.sub.3 alkyl includes C.sub.1,
C.sub.2, C.sub.3, C.sub.1-2, C.sub.1-3, and C.sub.2-3 alkyl.
[0145] The terms "alkyl," "alkenyl," and "alkynyl," refer to
straight and branched chain aliphatic groups having from 1 to 30
carbon atoms, or preferably from 1 to 15 carbon atoms, or more
preferably from 1 to 6 carbon atoms, each optionally substituted
with one, two or three substituents depending on valency. Examples
of such groups include, without limitation, methyl, ethyl, propyl,
isopropyl, butyl, tent-butyl, isobutyl, pentyl, hexyl, vinyl,
allyl, isobutenyl, ethynyl, and propynyl.
[0146] The term "cycloalkyl" includes saturated and partially
unsaturated cyclic hydrocarbon groups having from 3 to 12, or
preferably from 3 to 8, or more preferably from 3 to 6 carbon
atoms, each optionally substituted with one or more substituents.
Examples of cycloalkyl groups include, without limitation,
cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl,
cyclohexenyl, cycloheptyl, and cyclooctyl.
[0147] An "aryl" group is an optionally substituted
C.sub.6-C.sub.14 moiety comprising one to three aromatic rings. In
some aspects, the aryl group is a C.sub.6-C.sub.10 aryl group, or
more preferably a C.sub.5-C.sub.6 aryl group. Examples of aryl
groups include, without limitation, phenyl, naphthyl, anthracenyl,
and fluorenyl.
[0148] A "heterocyclic" or "heterocyclyl" substituent is a
non-aromatic mono-, bi-, or tricyclic structure having from about 3
to about 14 atoms, including one or more heteroatoms selected from
N, O, and S. One ring of a bicyclic heterocycle or two rings of a
tricyclic heterocycle can be aromatic (e.g., as in indan and
9,10-dihydro anthracene). Heterocyclic groups can be optionally
substituted on one or more carbon, oxygen, nitrogen and/or sulfur
atoms. Examples of heterocyclic groups include, without limitation,
epoxy, aziridinyl, tetrahydrofuranyl, pyrrolidinyl, piperidinyl,
piperazinyl, thiazolidinyl, oxazolidinyl, oxazolidinonyl, and
morpholino.
[0149] A "heteroaryl" group is an aromatic ring or ring system
having about 5 to 14 ring atoms, or more preferably 5, 6, 9, or 10
ring atoms, including one or more heteroatoms selected from the
group consisting of N, O, and S; and 6, 10, or 14 pi electrons
shared in a cyclic array. Examples of heteroaryl groups include,
without limitation, thienyl, benzothienyl, furyl, benzofuryl,
dibenzofuryl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl,
pyrimidinyl, indolyl, quinolyl, isoquinolyl, quinoxalinyl,
tetrazolyl, oxazolyl, thiazolyl, and isoxazolyl.
[0150] A "substituted" moiety is a moiety in which one or more
hydrogen atoms have been independently replaced with another
chemical substituent. As a non limiting example, substituted phenyl
groups include 2-fluorophenyl, 3,4-dichlorophenyl,
3-chloro-4-fluorophenyl, and 2-fluoro-3-propylphenyl. In some
instances, a methylene group (--CH.sub.2--) is substituted with
oxygen to form a carbonyl group (--CO).
[0151] An "optionally substituted" group can be substituted with
from one to four, or preferably from one to three, or more
preferably one or two non-hydrogen substituents. Examples of
suitable substituents include, without limitation, alkyl, alkenyl,
alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aroyl,
halo, hydroxy, oxo, nitro, alkoxy, amino, imino, azido, mercapto,
acyl, carbamoyl, carboxy, carboxamido, amidino, guanidino,
sulfonyl, sulfinyl, sulfonamido, formyl, cyano, and ureido
groups.
[0152] The term "halogen" or "halo" refers to chlorine, bromine,
fluorine, or iodine.
[0153] The term "acyl" refers to an alkylcarbonyl or arylcarbonyl
substituent.
[0154] The term "acylamino" refers to an amide substituent attached
to the structure at the nitrogen atom. Acylamino groups can be
optionally substituted.
[0155] The term "carbamoyl" refers to an amide substituent attached
to the structure at the carbonyl carbon atom. Carbamoyl groups can
be optionally substituted.
[0156] The term "sulfonamido" refers to a sulfonamide substituent
attached to the structure by either the sulfur or the nitrogen
atom.
[0157] Unless otherwise specified, compounds provided herein
include all of their various stereochemical forms, including but
not limited to, enantiomers, diastereomers, rotamers, and the like.
Also, moieties disclosed herein which exist in multiple tautomeric
forms include all such forms encompassed by a given tautomeric
structure.
[0158] Particular geometric isomers (e.g., E or Z isomers)
disclosed herein include the E or Z isomer substantially free from
the other isomer as well as mixtures of E and Z isomers in varying
ratios. For example, in some preferred aspects, compounds provided
herein comprise the (Z)-isomer substantially free from the
(E)-isomer.
[0159] Certain E and Z geometric isomers can be interconverted by
photolysis, photo irradiation or exposure to free radicals or
certain solvents (sec e.g., lshida et al., Tetrahedron Lett 30:959
(1989)). For example, exposure of some (E) compounds to DMSO
facilitates their conversion to the Z form.
[0160] Compounds provided herein can form useful salts with
inorganic and organic acids, such as hydrochloric, sulfuric,
acetic, lactic, or the like, and with inorganic or organic bases
such as sodium or potassium hydroxide, piperidine, morpholine,
ammonium hydroxide, or the like. Pharmaceutically acceptable salts
of compounds provided herein can be prepared using procedures
familiar to those skilled in the art.
[0161] Also provided herein are pharmaceutical compositions
comprising an antiviral compound described herein and at least one
pharmaceutically acceptable excipient.
[0162] The term "pharmaceutically acceptable excipient" includes
any and all salts, buffering agents, preservatives, solvents,
diluents, carriers, liquid vehicles, dispersion or suspension aids,
surface active agents, isotonic agents, thickening or emulsifying
agents, preservatives, solid binders, lubricants and the like which
are compatible with the antiviral compounds provided herein and
suitable for the particular dosage form desired. Various carriers,
formulations, and techniques are described, e.g., in Remington's
Pharmaceutical Sciences, E. W. Martin (Mack Publishing Co., Easton,
Pa.).
[0163] Pharmaceutical compositions provided herein are formulated
to be compatible with their intended route of administration.
Exemplary routes of administration include, e.g., parenteral,
intravenous, intramuscular, intradermal, subcutaneous, oral (e.g.,
inhalation), transdermal (topical), transmucosal, genital, vaginal,
cervicovaginal and rectal administration.
[0164] Solutions or suspensions used for parenteral, intradermal,
or subcutaneous application can include, e.g., a sterile diluent
such as water for injection, saline solution, fixed oils,
polyethylene glycols, glycerine, propylene glycol or other
synthetic solvents; antibacterial agents such as benzyl alcohol or
methyl parabens; antioxidants such as ascorbic acid or sodium
bisulfite; chelating agents such as ethylenediaminctetraacetic
acid; buffers such as acetates, citrates or phosphates and agents
for the adjustment of tonicity such as sodium chloride or dextrose.
The parenteral preparation can be enclosed in ampoules, disposable
syringes or multiple dose vials made of glass or plastic.
[0165] In some aspects, compounds and compositions provided herein
are administered topically, e.g., by transmucosal or transdermal
means. Suitable formulations for topical administration, including,
e.g., vaginal or rectal administration, include solutions,
suspensions, gels, lotions and creams as well as discrete units
such as suppositories and microencapsulated suspensions. For
transmucosal or transdermal administration, penetrants appropriate
to the barrier to be permeated can be used. Such penetrants are
generally known in the art, and include, for example, detergents,
bile salts, and filsidic acid derivatives for transmucosal
administration. Transmucosal administration can be accomplished
through the use of nasal sprays, suppositories or transdermal
formulations comprising active compounds formulated with ointments,
salves, gels, creams, or the like.
[0166] In further aspects, compositions provided herein can be
formulated as tablets, capsules or elixirs for oral administration
or as sterile solutions or suspensions for injectable
administration.
[0167] In some aspects, a compounds described herein is formulated
as a sustained release composition which provides for slow,
sustained release of the compound by a desired mode of
administration. Such formulations can take the form of a sustained
release gel, cream, suppository, capsule, or the like. In some
aspects, active compounds are formulated within a system of
carriers and excipients that protect the compound against rapid
elimination from the body. Examples of such sustained release
systems include: (a) erosional systems in which the active compound
is contained within a matrix, and (b) diffusional systems in which
the active component permeates at a controlled rate through a
biocompatible polymer, such as ethylene vinyl acetate,
polyanhydrides, polyglycolic acid, collagen, polyorthoesters, or
polylactic acid. Methods for preparation of such formulations are
known to those skilled in the art.
[0168] Pharmaceutical compositions provided herein can be utilized
in conjunction with a delivery device, such as a condom or other
contraceptive device, a metered dose inhaler, a transdermal patch,
an implantable pump, sponge, or other reservoir, or the like.
[0169] In some aspects, pharmaceutical compositions provided herein
can be delivered via an intranasal spray, by inhalation, and/or by
an aerosol. Methods for delivering pharmaceutical compositions
directly to the lungs and/or nasal mucosa via nasal and/or
pulmonary aerosols are well-known in the pharmaceutical arts. In
further aspects, pharmaceutical compositions provided herein can be
delivered ocularly, e.g., via eyedrops.
[0170] In some preferred aspects, pharmaceutical compositions
provided herein are delivered via a liposomal nanoparticle
formulation. For example, in some aspects, the compounds can be
formulated within liposomes comprising a lipid bilayer formulated
to enhance solubility and/or permeability across viral membranes.
In further aspects, a liposomal nanoparticle formulation provided
herein can comprise liposomes having a size range which facilitates
delivery of the active compounds to viral membranes. Liposomal
formulations can be prepared according to methods known to those
skilled in the art.
[0171] Pharmaceutical compositions suitable for injectable use
include sterile aqueous solutions (for water soluble compounds) or
dispersions and sterile powders for the extemporaneous preparation
of sterile injectable solutions or dispersions.
[0172] For intravenous administration, suitable carriers include,
e.g., physiological saline, bacteriostatic water, Cremophor EL.TM.
(BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). The
compositions are preferably sterile and fluid to allow for easy
syringability.
[0173] Oral compositions generally include an inert diluent or an
edible carrier, and can be incorporated with excipients in the form
of tablets, troches, capsules, or the like. Oral compositions can
also be prepared using a fluid carrier for use as a mouthwash or
rinse wherein the compound in the fluid carrier is applied orally
and swished and expectorated or swallowed. Tablets, pills,
capsules, troches and the like can further comprise one or more of
the following: binding agents, such as microcrystalline cellulose,
gum tragacanth or gelatin; adjuvants, such as starch or lactose,
disintegrating agents, such as alginic acid, Primogel, or corn
starch; lubricants, such as magnesium stearate or Sterotes;
glidants, such as colloidal silicon dioxide; and sweetening or
flavoring agents, such as sucrose, saccharin, peppermint, methyl
salicylate, or orange flavoring.
[0174] Pharmaceutical compositions provided herein are preferably
stable under the conditions of manufacture and storage and
preserved against the contaminating action of microorganisms such
as bacteria and fungi. The carrier can be a solvent or dispersion
medium containing, for example, water, ethanol, polyol (e.g.,
glycerol, propylene glycol, liquid polyetheylene glycol, or the
like), and suitable mixtures thereof. The proper fluidity can be
maintained, for example, by the use of a coating such as lecithin,
by the maintenance of the required particle size in the case of a
dispersion, and by the use of surfactants. Prevention of the action
of microorganisms can be achieved by various antibacterial and
antifungal agents, including, for example, parabens, chlorobutanol,
phenol, ascorbic acid, thimerosal, and the like.
