U.S. patent application number 13/988707 was filed with the patent office on 2013-12-26 for methods of treating cancer and other disorders.
This patent application is currently assigned to Waake Forest University Health Sciences. The applicant listed for this patent is Steven J. Kridel, W. Todd Lowther, Herman W. Odens, Jeffrey D. Schmitt. Invention is credited to Steven J. Kridel, W. Todd Lowther, Herman W. Odens, Jeffrey D. Schmitt.
Application Number | 20130345270 13/988707 |
Document ID | / |
Family ID | 46051251 |
Filed Date | 2013-12-26 |
United States Patent
Application |
20130345270 |
Kind Code |
A1 |
Kridel; Steven J. ; et
al. |
December 26, 2013 |
METHODS OF TREATING CANCER AND OTHER DISORDERS
Abstract
Active compounds useful for inhibiting fatty acid synthase in a
subject in need thereof are described. The active compounds are, in
general, a 5-mercapto-1H-Indazole-4,7-dione or an analog thereof.
The compounds are useful for treating subjects afflicted with,
cancer, obesity, diabetes, a viral infection, a bacterial
infection, a fungal infection, or a protozoal infection.
Inventors: |
Kridel; Steven J.;
(Clemmons, NC) ; Lowther; W. Todd; (Pfafftown,
NC) ; Odens; Herman W.; (Winston-Salem, NC) ;
Schmitt; Jeffrey D.; (Asheville, NC) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Kridel; Steven J.
Lowther; W. Todd
Odens; Herman W.
Schmitt; Jeffrey D. |
Clemmons
Pfafftown
Winston-Salem
Asheville |
NC
NC
NC
NC |
US
US
US
US |
|
|
Assignee: |
Waake Forest University Health
Sciences
Winston-Salem
NC
|
Family ID: |
46051251 |
Appl. No.: |
13/988707 |
Filed: |
November 7, 2011 |
PCT Filed: |
November 7, 2011 |
PCT NO: |
PCT/US11/59527 |
371 Date: |
September 9, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61412915 |
Nov 12, 2010 |
|
|
|
Current U.S.
Class: |
514/379 ;
514/405 |
Current CPC
Class: |
A61K 31/42 20130101;
C07D 261/20 20130101; C07D 231/56 20130101; A61K 31/416
20130101 |
Class at
Publication: |
514/379 ;
514/405 |
International
Class: |
C07D 231/56 20060101
C07D231/56; C07D 261/20 20060101 C07D261/20 |
Claims
1. A method of reversibly or irreversibly inhibiting fatty acid
synthase in a subject in need thereof, comprising administering to
said subject an active compound as described herein in a
treatment-effective amount; wherein said active compound is an
5-mercapto-1H-Indazole-4,7-dione or an analog thereof.
2. The method of claim 1, wherein said subject is afflicted with
cancer.
3. The method of claim 1, wherein said subject is afflicted with
obesity.
4. The method of claim 1, wherein said subject is afflicted with
diabetes
5. The method of claim 1, wherein said subject is afflicted with a
viral infection.
6. The method of claim 1, wherein said subject is afflicted with a
bacterial infection.
7. The method of claim 1, wherein said subject is afflicted with a
fungal infection.
8. The method of claim 1, wherein said subject is afflicted with a
protozoal infection.
9. The method of claim 1, wherein said active compound is selected
from the group consisting of compounds of Formula Ia, Ib and Ic, or
pharmaceutically acceptable salts or prodrugs thereof: ##STR00680##
where: each Z is independently selected from the group consisting
of N, O, S and CH.sub.2; A is O (then R.sub.3 is null) or N n=1-5;
m=1-5; j=1-4 E is N, CH or CH.sub.2; R.sub.1, R.sub.2, R.sub.3,
R.sub.4, and R.sub.5 individually can represent H, lower
straight-chain alkyl (e.g. alkyl groups containing one to ten
carbon atoms, such as methyl, ethyl or hexyl) or branched chain
lower alkyl, aromatic (e.g. phenyl, naphthyl) substituted aromatic,
heteroaromatic (e.g., pyridyl, pyrimidinyl, pyrazole, imidizole,
triazole, oxazole, isoxazole, thiazole, isothiazole, oxadiazole,
thiadiazole, pyridazine, triazine, indole, indazole,
benzoisoxazole, benzoxazole, benzoisothiazole, benzothiazole),
substituted heteroaromatic, benzyl, substituted benzyl. R.sub.1,
R.sub.2, R.sub.3, R.sub.4, and R.sub.5 individually can also
represent --(CH.sub.2).sub.n--Ar, where n is preferably 1 to 3 and
Ar is aromatic, substituted aromatic, heteroaromatic or substituted
heteroaromatic. A and B individually can also represent --OR, --SR,
--NR.sub.2 (where R individually represents H, straight chain or
branched chain lower alkoxy, aromatic, substituted aromatic,
heteroaromatic, substituted heteroaromatic), halogen, aldehyde,
carboxylic acid, --COOR, --CONHR, --CONR.sub.2, --OCONHR,
--OCONR.sub.2, --NCONHR, --NCONR.sub.2, --SCONR.sub.2. R.sub.1,
R.sub.2, R.sub.3, R.sub.4, and R.sub.5 can also be individually
selected from --(CH.sub.2).sub.n--R', (where R' individually
represents H, straight chain or branched chain lower alkoxy,
aromatic, substituted aromatic, heteroaromatic, substituted
heteroaromatic, halogen, aldehyde, carboxylic acid, --COOR,
--CONHR, --CONR.sub.2, --OCONHR, --OCONR.sub.2, --NCONHR,
--NCONR.sub.2, --SCONR.sub.2 (where R individually represents H,
straight chain or branched chain lower alkoxy, aromatic,
substituted aromatic, heteroaromatic, substituted heteroaromatic).
In some embodiments, R.sub.1, R.sub.2, R.sub.3, R.sub.4, and
R.sub.5 are each independently selected from the group consisting
of: H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl,
cycloalkylalkenyl, cycloalkylalkynyl, heterocyclo,
heterocycloalkyl, heterocycloalkenyl, heterocycloalkynyl, aryl,
arylalkyl, arylalkenyl, arylalkynyl, heteroaryl, heteroarylalkyl,
heteroarylalkenyl, heteroarylalkynyl, alkoxy, halo, mercapto,
azido, cyano, formyl, carboxylic acid, hydroxyl, nitro, acyl,
aryloxy, alkylthio, amino, alkylamino, arylalkylamino,
disubstituted amino, acylamino, acyloxy, ester, amide, sulfoxyl,
sulfonyl, sulfonate, sulfonic acid, sulfonamide, urea,
alkoxylacylamino, and aminoacyloxy.
