U.S. patent application number 10/847108 was filed with the patent office on 2005-01-20 for branched chain amino acid-dependent aminotransferase inhibitors and their use in the treatment of neurodegenerative diseases.
Invention is credited to Hu, Lain-Yen, Kesten, Suzanne Ross, Lei, Huangshu, Ryder, Todd Robert, Wustrow, David Juergen.
Application Number | 20050014841 10/847108 |
Document ID | / |
Family ID | 23303625 |
Filed Date | 2005-01-20 |
United States Patent
Application |
20050014841 |
Kind Code |
A1 |
Hu, Lain-Yen ; et
al. |
January 20, 2005 |
Branched chain amino acid-dependent aminotransferase inhibitors and
their use in the treatment of neurodegenerative diseases
Abstract
The invention relates to BCAT inhibitors and the use thereof for
treating or preventing neuronal loss associated with stroke,
ischemia, CNS trauma, hypoglycemia and surgery, as well as treating
neurodegenerative diseases including Alzheimer's disease,
amyotrophic lateral sclerosis, Huntington's disease and Down's
syndrome, treating or preventing the adverse consequences of the
overstimulation of the excitatory amino acids, treating anxiety,
psychosis, convulsions, aminoglycoside antibiotics-induced hearing
loss, migraine headache, chronic pain, neuropathic pain,
Parkinson's disease, diabetic retinopathy, glaucoma, CMV retinitis,
urinary incontinence, opioid tolerance or withdrawal, and inducing
anesthesia, as well as for enhancing cognition.
Inventors: |
Hu, Lain-Yen; (Ann Arbor,
MI) ; Kesten, Suzanne Ross; (Ann Arbor, MI) ;
Lei, Huangshu; (Ann Arbor, MI) ; Ryder, Todd
Robert; (Rochester, NY) ; Wustrow, David Juergen;
(Ann Arbor, MI) |
Correspondence
Address: |
SCULLY SCOTT MURPHY & PRESSER, PC
400 GARDEN CITY PLAZA
GARDEN CITY
NY
11530
|
Family ID: |
23303625 |
Appl. No.: |
10/847108 |
Filed: |
May 17, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10847108 |
May 17, 2004 |
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10304803 |
Nov 26, 2002 |
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6809119 |
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60333636 |
Nov 27, 2001 |
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Current U.S.
Class: |
514/602 ;
564/81 |
Current CPC
Class: |
A61P 25/36 20180101;
A61P 9/10 20180101; A61P 27/02 20180101; A61P 25/00 20180101; A61P
25/16 20180101; A61P 25/22 20180101; A61P 13/02 20180101; C07C
311/49 20130101; A61P 43/00 20180101; A61P 3/10 20180101; A61P
25/04 20180101; A61P 25/08 20180101; A61P 25/06 20180101; A61P
25/18 20180101; A61P 21/00 20180101; A61P 25/28 20180101; A61P 3/00
20180101; C07C 2601/14 20170501; A61P 27/06 20180101 |
Class at
Publication: |
514/602 ;
564/081 |
International
Class: |
C07C 311/30 |
Claims
1. A compound of the Formula I 41wherein: R.sub.3 is H, halogen,
alkyl, carboxy, alkoxy, or substituted alkoxy; R.sub.1, R.sub.2,
R.sub.4, and R.sub.5 are independently, H, alkyl, substituted
alkyl, alkoxy, substituted alkoxy, NO.sub.2, halogen, or CF.sub.3;
R.sub.6 or R.sub.7 is independently H, halogen, alkyl, substituted
alkyl, arylalkyl, substituted arylalkyl,
C(CH.sub.2).sub.n--NHSO.sub.2-aryl,
C(CH.sub.2).sub.n--NHSO.sub.2-substituted aryl,
(CH.sub.2).sub.n--NHSO.su- b.2-alkyl,
(CH.sub.2).sub.n--NHSO.sub.2-substituted alkyl,
(CH.sub.2).sub.n--NHSO.sub.2-arylalkyl,
(CH.sub.2).sub.n--NHSO.sub.2-subs- tituted arylalkyl,
NHSO.sub.2-aryl, (CH.sub.2).sub.naryl, (CH.sub.2).sub.n-substituted
aryl, (CH.sub.2).sub.n-alkyl, (CH.sub.2).sub.n-substituted alkyl,
O-alkyl, O-substituted alkyl, (CH.sub.2).sub.n--SO.sub.2NH-aryl,
(CH.sub.2).sub.n--SO.sub.2NH-substitut- ed aryl,
(CH.sub.2).sub.n--SO.sub.2NH-alkyl, (CH.sub.2).sub.n--SO.sub.2NH--
substituted alkyl, (CH.sub.2).sub.n--C(O)NH-arylalkyl,
(CH.sub.2).sub.n--C(O)NH-substituted arylalkyl,
(CH.sub.2).sub.n--C(O)NH-- aryl,
(CH.sub.2).sub.n--C(O)NH-substituted aryl,
(CH.sub.2).sub.n--C(O)NH-- alkyl,
(CH.sub.2).sub.n--C(O)NH-substituted alkyl,
(CH.sub.2).sub.n--SO.su- b.2NH-arylalkyl,
(CH.sub.2).sub.n--SO.sub.2NH-substituted arylalkyl,
(CH.sub.2).sub.n--NH.sub.2, (CH.sub.2).sub.nNH-aryl,
(CH.sub.2).sub.n--NH-substituted aryl, (CH.sub.2).sub.n--NH-alkyl,
(CH.sub.2).sub.n--NH-substituted alkyl,
(CH.sub.2).sub.n--NH-arylalkyl, (CH.sub.2).sub.n--NH-substituted
arylalkyl, (CH.sub.2).sub.n--NHSO.sub.2-- aryl,
(CH.sub.2).sub.n--NHSO.sub.2-substituted aryl,
(CH.sub.2).sub.n--NHC(O)-arylalkyl,
(CH.sub.2).sub.n--NHC(O)-substituted arylalkyl, (C.ident.C)-alkyl,
(C.ident.C)-substituted alkyl, (C.ident.C)-arylalkyl,
(C.ident.C)-substituted arylalkyl, (C.ident.C)-aryl, or
(C.ident.C)-substituted aryl; and n is 0, 1, 2, or 3. or a
pharmaceutically acceptable salt, ester, prodrug, or amide
thereof.
2. A compound according to claim 1 where R.sub.1, R.sub.2, R.sub.3,
R.sub.4 and R.sub.5 are hydrogen.
3. A compound according to claim 1 wherein R.sub.6 or R.sub.7
comprises --NHSO.sub.2-aryl, --NHSO.sub.2-substituted aryl,
--NHSO.sub.2-alkyl or --NHSO.sub.2-substituted alkyl, and R.sub.1,
R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are H.
4. (Cancelled).
5. A compound according to claim 1 wherein the compound is: Benzoic
acid, 4-amino-2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[(2,4,6-trichlorophenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[[4-(trifluoromethyl)phenyl]sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[(3,4-dimethoxyphenyl)sulfo- nyl]amino]-,
2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[[(4-phenoxy)benzene]sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[(2-chloro-6-methylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[(methylsulfonyl)amino]-, 2-(phenylsulfonyl)hydrazide; Benzoic
acid, 4-[[(4-methylphenyl)sulfonyl]a- mino]-,
2-(phenylsulfonyl)hydrazide; Benzoic acid, 4-[[(4-ethylphenyl)sulf-
onyl]amino]-, 2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[(4-n-propylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[(4-n-butylphenyl)sulfonyl]amino]-, 2-(phenylsulfonyl)hydrazide;
Benzoic acid, 4-[[(4-n-pentylphenyl)sulfonyl- ]amino]-,
2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[(4-n-fluorophenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[phenylsulfonyl]amino]-, 2-(phenylsulfonyl)hydrazide;
4-Benzylamino-benzoic acid 2-(phenylsulfonyl)hydrazide; Benzoic
acid, 4-[[(3-methylphenyl)sulfonyl]amino-,
2-(phenylsulfonyl)hydrazide; 4-(1,1-Dimethylethyl)-benzoic acid
2-(phenylsulfonyl)hydrazide; [1,1'-Biphenyl]-4-carboxylic acid
2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[(4-methoxyphenyl)sulfonyl]amino]-, 2-(phenylsulfonyl)hydrazide;
4-Iodo-benzoic acid, 2-(phenylsulfonyl)hydrazide;
4-Phenacetylenyl-benzoi- c acid, 2-(phenylsulfonyl)hydrazide;
Phenethyl-benzoic acid, 2-(phenylsulfonyl)hydrazide;
4-(3-Cyclohexyl-1-propynyl)-benzoic acid,
2-(phenylsulfonyl)hydrazide;
4-(3-Cyclohexyl-3-hydroxy-1-propynyl)-benzoi- c acid,
2-(phenylsulfonyl)hydrazide; 4-(3-Cyclohexyl-3-hydroxy-1-propyl)-b-
enzoic acid, 2-(phenylsulfonyl)hydrazide;
4-(3,3-Dimethyl-1-butynyl)-benzo- ic acid,
2-(phenylsulfonyl)hydrazide; and 4-(3,3-Dimethyl-1-butyl)-benzoic
acid, 2-(phenylsulfonyl)hydrazide.
6. A pharmaceutical composition comprising a compound of claim 1
and pharmaceutically acceptable carrier.
7. A method of treating or preventing neuronal loss associated with
stroke, ischemia, CNS trauma, hypoglycemia or surgery, or treating
a neurodegenerative disease, or treating or preventing the adverse
consequences of the overstimulation of the excitatory amino acids,
or treating anxiety, psychosis, glaucoma, CMV retinitis, diabetic
retinopathy, urinary incontinence, migraine headache, convulsions,
aminoglycoside antibiotics-induced hearing loss, Parkinson's
disease, chronic pain, neuropathic pain, or inducing anesthesia,
opioid tolerance or withdrawal, or enhancing cognition, comprising
administering to a patient in need of such treatment an effective
amount of a compound of claim 1.
8. The method of claim 7 wherein R.sub.6 or R.sub.7 comprises
--NHSO.sub.2-aryl, --NHSO.sub.2-substitued aryl, --NHSO.sub.2-alkyl
or --NHSO.sub.2-substituted alkyl, and R.sub.1, R.sub.2, R.sub.3,
R.sub.4, and R.sub.5 are H.
9. The method of claim 7 wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, and R.sub.5 are hydrogen.
10. (Cancelled).
11. The method of claim 7 wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, and R.sub.5 are hydrogen.
12. The method of claim 7 wherein the compound is: Benzoic acid,
4-amino-2-(phenylsulfonyl)hydrazide.
13. A method for inhibiting branched chain amino acid-dependent
aminotransferase in a patient, the method comprising the step of
administering to the patient a therapeutically effective amount of
a compound of claim 1.
14. The method of claim 13, wherein R.sub.6 or R.sub.7 comprises
--NHSO.sub.2-aryl, --NHSO.sub.2-substitued aryl, --NHSO.sub.2-alkyl
or --NHSO.sub.2-substituted alkyl, and R.sub.1, R.sub.2, R.sub.3,
R.sub.4, and R.sub.5 are H.
15. (Cancelled).
16. The method of claim 13, wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, and R.sub.5 are hydrogen.
17. The method of claim 13, wherein the compound is: Benzoic acid,
4-amino-2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[[4-(trifluoromethyl)phenyl]sulfonyl]amino]-,
2-(phenylsulfonyl)hydraz- ide; Benzoic acid,
4-[[(3,4-dimethoxyphenyl)sulfonyl]amino]-2-(phenylsulfo-
nyl)hydrazide; Benzoic acid,
4-[[[(4-phenoxy)benzene]sulfonyl]amino]-2-(ph-
enylsulfonyl)hydrazide; Benzoic acid,
4-[[(2-chloro-6-methylphenyl)sulfony- l]amino]-,
2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[(methylsulfonyl)amino]-, 2-(phenylsulfonyl)hydrazide; Benzoic
acid,
4-[[(4-methylphenyl)sulfonyl]amino]-2-(phenylsulfonyl)hydrazide;
Benzoic acid,
4-[[(4-ethylphenyl)sulfonyl]amino]-2-(phenylsulfonyl)hydrazide;
Benzoic acid,
4-[[(4-n-propylphenyl)sulfonyl]amino]-2-(phenylsulfonyl)hyd-
razide; Benzoic acid,
4-[[(4-n-butylphenyl)sulfonyl]amino]-2-(phenylsulfon- yl)hydrazide;
Benzoic acid, 4-[[(4-n-pentylphenyl)sulfonyl]amino]-2-(pheny-
lsulfonyl)hydrazide; Benzoic acid,
4-[[(4-fluorophenyl)sulfonyl]amino]-2-(- phenylsulfonyl)hydrazide;
Benzoic acid, 4-[phenylsulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide; 4-Benzylamino-benzoic acid
2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[(3-methylphenyl)sulfonyl]a- mino],
-2-(phenylsulfonyl)hydrazide; 4-Phenoxybenzoic acid,
2-[(3-methyl)phenylsulfonyl]hydrazide; 4-Phenoxybenzoic acid,
2-[3-bromophenylsulfonyl]hydrazide; 4-Phenoxybenzoic acid,
2-[2-chlorophenylsulfonyl]hydrazide; 4-Phenoxybenzoic acid,
2-[2-(trifluoromethoxy)phenylsulfonyl]hydrazide; 4-Phenoxybenzoic
acid, 2-[(4-methyl-3-nitro)phenylsulfonyl]hydrazide;
4-Phenoxybenzoic acid, 2-[2-trifluorophenylsulfonyl]hydrazide;
4-(1,1-Dimethylethyl)-benzoic acid 2-(phenylsulfonyl)hydrazide;
[1,1'-Biphenyl]-4-carboxylic acid 2-(phenylsulfonyl)hydrazide;
3-Phenoxybenzoic acid, 2-(phenylsulfonyl)hydrazide;
2-Phenoxybenzoic acid, 2-(phenylsulfonyl)hydrazide;
4-Phenoxybenzoic acid, 2-(phenylsulfonyl)hydrazide;
4-Benzyloxybenzoic acid, 2-(phenylsulfonyl)hydrazide; Benzoic acid,
4-[[(4-methoxyphenyl)sulfonyl]- amino]-,
2-(phenylsulfonyl)hydrazide; 4-Iodo-benzoic acid,
2-(phenylsulfonyl)hydrazide; 4-Phenacetylenyl-benzoic acid,
2-(phenylsulfonyl)hydrazide; Phenacetylenyl-benzoic acid,
2-(phenylsulfonyl)hydrazide; 4-(3-Cyclohexyl-1-propynyl)-benzoic
acid, 2-(phenylsulfonyl)hydrazide;
4-(3-Cyclohexyl-3-hydroxy-1-propynyl)-benzoi- c acid,
2-(phenylsulfonyl)hydrazide; 4-(3-Cyclohexyl-3-hydroxy-1-propyl)-b-
enzoic acid, 2-(phenylsulfonyl)hydrazide;
4-(3,3-Dimethyl-1-butynyl)-benzo- ic acid,
2-(phenylsulfonyl)hydrazide; and 4-(3,3-Dimethyl-1-butyl)-benzoic
acid, 2-(phenylsulfonyl)hydrazide.