[0175] In some aspects, pharmaceutical compositions provided herein
are formulated in dosage unit form (physically discrete units
comprising a unitary, predetermined quantity of active compound
calculated to produce the desired therapeutic effect in association
with the required pharmaceutical carrier). The exact specifications
of dosage unit forms will be dictated by the unique characteristics
of the active compound, the particular therapeutic effect to be
achieved, the preferred route of administration, and the like.
[0176] Toxicity and therapeutic efficacy antiviral compounds
provided herein can be determined using standard pharmaceutical
procedures in cell cultures or experimental animals. For example,
established methods can be used to calculate LD.sub.50 (the dose
lethal to 50% of the population) and/or ED.sub.50 (the dose
therapeutically effective in 50% of the population) doses for the
antiviral compounds. The dose ratio between toxic and therapeutic
effects is the therapeutic index, which can be expressed as the
ratio LD.sub.50/ED.sub.50. Compounds which exhibit large
therapeutic indices are generally preferred, as are formulations
and modes of administration which enhance the therapeutic index for
a particular compound.
[0177] Data obtained from the cell culture assays and animal
studies can be used in formulating a range of dosage for use in
humans. In some aspects, dosages of antiviral compounds provided
herein lie within a range of circulating concentrations which
include the ED.sub.50 with little or no toxicity. Dosages can vary
within this range depending upon the dosage form, route of
administration, and the like.
[0178] Therapeutically effective doses of compounds provided herein
can be estimated initially from cell culture assays, e.g., based on
the IC.sub.50 (the concentration of a test compound which achieves
a half-maximal inhibition of viral activity and/or infection). For
example, the IC.sub.50 observed in cell culture assays can be used
to formulate a working dosage range for use in animal models in
order to achieve a circulating plasma concentration range that
includes the IC.sub.50 with little or no toxicity. Conversion
factors and calculation methods for converting animal dosages to
human dose estimates arc well known in the pharmaceutical arts.
Concentrations of free compounds in plasma can be measured, for
example, by high performance liquid chromatography.
[0179] As defined herein, a therapeutically effective amount of an
antiviral compound provided herein (an effective dosage) can range
from about 0.001 to 3000 mg/kg body weight, preferably from about
0.01 to 2500 mg/kg body weight, more preferably about 0.1 to 2000
mg/kg body weight, and even more preferably about 1 to 1000 mg/kg,
5 to 500 mg/kg, 10 to 100 mg/kg body weight. Skilled artisans will
appreciate that a variety of factors can influence the dosage
required to effectively treat a subject, including but not limited
to the severity of the disease or condition being treated, history
of previous treatments, general health and/or age of the subject,
and the like. Accordingly, exact dosages for any particular subject
will typically be determined empirically.
[0180] Pharmaceutical compositions provided herein can comprise a
compound provided herein in an amount of at least 0.5% and
generally not more than 90% by weight, based on the total weight of
the composition, including excipients, if any. In some aspects, the
proportion of antiviral agent varies between about 5-50% by weight
of the composition.
[0181] Also provided herein are methods for treating or preventing
infection by an enveloped virus, comprising administering an
effective amount of a compound described herein to a subject in
need of treatment.
[0182] As used herein, "treating" includes prevention,
amelioration, alleviation, and/or elimination of a disease,
disorder, or condition being treated or one or more symptoms of a
disease, disorder, or condition being treated, as well as
improvement in the overall well being of a patient, as measured by
objective and/or subjective criteria.
[0183] In some aspects, the subject has been infected or is at risk
of infection by an enveloped virus. A subject at risk of an
enveloped virus infection can include any subject that has been
exposed to or is likely to become exposed to an enveloped virus
(e.g., via the skin or mucosal membranes). For example, subjects at
risk can include medical providers, hospital staff and family
members having contact with infected patients, as well as
laboratory or quarantine facility workers having contact with
samples, tissues, and like from infected patients.
[0184] In some aspects, the antiviral activity of a compound
provided herein within a subject can be measured by assaying viral
replication, viral infectivity, and/or viral load, and/or by
measuring one or more secondary indicators of viral infection, such
as indicators of inflammatory and/or immune responses.
[0185] Compounds and methods provided herein are useful for
treating and preventing infections by any enveloped virus.
"Enveloped" viruses are animal viruses having an outer membrane or
`envelope` comprised of a lipid bilayer with embedded viral
proteins.
[0186] In some aspects, the enveloped virus is a type T Filoviridae
virus which has a single-stranded, unsegmented (-) sense RNA genome
and which causes severe hemorrhagic fever in humans and non-human
primates. In some aspects, the Filoviridae virus is an Ebola virus,
such as a Cote d'lvoire (CI), Sudan (S), Zaire (Z) or Reston (R)
species of Ebola virus. In further aspects, the Filoviridae virus
is a Marburg virus.
[0187] In some aspects, the virus is an Orthomyxoviridae virus,
such as an influenza virus, Thogotovirus, Dhori virus, or
infectious salmon anemia virus. For example, in some aspects,
methods provided herein are used to treat or prevent infection of a
human subject with an influenza type A virus, an influenza type B
virus, or an influenza type C virus. In some aspects, the influenza
type A virus is of subtype H1N1, H2N2, H3N2 or H5N1.
[0188] In some aspects, the virus is a Paramyxoviridae virus, such
as human parainfluenza virus, human respiratory syncytial virus
(RSV), Sendai virus, Newcastle disease virus, mumps virus, rubeola
(measles) virus, Hendra virus, Nipah virus, avian pneumovirus, or
canine distemper virus.
[0189] In some aspects, the virus is a Rhabdoviridae virus, such as
rabies virus, vesicular stomatitis virus (VSV), Mokola virus,
Duvenhage virus, European bat virus, salmon infectious
hematopoietic necrosis virus, viral hemorrhagic septicaemia virus,
spring viremia of carp virus, or snakehead rhabdovirus.
[0190] In some aspects, the virus is a Bornaviridae virus, such as
Borna disease virus.
[0191] In some aspects, the virus is a Bunyaviridae virus, such as
Bunyamwera virus, Hantaan virus, Crimean Congo virus, California
encephalitis virus, Rift Valley fever virus, or sandfly fever
virus.
[0192] In some aspects, the virus is an Arenaviridae virus, such as
Old World Arenaviruses, Lassa fever virus, Ippy virus, Lymphocytic
choriomeningitis virus (LCMV), Mobala virus, Mopeia virus, or a New
World Arenavirus, such as Junin virus (Argentine hemorrhagic
fever), Sabia (Brazilian hemorrhagic fever), Amapari virus, Flexal
virus, Guanarito virus (Venezuela hemorrhagic fever), Machupo virus
(Bolivian hemorrhagic fever), Latino virus, Boliveros virus, Parana
virus, Pichinde virus, Pirital virus, Tacaribe virus, Tamiami
virus, or Whitewater Arroyo virus. In some aspects, the
Arenaviridae virus is Lymphocytic choriomeningitis virus, Lassa
virus, Junin Virus, Machupo Virus, Sabia virus, or Guanarito
virus.
[0193] In some aspects, the virus is an arbovirus. Arboviruses
comprise a large group of more than 400 enveloped RNA viruses that
arc transmitted primarily by arthropod vectors (e.g., mosquitoes,
sand-flies, fleas, ticks, lice, etc). In some aspects, the
arbovirus is a Togaviridae virus, such as an Alphavirus (e.g.,
Venezuela equine encephalitis virus or Sindbis virus) or a
Rubivirus (e.g., Rubella virus). For example, in some aspects, a
compound provided herein is administered to a pregnant subject to
treat or prevent congenital rubella syndrome (CRS) and symptoms
related thereto, such as low birth weight, deafness, and
abortion.
[0194] In some aspects, the arbovirus is a Flaviviridae virus, such
as a Flavivirus, a Pestivirus, a Hepadvirus, yellow fever virus,
dengue fever virus, or Japanese encaphilitis (JE) virus.
[0195] In some aspects, the virus is a Hepacivirus, such as a
hepatitis C virus or a hepatitis C-like virus.
[0196] In some aspects, the virus is a Henipavirus, such as Hendra
virus or Nipah virus.
[0197] In further aspects, the virus is a Bunyaviridae (-)-sense
RNA virus, such as an Orthobunyavirus, a Hantavirus, a Phlebovirus,
or a Nairovirus.
[0198] In some aspects, the virus is a Arenavirius virus, such as
Lymphocytic choriomeningitis virus (LCMV), Lassa virus, Junin
virus, Machupo virus, or Guanarito virus.
[0199] In some aspects, the virus is a Japanese encephalitis virus,
such as Alfuy virus, Japanese encephalitis virus, Kokobera virus,
Koutango virus, Kunjin virus, Murray Valley encephalitis virus, St.
Louis encephalitis virus, Stratford virus, Usutu virus, or West
Nile virus.
[0200] In some aspects, the virus is human immunodeficiency virus
(HIV).
[0201] In some aspects, the virus is a herpesvirus, for example,
HSV-1 or HSV-2.
[0202] Also provided herein are methods of treating a disease or
condition associated with an enveloped virus infection. For
example, in some aspects, methods are provided herein for treating
Ebola Hemorrhagic Fever (EHF), Marburg hemorrhagic fever (MHF),
Dengue fever, Dengue hemorrhagic fever (DHF), yellow fever, dengue
fever, acute and chronic hepatitis C, Venezuelan hemorrhagic fever,
Brazilian hemorrhagic fever, Bolivian hemorrhagic fever,
lymphocytic choriomeningitis, Lassa fever, hantavirus pulmonary
syndrome (HPS), meningitis, influenza, AIDS, and/or genital
herpes.
[0203] In some aspects, a compound provided herein is administered
in combination with an antiviral agent or an antiviral vaccine.
[0204] Compounds and compositions provided herein can be
administered in any amount and via any route of administration
effective for attenuating infectivity of an enveloped virus.
Exemplary routes of administration include, but are not limited to,
oral, intrathecal, intra-arterial, direct bronchial application,
parenteral (e.g., intravenous), intramuscular, intranasal,
sublingual, intratracheal, inhalation, ocular, vaginal, and rectal
administration.
[0205] The term effective amount refers to the amount necessary or
sufficient to realize a desired biologic effect. As understood by
those skilled in the art, an effective amount for any particular
application can vary depending on such factors as the disease or
condition being treated, the particular inhibitor being
administered, the size of the subject, the severity of the disease
or condition being treated, and/or other factors.
[0206] In some aspects, an effective amount of a compound provided
herein is an amount that, when administered via a preferred mode of
administration, is effective to treat or prevent viral infection in
the subject without causing substantial toxicity or adverse
side-effects. In further aspects, an effective amount of an
antiviral compound provided herein is an amount which effectively
inhibits fusion of an enveloped virus with the plasma membrane of a
cell. In additional aspects, an effective amount of an antiviral
compound described herein is an amount which is effective to
ameliorate one or more symptoms associated with an enveloped virus
infection.
[0207] One of ordinary skill in the art can empirically determine
effective amounts of antiviral compounds provided herein by routine
experimentation.
[0208] In some aspects, methods are provided for preventing the
spread of a sexually transmitted disease caused by an enveloped
virus, such as but not limited to a herpesvirus (e.g., HSV-1 or
HSV-2) or HIV, comprising administering an antiviral compound
described herein to a subject who is at risk of being exposed to an
enveloped virus. In some preferred aspects, the antiviral compound
is administered as a topical formulation. In further preferred
aspects, the antiviral compound is administered in conjunction with
a device, such as but not limited to a condom or other
contraceptive device.
[0209] In additional aspects, methods are provided for preventing
infection due to an intentional exposure to an enveloped virus, for
example related to biological warfare or terrorism, wherein the
methods comprise administering an antiviral compound described
herein to a subject who is at risk of being exposed to an enveloped
virus.
[0210] Also provided herein are methods for inactivating enveloped
viruses in a biological or pharmaceutical preparation, the methods
comprising adding an antiviral compound provided herein to the
preparation and incubating the mixture for a time sufficient to
inactivate enveloped viruses present in the preparation.
