U.S. patent application number 15/148241 was filed with the patent office on 2016-09-01 for antiviral drugs for treatment or prevention of dengue infection.
This patent application is currently assigned to Siga Technologies, Inc.. The applicant listed for this patent is Siga Technologies, Inc.. Invention is credited to Tove BOLKEN, Dongcheng DAI, Dennis E. HRUBY, Robert JORDAN, Chelsea OLSEN.
Application Number | 20160250233 15/148241 |
Document ID | / |
Family ID | 40075507 |
Filed Date | 2016-09-01 |
United States Patent
Application |
20160250233 |
Kind Code |
A1 |
OLSEN; Chelsea ; et
al. |
September 1, 2016 |
ANTIVIRAL DRUGS FOR TREATMENT OR PREVENTION OF DENGUE INFECTION
Abstract
Compounds, methods and pharmaceutical compositions for treating
viral infections, by administering certain compounds in
therapeutically effective amounts are disclosed. Methods for
preparing the compounds and methods of using the compounds and
pharmaceutical compositions thereof are also disclosed. In
particular, the treatment and prophylaxis of viral infections such
as caused by flavivirus is disclosed, i.e., including but not
limited to, Dengue virus, West Nile virus, yellow fever virus,
Japanese encephalitis virus, and tick-borne encephalitis virus.
Inventors: |
OLSEN; Chelsea; (Kirkland,
WA) ; JORDAN; Robert; (Foster City, CA) ; DAI;
Dongcheng; (Corvallis, OR) ; BOLKEN; Tove;
(Keizer, OR) ; HRUBY; Dennis E.; (Albany,
OR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Siga Technologies, Inc. |
Corvallis |
OR |
US |
|
|
Assignee: |
Siga Technologies, Inc.
Corvallis
OR
|
Family ID: |
40075507 |
Appl. No.: |
15/148241 |
Filed: |
May 6, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13947630 |
Jul 22, 2013 |
9353051 |
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15148241 |
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12601187 |
Apr 5, 2010 |
8518960 |
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PCT/US2008/064662 |
May 23, 2008 |
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13947630 |
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60924628 |
May 23, 2007 |
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Current U.S.
Class: |
514/157 |
Current CPC
Class: |
A61P 31/12 20180101;
Y02A 50/395 20180101; A61K 31/517 20130101; C07C 311/46 20130101;
C07D 409/12 20130101; A61K 31/502 20130101; A61K 31/522 20130101;
A61P 31/14 20180101; C07D 401/12 20130101; C07D 221/14 20130101;
A61K 31/635 20130101; C07D 241/44 20130101; C07D 491/048 20130101;
A61K 31/519 20130101; Y02A 50/389 20180101; A61K 31/473 20130101;
C07D 253/08 20130101; Y02A 50/393 20180101; C07D 403/06 20130101;
Y02A 50/385 20180101; A61K 31/63 20130101; C07D 239/88 20130101;
Y02A 50/387 20180101; C07D 473/08 20130101; C07D 417/12 20130101;
A61K 31/18 20130101; A61K 31/53 20130101; C07D 237/32 20130101;
C07D 239/96 20130101; C07D 495/04 20130101; A61K 9/0019 20130101;
C07D 405/12 20130101 |
International
Class: |
A61K 31/635 20060101
A61K031/635; A61K 31/18 20060101 A61K031/18; A61K 31/53 20060101
A61K031/53; A61K 31/517 20060101 A61K031/517; A61K 31/63 20060101
A61K031/63; A61K 9/00 20060101 A61K009/00 |
Claims
1.-36. (canceled)
37. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and a compound having the following general
formula I or a pharmaceutically acceptable salt thereof:
##STR00161## wherein R.sup.1 and R.sup.2 are independently
hydrogen, alkyl, alkenyl, alkynyl, or unsubstituted or substituted
cycloalkyl, arylalkyl, aryl, or R.sup.1 and R.sup.2 together may
form a substituted or unsubstituted ring, which may include one or
more heteroatoms in the ring; and Ar is substituted or
unsubstituted aryl or heteroaryl; said cycloalkyl, arylalkyl, and
aryl group substituents being one or more radical(s) independently
selected from the group consisting of a straight- or branched chain
alkyl, alkoxy, alkoxyalkyl, alkoxyalkoxy, halogen, polyfluoroalkyl,
polyfluoroalkoxy, carboxy, cyano, nitro, amido, amidoalkyl,
carboxamide, alkylthio, alkylsulfinyl, alkylsulfonyl, sulfonamide,
and mercapto wherein the compound of formula I is selected from
2-(6-Chloro-4-oxo-4H-quinazolin-3-yl)-N4-[4-(isopropyl-methyl-sulfamoyl)--
phenyl]-acetamide;
2-(7-Fluoro-4-oxo-4H-quinazolin-3-yl)-N-[4-(isopropyl-methyl-sulfamoyl)-p-
henyl]-acetamide;
N-[4-(2,6-Dimethyl-piperidine-1-sulfonyl)-phenyl]-2-(4-oxo-4H-quinazolin--
3-yl)-acetamide;
N-[4-(3,5-Dimethyl-piperidine-1-sulfonyl)-phenyl]-2-(4-oxo-4H-quinazolin--
3-yl)-acetamide;
N-[4-(2,6-Dimethyl-piperidine-1-sulfonyl)-phenyl]-2-(4-oxo-4H-thieno[2,3--
d]pyrimidin-3-yl)-acetamide; and
2-(4-oxo-4H-quinazolin-3-yl)-N-[4-(2,2,2-trifluoro-ethylsulfamoyl)-phenyl-
]-acetamide.
38. The pharmaceutical composition according to claim 37, wherein
said pharmaceutical composition is suitable for administration in a
human or animal, wherein said administration is selected from the
group consisting of: oral administration, rectal administration,
parenteral administration, intravaginal administration,
intraperitoneal administration and administration by
inhalation.
39. The pharmaceutical composition according to claim 37, wherein
said pharmaceutically acceptable carrier is solid.
40. The pharmaceutical composition according to claim 37, wherein
said pharmaceutically acceptable carrier is liquid.
41. The pharmaceutical composition according to claim 38, wherein
said parenteral administration is selected from the group
consisting of: intramuscular injection, subcutaneous injection and
intravenous infusion.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to and benefit of U.S.
Provisional Application No. 60/924,628, filed May 23, 2007, the
contents of which are hereby incorporated by reference in their
entirety.
FIELD OF THE INVENTION
[0002] This invention relates to the use of benzenesulfonamide
derivatives and analogs, as well as compositions containing the
same, for the treatment or prophylaxis of viral diseases associated
with the flavivirus family such as Dengue fever, Yellow fever, West
Nile, St. Louis encephalitis, Hepatitis C, Murray Valley
encephalitis, and Japanese encephalitis.
BACKGROUND OF THE INVENTION
[0003] Dengue fever (DF) is an acute febrile disease caused by one
of four closely related virus serotypes (DEN-1, DEN-2, DEN-3, and
DEN-4). Dengue fever is classified based on its clinical
characteristics into classical dengue fever, or the more severe
forms, dengue hemorrhagic fever syndrome (DHF), and dengue shock
syndrome (DSS). Recovery from infection from one serotype produces
life-long immunity to that particular serotype, but provides only
short-lived and limited protection against any of the other
serotypes (37). Dengue is a member of the Flaviviridae family which
are enveloped, positive-sense RNA viruses whose human pathogens
also include West Nile virus (WNV), yellow fever virus (YFV),
Japanese encephalitis virus (JEV), and tick-borne encephalitis
virus (TBEV) among others. Dengue transmission is via the bite of
an infected Aedes aegypti mosquito which is found in tropical and
sub-tropical regions around the world.
[0004] Each year regional epidemics of dengue cause significant
morbidity and mortality, social disruption and substantial economic
burden on the societies affected both in terms of hospitalization
and mosquito control. Dengue is considered by the World Health
Organization (WHO) to be the most important arthropod-borne viral
disease with an estimated 50 million cases of dengue infection,
including 500,000 DHF cases and 24,000 deaths worldwide each year
(37, 38). WHO estimates that forty percent of the world's
population (2.5 billion people) are at risk for DF, DHF, and DSS
(37). Dengue is also a NIAID Category A pathogen and in terms of
bio-defense, represents a significant threat to United States
troops overseas. Dengue is an emerging threat to North America with
a dramatic increase in severe disease in the past 25 years
including major epidemics in Cuba and Venezuela, and outbreaks in
Texas and Hawaii (4). Failure to control the mosquito vector and
increases in long-distance travel have contributed to the increase
and spread of dengue disease. The characteristics of dengue as a
viral hemorrhagic fever virus (arthropod-borne, widely spread, and
capable of inducing a great amount of cellular damage and eliciting
an immune response that can result in severe hemorrhage, shock, and
death) makes this virus a unique threat to deployed military
personnel around the world as well as to travelers to tropical
regions. Preparedness for both biodefense and for the public health
challenges posed by dengue will require the development of new
vaccines and antiviral therapeutics.
[0005] Dengue causes several illnesses with increasing severity
being determined in part by prior infection with a different
serotype of the virus. Classic dengue fever (DF) begins 3-8 days
after the bite of an infected mosquito and is characterized by
sudden onset of fever, headache, back pain, joint pain, a
measles-like rash, and nausea and vomiting (21). DF is frequently
referred to as "breakbone" fever due to these symptoms. The disease
usually resolves after two weeks but a prolonged recovery with
weakness and depression is common. The more severe form of the
disease, dengue hemorrhagic fever (DHF) has a similar onset and
early phase of illness as dengue fever. However, shortly after
onset the disease is characterized by high fever, enlargement of
the liver, and hemorrhagic phenomena such as bleeding from the
nose, mouth, and internal organs due to vascular permeability (38).
In dengue shock syndrome (DSS) circulatory failure and hypovolaemic
shock resulting from plasma leakage occur and can lead to death in
12-24 hours without plasma replacement (38). The case fatality rate
of DHF/DSS can be as high as 20% without treatment. DHF has become
a leading cause of hospitalization and death among children in many
countries with an estimated 500,000 cases requiring hospitalization
each year and a case fatality rate of about 5%(37).
[0006] The pathogenesis of DHF/DSS is still being studied but is
thought to be due in part to an enhancement of virus replication in
macrophages by heterotypic antibodies, termed antibody-dependent
enhancement (ADE) (8). During a secondary infection, with a
different serotype of dengue virus, cross-reactive antibodies that
are not neutralizing form virus-antibody complexes that are taken
into monocytes and Langerhans cells (dendritic cells) and increase
the number of infected cells (7). This leads to the activation of
cytotoxic lymphocytes which can result in plasma leakage and the
hemorrhagic features characteristic of DHF and DSS (21). This
antibody-dependent enhancement of infection is one reason why the
development of a successful vaccine has proven to be so difficult.
Although less frequent, DHF/DSS can occur after primary infection
(33), so virus virulence (16) and immune activation are also
believed to contribute to the pathogenesis of the disease (26).
[0007] Dengue is endemic in more than 100 countries in Africa, the
Americas, the Eastern Mediterranean, South-east Asia and the
Western Pacific. During epidemics, attack rates can be as high as
80-90% of the susceptible population. All four serotypes of the
virus are emerging worldwide, increasing the number of cases of the
disease as well as the number of explosive outbreaks. In 2002 for
example, there were 1,015,420 reported cases of dengue in the
Americas alone with 14,374 cases of DHF, which is more than three
times the number of dengue cases reported in the Americas in 1995
(24).