Description
[0001] This application claims benefit of U.S. Provisional
Application No. 60/333,636 filed Nov. 27, 2001.
FIELD OF THE INVENTION
[0002] This invention is related to branched chain amino
acid-dependent amino transferase (BCAT) inhibitors. The invention
is also directed to the use of BCAT inhibitors as neuro-protective
agents for treating conditions such as stroke, cerebral ischemia,
central nervous system trauma, hypoglycemia, anxiety, convulsions,
aminoglycoside antibiotics-induced hearing loss, migraine
headaches, chronic pain, neuropathic pain, glaucoma, CMV retinitis,
diabetic retinopathy, psychosis, urinary incontinence, opioid
tolerance or withdrawal, or neuro-degenerative disorders such as
lathyrism, Alzheimer's disease, Parkinsonism, amyotrophic lateral
sclerosis (ALS), and Huntington's Disease.
RELATED BACKGROUND ART
[0003] Excessive excitation by neurotransmitters can cause the
degeneration and death of neurons. It is believed that this
degeneration is in part mediated by the excitotoxic actions of the
excitatory amino acids (EAA) glutamate and aspartate at the
N-methyl-D-aspartate (NMDA) receptor. This excitotoxic action is
considered responsible for the loss of neurons in cerebrovascular
disorders such as cerebral ischemia or cerebral infarction
resulting from a range of conditions, such as thromboembolic or
hemorrhagic stroke, cerebral vasospasms, hypoglycemia, cardiac
arrest, status epilepticus, perinatal asphyxia, anoxia such as from
drowning, pulmonary surgery and cerebral trauma, as well as
lathyrism, Alzheimer's disease, Parkinson's disease, and
Huntington's disease.
[0004] Excitatory amino acid receptor antagonists that block NMDA
receptors are recognized for usefulness in the treatment of
disorders. NMDA receptors are intimately involved in the phenomenon
of excitotoxicity, which may be a critical determinant of outcome
of several neurological disorders. Disorders known to be responsive
to blockade of the NMDA receptor include acute cerebral ischemia
(stroke or cerebral trauma, for example), muscular spasm,
convulsive disorders, neuropathic pain and anxiety, and may be a
significant causal factor in chronic neurodegenerative disorders
such as Parkinson's disease (Klockgether T., Turski L., Ann.
Neurol., 1993;34:585-593), human immunodeficiency virus (HIV)
related neuronal injury, amyotrophic lateral sclerosis (ALS),
Alzheimer's disease (Francis P. T., Sims N. R., Procter A. W.,
Bowen D. M., J. Neurochem., 1993;60(5):1589-1604, and Huntington's
disease (see Lipton S., TINS, 1993;16(12):527-532; Lipton S. A.,
Rosenberg P. A., New Eng. J. Med., 1994;330(9):613-622; and Bigge
C. F., Biochem. Pharmacol., 1993;45:1547-1561, and referenced cited
therein). NMDA receptor antagonists may also be used to prevent
tolerance to opiate analgesia or to help control withdrawal
symptoms from addictive drugs (Eur. Pat. Appl. 488,959A).
[0005] U.S. Pat. No. 5,352,683 discloses the treatment of chronic
pain with a compound which is an antagonist of the NMDA
receptor.
[0006] U.S. Pat. No. 4,902,695 discloses certain competitive NMDA
antagonists that are useful for the treatment of neurological
disorders, including epilepsy, stroke, anxiety, cerebral ischemia,
muscular spasms, and neurodegenerative diseases such as Alzheimer's
disease and Huntington's disease.
[0007] U.S. Pat. No. 5,192,751 discloses a method of treating
urinary incontinence in a mammal which comprises administering an
effective amount of a competitive NMDA antagonist.
SUMMARY OF THE INVENTION
[0008] The invention relates BCAT inhibitor compounds of Formula I
1
[0009] wherein:
[0010] R.sub.3 is H, halogen, alkyl, carboxy, alkoxy, substituted
alkoxy;
[0011] R.sub.1, R.sub.2, R.sub.4, and R.sub.5 are independently, H,
alkyl, substituted alkyl, alkoxy, substituted alkoxy, NO.sub.2,
halogen, or CF.sub.3;
[0012] R.sub.6 or R.sub.7 are independently, H, halogen, alkyl,
substituted alkyl, arylalkyl, substituted arylalkyl,
C(CH.sub.2).sub.n--NHSO.sub.2-aryl,
C(CH.sub.2).sub.n--NHSO.sub.2-substit- uted aryl,
(CH.sub.2).sub.n--NHSO.sub.2-alkyl, (CH.sub.2).sub.n--NHSO.sub.-
2-substituted alkyl, (CH.sub.2).sub.n--NHSO.sub.2-arylalkyl,
(CH.sub.2).sub.n--NHSO.sub.2-substituted arylalkyl,
NHSO.sub.2-aryl, NHSO.sub.2-substituted aryl, NHSO.sub.2-alkyl,
NHSO.sub.2-substituted alkyl, NHSO.sub.2-arylalkyl,
NHSO.sub.2-substituted arylalkyl, (CH.sub.2).sub.n-aryl,
(CH.sub.2).sub.n-substituted aryl, (CH.sub.2).sub.n-alkyl,
(CH.sub.2).sub.n-substituted alkyl, O-aryl, O-substituted aryl,
O-alkyl, O-substituted alkyl, O-arylalkyl, O-substituted arylalkyl,
(CH.sub.2).sub.n--SO.sub.2NH-aryl,
(CH.sub.2).sub.n--SO.sub.2NH-substituted aryl,
(CH.sub.2).sub.n--SO.sub.2- NH-alkyl,
(CH.sub.2).sub.n--SO.sub.2NH-substituted alkyl,
(CH.sub.2).sub.n--C(O)NH-arylalkyl,
(CH.sub.2).sub.n--C(O)NH-substituted arylalkyl,
(CH.sub.2).sub.n--C(O)NH-aryl, (CH.sub.2).sub.n--C(O)NH-substi-
tuted aryl, (CH.sub.2).sub.n--C(O)NH-alkyl,
(CH.sub.2).sub.n--C(O)NH-subst- ituted alkyl,
(CH.sub.2).sub.n--SO.sub.2NH-arylalkyl,
(CH.sub.2).sub.n--SO.sub.2NH-substituted arylalkyl,
(CH.sub.2).sub.n--NH.sub.2, (CH.sub.2).sub.n--NH-aryl,
(CH.sub.2).sub.n--NH-substituted aryl, (CH.sub.2).sub.n--NH-alkyl,
(CH.sub.2).sub.n--NH-substituted alkyl,
(CH.sub.2).sub.n--NH-arylalkyl, (CH.sub.2).sub.n--NH-substituted
arylalkyl, (CH.sub.2).sub.n--NHSO.sub.2-- aryl,
(CH.sub.2).sub.n--NHSO.sub.2-substituted aryl,
(CH.sub.2).sub.n--NHSO.sub.2-alkyl,
(CH.sub.2).sub.n--NHSO.sub.2-substitu- ted alkyl,
(CH.sub.2).sub.n--NHC(O)-arylalkyl, (CH.sub.2).sub.n--NHC(O)-su-
bstituted arylalkyl, (CH.sub.2).sub.n--NO.sub.2, (C.ident.C)-alkyl,
(C.ident.C)-substituted alkyl, (C.ident.C)-arylalkyl,
(C.ident.C)-substituted arylalkyl, (C.ident.C)-aryl, or
(C.ident.C)-substituted aryl; and
[0013] n is 0, 1, 2, or 3.
[0014] or a pharmaceutically acceptable salt, ester, prodrug, or
amide thereof;
[0015] where there is more than one stereoisomer, each chiral
center may be independently R or S; or a pharmaceutically
acceptable salt, ester, prodrug, or amide thereof;
[0016] The invention also relates to compounds of Formula I,
wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5 are
hydrogen; and R.sub.6 or R.sub.7 is --NHSO.sub.2-aryl,
--NHSO.sub.2-substituted aryl, --NHSO.sub.2-alkyl, or
--NHSO.sub.2-substituted alkyl.
[0017] The invention also relates to compounds selected from:
[0018] Benzoic acid, 4-nitro-2-(phenylsulfonyl)hydrazide;
[0019] Benzoic acid, 4-amino-2-(phenylsulfonyl)hydrazide;
[0020] Benzoic acid, 4-[[(2,4,6-trichlorophenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0021] Benzoic acid,
4-[[[4-(trifluoromethyl)phenyl]sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0022] Benzoic acid, 4-[[(3,4-dimethoxyphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0023] Benzoic acid, 4-[[[(4-phenoxy)benzene]sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0024] Benzoic acid, 4-[[(2-chloro-6-methylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0025] Benzoic acid, 4-[(methylsulfonyl)amino]-,
2-(phenylsulfonyl)hydrazi- de;
[0026] Benzoic acid, 4-[[(4-methylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0027] Benzoic acid, 4-[[(4-ethylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0028] Benzoic acid, 4-[[(4-n-propylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0029] Benzoic acid, 4-[[(4-n-butylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0030] Benzoic acid, 4-[[(4-n-pentylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0031] Benzoic acid, 4-[[(4-fluorophenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0032] Benzoic acid, 4-[phenylsulfonyl]amino]-,
2-(phenylsulfonyl)hydrazid- e;
[0033] 4-Benzylamino-benzoic acid 2-(phenylsulfonyl)hydrazide;
[0034] Benzoic acid, 4-[[(3-methylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0035] 4-Phenoxybenzoic acid,
2-[(3-methyl)phenylsulfonyl]hydrazide;
[0036] 4-Phenoxybenzoic acid,
2-[3-bromophenylsulfonyl]hydrazide;
[0037] 4-Phenoxybenzoic acid,
2-[2-chlorophenylsulfonyl]hydrazide;
[0038] 4-Phenoxybenzoic acid,
2-[2-(trifluoromethoxy)phenylsulfonyl]hydraz- ide;
[0039] 4-Phenoxybenzoic acid,
2-[(4-methyl-3-nitro)phenylsulfonyl]hydrazid- e;
[0040] 4-Phenoxybenzoic acid,
2-[2-trifluorophenylsulfonyl]hydrazide;
[0041] 4-(1,1-Dimethylethyl)-benzoic acid
2-(phenylsulfonyl)hydrazide;
[0042] [1,1'-Biphenyl]4-carboxylic acid
2-(phenylsulfonyl)hydrazide;
[0043] 3-Phenoxybenzoic acid, 2-(phenylsulfonyl)hydrazide;
[0044] 2-Phenoxybenzoic acid, 2-(phenylsulfonyl)hydrazide;
[0045] 4-Phenoxybenzoic acid, 2-(phenylsulfonyl)hydrazide;
[0046] 4-Benzyloxybenzoic acid, 2-(phenylsulfonyl)hydrazide;
[0047] Benzoic acid, 4-[[(4-methoxyphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide;
[0048] 4-Iodo-benzoic acid, 2-(phenylsulfonyl)hydrazide;
[0049] 4-Phenacetylenyl-benzoic acid,
2-(phenylsulfonyl)hydrazide;
[0050] Phenethyl-benzoic acid, 2-(phenylsulfonyl)hydrazide;
[0051] 4-(3-Cyclohexyl-1-propynyl)-benzoic acid,
2-(phenylsulfonyl)hydrazi- de;
[0052] 4-(3-Cyclohexyl-3-hydroxy-1-propynyl)-benzoic acid,
2-(phenylsulfonyl)hydrazide;
[0053] 4-(3-Cyclohexyl-3-hydroxy-1-propyl)-benzoic acid,
2-(phenylsulfonyl)hydrazide;
[0054] 4-(3,3-Dimethyl-1-butynyl)-benzoic acid,
2-(phenylsulfonyl)hydrazid- e; and
[0055] 4-(3,3-Dimethyl-1-butyl)-benzoic acid,
2-(phenylsulfonyl)hydrazide.
[0056] The invention also relates to a method of treating or
preventing neuronal loss associated with stroke, ischemia, CNS
trauma, hypoglycemia and surgery, as well as treating
neurodegenerative diseases including Alzheimer's disease,
amyotrophic lateral sclerosis, Huntington's disease, Parkinson's
disease and Down's syndrome, treating or preventing the adverse
consequences of the overstimulation of the excitatory amino acids,
treating anxiety, psychosis, convulsions, chronic pain, neuropathic
pain, diabetic retinopathy, glaucoma, CMV retinitis, urinary
incontinence, and inducing anesthesia, as well as enhancing
cognition, and preventing opiate tolerance and withdrawal symptoms,
comprising administering to an animal in need of such treatment an
effective amount of any one of the BCAT inhibitors of the present
invention, or a pharmaceutically acceptable salt thereof.