[0211] In some preferred aspects, the antiviral compounds are
substantially inert with respect to the structure and function of
macromolecules, cells, tissues, organs and/or other biological
structures comprising the preparation. For example, in some
aspects, treating biological preparations with the antiviral
compounds at a concentration and time sufficient to inactivate
enveloped viruses within the preparation does not result in
detectable protein denaturation, protein degradation, plasma
membrane disruption, cell lysis, or the like.
[0212] In additional aspects, biological preparations treated with
the antiviral compounds, at a concentration and time sufficient to
inactivate enveloped viruses within the preparation, are
substantially non-toxic to subjects to whom the preparations are
intended for delivery, including but not limited to, human
subjects. In some preferred aspects, biological preparations
treated with the antiviral compounds are substantially non-toxic to
human subjects without the need for further purification or
processing.
[0213] In some aspects, the biological preparation is a biological
sample drawn from a human or animal donor, such as but not limited
to, blood, plasma, cerebrospinal fluid, mammary fluid, embryonic
fluid, mucus, urine, and the like. For example, in some aspects,
blood, tissue, or an organ harvested from a human or animal donor
is treated according to methods provided herein to inactivate
enveloped viruses, such as but not limited to HIV, prior to
transplantation into a human or animal recipient. In some preferred
aspects, the treated donor sample is transplanted into the
recipient without removing the antiviral compound(s).
[0214] In further aspects, the biological preparation comprises
cultured cells, tissues, or organs, such as but not limited to stem
cells or xerographic tissues intended for transplantation. In
additional aspects, the biological preparation comprises cultured
host cells for the production of a recombinant protein or other
biological product. In some preferred aspects, treating a cellular
preparation with an antiviral compound provided herein inactivates
enveloped viruses within the preparation without substantially
affecting the growth, proliferation, viability, and/or productivity
of the cells. In some preferred aspects, the treated cells are used
or harvested without removing the antiviral compound(s).
[0215] Also provided herein are kits comprising a container housing
an antiviral compound provided herein and instructions for
administering the compound to a subject that has been infected or
is at risk of infection by an enveloped virus. The instructions can
provide for administration as an oral formulation, by inhalation,
by topical administration, by intravenous injection and/or by any
other suitable means.
[0216] In some aspects, kits provided herein optionally further
comprise a pharmaceutical preparation vial and a pharmaceutically
acceptable diluent, such as physiological saline for diluting a
concentrated solution, salt or lyophilized powder formulation of an
antiviral compound provided herein.
[0217] In some aspects, the kit comprises an inhaler for
aerosolized administration to the lungs and/or upper respiratory
tract. In further aspects, the kit comprises a device for
intranasal administration to the nasal mucosa.
[0218] We have discovered that a recently developed series of
pan-enveloped viral inhibitors, including but not limited to LJ001
and compounds of the formula IIIa and IVa, previously shown to
prevent virus-cell membrane fusion and entry, are highly effective
in inhibiting attachment, invasion and motility in Toxoplasma
gondii and are useful as broad-spectrum antiparasitic agents.
Motility and invasion are conserved processes in a wide array of
related apicomplexan parasites, indicating that the compounds will
be effective against this phylum of obligate intracellular
parasites.
[0219] The antiparasitic activity reported herein suggests that the
compounds block the release of parasite proteins from the secretory
micronemes, opening an entirely new approach for controlling
Toxoplasma and related parasites. Downstream events in invasion may
also be affected (e.g., rhoptry release) as these similarly involve
secretory membrane fusion events. Current anti-parasite therapies
target biosynthetic pathways in the parasite and the bacterial-like
apicoplast and are prone to mutations leading to resistance. The
action of blocking membrane fusion processes provides a unique
method of controlling this phylum of obligate intracellular
parasites.
[0220] The processes of motility and invasion in T. gondii and
related apicomplexan parasites require the release of adhesins from
a specialized secretory organelle called the micronemes. As both
motility and attachment are dramatically affected by the present
compounds, the likely mechanism of inhibition is by disrupting
fusion of the micronemes with the parasite plasma membrane and the
subsequent release of adhesins. Additional steps downstream in the
invasion process may also be affected (e.g., secretion from the
rhoptries or dense granules which are other secretory organelles
involved in invasion). The details of inhibition of motility and
attachment are described in more detail herein.
[0221] The specific antiviral activity of the present compounds
involves an ability to interact with the membranes of all
lipid-enveloped viruses tested to date, and to disrupt subsequent
virus-cell membrane fusion. As described herein, we have discovered
that the present compounds also have antiparasitic activity
relating to an ability to inhibit parasite attachment and invasion
processes involving fusion of intracellular organellar membranes
(in the parasite) with the plasma membrane.
[0222] In some aspects, compounds provided herein and derivatives
thereof can be used for therapeutic treatment in humans or animals
of (both acute and chronic) infections of a wide array of
apicomplexan parasites.
[0223] In further aspects, compounds provided herein and
derivatives thereof can be used as microbicides effective against
infections in numerous insect hosts of apicomplexan parasites
(e.g., mosquitoes, ticks).
[0224] Therapeutic intervention of apicomplexan parasites is
generally focused on the inhibition of metabolic processes.
Frequently targeted are the nucleotide salvage pathways (e.g.,
folate biosynthesis) or the apicoplast, a non-photosynthetic
remnant of a chloroplast that is sensitive to certain antibiotics
and is also involved in fatty acid and isoprenoid synthesis.
Particularly troubling is the rapid rate of drug resistance
(particularly in the malarial parasite Plasmodium falciparum), as
well as the prohibitively high costs and toxicity of newly
developed drugs. In addition, because the present compounds target
membrane fusion events, it is highly likely to be effective against
a wide array of these parasites and less likely to enable the
parasites to develop resistance.
[0225] In spite of the conserved mechanism of host cell invasion in
this phylum, current therapies do not target the invasive machinery
which is composed of a parasite actin:myosin motor linked to
organellar adhesins that are secreted at the onset of invasion.
Small molecule screens have identified inhibitors of invasion
(e.g., Carey K L et al, Proc Natl Acad Sci USA. 2004, 101:7433-8)
but their precise targets and mechanism are unknown and they have
not been utilized therapeutically in human or animal infections.
Other inhibitors of invasion have been identified (Kato et al.,
Nature Chemical Biology, 4: 347-356 (2008)), but they are clearly
distinct from the novel process identified here. These parasites do
not have an extracellular life cycle, thus disruption of the
invasive machinery is lethal.
[0226] The unique mechanism of inhibition of membrane fusion events
described herein combined with data demonstrating the inhibition of
T. gondii attachment and motility provides a novel approach for
controlling apicomplexan parasites. Taken together, these data
indicate that compounds provided herein act by inhibiting fusion of
the Toxoplasma invasive organelles at the onset of host cell entry,
arresting parasites in an extracellular environment in which they
cannot survive. Existing therapies that target metabolic pathways
or bacterial-like components of the apicoplast are prone to
developing rapid drug resistance, thus compounds that targets lipid
fusion events provide an attractive mechanism for targeting a wide
array of these parasites and also avoiding rapid evolution of
resistance.
[0227] Compounds provided herein are useful for inhibiting
infections by other apicomplexan pathogens that affect humans
(e.g., Toxoplasma, Plasmodium spp., Cryptosporidium spp., and
Isospora belli) as well as pathogens of veterinary interest (e.g.,
Theileria spp, Babesia spp., Eimeria spp., Neospora caninum).
[0228] Also provided herein are methods for inactivating parasites
in a biological or pharmaceutical preparation, the methods
comprising adding an antiparasitic compound provided herein to the
preparation and incubating the mixture for a time sufficient to
inactivate parasites present in the preparation.
[0229] In some preferred aspects, the antiparasitic compounds are
substantially inert with respect to the structure and function of
macromolecules, cells, tissues, organs and/or other biological
structures comprising the preparation. For example, in some
aspects, treating biological preparations with the antiparasitic
compounds at a concentration and time sufficient to inactivate
parasites within the preparation does not result in detectable
protein denaturation, protein degradation, plasma membrane
disruption, cell lysis, or the like.
[0230] In additional aspects, biological preparations treated with
the antiparasitic compounds, at a concentration and time sufficient
to inactivate parasites within the preparation, are substantially
non-toxic to subjects to whom the preparations are intended for
delivery, including but not limited to, human subjects. In some
preferred aspects, biological preparations treated with the
antiparasitic compounds are substantially non-toxic to human
subjects without the need for further purification or
processing.
[0231] In some aspects, the biological preparation is a biological
sample drawn from a human or animal donor, such as but not limited
to, blood, plasma, cerebrospinal fluid, mammary fluid, embryonic
fluid, mucus, urine, and the like. For example, in some aspects,
blood, tissue, or an organ harvested from a human or animal donor
is treated according to methods provided herein to inactivate
parasites prior to transplantation into a human or animal
recipient. In some preferred aspects, the treated donor sample is
transplanted into the recipient without removing the antiparasitic
compound(s).
[0232] In further aspects, the biological preparation comprises
cultured cells, tissues, or organs, such as but not limited to stem
cells or xcnographic tissues intended for transplantation. In
additional aspects, the biological preparation comprises cultured
host cells for the production of a recombinant protein or other
biological product. In some preferred aspects, treating a cellular
preparation with an antiparasitic compound provided herein
inactivates parasites within the preparation without substantially
affecting the growth, proliferation, viability, and/or productivity
of the cells. In some preferred aspects, the treated cells are used
or harvested without removing the antiparasitic compound(s).
[0233] Also provided herein are kits comprising a container housing
an antiparasitic compound provided herein and instructions for
administering the compound to a subject that has been infected or
is at risk of infection by a parasite. The instructions can provide
for administration as an oral formulation, by inhalation, by
topical administration, by intravenous injection and/or by any
other suitable means.
[0234] In some aspects, kits provided herein optionally further
comprise a pharmaceutical preparation vial and a pharmaceutically
acceptable diluent, such as physiological saline for diluting a
concentrated solution, salt or lyophilized powder formulation of an
antiparasitic compound provided herein.
[0235] In some aspects, the kit comprises an inhaler for
aerosolized administration to the lungs and/or upper respiratory
tract. In further aspects, the kit comprises a device for
intranasal administration to the nasal mucosa.
Exemplary Aspects
EXAMPLE 1
Compound Preparation
[0236] General Reaction
[0237] (Z)
3-ethyl-5-(5-phenylfuran-2-ylmethyl-ene)oxazolidine-2,4-dithion- e
1 (JL102) and (Z)
3-ethyl-5-[5-(2-methoxyphenyl)-furan-2-ylmethylene]oxazolidine-2,4-dithio-
ne 2 (JL103).
##STR00021##
[0238] The two compounds were prepared from readily available
materials as described in detail below. For compound 1 (JL102), the
readily available ethyl 2-thioxooxazolidin-4-one 3 was reacted with
the 5-phenyl-2-furfuraldehyde 4 in the presence of sodium acetate
and acetic acid to produce the N-ethyl
5-(5-phenyl-2-furanylmethylene) 2-thioxooxazolidin-4-one 5 (JL101).
Reaction of 5 with commercially available Lawesson's reagent in
refluxing toluene afforded the desire oxazolidine-2,4-dithione 1
(JL102).
##STR00022##
[0239] The second compound 2 (JL103) was prepared by similar
chemistry. First the N-ethyl oxazolidine-2,4-dithione 6 was
prepared from the 2-thioxooxazolidin-4-one 3 with Lawesson's
reagent. Also the 5-(2-methoxyphenyl)-2-furfuraldehyde 9 was
prepared by the Suzuki coupling of the bromide 7 with the boronic
acid 8. Finally reaction of the N-ethyl oxazolidine-2,4-dithione 6
with the aldehyde 9 (using sodium acetate and acetic acid) gave the
desired 5-arylmethylidene oxazolidine-2,4-dithione 2 (JL103). Many
substituted derivatives of this class of compounds could be easily
prepared by reaction of the N-substituted oxazolidine-2,4-dithiones
10 with various 5-aryl substituted 2-furfuraldehydes 11 using
sodium acetate and acetic acid (or another base/acid combination)
to give the desired N-substituted 5-arylmethylidene
oxazolidine-2,4-dithiones 12.