10. The method of claim 1, wherein said active compound is selected
from the group consisting of the compounds of Table 1 below
TABLE-US-00003 TABLE 1 ##STR00681## ##STR00682## ##STR00683##
##STR00684## ##STR00685## ##STR00686## ##STR00687## ##STR00688##
##STR00689## ##STR00690## ##STR00691## ##STR00692## ##STR00693##
##STR00694## ##STR00695## ##STR00696## ##STR00697## ##STR00698##
##STR00699## ##STR00700## ##STR00701## ##STR00702## ##STR00703##
##STR00704## ##STR00705## ##STR00706## ##STR00707## ##STR00708##
##STR00709## ##STR00710## ##STR00711## ##STR00712## ##STR00713##
##STR00714## ##STR00715## ##STR00716## ##STR00717## ##STR00718##
##STR00719## ##STR00720## ##STR00721## ##STR00722## ##STR00723##
##STR00724## ##STR00725## ##STR00726## ##STR00727## ##STR00728##
##STR00729## ##STR00730## ##STR00731## ##STR00732## ##STR00733##
##STR00734## ##STR00735## ##STR00736## ##STR00737## ##STR00738##
##STR00739## ##STR00740## ##STR00741## ##STR00742## ##STR00743##
##STR00744## ##STR00745## ##STR00746## ##STR00747## ##STR00748##
##STR00749## ##STR00750## ##STR00751## ##STR00752## ##STR00753##
##STR00754## ##STR00755## ##STR00756## ##STR00757## ##STR00758##
##STR00759## ##STR00760## ##STR00761## ##STR00762## ##STR00763##
##STR00764## ##STR00765## ##STR00766## ##STR00767## ##STR00768##
##STR00769## ##STR00770## ##STR00771## ##STR00772## ##STR00773##
##STR00774## ##STR00775## ##STR00776## ##STR00777## ##STR00778##
##STR00779## ##STR00780## ##STR00781## ##STR00782## ##STR00783##
##STR00784## ##STR00785## ##STR00786## ##STR00787## ##STR00788##
##STR00789## ##STR00790## ##STR00791## ##STR00792## ##STR00793##
##STR00794## ##STR00795## ##STR00796## ##STR00797## ##STR00798##
##STR00799## ##STR00800## ##STR00801## ##STR00802##
##STR00803##
##STR00804## ##STR00805## ##STR00806## ##STR00807## ##STR00808##
##STR00809## ##STR00810## ##STR00811## ##STR00812## ##STR00813##
##STR00814## ##STR00815## ##STR00816## ##STR00817## ##STR00818##
##STR00819## ##STR00820## ##STR00821## ##STR00822## ##STR00823##
##STR00824## ##STR00825## ##STR00826## ##STR00827## ##STR00828##
##STR00829## ##STR00830## ##STR00831## ##STR00832## ##STR00833##
##STR00834## ##STR00835## ##STR00836## ##STR00837## ##STR00838##
##STR00839## ##STR00840## ##STR00841## ##STR00842## ##STR00843##
##STR00844## ##STR00845## ##STR00846## ##STR00847## ##STR00848##
##STR00849## ##STR00850## ##STR00851## ##STR00852## ##STR00853##
##STR00854## ##STR00855## ##STR00856## ##STR00857## ##STR00858##
##STR00859## ##STR00860## ##STR00861## ##STR00862## ##STR00863##
##STR00864## ##STR00865## ##STR00866## ##STR00867## ##STR00868##
##STR00869## ##STR00870## ##STR00871## ##STR00872## ##STR00873##
##STR00874## ##STR00875## ##STR00876## ##STR00877## ##STR00878##
##STR00879## ##STR00880## ##STR00881## ##STR00882## ##STR00883##
##STR00884## ##STR00885## ##STR00886## ##STR00887## ##STR00888##
##STR00889## ##STR00890## ##STR00891## ##STR00892## ##STR00893##
##STR00894## ##STR00895## ##STR00896## ##STR00897## ##STR00898##
##STR00899## ##STR00900## ##STR00901## ##STR00902## ##STR00903##
##STR00904## ##STR00905## ##STR00906## ##STR00907## ##STR00908##
##STR00909## ##STR00910## ##STR00911## ##STR00912## ##STR00913##
##STR00914## ##STR00915## ##STR00916## ##STR00917## ##STR00918##
##STR00919## ##STR00920## ##STR00921## ##STR00922## ##STR00923##
##STR00924## ##STR00925## ##STR00926## ##STR00927##
##STR00928##
##STR00929## ##STR00930## ##STR00931## ##STR00932## ##STR00933##
##STR00934## ##STR00935## ##STR00936## ##STR00937## ##STR00938##
##STR00939## ##STR00940## ##STR00941## ##STR00942## ##STR00943##
##STR00944## ##STR00945## ##STR00946## ##STR00947## ##STR00948##
##STR00949## ##STR00950## ##STR00951## ##STR00952## ##STR00953##
##STR00954## ##STR00955## ##STR00956## ##STR00957## ##STR00958##
##STR00959## ##STR00960## ##STR00961## ##STR00962## ##STR00963##
##STR00964## ##STR00965## ##STR00966## ##STR00967## ##STR00968##
##STR00969## ##STR00970## ##STR00971## ##STR00972## ##STR00973##
##STR00974## ##STR00975## ##STR00976## ##STR00977## ##STR00978##
##STR00979## ##STR00980## ##STR00981## ##STR00982## ##STR00983##
##STR00984## ##STR00985## ##STR00986## ##STR00987## ##STR00988##
##STR00989## ##STR00990## ##STR00991## ##STR00992## ##STR00993##
##STR00994## ##STR00995## ##STR00996## ##STR00997## ##STR00998##
##STR00999## ##STR01000## ##STR01001## ##STR01002## ##STR01003##
##STR01004## ##STR01005## ##STR01006## ##STR01007## ##STR01008##
##STR01009## ##STR01010## ##STR01011## ##STR01012## ##STR01013##
##STR01014## ##STR01015## ##STR01016## ##STR01017## ##STR01018##
##STR01019## ##STR01020## ##STR01021## ##STR01022## ##STR01023##
##STR01024## ##STR01025## ##STR01026## ##STR01027## ##STR01028##
##STR01029## ##STR01030## ##STR01031## ##STR01032## ##STR01033##
##STR01034## ##STR01035## ##STR01036## ##STR01037## ##STR01038##
##STR01039## ##STR01040## ##STR01041## ##STR01042## ##STR01043##
##STR01044## ##STR01045## ##STR01046## ##STR01047## ##STR01048##
##STR01049## ##STR01050## ##STR01051## ##STR01052## ##STR01053##
##STR01054##
##STR01055## ##STR01056## ##STR01057## ##STR01058## ##STR01059##
##STR01060## ##STR01061## ##STR01062## ##STR01063## ##STR01064##
##STR01065## ##STR01066## ##STR01067## ##STR01068## ##STR01069##
##STR01070## ##STR01071## ##STR01072## ##STR01073## ##STR01074##
##STR01075## ##STR01076## ##STR01077## ##STR01078## ##STR01079##
##STR01080## ##STR01081## ##STR01082## ##STR01083## ##STR01084##
##STR01085## ##STR01086## ##STR01087## ##STR01088## ##STR01089##
##STR01090## ##STR01091## ##STR01092## ##STR01093## ##STR01094##
##STR01095## ##STR01096## ##STR01097## ##STR01098## ##STR01099##
##STR01100## ##STR01101## ##STR01102## ##STR01103## ##STR01104##
##STR01105## ##STR01106## ##STR01107## ##STR01108## ##STR01109##
##STR01110## ##STR01111## ##STR01112## ##STR01113## ##STR01114##
##STR01115## ##STR01116## ##STR01117## ##STR01118## ##STR01119##
##STR01120## ##STR01121## ##STR01122## ##STR01123## ##STR01124##
##STR01125## ##STR01126## ##STR01127## ##STR01128## ##STR01129##
##STR01130## ##STR01131## ##STR01132## ##STR01133## ##STR01134##
##STR01135## ##STR01136## ##STR01137## ##STR01138## ##STR01139##
##STR01140## ##STR01141## ##STR01142## ##STR01143## ##STR01144##
##STR01145## ##STR01146## ##STR01147## ##STR01148## ##STR01149##
##STR01150## ##STR01151## ##STR01152## ##STR01153## ##STR01154##
##STR01155## ##STR01156## ##STR01157## ##STR01158## ##STR01159##
##STR01160## ##STR01161## ##STR01162## ##STR01163## ##STR01164##
##STR01165## ##STR01166## ##STR01167## ##STR01168## ##STR01169##
##STR01170## ##STR01171## ##STR01172## ##STR01173## ##STR01174##
##STR01175## ##STR01176## ##STR01177## ##STR01178##
##STR01179##
##STR01180## ##STR01181## ##STR01182## ##STR01183## ##STR01184##
##STR01185## ##STR01186## ##STR01187## ##STR01188## ##STR01189##
##STR01190## ##STR01191## ##STR01192## ##STR01193## ##STR01194##
##STR01195## ##STR01196## ##STR01197## ##STR01198## ##STR01199##
##STR01200## ##STR01201## ##STR01202## ##STR01203## ##STR01204##
##STR01205## ##STR01206## ##STR01207## ##STR01208## ##STR01209##
##STR01210## ##STR01211## ##STR01212## ##STR01213## ##STR01214##
##STR01215## ##STR01216## ##STR01217## ##STR01218## ##STR01219##
##STR01220## ##STR01221## ##STR01222## ##STR01223## ##STR01224##
##STR01225## ##STR01226## ##STR01227## ##STR01228## ##STR01229##
##STR01230## ##STR01231## ##STR01232## ##STR01233## ##STR01234##
##STR01235## ##STR01236## ##STR01237## ##STR01238## ##STR01239##
##STR01240## ##STR01241## ##STR01242## ##STR01243## ##STR01244##
##STR01245## ##STR01246## ##STR01247## ##STR01248## ##STR01249##
##STR01250## ##STR01251## ##STR01252## ##STR01253## ##STR01254##
##STR01255## ##STR01256## ##STR01257## ##STR01258## ##STR01259##
##STR01260## ##STR01261## ##STR01262## ##STR01263## ##STR01264##
##STR01265## ##STR01266## ##STR01267## ##STR01268## ##STR01269##
##STR01270## ##STR01271## ##STR01272## ##STR01273## ##STR01274##
##STR01275## ##STR01276## ##STR01277## ##STR01278## ##STR01279##
##STR01280## ##STR01281## ##STR01282## ##STR01283## ##STR01284##
##STR01285## ##STR01286## ##STR01287## ##STR01288## ##STR01289##
##STR01290## ##STR01291## ##STR01292## ##STR01293## ##STR01294##
##STR01295## ##STR01296## ##STR01297## ##STR01298## ##STR01299##
##STR01300## ##STR01301## ##STR01302## ##STR01303## ##STR01304##
##STR01305##
##STR01306## ##STR01307## ##STR01308## ##STR01309## ##STR01310##
##STR01311## ##STR01312## ##STR01313## ##STR01314## ##STR01315##
##STR01316## ##STR01317## ##STR01318## ##STR01319## ##STR01320##
##STR01321## ##STR01322## ##STR01323## ##STR01324## ##STR01325##
##STR01326## ##STR01327## ##STR01328## ##STR01329## ##STR01330##
##STR01331## ##STR01332## ##STR01333##
and pharmaceutically acceptable salts or prodrugs thereof.
11. (canceled)
Description
FIELD OF THE INVENTION
[0001] The present invention concerns methods of treatment, and
compounds and compositions useful for carrying out such
methods.
SUMMARY OF THE INVENTION
[0002] A first aspect of the present invention is a method of
inhibiting fatty acid synthase in a subject in need thereof,
comprising administering to said subject an active compound as
described herein in a treatment-effective amount. Examples of
subjects in need thereof include, but are not limited to, subjects
afflicted with cancer, obesity, diabetes, viral infection,
bacterial infection, fungal infection or protozoal infection.
[0003] A second aspect of the present invention is the use of an
active compound as described herein for the preparation of a
medicament for carrying out a method as described herein, such as
for treating cancer, obesity, diabetes, viral infection, bacterial
infection, fungal infection or protozoal infection.