[0057] The present invention also includes a pharmaceutical
composition comprising a therapeutically effective amount of one or
more compounds of Formula I and a pharmaceutically acceptable
carrier.
DETAILED DESCRIPTION OF THE INVENTION
[0058] The present invention is predicated on the discovery that
intraocular glutamate injection causes ganglion cell loss which is
related to the early stages of diabetic retinopathy (Vorwerk et
al., IOVS, 1996;37:1618-1624) and that an inhibitor of the branched
chain amino acid-dependent aminotransferase pathway, specifically
gabapentin, is effective in inhibiting the synthesis of glutamate
(see example below), thus preventing diabetic retinopathy.
Accordingly, the present invention provides a method for the
prophylactic and therapeutic treatment of diabetic retinopathy,
including treatment at the pre-diabetic retinopathy stage, the
nonproliferative diabetic retinopathy stage, and the proliferative
diabetic retinopathy stage. By "prophylactic" is meant the
protection, in whole or in part, against diabetic retinopathy, in
particular diabetic macular edema. By "therapeutic" is meant the
amelioration of diabetic retinopathy, itself, and the protection,
in whole or in part, against further diabetic retinopathy, in
particular diabetic macular edema.
[0059] The method comprises the administration of an inhibitor of
the branched chain amino acid-dependent aminotransferase pathway in
an amount sufficient to treat the neurodegenerative disease or
condition, for example, to treat the retina for retinopathy
prophylactically or therapeutically. Any inhibitor of the branched
chain amino acid-dependent aminotransferase pathway can be used in
the method of the present invention as long as it is safe and
efficacious. Herein, "branch chain amino acid-dependent
aminotransferase (BCAT) inhibitor" will be used to refer to such
compounds and is intended to encompass all compounds that affect
the branch chain amino acid-dependent aminotransferase pathway at
any and all points in the pathway.
[0060] Preferably, the BCAT inhibitor is a compound of Formula I as
described above, or a pharmaceutically acceptable, BCAT
pathway-inhibiting analogue or prodrug thereof or a
pharmaceutically acceptable salt, ester, or amide of any of the
foregoing.
[0061] Unless otherwise expressly stated, the following definitions
are adhered to throughout this disclosure.
[0062] The term "alkyl" includes saturated aliphatic groups,
including straight-chain alkyl groups (e.g., methyl, ethyl, propyl,
butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc.),
branched-chain alkyl groups (isopropyl, tert-butyl, isobutyl, etc),
cycloalkyl (alicyclic) groups (e.g., cyclopropyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl), alkyl substituted cycloalkyl
groups, and cycloalkyl substituted alkyl groups. The term "alkyl"
further includes alkyl groups, which can further include oxygen,
nitrogen, sulfur, or phosphorous atoms replacing one or more
carbons of the hydrocarbon backbone. These heteroatoms can also be
substituted. For example, sulfur may be substituted with one or
more oxygen atoms (e.g., SO or SO.sub.2). In certain embodiments, a
straight-chain or branched-chain alkyl has 6 or fewer carbon atoms
in its backbone (e.g., C.sub.1-C.sub.6 for straight chain,
C.sub.3-C.sub.6 for branched chain), and more preferably 4 or
fewer. Likewise, preferred cycloalkyls have from 3 to 8 carbon
atoms in their ring structure, and more preferably have 5 or 6
carbons in the ring structure. The term C.sub.1-C.sub.6 includes
alkyl groups containing 1 to 6 carbon atoms.
[0063] Moreover, the term "alkyl" includes both "unsubstituted
alkyls" and "substituted alkyls," the latter of which refers to
alkyl moieties having substituents replacing a hydrogen on one or
more carbons of the hydrocarbon backbone.
[0064] The term "alkyl" also includes the side chains of natural
and unnatural amino acids. In an embodiment, the term "cycloalkyl"
includes saturated hydrocarbon rings having 3 to 8 carbon atoms and
further includes, for example, cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl, cycloocytl, and the like.
[0065] Unless the number of carbons is otherwise specified, "lower
alkyl" as used herein means an alkyl group, as defined above, but
having from 1 to 5 carbon atoms in its backbone structure. In an
embodiment, the term "lower alkyl" includes straight or branched
hydrocarbon radicals having from 1 to 4 carbon atoms and includes,
for example, methyl, ethyl, n-propyl, isopropyl, n-butyl,
sec-butyl, isobutyl, or tert-butyl. "Lower alkenyl" and "lower
alkynyl" have chain lengths of, for example, 2 to 5 carbon
atoms.
[0066] The term "substituted" includes substitution by one or more
substituents selected from, for example, alkyl, alkenyl, alkynyl,
halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxy, aryloxy,
phosphate, phosphonato, phosphinato, cyano, amino (including alkyl
amino, dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl, and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfate, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.
Cycloalkyls can be further substituted, e.g., with the substituents
described above. An "alkylaryl" or an "arylalkyl" moiety is an
alkyl substituted with an aryl [e.g., phenylmethyl (benzyl)].
[0067] The term "aryl" includes groups, including 5- and 6-membered
single-ring aromatic groups that may include from 0 to 4
heteroatoms, for example, benzene, phenyl, pyrrole, furan,
thiophene, thiazole, isothiaozole, imidazole, triazole, tetrazole,
pyrazole, oxazole, isooxazole, pyridine, pyrazine, pyridazine, and
pyrimidine, and the like. Furthermore, the term "aryl" includes
multicyclic aryl groups, e.g., tricyclic, bicyclic, e.g.,
naphthalene, benzoxazole, benzodioxazole, benzothiazole,
benzoimidazole, benzothiophene, methylenedioxyphenyl, quinoline,
isoquinoline, napthridine, indole, benzofuran, purine, benzofuran,
deazapurine, or indolizine. Those aryl groups having heteroatoms in
the ring structure may also be referred to as "aryl heterocycles,"
"heterocycles," "heteroaryls," or "heteroaromatics". The aromatic
ring can be substituted at one or more ring positions with such
substituents as described above, as for example, halogen, hydroxyl,
alkoxy, aryloxy, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
alkylaminocarbonyl, arylalkyl aminocarbonyl, alkenylaminocarbonyl,
alkylcarbonyl, arylcarbonyl, arylalkylcarbonyl, alkenylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkylamino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl, and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.
Aryl groups can also be fused or bridged with alicyclic or
heterocyclic rings which are not aromatic so as to form a polycycle
(e.g., tetralin). In an embodiment, the heteroaromatic group is 2-
or 3-thienyl, 2- or 3-furanyl, 2- or 3-pyrrolyl, 2-, 3-, or
4-pyridinyl, 2-pyrazinyl, 2-, 4-, or 5-pyrimidinyl, 3- or
4-pyridazinyl, or 2-, 3-, 4-, 5-, 6-, or 7-indolyl.
[0068] Other examples of heterocycles include 5- or 6-membered
mono- or bicyclic ring structures which may contain one or more
heteroatoms such as N, S, or O; examples of heterocycles are
pyridine, pyrimidine, pyridazine, pyrazole, oxazole, indole,
N-alkylindole, quinoline, quinazoline, quinazolinone, and the like.
Substituents may include substituted or unsubstituted lower alkyl,
substituted or unsubstituted alkenyl, substituted or unsubstituted
alkynyl, substituted or unsubstituted araalkyl, substituted or
unsubstituted aryl, halogen, cyano, nitro, CHO, COR.sub.8,
OR.sub.8, COOR.sub.8, CONR.sub.8R.sub.9,
(CH.sub.2).sub.nNR.sub.8R.sub.9, (CH.sub.2).sub.nOR.sub.8,
SR.sub.8, SOR.sub.8, SO.sub.2R.sub.8, NHSO.sub.2R.sub.8,
NR.sub.8R.sub.9, NHCOR.sub.8, O(CR.sub.8R.sub.9).sub.0-3CF.sub.3,
O(R.sub.8R.sub.9).sub.0-- 3CCl.sub.3, or SO.sub.2NR.sub.8R.sub.9,
SCF.sub.3, SCCl.sub.3, wherein R.sub.8 and R.sub.9 independently
hydrogen, substituted or unsubstituted lower alkyl, substituted or
unsubstituted lower alkenyl, substituted or unsubstituted alkynyl,
substituted or unsubstituted araalkyl.
[0069] The term "phenyl" includes aromatic rings of 6 carbons.
Phenyl may be unsubstituted (or substituted with hydrogen) or
substituted at one or more positions with a substituent such as,
but not limited to, alkyl, alkenyl, alkynyl, halogen, hydroxyl
alkoxy, aryloxy, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
alkylaminocarbonyl, arylalkyl aminocarbonyl, alkenylaminocarbonyl,
alkylcarbonyl, arylcarbonyl, arylalkylcarbonyl, alkenylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl, and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.
The term "aryl" is intended to include both substituted and
unsubstituted phenyl groups.
[0070] The term "acyl" includes compounds and moieties which
contain the acyl radical (CH.sub.3CO--) or a carbonyl group. The
term "substituted acyl" includes acyl groups where one or more of
the hydrogen atoms are replaced by for example, alkyl groups,
alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,
alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato,
cyano, amino (including alkyl amino, dialkylamino, arylamino,
diarylamino, and alkylarylamino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl, and ureido),
amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,
sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moiety.
[0071] The term "acylamino" includes moieties wherein an acyl
moiety is bonded to an amino group. For example, the term includes
alkylcarbonylamino, arylcarbonylamino, carbamoyl, and ureido
groups.
[0072] The term "aroyl" includes compounds and moieties with an
aryl or heteroaromatic moiety bound to a carbonyl group. Examples
of aroyl groups include phenylcarboxy, naphthyl carboxy, etc.
[0073] The terms "alkoxyalkyl," "alkylaminoalkyl," and
"thioalkoxyalkyl" include alkyl groups, as described above, which
further include oxygen, nitrogen, or sulfur atoms replacing one or
more carbons of the hydrocarbon backbone, e.g., oxygen, nitrogen,
or sulfur atoms.
[0074] The term "alkoxy" includes substituted and unsubstituted
alkyl, alkenyl, and alkynyl groups covalently linked to an oxygen
atom. Examples of alkoxy groups include methoxy, ethoxy,
isopropyloxy, propoxy, butoxy, and pentoxy groups. Examples of
substituted alkoxy groups include halogenated alkoxy groups. The
alkoxy groups can be substituted with groups such as alkenyl,
alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkylamino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl, and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moieties.
Examples of halogen substituted alkoxy groups include, but are not
limited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy,
chloromethoxy, dichloromethoxy, trichloromethoxy, etc. In an
embodiment, the term "lower alkoxy" includes O-alkyl, wherein alkyl
is as defined above for lower alkyl.
[0075] The term "ether" includes compounds or moieties which
contain an oxygen bonded to two different carbon atoms or
heteroatoms. For example, the term includes "alkoxyalkyl" which
refers to an alkyl, alkenyl, or alkynyl group covalently bonded to
an oxygen atom which is covalently bonded to another alkyl
group.
[0076] The term "ester" includes compounds and moieties which
contain a carbon or a heteroatom bound to an oxygen atom which is
bonded to the carbon of a carbonyl group. The term "ester" includes
alkoxycarboxy groups such as methoxycarbonyl, ethoxycarbonyl,
propoxycarbonyl, butoxycarbonyl, pentoxycarbonyl, etc. The alkyl,
alkenyl, or alkynyl groups are as defined above.
[0077] The term "thioether" includes compounds and moieties which
contain a sulfur atom bonded to two different carbon or hetero
atoms. Examples of thioethers include, but are not limited to,
alkthioalkyls, alkthioalkenyls, and alkthioalkynyls. The term
"alkthioalkyls" includes compounds with an alkyl, alkenyl, or
alkynyl group bonded to a sulfur atom which is bonded to an alkyl
group. Similarly, the terms "alkthioalkenyls" and "alkthioalkynyls"
refer to compounds or moieties wherein an alkyl, alkenyl, or
alkynyl group is bonded to a sulfur atom which is covalently bonded
to an alkynyl group.
[0078] The term "hydroxy" or "hydroxyl" includes groups with an
--OH or --O--. The term "halogen" includes fluorine, bromine,
chlorine, iodine, etc. The term "perhalogenated" generally refers
to a moiety wherein all hydrogens are replaced by halogen
atoms.
[0079] It will be noted that the structure of some of the compounds
of this invention includes asymmetric carbon atoms. It is to be
understood accordingly that the isomers arising from such asymmetry
(e.g., all enantiomers and diastereomers) are included within the
scope of this invention, unless indicated otherwise. Such isomers
can be obtained in substantially pure form by classical separation
techniques and by stereochemically controlled synthesis.
Furthermore, the structures and other compounds and moieties
discussed in this application also include all tautomers
thereof.
[0080] The term "arenes" means a hydrocarbon including an aryl
group.
[0081] The term "subject" includes organisms which are capable of
having or have a chemokine mediated disorder. Preferred examples
include humans and animals, including cows, sheep, pigs, dogs,
cats, rats, ferrets, bears, rabbits, etc.
[0082] The term "treatment" includes the diminishment or
alleviation of at least one symptom associated or caused by the
disorder being treated. For example, treatment can be diminishment
of several symptoms of a disorder or complete eradication of a
disorder.
[0083] The symbol "--" means a bond.
[0084] The compounds of the present invention can exist in
unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms, including hydrated forms,
are equivalent to unsolvated forms and are intended to be
encompassed within the scope of the present invention.
[0085] Certain of the compounds of the present invention possess
one or more chiral centers and each center may exist in the R(D) or
S(L) configuration. The present invention includes all enantiomeric
and epimeric forms as well as the appropriate mixtures thereof.