##STR00023##
General Procedures
[0240] Materials were obtained from commercial suppliers and were
used without further purification. Air or moisture sensitive
reactions were conducted under argon atmosphere using oven-dried
glassware and standard syringe/septa techniques. The reactions were
monitored with a silica gel TLC plate under UV light (254 nm)
followed by visualization with a p-anisaldehyde, ninhydrin, or
Cerium Molybdate (Hanessian's stain) staining solution. Column
chromatography was performed on silica gel 60. .sup.1H NMR spectra
were measured at 400 MHz in CDCl.sub.3 unless stated otherwise and
data were reported as follows in ppm (.delta.) from the internal
standard (TMS, 0.0 ppm): chemical shift (multiplicity, integration,
coupling constant in Hz.).
Preparation of (Z)
3-Ethyl-5-(5-phenylfuran-2-ylmethylene)-2-thioxooxazolidin-4-one 5
(JL101)
##STR00024##
[0242] To a mixture of 3-ethyl-2-thioxooxazolidin-4-one 3 (145 mg,
1.0 mmol), 5-phenyl-2-furaldehyde 4 (172 mg, 1.0 mmol) and sodium
acetate (410 mg, 5.0 mmol) was added acetic acid (5 mL). The
reaction mixture was refluxed for 30 h, then the excess solvent was
removed in vacuo. The residue was diluted with ethyl acetate (100
mL), then washed with water (2.times.20 mL). The organic layer was
dried with brine and MgSO.sub.4, then concentrated in vacuo. The
desired product 5 (JL101) was obtained by recrystallization from an
ethyl acetate/hexanes mixture in 45% yield (135 mg, light yellow
needles). .sup.1H NMR (400 MHz, CDCl.sub.3): 7.81-7.76 (m, 2H),
7.44 (dt, J=8.0, 1.6 Hz), 7.36 (t, J=6.0 Hz, 1H), 7.28 (d, J-3.6
Hz, 1H), 6.88 (d, J-3.6 Hz, 1H), 6.80 (s, 1H), 3.97 (q, J-7.2 Hz,
2H), 1.34 (t, J-7.2 Hz, 3H).
Preparation of (Z)
3-Ethyl-5-(5-phenylfuran-2-ylmethylene)oxazolidine-2,4-dithione 1
(JL102)
##STR00025##
[0244] A toluene (3 mL) solution of
3-ethyl-5-(5-phenylfuran-2-ylmethylene)-2-thioxooxazolidin-4-one 5
(JL101) (88 mg, 0.29 mmol) and Lawesson's reagent (176 mg, 0.435
mmol) was refluxed overnight. The solution was cooled and diluted
with ethyl acetate (50 mL), then washed with water (2.times.10 mL).
The organic layer was dried with brine and MgSO.sub.4, and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (hexane:ethyl acetate=20:1), and the
desired product 1 (JL102) was obtained in 82% yield (75 mg,
red-orange solid). .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.80
(d, J=1.2 Hz, 2H), 7.46-7.36 (m, 4H), 7.05 (s, 1H), 6.91 (d, J=3.6
Hz, 1H), 4.30 (q, J=7.2 Hz, 2H), 1.34 (t, J=7.2 Hz, 3H); .sup.13C
NMR (100 MHz, CDCl.sub.3): .delta. 182.8, 182.3, 158.5, 148.5,
147.0, 129.3, 129.1, 124.8, 122.3, 117.6, 109.9, 103.0, 41.7, 11.3
ppm; IR (NaCl) 2917, 2848, 1634, 1389, 1360, 1348, 1271, 1226,
1114, 1078, 1033, 953, 760, 691 cm.sup.-1.
Preparation of 3-Ethyloxazolidine-2,4-dithione (6)
##STR00026##
[0246] A toluene (10 mL) solution of
3-ethyl-2-thioxooxazolidin-4-one 3 (145 mg, 1.0 mmol) and
Lawesson's reagent (607 mg, 1.5 mmol) was refluxed overnight. The
solution was cooled and diluted with ethyl acetate (100 mL), then
washed with water (2.times.20 mL). The organic layer was dried with
brine and MgSO.sub.4, and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel (hexane:ethyl
acetate=10:1), and the desired product 6 was obtained in 94% yield
(152 mg, orange oil). .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
5.12 (s, 2H), 4.21 (q, J=7.2 Hz, 2H), 1.29 (t, J=7.2 Hz, 3H).
Preparation of Aryl-2-furaldehydes
[0247] Aryl-2-furaldehydes were generally prepared accordingly:
##STR00027##
[0248] To a benzene (10 mL) solution of 5-bromo-2-furaldehyde 7
(350 mg, 2.0 mmol) and arylboronic acid 27 (2.0 mmol) was added
tetrakis(triphenylphosphine)-palladium(0) (69.4 mg, 0.06 mmol) and
sat. aq. K.sub.2CO.sub.3 solution (2.0 ml) at 21.degree. C. under
an argon atmosphere. The reaction mixture was refluxed for
overnight. The mixture was then cooled and diluted with ethyl
acetate (100 mL). The organic layer was washed with water
(2.times.20 mL), then dried with brine and MgSO.sub.4 and
concentrated in vacuo. Flash column chromatography on silica gel
(n-hexanes:ethyl acetate-20:1) of the residue afforded
5-aryl-2-furaldehyde.
[0249] The following compounds were prepared in accordance with the
above scheme.
TABLE-US-00001 R' Yield (%) 9 2-methoxy 65 28 4-methoxy 78 29
2,4-dimethoxy 58 30 2,3-dimethoxy 80 31 2,6-dimethoxy 26 32
3-methoxy 87 33 4-dimethylamino 66 34 2,4,6-trimethoxy 32
Preparation of 5-(2-Methoxyphenyl)-2-furaldehyde (9)
##STR00028##
[0251] To a benzene (10 mL) solution of 5-bromo-2-furaldehyde 7
(350 mg, 2.0 mmol) and 2-methoxyphenylboronic acid 8 (304 mg, 2.0
mmol) was added tetrakis(triphenylphosphine)-palladium(0) (69.4 mg,
0.06 mmol) and sat. aq. K.sub.2CO.sub.3 solution (4.94 ml, 4.0
mmol) at 21.degree. C. under an argon atmosphere. The reaction
mixture was refluxed for 2 h. The mixture was then cooled and
diluted with ethyl acetate (100 mL). The organic layer was washed
with water (2.times.20 mL), then dried with brine and MgSO.sub.4
and concentrated in vacuo. Flash column chromatography on silica
gel (hexane:ethyl acetate=20:1) of the residue afforded
5-(2-methoxyphenyl)-2-furaldehyde 9 in 65% yield (265 mg, yellow
oil). .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.65 (s, 1H), 8.05
(dd, J=8.0, 2.0 Hz, 1H), 7.39-7.35 (m, 1H), 7.33 (d, J=3.6 Hz, 1H),
7.14 (d, J=4.0 Hz, 1H), 7.07 (dt, J=7.2, 0.8 Hz, 1H), 7.00 (bd,
J=8.0 Hz, 1H), 3.97 (s, 3H).
[0252] Characterization data for compounds 28-34 are discussed
below.
[0253] 5-(4-Methoxyphenyl)-2-furaldehyde 28: yellow caramel.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.60 (s, 1H), 7.77 (d,
J=8.8 Hz, 1H), 7.31 (d, J=3.6 Hz, 1H), 6.97 (d, J=8.8 Hz, 1H), 6.72
(d, J=3.6 Hz, 1H), 3.86 (s, 3H).
[0254] 5-(2,4-Dimethoxyphenyl)-2-furaldehyde 29: yellow caramel.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.59 (s, 1H), 7.98 (d,
J=8.8 Hz, 1H), 7.31 (d, J=3.6 Hz, 1H), 7.00 (d, J=3.6 Hz, 1H), 6.60
(dd, J=8.8, 2.4 Hz, 1H), 6.54 (d, J=2.4 Hz, 1H), 3.94 (s, 3H), 3.87
(s, 3H).
[0255] 5-(2,3-Dimethoxyphenyl)-2-furaldehyde 30: light yellow
caramel. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.66 (s, 1H),
7.59 (dd, J=8.0, 1.6 Hz, 1H), 7.34 (d, J=3.6 Hz, 1H), 7.18 (d,
J=3.6 Hz, 1H), 7.15 (t, J=8.0 Hz, 1H), 6.97 (dd, J=8.0, 1.6 Hz,
1H), 3.91 (s, 3H), 3.89 (s, 3H).
[0256] 5-(2,6-Dimethoxyphenyl)-2-furaldehyde 31: yellow caramel.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.66 (s, 1H), 7.35 (t,
J=8.4 Hz, 1H), 7.34 (d, J=3.6 Hz, 1H), 6.74 (d, J=3.6 Hz, 1H), 6.63
(d, J=8.4 Hz, 2H), 3.83 (s, 6H).
[0257] 5-(3-Methoxyphenyl)-2-furaldehyde 32: yellow oil. .sup.1H
NMR (400 MHz, CDCl.sub.3): .delta. 9.66 (s, 1H), 7.42-7.35 (m, 3H),
7.32 (d, J=3.6 Hz, 1H), 6.95 (ddd, J=8.0, 3.2, 0.8 Hz, 1H), 6.84
(d, J=3.6 Hz, 1H), 3.88 (s, 3H).
[0258] 5-(4-Dimethoxyphenyl)-2-furaldehyde 33: light yellow solid.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.54 (s, 1H), 7.71 (d,
J=8.8 Hz, 2H), 7.29 (d, J=3.6 Hz, 1H), 6.73 (d, J=8.8 Hz, 2H), 6.63
(d, J=3.6 Hz, 1H), 3.04 (s, 6H).
[0259] 5-(2,4,6-Trimethoxyphenyl)-2-furaldehyde 34: yellow solid.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.62 (s, 1H), 7.32 (d,
J=3.6 Hz, 1H), 6.68 (d, J=3.6 Hz, 1H), 6.18 (S, 2H), 3.86 (s, 3H),
3.82 (s, 6H).
Preparation of
(Z)-3-Ethyl-5-(5-aryl-furan-2-ylmethylene)oxazolidine-2,4-dithiones
[0260]
(Z)-3-Ethyl-5-(5-aryl-furan-2-ylmethylene)oxazolidine-2,4-dithiones
were generally prepared accordingly:
##STR00029##
[0261] To a mixture of 3-ethyloxazolidine-2,4-dithione 6 (79 mg,
0.49 mmol), 5-(2-methoxyphenyl)-2-furaldehyde 9 (99 mg, 0.49 mmol)
and sodium acetate (201 mg, 2.45 mmol) was added acetic acid (5
mL). As more of the desired product was formed, the reaction
mixture color turned to a very dark red solution. The reaction
mixture was refluxed overnight, then the excess solvent was removed
in vacuo. The residue was diluted with ethyl acetate (100 mL), then
washed with water (2.times.20 mL). The organic layer was dried with
brine and MgSO.sub.4, then concentrated in vacua. The desired
product was prepared by recrystallization from ethyl
acetate/n-hexanes.
[0262] The following compounds were prepared in accordance with the
above scheme.
TABLE-US-00002 R' Yield (%) 1 JL102 H 87 2 JL103 2-methoxy 95 16
JL108 4-methoxy 80 17 JL109 2,4-dimethoxy 93 18 JL110 2,3-dimethoxy
83 19 JL111 2,6-dimethoxy 90 22 JL117 3-methoxy 73 23 JL118
4-dimethylamino 70 26 JL122 2,4,6-trimethoxy 69
Preparation of (Z)
3-Ethyl-5-[5-(2-methoxyphenyl)-furan-2-ylmethylene]oxazolidine-2,4-dithio-
ne 2 (JL103)
##STR00030##
[0264] To a mixture of 3-ethyloxazolidine-2,4-dithione 6 (79 mg,
0.49 mmol), 5-(2-methoxyphenyI)-2-furaldehyde 9 (99 mg, 0.49 mmol)
and sodium acetate (201 mg, 2.45 mmol) was added acetic acid (5
mL). As more of the desired product was formed, the reaction
mixture color turned to a very red solution. The reaction mixture
was refluxed overnight, then the excess solvent was removed in
vacua. The residue was diluted with ethyl acetate (100 mL), then
washed with water (2.times.20 mL). The organic layer was dried with
brine and MgSO.sub.4, then concentrated in vacua. Flash column
chromatography afforded the desired product 2 (JL103) in 95% yield
(160 mg, purple solid).