[0008] The dengue genome, approximately 11 kb in length, consists
of a linear, single stranded, infectious, positive sense RNA that
is translated as a single long polyprotein (reviewed in (29). The
genome is composed of seven nonstructural (NS) protein genes and
three structural protein genes which encode the nucleocapsid
protein (C), a membrane-associated protein (M), and an envelope
protein (E). The nonstructural proteins are involved in viral RNA
replication (35), viral assembly, and the inflammatory components
of the disease (19). The structural proteins are involved mainly in
viral particle formation (22). The precursor polyprotein is cleaved
by cellular proteinases to separate the structural proteins (18),
while a virus-encoded proteinase cleaves the nonstructural region
of the polyprotein (6). The genome is capped and does not have a
poly(A) tail at the 3' end but instead has a stable stem-loop
structure necessary for stability and replication of the genomic
RNA (3). The virus binds to cellular receptors via the E protein
and undergoes receptor-mediated endocytosis followed by low-pH
fusion in lysosomes (20). The viral genome is then uncoated and
translated into the viral precursor polyprotein. Co- and
posttranslational proteolytic processing separates the structural
and nonstructural proteins. The RNA-dependent RNA polymerase along
with cofactors synthesizes the minus-strand RNA which serves as a
template for the synthesis of the progeny plus-strand RNA (25).
Viral replication is membrane associated (1, 34). Following
replication, the genome is encapsidated, and the immature virus,
surrounded by a lipid envelope buds into the lumen (9). The
envelope proteins become glycosylated and mature viruses are
released outside the cell. Essential stages or process during the
virus life cycle would be possible targets for inhibition from an
antiviral drug and include binding of the virus to the cell through
the E protein, uptake of the virus into the cell, the capping
mechanism, the viral proteinase, the viral RNA-dependent RNA
polymerase, and the viral helicase.
[0009] Current management of dengue virus-related disease relies
solely on vector control. There are no approved antivirals or
vaccines for the treatment or prevention of dengue. Ribavirin, a
guanosine analogue, has been shown to be effective against a range
of RNA virus infections and works against dengue in tissue culture
by inhibiting the dengue 2'-O-methyltransferase NS5 domain (2, 10).
However, ribavirin did not show protection against dengue in a
mouse model (15) or a rhesus monkey model (17), instead it induced
anemia and thrombocytosis. While there are no currently available
approved vaccines, multivalent dengue vaccines have shown some
limited potential in humans (5, 12, 13, 28). However, vaccine
development is difficult due to the presence of four distinct
serotypes of the virus which each cause disease. Vaccine
development also faces the challenge of ADE where unequal
protection against the different strains of the virus could
actually increase the risk of more serious disease. Therefore there
is a need for antiviral drugs that target all of the serotypes of
dengue. An antiviral drug administered early during dengue
infection that inhibits viral replication would prevent the high
viral load associated with DHF and be an attractive strategy in the
treatment and prevention of disease. An antiviral drug that
inhibits viral replication could be administered prior to travel to
a dengue endemic region to prevent acquisition of disease, or for
those that have previously been exposed to dengue, could prevent
infection by another serotype of virus and decrease the chance of
life-threatening DHF and DSS. Having an antiviral drug would also
aid vaccine development by having a tool at hand to treat
complications that may arise due to unequal immune protection
against the different serotypes. Although a successful vaccine
could be a critical component of an effective biodefense, the
typical delay to onset of immunity, potential side-effects, cost,
and logistics associated with large-scale civilian vaccinations
against a low-threat risk agent suggest that a comprehensive
biodefense include a separate rapid-response element. Thus, there
remains an urgent need to develop a safe and effective product to
protect against flavivirus infection.
SUMMARY OF THE INVENTION
[0010] The present invention provides a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound
having the following general formula I or a pharmaceutically
acceptable salt thereof:
##STR00001##
[0011] wherein R.sup.1 and R.sup.2 are independently hydrogen,
alkyl, alkenyl, alkynyl, or unsubstituted or substituted
cycloalkyl, arylalkyl, aryl, or R.sup.1 and R.sup.2 together may
form a substituted or unsubstituted ring, which may include one or
more heteroatoms in the ring; and
[0012] Ar is substituted or unsubstituted aryl or heteroaryl;
said cycloalkyl, arylalkyl, and aryl group substituents being one
or more radical(s) independently selected from the group consisting
of a straight- or branched chain alkyl, alkoxy, alkoxyalkyl,
alkoxyalkoxy, halogen, polyfluoroalkyl, polyfluoroalkoxy, carboxy,
cyano, nitro, amido, amidoalkyl, carboxamide, alkylthio,
alkylsulfinyl, alkylsulfonyl, sulfonamide, and mercapto.
[0013] The present invention also provides a method for the
treatment or prophylaxis of a viral infection or disease associated
therewith, comprising administering in a therapeutically effective
amount to a mammal in need thereof, a compound of Formula I below
or a pharmaceutically acceptable salt thereof:
##STR00002##
[0014] wherein R.sup.1 and R.sup.2 are independently hydrogen,
alkyl, alkenyl, alkynyl, or unsubstituted or substituted
cycloalkyl, arylalkyl, aryl, or R.sup.1 and R.sup.2 together may
form a substituted or unsubstituted ring, which may include one or
more heteroatoms in the ring; and
[0015] Ar is substituted or unsubstituted aryl or heteroaryl;
said cycloalkyl, arylalkyl, and aryl group substituents being one
or more radical(s) independently selected from the group consisting
of a straight- or branched chain alkyl, alkoxy, alkoxyalkyl,
alkoxyalkoxy, halogen, polyfluoroalkyl, polyfluoroalkoxy, carboxy,
cyano, nitro, amido, amidoalkyl, carboxamide, alkylthio,
alkylsulfinyl, alkylsulfonyl, sulfonamide, and mercapto.
[0016] Other objects and advantages of the present invention will
become apparent from the following description and appended
claims.
DETAILED DESCRIPTION OF THE INVENTION
[0017] The compounds of the invention are of the following general
Formula I:
##STR00003##
wherein R.sup.1 and R.sup.2 are independently hydrogen, alkyl,
alkenyl, alkynyl, or unsubstituted or substituted cycloalkyl,
arylalkyl, aryl, or R.sup.1 and R.sup.2 together may form a
substituted or unsubstituted ring, which may include one or more
heteroatoms in the ring; and
[0018] Ar is substituted or unsubstituted aryl or heteroaryl;
said cycloalkyl, arylalkyl, and aryl group substituents being one
or more radical(s) independently selected from the group consisting
of a straight- or branched chain alkyl, alkoxy, alkoxyalkyl,
alkoxyalkoxy, halogen, polyfluoroalkyl, polyfluoroalkoxy, carboxy,
cyano, nitro, amido, amidoalkyl, carboxamide, alkylthio,
alkylsulfinyl, alkylsulfonyl, sulfonamide, and mercapto.
[0019] Preferably each of R.sup.1 and R.sup.2 is an ethyl. Also
preferably, Ar is a mono-substituted phenyl such as amido-phenyl.
Examples of amido-phenyl include isobutyramidophenyl,
p-[2-(4-oxo-4H-quinazolin-3-yl)-acetamido]-phenyl and
p-[2-(4-oxo-4H-benzo[d][1,2,3]triazin-3-yl)-acetamido]-phenyl.
Again preferably, Ar is a di-substituted phenyl with one
substituent as amido and the other one as methoxy. Examples of this
type of phenyl include m-phenyl-acetamido-p-methoxy-phenyl and
m-(3-methyl-butyramido)-p-methoxy-phenyl.
[0020] Most preferably, the compound of Formula I is
N-(4-Diethylsulfamoyl-phenyl)-2-(4-oxo-4H-quinazolin-3-yl)-acetamide.
Exemplary compounds according to the invention are shown below in
Table 1.
[0021] The method of the present invention is for the treatment or
prophylaxis of a viral infection or disease associated therewith,
comprising administering in a therapeutically effective amount to a
mammal in need thereof, a compound of Formula I described
above.
[0022] Preferably, the mammal is a human and the viral infection is
a flavivirus infection. More preferably, the flavivirus virus is
selected from the group consisting of Dengue virus, West Nile
virus, yellow fever virus, Japanese encephalitis virus, and
tick-borne encephalitis virus. Most preferably, the flavivirus is a
Dengue virus selected from the group consisting of DEN-1, DEN-2,
DEN-3, and DEN-4.
[0023] Preferably, the viral infection is associated with a
condition selected from the group consisting of Dengue fever,
Yellow fever, West Nile, St. Louis encephalitis, Hepatitis C,
Murray Valley encephalitis, and Japanese encephalitis. Most
preferably, the viral infection is associated with Dengue fever
wherein said Dengue fever is selected from the group consisting of
classical dengue fever, dengue hemorrhagic fever syndrome, and
dengue shock syndrome.
[0024] The method of the present invention may also comprise
co-administration of: a) other antivirals such as Ribavirin or
cidofovir; b) vaccines; and/or c) interferons or pegylated
interferons.
DEFINITIONS
[0025] In accordance with this detailed description, the following
abbreviations and definitions apply. It must be noted that as used
herein, the singular forms "a," "an," and "the" include plural
referents unless the context clearly dictates otherwise.
[0026] The publications discussed herein are provided solely for
their disclosure. Nothing herein is to be construed as an admission
regarding antedating the publications. Further, the dates of
publication provided may be different from the actual publication
dates, which may need to be independently confirmed.
[0027] Where a range of values is provided, it is understood that
each intervening value is encompassed. The upper and lower limits
of these smaller ranges may independently be included in the
smaller, subject to any specifically-excluded limit in the stated
range. Where the stated range includes one or both of the limits,
ranges excluding either both of those included limits are also
included in the invention. Also contemplated are any values that
fall within the cited ranges.
[0028] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art. Any methods and materials similar or
equivalent to those described herein can also be used in practice
or testing. All publications mentioned herein are incorporated
herein by reference to disclose and describe the methods and/or
materials in connection with which the publications are cited.
[0029] By "patient" or "subject" is meant to include any mammal. A
"mammal," for purposes of treatment, refers to any animal
classified as a mammal, including but not limited to, humans,
experimental animals including rats, mice, and guinea pigs,
domestic and farm animals, and zoo, sports, or pet animals, such as
dogs, horses, cats, cows, and the like.
[0030] The term "efficacy" as used herein in the context of a
chronic dosage regime refers to the effectiveness of a particular
treatment regime. Efficacy can be measured based on change of the
course of the disease in response to an agent.
[0031] The term "success" as used herein in the context of a
chronic treatment regime refers to the effectiveness of a
particular treatment regime. This includes a balance of efficacy,
toxicity (e.g., side effects and patient tolerance of a formulation
or dosage unit), patient compliance, and the like. For a chronic
administration regime to be considered "successful" it must balance
different aspects of patient care and efficacy to produce a
favorable patient outcome.
[0032] The terms "treating," "treatment," and the like are used
herein to refer to obtaining a desired pharmacological and
physiological effect. The effect may be prophylactic in terms of
preventing or partially preventing a disease, symptom, or condition
thereof and/or may be therapeutic in terms of a partial or complete
cure of a disease, condition, symptom, or adverse effect attributed
to the disease. The term "treatment," as used herein, covers any
treatment of a disease in a mammal, such as a human, and includes:
(a) preventing the disease from occurring in a subject which may be
predisposed to the disease but has not yet been diagnosed as having
it, i.e., causing the clinical symptoms of the disease not to
develop in a subject that may be predisposed to the disease but
does not yet experience or display symptoms of the disease; (b)
inhibiting the disease, i.e., arresting or reducing the development
of the disease or its clinical symptoms; and (c) relieving the
disease, i.e., causing regression of the disease and/or its
symptoms or conditions. Treating a patient's suffering from disease
related to pathological inflammation is contemplated. Preventing,
inhibiting, or relieving adverse effects attributed to pathological
inflammation over long periods of time and/or are such caused by
the physiological responses to inappropriate inflammation present
in a biological system over long periods of time are also
contemplated.