[0086] The compounds of Formula I are capable of further forming
both pharmaceutically acceptable formulations comprising salts,
esters, amides, and prodrugs. As used herein, the term
"pharmaceutically acceptable salts, esters, amides, and prodrugs"
refers to those carboxylate salts, amino acid addition salts,
esters, amides, and prodrugs of the compounds of the present
invention which are, within the scope of sound medical judgment,
suitable for use in contact with the tissues of patients without
undue toxicity, irritation, allergic response, and the like,
commensurate with a reasonable benefit/risk ratio, and effective
for their intended use, as well as the zwitterionic forms, where
possible, of the compounds of the invention. The term "salts"
refers to the relatively non-toxic, inorganic and organic acid
addition salts of compounds of the present invention. These salts
can be prepared in situ during the final isolation and purification
of the compounds or by separately reacting the purified compound in
its free base form with a suitable organic or inorganic acid and
isolating the salt thus formed and including, but not limited to,
acid addition and/or base salts, solvents and N-oxides of a
compound of Formula I. This invention also provides pharmaceutical
formulations comprising a compound of Formula I together with a
pharmaceutically acceptable carrier, diluent, or excipient
therefor. All of these forms are within the present invention.
[0087] Pharmaceutically acceptable acid addition salts of the
compounds of Formula I include salts derived form inorganic acids
such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic,
hydriodic, phosphorus, and the like, as well as the salts derived
from organic acids, such as aliphatic mono- and dicarboxylic acids,
phenyl-substituted alkanoic acids, hydroxy alkanoic acids,
alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic
acids, etc. Such salts thus include sulfate, pyrosulfate,
bisulfate, sulfite, bisulfite, nitrate, phosphate,
monohydrogenphosphate, dihydrogenphosphate, metaphosphate,
pyrophosphate, chloride, bromide, iodide, acetate, propionate,
caprylate, isobutyrate, oxalate, malonate, succinate, suberate,
sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate,
methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate,
toluenesulfonate, phenylacetate, citrate, lactate, maleate,
tartrate, methanesulfonate, and the like. Also contemplated are the
salts of amino acids such as arginate, gluconate, galacturonate,
and the like; see, for example, Berge, et al., "Pharmaceutical
Salts," J. of Pharmaceutical Science, 1977;66:1-19.
[0088] The acid addition salts of the basic compounds are prepared
by contacting the free base form with a sufficient amount of the
desired acid to produce the salt in the conventional manner. The
free base form may be regenerated by contacting the salt form with
a base and isolating the free base in the conventional manner. The
free base forms differ from their respective salt forms somewhat in
certain physical properties such as solubility in polar solvents,
but otherwise the salts are equivalent to their respective free
base for purposes of the present invention.
[0089] Pharmaceutically acceptable base addition salts are formed
with metals or amines, such as alkali and alkaline earth metal
hydroxides, or of organic amines. Examples of metals used as
cations are sodium, potassium, magnesium, calcium, and the like.
Examples of suitable amines are N,N'-dibenzylethylenediamine,
chloroprocaine, choline, diethanolamine, ethylenediamine,
N-methylglucamine, and procaine; see, for example, Berge et al.,
supra., 1977 both of which are incorporated herein by
reference.
[0090] The base addition salts of acidic compounds are prepared by
contacting the free acid form with a sufficient amount of the
desired base to produce the salt in the conventional manner. The
free acid form may be regenerated by contacting the salt form with
an acid and isolating the free acid in a conventional manner. The
free acid forms differ from their respective salt forms somewhat in
certain physical properties such as solubility in polar solvents,
but otherwise the salts are equivalent to their respective free
acid for purposes of the present invention.
[0091] Examples of pharmaceutically acceptable, non-toxic esters of
the compounds of this invention include C.sub.1-C.sub.6 alkyl
esters wherein the alkyl group is a straight or branched chain.
Acceptable esters also include C.sub.5-C.sub.7 cycloalkyl esters as
well as arylalkyl esters such as, but not limited to benzyl.
C.sub.1-C.sub.4 alkyl esters are preferred. Esters of the compounds
of the present invention may be prepared according to conventional
methods.
[0092] Examples of pharmaceutically acceptable, non-toxic amides of
the compounds of this invention include amides derived from
ammonia, primary C.sub.1-C.sub.6 alkyl amines and secondary
C.sub.1-C.sub.6 dialkyl amines wherein the alkyl groups are
straight or branched chain. In the case of secondary amines the
amine may also be in the form of a 5- or 6-membered heterocycle
containing one nitrogen atom. Amides derived from ammonia,
C.sub.1-C.sub.3 alkyl primary amines, and C.sub.1-C.sub.2 dialkyl
secondary amines are preferred. Amides of the compounds of the
invention may be prepared according to conventional methods.
[0093] 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, "Pro-drugs as
Novel Delivery Systems," Vol. 14 of the A.C.S. Symposium Series,
and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche,
American Pharmaceutical Association and Pergamon Press, 1987
(incorporated herein by reference. In general, a prodrug is a drug
which has been chemically modified and may be biologically inactive
at its site of action, but which may be degraded or modified by one
or more enzymatic or other in vivo processes to the parent
bioactive form.
[0094] A "therapeutically effective amount" is an amount of a
compound of Formula I that when administered to a patient,
ameliorates a symptom of the disease.
[0095] The BCAT inhibitor, which is preferably a compound of
Formula I, a BCAT pathway-inhibiting analogue of Formula I, a BCAT
pathway-inhibiting prodrug of Formula I, or a pharmaceutically
acceptable salt of any of the foregoing, can be administered in
accordance with the present inventive method by any suitable route.
Suitable routes of administration include systemic, such as orally
or by injection, topical, intraocular, periocular (e.g.,
subTenon's), subconjunctival, subretinal, suprachoroidal and
retrobulbar. The manner in which the BCAT inhibitor is administered
is dependent, in part, upon whether the treatment of retinopathy is
prophylactic or therapeutic. The manner in which the BCAT inhibitor
is administered for therapeutic treatment of retinopathy is
dependent, in part, upon the cause of the retinopathy.
[0096] For example, given that diabetes is the leading cause of
retinopathy, the BCAT inhibitor can be administered
prophylactically as soon as the pre-diabetic retinopathy state is
detected. For the prophylactic treatment of retinopathy that can
result from diabetes, the BCAT inhibitor is preferably administered
systemically, e.g., orally or by injection. For the therapeutic
treatment of nonproliferative diabetic retinopathy, the BCAT
inhibitor can be administered systemically, e.g., orally or by
injection, or intraocularly. Proliferative diabetic retinopathy can
be therapeutically treated by the administration of the BCAT
inhibitor intraocularly, topically, subconjunctivally or
periocularly (e.g., subTenon's), for example. The BCAT inhibitor is
preferably administered intraocularly, topically, subconjunctivally
or periocularly (e.g., subTenon's) for the prophylactic or
therapeutic treatment of retinopathy before, during or after
surgical removal from an eye of scar tissue generated during
neovascularization during the proliferative diabetic stage.
[0097] The BCAT inhibitor is preferably administered as soon as
possible after it has been determined that an animal, such as a
mammal, specifically a human, is at risk for retinopathy
(prophylactic treatment) or has begun to develop retinopathy
(therapeutic treatment). Treatment will depend, in part, upon the
particular BCAT inhibitor used, the amount of the BCAT inhibitor
administered, the route of administration, and the cause and
extent, if any, of retinopathy realized.
[0098] One skilled in the art will appreciate that suitable methods
of administering a BCAT inhibitor, which is useful in the present
inventive methods, are available. Although more than one route can
be used to administer a particular BCAT inhibitor, a particular
route can provide a more immediate and more effective reaction than
another route. Accordingly, the described routes of administration
are merely exemplary and are in no way limiting.
[0099] The dose administered to an animal, particularly a human, in
accordance with the present invention should be sufficient to
effect the desired response in the animal over a reasonable time
frame. One skilled in the art will recognize that dosage will
depend upon a variety of factors, including the strength of the
particular BCAT inhibitor employed, the age, species, condition or
disease state, and body weight of the animal, as well as the amount
of the retina about to be affected or actually affected by
retinopathy. The size of the dose also will be determined by the
route, timing and frequency of administration as well as the
existence, nature, and extent of any adverse side effects that
might accompany the administration of a particular BCAT inhibitor
and the desired physiological effect. It will be appreciated by one
of ordinary skill in the art that various conditions or disease
states, in particular, chronic conditions or disease states, may
require prolonged treatment involving multiple administrations.
[0100] Suitable doses and dosage regimens can be determined by
conventional range-finding techniques known to those of ordinary
skill in the art. Generally, treatment is initiated with smaller
dosages, which are less than the optimum dose of the compound.
Thereafter, the dosage is increased by small increments until the
optimum effect under the circumstances is reached. The present
inventive method will typically involve the administration of from
about 1 mg/kg/day to about 100 mg/kg/day, preferably from about 15
mg/kg/day to about 50 mg/kg/day, if administered systemically.
Intraocular administration typically will involve the
administration of from about 0.1 mg total to about 5 mg total,
preferably from about 0.5 mg total to about 1 mg total. A preferred
concentration for topical administration is 100 mu M.
[0101] Compositions for use in the present inventive method
preferably comprise a pharmaceutically acceptable carrier and an
amount of a BCAT inhibitor sufficient to treat retinopathy
prophylactically or therapeutically. The carrier can be any of
those conventionally used and is limited only by chemico-physical
considerations, such as solubility and lack of reactivity with the
compound, and by the route of administration. It will be
appreciated by one of ordinary skill in the art that, in addition
to the following described pharmaceutical compositions, the BCAT
inhibitor can be formulated as polymeric compositions, inclusion
complexes, such as cyclodextrin inclusion complexes, liposomes,
microspheres, microcapsules and the like (see, e.g., U.S. Pat. Nos.
4,997,652; 5,185,152; and 5,718,922).
[0102] The BCAT inhibitor can be formulated as a pharmaceutically
acceptable acid addition salt. Examples of pharmaceutically
acceptable acid addition salts for use in the pharmaceutical
composition include those derived from mineral acids, such as
hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric and
sulfuric acids, and organic acids, such as tartaric, acetic,
citric, malic, lactic, fumaric, benzoic, glycolic, gluconic,
succinic, and arylsulphonic, for example p-toluenesulphonic,
acids.
[0103] The pharmaceutically acceptable excipients described herein,
for example, vehicles, adjuvants, carriers or diluents, are
well-known to those who are skilled in the art and are readily
available to the public. It is preferred that the pharmaceutically
acceptable carrier be one which is chemically inert to the BCAT
inhibitor and one which has no detrimental side effects or toxicity
under the conditions of use.
[0104] The choice of excipient will be determined in part by the
particular BCAT inhibitor, as well as by the particular method used
to administer the composition. Accordingly, there is a wide variety
of suitable formulations of the pharmaceutical composition of the
present invention. The following formulations are merely exemplary
and are in no way limiting.
[0105] Injectable formulations are among those that are preferred
in accordance with the present inventive method. The requirements
for effective pharmaceutically carriers for injectable compositions
are well-known to those of ordinary skill in the art, (see
Pharmaceutics and Pharmacy Practice, J. B. Lippincott Co.,
Philadelphia, Pa., Banker and Chalmers, eds., pages 238-250 (1982),
and ASHP Handbook on Injectable Drugs, Toissel, 4th ed., pages
622-630 (1986). It is preferred that such injectable compositions
be administered intramuscularly, intravenously, or
intraperitoneally.
[0106] Topical formulations are well-known to those of skill in the
art. Such formulations are suitable in the context of the present
invention for application to the skin. The use of patches, corneal
shields (see, e.g., U.S. Pat. No. 5,185,152), and ophthalmic
solutions (see, e.g., U.S. Pat. No. 5,710,182) and ointments, e.g.,
eye drops, is also within the skill in the art.
[0107] Formulations suitable for oral administration can consist of
(a) liquid solutions, such as an effective amount of the compound
dissolved in diluents, such as water, saline, or orange juice; (b)
capsules, sachets, tablets, lozenges, and troches, each containing
a predetermined amount of the active ingredient, as solids or
granules; (c) powders; (d) suspensions in an appropriate liquid;
and (e) suitable emulsions. Liquid formulations may include
diluents, such as water and alcohols, for example, ethanol, benzyl
alcohol, and the polyethylene alcohols, either with or without the
addition of a pharmaceutically acceptable surfactant, suspending
agent, or emulsifying agent. Capsule forms can be of the ordinary
hard- or soft-shelled gelatin type containing, for example,
surfactants, lubricants, and inert fillers, such as lactose,
sucrose, calcium phosphate, and corn starch. Tablet forms can
include one or more of lactose, sucrose, mannitol, corn starch,
potato starch, alginic acid, microcrystalline cellulose, acacia,
gelatin, guar gum, colloidal silicon dioxide, croscarmellose
sodium, talc, magnesium stearate, calcium stearate, zinc stearate,
stearic acid, and other excipients, colorants, diluents, buffering
agents, disintegrating agents, moistening agents, preservatives,
flavoring agents, and pharmacologically compatible excipients.
Lozenge forms can comprise the active ingredient in a flavor,
usually sucrose and acacia or tragacanth, as well as pastilles
comprising the active ingredient in an inert base, such as gelatin
and glycerin, or sucrose and acacia, emulsions, gels, and the like
containing, in addition to the active ingredient, such excipients
as are known in the art.
[0108] The effectiveness of an orally administered drug is
dependent upon the drug's efficient transport across the mucosal
epithelium and its stability in entero-hepatic circulation. Drugs
that are effective after parenteral administration but less
effective orally, or whose plasma half-life is considered too
short, may be chemically modified into a prodrug form.