[0265] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.98 (dd, J=8.0,
1.6 Hz, 1H), 7.37-7.33 (m, 2H), 7.19 (d, J=4.0 Hz, 1H), 7.08 (t,
J=3.6 Hz, 1H), 7.05 (s, 1H), 6.98 (bd, J=8.4 Hz, 1H), 4.30 (q,
J=7.2 Hz, 2H), 3.97 (s, 3H), 1.34 (t, J=7.2 Hz, 3H); .sup.13C NMR
(100 MHz, CDCl.sub.3): .delta. 182.6, 182.3, 156.5, 155.5, 147.5,
146.8, 130.2, 127.0, 122.9, 121.1, 118.3, 114.9, 111.2, 103.3,
55.5, 41.6, 11.3 ppm; IR (NaCl) 2967, 2921, 2845, 1622, 1505, 1486,
1391, 1351, 1304, 1271, 1245, 1114, 1078, 1022, 952, 788, 752
cm.sup.-1; MS (EI.sup.+): m/z 346 [M+H].sup.+; HRMS calculated for
C.sub.17H.sub.15NO.sub.3S.sub.2: 345.0493; Found: 346.0544
[M+H].sup.+.
[0266] Characterization data for compounds 1, 16-19, 22, 23 and 26
are discussed below.
[0267]
(Z)-3-Ethyl-5-(5-phenylfuran-2-ylmethylene)oxazolidine-2,4-dithione
1 (JL102): red-orange solid. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 7.80 (d, J=1.2 Hz, 2H), 7.46-7.36 (m, 4H), 7.05 (s, 1H),
6.91 (d, J=3.6 Hz, 1H), 4.30 (q, J=7.2 Hz, 2H), 1.34 (t, J=7.2 Hz,
3H); .sup.13C NMR (100 MHz, CDCl.sub.3): .delta. 182.8, 182.3,
158.5, 148.5, 147.0, 129.3, 129.1, 124.8, 122.3, 117.6, 109.9,
103.0, 41.7, 11.3 ppm; IR (NaCl) 2917, 2848, 1634, 1389, 1360,
1348, 1271, 1226, 1114, 1078, 1033, 953, 760, 691 cm.sup.-1.
[0268]
(Z)-3-Ethyl-5-[5-(2-methoxyphenyl)-furan-2-ylmethylene]oxazolidine--
2,4-dithione 2 (JL103): purple solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 7.98 (dd, J=8.0, 1.6 Hz, 1H), 7.37-7.33 (m,
2H), 7.19 (d, J=4.0 Hz, 1H), 7.08 (t, J=3.6 Hz, 1H), 7.05 (s, 1H),
6.98 (bd, J-8.4 Hz, 1H), 4.30 (q, J-7.2 Hz, 2H), 3.97 (s, 3H), 1.34
(t, J-7.2 Hz, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3): .delta.
182.6, 182.3, 156.5, 155.5, 147.5, 146.8, 130.2, 127.0, 122.9,
121.1, 118.3, 114.9, 111.2, 103.3, 55.5, 41.6, 11.3 ppm; IR (NaCl)
2967, 2921, 2845, 1622, 1505, 1486, 1391, 1351, 1304, 1271, 1245,
1114, 1078, 1022, 952, 788, 752 cm.sup.-1; MS (EI.sup.-1): m/z 346
[M+H].sup.+; HRMS calculated for C.sub.17H.sub.15NO.sub.3S.sub.2:
345.0493; Found: 346.0544 [M+H].sup.+.
[0269]
(Z)-3-Ethyl-5-(5-(4-methoxyphenyl)-furan-2-ylmethylene)oxazolidine--
2,4-dithione 16 (JL108): purple solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 7.74 (d, J=9.2 Hz, 2H), 7.36 (d, J=3.6 Hz,
1H), 7.06 (s, 1H), 6.98 (d, J=9.2 Hz, 2H), 6.80 (d, J=3.6 Hz, 1H),
4.30 (q, J=7.2 Hz, 2H), 3.87 (s, 3H), 1.35 (t, J=7.2 Hz, 3H).
[0270]
(Z)-3-Ethyl-5-(5-(2,4-dimethoxyphenyl)-furan-2-ylmethylene)oxazolid-
ine-2,4-dithione 17 (JL109): dark purple solid. .sup.1H NMR (400
MHz, CDCl.sub.3): .delta. 7.97 (d, J=4.0 Hz, 1H), 7.36 (d, J=3.6
Hz, 1H), 7.08 (d, J=3.6 Hz, 1H), 7.06 (s, 1H), 6.64 (dd, J=8.8, 2.4
Hz, 1H), 6.53 (d, J=2.4 Hz, 1H), 4.30 (q, J=7.2 Hz, 2H), 4.05 (s,
3H), 3.95 (s, 3H), 1.34 (t, J=7.2 Hz, 3H).
[0271]
(Z)-3-Ethyl-5-(5-(2,3-dimethoxyphenyl)-furan-2-ylmethylene)oxazolid-
ine-2,4-dithione 18 (JL110): dark purple solid. .sup.1H NMR (400
MHz, CDCl.sub.3): .delta. 7.55 (dd, J=8.0, 1.6 Hz, 1H), 7.38 (d,
J-3.6 Hz, 1H), 7.23 (d, J-3.6 Hz, 1H), 7.17 (t, J-8.0 Hz, 1H), 7.05
(s, 1H), 6.95 (dd, J=8.0, 1.6 Hz, 1H), 4.31 (q, J=7.2 Hz, 2H), 3.92
(s, 3H), 3.90 (s, 3H), 1.35 (t, J=7.2 Hz, 3H).
[0272]
(Z)-3-Ethyl-5-(5-(2,6-dimethoxyphenyl)-furan-2-ylmethylene)oxazolid-
ine-2,4-dithione 19 (JL111): dark purple solid. .sup.1H NMR (400
MHz, CDCl.sub.3): .delta. 7.38 (d, J=3.6 Hz, 1H), 7.33 (t, J=8.4
Hz, 1H), 7.13 (s, 1H), 6.95 (d, J=3.6 Hz, 1H), 6.65 (d, J=8.4 Hz,
2H), 4.30 (q, J=7.2 Hz, 2H), 4.05 (s, 3H), 3.91 (s, 6H), 1.34 (t,
J=7.2 Hz, 3H).
[0273]
(Z)-3-Ethyl-5-(5-(3-methoxyphenyl)-furan-2-ylmethylene)oxazolidine--
2,4-dithione 22 (JL117): dark red solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 7.39-7.33 (m, 4H), 7.05 (s, 1H), 6.95-6.92 (m,
1H), 6.91 (d, J=3.6 Hz, 1H), 4.31 (q, J=7.2 Hz, 2H), 3.90 (s, 3H),
1.35 (t, J=7.2 Hz, 3H).
[0274]
(Z)-3-Ethyl-5-(5-(4-dimethylaminophenyl)-furan-2-ylmethylene)oxazol-
idine-2,4-dithione 23 (JL118): dark purple solid. .sup.1H NMR (400
MHz, CDCl.sub.3): .delta. 7.68 (d, J=8.8 Hz, 2H), 7.38 (d, J=3.6
Hz, 1H), 7.07 (s, 1H), 6.76-6.73 (m, 3H), 4.30 (q, J=7.2 Hz, 2H),
3.05 (s, 6H), 1.35 (t, J=7.2 Hz, 3H).
[0275]
(Z)-3-Ethyl-5-(5-(2,4,6-tiimethoxyphenyl)-furan-2-ylmethylene)oxazo-
lidine-2,4-dithione 26 (JL122): black solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 7.36 (d, J=3.6 Hz, 1H), 7.08 (s, 1H), 6.90 (d,
J=3.6 Hz, 1H), 6.21 (s, 2H), 4.30 (q, J=7.2 Hz, 2H), 3.91 (s, 6H),
3.87 (s, 3H), 1.34 (t, J=7.2 Hz, 3H).
Preparation of JL104
[0276] JL104 was prepared from readily available materials
according to the following scheme, as described in detail below.
Similar compounds may also be prepared using this scheme.
##STR00031##
Preparation of 3-ethyl-2-thioxoimidazolidin-4-one 37
[0277] Glycinc 36 (150 mg, 2.0 mmol) was dissolved in H.sub.2O (5.0
mL) and pyridine (5.0 mL) solution. The pH of the solution was
adjusted to about 9.0 as shown by an indicator paper by the
addition of 1M NaOH. The solution was heated to 40.degree. C. and
kept at that temperature during the reaction. Ethyl isothiocyanate
35 (0.263 mL, 3.0 mmol) was added with vigorous stirring. Small
portions of 1M NaOH was added to keep the pH at about 9.0. The
reaction was completed when the alkali consumption ceased (1 h).
Pyridine and excess alkyl isothiocyanates were then removed by
repeated extraction with equal volumes of benzene. Subsequently, an
amount of conc. HCl (0.75 mL) was added and the mixture was
refluxed for 2 h. The residue was extracted with ethyl acetate
(2.times.150 mL). The organic layer was dried with brine and
MgSO.sub.4, then concentrated in vacuo. Flash column chromatography
on silica gel (hexane:ethyl acetate=2:1) of the residue afforded
the desired product 37 in 90% yield (258 mg, pale yellow solid).
.sup.1H NMR (400 MHz, CDCl.sub.3): 6.88 (br, 1H), 4.06 (d, J=1.2
Hz, 2H), 3.89 (q, J=7.2 Hz, 2H), 1.25 (t, J=7.2 Hz, 3H).
Preparation of
(Z)-3-ethyl-5-((5-phenylfuran-2-yl)methylene)-2-thioxoimidazolidin-4-one
13 (JL104)
[0278] To an ethanol (10 mL) solution of
3-ethyl-2-thioxoimidazolidin-4-one 37 (145 mg, 1.0 mmol),
5-phenyl-2-furaldehyde 4 (172 mg, 1.0 mmol) was added pyridine
(0.243 mL, 3.0 mmol). The reaction mixture was refluxed overnight,
then the excess solvent was removed in vacuo. The residue was
diluted with ethyl acetate (100 mL), then washed with water
(2.times.20 mL). The organic layer was dried with brine and
MgSO.sub.4, then concentrated in vacuo. The desired product 13 (JL
104) was obtained by recrystallization from an ethyl
acetate/hexanes mixture in 50% yield (149 mg, yellow solid).
.sup.1H NMR (400 MHz, CDCl.sub.3): 9.16 (br, 1H), 7.69 (dd, J=7.2,
1.6 Hz, 2H), 7.49 (t, J=7.2 Hz, 2H), 7.41 (t, J=7.2 Hz, 1H), 6.52
(s, 1H), 4.00 (q, J=7.2 Hz, 2H), 1.31 (t, J=7.2 Hz, 3H).
Preparation of JL105
[0279] JL105 was prepared from readily available materials
according to the following scheme, as described in detail below.
Similar compounds may also be prepared using this scheme.
##STR00032##
Preparation of
(Z)-3-ethyl-5-((5-phenylfuran-2-yl)methylene)imidazolidine-2,4-dithione
14 (JL105)
[0280] A toluene (1 mL) solution of
(Z)-3-ethyl-5-((5-phenylfuran-2-yl)methylene)-2-thioxoimidazolidin-4-one
13 (JL104) (30 mg, 0.1 mmol) and Lawesson's reagent (61 mg, 0.15
mmol) was refluxed overnight. The solution was cooled and diluted
with ethyl acetate (50 mL), then washed with water (2.times.10 mL).
