U.S. patent application number 14/138592 was filed with the patent office on 2014-07-03 for dual mechanism inhibitors for the treatment of disease.
This patent application is currently assigned to AERIE PHARMACEUTICALS, INC.. The applicant listed for this patent is AERIE PHARMACEUTICALS, INC.. Invention is credited to Mitchell A. deLong, Susan M. Royalty, Jill Marie Sturdivant.
Application Number | 20140187617 14/138592 |
Document ID | / |
Family ID | 43017737 |
Filed Date | 2014-07-03 |
United States Patent
Application |
20140187617 |
Kind Code |
A1 |
deLong; Mitchell A. ; et
al. |
July 3, 2014 |
DUAL MECHANISM INHIBITORS FOR THE TREATMENT OF DISEASE
Abstract
Provided are compounds that are inhibitors of both rho kinase
and of a monoamine transporter (MAT) act to improve the disease
state or condition. Further provided are compositions comprising
the compounds. Further provided are methods for treating diseases
or conditions, the methods comprising administering compounds
according to the invention. One such disease may be glaucoma for
which, among other beneficial effects, a marked reduction in
intraocular pressure (IOP) may be achieved.
Inventors: |
deLong; Mitchell A.; (Chapel
Hill, NC) ; Sturdivant; Jill Marie; (Chapel Hill,
NC) ; Royalty; Susan M.; (Davis, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AERIE PHARMACEUTICALS, INC. |
Research Triangle Park |
NC |
US |
|
|
Assignee: |
AERIE PHARMACEUTICALS, INC.
Research Triangle Park
NC
|
Family ID: |
43017737 |
Appl. No.: |
14/138592 |
Filed: |
December 23, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13768594 |
Feb 15, 2013 |
8716310 |
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14138592 |
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12694965 |
Jan 27, 2010 |
8394826 |
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13768594 |
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61174672 |
May 1, 2009 |
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Current U.S.
Class: |
514/448 ;
435/375; 514/530; 514/534; 514/543; 514/546; 549/71; 560/107;
560/122; 560/251; 560/75 |
Current CPC
Class: |
A61P 43/00 20180101;
A61P 1/18 20180101; C07D 401/12 20130101; C07D 413/12 20130101;
C07D 417/12 20130101; C07C 237/20 20130101; C07D 217/22 20130101;
A61P 17/00 20180101; A61P 29/00 20180101; C07D 217/24 20130101;
A61P 3/04 20180101; A61P 11/02 20180101; A61P 19/02 20180101; A61P
1/04 20180101; A61P 27/04 20180101; A61P 17/14 20180101; C07D
217/04 20130101; C07C 2601/08 20170501; C07D 217/00 20130101; A61P
19/08 20180101; C07D 333/40 20130101; C07D 409/12 20130101; A61P
1/16 20180101; A61P 27/06 20180101; A61P 13/12 20180101; C07D
333/38 20130101; A61P 1/00 20180101; C07C 2601/14 20170501; C07D
405/12 20130101; A61P 27/00 20180101; A61P 35/00 20180101; A61P
13/10 20180101; C07C 237/42 20130101; C07D 217/02 20130101; A61P
9/00 20180101; A61P 27/02 20180101; A61P 15/18 20180101; A61P 9/10
20180101; A61P 9/12 20180101; A61P 11/00 20180101 |
Class at
Publication: |
514/448 ;
560/107; 514/534; 560/251; 514/546; 560/122; 514/530; 549/71;
560/75; 514/543; 435/375 |
International
Class: |
C07C 237/42 20060101
C07C237/42; C07D 333/40 20060101 C07D333/40 |
Claims
1. A compound according to Formula II: ##STR00231## and salts
thereof, wherein R.sup.1, R.sup.2, and R.sup.3 are independently
hydrogen, C.sub.1-C.sub.4 alkyl, aryl, C.sub.1-C.sub.4 alkyl aryl,
C.sub.1-C.sub.4 alkyl heteroaryl, C.sub.1-C.sub.4 alkyl
heterocyclyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.1-C.sub.4 carbonyl, C.sub.1-C.sub.4 carbonylamino,
C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 sulfonyl, C.sub.1-C.sub.4
sulfonylamino, C.sub.1-C.sub.4 thioalkyl, C.sub.1-C.sub.4 carboxyl,