[0004] A third aspect of the present invention is the use of an
active compound as described herein for carrying out a method as
described herein, such as for treating cancer, obesity, diabetes,
viral infection, bacterial infection, fungal infection or protozoal
infection.
[0005] A further aspect of the present invention is the use of an
active compound, as described herein, in combination with one or
more known medicaments to achieve additive or synergistic activity
for treating cancer, obesity, diabetes, viral infection, bacterial
infection, fungal infection or protozoal infection.
[0006] The foregoing and other objects and aspects of the present
invention are explained in greater detail in the specification set
forth below.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
[0007] The present invention is primarily concerned with the
treatment of human subjects, but the invention may also be carried
out on animal subjects, particularly mammalian subjects such as
dogs, cats, livestock and horses for veterinary purposes. Human
subjects may be male or female subjects and may be any age,
including neonate, infant, juvenile, adolescent, adult, and
geriatric subjects.
[0008] "Treat" as used herein refers to any type of treatment that
imparts a benefit to a patient, particularly delaying or retarding
the progression of the disease.
[0009] "Pharmaceutically acceptable" as used herein means that the
compound or composition is suitable for administration to a subject
to achieve the treatments described herein, without unduly
deleterious side effects in light of the severity of the disease
and necessity of the treatment.
[0010] "Concurrently" as used herein means sufficiently close in
time to produce a combined effect (that is, concurrently may be
simultaneously, or it may be two or more events occurring within a
short time period before or after each other).
[0011] "Cancer" as used herein includes, but is not limited to, any
cancers exhibiting elevated levels of fatty acid synthase as in
epithelial cancers; the term "cancer" includes also breast cancer,
lung cancer, prostate cancer, ovarian cancer, colon or colorectal
cancer, liver cancer, skin cancer such as melanoma, brain cancer
such as astrocytoma, pancreatic cancer, leukemia, lymphoma,
etc.
[0012] "Diabetes" as used herein includes type I and type II
diabetes.
[0013] "Bacterial infection" as used herein refers to any undesired
infection of a subject with a gram negative or gram positive
bacteria. Particular examples are infections by those bacteria set
forth below.
[0014] "Gram-negative" bacteria are those that do not retain
crystal violet dye after an alcohol wash in the Gram staining
protocol. This is due to structural properties in the cell walls of
the bacteria. Many genera and species of Gram-negative bacteria are
pathogenic. Gram-negative bacteria include members of the phylum
proteobacteria, which include genus members Escherichia,
Salmonella, Vibrio, and Helicobacter. Examples of genera of
biofilm-forming bacteria include, but are not limited to,
Pseudomonas, Bordetella, Vibrio, Haemophilus, Halomonas, and
Acinetobacter. Other genera include Klebsiella, Proteus, Neisseria,
Helicobacter, Brucella, Legionella, Campylobacter, Francisella,
Pasteurella, Yersinia, Bartonella, Bacteroides, Streptobacillus,
Spirillum, Moraxella and Shigella. Examples of species of bacteria
include Pseudomonas aeuroginosa, Bordetella pertussis, Vibrio
vulnificus, Haemophilus influenzae, and Halomonas pacifica.
[0015] Gram-negative bacteria of the Acinetobacter genus belong to
the phylum Gammaproteobacteria, order Pseudomonadalas, and family
Moraxellaceae. Genus members include, but are not limited to,
Acinetobacter bumannii, Acinetobacter haemolyticus, and
Acinetobacter lwoffi. Various nosocomial infections that are
especially prevalent in intensive care units implicate
Acinetobacter species such as Acinetobacter baumannii and
Acinetobacter lwoffi. Acinetobacter baumanni is a frequent cause of
nosocomial pneumonia, and can also cause skin and wound infections
and bacteremia. Acinetobacter lwoffi causes meningitis. The
Acinetobacter species are resistant to many classes of
antibiotics.
[0016] Examples of Gram-positive bacteria include, but are not
limited to, bacteria of the genera Listeria, Staphylococcus,
Streptococcus, Bacillus, Corynebacterium, Peptostreptococcus, and
Clostridium. Species include, but not limited to, Listeria
monocytogenes, Staphylococcus aureus, Streptococcus pyogenes,
Streptococcus pneumoniae, Bacillus cereus, Bacillus anthracis,
Clostridium botulinum, Clostridium perfringens, Clostridium
difficile, Clostridium tetani, Corynebacterium diphtheriae,
Corynebacterium ulcerans, and Peptostreptococcus anaerobius. Other
bacterial genera include, but are not limited to, Actinomyces,
Propionibacterium, Nocardia and Streptomyces. Staphylococcus aureus
is a common cause of nosocomial infections, often found in
post-surgical wound infections. Staphylococcus aureus can also
cause a variety of other infections in humans (e.g., skin
infections), as well as contribute to mastitis in dairy cows.
Methicillin-resistant Staphylococcus aureaus (MRSA), in particular,
is especially difficult to treat due to multiple drug resistances,
including penicillins and cephalosporins. MRSA has become
problematic in hospital settings, particularly among the more
susceptible patients with open wounds, invasive devices, weakened
immune systems, etc.
[0017] "Fungal infection" as used herein refers to any undesired
infection of a subject with fungal cells, including but not limited
to infections with Aspergillus, Candida, Cryptococcus,
Coccidioides, Tinea, Sporothrix, Blastomyces, Histoplasma,
Pneumocystis and Saccharomyces. Additionally, fungal cells include,
but is not limited to, Aspergillus fumigatus, Aspergillus flavus,
Aspergillus niger, Aspergillus terreus, Aspergillus nidulans,
Candida albicans, Coccidioides immitis, Cryptococcus neoformans,
Tinea unguium, Tinea corporis, Tinea cruris, Sporothrix schenckii,
Blastomyces dermatitidis, Histoplasma capsulatum, Histoplasma
duboisii, and Saccharomyces cerevisiae.
[0018] "Protozoal infection" as used herein refers to any undesired
infection of a subject with protozoal cells, including but not
limited to infection with Leishmania, Kokzidioa, Trypanosoma,
Chlamydia, and Rickettsia, and more particularly infection with
Histoplasma capsulatum, Entamoeba histolytica, Trichomonas tenas,
Trichomonas hominis, Trichomonas vaginalis, Trypanosoma gambiense,
Trypanosoma rhodesiense, Trypanosoma cruzi, Leishmania donovani,
Leishmania tropica, Leishmania braziliensis, Pneumocystis
pneumonia, Enterobius vermicularis, Trichuris trichiura, Ascaris
lumbricoides, Trichinella spiralis, Strongyloides stercoralis,
Schistosoma japonicum, Schistosoma mansoni, Schistosoma
haematobium, etc.
[0019] "Viral infection" as used herein includes but is not limited
to such as severe acute respiratory syndrome; coronavirus
infection; influenza infection; SARS virus infection; an
orthomyxovirus-coronavirus hybrid infection; Hepatitis C infection,
RSV infection, etc.
[0020] "Obesity" as used herein refers not only to morbid obesity
but any undesired elevation in weight or associated condition which
may be the subject of medical treatment. (see, e.g., Sylvia
Lee-Huang et al., US Patent Application No. 20090061031 (Mar. 5,
2009). Thus the treatment of obesity as described herein includes
methods of treating, controlling or preventing obesity, or of
reducing body weight, or of inhibiting fat accumulation, or of
promoting fat burning and energy uncoupling in vivo, or of
treating, controlling or preventing the onset of one or more
obesity-related disorders or conditions, comprising administering
an active agent as described herein. Obesity-related disorders or
conditions that may be treated by the methods of the present
invention include, but are not limited to, coronary artery disease,
hypertension, stroke, peripheral vascular disease, insulin
resistance, glucose intolerance, diabetes mellitus, hyperglycemia,
hyperlipidemia, hypercholesteremia, hypertriglyceridemia,
hyperinsulinemia, atherosclerosis, cellular proliferation and
endothelial dysfunction, diabetic dyslipidemia, HIV-related
lipodystrophy and metabolic syndrome, type II diabetes, diabetic
complications including diabetic neuropathy, nephropathy,
retinopathy or cataracts, heart failure, inflammation, thrombosis,
congestive heart failure, any other asthmatic or pulmonary disease
related to obesity and any other viral infection, infection related
diseases and any other cardiovascular disease related to obesity or
an overweight condition.
[0021] "Alkyl" as used herein alone or as part of another group,
refers to a straight or branched chain hydrocarbon containing from
1 to 10 carbon atoms. Representative examples of alkyl include, but
are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl,
sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl,
n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl,
n-heptyl, n-octyl, n-nonyl, n-decyl, and the like. Alkyl groups as
used herein may be lower alkyl. "Lower alkyl" as used herein, is a
subset of alkyl, in some embodiments preferred, and refers to a
straight or branched chain hydrocarbon group containing from 1 to 4
carbon atoms. Representative examples of lower alkyl include, but
are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl,
iso-butyl, tert-butyl, and the like.