[0109] Formulations suitable for parenteral administration include
aqueous and non-aqueous, isotonic sterile injection solutions,
which can contain anti-oxidants, buffers, bacteriostats, and
solutes that render the formulation isotonic with the blood of the
intended recipient, and aqueous and non-aqueous sterile suspensions
that can include suspending agents, solubilizers, thickening
agents, stabilizers, and preservatives. The inhibitor can be
administered in a physiologically acceptable diluent in a
pharmaceutical carrier, such as a sterile liquid or mixture of
liquids, including water, saline, aqueous dextrose and related
sugar solutions, an alcohol, such as ethanol, isopropanol, or
hexadecyl alcohol, glycols, such as propylene glycol or
polyethylene glycol, dimethylsulfoxide, glycerol ketals, such as
2,2-dimethyl-1,3-dioxolane-4-- methanol, ethers, such as
poly(ethyleneglycol) 400, an oil, a fatty acid, a fatty acid ester
or glyceride, or an acetylated fatty acid glyceride, with or
without the addition of a pharmaceutically acceptable surfactant,
such as a soap or a detergent, suspending agent, such as pectin,
carbomers, methylcellulose, hydroxypropylmethylcellulose, or
carboxymethylcellulose, or emulsifying agents and other
pharmaceutical adjuvants. Oils, which can be used in parenteral
formulations include petroleum, animal, vegetable, or synthetic
oils. Specific examples of oils include peanut, soybean, sesame,
cottonseed, corn, olive, petrolatum, and mineral.
[0110] Suitable fatty acids for use in parenteral formulations
include oleic acid, stearic acid, and isostearic acid. Ethyl oleate
and isopropyl myristate are examples of suitable fatty acid
esters.
[0111] Suitable soaps for use in parenteral formulations include
fatty alkali metals, ammonium, and triethanolamine salts, and
suitable detergents include (a) cationic detergents such as, for
example, dimethyl dialkyl ammonium halides, and alkyl pyridinium
halides, (b) anionic detergents such as, for example, alkyl, aryl,
and olefin sulfonates, alkyl, olefin, ether, and monoglyceride
sulfates, and sulfosuccinates, (c) nonionic detergents such as, for
example, fatty amine oxides, fatty acid alkanolamides, and
polyoxyethylenepolypropylene copolymers, (d) amphoteric detergents
such as, for example, alkyl-p-aminopropionates, and
2-alkyl-imidazoline quaternary ammonium salts, and (e) mixtures
thereof.
[0112] The parenteral formulations will typically contain from
about 0.5 to about 25% by weight of the active ingredient in
solution. Preservatives and buffers may be used. In order to
minimize or eliminate irritation at the site of injection, such
compositions may contain one or more nonionic surfactants having a
hydrophile-lipophile balance (HLB) of from about 12 to about
17.
[0113] The quantity of surfactant in such formulations will
typically range from about 5 to about 15% by weight. Suitable
surfactants include polyethylene sorbitan fatty acid esters, such
as sorbitan monooleate and the high molecular weight adducts of
ethylene oxide with a hydrophobic base, formed by the condensation
of propylene oxide with propylene glycol. The parenteral
formulations can be presented in unit-dose or multi-dose sealed
containers, such as ampules and vials, and can be stored in a
freeze-dried (lyophilized) condition requiring only the addition of
the sterile liquid excipient, for example, water, for injections,
immediately prior to use. Extemporaneous injection solutions and
suspensions can be prepared from sterile powders, granules, and
tablets of the kind previously described.
[0114] Such compositions can be formulated as intraocular
formulations, sustained-release formulations or devices (see, e.g.,
U.S. Pat. No. 5,378,475). For example, gelantin, chondroitin
sulfate, a polyphosphoester, such as
bis-2-hydroxyethyl-terephthalate (BHET), or a polylactic-glycolic
acid (in various proportions) can be used to formulate
sustained-release formulations. Implants (see, e.g., U.S. Pat. Nos.
5,443,505; 4,853,224; and 4,997,652), devices (see, e.g., U.S. Pat.
Nos. 5,554,187; 4,863,457; 5,098,443; and 5,725,493), such as an
implantable device, e.g., a mechanical reservoir, an intraocular
device or an extraocular device with an intraocular conduit (e.g.,
100 mu-1 mm in diameter), or an implant or a device comprised of a
polymeric composition as described above, can be used.
[0115] The present inventive method also can involve the
co-administration of other pharmaceutically active compounds. By
"co-administration" is meant administration before, concurrently
with, e.g., in combination with the BCAT inhibitor in the same
formulation or in separate formulations, or after administration of
a BCAT inhibitor as described above. For example, corticosteroids,
e.g., prednisone, methylprednisolone, dexamethasone, or
triamcinalone acetinide, or noncorticosteroid anti-inflammatory
compounds, such as ibuprofen or flubiproben, can be
co-administered. Similarly, vitamins and minerals, e.g., zinc,
antioxidants, e.g., carotenoids (such as a xanthophyll carotenoid
like zeaxanthin or lutein), and micronutrients can be
co-administered. A general synthetic scheme for preparing compounds
of Formula I is set forth below. 2
EXAMPLE 1
[0116] Benzoic acid, 4-nitro-2-(phenylsulfonyl)hydrazide 3
[0117] A solution of 4-nitro benzoic acid (0.2 g, 1.2 mmol in 10 mL
of dry THF) was cooled to 0.degree. C., treated with
N-methyl-morpholine (1.6 mL, 14.6 mmol) as well as
isobutyl-chloroformate (1.7 mL, 1.3 mmol), and stirred for 10
minutes prior to the addition of benzenesulfonyl hydrazide (0.21 g,
1.2 mmol). After the addition of hydrazide, the mixture was stirred
for 3 hours at 25.degree. C. Then the reaction was diluted with
ethyl acetate (100 mL), washed with saturated sodium bicarbonate
solution as well as brine, and concentrated in vacuum. The crude
material was chromatographed on silica gel eluting with
hexanes/ethyl acetate=2:1 to give the desired product (65%
yield).
[0118] MS: 322.0 (M+1 for C.sub.13H.sub.11N.sub.3O.sub.5S.sub.1);
mp 193-195.degree. C.; TLC (SiO.sub.2) R.sub.f=0.74 (8%
MeOH/CH.sub.2Cl.sub.2). HPLC (C18 column, 1:1
CH.sub.3CN/H.sub.2O+0.1% TFA) 94.95%, RT=5.045 min. HRMS (calc for
M+1) 321.0419 (found) 321.0421. IR (KBr, cm.sup.-1): 3287, 3067,
2822, 1673, 1520, 1340, 1160, 978. .sup.1H NMR (DMSO-d.sub.6)
.delta. 7.48-7.52 (m, 2H), 7.58-7.62 (m, 1H), 7.80-7.87 (m, 4H),
8.23-8.26 (m, 2H), 10.18 (s, 1H), 11.01 (s, 1H).
EXAMPLE 2
[0119] Benzoic acid, 4-amino-2-(phenylsulfonyl)hydrazide 4
[0120] Synthesis of Example 2: Benzoic acid,
4-nitro-2-(phenylsulfonyl)hyd- razide (0.82 g, Example 1) was
dissolved in 50 mL of THF/EtOH (1:1), treated with wet Raney Nickel
(0.6 g) and hydrogenated at 50 psi for 15 hours. The reaction was
filtered, concentrated in vacuum, and chromatographed on silica gel
eluting with 6% MeOH/CH.sub.2Cl.sub.2 to give 61 mg (84%) of the
desired product.
[0121] MS: 291.9 (M+1 for C.sub.13H.sub.13N.sub.3O.sub.3S.sub.1);
mp 171-173.degree. C.; TLC (SiO.sub.2) R.sub.f=0.45 (8%
MeOH/CH.sub.2Cl.sub.2); HPLC (C18 column, 1:1
CH.sub.3CN/H.sub.2O+0.1% TFA) 99.14%, RT=2.575 min. HRMS (calc for
M+1) 292.0756 (found) 292.0753. IR (KBr, cm.sup.-1): 3471, 3367,
3151, 2925, 1661, 1318, 1162. .sup.1H NMR (DMSO-d.sub.6) .delta.
5.69 (s, 2H), 6.44 (d, 2H, J=8.5 Hz), 7.32-7.58 (m, 5H), 7.75-7.80
(m, 2H), 9.68 (br, 1H), 10.12 (br, 1H).
EXAMPLE 3
[0122] Benzoic acid, 4-[[(2,4,6-trichlorophenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 5
[0123] Benzoic acid, 4-amino-2-(phenylsulfonyl)hydrazide (0.15 g,
0.52 mmol, Example 2) was dissolved in 7 mL of dry pyridine, then
2,4,6-(trichlorophenyl) sulfonyl chloride (0.15 mL, 0.52 mmol) was
added, and the reaction was stirred at 25.degree. C. for overnight.
The solvent was removed, and the product was recrystalized from
ethyl acetate and hexane to give 0.26 g (96%) of the product as a
white solid.
[0124] MS: 536.0 (M+1 for
C.sub.19H.sub.14Cl.sub.3N.sub.3O.sub.5S.sub.2); mp
195.7-196.2.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA) 96.3%, RT=8.0 min. .sup.1H NMR (DMSO-D.sub.6) .delta. 7.10 (d,
1H, J=6.8 Hz), 7.40-7.60 (m, 7H), 7.75 (d, 1H, J=7.5 Hz), 7.79 (S,
1H), 8.63 (S, 1H), 9.92 (S, 1H), 10.49 (S, 1H), 11.35 (S, 1H).
EXAMPLE 4
[0125] Benzoic acid,
4-[[[4-(trifluoromethyl)phenyl]sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 6
[0126] Example 4 was synthesized in accordance with the methods of
Example 3 except that 4-[(trifluoromethyl)phenyl]sulfonyl chloride
was used instead of 2,4,6-(trichlorophenyl)sulfonyl chloride (yield
92%).
[0127] MS: 500.0 (M+1 for
C.sub.20H.sub.16F.sub.3N.sub.3O.sub.5S.sub.2); mp 98.5-99.4.degree.
C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1% TFA) 96.8%, RT=6.1
min. .sup.1H NMR(DMSO-D.sub.6) .delta. 7.10 (d, 1H, J=8.6 Hz),
7.40-7.60 (m, 5H), 7.76 (d, 1H, J=7.5 Hz), 7.86-8.06 (m, 5H), 8.63
(d, 1H, b J=4 Hz), 9.94 (S, 1H), 10.52 (S, 1H), 10.95 (S, 1H).
EXAMPLE 5
[0128] Benzoic acid, 4-[[(3,4-dimethoxyphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 7
[0129] Example 5 was synthesized in accordance with the methods of
Example 3 except that (3,4-dimethoxypheny)sulfonyl chloride was
used instead of 2,4,6-(trichlorophenyl)sulfonyl chloride (yield
92%).
[0130] MS: 492.0 (M+1 for C.sub.21H.sub.21N.sub.3O.sub.7S.sub.2);
mp 221.9-222.6.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA) 92.02%, RT=3.5 min. .sup.1H NMR (DMSO-D.sub.6) .delta. 3.72
(S, 3H), 3.75 (S, 3H), 7.01 (d, 1H, J=8.6 Hz), 7.08 (d, 2H, J=8.5
Hz), 7.25 (S, 1H), 7.32 (d, 1H, J=6.9 Hz), 7.40-7.60 (m, 5H), 7.75
(d, 2H, J=7.6 Hz), 9.91 (S, 1H), 10.48 (S, 1H), 10.52 (S, 1H).
EXAMPLE 6
[0131] Benzoic acid, 4-[[[(4-phenoxy)benzene]sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 8
[0132] Example 6 was synthesized in accordance with the methods of
Example 3 except that [(4-phenoxy)benzene]sulfonyl chloride was
used instead of 2,4,6-(trichlorophenyl)sulfonyl chloride (yield
96%).
[0133] MS: 524.1 (M+1 for C.sub.25H.sub.21N.sub.3O.sub.6S.sub.2);
mp 94.1-94.9.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA, 85.08%, RT=8.7 min. .sup.1H NMR (DMSO-D.sub.6) .delta. 7.01
(d, 2H, J=4.8 Hz), 7.09 (m, 2H), 7.40-7.60 (m, 10H), 7.78 (m, 4H),
9.93 (S, 1H), 10.51 (S, 1H), 10.68 (S, 1H).
EXAMPLE 7
[0134] Benzoic acid, 4-[[(2-chloro-6-methylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 9
[0135] Example 7 was synthesized in accordance with the methods of
Example 3 except that (2-chloro-6-methylphenyl)sulfonyl chloride
was used instead of 2,4,6-(trichlorophenyl)sulfonyl chloride (yield
91%).
[0136] MS: 480.1 (M+1 for C.sub.20H.sub.18ClN.sub.3O.sub.5S.sub.2);
mp 214.4-215.6.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA) 92.5%, RT=6.3 min. CHN (calc) C, 50.05; H, 3.78; N, 8.7
(found) C, 50.18; H, 3.83; N, 8.59). .sup.1H NMR (DMSO-d.sub.6)
.delta. 2.65 (S, 3H), 7.02 (d, 2H, J=8.8 Hz), 7.34 (t, 1H, J=4.9
Hz), 7.40-7.60 (m, 7H), 7.75 (d, 2H, J=7.3 Hz), 9.89 (S, 1H), 10.44
(S, 1H), 10.93 (S, 1H).
EXAMPLE 8
[0137] Benzoic acid, 4-[(methylsulfonyl)amino]-,
2-(phenylsulfonyl)hydrazi- de 10
[0138] Example 8 was synthesized in accordance with the methods of
Example 3 except that methylsulfonyl chloride was used instead of
2,4,6-(trichlorophenyl)-sulfonyl chloride (yield 94%).
[0139] MS: 370.0 (M+1 for C.sub.14H.sub.15ClN.sub.3O.sub.5S.sub.2);
mp 185.6-186.5.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA) 89.4%, RT=2.8 min. .sup.1H NMR (DMSO-d.sub.6) .delta. 2.91 (S,
3H), 7.18 (d, 2H, J=7.1 Hz), 7.40 (t, 2H, J=6.8 Hz), 7.50-7.60 (m,
1H), 7.63 (d, 2H, J=7.9 Hz), 7.85 (d, 2H, J=8.1 Hz), 8.88 (S, 1H),
9.89 (S, 1H), 10.63 (S, 1H).