[0033] "Alkenyl" refers to alkenyl group preferably having from 2
to 10 carbon atoms and more preferably 2 to 6 carbon atoms and
having at least 1 and preferably from 1-2 sites of alkenyl
unsaturation.
[0034] "Alkoxy" refers to the group "alkyl-O-" which includes, by
way of example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy,
tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy,
and the like.
[0035] "Alkyl" refers to linear or branched alkyl groups having
from 1 to 10 carbon atoms, alternatively 1 to 6 carbon atoms. This
term is exemplified by groups such as methyl, t-butyl, n-heptyl,
octyl and the like. "Amino" refers to the group --NH.sub.2.
[0036] "Aryl" or "Ar" refers to an unsaturated aromatic carbocyclic
group of from 6 to 14 carbon atoms having a single ring (e.g.,
phenyl) or multiple condensed rings (e.g., naphthyl or anthryl)
which condensed rings may or may not be aromatic (e.g.,
2-benzoxazolinone, 2H-1,4-benzoxazin-3(4H)-one, and the like)
provided that the point of attachment is through an aromatic ring
atom. "Substituted aryl" refers to aryl groups which are
substituted with from 1 to 3 substituents selected from the group
consisting of hydroxy, acyl, acylamino, thiocarbonylamino, acyloxy,
alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl, amidino,
alkylamidino, thioamidino, amino, aminoacyl, aminocarbonyloxy,
aminocarbonylamino, aminothiocarbonylamino, aryl, substituted aryl,
aryloxy, substituted aryloxy, cycloalkoxy, substituted cycloalkoxy,
heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy,
substituted heterocyclyloxy, carboxyl, carboxylalkyl,
carboxyl-substituted alkyl, carboxyl-cycloalkyl,
carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substituted
aryl, carboxylheteroaryl, carboxyl-substituted heteroaryl,
carboxylheterocyclic, carboxyl-substituted heterocyclic,
carboxylamido, cyano, thiol, thioalkyl, substituted thioalkyl,
thioaryl, substituted thioaryl, thioheteroaryl, substituted
thioheteroaryl, thiocycloalkyl, substituted thiocycloalkyl,
thioheterocyclic, substituted thioheterocyclic, cycloalkyl,
substituted cycloalkyl, guanidino, guanidinosulfone, halo, nitro,
heteroaryl, substituted heteroaryl, heterocyclic, substituted
heterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,
substituted heteroaryloxy, heterocyclyloxy, substituted
heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino,
--S(O).sub.2-alkyl, --S(O).sub.2-substituted alkyl,
--S(O).sub.2-cycloalkyl, --S(O).sub.2-substituted cycloalkyl,
--S(O).sub.2-alkenyl, --S(O).sub.2-substituted alkenyl,
--S(O).sub.2-aryl, --S(O).sub.2-substituted aryl,
--S(O).sub.2-heteroaryl, --S(O).sub.2-substituted heteroaryl,
--S(O).sub.2-heterocyclic, --S(O).sub.2-substituted heterocyclic,
--OS(O).sub.2-alkyl, --OS(O).sub.2-substituted alkyl,
--OS(O).sub.2-aryl, --OS(O).sub.2-substituted aryl,
--OS(O).sub.2-heteroaryl, --OS(O).sub.2-substituted heteroaryl,
--OS(O).sub.2-heterocyclic, --OS(O).sub.2-substituted heterocyclic,
--OS(O).sub.2--NRR where R is hydrogen. or alkyl,
--NRS(O).sub.2-alkyl, --NRS(O).sub.2-substituted alkyl,
--NRS(O).sub.2-aryl, --NRS(O).sub.2-substituted aryl,
--NRS(O).sub.2-heteroaryl, --NRS(O).sub.2-substituted heteroaryl,
--NRS(O).sub.2-heterocyclic, --NRS(O).sub.2-substituted
heterocyclic, --NRS(O).sub.2--NR-alkyl,
--NRS(O).sub.2--NR-substituted alkyl, --NRS(O).sub.2--NR-aryl,
--NRS(O).sub.2--NR-substituted aryl, --NRS(O).sub.2--NR-heteroaryl,
--NRS(O).sub.2--NR-substituted heteroaryl,
--NRS(O).sub.2--NR-heterocyclic, --NRS(O).sub.2--NR-substituted
heterocyclic where R is hydrogen or alkyl, mono- and di-alkylamino,
mono- and di-(substituted alkyl)amino, mono- and di-arylamino,
mono- and di-substituted arylamino, mono- and di-heteroarylamino,
mono- and di-substituted heteroarylamino, mono- and di-heterocyclic
amino, mono- and di-substituted heterocyclic amino, unsymmetric
di-substituted amines having different substituents independently
selected from the group consisting of alkyl, substituted alkyl,
aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic and amino groups on the
substituted aryl blocked by conventional blocking groups such as
Boc, Cbz, formyl, and the like or substituted with --SO.sub.2NRR
where R is hydrogen or alkyl.
[0037] "Cycloalkyl" refers to cyclic alkyl groups of from 3 to 8
carbon atoms having a single cyclic ring including, by way of
example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cyclooctyl and the like. Excluded from this definition are
multi-ring alkyl groups such as adamantanyl, etc.
[0038] "Halo" or "halogen" refers to fluoro, chloro, bromo and
iodo.
[0039] "Heteroaryl" refers to an aromatic carbocyclic group of from
2 to 10 carbon atoms and 1 to 4 heteroatoms selected from the group
consisting of oxygen, nitrogen and sulfur within the ring or oxides
thereof. Such heteroaryl groups can have a single ring (e.g.,
pyridyl or furyl) or multiple condensed rings (e.g., indolizinyl or
benzothienyl) wherein one or more of the condensed rings may or may
not be aromatic provided that the point of attachment is through an
aromatic ring atom. Additionally, the heteroatoms of the heteroaryl
group may be oxidized, i.e., to form pyridine N-oxides or
1,1-dioxo-1,2,5-thiadiazoles and the like. Additionally, the carbon
atoms of the ring may be substituted with an oxo (.dbd.O). The term
"heteroaryl having two nitrogen atoms in the heteroaryl, ring"
refers to a heteroaryl group having two, and only two, nitrogen
atoms in the heteroaryl ring and optionally containing 1 or 2 other
heteroatoms in the heteroaryl ring, such as oxygen or sulfur.
[0040] "Substituted heteroaryl" refers to heteroaryl groups which
are substituted with from 1 to 3 substituents selected from the
group consisting of hydroxy, acyl, acylamino, thiocarbonylamino,
acyloxy, alkyl, substituted alkyl, alkoxy, substituted alkoxy,
alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,
amidino, alkylamidino, thioamidino, amino, aminoacyl,
aminocarbonyloxy, aminocarbonylamino, aminothiocarbonylamino, aryl,
substituted aryl, aryloxy, substituted aryloxy, cycloalkoxy,
substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,
heterocyclyloxy, substituted heterocyclyloxy, carboxyl,
carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl,
carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substituted
aryl, carboxylheteroaryl, carboxyl-substituted heteroaryl,
carboxylheterocyclic, carboxyl-substituted heterocyclic,
carboxylamido, cyano, thiol, thioalkyl, substituted thioalkyl,
thioaryl, substituted thioaryl, thioheteroaryl, substituted
thioheteroaryl, thiocycloalkyl, substituted thiocycloalkyl,
thioheterocyclic, substituted thioheterocyclic, cycloalkyl,
substituted cycloalkyl, guanidino, guanidinosulfone, halo, nitro,
heteroaryl, substituted heteroaryl, heterocyclic, substituted
heterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,
substituted heteroaryloxy, heterocyclyloxy, substituted
heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino,
--S(O).sub.2-alkyl, --S(O).sub.2-substituted alkyl,
--S(O).sub.2-cycloalkyl, --S(O).sub.2-substituted cycloalkyl,
--S(O).sub.2-alkenyl, --S(O).sub.2-substituted alkenyl,
--S(O).sub.2-aryl, --S(O).sub.2-substituted aryl,
--S(O).sub.2-heteroaryl, --S(O).sub.2-substituted heteroaryl,
--S(O).sub.2-heterocyclic, --S(O).sub.2-substituted heterocyclic,
--OS(O).sub.2-alkyl, --OS(O).sub.2-substituted alkyl,
--OS(O).sub.2-aryl, --OS(O).sub.2-substituted aryl,
--OS(O).sub.2-heteroaryl, --OS(O).sub.2-substituted heteroaryl,
--OS(O).sub.2-heterocyclic, --OS(O).sub.2-substituted heterocyclic,
--OSO.sub.2--NRR where R is hydrogen or alkyl,
--NRS(O).sub.2-alkyl, --NRS(O).sub.2-substituted alkyl,
--NRS(O).sub.2-aryl, --NRS(O).sub.2-substituted aryl,
--NRS(O).sub.2-heteroaryl, --NRS(O).sub.2-substituted heteroaryl,
--NRS(O).sub.2-heterocyclic, --NRS(O).sub.2-substituted
heterocyclic, --NRS(O).sub.2--NR-alkyl,
--NRS(O).sub.2--NR-substiruted alkyl, --NRS(O).sub.2--NR-aryl,
--NRS(O).sub.2--NR-substituted aryl, --NRS(O).sub.2--NR-heteroaryl,
--NRS(O).sub.2--NR-substituted heteroaryl,
--NRS(O).sub.2--NR-heterocyclic, --NRS(O).sub.2--NR-substituted
heterocyclic where R is hydrogen or alkyl, mono- and di-alkylamino,
mono- and di-(substituted alkyl)amino, mono- and di-arylamino,
mono- and di-substituted arylamino, mono- and di-heteroarylamino,
mono- and di-substituted heteroarylamino, mono- and di-heterocyclic
amino, mono- and di-substituted heterocyclic amino, unsymmetric
di-substituted amines having different substituents independently
selected from the group consisting of alkyl, substituted alkyl,
aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic and amino groups on the
substituted aryl blocked by conventional blocking groups such as
Boc, Cbz, formyl, and the like or substituted with --SO.sub.2NRR
where R is hydrogen or alkyl.
[0041] "Sulfonyl" refers to the group --S(O).sub.2R where R is
selected from the group consisting of hydrogen, alkyl, substituted
alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,
aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,
heteroaryl, substituted heteroaryl, heterocyclic, substituted
heterocyclic wherein alkyl, substituted alkyl, alkenyl, substituted
alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted
cycloalkyl, aryl, substituted aryl, heteroaryl, substituted
heteroaryl, heterocyclic and substituted heterocyclic are as
defined herein.
[0042] "Pharmaceutically-acceptable carrier" means a carrier that
is useful in preparing a pharmaceutical composition or formulation
that is generally safe, non-toxic, and neither biologically nor
otherwise undesirable, and includes a carrier that is acceptable
for veterinary use as well as human pharmaceutical use. A
pharmaceutically-acceptable carrier or excipient includes both one
or more than one of such carriers.
[0043] "Pharmaceutically-acceptable cation" refers to the cation of
a pharmaceutically-acceptable salt. "Pharmaceutically-acceptable
salt" refers to salts which retain the biological effectiveness and
properties of compounds which are not biologically or otherwise
undesirable. Pharmaceutically-acceptable salts refer to
pharmaceutically-acceptable salts of the compounds, which salts are
derived from a variety of organic and inorganic counter ions well
known in the art and include, by way of example only, sodium,
potassium, calcium, magnesium, ammonium, tetraalkylammonium, and
the like; and when the molecule contains a basic functionality,
salts of organic or inorganic acids, such as hydrochloride,
hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and the
like.