[0022] The term "alkyl" or "loweralkyl" is intended to include both
substituted and unsubstituted alkyl or loweralkyl unless otherwise
indicated and these groups may be substituted with groups selected
from halo (e.g., haloalkyl), alkyl, haloalkyl, alkenyl, alkynyl,
cycloalkyl, cycloalkylallyl, aryl, arylalkyl, heterocyclo,
heterocycloalkyl, hydroxyl, alkoxy (thereby creating a polyalkoxy
such as polyethylene glycol), alkenyloxy, alkynyloxy, haloalkoxy,
cycloalkoxy, cycloalkylalkyloxy, aryloxy, arylalkyloxy,
heterocyclooxy, heterocyclolalkyloxy, mercapto, alkyl-S(O).sub.m,
haloalkyl-S(O).sub.m, alkenyl-S(O).sub.m, alkynyl-S(O).sub.m,
cycloalkyl-S(O).sub.m, cycloalkylalkyl-S(O).sub.m, aryl-S(O).sub.m,
arylalkyl-S(O).sub.m, heterocyclo-S(O).sub.m,
heterocycloalkyl-S(O).sub.m, amino, carboxy, alkylamino,
alkenylamino, alkynylamino, haloalkylamino, cycloalkylamino,
cycloalkylalkylamino, arylamino, arylalkylamino, heterocycloamino,
heterocycloalkylamino, disubstituted-amino, acylamino, acyloxy,
ester, amide, sulfonamide, urea, alkoxyacylamino, aminoacyloxy,
nitro or cyano where m=0, 1, 2 or 3.
[0023] "Alkenyl" as used herein alone or as part of another group,
refers to a straight or branched chain hydrocarbon containing from
1 to 10 carbon atoms (or in loweralkenyl 1 to 4 carbon atoms) which
include 1 to 4 double bonds in the normal chain. Representative
examples of alkenyl include, but are not limited to, vinyl,
2-propenyl, 3-butenyl, 2-butenyl, 4-pentenyl, 3-pentenyl,
2-hexenyl, 3-hexenyl, 2,4-heptadiene, and the like. The term
"alkenyl" or "loweralkenyl" is intended to include both substituted
and unsubstituted alkenyl or loweralkenyl unless otherwise
indicated and these groups may be substituted with groups as
described in connection with alkyl and loweralkyl above.
[0024] "Alkynyl" as used herein alone or as part of another group,
refers to a straight or branched chain hydrocarbon containing from
1 to 10 carbon atoms (or in loweralkynyl 1 to 4 carbon atoms) which
include 1 triple bond in the normal chain. Representative examples
of alkynyl include, but are not limited to, 2-propynyl, 3-butynyl,
2-butynyl, 4-pentynyl, 3-pentynyl, and the like. The term "alkynyl"
or "loweralkynyl" is intended to include both substituted and
unsubstituted alkynyl or loweralknynyl unless otherwise indicated
and these groups may be substituted with the same groups as set
forth in connection with alkyl and loweralkyl above.
[0025] "Cycloalkyl" as used herein alone or as part of another
group, refers to a saturated or partially unsaturated cyclic
hydrocarbon group containing from 3, 4 or 5 to 6, 7 or 8 carbons
(which carbons may be replaced in a heterocyclic group as discussed
below). Representative examples of cycloalkyl include, cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
These rings may be optionally substituted with additional
substituents as described herein such as halo or loweralkyl. The
term "cycloalkyl" is generic and intended to include heterocyclic
groups as discussed below unless specified otherwise.
[0026] "Heterocyclic group" or "heterocyclo" as used herein alone
or as part of another group, refers to an aliphatic (e.g., fully or
partially saturated heterocyclo) or aromatic (e.g., heteroaryl)
monocyclic- or a bicyclic-ring system. Monocyclic ring systems are
exemplified by any 5 or 6 membered ring containing 1, 2, 3, or 4
heteroatoms independently selected from oxygen, nitrogen and
sulfur. The 5 membered ring has from 0-2 double bonds and the 6
membered ring has from 0-3 double bonds. Representative examples of
monocyclic ring systems include, but are not limited to, azetidine,
azepine, aziridine, diazepine, 1,3-dioxolane, dioxane, dithiane,
furan, imidazole, imidazoline, imidazolidine, isothiazole,
isothiazoline, isothiazolidine, isoxazole, isoxazoline,
isoxazolidine, morpholine, oxadiazole, oxadiazoline,
oxadiazolidine, oxazole, oxazoline, oxazolidine, piperazine,
piperidine, pyran, pyrazine, pyrazole, pyrazoline, pyrazolidine,
pyridine, pyrimidine, pyridazine, pyrrole, pyrroline, pyrrolidine,
tetrahydrofuran, tetrahydrothiophene, tetrazine, tetrazole,
thiadiazole, thiadiazoline, thiadiazolidine, thiazole, thiazoline,
thiazolidine, thiophene, thiomorpholine, thiomorpholine sulfone,
thiopyran, triazine, triazole, trithiane, and the like. Bicyclic
ring systems are exemplified by any of the above monocyclic ring
systems fused to an aryl group as defined herein, a cycloalkyl
group as defined herein, or another monocyclic ring system as
defined herein. Representative examples of bicyclic ring systems
include but are not limited to, for example, benzimidazole,
benzothiazole, benzothiadiazole, benzothiophene, benzoxadiazole,
benzoxazole, benzofuran, benzopyran, benzothiopyran, benzodioxine,
1,3-benzodioxole, cinnoline, indazole, indole, indoline,
indolizine, naphthyridine, isobenzofuran, isobenzothiophene,
isoindole, isoindoline, isoquinoline, phthalazine, purine,
pyranopyridine, quinoline, quinolizine, quinoxaline, quinazoline,
tetrahydroisoquinoline, tetrahydroquinoline, thiopyranopyridine,
and the like. These rings include quaternized derivatives thereof
and may be optionally substituted with groups selected from halo,
alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl,
aryl, arylalkyl, heterocyclo, heterocycloalkyl, hydroxyl, alkoxy,
alkenyloxy, alkynyloxy, haloalkoxy, cycloalkoxy,
cycloalkylalkyloxy, aryloxy, arylalkyloxy, heterocyclooxy,
heterocyclolalkyloxy, mercapto, alkyl-S(O).sub.m, haloalkyl-S(O),
alkenyl-S(O).sub.m, heterocyclo-S(O).sub.m,
heterocycloalkyl-S(O).sub.m, amino, alkylamino, alkenylamino,
alkynylamino, haloalkylamino, cycloalkylamino,
cycloalkylalkylamino, arylamino, arylalkylamino, heterocycloamino,
heterocycloalkylamino, disubstituted-amino, acylamino, acyloxy,
ester, amide, sulfonamide, urea, alkoxyacylamino, aminoacyloxy,
nitro or cyano where m=0, 1, 2 or 3.
[0027] "Aryl" as used herein alone or as part of another group,
refers to a monocyclic carbocyclic ring system or a bicyclic
carbocyclic fused ring system having one or more aromatic rings.
Representative examples of aryl include, azulenyl, indanyl,
indenyl, naphthyl, phenyl, tetrahydronaphthyl, and the like. The
term "aryl" is intended to include both substituted and
unsubstituted aryl unless otherwise indicated and these groups may
be substituted with the same groups as set forth in connection with
alkyl and loweralkyl above.
[0028] "Arylalkyl" as used herein alone or as part of another
group, refers to an aryl group, as defined herein, appended to the
parent molecular moiety through an alkyl group, as defined herein.
Representative examples of arylalkyl include, but are not limited
to, benzyl, 2-phenylethyl, 3-phenylpropyl, 2-naphth-2-ylethyl, and
the like.
[0029] "Heteroaryl" as used herein is as described in connection
with heterocyclo above.
[0030] "Alkoxy" as used herein alone or as part of another group,
refers to an alkyl or loweralkyl group, as defined herein (and thus
including substituted versions such as polyalkoxy), appended to the
parent molecular moiety through an oxy group, --O--. Representative
examples of alkoxy include, but are not limited to, methoxy,
ethoxy, propoxy, 2-propoxy, butoxy, tert-butoxy, pentyloxy,
hexyloxy and the like.
[0031] "Halo" as used herein refers to any suitable halogen,
including --F, --Cl, --Br, and --I.
[0032] "Mercapto" as used herein refers to an --SH group.
[0033] "Azido" as used herein refers to an --N.sub.3 group.
[0034] "Cyano" as used herein refers to a --CN group.
[0035] "Formyl" as used herein refers to a --C(O)H group.
[0036] "Carboxylic acid" as used herein refers to a --C(O)OH
group.
[0037] "Hydroxyl" as used herein refers to an --OH group.
[0038] "Nitro" as used herein refers to an --NO.sub.2 group.
[0039] "Acyl" as used herein alone or as part of another group
refers to a --C(O)R radical,
where R is any suitable substituent such as aryl, alkyl, alkenyl,
alkynyl, cycloalkyl or other suitable substituent as described
herein.
[0040] "Alkylthio" as used herein alone or as part of another
group, refers to an alkyl group, as defined herein, appended to the
parent molecular moiety through a thio or mercapto moiety, as
defined herein. Representative examples of alkylthio include, but
are not limited, methylthio, ethylthio, tert-butylthio, hexylthio,
and the like.
[0041] "Amino" as used herein means the radical --NH.sub.2.
[0042] "Alkylamino" as used herein alone or as part of another
group means the radical --NHR, where R is an alkyl group.
[0043] "Arylalkylamino" as used herein alone or as part of another
group means the radical --NHR, where R is an arylalkyl group.
[0044] "Disubstituted-amino" as used herein alone or as part of
another group means the radical --NR.sub.aR.sub.b, where R.sub.a
and R.sub.b are independently selected from the groups alkyl,
haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl,
arylalkyl, heterocyclo, heterocycloalkyl.