EXAMPLE 9
[0140] Benzoic acid, 4-[[(4-methylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 11
[0141] Example 9 was synthesized in accordance with the methods of
Example 3 except that (4-methylphenyl)sulfonyl chloride was used
instead of 2,4,6-(trichlorophenyl)sulfonyl chloride (yield
96%).
[0142] MS: 446.1 (M+1 for C.sub.20H.sub.19N.sub.3O.sub.5S.sub.2);
mp 191.6-192.4.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA) 95.2%, RT=4.7 min. .sup.1H NMR (DMSO-d.sub.6) .delta. 2.91 (S,
3H), 7.10 (d, 2H, J=6.8 Hz), 7.32 (d, 2H, J=5.8 Hz), 7.40-7.60 (m,
5H), 7.66 (d, 2H, J=6.1 Hz), 7.75 (d, 2H, J=8.1 Hz), 9.91 (S, 1H),
10.48 (S, 1H), 10.64 (S, 1H).
EXAMPLE 10
[0143] Benzoic acid, 4-[[(4-ethylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 12
[0144] Example 10 was synthesized in accordance with the methods of
Example 3 except that (4-ethylbenzene)sulfonyl chloride was used
instead of 2,4,6-(trichlorophenyl)sulfonyl chloride (yield
92%).
[0145] MS: 460.1 (M+1 for C.sub.21H.sub.21N.sub.3O.sub.5S.sub.2);
mp 142.3-142.9.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA) 93.5%, RT=5.9 min. .sup.1H NMR (DMSO-d.sub.6) .delta. 1.11 (t,
3H, J=7.6 Hz), 2.59 (q, 2H, J=7.5 Hz), 7.10 (d, 2H, J=8.6 Hz), 7.35
(d, 2H, J=8.0 Hz), 7.40-7.60 (m, 5H), 7.68 (d, 2H, J=8.1 Hz), 7.75
(d, 2H, J=7.5 Hz), 9.91 (S, 1H), 10.48 (S, 1H), 10.68 (S, 1H).
EXAMPLE 11
[0146] Benzoic acid, 4-[[(4-n-propylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 13
[0147] Example 11 was synthesized in accordance with the methods of
Example 3 except that (4-n-propylbenzene)sulfonyl chloride was used
instead of 2,4,6-(trichlorophenyl)sulfonyl chloride (yield
90%).
[0148] MS: 474.1 (M+1 for C.sub.22H.sub.23N.sub.3O.sub.5S.sub.2);
HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1% TFA) 95.3%, RT=7.4 min.
.sup.1H NMR (DMSO-d.sub.6) .delta. 0.79 (t, 3H, J=7.5 Hz), 1.25 (M,
2H), 2.59 (q, 2H, J=7.5 Hz), 7.10 (d, 2H, J=8.6 Hz), 7.35 (d, 2H,
J=8.0 Hz), 7.40-7.60 (m, 5H), 7.68 (d, 2H, J=8.1 Hz), 7.75 (d, 2H,
J=7.5 Hz), 9.91 (S, 1H), 10.46 (S, 1H), 10.68 (S, 1H).
EXAMPLE 12
[0149] Benzoic acid, 4-[[(4-n-butylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 14
[0150] Example 12 was synthesized in accordance with the methods of
Example 3 except that (4-n-butylbenzene)sulfonyl chloride was used
instead of 2,4,6-(trichlorophenyl)-sulfonyl chloride (yield
92%).
[0151] MS: 488.1 (M+1 for C.sub.23H.sub.25N.sub.3O.sub.5S.sub.2);
mp 76.0-77.1.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA) 94.0%, RT=9.9 min. .sup.1H NMR (DMSO-d.sub.6) .delta. 0.82 (t,
3H, J=7.4 Hz), 1.22 (m, 2H, J=7.6 Hz), 1.48 (m, 2H, J=7.8 Hz), 2.56
(t, 2H, J=7.6 Hz), 7.10 (d, 2H, J=8.8 Hz), 7.35 (d, 2H, J=8.1 Hz),
7.40-7.60 (m, 5H), 7.67 (d, 2H, J=8.3 Hz), 7.76 (d, 2H, J=7.6 Hz),
9.91 (S, 1H), 10.49 (S, 1H), 10.66 (S, 1H).
EXAMPLE 13
[0152] Benzoic acid, 4-[[(4-n-pentylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 15
[0153] Example 13 was synthesized in accordance with the methods of
Example 3 except that (4-n-pentylbenzene)sulfonyl chloride was used
instead of 2,4,6-(trichlorophenyl)-sulfonyl chloride (yield
90%).
[0154] MS: 502.1 (M+1 for C.sub.24H.sub.27N.sub.3O.sub.5S.sub.2);
mp 97.1-97.9.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA) 95.0%, RT=1.5 min. .sup.1H NMR (DMSO-D.sub.6) .delta. 0.85 (t,
3H, J=6.6 Hz), 1.28 (m, 4H, J=6.6 Hz), 1.56 (m, 2H, J=7.1 Hz), 2.59
(t, 2H, J=7.6 Hz), 7.07 (d, 2H, J=8.6 Hz), 7.24 (d, 2H, J=9.5 Hz),
7.41 (t, 2H, J=7.8 Hz), 7.50-7.70 (m, 3H), 7.71 (d, 2H, J=8.3 Hz),
7.92 (d, 2H, J=8.3 Hz), 9.90 (S, 1H), 10.46 (S, 1H), 10.64 (S,
1H).
EXAMPLE 14
[0155] Benzoic acid, 4-[[(4-fluorophenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 16
[0156] Example 14 was synthesized in accordance with the methods of
Example 3 except that (4-fluorobenzene)sulfonyl chloride was used
instead of 2,4,6-(trichlorophenyl)-sulfonyl chloride.
[0157] MS: 450.1 (M+1 for C.sub.19H.sub.16FN.sub.3O.sub.5S.sub.2);
mp 183.5-185.0.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA) 97.9%, RT=4.4 min. CHN (calc) C, 50.77; H, 3.59; N, 9.35
(found) C, 50.58; H, 3.65; N, 9.05. .sup.1H NMR (DMSO-d.sub.6)
.delta. 7.09 (d, 2H, J=8.6 Hz), 7.37 (t, 2H, J=6.6 Hz), 7.46 (t,
2H, J=7.9 Hz), 7.58 (t, 2H, J=4.1 Hz), 7.76 (d, 2H, J=3.1 Hz),
7.80-7.90 (m, 3H), 9.92 (S, 1H), 10.49 (S, 1H), 10.72 (S, 1H).
EXAMPLE 15
[0158] Benzoic acid, 4-[(phenylsulfonyl)]amino]-,
2-(phenylsulfonyl)hydraz- ide 17
[0159] Example 15 was synthesized in accordance with the methods of
Example 3 except that benzenesulfonyl chloride was used instead of
2,4,6-(trichlorophenyl)sulfonyl chloride.
[0160] MS: 432.0 (M+1 for C.sub.19H.sub.17N.sub.3O.sub.5S.sub.2);
mp 140.8-144.5.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA) 97.9%, RT=4.2 min. CHN (calc) C, 52.89; H, 3.97; N, 9.74
(found) C, 53.41; H, 3.95; N, 9.77. .sup.1H NMR (DMSO-d.sub.6)
.delta. 7.20 (d, 2H, J=8.8 Hz), 7.50-7.70 (m, 8H), 7.80-7.95 (m,
4H), 10.01 (S, 1H), 10.58 (S, 1H), 10.82 (S, 1H).
EXAMPLE 16
[0161] 4-Benzylamino-benzoic acid 2-(phenylsulfonyl)hydrazide
18
[0162] 4-Aminobenzoic acid 2-(phenylsulfonyl)hydrazide (100 mg,
0.34 mmol) was dissolved in THF (3.4 mL), treated with acetic acid
(20 .mu.L, 0.34 mmol) and benzaldehyde (23 .mu.L, 0.34 mmol), and
stirred for 5 minutes. Sodium triacetoxyborohydride (87 mg, 0.41
mmol) was added, and the reaction was stirred overnight. The
reaction was diluted with EtOAc (100 mL), washed with saturated
sodium bicarbonate solution and brine, dried over Na.sub.2SO.sub.4,
and concentrated in vacuo. The residue was chromatographed on
silica gel eluting with 3:2 hexanes/EtOAc to give 23 mg (18%) of
the desired product.
[0163] MS: 382.0 (M+1 for C.sub.20H.sub.19N.sub.3O.sub.3S.sub.1);
mp 214-215.degree. C.; TLC (SiO.sub.2) R.sub.f=0.43 (1:1
hexane/EtOAc); HPLC (C18 column, 1:1 CH.sub.3CN/H.sub.2O+0.1% TFA)
99.04%, RT=8.309 min. HRMS (calc) 382.1225 (found) 382.1222; IR
(KBr, cm.sup.-1): 3404, 3355, 3107, 1657, 1607, 1159. .sup.1H NMR
(DMSO-d.sub.6) .delta. 4.25 (d, 2H, J=5.6 Hz), 6.47 (d, 2H, J=8.3
Hz), 6.86-6.88 (m, 1H), 7.26-7.28 (m, 4H), 7.37-7.40 (m, 2H),
7.42-7.45 (m, 2H), 7.52-7.56 (m, 1H), 7.74 (d, 2H, J=7.8 Hz), 9.58
(s, 1H) 9.68 (s, 1H), 10.13 (s, 1H).
EXAMPLE 17
[0164] Benzoic acid, 4-[[(3-methylphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 19
[0165] Example, 17 was synthesized in accordance with the methods
of Example 3 except that (3-methylphenyl)sulfonyl chloride was used
instead of 2,4,6-(trichlorophenyl)sulfonyl chloride (yield
96%).
[0166] MS: 446.1 (M+1 for C.sub.20H.sub.19N.sub.3O.sub.5S.sub.2);
mp 219.0-219.7.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O+0.1%
TFA) 98.8%, RT=5.2 min. .sup.1H NMR (DMSO-d.sub.6) .delta. 2.31 (S,
3H), 7.09 (d, 2H, J=8.6 Hz), 7.40-7.60 (m, 9H), 7.75 (d, 2H, J=7.3
Hz), 9.91 (S, 1H), 10.48 (S, 1H), 10.68 (S, 1H).
EXAMPLE 18
[0167] 4-Phenoxybenzoic acid, 2-[(3-methyl)phenylsulfonyl]hydrazide
20
[0168] A solution of 2-phenoxybenzoic acid hydrazide (0.3 M) in
pyridine (0.513 mL) was treated with a solution of the
3-(methylphenyl)sulfonyl chloride (1.0 M) in CH.sub.2Cl.sub.2
(0.225 mL). The reaction was placed in a Bohdan Miniblock apparatus
and shaken at 45.degree. C. for 16 hours. The reaction was cooled
to room temperature, treated with polymer-supported polyamine
quench resin (Aldrich, 100 mg), and shaken for 16 hours. The
solution was filtered and concentrated. The residue was purified by
preparative HPLC on a Phenomenex Develofil 28.times.100 mm C-18
column eluting with a gradient of 10% to 100%
CH.sub.3CN/H.sub.2O+3% n-propanol.
[0169] MS: 384 (M+1 for C.sub.20H.sub.18N.sub.2O.sub.4S); HPLC
(Waters Alliance 2790 column, solvent gradient of 60% to 100%
CH.sub.3CN/H.sub.2O+0.1% formic acid. Purity: 100%.
EXAMPLE 19
[0170] 4-Phenoxybenzoic acid, 2-[3-bromophenylsulfonyl]hydrazide
21
[0171] A solution of 2-phenoxybenzoic acid hydrazide (0.3 M) in
pyridine (0.513 mL) was treated with a solution of the
3-(bromophenyl)sulfonyl chloride (1.0 M) in CH.sub.2Cl.sub.2 (0.225
mL). The reaction was placed in a Bohdan Miniblock apparatus and
shaken at 45.degree. C. for 16 hours. The reaction was cooled to
room temperature, treated with polymer-supported polyamine quench
resin (Aldrich, 100 mg), and shaken for 16 hours. The solution was
filtered and concentrated. The residue was purified by preparative
HPLC on a Phenomenex Develofil 28.times.100 mm C-18 column eluting
with a gradient of 10% to 100% CH.sub.3CN/H.sub.2O+3%
n-propanol.
[0172] MS: 448 (M+1 for C.sub.19H.sub.15BrN.sub.2O.sub.4S); HPLC
(Waters Alliance 2790 column, solvent gradient of 60% to 100%
CH.sub.3CN/H.sub.2O+0.1% formic acid. Purity: 96%.
EXAMPLE 20
[0173] 4-Phenoxybenzoic acid, 2-[2-chlorophenylsulfonyl]hydrazide
22
[0174] A solution of 2-phenoxybenzoic acid hydrazide (0.3 M) in
pyridine (0.513 mL) was treated with a solution of the
2-(chlorophenyl)sulfonyl chloride (1.0 M) in CH.sub.2Cl.sub.2
(0.225 mL). The reaction was placed in a Bohdan Miniblock apparatus
and shaken at 45.degree. C. for 16 hours. The reaction was cooled
to room temperature, treated with polymer-supported polyamine
quench resin (Aldrich, 100 mg), and shaken for 16 hours. The
solution was filtered and concentrated. The residue was purified by
preparative HPLC on a Phenomenex Develofil 28.times.100 mm C-18
column eluting with a gradient of 10% to 100%
CH.sub.3CN/H.sub.2O+3% n-propanol.
[0175] MS: 403 (M+1 for C.sub.19H.sub.15ClN.sub.2O.sub.4S); HPLC
(Waters Alliance 2790 column, solvent gradient of 60% to 100%
CH.sub.3CN/H.sub.2O+0.1% formic acid. Purity: 98%.