[0044] Pharmaceutically-acceptable base addition salts can be
prepared from inorganic and organic bases. Salts derived from
inorganic bases, include by way of example only, sodium, potassium,
lithium, ammonium, calcium and magnesium salts. Salts derived from
organic bases include, but are not limited to, salts of primary,
secondary and tertiary amines, such as alkyl amines, dialkyl
amines, trialkyl amines, substituted alkyl amines, di(substituted
alkyl) amines, tri(substituted alkyl) amines, alkenyl amines,
dialkenyl amines, trialkenyl amines, substituted alkenyl amines,
di(substituted alkenyl) amines, tri(substituted alkenyl) amines,
cycloalkyl amines, di(cycloalkyl) amines, tri(cycloalkyl) amines,
substituted cycloalkyl amines, disubstituted cycloalkyl amine,
trisubstituted cycloalkyl amines, cycloalkenyl amines,
di(cycloalkenyl) amines, tri(cycloalkenyl) amines, substituted
cycloalkenyl amines, disubstituted cycloalkenyl amine,
trisubstituted cycloalkenyl amines, aryl amines, diaryl amines,
triaryl amines, heteroaryl amines, diheteroaryl amines,
triheteroaryl amines, heterocyclic amines, diheterocyclic amines,
triheterocyclic amines, mixed di- and tri-amines where at least two
of the substituents on the amine are different and are selected
from the group consisting of alkyl, substituted alkyl, alkenyl,
substituted alkenyl, cycloalkyl, substituted cycloalkyl,
cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl,
heterocyclic, and the like. Also included are amines where the two
or three substituents, together with the amino nitrogen, form a
heterocyclic or heteroaryl group.
[0045] Examples of suitable amines include, by way of example only,
isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl)
amine, tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol,
tromethamine, lysine, arginine, histidine, caffeine, procaine,
hydrabamine, choline, betaine, ethylenediamine, glucosamine,
N-alkylglucamines, theobromine, purines, piperazine, piperidine,
morpholine, N-ethylpiperidine, and the like. It should also be
understood that other carboxylic acid derivatives would be useful,
for example, carboxylic acid amides, including carboxamides, lower
alkyl carboxamides, dialkyl carboxamides, and the like.
[0046] Pharmaceutically-acceptable acid addition salts may be
prepared from inorganic and organic acids. Salts derived from
inorganic acids include hydrochloric acid, hydrobromic acid,
sulfuric acid, nitric acid, phosphoric acid, and the like. Salts
derived from organic acids include acetic acid, propionic acid,
glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid,
succinic acid, maleic acid, fumaric acid, tartaric acid, citric
acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic
acid, ethanesulfonic acid, p-toluene-sulfonic acid, salicylic acid,
and the like.
[0047] A compound may act as a pro-drug. Pro-drug means any
compound which releases an active parent drug in vivo when such
pro-drug is administered to a mammalian subject. Pro-drugs are
prepared by modifying functional groups present in such a way that
the modifications may be cleaved in vivo to release the parent
compound. Prodrugs include compounds wherein a hydroxy, amino, or
sulfhydryl group is bonded to any group that may be cleaved in vivo
to regenerate the free hydroxyl, amino, or sulfhydryl group,
respectively. Examples of prodrugs include, but are not limited to
esters (e.g., acetate, formate, and benzoate derivatives),
carbamates (e.g., N,N-dimethylamino-carbonyl) of hydroxy functional
groups, and the like.
[0048] "Treating" or "treatment" of a disease includes:
(1) preventing the disease, i.e. causing the clinical symptoms of
the disease not to develop in a mammal that may be exposed to or
predisposed to the disease but does not yet experience or display
symptoms of the disease, (2) inhibiting the disease, i.e.,
arresting or reducing the development of the disease or its
clinical symptoms, or (3) relieving the disease, i.e., causing
regression of the disease or its clinical symptoms.
[0049] A "therapeutically-effective amount" means the amount of a
compound that, when administered to a mammal for treating a
disease, is sufficient to effect such treatment for the disease.
The "therapeutically-effective amount" will vary depending on the
compound, the disease, and its severity and the age, weight, etc.,
of the mammal to be treated.
Pharmaceutical Formulations of the Compounds
[0050] In general, compounds will be administered in a
therapeutically-effective amount by any of the accepted modes of
administration for these compounds. The compounds can be
administered by a variety of routes, including, but not limited to,
oral, parenteral (e.g., subcutaneous, subdural, intravenous,
intramuscular, intrathecal, intraperitoneal, intracerebral,
intraarterial, or intralesional routes of administration), topical,
intranasal, localized (e.g., surgical application or surgical
suppository), rectal, and pulmonary (e.g., aerosols, inhalation, or
powder). Accordingly, these compounds are effective as both
injectable and oral compositions. The compounds can be administered
continuously by infusion or by bolus injection. The actual amount
of the compound, i.e., the active ingredient, will depend on a
number of factors, such as the severity of the disease, i.e., the
condition or disease to be treated, age, and relative health of the
subject, the potency of the compound used, the route and form of
administration, and other factors.
[0051] Toxicity and therapeutic efficacy of such compounds can be
determined by standard pharmaceutical procedures in cell cultures
or experimental animals, e.g., for determining the LD.sub.50 (the
dose lethal to 50% of the population) and the ED.sub.50 (the dose
therapeutically effective in 50% of the population). The dose ratio
between toxic and therapeutic effects is the therapeutic index and
it can be expressed as the ratio LD.sub.50/ED.sub.50.
[0052] The data obtained from the cell culture assays and animal
studies can be used in formulating a range of dosage for use in
humans. The dosage of such compounds lies within a range of
circulating concentrations that include the ED.sub.50 with little
or no toxicity. The dosage may vary within this range depending
upon the dosage form employed and the route of administration
utilized. For any compound used, the therapeutically-effective dose
can be estimated initially from cell culture assays. A dose may be
formulated in animal models to achieve a circulating plasma
concentration range which includes the IC.sub.50 (i.e., the
concentration of the test compound which achieves a half-maximal
inhibition of symptoms) as determined in cell culture. Such
information can be used to more accurately determine useful doses
in humans. Levels in plasma may be measured, for example, by high
performance liquid chromatography.
[0053] The amount of the pharmaceutical composition administered to
the patient will vary depending upon what is being administered,
the purpose of the administration, such as prophylaxis or therapy,
the state of the patient, the manner of administration, and the
like. In therapeutic applications, compositions are administered to
a patient already suffering from a disease in an amount sufficient
to cure or at least partially arrest the symptoms of the disease
and its complications. An amount adequate to accomplish this is
defined as "therapeutically-effective dose." Amounts effective for
this use will depend on the disease condition being treated as well
as by the judgment of the attending clinician depending upon
factors such as the severity of the inflammation, the age, weight,
and general condition of the patient, and the like.
[0054] The compositions administered to a patient are in the form
of 24 pharmaceutical compositions described supra. These
compositions may be sterilized by conventional sterilization
techniques, or may be sterile filtered. The resulting aqueous
solutions may be packaged for use as is, or lyophilized, the
lyophilized preparation being combined with a sterile aqueous
carrier prior to administration. It will be understood that use of
certain of the foregoing excipients, carriers, or stabilizers will
result in the formation of pharmaceutical salts.
[0055] The active compound is effective over a wide dosage range
and is generally administered in a pharmaceutically- or
therapeutically-effective amount. The therapeutic dosage of the
compounds will vary according to, for example, the particular use
for which the treatment is made, the manner of administration of
the compound, the health and condition of the patient, and the
judgment of the prescribing physician. For example, for intravenous
administration, the dose will typically be in the range of about
0.5 mg to about 100 mg per kilogram body weight. Effective doses
can be extrapolated from dose-response curves derived from in vitro
or animal model test systems. Typically, the clinician will
administer the compound until a dosage is reached that achieves the
desired effect.
When employed as pharmaceuticals, the compounds are usually
administered in the form of pharmaceutical compositions.
Pharmaceutical compositions contain as the active ingredient one or
more of the compounds above, associated with one or more
pharmaceutically-acceptable carriers or excipients. The excipient
employed is typically one suitable for administration to human
subjects or other mammals. In making the compositions, the active
ingredient is usually mixed with an excipient, diluted by an
excipient, or enclosed within a carrier which can be in the form of
a capsule, sachet, paper or other container. When the excipient
serves as a diluent, it can be a solid, semi-solid, or liquid
material, which acts as a vehicle, carrier, or medium for the
active ingredient. Thus, the compositions can be in the form of
tablets, pills, powders, lozenges, sachets, cachets, elixirs,
suspensions, emulsions, solutions, syrups, aerosols (as a solid or
in a liquid medium), ointments containing, for example, up to 10%
by weight of the active compound, soft and hard gelatin capsules,
suppositories, sterile injectable solutions, and sterile packaged
powders.
[0056] In preparing a formulation, it may be necessary to mill the
active compound to provide the appropriate particle size prior to
combining with the other ingredients. If the active compound is
substantially insoluble, it ordinarily is milled to a particle size
of less than 200 mesh. If the active compound is substantially
water soluble, the particle size is normally adjusted by milling to
provide a substantially uniform distribution in the formulation,
e.g., about 40 mesh. Some examples of suitable excipients include
lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum
acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium
silicate, microcrystalline cellulose, polyvinylpyrrolidone,
cellulose, sterile water, syrup, and methyl cellulose. The
formulations can additionally include: lubricating agents such as
talc, magnesium stearate, and mineral oil; wetting agents;
emulsifying and suspending agents; preserving agents such as
methyl- and propylhydroxy-benzoates; sweetening agents; and
flavoring agents. The compositions of the invention can be
formulated so as to provide quick, sustained, or delayed-release of
the active ingredient after administration to the patient by
employing procedures known in the art.
[0057] The quantity of active compound in the pharmaceutical
composition and unit dosage form thereof may be varied or adjusted
widely depending upon the particular application, the manner or
introduction, the potency of the particular compound, and the
desired concentration. The term "unit dosage forms" refers to
physically-discrete units suitable as unitary dosages for human
subjects and other mammals, each unit containing a predetermined
quantity of active material calculated to produce the desired
therapeutic effect, in association with a suitable pharmaceutical
excipient.
[0058] The compound can be formulated for parenteral administration
in a suitable inert carrier, such as a sterile physiological saline
solution. The dose administered will be determined by route of
administration.
Administration of therapeutic agents by intravenous formulation is
well known in the pharmaceutical industry. An intravenous
formulation should possess certain qualities aside from being just
a composition in which the therapeutic agent is soluble. For
example, the formulation should promote the overall stability of
the active ingredient(s), also, the manufacture of the formulation
should be cost-effective. All of these factors ultimately determine
the overall success and usefulness of an intravenous
formulation.
[0059] Other accessory additives that may be included in
pharmaceutical formulations and compounds as follow: solvents:
ethanol, glycerol, propylene glycol; stabilizers: EDTA (ethylene
diamine tetraacetic acid), citric acid; antimicrobial
preservatives: benzyl alcohol, methyl paraben, propyl paraben;
buffering agents: citric acid/sodium citrate, potassium hydrogen
tartrate, sodium hydrogen tartrate, acetic acid/sodium acetate,
maleic acid/sodium maleate, sodium hydrogen phthalate, phosphoric
acid/potassium dihydrogen phosphate, phosphoric acid/disodium
hydrogen phosphate; and tonicity modifiers: sodium chloride,
mannitol, dextrose.