or form a ring with each other or with A; wherein A is
C.sub.1-C.sub.4 alkyl, or forms a ring structure with R.sup.1,
R.sup.2, or R.sup.3; wherein B is hydrogen, an aryl group, a
heteroaryl group, a cycloalkyl group, a heterocycloalkyl group,
C.sub.1-C.sub.22 alkyl, C.sub.1-C.sub.22 alkyl aryl,
C.sub.1-C.sub.22 alkyl heteroaryl, C.sub.2-C.sub.22 alkenyl,
C.sub.2-C.sub.22 alkynyl, C.sub.1-C.sub.22 carbonyl,
C.sub.1-C.sub.22 carbonylamino, C.sub.1-C.sub.22 alkoxy,
C.sub.1-C.sub.22 sulfonyl, C.sub.1-C.sub.22 sulfonylamino,
C.sub.1-C.sub.22 thioalkyl, or C.sub.1-C.sub.22 carboxyl; wherein
X.sup.1, X.sup.2, and X.sup.3 are, independently, hydrogen,
hydroxyl, halogen, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 alkynyl, amino, aminocarbonyl, nitro, cyano,
C.sub.1-C.sub.4 carbonyl, C.sub.1-C.sub.4 carbonylamino,
C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 sulfonyl, C.sub.1-C.sub.4
sulfonylamino, C.sub.1-C.sub.4 thioalkyl, or C.sub.1-C.sub.4
carboxyl; wherein the double circle ##STR00232## indicates an
aromatic or heteroaromatic ring; and wherein each Z is
independently a bond, C.sub.1-C.sub.4 alkyl, heteroalkyl, or an O
atom.
2. A compound according to claim 1, wherein R.sup.1 is hydrogen or
C.sub.1-C.sub.4 alkyl; B is C.sub.1-C.sub.22 carbonyl; X.sub.1,
X.sub.2, and X.sub.3 are hydrogen; the double circle ##STR00233##
indicates an aromatic or heteroaromatic ring; and Z is a bond,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl, or an O
atom.
3. A compound according to claim 1, wherein Z is a bond or
--CH.sub.2--.
4. A compound according to claim 1, wherein X.sup.1, X.sup.2 and
X.sup.3 are each hydrogen.
5. A compound according to claim 1, wherein A is
--CH.sub.2CH.sub.2--.
6. A compound according to claim 1, wherein R.sup.1 and R.sup.2 are
independently hydrogen or methyl.
7. A compound according to claim 1, wherein R.sup.3 is hydrogen or
C.sub.1-C.sub.4 alkyl.
8. A pharmaceutical composition comprising a compound according to
claim 1 and a pharmaceutically acceptable carrier.
9. A method for treating a disease in a subject comprising:
administering to a subject an effective amount of a compound
according to claim 1; wherein the disease comprises eye
disease.
10. The method of claim 9, wherein the eye disease comprises
glaucoma or a neurodegenerative eye disease.
11. A method of modulating kinase activity comprising: contacting a
cell with a compound according to claim 1 in an amount effective to
modulate kinase activity.
12. The method of claim 11, wherein the cell is in a subject.
13. A method of reducing intraocular pressure comprising:
contacting a cell with a compound according to claim 1 in an amount
effective to reduce intraocular pressure.
14. The method of claim 13, wherein the cell is in a subject.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 13/768,594 filed Feb. 15, 2013 which is a
divisional of U.S. patent application Ser. No. 12/694,965, filed
Jan. 27, 2010, which issued as U.S. Pat. No. 8,394,826 on Mar. 12,
2013, which claims priority to U.S. Provisional Patent Application
No. 61/174,672, filed May 1, 2009, each of which are incorporated
herein by reference in their entireties.
FIELD OF THE INVENTION
[0002] The present invention relates to substituted isoquinoline
amide compounds and substituted benzamide compounds that affect the
function of kinases and the function of transporters in a cell and
that are useful as therapeutic agents or in conjunction with
therapeutic agents. In particular, these compounds are useful in
the treatment of diseases and disorders of the eye, such as
glaucoma, of the respiratory system, of the cardiovascular system,
and for diseases characterized by abnormal growth, such as
cancers.