EXAMPLE 21
[0176] 4-Phenoxybenzoic acid,
2-[2-(trifluoromethoxy)-phenylsulfonyl]hydra- zide 23
[0177] A solution of 2-phenoxybenzoic acid hydrazide (0.3 M) in
pyridine (0.513 mL) was treated with a solution of the
2-(trifluoromethoxy)phenyls- ulfonyl chloride (1.0 M) in
CH.sub.2Cl.sub.2 (0.225 mL). The reaction was placed in a Bohdan
Miniblock apparatus and shaken at 45.degree. C. for 16 hours. The
reaction was cooled to room temperature, treated with
polymer-supported polyamine quench resin (Aldrich, 100 mg), and
shaken for 16 hours. The solution was filtered and concentrated.
The residue was purified by preparative HPLC on a Phenomenex
Develofil 28.times.100 mm C-18 column eluting with a gradient of
10% to 100% CH.sub.3CN/H.sub.2O+3% n-propanol.
[0178] MS: 453 (M+1 for C.sub.20H.sub.15F.sub.3N.sub.2O.sub.5S);
HPLC (Waters Alliance 2790 column, solvent gradient of 60% to 100%
CH.sub.3CN/H.sub.2O+0.1% formic acid. Purity: 95%.
EXAMPLE 22
[0179] 4-Phenoxybenzoic acid,
2-[(4-methyl-3-nitro)phenylsulfonyl]hydrazid- e 24
[0180] A solution of 2-phenoxybenzoic acid hydrazide (0.3 M) in
pyridine (0.513 mL) was treated with a solution of the
(4-methyl-3-nitro)phenylsul- fonyl chloride (1.0 M) in
CH.sub.2Cl.sub.2 (0.225 mL). The reaction was placed in a Bohdan
Miniblock apparatus and shaken at 45.degree. C. for 16 hours. The
reaction was cooled to room temperature, treated with
polymer-supported polyamine quench resin (Aldrich, 100 mg), and
shaken for 16 hours. The solution was filtered and concentrated.
The residue was purified by preparative HPLC on a Phenomenex
Develofil 28.times.100 mm C-18 column eluting with a gradient of
10% to 100% CH.sub.3CN/H.sub.2O+3% n-propanol.
[0181] MS: 428 (M+1 for C.sub.10H.sub.17N.sub.3O.sub.6S); HPLC
(Waters Alliance 2790 column, solvent gradient of 60% to 100%
CH.sub.3CN/H.sub.2O+0.1% formic acid. Purity: 94%.
EXAMPLE 23
[0182] 4-Phenoxybenzoic acid,
2-[2-trifluorophenylsulfonyl]hydrazide 25
[0183] A solution of 2-phenoxybenzoic acid hydrazide (0.3 M) in
pyridine (0.513 mL) was treated with a solution of the
2-trifluorophenylsulfonyl chloride (1.0 M) in CH.sub.2Cl.sub.2
(0.225 mL). The reaction was placed in a Bohdan Miniblock apparatus
and shaken at 45.degree. C. for 16 hours. The reaction was cooled
to room temperature, teated with polymer-supported polyamine quench
resin (Aldrich, 100 mg), and shaken for 16 hours. The solution was
filtered and concentrated. The residue was purified by preparative
HPLC on a Phenomenex Develofil 28.times.100 mm C-18 column eluting
with a gradient of 10% to 100% CH.sub.3CN/H.sub.2O+3%
n-propanol.
[0184] MS: 437 (M+1 for
C.sub.20H.sub.15F.sub.3N.sub.2O.sub.4S.sub.1); HPLC (Waters
Alliance 2790 column, solvent gradient of 60% to 100%
CH.sub.3CN/H.sub.2O+0.1% formic acid. Purity: 97%.
EXAMPLE 24
[0185] 4-(1,1-Dimethylethyl)-benzoic acid,
2-(phenylsulfonyl)hydrazide. 26
[0186] Step 1: The Preparation of 4-tert-butyl-benzoic acid
hydrazide.
[0187] A solution of 4-tert-butyl benzoic acid (0.2 g, 1.2 mmol in
10 mL of dry THF) cooled to 0.degree. C. and treated with
N-methyl-morpholine (1.6 mL, 14.6 mmol) as well as isobutyl
chloroformate (1.7 mL, 1.3 mmol). Then the mixture was stirred for
10 minutes at 0.degree. C. After hydrazine (0.6 g, 3.6 mmol) was
added to the reaction mixture, it was stirred 3 hours at 25.degree.
C. The reaction was then diluted with ethyl acetate (100 mL),
washed with saturated sodium bicarbonate solution as well as brine,
and concentrated in vacuum. The crude material was chromatographed
on silica gel eluting with hexanes/ethyl acetate=2:1 to give
4-tert-butyl-benzoic acid hydrazide.
[0188] Step 2: The Preparation of 4-(1,1-dimethylethyl)-benzoic
acid, 2-(phenylsulfonyl)hydrazide (Example 24)
[0189] 4-tert-Butyl-benzoic acid hydrazide (0.5 g, 2.60 mmol) was
dissolved in THF (29 mL). N,N-diisopropylethylamine (1.81 mL, 10.4
mmol) was added, and the reaction was cooled to 0.degree. C.
Phenylsulfonyl chloride (0.36 mL, 2.86 mmol) was added followed by
DMAP (5 mg, 0.04 mmol). The reaction was allowed to warm to room
temperature, and the reactants all dissolved. The solution went
from a yellow color to a deep red over the course of 4 hours. The
reaction was then concentrated down and purified by flash
chromatography eluting with 3:1 hexanes/EtOAc. The desired product
was isolated (0.32 g, 37.1%).
[0190] MS: 333.1 (M+1 for C.sub.17H.sub.20N.sub.2O.sub.3S); mp
197-199.degree. C.; TLC: SiO.sub.2 R.sub.f=0.56 (1:1 hexane/EtOAc).
Analysis C.sub.17H.sub.20N.sub.2O.sub.3S: (calc) C, 61.42; H, 6.06;
N, 8.43 (found) C, 61.38; H, 6.09; N, 8.27. IR (KBr, cm.sup.-1)
3213, 2965, 1648, 1403, 1349, 1179. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 1.29 (s, 9H), 7.39-7.55 (m, 8H), 7.92-7.93 (m,
2H), 8.06 (s, 1H).
EXAMPLE 25
[0191] [1,1'-Biphenyl]-4-carboxylic acid,
2-(phenylsulfonyl)hydrazide 27
[0192] Biphenyl-4-carboxylic acid hydrazide (0.5 g, 2.36 mmol) was
suspended in THF (26 mL). N,N-diisopropylethylamine (1.64 mL, 9.44
mmol) was added, and the reaction was cooled to 0.degree. C.
Phenylsulfonyl chloride (0.33 mL, 2.59 mmol) was added followed by
DMAP (5 mg, 0.04 mmol). The reaction was allowed to warm to room
temperature. The solution was orange after it stirred for 1 hour;
DMF (5 mL) was added, and the solution became clear. After 3 hours
of stirring, the solution was diluted with EtOAc (150 mL). It was
then washed with saturated sodium bicarbonate and brine and dried
over Na.sub.2SO.sub.4. The reaction was then concentrated down and
triturated with CH.sub.2Cl.sub.2. The desired product was isolated
(0.19 g, 22.7%).
[0193] MS: 351.1 (M-1 for C.sub.19H.sub.16N.sub.2O.sub.3S); mp
216-219.degree. C. TLC: SiO.sub.2, R.sub.f=0.55 (1:1 hexane/EtOAc).
Analysis C.sub.19H.sub.16N.sub.2O.sub.3S.0.25 H.sub.2O: (calc) C,
63.94; H, 4.66; N, 7.85 (found) C, 63.76; H, 4.64; N, 7.76. HPLC:
(30% H.sub.2O/70% CH.sub.3CN/0.1%TFA), RT=3.731 min. Purity:
94.16%. IR (KBr, cm.sup.-1) 3337, 3035, 2811, 1655, 1340, 1156.
.sup.1H NMR (400 MHz, DMSO) .delta. 7.26-8.00 (m, 14H), 10.01 (s,
1H), 10.71 (s, 1H).
EXAMPLE 26
[0194] 3-Phenoxybenzoic acid, 2-(phenylsulfonyl)hydrazide 28
[0195] Example 26 was synthesized in accordance with the methods of
Example 24 except that 3-phenoxybenzoic acid was used instead of
4-tert-butyl-benzoic acid.
[0196] MS: 369.0 (M+1 for C.sub.19H.sub.16N.sub.2O.sub.4S.sub.1);
amorphous solid; TLC (SiO.sub.2) R.sub.f=0.35 (5%
MeOH/CH.sub.2Cl.sub.2); HPLC (C18 column, 1:1
CH.sub.3CN/H.sub.2O+0.1% TFA) 98.31%, RT=8.863 min. HRMS (calc)
369.0907 (found) 369.0913. IR (KBr, cm.sup.-1): 3309, 3147, 1673,
1165. .sup.1H NMR (DMSO-d.sub.6) .delta. 7.00 (d, 2H, J=8.3 Hz),
7.13-7.19 (m, 3H), 7.38-7.41 (m, 4H), 7.43-7.50 (m, 2H), 7.56-7.59
(m, 1H), 7.76 (d, 2H, J=8.1 Hz), 9.99 (s, 1H), 10.09 (s, 1H).
EXAMPLE 27
[0197] 2-Phenoxybenzoic acid, 2-(phenylsulfonyl)hydrazide 29
[0198] Example 27 was synthesized in accordance with the methods of
Example 24 except that 2-phenoxybenzoic acid was used instead of
4-tert-butyl-benzoic acid.
[0199] MS: 369.0 (M+1 for C.sub.19H.sub.16N.sub.2O.sub.4S.sub.1);
mp 149-150 .degree. C.; TLC (SiO.sub.2) R.sub.f=0.67 (2:1
hexane/EtOAc); HPLC (C18 column, 1:1 CH.sub.3CN/H.sub.2O+0.1% TFA)
99.02%, RT=19.912 min. Analysis
C.sub.19H.sub.16N.sub.2O.sub.4S.sub.1: (calc) C, 61.94; H, 4.38; N,
7.60 (found) C, 61.77; H, 4.33; N, 7.57. IR (KBr, cm.sup.-1): 3368,
3166, 1667, 1479, 1160. .sup.1H NMR (CDCl.sub.3) .delta. 6.80 (d,
1H, J=8.3 Hz), 7.03-7.12 (m, 3H), 7.25-7.27 (m, 1H), 7.36-7.46 (m,
5H), 7.51-7.59 (m, 2H), 7.76-7.83 (m, 3H), 9.39 (d, 1H, J=6.6
Hz).
EXAMPLE 28
[0200] 4-Phenoxybenzoic acid, 2-(phenylsulfonyl)hydrazide 30
[0201] Example 28 was synthesized in accordance with the methods of
Example 24 except that 4-phenoxybenzoic acid was used instead of
4tert-butyl-benzoic acid.
[0202] MS: 369.0 (M+1 for C.sub.19H.sub.16N.sub.2O.sub.4S.sub.1);
mp glassy solid; TLC (SiO.sub.2) R.sub.f=0.70 (1:1 hexane/EtOAc);
HPLC (C18 column, 70:30 CH.sub.3CN/H.sub.2O+0.1% TFA) 94.87%,
RT=3.752 min. HRMS (calc for M+1) 367.0753 (found) 367.0743. IR
(KBr, cm.sup.-1): 3324, 2964, 1723, 1164. .sup.1H NMR
(DMSO-d.sub.6) .delta. 6.92-6.95 (m, 2H), 7.03-7.06 (m, 2H),
7.16-7.20 (m, 1H), 7.38-7.79 (m, 9H), 9.96 (s, 1H), 10.59 (s,
1H).
EXAMPLE 29
[0203] 4-Benzyloxybenzoic acid, 2-phenylsulfonyl)hydrazide 31
[0204] Example 29 was synthesized in accordance with the methods of
Example 24 except that 4-benzyloxybenzoic acid was used instead of
4-tert-butyl-benzoic acid.
[0205] MS: 383.0(M+1 for C.sub.20H.sub.18N.sub.2O.sub.4S.sub.1); mp
159-160.degree. C.; TLC (SiO.sub.2) R.sub.f=0.48 (1:1
hexane/EtOAc); HPLC (C18 column, 1:1 CH.sub.3CN/H.sub.2O+0.1% TFA)
99.85%, RT=11.663 min. IR (KBr, cm.sup.-1): 3403, 3340, 3124, 2941,
1657, 1422, 1165. Analysis C.sub.19H.sub.19N.sub.3O.sub.3S.sub.1:
(calc) C, 62.81; H, 4.74; N, 7.32 (found) C, 62.76; H, 4.78; N,
7.31. .sup.1H NMR (DMSO-d.sub.6) .delta. 5.11 (s, 2H), 7.00 (d, 2H,
J=8.8 Hz), 7.27-7.41 (m, 5H), 7.45-7.49 (m, 2H), 7.55-7.63 (m, 3H),
7.78 (d, 2H, J=7.3 Hz), 9.88 (br, 1H), 10.49 (br, 1H).
EXAMPLE 30
[0206] Benzoic acid, 4-[[(4-methoxyphenyl)sulfonyl]amino]-,
2-(phenylsulfonyl)hydrazide 32
[0207] Example 30 was synthesized in accordance with the methods of
Example 3 except that (4-methoxybenzene)sulfonyl chloride was used
instead of 2,4,6-(trichlorophenyl)sulfonyl chloride (yield
90%).
[0208] MS: 462.1 (M+1 for C.sub.20H.sub.19N.sub.3O.sub.6S.sub.2);
mp 164.3-164.9.degree. C.; HPLC (C18 column, 1:1 MeCN/H.sub.2O=0.1%
TFA) 99.8%, RT=4.2 min. .sup.1H NMR (DMSO-d.sub.6) .delta. 3.13 (S,
3H), 7.04 (d, 4H, J=10.2 Hz), 7.40-7.60 (m, 5H), 7.75 (m, 4H), 9.91
(S, 1H), 10.48 (S, 1H), 10.57 (S, 1H).