[0060] The presence of a buffer is necessary to maintain the
aqueous pH in the range of from about 4 to about 8. The buffer
system is generally a mixture of a weak acid and a soluble salt
thereof, e.g., sodium citrate/citric acid; or the monocation or
dication salt of a dibasic acid, e.g., potassium hydrogen tartrate;
sodium hydrogen tartrate, phosphoric acid/potassium dihydrogen
phosphate, and phosphoric acid/disodium hydrogen phosphate.
[0061] The amount of buffer system used is dependent on (1) the
desired pH; and (2) the amount of drug.
Generally, the amount of buffer used is in a 0.5:1 to 50:1 mole
ratio of buffenalendronate (where the moles of buffer are taken as
the combined moles of the buffer ingredients, e.g., sodium citrate
and citric acid) of formulation to maintain a pH in the range of 4
to 8 and generally, a 1:1 to 10:1 mole ratio of buffer (combined)
to drug present is used. A useful buffer is sodium citrate/citric
acid in the range of 5 to 50 mg per ml. sodium citrate to 1 to 15
mg per ml. citric acid, sufficient to maintain an aqueous pH of 4-6
of the composition.
[0062] The buffer agent may also be present to prevent the
precipitation of the drug through soluble metal complex formation
with dissolved metal ions, e.g., Ca, Mg, Fe, Al, Ba, which may
leach out of glass containers or rubber stoppers or be present in
ordinary tap water. The agent may act as a competitive complexing
agent with the drug and produce a soluble metal complex leading to
the presence of undesirable particulates.
[0063] In addition, the presence of an agent, e.g., sodium chloride
in an amount of about of 1-8 mg/ml, to adjust the tonicity to the
same value of human blood may be required to avoid the swelling or
shrinkage of erythrocytes upon administration of the intravenous
formulation leading to undesirable side effects such as nausea or
diarrhea and possibly to associated blood disorders. In general,
the tonicity of the formulation matches that of human blood which
is in the range of 282 to 288 mOsm/kg, and in general is 285
mOsm/kg, which is equivalent to the osmotic pressure corresponding
to a 0.9% solution of sodium chloride.
[0064] An intravenous formulation can be administered by direct
intravenous injection, i.v. bolus, or can be administered by
infusion by addition to an appropriate `infusion solution such as
0.9% sodium chloride injection or other compatible infusion
solution.
[0065] The compositions are preferably formulated in a unit dosage
form, each dosage containing from about 5 to about 100 mg, more
usually about 10 to about 30 mg, of the active ingredient. The term
"unit dosage forms" refers to physically discrete units suitable as
unitary dosages for human subjects and other mammals, each unit
containing a predetermined quantity of active material calculated
to produce the desired therapeutic effect, in association with a
suitable pharmaceutical excipient.
[0066] The active compound is effective over a wide dosage range
and is generally administered in a pharmaceutically effective
amount. It will be understood, however, that the amount of the
compound actually administered will be determined by a physician,
in the light of the relevant circumstances, including the condition
to be treated, the chosen route of administration, the actual
compound administered, the age, weight, and response of the
individual patient, the severity of the patient's symptoms, and the
like. For preparing solid compositions such as tablets, the
principal active ingredient is mixed with a pharmaceutical
excipient to form a solid preformulation composition containing a
homogeneous mixture of a compound of the present invention. When
referring to these preformulation compositions as homogeneous, it
is meant that the active ingredient is dispersed evenly throughout
the composition so that the composition may be readily subdivided
into equally effective unit dosage forms such as tablets, pills and
capsules. This solid preformulation is then subdivided into unit
dosage forms of the type described above containing from, for
example, 0.1 to about 2000 mg of the active ingredient.
[0067] The tablets or pills may be coated or otherwise compounded
to provide a dosage form affording the advantage of prolonged
action. For example, the tablet or pill can comprise an inner
dosage and an outer dosage component, the latter being in the form
of an envelope over the former. The two components can be separated
by an enteric layer which serves to resist disintegration in the
stomach and permit the inner component to pass intact into the
duodenum or to be delayed in release. A variety of materials can be
used for such enteric layers or coatings, such materials including
a number of polymeric acids and mixtures of polymeric acids with
such materials as shellac, cetyl alcohol, and cellulose
acetate.
[0068] The liquid forms in which the novel compositions may be
incorporated for administration orally or by injection include
aqueous solutions suitably flavored syrups, aqueous or oil
suspensions, and flavored emulsions with edible oils such as
cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as
elixirs and similar pharmaceutical vehicles.
[0069] Compositions for inhalation or insufflation include
solutions and suspensions in pharmaceutically-acceptable, aqueous
or organic solvents, or mixtures thereof, and powders. The liquid
or solid compositions may contain suitable
pharmaceutically-acceptable excipients as described supra.
Compositions in pharmaceutically-acceptable solvents may be
nebulized by use of inert gases. Nebulized solutions may be
breathed directly from the nebulizing device or the nebulizing
device may be attached to a face masks tent, or intermittent
positive pressure breathing machine. Solution, suspension, or
powder compositions may be administered from devices which deliver
the formulation in an appropriate manner.
[0070] The compounds can be administered in a sustained release
form. Suitable examples of sustained-release preparations include
semipermeable matrices of solid hydrophobic polymers containing the
compounds, which matrices are in the form of shaped articles, e.g.,
films, or microcapsules. Examples of sustained-release matrices
include polyesters, hydrogels (e.g.,
poly(2-hydroxyethyl-methacrylate) as described by Langer et al., J.
Biomed. Mater. Res. 15: 167-277 (1981) and Langer, Chem. Tech. 12:
98-105 (1982) or poly(vinyl alcohol)), polylactides (U.S. Pat. No.
3,773,919), copolymers of L-glutamic acid and gamma
ethyl-L-glutamate (Sidman et al., Biopolymers 22: 547-556, 1983),
non-degradable ethylene-vinyl acetate (Langer et al., supra),
degradable lactic acid-glycolic acid copolymers such as the LUPRON
DEPOT.TM. (i.e., injectable microspheres composed of lactic
acid-glycolic acid copolymer and leuprolide acetate), and
poly-D-(-)-3-hydroxybutyric acid (EP 133,988).
[0071] The compounds can be administered in a sustained-release
form, for example a depot injection, implant preparation, or
osmotic pump, which can be formulated in such a manner as to permit
a sustained-release of the active ingredient. Implants for
sustained-release formulations are well-known in the art. Implants
may be formulated as, including but not limited to, microspheres,
slabs, with biodegradable or non-biodegradable polymers. For
example, polymers of lactic acid and/or glycolic acid form an
erodible polymer that is well-tolerated by the host.
[0072] Transdermal delivery devices ("patches") may also be
employed. Such transdermal patches may be used to provide
continuous or discontinuous infusion of the compounds in controlled
amounts. The construction and use of transdermal patches for the
delivery of pharmaceutical agents is well known in the art. See,
e.g., U.S. Pat. No. 5,023,252, issued Jun. 11, 1991, herein
incorporated by reference. Such patches may be constructed for
continuous, pulsatile, or on-demand delivery of pharmaceutical
agents.
Direct or indirect placement techniques may be used when it is
desirable or necessary to introduce the pharmaceutical composition
to the brain. Direct techniques usually involve placement of a drug
delivery catheter into the host's ventricular system to bypass the
blood-brain barrier. One such implantable delivery system used for
the transport of biological factors to specific anatomical regions
of the body is described in U.S. Pat. No. 5,011,472, which is
herein incorporated by reference. Indirect techniques usually
involve formulating the compositions to provide for drug
latentiation by the conversion of hydrophilic drugs into
lipid-soluble drugs. Latentiation is generally achieved through
blocking of the hydroxy, carbonyl, sulfate, and primary amine
groups present on the drug to render the drug more lipid-soluble
and amenable to transportation across the blood-brain barrier.
Alternatively, the delivery of hydrophilic drugs may be enhanced by
intra-arterial infusion of hypertonic solutions which can
transiently open the blood-brain barrier.
[0073] In order to enhance serum half-life, the compounds may be
encapsulated, introduced into the lumen of liposomes, prepared as a
colloid, or other conventional techniques may be employed which
provide an extended serum half-life of the compounds. A variety of
methods are available for preparing liposomes, as described in,
e.g., Szoka et al., U.S. Pat. Nos. 4,235,871, 4,501,728 and
4,837,028 each of which is incorporated herein by reference.
[0074] Pharmaceutical compositions are suitable for use in a
variety of drug delivery systems. Suitable formulations for use in
the present invention are found in Remington's Pharmaceutical
Sciences, Mace Publishing Company, Philadelphia, Pa., 17th ed.
(1985).
[0075] The provided compounds and pharmaceutical compositions show
biological activity in treating and preventing viral infections and
associated diseases, and, accordingly, have utility in treating
viral infections and associated diseases, such as Hemorrhagic fever
viruses, in mammals including humans.
[0076] Hemorrhagic fever viruses (HFVs) are RNA viruses that cause
a variety of disease syndromes with similar clinical
characteristics. HFVs that are of concern as potential biological
weapons include but are not limited to: Arenaviridae (Junin,
Machupo, Guanarito, Sabia, and Lassa), Filoviridae (Ebola and
Marburg viruses), Flaviviridae (yellow fever, Omsk hemorrhagic
fever and Kyasanur Forest disease viruses), and Bunyaviridae (Rift
Valley fever and Crimean-Congo hemorrhagic fever). The
naturally-occurring arenaviruses and potential engineered
arenaviruses are included in the Category A Pathogen list according
to the Centers for Disease Control and Prevention as being among
those agents that have greatest potential for mass casualties.
[0077] Risk factors include: travel to Africa or Asia, handling of
animal carcasses, contact with infected animals or people, and/or
arthropod bites. Arenaviruses are highly infectious after direct
contact with infected blood and/or bodily secretions. Humans
usually become infected through contact with infected rodents, the
bite of an infected arthropod, direct contact with animal
carcasses, inhalation of infectious rodent excreta and/or injection
of food contaminated with rodent excreta. The Tacaribe virus has
been associated with bats. Airborne transmission of hemorrhagic
fever is another mode. Person-to-person contact may also occur in
some cases.
[0078] All of the hemorrhagic fevers exhibit similar clinical
symptoms. However, in general the clinical manifestations are
non-specific and variable. The incubation period is approximately
7-14 days. The onset is gradual with fever and malaise, tachypnea,
relative bradycardia, hypotension, circulatory shock, conjunctival
infection, pharyngitis, lymphadenopathy, encephalitis, myalgia,
back pain, headache and dizziness, as well as hyperesthesia of the
skin. Some infected patients may not develop hemorrhagic
manifestations.
[0079] Methods of diagnosis at specialized laboratories include
antigen detection by antigen-capture enzyme-linked immunosorbent
assay (ELISA), IgM antibody detection by antibody-capture
enzyme-linked immunosorbent assay, reverse transcriptase polymerase
chain reaction (RT-PCR), and viral isolation. Antigen detection (by
enzyme-linked immunosorbent assay) and reverse transcriptase
polymerase chain reaction are the most useful diagnostic techniques
in the acute clinical setting. Viral isolation is of limited value
because it requires a biosafety level 4 (BSL-4) laboratory.
Example 1
Determining Anti Dengue-2 Activity of Compounds of the
Invention
[0080] A sensitive and reproducible high-throughput screening (HTS)
assay has been established to measure dengue virus-induced
cytopathic effect (CPE). To determine the amount of dengue virus
stock required to produce complete CPE in 5 days, Vero cell
monolayers were seeded on 96-well plates and infected with 10-fold
serial dilutions of the dengue virus stock representing a
multiplicity of infection (MOI) of approximately 0.001 PFU/cell to
0.1 PFU/cell. At 5 days post-infection, the cultures were fixed
with 5% glutaraldehyde and stained with 0.1% crystal violet.