BACKGROUND
[0003] A wide variety of hormones, neurotransmitters, and other
biologically active substances control, regulate, or adjust the
functions of the body via interaction with specific cellular
receptors. Many of these receptors mediate the transmission of
intracellular signals by activating guanine nucleotide-binding
proteins (G proteins) to which the receptor is coupled. Such
receptors are generically referred to as G-protein coupled
receptors (GPCRs) and include, among others, adrenergic receptors,
opioid receptors, cannabinoid receptors, and prostaglandin
receptors. The biological effects of activating these receptors are
not direct but are mediated by a host of intracellular proteins.
The importance of these secondary proteins has only recently been
recognized and investigated as intervention points in disease
states. One of the most important classes of downstream effectors
is the "kinase" class.
[0004] Various kinases play important roles in the regulation of
many physiological functions. For example, kinases have been
implicated in numerous disease states including, but not limited
to, cardiac disorders such as angina pectoris, essential
hypertension, myocardial infarction, supraventricular and
ventricular arrhythmias, congestive heart failure, and
atherosclerosis, respiratory disorders such as asthma, chronic
bronchitis, bronchospasm, emphysema, airway obstruction, rhinitis,
and seasonal allergies, inflammation, rheumatoid arthritis, renal
failure, and diabetes. Other conditions include chronic
inflammatory bowel disease, glaucoma, hypergastrinemia,
gastrointestinal indications such as acid/peptic disorder, erosive
esophagitis, gastrointestinal hypersecretion, mastocytosis,
gastrointestinal reflux, peptic ulcer, pain, obesity, bulimia
nervosa, depression, obsessive-compulsive disorder, organ
malformations (e.g., cardiac malformations), neurodegenerative
diseases such as Parkinson's Disease and Alzheimer's Disease,
multiple sclerosis, Epstein-Barr infection, and cancer (Nature
Reviews Drug Discovery 2002, 1: 493-502). In other disease states,
the role of kinases is only now becoming clear.
[0005] The success of the tyrosine-kinase inhibitor STI571
(Gleevec) in the treatment of chronic myelogenous leukemia (Nature
Reviews Drug Discovery 2003, 2: 296-313) has spurred considerable
efforts to develop other kinase inhibitors for the treatment of a
wide range of other cancers (Nature Reviews Cancer 2003, 3:
650-665). Seven additional kinase inhibitor drugs have since been
brought to market, establishing kinase inhibitors as an important
new drug class. Currently more than 100 protein kinase inhibitors
are in clinical development (Kinase Inhibitor Drugs 2009, Wiley
Press).
[0006] In view of the role that kinases have in many disease
states, there is an urgent and continuing need for small molecule
ligands that inhibit or modulate the activity of kinases. Without
wishing to be bound by theory, it is thought that modulation of the
activity of kinases, including rho kinase (ROCK), by the compounds
of the present invention is, in part, responsible for their
beneficial effects.
[0007] An additional area of fruitful research in medicine is the
study of monoamine transporters and the benefits of inhibition
thereof. Monoamine transporters (MAT) are structures in cell
membranes that transport monoamine-containing neurotransmitters
into or out of cells. There are several distinct monoamine
transporters, or MATs: the dopamine transporter (DAT), the
norepinephrine transporter (NET), and the serotonin transporter
(SERT). DAT, NET, and SERT are structurally-related, and each
contains a structure of 12 trans-membrane helices. Modern
antidepressants are thought to work by enhancing serotonergic,
noradrenergic, or dopaminergic neurotransmission by binding to
their respective transporter, and thereby inhibiting
neurotransmitter reuptake and effectively raising the concentration
of the neurotransmitter in synapses. Examples of drugs that are
thought to operate by this mechanism include fluoxetine, a
selective SERT inhibitor; reboxetine, a norepinephrine (NET)
inhibitor; and bupropion, which inhibits both the NET and DAT (He,
R. et al. J. Med. Chem. 2005, 48: 7970-9; Blough, B. E. et al. J.
Med. Chem. 2002, 45: 4029-37; Blough, B. E., et al. J. Med. Chem.
1996, 39: 4027-35; Torres, G. E., et al. Nat. Rev. Neurosci. 2003,
4: 13-25).