[0045] "Acylamino" as used herein alone or as part of another group
means the radical --NR.sub.aR.sub.b, where R.sub.a is an acyl group
as defined herein and R.sub.b is selected from the groups hydrogen,
alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl,
aryl, arylalkyl, heterocyclo, heterocycloalkyl.
[0046] "Acyloxy" as used herein alone or as part of another group
means the radical --OR, where R is an acyl group as defined
herein.
[0047] "Ester" as used herein alone or as part of another group
refers to a --C(O)OR radical, where R is any suitable substituent
such as alkyl, cycloalkyl, alkenyl, alkynyl or aryl.
[0048] "Amide" as used herein alone or as part of another group
refers to a --C(O)NR.sub.aR.sub.b radical, where R.sub.a and
R.sub.b are any suitable substituent such as alkyl, cycloalkyl,
alkenyl, alkynyl or aryl.
[0049] "Sulfoxyl" as used herein refers to a compound of the
formula --S(O)R, where R is any suitable substituent such as alkyl,
cycloalkyl, alkenyl, alkynyl or aryl.
[0050] "Sulfonyl" as used herein refers to a compound of the
formula --S(O)(O)R, where R is any suitable substituent such as
alkyl, cycloalkyl, alkenyl, alkynyl or aryl.
[0051] "Sulfonate" as used herein refers to a compound of the
formula --S(O)(O)OR, where R is any suitable substituent such as
alkyl, cycloalkyl, alkenyl, alkynyl or aryl.
[0052] "Sulfonic acid" as used herein refers to a compound of the
formula --S(O)(O)OH.
[0053] "Sulfonamide" as used herein alone or as part of another
group refers to a --S(O).sub.2NR.sub.aR.sub.b radical, where
R.sub.a and R.sub.b are any suitable substituent such as H, alkyl,
cycloalkyl, alkenyl, alkynyl or aryl.
[0054] "Thio" and "mercapto" as used herein refers to a compound of
the formula --SR, where R is any suitable substituent such as
alkyl, cycloalkyl, alkenyl, alkynyl or aryl.
[0055] "Urea" as used herein alone or as part of another group
refers to an --N(R.sub.e)C(O)NR.sub.aR.sub.b radical, where
R.sub.a, R.sub.b and R.sub.e are any suitable substituent such as
H, alkyl, cycloalkyl, alkenyl, alkynyl or aryl.
[0056] "Alkoxyacylamino" as used herein alone or as part of another
group refers to an --N(R.sub.a)C(O)OR.sub.b radical, where R.sub.a,
R.sub.b are any suitable substituent such as H, alkyl, cycloalkyl,
alkenyl, alkynyl or aryl.
[0057] "Aminoacyloxy" as used herein alone or as part of another
group refers to an --OC(O)NR.sub.aR.sub.b radical, where R.sub.a
and R.sub.b are any suitable substituent such as H, alkyl,
cycloalkyl, alkenyl, alkynyl or aryl.
1. Active Compounds.
[0058] Active compounds of the present invention are, in general,
fatty acid synthase (FASN) inhibitors and analogs thereof,
particularly compounds that bind specifically or stereospecifically
to the thioesterase domain of mammalian (e.g., human) FASN.
Examples of such compounds include compounds of Formula Ia, Ib and
Ic:
##STR00001##
where:
[0059] each Z is independently selected from the group consisting
of N, O, S and CH.sub.2;
[0060] A is O (then R.sub.3 is null) or N
[0061] n=1-5; m=1-5; j=1-4
[0062] E is N, CH or CH.sub.2;
[0063] R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5 individually
can represent H, lower straight-chain alkyl (e.g. alkyl groups
containing one to ten carbon atoms, such as methyl, ethyl or hexyl)
or branched chain lower alkyl, aromatic (e.g. phenyl, naphthyl)
substituted aromatic, heteroaromatic (e.g., pyridyl, pyrimidinyl,
pyrazole, imidizole, triazole, oxazole, isoxazole, thiazole,
isothiazole, oxadiazole, thiadiazole, pyridazine, triazine, indole,
indazole, benzoisoxazole, benzoxazole, benzoisothiazole,
benzothiazole), substituted heteroaromatic, benzyl, substituted
benzyl.
[0064] R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5 individually
can also represent --(CH.sub.2).sub.n--Ar, where n is preferably 1
to 3 and Ar is aromatic, substituted aromatic, heteroaromatic or
substituted heteroaromatic. A and B individually can also represent
--OR, --SR, --NR.sub.2 (where R individually represents H, straight
chain or branched chain lower alkoxy, aromatic, substituted
aromatic, heteroaromatic, substituted heteroaromatic), halogen,
aldehyde, carboxylic acid, --COOR, --CONHR, --CONR.sub.2, --OCONHR,
--OCONR.sub.2, --NCONHR, --NCONR.sub.2, --SCONR.sub.2.
[0065] R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5 can also be
individually selected from --(CH.sub.2).sub.nR', (where R'
individually represents H, straight chain or branched chain lower
alkoxy, aromatic, substituted aromatic, heteroaromatic, substituted
heteroaromatic, halogen, aldehyde, carboxylic acid, --COOR,
--CONHR, --CONR.sub.2, --OCONHR, --OCONR.sub.2, --NCONHR,
--NCONR.sub.2, --SCONR.sub.2 (where R individually represents H,
straight chain or branched chain lower alkoxy, aromatic,
substituted aromatic, heteroaromatic, substituted
heteroaromatic).
[0066] In some embodiments, R.sub.1, R.sub.2, R.sub.3, R.sub.4, and
R.sub.5 are each independently selected from the group consisting
of: H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl,
cycloalkylalkenyl, cycloalkylalkynyl, heterocyclo,
heterocycloalkyl, heterocycloalkenyl; heterocycloalkynyl, aryl,
arylalkyl, arylalkenyl, arylalkynyl, heteroaryl, heteroarylalkyl,
heteroarylalkenyl, heteroarylalkynyl, alkoxy, halo, mercapto,
azido, cyano, formyl, carboxylic acid, hydroxyl, nitro, acyl,
aryloxy, alkylthio, amino, alkylamino, arylalkylamino,
disubstituted amino, acylamino, acyloxy, ester, amide, sulfoxyl,
sulfonyl, sulfonate, sulfonic acid, sulfonamide, urea,
alkoxylacylamino, and aminoacyloxy.
[0067] In other embodiments R.sub.1, R.sub.2, R.sub.3, R.sub.4, and
R.sub.5 can also be pantetheine or a pantetheine derivative;
examples include, but are not limited to the foregoing examples in
formulas IIa, IIb and IIc:
##STR00002##
where:
[0068] p=1-5; m=0-5; r=1-8;
[0069] the asterisk to left of the structures above denotes the
attachment point.
Particular examples of active compounds of the present invention
include, but are not limited to, those set forth in the following
Table 1, and pharmaceutically acceptable salts or prodrugs
thereof.