Virus-induced CPE was quantified spectrophometrically at
OD.sub.570. From this analysis, an MOI of 0.1 PFU/cell of dengue
virus stock was chosen for use in the HTS assay. To establish the
signal-to-noise ratio (S/N) of the 96-well assay and evaluate the
well-to-well and assay-to-assay variability, five independent
experiments were performed. Vero cell monolayers were infected with
0.1 PFU/cell of dengue virus stock. Each plate contained the
following controls: quadruplicate virus-infected wells,
quadruplicate uninfected cell wells and a dose response curve in
duplicate for ribavirin at 500, 250, 125 and 62 .mu.M, as reference
standards. At day 5 post-infection, the plates were processed as
described above.
[0081] Compounds were dissolved in DMSO and diluted in medium such
that the final concentration in each well was 5 .mu.M compound and
0.5% DMSO. The compounds were added robotically to the culture
medium using the PerkinElmer MultiPROBE.RTM. II HT PLUS robotic
system. Following compound addition, cultures were infected with
dengue virus (DEN-2 strain New Guinea C). After 5 days incubation,
plates were processed and CPE quantified on a PerkinElmer EnVision
II plate reader system.
[0082] The results of these experiments indicated that the 96-well
assay format is robust and reproducible. The S/N ratio (ratio of
signal of cell control wells (signal) to virus control wells
(noise)) was 5.0.+-.1.2. The well-to-well variability was
determined for each individual plate and found to have a
coefficient of variance of less than 10% for both positive control
and negative control wells, and overall assay-to-assay variability
was less than 15%. Using this assay, the EC.sub.50 values for
ribavirin were determined to be 125.+-.25 .mu.M, respectively. The
effectiveness of ribavirin against dengue varies with the cell type
used, but the values obtained were within the range of published
values for this compound (2, 14, 32). Taken together, these results
show that a sensitive and reproducible HTS assay has been
successfully developed to evaluate our compound library for
inhibitors of dengue virus replication.
[0083] This assay was the basis of a high-throughput screen for
dengue virus inhibitors, against which a library of 210,000
compounds was tested. Compounds that inhibited dengue virus induced
CPE by at least 50% were further investigated for chemical
tractability, potency, and selectivity.
[0084] Initially, the chemical structures of the hit compounds were
examined for chemical tractability. A chemically tractable compound
is defined as one that is synthetically accessible using reasonable
chemical methodology, and which possesses chemically stable
functionalities and potential drug-like qualities. Hits that passed
this medicinal chemistry filter were evaluated for their potency.
Compound potency was determined by evaluating inhibitory activity
across a broad range of concentrations. Nonlinear regression was
used to generate best-fit inhibition curves and to calculate the
50% effective concentration (EC50).
[0085] Compounds that were active in the primary screen were also
tested for activity in viral yield assays. Table 1 below shows some
of the compounds that were tested for activity against Dengue-2
(Strain New Guinea C) in a viral yield assay at a range of
concentrations. Vero cells in 12-well plates were infected with
dengue-2 virus at a multiplicity of infection (MOI) of 0.1, treated
with compound (or DMSO as a control), incubated at 37.degree. C.,
harvested 48 hours post infection and titered on Vero cells as
described above. The EC50 was calculated through ExcelFit. Those
compounds with activity below 1 .mu.M are indicated with "A", those
with activity between 1 and 10 .mu.M are indicated with "B", those
with activity between 10 and 25 .mu.M with "C", and those with
activity above 25 .mu.M are indicated with "D".
TABLE-US-00001 TABLE 1 List of compounds of the present invention
and their anti-dengue 2 viral activity. Activity A: EC.sub.50 <
1 uM; B: 1 < EC.sub.50 < 10 uM; Mole- C: 10 < EC.sub.50
< Com- cular 25 uM; pound Chemical Structure Weight Chemical
Name D: EC.sub.50 > uM 1 ##STR00004## 414.5
N-(4-Diethylsulfamoyl- phenyl)-2-(4-oxo-4H- quinazolin-3-yl)-
acetamide A 2 ##STR00005## 328.4 N-(5-Diethylsulfamoyl-
2-methoxy-phenyl)- isobutyramide A 3 ##STR00006## 415.5
N-(4-Diethylsulfamoyl- phenyl)-2-(4-oxo-4H- benzo[d][1,2,3]triazin-
3-yl)-acetamide B 4 ##STR00007## 448.5 N-(4-Diethylsulfamoyl-
phenyl)-2-(1,3-dimethyl- 2,6-dioxo-1,2,3,6- tetrahydro-purin-7-yl)-
acetamide B 5 ##STR00008## 420.5 N-(4-Diethylsulfamoyl-
phenyl)-2-(4-oxo-4H- thieno[2,3-d]pyrimidin- 3-yl)-acetamide B 6
##STR00009## 448.9 2-(6-Chloro-4-oxo-4H- quinazolin-3-yl)-N-[4-
(isopropyl-methyl- sulfamoyl)-phenyl]- acetamide B 7 ##STR00010##
432.5 2-(7-Fluoro-4-oxo-4H- quinazolin-3-yl)-N-[4-
(isopropyl-methyl- sulfamoyl)-phenyl]- acetamide B 8 ##STR00011##
430.5 N-(4-Diethylsulfamoyl- phenyl)-2-(1,4-dioxo- 3,4-dihydro-1H-
phthalazin-2-yl)- acetamide B 9 ##STR00012## 465.5
N-(4-Diethylsulfamoyl- phenyl)-2-(1,3-dioxo- 1H,3H-benzo[de]
isoquinolin-2-yl)- acetamide B 10 ##STR00013## 448.6
N-(4-Diethylsulfamoyl- phenyl)-2-(5,6-dimethyl-
4-oxo-4H-thieno[2,3-d] pyrimidin-3-yl)- acetamide B 11 ##STR00014##
448.6 N-(4-Diethylsulfamoyl- phenyl)-2-(6-ethyl-4-
oxo-4H-thieno[2,3- d]pyrimidin-3-yl)- acetamide B 12 ##STR00015##
473.5 N-(5-Diethylsulfamoyl- 2-methyl-phenyl)-2-(7- nitro-4-oxo-4H-
quinazolin-3-yl)- acetamide B 13 ##STR00016## 478.5
N-(5-Diethylsulfamoyl- 2-methoxy-phenyl)-2- (1,3-dimethyl-2,6-
dioxo-1,2,3,6-tetrahydro- purin-7-yl)-acetamide B 14 ##STR00017##
328.4 N-(5-Diethylsulfamoyl- 2-methoxy-phenyl)- butyramide B 15
##STR00018## 342.5 N-(5-Diethylsulfamoyl- 2-methoxy-phenyl)-3-
methyl-butyramide B 16 ##STR00019## 376.5 N-(5-Diethylsulfamoyl-
2-methoxy-phenyl)-2- phenyl-acetamide B 17 ##STR00020## 440.5
1-[2-(4-Oxo-4H- quinazolin-3-yl)- acetyl]-2,3-dihydro-
1H-indole-5-sulfonic acid diethylamide B 18 ##STR00021## 454.5
N-[4-(2,6-Dimethyl- piperidine-1- sulfonyl)-phenyl]-2-
(4-oxo-4H-quinazolin- 3-yl)-acetamide B 19 ##STR00022## 454.5
N-[4-(3,5-Dimethyl- piperidine-1- sulfonyl)-phenyl]-2-
(4-oxo-4H-quinazolin- 3-yl)-acetamide B 20 ##STR00023## 445.5
N-(5-Diethylsulfamoyl- 2-methoxy-phenyl)-2- (4-oxo-4H-benzo[d]
[1,2,3]triazin-3-yl)- acetamide B 21 ##STR00024## 342.5 Pentanoic
acid (5- diethylsulfamoyl-2- methoxy-phenyl)- amide C 22
##STR00025## 420.5 N-(4-Diethylsulfamoyl- phenyl)-2-(4-oxo-4H-
thieno[3,2-d]pyrimidin- 3-yl)-acetamide C 23 ##STR00026## 448.9
2-(7-Chloro-4-oxo- 4H-quinazolin-3-yl)- N-(4-diethylsulfamoyl-
phenyl)-acetamide C 24 ##STR00027## 454.5 N-(4-Diethylsulfamoyl-
phenyl)-2-(4-oxo-4H- benzo[4,5]furo[3,2-d] pyrimidin-3-yl)-
acetamide C 25 ##STR00028## 487.6 N-(5-Diethylsulfamoyl-
2-isopropoxy-phenyl)- 3-(4-oxo-4H-benzo[d] [1,2,3]triazin-3-yl)-
propionamide C 26 ##STR00029## 428.5 N-(4-Diethylsulfamoyl-
phenyl)-2-(8-methyl-4- oxo-4H-quinazolin-3- yl)-acetamide C 27
##STR00030## 459.5 N-(5-Diethylsulfamoyl- 2-methoxy-phenyl)-3-
(4-oxo-4H-benzo[d] [1,2,3]triazin-3-yl)- propionamide C 28
##STR00031## 460.6 N-[4-(2,6-Dimethyl- piperidine-1-sulfonyl)-
phenyl]-2-(4-oxo-4H- thieno[2,3-d]pyrimidin- 3-yl)-acetamide C 29
##STR00032## 471.6 N-(4-Diethylsulfamoyl- phenyl)-2-[ethyl-(4-
oxo-3,4-dihydro- quinazolin-2-ylmethyl)- amino]-acetamide C 30
##STR00033## 440.4 2-(4-Oxo-4H- quinazolin-3-yl)-N-[4-
(2,2,2-trifluoro-ethyl- sulfamoyl)-phenyl]- acetamide C 31
##STR00034## 412.5 2-(4-Oxo-4H- quinazolin-3-yl)-N-(4-