The organic layer was dried with brine and MgSO.sub.4, and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (n-hexanes:ethyl acetate=20:1), and
the desired product 14 (JL105) was obtained in 89% yield (28 mg,
red-orange solid). .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.47
(br, 1H), 7.71 (d, J=7.2 Hz, 2H), 7.49 (t, J=7.2 Hz, 2H), 7.40 (t,
J=7.2 Hz, 1H), 6.95 (d, J=3.6 Hz, 1H), 6.85 (d, J=3.6 Hz, 1H), 6.84
(s, 1H), 4.36 (q, J=7.2 Hz, 2H), 1.34 (t, J=7.2 Hz, 3H
Preparation of JL107
[0281] JL107 was prepared from readily available materials
according to the following scheme, as described in detail below.
Similar compounds may also be prepared using this scheme.
##STR00033##
[0282] To a DMF (5 mL) solution of sodium borohydride (11.7 mg,
0.31 mmol) was added
(Z)-3-Ethyl-5-(5-phenylfuran-2-ylmethylene)oxazolidine-2,4-dith-
ione 1 (130mg, 0.41 mmol) in DMF (2 mL) dropwise at 0.degree. C.
under argon atmosphere. As the reaction proceeded, the reaction
misture color was lightend to yellowish solution. The mixture was
stirred for 1 h at 0.degree. C. The reaction solution was poured
into 1% aq. HCl, then extracted with ethyl acetate (2.times.50 mL).
The organic layer was dried with brine and MgSO.sub.4, and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (n-hexanes:ethyl acetate=30:1), and
the desired product 15 (JL107) was obtained in 54% yield (70 mg,
orange caramel). .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.61
(d, J=7.2 Hz, 2H), 7.37 (t, J=7.2 Hz, 2H), 7.28-7.23 (m, 1H), 6.56
(d, J=3.2 Hz, 1H), 6.25 (d, J=3.2 Hz, 1H), 3.29 (dd, J=12.8, 6.8
Hz, 1H), 4.77 (dd, J=9.2, 3.6 Hz, 1H), 4.06 (qd, J=7.2, 3.6 Hz,
2H), 3.91 (dd, J=14.8, 3.2 Hz, 1H), 1.18 (t, J=7.2 Hz, 3H).
Preparation of JL113
[0283] JL113 was prepared from readily available materials
according to the following scheme, as described in detail below.
Similar compounds may also be prepared using this scheme.
##STR00034##
Preparation of 2,3-Benzofuran-5-carboxaldehyde 39
[0284] To a DMF (0.580 mL) solution of 2,3-benzofuran 38 (0.331 mL,
3.0 mmol) was added phosphorous oxychloride (0.302 mL, 3.3 mmol).
The reaction mixture was stirred overnight at 80.degree. C., then
the excess solvent was removed in vacuo. The residue was diluted
with ethyl acetate (100 mL), then washed with water (2.times.20
mL). The organic layer was dried with brine and MgSO.sub.4, then
concentrated in vacuo. Flash column chromatography on silica gel
(n-hexanes:ethyl acetate=50:1) of the residue afforded the desired
product 39 in 40% yield (172 mg, yellow oil). .sup.1H NMR (400 MHz,
CDCl.sub.3): 9.88 (s, 1H), 7.76 (d, J=7.2 Hz, 2H), 7.62 (dd, J=8.4,
0.8 Hz, 1H), 7.54 (d, J=0.8 Hz, 1H), 7.53 (td, J=7.2, 1.2 Hz, 1H),
7.35 (td, J=7.2, 0.8 Hz, 1H).
Preparation of
(Z)-5-(benzofuran-2-ylmethylene)-3-ethyloxazolidine-2,4-dithione 20
(JL113)
[0285] To a mixture of 3-ethyloxazolidine-2,4-dithione 6 (100 mg,
0.62 mmol), 2,3-benzofuran-5-carboxaldehyde 39 (91 mg, 0.62 mmol)
and sodium acetate (254 mg, 3.10 mmol) was added acetic acid (6
mL). The reaction mixture was refluxed for 1 h, then the excess
solvent was removed in vacuo. The residue was diluted with ethyl
acetate (70 mL), then washed with water (2.times.20 mL). The
organic layer was dried with brine and MgSO.sub.4, then
concentrated in vacuo. Flash column chromatography on silica gel
(n-hexanes:ethyl acetate=10:1) of the residue afforded the desired
product 20 (JL113) in 80% yield (144 mg, red-orange solid). .sup.1H
NMR (400 MHz, CDCl.sub.3): 7.68 (d, J=3.6 Hz, 1H), 7.57 (s, 1H),
7.54 (dd, J=8.4, 0.8 Hz, 1H), 7.43 (td, J=7.2, 1.2 Hz, 1H), 7.29
(td, J=7.2, 1.2 Hz, 1H), 7.07 (s, 1H), 4.32 (q, J=7.2 Hz, 2H), 1.36
(t, J=7.2 Hz, 3H).
Preparation of JL119 and JL123 and Similar Compounds
[0286] JL119 and JL123 were prepared from readily available
materials according to the following scheme, as described in detail
below. Similar compounds may also be prepared using this
scheme.
##STR00035##
Preparation of 1-Methyl-5-aryl-1H-pyrrole-2-carboxaldehydes
[0287] 1-Methyl-5-aryl-1H-pyrrole-2-carboxaldehydes were generally
prepared accordingly:
##STR00036##
[0288] To a mixture of 1-methyl-2-pyrrole-carboxaldehyde 40 (0.408
mL. 4.0 mmol), substituted arylbromide (2.0 mmol), palladium
diacetate (2.25 mg, 0.01 mmol) and K.sub.2CO.sub.3 (552 mg, 4.0
mmol) was added DMAc (6 mL) at 21.degree. C. under an argon
atmosphere. The reaction mixture was refluxed for overnight. The
mixture was then cooled and diluted with ethyl acetate (100 mL).
The organic layer was washed with water (2.times.20 mL), then dried
with brine and MgSO.sub.4 and concentrated in vacuo. Flash column
chromatography on silica gel (n-hexanes:ethyl acetate=20:1) of the
residue afforded 1-methyl-5-aryl-1H-pyrrole-2-carboxaldehyde.
[0289] The following compounds were prepared in accordance with the
above scheme.
TABLE-US-00003 R' Yield (%) 44 4-methoxy 60 45 4-dimethylamino
20
[0290] Characterization data for compounds 44 and 45 are discussed
below.
[0291] 5-(4-Methoxyphenyl)-1-methyl-1H-pyrrole-2-carboxaldehyde 44:
60% yield, pale yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 9.55 (s, 1H), 7.35 (d, J=8.8 Hz, 2H), 6.99 (d, J=8.8 Hz,
2H), 6.97 (d, J=4.0 Hz, 1H), 6.26 (d, J=4.0 Hz, 1H), 3.92 (s, 3H),
3.87 (s, 3H).
[0292]
5-(4-Dimethylaminophenyl)-1-methyl-1H-pyrrole-2-carboxaldehyde 45:
20% yield, brown solid. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
9.51 (s, 1H), 7.30 (d, J=8.8 Hz, 2H), 6.95 (d, J=4.0 Hz, 1H), 6.77
(d, J=8.8 Hz, 2H), 6.24 (d, J=4.0 Hz, 1H), 3.94 (s, 3H), 3.02 (s,
6H).
Preparation of
(Z)-3-Ethyl-5-((1-methyl-5-aryl-1H-pyrrole-2-yl)methylene)oxazolidine-2,4-
-dithiones
[0293]
(Z)-3-Ethyl-5-((1-methyl-5-aryl-1H-pyrrole-2-yl)methylene)oxazolidi-
ne-2,4-dithiones were generally prepared accordingly:
##STR00037##
[0294] To a mixture of 3-ethyloxazolidine-2,4-dithione 6 (80 mg,
0.50 mmol), 1-Methyl-5-aryl-1H-pyrrole-2-carbox-aldehyde 42 (0.50
mmol) and sodium acetate (205 mg, 2.50 mmol) was added acetic acid
(5 mL). As more of the desired product was formed, the reaction
mixture color turned to a very dark solution. The reaction mixture
was refluxed overnight, then the excess solvent was removed in
vacuo. The residue was diluted with ethyl acetate (100 mL), then
washed with water (2.times.20 mL). The organic layer was dried with
brine and MgSO.sub.4, then concentrated in vacuo. The desired
product 43 was prepared by recrystallization from ethyl
acetate/n-hexanes.
[0295] The following compounds were prepared in accordance with the
above scheme.
TABLE-US-00004 R' Yield (%) 23 JL119 4-methoxy 89 26 JL123
4-dimethylamino 48
[0296] Characterization data for compounds 23 (JL119) and 45
(JL123) are discussed below.
[0297]
(Z)-3-Ethyl-5-((5-(4-methoxyphenyl)-1-methyl-1H-pyrrole-2-yl)methyl-
ene)oxazolidine-2,4-dithione 23 (JL119): dark purple solid. .sup.1H
NMR (400 MHz, CDCl.sub.3): .delta. 7.46 (d, J=4.0 Hz, 1H), 7.36 (d,
J=8.4 Hz, 2H), 7.11 (s, 1H), 7.00 (d, J=8.4 Hz, 2H), 6.44 (d, J=4.0
Hz, 1H), 4.31 (q,J=7.2 Hz, 2H), 3.87 (s, 3H), 3.76 (s, 3H), 1.36
(t, J=7.2 Hz, 3H).
[0298]
(Z)-3-Ethyl-5-((5-(4-methoxyphenyl)-1-methyl-1H-pyrrole-2-yl)methyl-
ene)oxazolidine-2,4-dithione 26 (JL123): black powder. .sup.1H NMR
(400 MHz, CDCl.sub.3): .delta. 7.49 (d, J=4.4 Hz, 1H), 7.32
(d,J=8.8 Hz, 2H), 7.13 (s, 1H), 6.78 (d, J=8.8 Hz, 2H), 6.45 (d,
J=4.4 Hz, 1H), 4.31 (q, J=7.2 Hz, 2H), 3.78 (s, 3H), 3.04 (s, 6H),
1.36 (t, J=7.2 Hz, 3H).
Preparation of JL121
[0299] JL121 was prepared from readily available materials
according to the following scheme, as described in detail below.
Similar compounds may also be prepared using this scheme.
##STR00038##
Preparation
(E)-2-methyl-4-((5-phenylfuran-2-yl)methylene)oxazol-5(4H)-one 47
(JL120)
[0300] To a mixture of 5-phenyl-2-furaldehyde 4 (517 mg, 3.0 mmol),
N-acetylglycine 46 (457 mg, 3.9 mmol) and sodium acetate (1230 mg,
15.0 mmol) was added acetic anhydride (30 mL). The reaction mixture
was refluxed overnight, then the excess solvent was removed in
vacuo. The residue was diluted with ethyl acetate (200 mL), then
washed with water (2.times.50 mL). The organic layer was dried with
brine and MgSO.sub.4, then concentrated in vacuo. Flash column
chromatography on silica gel (n-hexanes:ethyl acetate=10:1) of the
residue afforded the desired product 47 (JL120) in 53% yield (400
mg, orange solid). .sup.1H NMR (400 MHz, CDCl.sub.3): 7.77 (d,
J=7.2 Hz, 2H), 7.52 (d, J=3.6 Hz, 1H), 7.44 (t, J=7.2 Hz, 2H), 7.35
(t, J=7.2 Hz, 1H), 7.12 (s, 1H), 6.89 (d, J=3.6 Hz, 1H), 2.41 (s,
3H).
Preparation (Z)-Ethyl 2-hydroxy-3-(5-phenylfuran-2-yl)acrylate
48
[0301] To an ethanol (10 mL) suspension of
(E)-2-methyl-4-((5-phenylfuran-2-yl)methylene)oxazol-5(4H)-one 47
(253 mg, 1.0 mmol) was added 12M HCl (5 mL). The reaction mixture
was refluxed for 5 h, then the excess solvent was removed in vacuo.