TABLE-US-00001 TABLE 1 Example Active Compounds ##STR00003##
##STR00004## ##STR00005## ##STR00006## ##STR00007## ##STR00008##
##STR00009## ##STR00010## ##STR00011## ##STR00012## ##STR00013##
##STR00014## ##STR00015## ##STR00016## ##STR00017## ##STR00018##
##STR00019## ##STR00020## ##STR00021## ##STR00022## ##STR00023##
##STR00024## ##STR00025## ##STR00026## ##STR00027## ##STR00028##
##STR00029## ##STR00030## ##STR00031## ##STR00032## ##STR00033##
##STR00034## ##STR00035## ##STR00036## ##STR00037## ##STR00038##
##STR00039## ##STR00040## ##STR00041## ##STR00042## ##STR00043##
##STR00044## ##STR00045## ##STR00046## ##STR00047## ##STR00048##
##STR00049## ##STR00050## ##STR00051## ##STR00052## ##STR00053##
##STR00054## ##STR00055## ##STR00056## ##STR00057## ##STR00058##
##STR00059## ##STR00060## ##STR00061## ##STR00062## ##STR00063##
##STR00064## ##STR00065## ##STR00066## ##STR00067## ##STR00068##
##STR00069## ##STR00070## ##STR00071## ##STR00072## ##STR00073##
##STR00074## ##STR00075## ##STR00076## ##STR00077## ##STR00078##
##STR00079## ##STR00080## ##STR00081## ##STR00082## ##STR00083##
##STR00084## ##STR00085## ##STR00086## ##STR00087## ##STR00088##
##STR00089## ##STR00090## ##STR00091## ##STR00092## ##STR00093##
##STR00094## ##STR00095## ##STR00096## ##STR00097## ##STR00098##
##STR00099## ##STR00100## ##STR00101## ##STR00102## ##STR00103##
##STR00104## ##STR00105## ##STR00106## ##STR00107## ##STR00108##
##STR00109## ##STR00110## ##STR00111## ##STR00112## ##STR00113##
##STR00114## ##STR00115## ##STR00116## ##STR00117## ##STR00118##
##STR00119## ##STR00120## ##STR00121## ##STR00122## ##STR00123##
##STR00124## ##STR00125##
##STR00126## ##STR00127## ##STR00128## ##STR00129## ##STR00130##
##STR00131## ##STR00132## ##STR00133## ##STR00134## ##STR00135##
##STR00136## ##STR00137## ##STR00138## ##STR00139## ##STR00140##
##STR00141## ##STR00142## ##STR00143## ##STR00144## ##STR00145##
##STR00146## ##STR00147## ##STR00148## ##STR00149## ##STR00150##
##STR00151## ##STR00152## ##STR00153## ##STR00154## ##STR00155##
##STR00156## ##STR00157## ##STR00158## ##STR00159## ##STR00160##
##STR00161## ##STR00162## ##STR00163## ##STR00164## ##STR00165##
##STR00166## ##STR00167## ##STR00168## ##STR00169## ##STR00170##
##STR00171## ##STR00172## ##STR00173## ##STR00174## ##STR00175##
##STR00176## ##STR00177## ##STR00178## ##STR00179## ##STR00180##
##STR00181## ##STR00182## ##STR00183## ##STR00184## ##STR00185##
##STR00186## ##STR00187## ##STR00188## ##STR00189## ##STR00190##
##STR00191## ##STR00192## ##STR00193## ##STR00194## ##STR00195##
##STR00196## ##STR00197## ##STR00198## ##STR00199## ##STR00200##
##STR00201## ##STR00202## ##STR00203## ##STR00204## ##STR00205##
##STR00206## ##STR00207## ##STR00208## ##STR00209## ##STR00210##
##STR00211## ##STR00212## ##STR00213## ##STR00214## ##STR00215##
##STR00216## ##STR00217## ##STR00218## ##STR00219## ##STR00220##
##STR00221## ##STR00222## ##STR00223## ##STR00224## ##STR00225##
##STR00226## ##STR00227## ##STR00228## ##STR00229## ##STR00230##
##STR00231## ##STR00232## ##STR00233## ##STR00234## ##STR00235##
##STR00236## ##STR00237## ##STR00238## ##STR00239## ##STR00240##
##STR00241## ##STR00242## ##STR00243## ##STR00244## ##STR00245##
##STR00246## ##STR00247## ##STR00248## ##STR00249## ##STR00250##
##STR00251##
##STR00252## ##STR00253## ##STR00254## ##STR00255## ##STR00256##
##STR00257## ##STR00258## ##STR00259## ##STR00260## ##STR00261##
##STR00262## ##STR00263## ##STR00264## ##STR00265## ##STR00266##
##STR00267## ##STR00268## ##STR00269## ##STR00270## ##STR00271##
##STR00272## ##STR00273## ##STR00274## ##STR00275## ##STR00276##
##STR00277## ##STR00278## ##STR00279## ##STR00280## ##STR00281##
##STR00282## ##STR00283## ##STR00284## ##STR00285## ##STR00286##
##STR00287## ##STR00288## ##STR00289## ##STR00290## ##STR00291##
##STR00292## ##STR00293## ##STR00294## ##STR00295## ##STR00296##
##STR00297## ##STR00298## ##STR00299## ##STR00300## ##STR00301##
##STR00302## ##STR00303## ##STR00304## ##STR00305## ##STR00306##
##STR00307## ##STR00308## ##STR00309## ##STR00310## ##STR00311##
##STR00312## ##STR00313## ##STR00314## ##STR00315## ##STR00316##
##STR00317## ##STR00318## ##STR00319## ##STR00320## ##STR00321##
##STR00322## ##STR00323## ##STR00324## ##STR00325## ##STR00326##
##STR00327## ##STR00328## ##STR00329## ##STR00330## ##STR00331##
##STR00332## ##STR00333## ##STR00334## ##STR00335## ##STR00336##
##STR00337## ##STR00338## ##STR00339## ##STR00340## ##STR00341##
##STR00342## ##STR00343## ##STR00344## ##STR00345## ##STR00346##
##STR00347## ##STR00348## ##STR00349## ##STR00350## ##STR00351##
##STR00352## ##STR00353## ##STR00354## ##STR00355## ##STR00356##
##STR00357## ##STR00358## ##STR00359## ##STR00360## ##STR00361##
##STR00362## ##STR00363## ##STR00364## ##STR00365## ##STR00366##
##STR00367## ##STR00368## ##STR00369## ##STR00370## ##STR00371##
##STR00372## ##STR00373## ##STR00374## ##STR00375##
##STR00376##
##STR00377## ##STR00378## ##STR00379## ##STR00380## ##STR00381##
##STR00382## ##STR00383## ##STR00384## ##STR00385## ##STR00386##
##STR00387## ##STR00388## ##STR00389## ##STR00390## ##STR00391##
##STR00392## ##STR00393## ##STR00394## ##STR00395## ##STR00396##
##STR00397## ##STR00398## ##STR00399## ##STR00400## ##STR00401##
##STR00402## ##STR00403## ##STR00404## ##STR00405## ##STR00406##
##STR00407## ##STR00408## ##STR00409## ##STR00410## ##STR00411##
##STR00412## ##STR00413## ##STR00414## ##STR00415## ##STR00416##
##STR00417## ##STR00418## ##STR00419## ##STR00420## ##STR00421##
##STR00422## ##STR00423## ##STR00424## ##STR00425## ##STR00426##
##STR00427## ##STR00428## ##STR00429## ##STR00430## ##STR00431##
##STR00432## ##STR00433## ##STR00434## ##STR00435## ##STR00436##
##STR00437## ##STR00438## ##STR00439## ##STR00440## ##STR00441##
##STR00442## ##STR00443## ##STR00444## ##STR00445## ##STR00446##
##STR00447## ##STR00448## ##STR00449## ##STR00450## ##STR00451##
##STR00452## ##STR00453## ##STR00454## ##STR00455## ##STR00456##
##STR00457## ##STR00458## ##STR00459## ##STR00460## ##STR00461##
##STR00462## ##STR00463## ##STR00464## ##STR00465## ##STR00466##
##STR00467## ##STR00468## ##STR00469## ##STR00470## ##STR00471##
##STR00472## ##STR00473## ##STR00474## ##STR00475## ##STR00476##
##STR00477## ##STR00478## ##STR00479## ##STR00480## ##STR00481##
##STR00482## ##STR00483## ##STR00484## ##STR00485## ##STR00486##
##STR00487## ##STR00488## ##STR00489## ##STR00490## ##STR00491##
##STR00492## ##STR00493## ##STR00494## ##STR00495## ##STR00496##
##STR00497## ##STR00498## ##STR00499## ##STR00500## ##STR00501##
##STR00502##
##STR00503## ##STR00504## ##STR00505## ##STR00506## ##STR00507##
##STR00508## ##STR00509## ##STR00510## ##STR00511## ##STR00512##
##STR00513## ##STR00514## ##STR00515## ##STR00516## ##STR00517##
##STR00518## ##STR00519## ##STR00520## ##STR00521## ##STR00522##
##STR00523## ##STR00524## ##STR00525## ##STR00526## ##STR00527##
##STR00528## ##STR00529## ##STR00530## ##STR00531## ##STR00532##
##STR00533## ##STR00534## ##STR00535## ##STR00536## ##STR00537##
##STR00538## ##STR00539## ##STR00540## ##STR00541## ##STR00542##
##STR00543## ##STR00544## ##STR00545## ##STR00546## ##STR00547##
##STR00548## ##STR00549## ##STR00550## ##STR00551## ##STR00552##
##STR00553## ##STR00554## ##STR00555## ##STR00556## ##STR00557##
##STR00558## ##STR00559## ##STR00560## ##STR00561## ##STR00562##
##STR00563## ##STR00564## ##STR00565## ##STR00566## ##STR00567##
##STR00568## ##STR00569## ##STR00570## ##STR00571## ##STR00572##
##STR00573## ##STR00574## ##STR00575## ##STR00576## ##STR00577##
##STR00578## ##STR00579## ##STR00580## ##STR00581## ##STR00582##
##STR00583## ##STR00584## ##STR00585## ##STR00586## ##STR00587##
##STR00588## ##STR00589## ##STR00590## ##STR00591## ##STR00592##
##STR00593## ##STR00594## ##STR00595## ##STR00596## ##STR00597##
##STR00598## ##STR00599## ##STR00600## ##STR00601## ##STR00602##
##STR00603## ##STR00604## ##STR00605## ##STR00606## ##STR00607##
##STR00608## ##STR00609## ##STR00610## ##STR00611## ##STR00612##
##STR00613## ##STR00614## ##STR00615## ##STR00616## ##STR00617##
##STR00618## ##STR00619## ##STR00620## ##STR00621## ##STR00622##
##STR00623## ##STR00624## ##STR00625## ##STR00626##
##STR00627##
##STR00628## ##STR00629## ##STR00630## ##STR00631## ##STR00632##
##STR00633## ##STR00634## ##STR00635## ##STR00636## ##STR00637##
##STR00638## ##STR00639## ##STR00640## ##STR00641## ##STR00642##
##STR00643## ##STR00644## ##STR00645## ##STR00646## ##STR00647##
##STR00648## ##STR00649## ##STR00650## ##STR00651## ##STR00652##
##STR00653## ##STR00654##
2. Pharmaceutical Formulations.
[0070] The active compounds disclosed herein can, as noted above,
be prepared in the form of their pharmaceutically acceptable salts.
Pharmaceutically acceptable salts are salts that retain the desired
biological activity of the parent compound and do not impart
undesired toxicological effects. Examples of such salts are (a)
acid addition salts formed with inorganic acids, for example
hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric
acid, nitric acid and the like; and salts formed with organic acids
such as, for example, acetic acid, oxalic acid, tartaric acid,
succinic acid, maleic acid, fumaric acid, gluconic acid, citric
acid, malic acid, ascorbic acid, benzoic acid, tannic acid,
palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic
acid, methanesulfonic acid, p-toluenesulfonic acid,
naphthalenedisulfonic acid, polygalacturonic acid, and the like;
(b) salts formed from elemental anions such as chlorine, bromine,
and iodine, and (c) salts derived from bases, such as ammonium
salts, alkali metal salts such as those of sodium and potassium,
alkaline earth metal salts such as those of calcium and magnesium,
and salts with organic bases such as dicyclohexylamine and
N-methyl-D-glucamine.