(pyrrolidine-1- sulfonyl)-phenyl]- acetamide D 32 ##STR00035##
441.5 2-(4-Oxo-4H- quinazolin-3-yl)-N-[4- (thiazol-2-ylsulfamoyl)-
phenyl]-acetamide D 33 ##STR00036## 464.5 N-[4-(4,6-Dimethyl-
pyrimidin-2-yl- sulfamoyl)-phenyl]- 2-(4-oxo-4H- quinazolin-3-yl)-
acetamide D 34 ##STR00037## 428.5 N-[4-(Morpholine-4-
sulfonyl)-phenyl]-2- (4-oxo-4H-quinazolin- 3-yl)-acetamide D 35
##STR00038## 428.5 N-[4-(Butyl-methyl- sulfamoyl)-phenyl]-
2-(4-oxo-4H- quinazolin-3-yl)- acetamide D 36 ##STR00039## 462.5
N-[4-(Benzyl-methyl- sulfamoyl)-phenyl]-2- (4-oxo-4H-quinazolin-
3-yl)-acetamide D 37 ##STR00040## 440.5 N-[4-(2-Methyl-
piperidine-1-sulfonyl)- phenyl]-2-(4-oxo-4H- quinazolin-3-yl)-
acetamide D 38 ##STR00041## 358.4 2-(4-Oxo-4H- quinazolin-3-yl)-N-
(4-sulfamoyl-phenyl)- acetamide D 39 ##STR00042## 436.5
2-(4-Oxo-4H- quinazolin-3-yl)-N- [4-(pyrimidin-2- ylsulfamoyl)-
phenyl]-acetamide D 40 ##STR00043## 485.6 N-[4-(3-Morpholin-
4-yl-propylsulfamoyl)- phenyl]-2-(4-oxo-4H- quinazolin-3-yl)-
acetamide D 41 ##STR00044## 438.5 N-{4-[(Furan-2-
ylmethyl)-sulfamoyl]- phenyl}-2-(4-oxo-4H- quinazolin-3-yl)-
acetamide D 42 ##STR00045## 414.5 N-[4-(Isopropyl-methyl-
sulfamoyl)-phenyl]-2- (4-oxo-4H-quinazolin- 3-yl)-acetamide D 43
##STR00046## 480.6 N-[4-(1-Bicyclo[2.2.1] hept-2-yl-ethyl-
sulfamoyl)-phenyl]-2- (4-oxo-4H-quinazolin- 3-yl)-acetamide D 44
##STR00047## 400.5 N-(4-Isopropyl- sulfamoyl-phenyl)- 2-(4-oxo-4H-
quinazolin-3-yl)- acetamide D 45 ##STR00048## 441.5 N-[4-(4-Methyl-
piperazine-1-sulfonyl)- phenyl]-2-(4-oxo-4H- quinazolin-3-yl)-
acetamide D 46 ##STR00049## 372.4 N-(4-Methylsulfamoyl-
phenyl)-2-(4-oxo-4H- quinazolin-3-yl)- acetamide D 47 ##STR00050##
440.5 N-[4-(4-Methyl- piperidine-1- sulfonyl)-phenyl]- 2-(4-oxo-4H-
quinazolin-3-yl)- acetamide D 48 ##STR00051## 476.5
N-[4-(1,1-Dioxo- tetrahydro- 1lambda*6*- thiophen-3-yl-
sulfamoyl)-phenyl]- 2-(4-oxo-4H- quinazolin-3-yl)- acetamide D 49
##STR00052## 454.5 N-[4-(Cyclohexyl- methyl-sulfamoyl)-
phenyl]-2-(4-oxo- 4H-quinazolin-3- yl)-acetamide D 50 ##STR00053##
386.4 N-(4-Dimethyl- sulfamoyl-phenyl)- 2-(4-oxo-4H-
quinazolin-3-yl)- acetamide D 51 ##STR00054## 440.5
N-[4-(Azepane-1- sulfonyl)-phenyl]- 2-(4-oxo-4H- quinazolin-3-yl)-
acetamide D 52 ##STR00055## 426.5 2-(4-Oxo-4H- quinazolin-3-yl)-N-
[4-(piperidine-1- sulfonyl)-phenyl]- acetamide D 53 ##STR00056##
440.5 N-[4-(3-Methyl- piperidine-1- sulfonyl)-phenyl]- 2-(4-oxo-4H-
quinazolin-3-yl)- acetamide D 54 ##STR00057## 430.4
{4-[2-(4-Oxo-4H- quinazolin-3-yl)- acetylamino]- benzenesulfonyl-
amino}-acetic acid methyl ester D 55 ##STR00058## 414.5
N-(3-tert-Butyl- sulfamoyl-phenyl)- 2-(4-oxo-4H- quinazolin-3-yl)-
acetamide D 56 ##STR00059## 496.6 N-(4-Diethylsulfamoyl-
phenyl)-2-(4-oxo-6- phenyl-4H-thieno[2,3- d]pyrimidin-3-yl)-
acetamide D 57 ##STR00060## 432.5 N-(4-Diethylsulfamoyl-
phenyl)-2-(7-fluoro-4- oxo-4H-quinazolin-3- yl)-acetamide D 58
##STR00061## 474.5 N-(4-Diethylsulfamoyl- phenyl)-2-(6,7-
dimethoxy-4-oxo-4H- quinazolin-3-yl)- acetamide D 59 ##STR00062##
496.6 N-(4-Diethylsulfamoyl- phenyl)-2-(4-oxo-5-
phenyl-4H-thieno[2,3- d]pyrimidin-3-yl)- acetamide D 60
##STR00063## 488.5 N-(4-Diethylsulfamoyl- phenyl)-2-(7-methyl-
amino-6-nitro-4-oxo- 4H-quinazolin-3-yl)- acetamide D 61
##STR00064## 526.6 N-(4-Diethylsulfamoyl- phenyl)-2-[6-(4-
methoxy-phenyl)-4- oxo-4H-thieno[3,2-d] pyrimidin-3-yl]- acetamide
D 62 ##STR00065## 400.5 N-(4-Dimethylsulfamoyl-
phenyl)-2-(8-methyl-4- oxo-4H-quinazolin- 3- yl)-acetamide D 63
##STR00066## 417.5 N-[(4-Diethylsulfamoyl- phenylcarbamoyl)-
methyl]-3,5- dimethyl-benzamide D 64 ##STR00067## 474.6
N-(4-Diethylsulfamoyl- phenyl)-2-(3-ethyl-4- oxo-3,4-dihydro-
quinazolin-2-yl- sulfanyl)-acetamide D 65 ##STR00068## 414.5
N-(3-Diethylsulfamoyl- phenyl)-2-(4-oxo-4H- quinazolin-3-yl)-
acetamide D 66 ##STR00069## 472.5 4-Diethylsulfamoyl- benzoic acid
3- carbamoylmethyl-4- oxo-3,4-dihydro- quinazolin-2-ylmethyl ester
D 67 ##STR00070## 400.5 4-Diethylsulfamoyl-N- (4-oxo-4H-
quinazolin-3-yl)- benzamide D 68 ##STR00071## 457.6
N-[1-(4-Diethyl- sulfamoyl-phenyl)- ethyl]-3-(4-oxo-4H-
benzo[d][1,2,3] triazin-3-yl)- propionamide D 69 ##STR00072## 428.5
N-(4-Diethylsulfamoyl- benzyl)-2-(4-oxo-4H- quinazolin-3-yl)-
acetamide D 70 ##STR00073## 496.6 N-[(4-Diethylsulfamoyl-
phenylcarbamoyl)- methyl]-3- dimethylsulfamoyl- benzamide D 71
##STR00074## 460.6 N-(4-Diethylsulfamoyl- phenyl)-2-(3-methyl-
4-oxo-3,4-dihydro- quinazolin-2-yl- sulfanyl)-acetamide D 72
##STR00075## 403.5 N-[(4-Diethylsulfamoyl- phenylcarbamoyl)-
methyl]-4-methyl- benzamide D 73 ##STR00076## 510.6
N-[(4-Diethylsulfamoyl- phenylcarbamoyl)- methyl]-4-isopropyl-
sulfamoyl-benzamide D 74 ##STR00077## 439.5
Naphthalene-2-carboxylic acid [(4-diethylsulfamoyl-
phenylcarbamoyl)- methyl]-amide D 75 ##STR00078## 506.0
2-[(7-Chloro-4-oxo-3,4- dihydro-quinazolin-2-
ylmethyl)-ethyl-amino]- N-(4-diethylsulfamoyl- phenyl)-acetamide D
76 ##STR00079## 460.6 4-{[(4-Diethylsulfamoyl- phenylcarbamoyl)-
methyl]-amino}-N,N- diethyl-benzamide D 77 ##STR00080## 429.5
N-(4-Diethylsulfamoyl- phenyl)-3-(4-oxo-4H- benzo[d][1,2,3]triazin-
3-yl)-propionamide D 78 ##STR00081## 458.5 N-(4-Diethylsulfamoyl-
phenyl)-2-(3-ethyl-2,4- dioxo-3,4-dihydro-2H- quinazolin-1-yl)-
acetamide D 79 ##STR00082## 449.5 N-[(4-Diethylsulfamoyl-
phenylcarbamoyl)- methyl]-3,4- dimethoxy-benzamide D
80 ##STR00083## 465.6 Biphenyl-4-carboxylic acid [(4-diethyl-
sulfamoyl-phenyl- carbamoyl)-methyl]- amide D 81 ##STR00084## 473.5
(4-Oxo-4H-benzo[d] [1,2,3]triazin-3-yl)- acetic acid (4-
diethylsulfamoyl- phenylcarbamoyl)- methyl ester D 82 ##STR00085##
429.5 N-[4-(Morpholine-4- sulfonyl)-phenyl]-2- (4-oxo-4H-benzo[d]
[1,2,3]triazin-3-yl)- acetamide D 83 ##STR00086## 481.6
N-[4-(1-Bicyclo[2.2.1] hept-2-yl-ethyl- sulfamoyl)-phenyl]-2-
(4-oxo-4H-benzo[d] [1,2,3]triazin-3-yl)- acetamide D 84
##STR00087## 403.4 N-[4-(Methoxy-methyl- sulfamoyl)-phenyl]-2-
(4-oxo-4H-benzo[d] [1,2,3]triazin-3-yl)- acetamide D 85
##STR00088## 387.4 N-(4-Dimethylsulfamoyl- phenyl)-2-(4-oxo-4H-
benzo[d][1,2,3]triazin- 3-yl)-acetamide D 86 ##STR00089## 415.5
N-[4-(Isopropyl-methyl- sulfamoyl)-phenyl]-2-(4- oxo-4H-benzo[d]
[1,2,3]triazin-3-yl)- acetamide D 87 ##STR00090## 427.5
2-(4-Oxo-4H- benzo[d][1,2,3]triazin- 3-yl)-N-[4-(piperidine-
1-sulfonyl)-phenyl]- acetamide D 88 ##STR00091## 457.6
N-(5-Diethylsulfamoyl- 2-dimethylamino- phenyl)-2-(4-oxo-
4H-quinazolin-3-yl)- acetamide D 89 ##STR00092## 428.5
N-(5-Diethylsulfamoyl- 2-methyl-phenyl)-2- (4-oxo-4H-quinazolin-
3-yl)-acetamide D 90 ##STR00093## 442.5 N-(5-Diethylsulfamoyl-
2,3-dimethyl-phenyl)- 2-(4-oxo-4H- quinazolin-3-yl)- acetamide D 91
##STR00094## 444.5 N-(5-Diethylsulfamoyl- 2-methoxy-phenyl)-2-
(4-oxo-4H- quinazolin-3-yl)- acetamide D 92 ##STR00095## 458.5
N-(5-Diethylsulfamoyl- 2-ethoxy-phenyl)-2- (4-oxo-4H-quinazolin-
3-yl)-acetamide D 93 ##STR00096## 448.9 N-(2-Chloro-5-
diethylsulfamoyl- phenyl)-2-(4-oxo-4H- quinazolin-3-yl)- acetamide
D 94 ##STR00097## 431.5 N-(5-Diethylsulfamoyl- 2-hydroxy-phenyl)-2-
(4-oxo-4H-benzo[d] [1,2,3]triazin-3-yl)- acetamide D 95
##STR00098## 429.5 N-(5-Diethylsulfamoyl- 2-methyl-phenyl)-2-
(4-oxo-4H-benzo[d] [1,2,3]triazin-3-yl)- acetamide D 96
##STR00099## 426.5 1-(3-Methyl-4-oxo- 3,4-dihydro- quinazolin-2-yl-
methyl)-2,3- dihydro-1H-indole- 5-sulfonic acid diethylamide D 97
##STR00100## 468.6 1-[2-(8-Methyl-4- oxo-4H-quinazolin-
3-yl)-acetyl]-1,2,3,4- tetrahydro-quinoline- 6-sulfonic acid
diethylamide D 98 ##STR00101## 476.6 N-[4-(Benzyl-methyl-