The residue was diluted with ethyl acetate (100 mL), then washed
with water (2.times.30 mL). The organic layer was dried with brine
and MgSO.sub.4, then concentrated in vacuo. Flash column
chromatography on silica gel (n-hexanes:ethyl acetate=10:1) of the
residue afforded the desired product 48 in 55% yield (142 mg,
yellow solid). .sup.1H NMR (400 MHz, CDCl.sub.3): 7.70 (d, J=7.2
Hz, 2H), 7.39 (t, J=7.2 Hz, 2H), 7.27 (t, J=7.2 Hz, 1H), 6.97 (d,
J=3.6 Hz, 1H), 6.78 (d, J=3.6 Hz, 1H), 6.65 (d, J=1.6 Hz, 1H), 6.43
(d, J=1.6 Hz, 1H), 4.37 (q, J=7.2 Hz, 2H), 1.39 (t, J=7.2 Hz,
3H).
Preparation
(Z)-3-ethyl-5-((5-phenylfuran-2-yl)methylene)oxazolidine-2,4-dione
24 (JL121)
[0302] To an MeCN (9 mL) solution of (Z)-Ethyl
2-hydroxy-3-(5-phenylfuran-2-yl)acrylate 48 (223 mg, 0.86 mmol) was
added ethyl isocyanate 49 (0.136 mL, 1.72 mmol) and triethylamine
(0.240 mL, 1.72 mmol). The reaction mixture was stirred for 24 h,
then the excess solvent was removed in vacuo. The residue was
diluted with ethyl acetate (100 mL), then washed with water
(2.times.30 mL). The organic layer was dried with brine and
MgSO.sub.4, then concentrated in vacuo. Flash column chromatography
on silica gel (n-hexanes:ethyl acetate=10:1) of the residue
afforded the desired product 24 in 70% yield (310 mg, yellow
solid). .sup.1H NMR (400 MHz, CDCl.sub.3): 7.75 (d, J=7.2 Hz, 2H),
7.43 (t, J=7.2 Hz, 2H), 7.34 (t, J=7.2 Hz, 1H), 7.11 (d, J=3.6 Hz,
1H), 6.84 (d, J=3.6 Hz, 1H), 6.81 (s, 1H), 3.72 (q, J=7.2 Hz, 2H),
1.33 (t, J=7.2 Hz, 3H)); .sup.13C NMR (100 MHz, CDCl.sub.3):
.delta. 161.7, 160.0, 146.5, 128.9, 128.7, 124.5, 119.7, 118.0,
117.1, 116.9, 108.6, 101.9, 35.4, 13.1 ppm.
EXAMPLE 2
Biological Methods
Characterization of the New Compounds
[0303] While conducting a high-throughput screen for small molecule
inhibitors of Nipah virus (NiV) entry utilizing a pseudotyped
vesicular stomatitis virus (VSV) system, a compound was discovered
that not only inhibited VSV.DELTA.G::rhtc+NiV-F/G pseudotype
infection but also all subsequently tested enveloped virus, thus
independently of the envelope protein used by the virus to enter
the cells, but not non-enveloped viruses. This compound, a
rhodanine derivative named LJ001, inhibits viral infection with an
IC.sub.50 of .about.1 .mu.M and an IC95 of .about.10 .mu.M common
for all viruses tested [1]. A wide range of experiments allowed
demonstrating that LJ001 (1) doesn't display overt toxicity in
vitro or in vivo at concentrations of interest; (2) can prevent
virus-induced mortality in vivo, when the viruses are pretreated
with LJ001 before challenge in an animal; (3) LJ001 specifically
acts on the viral membrane and prevent infection at a step after
attachment but before virus-cell membranes fusion.
[0304] Structure-Activity relationship allowed the synthesis of new
compounds with improved antiviral activity. Compound 1's EC.sub.50
is about 10 fold lower than LJ001's, and compound 2's EC50 about 10
fold lower than compound 1's, as evaluated in an NDV infection
assay of Vero cells (FIG. 1). Without being bound by any particular
theory, it is presumed that the improved antiviral activity is
attributed to better absorption of ambient light--the absorption
peak of compound 2 is at about 500 nm whereas LJ001's is at about
440 nm, ambient light emitting most of its energy around 500 nm.
Further, compound 2 absorbs more light (see Table 1, integrated
area under the curve between 400 and 700 nm) than LJ001
(specifically, 1.53 times more). Finally, compound 2 has a higher
quantum yield--i.e., the efficiency with which one photon absorbed
from light produces its effect, (namely the generation of a singlet
oxygen)--than LJ001 (see Table 1). Of note, the correlation of
better absorption of light at longer wavelengths (see FIG. 5) and
increased singlet oxygen quantum yield to increased antiviral
activity (i.e., lower IC.sub.50 values) (Table 1), is generally
observed with respect to the compounds disclosed herein.
[0305] It is noted that the disclosed compounds are able to absorb
light at a variety of wavelengths, with some compounds approaching
near-infrared (NIR) (see Table 1, "Abs Max" and FIG. 5).
Accordingly SAR of the disclosed compounds may uncover additional
compounds which absorb light at high or even NIR wavelengths.
Cytotoxicity in the Presence of Light
[0306] Cytotoxicity evaluated by the measurement of the number of
remaining living cells (MTT) after treatment with the compound
showed that when treated in the dark the compounds are not
cytotoxic at concentrations below 10 .mu.M (FIG. 2). However when
exposed to light for different lengths of time the EC.sub.50
decreases. As an example, when exposed for 1 hour to light the
compound 1 triggers the death of 50% of the treated cells at a
concentration of .about.0.4 .mu.M. Thus, the disclosed compounds
exhibit high cytotoxicity at low concentrations upon light
exposure, with toxicity increasing with the time of exposure.
EXAMPLE 3
Antiviral Activity
[0307] The two examples of the oxazolidine-2,4-dithione class,
JL102 and JL103 (1 and 2) have strong antiviral activity at
nanomolar (nM) concentrations. These compounds are virucidal to
lipid-enveloped viruses, i.e., those that have a lipid-membrane. In
particular, we have shown that the compounds 1 and 2 inhibit the
infection of various unrelated enveloped viruses, including
Newcastle disease virus (NDV, paramyxovirus, as described in detail
below and FIG. 1), human immunodeficiency virus (HIV, retrovirus),
herpes simplex virus 1 (HSV-1, herpesvirus), cytomegalovirus (CMV,
herpesvirus) and Semliki forest virus (SFV, alphavirus). Compounds
1 and 2 inhibit infection of each of these various unrelated
enveloped viruses with approximately the same IC.sub.50.
[0308] These compounds are generally effective against all
lipid-enveloped viruses. The mechanism is expected to be the same
as the one of rhodanine compounds we previously described [1]. In
particular, we showed that this class of compounds (a) act on the
virus at a much lower concentration than the one toxic for the host
cells (FIG. 1 and FIG. 2), (b) irreversibly bind to the lipid
membrane of enveloped viruses, (c) disrupt the integrity of
lipid-enveloped viruses, and (d) renders treated viruses
non-infectious under inhibitory concentrations that are
non-cytotoxic. The mechanism of action is as follows: the non-polar
(hydrophobic) right-hand part of the oxazolidinedithiones localizes
the molecule preferentially in the non-polar hydrophobic lipid
membrane environment; the incidence of normal ambient light
photochemically activates the oxazolidinedithione molecule and
induces formation of singlet oxygen from cellular oxygen which is
likely to be concentrated in the lipid environment as well; the
generation of singlet oxygen in the vicinity of the hydrophobic
lipid membrane causes oxidative damage to the hydrophobic lipid
chain and thereby disrupts the role of the lipid membrane through
some as-yet unknown mechanism (destruction of the lipid chains,
change in curvature of the membrane, change in polarity of the
membrane, etc.).
[0309] Compound 1 shows remarkable inhibition of the infection of
lipid-enveloped viruses, as previously mentioned, and at lower
concentrations (IC.sub.50 .about.0.04 .mu.M) than the previously
described/claimed rhodanine derivatives [1]. Compound 2 shows even
higher antiviral activity (roughly ten times more active than 1,
IC.sub.50 .about.0.005 .mu.M). Therefore both of these compounds
(and perhaps their analogues) offer the potential for a better drug
for the broad-spectrum treatment of viral infections, a potential
antiviral topical or surface microbicide, as well as agents for
pathogen inactivation in transfusion medicine. In addition, due to
their specificity for lipid enveloped viruses, empiric response to
the viral inhibitory properties of active compound 1 and its
derivatives, can be used in the laboratory or clinical differential
diagnosis of lipid-enveloped versus non-enveloped viruses.
[0310] Broad spectrum antivirals are rare. For example, ribavarin
has activity against a few RNA virus by targeting RNA polymerase
function [2], but it is not acutely virucidal (i.e. it does not
kill the virus) and its efficacy varies tremendously when it does
work. Furthermore, resistance can develop easily to almost any
antiviral targeted against a viral protein. The in vitro IC.sub.50
(inhibitory efficacy) of compound 1 is relatively similar for all
the viruses tested, suggesting a common mechanism of action (as in
targeting the viral lipid membrane). In addition, since compound 1
effectively targets the viral lipid membrane and not a viral
protein, it is difficult for the virus to develop resistance even
in principle. Moreover, since the effects target the lipid membrane
rather than the genome (DNA or RNA) no mutagenic effect is
expected. Finally, unlike many other photosensitizers that
necessitate UV to be active, these compounds absorb in the visible
light range spectrum, thus reducing the hazard associated with the
use of UV light.
[0311] We previously carried out a wide-ranging array of biological
assays that showed the potent antiviral activity of the rhodanine
derivative that acts as a photosensitizer in the same way than the
compounds herein described [1]. We have tested the previous
compounds against enveloped and non-enveloped viruses encompassing
several taxonomic orders, families, genera, and species. We have
categorized the general mechanism of the compound's virucidal
activity as functioning in modification or disruption of the
enveloped virus' membrane, resulting in an irreversible inhibitory
effect on the virus particle. In vitro toxicity assays indicate a
relatively high therapeutic index with low cellular toxicity levels
at antiviral concentrations. We also tested the toxicity, efficacy,
and pharmacokinetics of the compounds in vivo in mouse dosing and
viral challenge models. The first mouse challenge experiment
indicated that the previous compound protected 100% of mice from
lethal Rift Valley Fever Virus challenge compared to the vehicle
(DMSO) control and an inactive analog (n=5 in each group). See [1]
for details. We now showed that: (1) the compound 1 and its
derivatives (compound 2 and other derivatives) are more effective
than the rhodanine derivatives; (2) antioxidants can protect from
the effects of the generated of singlet oxygen; (3) this class of
photosensitizers can show high cytotoxicity at low concentrations
upon light exposure, toxicity increasing with the time of exposure.
We are evaluating whether chemiluminescence can be used to activate
these compounds in a dark environment, and in vivo models of
application (antiviral). For example, association of the
oxazolidinedithiones with a system for local cellular light
generation, e.g., coupling with an enzyme such as luciferase and
then addition of luciferin, might provide the energy necessary to
generate singlet oxygen intracellularly and thereby produce in vivo
antiviral activity. Alternatively, organic nanocrystals (sodium
yttrium fluoride based) have been developed that are capable of
near-infrared to visible upconversion fluorescence. Such
nanocrystals have been coated with photosensitizers and activation
by tissue-penetrating near IR wavelengths can lead to singlet
oxygen generation and apoptosis of cancer cells targeted with these
nanocrystals (Guo H et al, Nanomedicine. 2010 Jan. 4. [Epub ahead
of print]; Bechet et al "Nanoparticles as vehicles for delivery of
photodynamic therapy agents" Trends in Biotechnology, 26:612-21).
If our new series of active oxazolidine-2,4-dithiones can be
conjugated to these upconverting nanocrystals, tissue penetrating
near-IR light could be used to control activation of our antiviral
in vivo.