[0071] Active compounds of the present invention may be prepared as
pharmaceutically acceptable prodrugs. Such prodrugs are those which
are, within the scope of sound medical judgment, suitable for use
in contact with the tissues of humans and lower animals without
undue toxicity, irritation, allergic response and the like,
commensurate with a reasonable risk/benefit ratio, and effective
for their intended use, as well as the zwitterionic forms, where
possible, of the compounds of the invention. The term "prodrug"
refers to compounds that are rapidly transformed in vivo to yield
the parent compound of the above formulae, for example, by
hydrolysis in blood. A thorough discussion is provided in T.
Higuchi and V. Stella, Prodrugs as Novel delivery Systems, Vol. 14
of the A.C.S. Symposium Series and in Edward B. Roche, ed.,
Bioreversible Carriers in Drug Design, American Pharmaceutical
Association and Pergamon Press, 1987, both of which are
incorporated by reference herein. See also U.S. Pat. No. 6,680,299.
Examples include a prodrug that is metabolized in vivo by a subject
to an active drug having an activity of active compounds as
described herein, wherein the prodrug is an ester of an alcohol or
carboxylic acid group, if such a group is present in the compound;
an acetal or ketal of an alcohol group, if such a group is present
in the compound; an N-Mannich base or an imine of an amine group,
if such a group is present in the compound; or a Schiff base,
oxime, acetal, enol ester, oxazolidine, or thiazolidine of a
carbonyl group, if such a group is present in the compound, such as
described in U.S. Pat. No. 6,680,324 and U.S. Pat. No.
6,680,322.
[0072] The active compounds described above may be formulated for
administration in a pharmaceutical carrier in accordance with known
techniques. See, e.g., Remington, The Science and Practice of
Pharmacy (9.sup.th Ed. 1995). In the manufacture of a
pharmaceutical formulation according to the invention, the active
compound (including the physiologically acceptable salts thereof)
is typically admixed with, inter alia, an acceptable carrier. The
carrier must, of course, be acceptable in the sense of being
compatible with any other ingredients in the formulation and must
not be deleterious to the patient. The carrier may be a solid or a
liquid, or both, and is preferably formulated with the compound as
a unit-dose formulation, for example, a tablet, which may contain
from 0.01 or 0.5% to 95% or 99% by weight of the active compound.
One or more active compounds may be incorporated in the
formulations of the invention, which may be prepared by any of the
well known techniques of pharmacy comprising admixing the
components, optionally including one or more accessory
ingredients.
[0073] The formulations of the invention include those suitable for
oral, rectal, topical, buccal (e.g., sub-lingual), vaginal,
parenteral (e.g., subcutaneous, intramuscular, intradermal, or
intravenous), topical (i.e., both skin and mucosal surfaces,
including airway surfaces) and transdermal administration, although
the most suitable route in any given case will depend on the nature
and severity of the condition being treated and on the nature of
the particular active compound which is being used.
[0074] Formulations suitable for oral administration may be
presented in discrete units, such as capsules, cachets, lozenges,
or tablets, each containing a predetermined amount of the active
compound; as a powder or granules; as a solution or a suspension in
an aqueous or non-aqueous liquid; or as an oil-in-water or
water-in-oil emulsion. Such formulations may be prepared by any
suitable method of pharmacy which includes the step of bringing
into association the active compound and a suitable carrier (which
may contain one or more accessory ingredients as noted above). In
general, the formulations of the invention are prepared by
uniformly and intimately admixing the active compound with a liquid
or finely divided solid carrier, or both, and then, if necessary,
shaping the resulting mixture. For example, a tablet may be
prepared by compressing or molding a powder or granules containing
the active compound, optionally with one or more accessory
ingredients. Compressed tablets may be prepared by compressing, in
a suitable machine, the compound in a free-flowing form, such as a
powder or granules optionally mixed with a binder, lubricant, inert
diluent, and/or surface active/dispersing agent(s). Molded tablets
may be made by molding, in a suitable machine, the powdered
compound moistened with an inert liquid binder.
[0075] Formulations suitable for buccal (sub-lingual)
administration include lozenges comprising the active compound in a
flavoured base, usually sucrose and acacia or tragacanth; and
pastilles comprising the compound in an inert base such as gelatin
and glycerin or sucrose and acacia.
[0076] Formulations of the present invention suitable for
parenteral administration comprise sterile aqueous and non-aqueous
injection solutions of the active compound(s), which preparations
are preferably isotonic with the blood of the intended recipient.
These preparations may contain anti-oxidants, buffers,
bacteriostats and solutes which render the formulation isotonic
with the blood of the intended recipient. Aqueous and non-aqueous
sterile suspensions may include suspending agents and thickening
agents. The formulations may be presented in unit\dose or
multi-dose containers, for example sealed ampoules and vials, and
may be stored in a freeze-dried (lyophilized) condition requiring
only the addition of the sterile liquid carrier, for example,
saline or water-for-injection immediately prior to use.
Extemporaneous injection solutions and suspensions may be prepared
from sterile powders, granules and tablets of the kind previously
described. For example, in one aspect of the present invention,
there is provided an injectable, stable, sterile composition
comprising an active compound(s), or a salt thereof, in a unit
dosage form in a sealed container. The compound or salt is provided
in the form of a lyophilizate which is capable of being
reconstituted with a suitable pharmaceutically acceptable carrier
to form a liquid composition suitable for injection thereof into a
subject. The unit dosage form typically comprises from about 10 mg
to about 10 grams of the compound or salt. When the compound or
salt is substantially water-insoluble, a sufficient amount of
emulsifying agent which is physiologically acceptable may be
employed in sufficient quantity to emulsify the compound or salt in
an aqueous carrier. One such useful emulsifying agent is
phosphatidyl choline.
[0077] Formulations suitable for rectal administration are
preferably presented as unit dose suppositories. These may be
prepared by admixing the active compound with one or more
conventional solid carriers, for example, cocoa butter, and then
shaping the resulting mixture.
[0078] Formulations suitable for topical application to the skin
preferably take the form of an ointment, cream, lotion, paste, gel,
spray, aerosol, or oil. Carriers which may be used include
petroleum jelly, lanoline, polyethylene glycols, alcohols,
transdermal enhancers, and combinations of two or more thereof.
[0079] Formulations suitable for transdermal administration may be
presented as discrete patches adapted to remain in intimate contact
with the epidermis of the recipient for a prolonged period of time.
Formulations suitable for transdermal administration may also be
delivered by iontophoresis (see, for example, Pharmaceutical
Research 3 (6):318 (1986)) and typically take the form of an
optionally buffered aqueous solution of the active compound.
Suitable formulations comprise citrate or bis/tris buffer (pH 6) or
ethanol/water and contain from 0.1 to 0.2M active ingredient.
[0080] Further, the present invention provides liposomal
formulations of the compounds disclosed herein and salts thereof.
The technology for forming liposomal suspensions is well known in
the art. When the compound or salt thereof is an aqueous-soluble
salt, using conventional liposome technology, the same may be
incorporated into lipid vesicles. In such an instance, due to the
water solubility of the compound or salt, the compound or salt will
be substantially entrained within the hydrophilic center or core of
the liposomes. The lipid layer employed may be of any conventional
composition and may either contain cholesterol or may be
cholesterol-free. When the compound or salt of interest is
water-insoluble, again employing conventional liposome formation
technology, the salt may be substantially entrained within the
hydrophobic lipid bilayer which forms the structure of the
liposome. In either instance, the liposomes which are produced may
be reduced in size, as through the use of standard sonication and
homogenization techniques.
[0081] Of course, the liposomal formulations containing the
compounds disclosed herein or salts thereof, may be lyophilized to
produce a lyophilizate which may be reconstituted with a
pharmaceutically acceptable carrier, such as water, to regenerate a
liposomal suspension.
[0082] Other pharmaceutical compositions may be prepared from the
water-insoluble compounds disclosed herein, or salts thereof, such
as aqueous base emulsions. In such an instance, the composition
will contain a sufficient amount of pharmaceutically acceptable
emulsifying agent to emulsify the desired amount of the compound or
salt thereof. Particularly useful emulsifying agents include
phosphatidyl cholines, and lecithin.
[0083] In addition to active compound(s), the pharmaceutical
compositions may contain other additives, such as pH-adjusting
additives. In particular, useful pH-adjusting agents include acids,
such as hydrochloric acid, bases or buffers, such as sodium
lactate, sodium acetate, sodium phosphate, sodium citrate, sodium
borate, or sodium gluconate. Further, the compositions may contain
microbial preservatives. Useful microbial preservatives include
methylparaben, propylparaben, and benzyl alcohol. The microbial
preservative is typically employed when the formulation is placed
in a vial designed for multidose use. Of course, as indicated, the
pharmaceutical compositions of the present invention may be
lyophilized using techniques well known in the art.
Dosage and Routes of Administration.