sulfamoyl)-phenyl]-3- (4-oxo-4H-quinazolin- 3-yl)-propionamide D 99
##STR00102## 405.5 N-(3-Benzyl-4-oxo- 3,4-dihydro-
quinazolin-6-yl)- 4-methyl- benzenesulfonamide D 100 ##STR00103##
372.4 2-(4-Methyl-1-oxo- 1H-phthalazin- 2-yl)-N-(4-sulfamoyl-
phenyl)-acetamide D 101 ##STR00104## 430.5 2-(3-Ethyl-2,4-dioxo-
3,4-dihydro-2H- quinazolin-1-yl)-N- [2-(4-sulfamoyl-
phenyl)-ethyl]- acetamide D 102 ##STR00105## 416.5
2-(3-Methyl-2,4-dioxo- 3,4-dihydro-2H- quinazolin-1-yl)-N-[2-
(4-sulfamoyl-phenyl)- ethyl]-acetamide D 103 ##STR00106## 400.5
N-{2-[4-(3-Methyl- 4-oxo-3,4-dihydro- quinazolin-6-
ylsulfamoyl)-phenyl]- ethyl}-acetamide D 104 ##STR00107## 476.6
N-{2-[4-(3-Benzyl- 4-oxo-3,4-dihydro- quinazolin-6- ylsulfamoyl)-
phenyl]-ethyl}- acetamide D 105 ##STR00108## 428.5
N-(3-Diethylsulfamoyl- 4-methyl-phenyl)-2- (4-oxo-4H-quinazolin-
3-yl)-acetamide D 106 ##STR00109## 358.4 2-(4-Oxo-4H-quinazolin-
3-yl)-N-(3-sulfamoyl- phenyl)-acetamide D 107 ##STR00110## 386.4
N-(3-Dimethylsulfamoyl- phenyl)-2-(4-oxo-4H- quinazolin-3-yl)-
acetamide D 108 ##STR00111## 402.4 4-Oxo-3-[(4-sulfamoyl-
phenylcarbamoyl)- methyl]-3,4-dihydro- phthalazine-1- carboxylic
acid D 109 ##STR00112## 372.4 3-(4-Oxo-3,4-dihydro-
quinazolin-2-yl)-N-(4- sulfamoyl-phenyl)- propionamide D 110
##STR00113## 403.4 2-(7-Nitro-4-oxo-4H- quinazolin-3-yl)-N-
(4-sulfamoyl-phenyl)- acetamide D 111 ##STR00114## 416.4
(4-Oxo-4H-quinazolin- 3-yl)-acetic acid (4- sulfamoyl-phenyl-
carbamoyl)-methyl ester D 112 ##STR00115## 400.5
4-Dimethylsulfamoyl- N-methyl-N-(4-oxo- 3,4-dihydro-quinazolin-
2-ylmethyl)-benzamide D 113 ##STR00116## 413.5 2-(4-Oxo-4H-
benzo[d][1,2,3]triazin- 3-yl)-N-[4-(pyrrolidine-
1-sulfonyl)-phenyl]- acetamide D 114 ##STR00117## 443.5
2-[Ethyl-(4-oxo-3,4- dihydro-quinazolin- 2-ylmethyl)-amino]-
N-[2-(4-sulfamoyl- phenyl)-ethyl]- acetamide D 115 ##STR00118##
443.5 N-(4-Dimethyl- sulfamoyl-phenyl)- 2-[ethyl-(4-oxo-
3,4-dihydro- quinazolin-2-ylmethyl)- amino]-acetamide D 116
##STR00119## 430.4 3-(4-Oxo-3,4-dihydro- quinazolin-2-yl)-
propionic acid (4- sulfamoyl- phenylcarbamoyl)- methyl ester D 117
##STR00120## 477.5 (4-Oxo-4H-quinazolin- 3-yl)-acetic acid 4-
[(3,4-dimethyl- benzenesulfonyl)- methyl-amino]- phenyl ester D 118
##STR00121## 503.6 2-[(6,7-Dimethoxy-4- oxo-3,4-dihydro-
quinazolin-2-ylmethyl)- ethyl-amino]-N-[2-(4- sulfamoyl-phenyl)-
ethyl]-acetamide D 119 ##STR00122## 358.4 2-(2-Oxo-2H-
quinoxalin-1-yl)-N- (4-sulfamoyl- phenyl)-acetamide D 120
##STR00123## 475.5 2-[(6,7-Dimethoxy- 4-oxo-3,4-dihydro-
quinazolin-2-ylmethyl)- ethyl-amino]-N-(4- sulfamoyl-phenyl)-
acetamide D 121 ##STR00124## 412.5 2-(4-Oxo-4H- quinazolin-3-yl)-N-
[3-(pyrrolidine-1- sulfonyl)-phenyl]- acetamide D 122 ##STR00125##
448.5 N-[3-(Methyl-phenyl- sulfamoyl)-phenyl]- 2-(4-oxo-4H-
quinazolin-3-yl)- acetamide D 123 ##STR00126## 457.5
2-(7-Nitro-4-oxo-4H- quinazolin-3-yl)-N- [4-(pyrrolidine-1-
sulfonyl)-phenyl]- acetamide D 124 ##STR00127## 470.5
(4-Oxo-4H-quinazolin- 3-yl)-acetic acid [4- (pyrrolidine-1-
sulfonyl)-phenyl- carbamoyl]- methyl ester D 125 ##STR00128## 474.5
(4-Oxo-4H-quinazolin- 3-yl)-acetic acid (5- dimethylsulfamoyl-
2-methoxy-phenyl carbamoyl)-methyl ester D 126 ##STR00129## 461.5
N-(5-Dimethyl- sulfamoyl-2- methoxy-phenyl)-2- (7-nitro-4-oxo-4H-
quinazolin-3-yl)- acetamide D 127 ##STR00130## 359.4
4-[2-(4-Oxo-4H- quinazolin-3-yl)- acetylamino]- benzenesulfonic
acid D 128 ##STR00131## 401.4 (4-Oxo-4H-quinazolin- 3-yl)-acetic
acid 3- dimethylsulfamoyl- benzyl ester D 129 ##STR00132## 434.9
2-(6-Chloro-4-oxo-4H- quinazolin-3-yl)-N- (5-dimethylsulfamoyl-
2-methyl-phenyl)- acetamide D 130 ##STR00133## 386.4 2-(2-Oxo-2H-
quinoxalin-1-yl)-N- [2-(4-sulfamoyl- phenyl)-ethyl]- acetamide D
131 ##STR00134## 359.4 2-(4-Oxo-4H- benzo[d][1,2,3]triazin-
3-yl)-N-(3-sulfamoyl- phenyl)-acetamide D 132 ##STR00135## 414.5
N-Methy1-3-(4-oxo- 3,4-dihydro- quinazolin-2-yl)-N-
[1-(4-sulfamoyl- phenyl)-ethyl]- propionamide D 133 ##STR00136##
457.6 N-(4-Isopropyl- sulfamoyl-phenyl)- 2-[methyl-(4-oxo-
3,4-dihydro- quinazolin-2-yl- methyl)-amino]- propionamide D 134
##STR00137## 471.5 2-(6-Nitro-4-oxo-4H- quinazolin-3-yl)-N-
[4-(piperidine-1- sulfonyl)-phenyl]- acetamide D 135 ##STR00138##
473.5 N-[4-(Morpholine-4- sulfonyl)-phenyl]-2- (6-nitro-4-oxo-4H-
quinazolin-3-yl)- acetamide D 136 ##STR00139## 388.4
N-(4-Methoxy-3- sulfamoyl-phenyl)-2- (4-oxo-4H-quinazolin-
3-yl)-acetamide D 137 ##STR00140## 476.6 3-(4-Oxo-3-p-tolyl-
3,4-dihydro- quinazolin-2-yl)-N- (4-sulfamoyl- benzyl)-propionamide
D 138 ##STR00141## 475.0 N-[4-(Azepane-1- sulfonyl)-phenyl]-
2-(6-chloro-4-oxo- 4H-quinazolin-3-yl)- acetamide D 139
##STR00142## 458.5 N-[4-(Azepane-1- sulfonyl)-phenyl]-
2-(7-fluoro-4-oxo- 4H-quinazolin-3-yl)- acetamide D
Example 2
Determining Selectivity or Specificity of Compounds of the Present
Invention
[0086] Those compounds with activity against dengue-2 at effective
concentrations of less than 10 .mu.M as identified in Example 1
above were tested for activity against each serotype of dengue in a
viral yield assay to generate EC50 values (Table 2). Select
compounds were also tested for more broad spectrum activity against
other members of the Flaviviridae family including Modoc, which is
a murine flavivirus, as well as Bovine Viral Diarrhea Virus (BVDV),
which is a Pestivirus. Since dengue virus is able to replicate in
multiple cell lines and to ensure that the activity seen in vero
cells is consistent, select compounds were also tested for their
effective concentration in a viral yield assay against dengue-2 in
C6/36 mosquito cells.
TABLE-US-00002 TABLE 2 Select Compounds activities against Den-1,
Den-2, Den-3, Den-4, Modoc and BVDV. EC50/90 Com- (.mu.M)
Specificity pound MW Structure Yield (.mu.M) 1 414.48 MW
##STR00143## 4.3/10.7 Den-1 0.3/5.2 Den-2 1.0/8.2 Den-3 1.7/4.1
Den-4 1.2 Modoc >50 BVDV 0.3 C6/36 2 328.43 MW ##STR00144##
7.6/8.9 Den-1 0.3/4.1 Den-2 5.5/13.3 Den-3 2.1/12.5 Den-4 6.2 Modoc
>50 BVDV 0.7 C6/36 3 415.47 MW ##STR00145## 1.3/5.1 Den-1
4.0/10.0 Den-2 1.0/12.1 Den-3 2.3/10.0 Den-4 3.1 Modoc >50 BVDV
4 448.50 MW ##STR00146## 7.8/8.9 Den-2 13.5/22.6 Den-3 4.3/22.7
Den-4 >25 Modoc >50 BVDV 6 448.93 MW ##STR00147## 1.8/43.2
Den-2 64 BVDV 7 432.42 MW ##STR00148## 4.6/>50 Den-2 >50 BVDV
8 430.48 MW ##STR00149## 2.8/25.6 Den-2 >50 BVDV 9 465.52 MW
##STR00150## 1.3/7.3 Den-2 >50 BVDV 10 448.5 MW ##STR00151##
1.25/>50 Den-2 11 446.5 MW ##STR00152## 4.9/>50 Den-2 12
473.51 MW ##STR00153## >25/>25 Den-1 11.1/>25 Den-2
2.9/>25 Den-3 5.2/14.6 Den-4 >25 Modoc >50 BVDV 13 478.53
MW ##STR00154## 7.1/8.5 Den-1 2.4/10.3 Den-2 4.1/9.4 Den-3 1.9/9.2
Den-4 >25 Modoc 47.7 BVDV 2.1 C6/36 14 328.43 MW ##STR00155##
>25/>25 Den-1 3.6/27.5 Den-2 4.2/14.9 Den-3 4.8/11.1 Den-4
>25 Modoc >50 BVDV 15 342.46 MW ##STR00156## 2.4/>100
Den-2 1.2/37.2 Den-4 10.1 Modoc >50 BVDV 16 376.47 MW
##STR00157## Den-1 5.8/10.3 Den-2 Den-3 2.1/6.4 Den-4 3.6 Modoc
>50 BVDV 17 ##STR00158## Den-1 5.1/12.1 Den-2 Den-3 Den-4 20
449.94 MW ##STR00159## Den-1 1.3/20 Den-2 Den-3 Den-4 21 342.46 MW
##STR00160## Den-1 14.2/24.1 Den-2 Den-3 8.2/12.0 Den-4 6 Modoc
>50 BVDV
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[0122] All references cited herein are herein incorporated by
reference in their entirety for all purposes.
[0123] The invention has been described in terms of preferred
embodiments thereof, but is more broadly applicable as will be
understood by those skilled in the art. The scope of the invention
is only limited by the following claims.
* * * * *
References