[0312] The specific antiviral activity of compounds 1 and 2
involves their ability to interact with and disrupt the membrane
integrity of all lipid-enveloped viruses tested to date. Several
uses are possible. For example, the active compound(s) and its
derivatives can be used for therapeutic treatment of active viral
infections (with lipid-enveloped viruses). This can include acute
or chronic treatment with the compound depending on the pathogenic
profile of the virus. The compound(s) and its derivatives can also
be used for differential diagnosis between lipid enveloped versus
non-enveloped viruses in an in vitro or in vivo setting against
known or unknown viruses. For example, inhibition of virus
infection or virus induced cytopathic effect with the compound will
indicate that the pathogen in question is a lipid-enveloped virus,
and will help guide further diagnostic steps. The active
compound(s) and its derivatives can also be used for a topical
microbicide that might be effective against mucosally transmitted
lipid enveloped viruses such as HIV and Herpes Simplex Virus (HSV-1
and -2). The active compound(s) and its derivatives can also be
used as sterilizing agents or decontaminants of blood products. The
active compound(s) and its derivatives can also be used as
sterilizing agents or decontaminants of surfaces.
[0313] We have also shown that compounds 1 and 2 (and perhaps their
analogues) are more effective against HIV, NDV and HSV than
rhodanine derivatives, with IC.sub.50's in the nanomolar range (see
Table 1, "IC.sub.50's"). Other disclosed compounds similarly show
improved efficacy against HIV, NDV and/or HSV (see also Table 1).
In addition, we have also shown that compounds 1 and 2 (and perhaps
their analogues) generate more singlet oxygen (increased quantum
yield) than rhodanine derivatives (see Table 1, "1O2 QY"). Other
disclosed compounds similarly demonstrate improved generation of
singlet oxygen.
TABLE-US-00005 TABLE 1 Relative integrated IC50 .+-. STD (nM) Abs
area 1O2 QY (N = x) Max from 400 532 355 KT .times. 10.sup.-7 name
Structure HIV HSV NDV (nm) to 700 nm nm nm (M.sup.-1s.sup.-1) JL101
##STR00039## >1,000 (2) >1,000 (3) JL102 ##STR00040## 60 .+-.
63 (5) 2.8 .+-. 1.8 (3) 35 .+-. 17 (6) 501 134 0.80 .+-. 0.05 0.53
.+-. 0.04 16.0 .+-. 0.8 JL103 ##STR00041## 3 .+-. 0 (1) 2.3 .+-.
2.7 (4) 7 .+-. 7 (11) 516 153 0.63 .+-. 0.03 0.48 .+-. 0.02 7.8
.+-. 0.6 JL104 ##STR00042## Not active >500,000 (1) >500,000
(2) JL105 ##STR00043## >100,000 (3) 521 JL107 ##STR00044## 928
.+-. 432 (2) JL108 ##STR00045## 5 .+-. 3 (4) 526 188 0.52 .+-. 0.03
0.53 .+-. 0.03 4.9 .+-. 0.4 JL109 ##STR00046## 0.8 .+-. 0.6 (2) 4
.+-. 4 (6) 550 178 0.38 .+-. 0.02 0.37 .+-. 0.02 2.0 .+-. 0.1 JL110
##STR00047## 127 .+-. 101 (4) JL111 ##STR00048## 12 .+-. 1 (3) 499
JL113 ##STR00049## 865 .+-. 519 (2) JL117 ##STR00050## 20 .+-. 0
(1) 5 .+-. 3 (2) 504 JL118 ##STR00051## 2 .+-. 1 (4) 606 200 0.15
.+-. 0.02 0.14 .+-. 0.01 4.6 .+-. 0.3 JL119 ##STR00052## 6 .+-. 2
(3) 551 JL121 ##STR00053## Not active (1) >500,000 (2) 0 0 2.5
.+-. 0.2 JL122 ##STR00054## 3 .+-. 1 (2) 1 (1) 547 JL123
##STR00055## 75 .+-. 45 (2) 49 (1) 593 LJ001 ##STR00056## 183 .+-.
272 (9) 35 .+-. 48 (3) 207 .+-. 110 (10) 457 100 0 0.35 .+-. 0.03
1.0 .+-. 0.1 LJ025 ##STR00057## Not active (4) Not active (1) Not
active (2) 405
[0314] Notably, compounds 1 and 2 (and perhaps their analogues
exhibit) less toxicity and higher bioavailability in vivo, when
compared to the rhodanine derivatives, after either gavage or
intravenous or intraperitoneal administration in mice.
[0315] The pharmacokinetic (PK) profile of LJ-001, JL-102 and
JL-103 was determined after a single oral gavage (po),
intraperitoneal (ip) or intravenous (iv) administration to female
Balb/c mice.
[0316] Mice were treated with a single dose of the 3 test articles
at 10 mg/kg iv, ip and po, and 100 mg/kg po. In the 3 iv dose
groups, clinical signs, including tremors, hypoactivity or rough
fur, were observed in some of the mice. Out of the first 3 mice
treated with LJ-001 at 10 mg/kg ip, 2 were sacrificed in moribund
condition and 1 was found dead shortly after dose administration.
These mice were replaced and the dose level was reduced to 5 mg/kg.
Mice treated with LJ-001 at 5 mg/kg ip, and JL-102 and JL103 at 10
mg/kg ip appeared normal. Hypoactivity was the only clinical
observation noted in some of the mice treated with the 3 test
articles at 10 and 100 mg/kg po.
[0317] LJ-001 (10 mg/kg iv, 5 mg/kg ip, 10 and 100 mg/kg po) showed
minimal exposure due in part to a very short half-life (0.2 hr) and
fast elimination (.about.42000 ml/hr/kg). Oral bioavailability was
not determined as there was only one plasma data point in each po
dose group. JL-102 (10 mg/kg iv, 10 mg/kg ip, 10 and 100 mg/kg po)
and JL-103 (10 mg/kg iv, 10 mg/kg ip, 10 and 100 mg/kg po) had
comparable plasma time courses but the PK parameters showed
differences specific to dose route and level. The 10 mg/kg po
groups for JL-102 and JL-103 showed unexpectedly lower exposure
than the 100 mg/kg po groups, possibly due to significant
precipitation in the GI tract for the high dose levels. The ip
bioavailability for JL-102 and JL-103 was near 50% but the po
bioavailability ranged from 3.1 to 104.7%. The three drugs
distributed well into tissues.
EXAMPLE 4
Inhibition of Antiviral Activity by Antioxidants
[0318] It has been observed that antioxidants (e.g.,
alpha-tocopherol) offer protection from the antiviral effects of
the generated singlet oxygen. The ability of the antioxidant
alpha-tocopherol (a form of vitamin E) to protect viruses from the
antiviral activity of compound 2 was evaluated using NDV virus. NDV
was preincubated with increasing concentrations of alpha-tocopherol
and then treated with 15 nM EC.sub.90) of compound 2, which
maintains its ability to infect cells even when the appropriate
amount of alpha-tocopherol is provided (sec FIG. 3).
EXAMPLE 5
HIV Inactivation in Packed Red Blood Cells
[0319] The disclosed compounds, including compound 2 (and perhaps
analogues thereof) can be used to decontaminate blood products
containing red blood cells (RBC) at various hematocrit (Hct) levels
(see FIG. 4, HTV inactivation at a Hct of 80% by various selected
compounds). Note that Hct 50% and 80% mimick RBC concentration in
whole blood and in standard donor blood units, respectively.
[0320] Expired red blood cells (RBC) from the blood bank were
spiked with HIV at a final hematocrit (Hct) of approximately 80%
and treated with 100 .mu.M of selected compounds for 1 h under
white light on a shaker at 120 rpm. Cells were then centrifuged and
the supernatant used to infect reporter TZM-BL cells. At 48 h
post-infection, cells were lysed and the luciferase activity in the
lysate measured (see FIG. 4). While LJ001 is unable to inactivate
HIV at this concentration in an 80% Hct, some derivatives of
compound 2 are able to inactivate HIV. Indeed, the compounds JL109,
JL118 and JL122 all dramatically decrease the luciferase activity
to near-background levels, indicating a dramatic reduction of
infectious HIV particles in the packed RBC. Of note, these
compounds also appear more effective in these conditions than the
well-described and commonly used photosensitizers Hypericin and
Methylene Blue (MB). Without being bound by a particular theory, it
is believed that the effective compounds absorb at wavelengths that
are higher than the one of the hemoglobin contained in red blood
cells and thus can be excited by the light provided and generate
singlet oxygen, triggering the inactivation of the virus. However,
other mechanisms may be implied as both Hypericin and MB also have
absorption peaks at wavelengths higher than 600 nm.
EXAMPLE 6
Anti-Parasitic Activity Tested with Toxoplasma gondii
[0321] Attachment and invasion were assessed as described by Wetzel
D M, et al. (J Cell Sci. 2004 Nov. 15; 117(Pt 24):5739-48. Epub
2004 Oct. 26.). Parasites were pelleted at 2500 rpm in a tabletop
clinical centrifuge and washed in PBS. Equal numbers of parasites
were resuspended in DMEM and compounds added to 10 .mu.M with the
DMSO control at 1%. Parasites were incubated in the presence of
drug on ice for 15 min, then allowed to settle on coverslips of
confluent HFFs on ice for 15 min. After being allowed to settle the
cold media was changed out for warm media and parasites were
allowed to invade at 37.degree. C. for 1 hour, after which,
coverslips were rinsed with ice cold PBS and fixed. Two color
staining to discriminate between intracellular and extracellular
parasites was as described by Straub, K. W., et al. (PLoS Pathog.
2011 March; 7(3):e1002007. Epub 2011 Mar. 10) Five random fields
were counted on each coverslip and each experiment was done in
triplicate. Results were normalized to DMSO control.
[0322] JL102, 103 and 107 inhibited parasite attachment and
invasion with similar efficiency as LJ001 although variations in
efficacy were observed. Preliminary results suggest JL107 has most
potent inhibition of parasite attachment and invasion.
REFERENCES
[0323] [1] Wolf, M. C.; Freiberg, A. N.; Zhang, T.; Akyol-Ataman,
Z.; Grock, A.; Hong, P. W.; Li, J.; Watson, N. F.; Fang, A. Q.;
Aguilar, H. C.; Porotto, M.; Honko, A. N.; Damoiseaux, R.; Miller,
J. P.; Woodson, S. E.; Chantasirivisal, S.; Fontanes, V.; Negrete,
O. A.; Krogstad, P.; Dasgupta, A.; Moscona, A.; Hensley, L. E.;
Whelan, S. P.; Faull, K. F.; Holbrook, M. R.; Jung, M. E.; Lee, B.
A broad-spectrum antiviral targeting entry of enveloped viruses.
Proc Natl Acad Sci USA, 2010, 7, 3157-62.
[0324] [2] Parker, W. B. Metabolism and antiviral activity of
ribavirin. Virus Res, 2005, 2, 165-71.
[0325] The description of the aspects of the invention has been
presented for the purpose of illustration; it is not intended to be
exhaustive or to limit the invention to the precise forms
disclosed. Persons skilled in the relevant art can appreciate that
many modifications and variations are possible in light of the
above teachings. It should be noted that the language used in the
specification has been principally selected for readability and
instructional purposes, and it may not have been selected to
delineate or circumscribe the inventive subject matter.
Accordingly, the disclosure of the aspects of the invention is
intended to be illustrative, but not limiting, of the scope of the
invention.
[0326] Any terms not directly defined herein shall be understood to
have the meanings commonly associated with them as understood
within the art of the invention. Certain terms are discussed herein
to provide additional guidance to the practitioner in describing
the compositions, devices, methods and the like of aspects of the
invention, and how to make or use them. It will be appreciated that
the same thing can be said in more than one way. Consequently,
alternative language and synonyms can be used for any one or more
of the terms discussed herein. No significance is to be placed upon
whether or not a term is elaborated or discussed herein. Some
synonyms or substitutable methods, materials and the like are
provided. Recital of one or a few synonyms or equivalents does not
exclude use of other synonyms or equivalents, unless it is
explicitly stated. Use of examples, including examples of terms, is
for illustrative purposes only and does not limit the scope and
meaning of the aspects of the invention herein.
[0327] It must be noted that, as used in the specification, the
singular forms "a," "an" and "the" include plural referents unless
the context clearly dictates otherwise.
[0328] All publications, references, issued patents and patent
applications cited within the body of the specification are hereby
incorporated by reference in their entirety, for all purposes.
* * * * *