[0084] As noted above, the present invention provides
pharmaceutical formulations comprising the active compounds
(including the pharmaceutically acceptable salts thereof), in
pharmaceutically acceptable carriers for oral, rectal, topical,
buccal, parenteral, intramuscular, intradermal, or intravenous, and
transdermal administration.
[0085] The therapeutically effective dosage of any specific
compound, the use of which is in the scope of present invention,
will vary somewhat from compound to compound, and patient to
patient, and will depend upon the condition of the patient and the
route of delivery. As a general proposition, a dosage from about
0.1 to about 50 mg/kg will have therapeutic efficacy, with all
weights being calculated based upon the weight of the active
compound, including the cases where a salt is employed. Toxicity
concerns at the higher level may restrict intravenous dosages to a
lower level such as up to about 10 mg/kg, with all weights being
calculated based upon the weight of the active base, including the
cases where a salt is employed. A dosage from about 10 mg/kg to
about 50 mg/kg may be employed for oral administration. Typically,
a dosage from about 0.5 mg/kg to 5 mg/kg may be employed for
intramuscular injection.
[0086] Depending upon the disorder being treated, the compounds
described herein may be administered alone or concurrently with one
or more additional active agent useful for treating the disease or
condition with which the patient is afflicted. Examples of
additional active agents include, but are not limited to, those set
forth in paragraphs 0065 through 0387 of W. Hunter, D. Gravett, et
al., US Patent Application Publication No. 20050181977 (Published
Aug. 18, 2005) (assigned to Angiotech International AG) the
disclosure of which is incorporated by reference herein in its
entirety.
[0087] The present invention is explained in greater detail in the
following non-limiting Examples.
Example 1
5-mercapto-benzo[d]isoxazole-4,7-diones and
5-mercapto-1H-indazole-4,7-diones
[0088] Table 2 below shows structure-activity relationships of
5-mercapto-benzo[d]isoxazole-4,7-diones and
5-mercapto-1H-indazole-4,7-diones synthesized in our laboratory.
These compounds (a) inhibit the recombinant thioesterase domain of
FASN; some show marked ability to (b) inhibit FASN in living cells,
as evidenced by the inhibition of .sup.14C-acetate uptake; some
also show marked ability to (c) inhibit in vitro cancer cell
growth.
##STR00655##
[0089] An overall synthetic approach for
5-mercapto-1H-indazole-4,7-diones and other analogs thereof is
shown in Scheme 1. The upper scheme describes the synthesis of
targets where R.sub.4.dbd.H. Briefly 1,4-benzoquinone is reacted
with desired mercaptan according to the method of Katritzky, A. R.;
Odens, H. H.; et al (Rubber Chemistry and Technology, vol 74 (5),
pp. 915-925 (2001)). The resulting 2-mercapto-1,4-benzoquinone is
then treated with diazomethane prepared in situ and in the presence
of and oxidating agent such as
2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) to yield the
desired indazole. The lower scheme describes the synthesis of
targets where R.sub.4 as described in Markush structures Ia, Ib and
Ic is COOEt. Similarly, 1,4-benzoquinone is reacted with desired
mercaptan according to the method of Katritzky, A. R.; Odens, H.
H.; et al (Rubber Chemistry and Technology, vol 74 (5), pp.
915-925, (2001)). The resulting 2-mercapto-1,4-benzoquinone is then
treated with diazomethane derivative prepared by the method of Watt
(J. Org. Chem. vol 51, pp. 5362-536 (1986)) in the presence of an
oxidizing agent (DDQ) to yield the desired indazole compound.
##STR00656##
[0090] An overall synthetic approach for
5-mercapto-benzo[d]isoxazole-4,7-diones and other analogs thereof
is shown in Scheme 2. Briefly 1,4-benzoquinone is reacted with
desired mercaptan according to the method of Katritzky, A. R.;
Odens, H. H.; et al (Rubber Chemistry and Technology, vol 74 (5),
915-925 (2001)) followed by addition of a nitrile oxide derivative
prepared by the method of Hamadi and Msaddek (Heterocyclic
Communications, Vol 12 (6), pp. 457-462 (2006)).
TABLE-US-00002 % inhibi- cell survival, recombinant thioesterase
tion of MTS assay (IC.sub.50) % Inhibi- approx, approx,
.sup.14C-acetate normal tion (10 .mu.M) IC.sub.50 IC.sub.50 incorp.
cells tumor cells Compound Structure Number TE1 TE2 TE1(.mu.M) TE2
(.mu.M) PC3 cells PC3 DU-145 FS-4 ##STR00657## TPI- 00401- 00-A
40.00 55.88 NA NA 29.1 33 >50 ND ##STR00658## TPI- 00402- 00-A
66.63 95.73 2.35 0.56 90.4 18.75 40 20.9 ##STR00659## TPI- 00403-
00-A 69.35 79.27 3.90 2.42 97 (IC.sub.50 = 6.75 .mu.M) 3.25 15.6
9.15 ##STR00660## TPI- 00405- 00-A 69.52 96.82 2.78 0.41 ND ND ND
ND ##STR00661## TPI- 00406- 00-A 73.64 96.01 NA NA 37.7 ND ND ND
##STR00662## TPI- 00407- 00-A 65.96 94.89 NA NA 37.8 ND ND ND
##STR00663## TPI- 00408- 00-A 67.87 96.18 NA NA 89.9 18.5 ND 22.5
##STR00664## TPI- 00409- 00-A 53.50 92.17 NA NA 0 ND ND ND
##STR00665## TPI- 00410- 00-A 59.61 91.67 NA NA 1.85 ND ND ND
##STR00666## TPI- 00411- 00-A 70.51 97.43 NA NA 0.75 ND ND ND
##STR00667## TPI- 00412- 00-A 81.63 97.98 NA NA 9.95 ND ND ND
##STR00668## TPI- 00413- 00-A 50.23 94.78 NA NA 20.2 ND ND ND
##STR00669## TPI- 00414- 00-A 52.21 86.40 NA NA 92.4 25 ND 33
##STR00670## TPI- 00415- 00-A 79.69 97.46 NA NA 7.05 ND ND ND
##STR00671## TPI- 00416- 00-A 64.31 96.49 NA NA 89.9 25.8 ND 33
##STR00672## TPI- 00417- 00-A 74.81 97.85 NA NA 96.1 13.8 ND 21
##STR00673## TPI- 00418- 00-A 65.54 95.92 NA NA 92.2 31.7 ND 37
##STR00674## TPI- 00419- 00-A 66.19 95.79 NA NA 67.95 32 ND 50
##STR00675## TPI- 00420- 00-A 61.96 95.35 NA NA 68.5 37.5 ND 42
##STR00676## TPI- 00421- 00-A 100.00 100.00 1.02 NA 70 17.9 ND 41
##STR00677## TPI- 00422- 00-A 22.92 86.76 NA NA 0 48 ND >50
##STR00678## TPI- 00423- 00-A 79.75 97.99 NA NA 87.25 20 ND 25
##STR00679## TPI- 00424- 00-A 92.59 100.00 NA NA 0 27 ND 50 Key to
Table 2: Recombinant Thioesterase Inhibition Assay. Briefly,
recombinant FASN-TE (250 nM, 100 mM TRIS pH 7.4, 50 mM NaCl, 1 mM
EDTA) was pre-incubated with inhibitors for 30 min at 37.degree. C.
prior to the addition of fluorogenic substrate,
4-methylumbelliferyl heptanoate (4- MUH, 120 .mu.M) (56); the
reaction was then monitored by following the increase in
fluorescence using a Tecan Safire.sup.2 96-well plate reader using
350/450 excitation/emission wavelengths. The IC.sub.50 values are
then determined for each compound in the series. Inhibition of
.sup.14C-Acetate Incorporation in PC3 Cells. This test ensures that
the compound(s) target intracellular FASN correctly. The ability to
inhibit FASN activity in cells will be performed by metabolic
labeling of newly synthesized fatty acids with .sup.14C-acetate, as
we have described previously. Briefly, PC-3 cells were seeded in
24-well plates at 50,000 cells per well. After 48 hours media was
replaced with fresh media containing either drug or DMSO vehicle.
As with the toxicity assay described below, the IC.sub.50 for the
ability to inhibit FASN will also be determined. Cells will be
incubated with a dose of each derivative compound that reflects the
IC.sub.50's determined in the MTS assays. After two hours, 1 .mu.Ci
of .sup.14C-acetate was added to each well for a two hour labeling
period. The cells are then washed with PBS and collected by
trypsinization. Lipids were finally isolated by Folch extraction
and quantified by scintillation counting. The IC.sub.50 value for
each compound against FASN in cells can then be determined. MTS
Cell Survival Assay. Prostate tumor (PC3, DU145) and normal
fibroblast controls (FS-4) cells were individually seeded in
96-well plates at a density of 4,000 cells per well. After 48 hours
the cells are be treated with a dose of each compound (1-50 .mu.M)
or DMSO vehicle in fresh media. Cell viability of each cell line
was then determined after 24, 48 and 72 hours of treatment with
each compound by MTS assay (Roche). MTS assays provide a relative
measure of cell viability based on the ability of mitochondria to
metabolize a tetrazolium substrate to a colored formazan product.
Viability is determined by colorimetric readout at 492 nm and
determined relative to vehicle only treated cells. The IC.sub.50
for each drug, at each time point, will be determined by curve
fitting.
[0091] The foregoing is illustrative of the present invention, and
is not to be construed as limiting thereof. The invention is
defined by the following claims, with equivalents of the claims to
be included therein.
* * * * *