EXAMPLE 31
[0209] 4-Iodo-benzoic acid, 2-(phenylsulfonyl)hydrazide 33
[0210] 4-Iodo-benzoic acid, 2-(phenylsulfonyl)hydrazide was
synthesized in accordance with the methods of Example 24, except
that 4iodo-benzoic acid was used instead of 4-nitro-benzoic
acid.
[0211] MS: 402.9 (M+1 for
C.sub.13H.sub.11N.sub.2O.sub.3S.sub.1I.sub.1); mp: 193-195.degree.
C.; TLC (SiO.sub.2) R.sub.f=0.77 (8% MeOH/CH.sub.2Cl.sub.2); HPLC
(C18 column, 1:1 CH.sub.3CN/H.sub.2O+0.1% TFA) 95.71%, RT=7.334
min. HRMS (calc for M+1) 402.9613 (found) 402.9625. IR (KBr,
cm.sup.-1): 3331, 3151, 1645, 1377, 1171. .sup.1H NMR
(DMSO-d.sub.6) .delta. 7.39-7.41 (m, 2H), 7.46-7.52 (m, 2H),
7.54-7.60 (m, 1H), 7.74-7.80 (m, 4H), 10.02, (d, 1H, J=3.6 Hz),
10.72 (d, 1H, J=3.6 Hz).
EXAMPLE 32
[0212] 4-Phenacetylenyl-benzoic acid, 2-(phenylsulfonyl)hydrazide
34
[0213] 4-Iodo-benzoic acid, 2-(phenylsulfonyl)hydrazide (75 mg,
0.19 mmol, Example 31) was dissolved in THF (2.5 mL) and treated
with Et.sub.3N (174 .mu.L, 1.25 mmol), phenylacetylene (41 .mu.L,
0.37 mmol), bis-triphenylphosphine palladium (II) dichloride, and
copper (I) iodide (2.4 mg, 0.013 mmol). The reaction was stirred
overnight at room temperature, then diluted with EtOAc (100 mL),
washed with saturated sodium bicarbonate solution and brine, dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
chromatographed on silica gel eluting with 3% MeOH/CH.sub.2Cl.sub.2
to give 70 mg (97%) of the desired product.
[0214] MS: 377.1 (M+1 for C.sub.21H.sub.16N.sub.2O.sub.3S.sub.1);
mp 175.degree. C. (dec); TLC (SiO.sub.2) R.sub.f=0.50 (4%
MeOH/CH.sub.2Cl.sub.2); HPLC (C18 column, 7:3
CH.sub.3CN/H.sub.2O+0.1% TFA) 94.87%, RT=4.677 min. HRMS (calc for
M+1) 377.0960 (found) 377.0958. IR (KBr, cm.sup.-1): 3324, 3059,
2812, 1658, 1341, 1158. .sup.1H NMR (DMSO-d.sub.6) .delta.
7.39-7.43 (m, 3H), 7.48-7.61 (m, 7H), 7.68 (d, 2H, J=8.3 Hz),
7.79-7.81 (m, 2H), 10.45 (br, 1H), 10.75 (br, 1H).
EXAMPLE 33
[0215] Phenethyl-benzoic acid, 2-(phenylsulfonyl)hydrazide 35
[0216] 4-Phenacetylenyl-benzoic acid, 2-(phenylsulfonyl)hydrazide
(128 mg, 0.34 mmol, Example 32) was dissolved in MeOH (10 mL) and
THF (26 mL), treated with 20% Pd/C (30 mg), and shaken under an
atmosphere of H.sub.2 (50 psi) for 16 hours. The reaction was
filtered and concentrated, then washed with saturated sodium
bicarbonate solution and brine, dried over Na.sub.2SO.sub.4, and
concentrated in vacuum. The residue was chromatographed on silica
gel eluting with 4% MeOH/CH.sub.2Cl.sub.2 to give 125 mg (97%) of
the desired product.
[0217] MS: 381.0 (M+1 for C.sub.21H.sub.20N.sub.2O.sub.3S.sub.1);
mp 158-159.degree. C.; TLC (SiO.sub.2) R.sub.f=0.48 (8%
MeOH/CH.sub.2Cl.sub.2); HPLC (C18 column, 7:3
CH.sub.3CN/H.sub.2O+0.1% TFA) 98.40%, RT=4.544 min. HRMS (calc for
M+1) 381.1273 (found) 381.1270. IR (KBr, cm.sup.-1): 3338, 3134,
1665, 1166. .sup.1H NMR (DMSO-d.sub.6) .delta. 2.81-2.89 (m, 4H),
7.11-7.26 (m, 7H), 7.46-7.60 (m, 5H), 7.77-7.80 (m, 2H), 9.94 (br,
1H), 10.57 (br, 1H).
EXAMPLE 34
[0218] 4-(3-Cyclohexyl-1-propynyl)-benzoic acid,
2-(phenylsulfonyl)hydrazi- de 36
[0219] Example 34 was synthesized in accordance with the methods of
Example 32, except that 3-cyclohexyl-1-propyne was used instead of
phenylacetylene.
[0220] MS: 397 (M+1 for C.sub.22H.sub.24N.sub.2O.sub.3S.sub.1); mp
180-182.degree. C.; TLC (SiO.sub.2) R.sub.f=0.4 (50%
EtOAc/hexanes); HPLC (C18 column, 7:3 CH.sub.3CN/H.sub.2O+0.1% TFA)
98%, RT=9.9 min. .sup.1H NMR (DMSO-d.sub.6) .delta. 0.95-1.24 (m,
5H), 1.43-1.74 (m, 5H), 2.30 (s, 1H), 2.31 (s, 1H), 7.39 (d, 2H,
J=8.6 Hz), 7.46-7.50 (m, 2H), 7.56-7.59 (m, 3H), 7.83 (d, 2H, J=7.3
Hz), 10.00 (s, 1H), 10.70 (s, 1H).
EXAMPLE 35
[0221] 4-(3-Cyclohexyl-3-hydroxy-1-propynyl)-benzoic acid,
2-(phenylsulfonyl)hydrazide 37
[0222] Example 35 was synthesized in accordance with the methods of
Example 32, except that 3-cyclohexyl-3-hydroxy-1-propyne was used
instead of phenylacetylene.
[0223] MS: 399 (M+1 for C.sub.21H.sub.22N.sub.2O.sub.4S.sub.1); mp
96-97.degree. C.; TLC (SiO.sub.2) R.sub.f=0.3 (50% EtOAc/hexanes);
HPLC (C18 column, 7:3 CH.sub.3CN/H.sub.2O+0.1%TFA) 99%, RT=5.0 min.
.sup.1H NMR (DMSO-d.sub.6) .delta. 1.10-1.85 (m, 10H), 5.45 (s,
1H), 7.33-7.50 (m, 4H), 7.52-7.78 (m, 3H), 8.04 (d, 2H, J=7.5 Hz),
10.00 (s, 1H), 10.71 (s, 1H).
EXAMPLE 36
[0224] 4-(3-Cyclohexyl-3-hydroxy-1-propyl)-benzoic acid,
2-(phenylsulfonyl)hydrazide 38
[0225] Example 36 was synthesized in accordance with the methods of
Example 33, except that
4-(3-cyclohexyl-3-hydroxy-1-propynyl)-benzoic acid,
2-(phenylsulfonyl)hydrazide (Example 35) was used instead of
4-phenacetylenyl-benzoic acid, 2-(phenylsulfonyl)hydrazide.
[0226] MS: 403 (M+1 for C.sub.21H.sub.26N.sub.2O.sub.4S.sub.1); mp
158-159.degree. C.; TLC (SiO.sub.2) R.sub.f=0.3 (50%
EtOAc/hexanes); HPLC (C18 column, 7:3 CH.sub.3CN/H.sub.2O+0.1%TFA)
95%, RT=5.0 min. .sup.1H NMR (DMSO-d.sub.6) .delta. 1.10-1.65 (m,
12H), 2.46-2.61 (m, 2H), 4.00 (s, 1H), 7.20 (d, 2 H, J=8.1 Hz),
7.46-7.57 (m, 5H), 7.78 (d, 2H, J=7.3 Hz), 9.93 (s, 1H), 10.56 (s,
1H).
EXAMPLE 37
[0227] 4-(3,3-Dimethyl-1-butynyl)-benzoic acid,
2-(phenylsulfonyl)hydrazid- e 39
[0228] Example 37 was synthesized in accordance with the methods of
Example 32, except that 3,3-dimethylbutyne was used instead of
phenylacetylene.
[0229] MS: 357 (M+1 for C.sub.19H.sub.20N.sub.2O.sub.3S.sub.1); mp
160.degree. C. (dec.); TLC (SiO.sub.2) R.sub.f=0.5 (50%
EtOAc/hexanes); HPLC (C18 column, 7:3 CH.sub.3CN/H.sub.2O+0.1% TFA)
96%, RT=12 min. .sup.1H NMR (DMSO-d6) .delta. 1.23 (s, 9H), 7.49
(d, 2H, J=8.15 Hz), 7.45-7.50 (m, 2H), 7.55-7.60 (m, 3H), 7.77 (d,
2H, J=7.5 Hz), 9.99 (s, 1H), 10.67 (s, 1H).
EXAMPLE 38
[0230] 4-(3,3-Dimethyl-1-butyl)-benzoic acid,
2-(phenylsulfonyl)hydrazide 40
[0231] Example 38 was synthesized in accordance with the methods of
Example 33, except that 4-(3,3-dimethyl-1-butynyl)-benzoic acid,
2-(phenylsulfonyl)hydrazide (Example 37) was used instead of
4-phenacetylenyl-benzoic acid, 2-(phenylsulfonyl)hydrazide.
[0232] MS: 361 (M+1 for C.sub.19H.sub.24N.sub.2O.sub.3S.sub.1); mp
168-169.degree. C.; TLC (SiO.sub.2) R.sub.f=0.7 (50%
EtOAc/hexanes); HPLC (phenol hexal column, 1:1
CH.sub.3CN/H.sub.2O+0.1% TFA) 99%, RT=16.8 min. .sup.1H NMR
(CDCl.sub.3) .delta. 0.95 (s, 9 H), 1.42-1.50 (m, 2H), 2.56-2.62
(m, 2H), 7.21-7.26 (m, 2H), 7.40-7.60 (m, 6H), 7.94 (d, 2H, J=7.8
Hz), 8.13-8.14 (m, 1H).
EXAMPLE 39
In vitro Enzyme Assay: Human Branched Chain Amino Acid
[0233] Aminotransferase cytosolic form (hBCATc) and mitochondrial
form (hBCATm) as well as rat BCAT cytosolic form were assayed at
37.degree. C. in 25 mM phosphate buffer pH 7.8. In addition, 2 mM
DTT and 12.5 mM EDTA were added to the assay mixture. A coupling
enzyme assay was used to monitor the production of glutamate. The
formation of NADH from NAD+ at 340 nM was followed on a 96-well
plate Molecular Devices plate reader. The following components were
used in this coupled assay: 4 mM ADP, 1 mM NAD+, 750 .mu.M
L-Leucine, 500 .mu.M .alpha.-ketoglutarate, 10 .mu.M pyridoxal
phosphate, 1 unit glutamate dehydrogenase, and 1.25 .mu.g of the
appropriate BCAT enzyme. Inhibitions by compounds were assayed by
adding various concentrations to this coupled assay procedure as a
DMSO stock up to a 5% v/v DMSO/buffer ratio. Data is shown in Table
1.
[0234] Enzyme Preparation: humanBCATc and ratBCATc were expressed
in BL21(DE3) cells. A growing culture of cells was induced with 1
mM IPTG at room temperature overnight. These cells were harvested
by centrifugation at 10,000 g for 20 minutes. The supernatant was
discarded, and the cells were stored at -80.degree. C. until
needed. The protein was purified by the following method. The cell
paste was resuspended in extraction buffer (0.1 M phosphate, pH
8.0, with 0.01 M Tris-HCL and 5 mM TCEP and then lysed on a French
Press at 1000 psi. The lysate was centrifuged at 10,000 g for 15
minutes, and the pellet was discarded. The supernatant was loaded
onto a Hitrap Chelating column previously charged with 0.1 M
NiSO.sub.4 and washed with 3 to 4 column volumes of extraction
buffer. The column was then washed with 0.01 Tris, pH 7.5, 10%
glycerol, 150 mM NaCl, 5 mM TCEP for two column volumes. The column
was then further washed with two column volumes of 0.1 M phosphate,
pH 6.0, 0.01 M Tris, 10% glycerol, and 750 mM NaCl. Finally, the
column was washed with two column volumes of the same buffer plus
50 mM imidazole. The BCAT enzyme was then eluted with the same
buffer but with 350 mM imidazole. The eluant was then dialyzed
overnight against 10 mM phosphate buffer pH 8.0, 10% glycerol, and
5 mM TCEP. The dialyzed protein was then loaded onto a Q-sepharose
column and eluted using a salt gradient. Active fractions were
collected and further purified on a Superose-12 column to yield
pure BCAT protein.
1TABLE 1 In vitro Data Example No. rrBCATc (.mu.M) rhBCATc (.mu.M)
1 40 2 28.1 60 3 0.7 4 6 5 10 6 3 7 8 7 9 3.8 10 13 11 7.5 12 4.6
13 14 4.8 15 4.9 16 4.4 27 17 2.3 18 1.2 9.5 19 2.8 28 20 5.9 35 21
2.9 28 22 110 23 0.43 17 24 1.2 57 25 1.5 16 26 2.2 34 27 50 28 1.5
37 29 48 26 30 2.8 31 32 1.3 27.6 33 0.25 14.3 34 32.3 35 2.2 43.5
36 0.25 15.4 37 4.3 34.8 38 0.25 9.5
* * * * *