[0008] Glaucoma causes deterioration of the eye's optic nerve, the
nerve bundles that carry images from the ganglion cells of the eye
to the brain. Increased IOP is a hallmark of the most common forms
of glaucoma, and this increased intraocular pressure likely damages
the optic nerve and ganglion cells through multiple mechanisms.
Current glaucoma medications act by reducing intraocular pressure,
either by slowing the flow of aqueous humor into the eye or by
improving the drainage of this fluid from the eye. Inhibitors of
rho kinase have been shown to reduce IOP in rabbits and monkeys by
increasing aqueous humor drainage through the trabecular meshwork
(Tian and Kaufman, Arch Ophthalmol 2004, 122: 1171-1178; Tokushige
et al., IOVS 2007, 48(7): 3216-3222). Several lines of experimental
evidence indicate that modulating the activity of rho kinase within
the aqueous humor outflow pathway could be beneficial for the
treatment of patients with glaucoma (Honjo et al., IOVS 2001; 42:
137-144; Waki et al., Curr Eye Res 2001; 22: 470-474; Rao et al.,
IOVS 2001; 42: 1029-1037). Considerable evidence suggests that the
sympathetic nervous system plays a significant but complex role in
the regulation of IOP (Nathanson, Proc. Natl. Acad. Sci. USA 1980;
77(12):7420-7424). Sympathetic nerve fibers innervate the ciliary
process and trabecular meshwork (Ehinger, Acta Univ. Lund Sect 2
1964; 20:3-23; Sears, 1975, Handbook of Physiology, Endocrinology
VI. Eds Astwood E & Greep R: 553-590) and both sympathetic
stimulation and locally applied .beta.-adrenergic agonists such as
epinephrine decrease IOP (Sears, 1975, ibid; Dayson et al., J.
Physiol. (London) 1951; 113:389-397).
SUMMARY OF THE INVENTION
[0009] In one aspect, the may invention provide compounds according
to Formulas I, II, III, IV, V, or VI, as describe below.
[0010] In other aspects, the invention may provide pharmaceutical
compositions, comprising a compound according to Formula I, II,
III, IV, V, or VI as described below, and a carrier.
[0011] In further aspects, the invention may provide methods of
treating a disease in a subject, the method comprising
administering to a subject an effective amount of a compound
according to Formula I, II, III, IV, V, or VI as described below.
The disease may be selected from the group consisting of eye
disease including glaucoma and retinal diseases such as Wet AMD Dry
AMD (inflammation) and DME, bone disorder including osteoporosis,
vascular disease including cerebral vasospasm, coronary vasospasm,
hypertension, pulmonary hypertension, sudden death syndrome,
angina, myocardial infarction, restenosis, stroke, hypertensive
vascular disease, heart failure, cardiac allograft vasculopathy,
vein graft disease, pulmonary disease including chronic obstructive
pulmonary disease (COPD) and asthma, neurological disorder
including spinal cord injury, Alzheimer's disease, multiple
sclerosis, depression, attention deficit-hyperactivity disorder and
neuropathic pain, neovascular disorders and cancer, obesity, and
erectile dysfunction.
[0012] In further aspects, the invention may provide methods of
modulating kinase activity, the methods comprising contacting a
cell with a compound according to Formula I, II, III, IV, V, or VI
as described below, in an amount effective to modulate kinase
activity.
[0013] In further aspects, the invention may provide methods of
reducing intraocular pressure, the methods comprising contacting a
cell with a compound according to Formula I, II, III, IV, V or VI
as described below, in an amount effective to reduce intraocular
pressure.
BRIEF DESCRIPTION OF THE DRAWINGS
[0014] FIG. 1 is a scheme for the synthesis of compounds, including
E1-E8.
[0015] FIG. 2 is a scheme for the synthesis of compounds, including
E457-E466.
[0016] FIG. 3 is a scheme for the synthesis of benzamidines,
E467-E476.
[0017] FIG. 4 is a scheme for the synthesis of para-aminobenzamide
precursors, E477-E478, for the synthesis scheme of FIG. 3.
[0018] FIG. 5 is a scheme for the synthesis of para-aminobenzamide
precursors, E479-E481, for the synthesis scheme of FIG. 3.
[0019] FIG. 6 is a scheme for the synthesis of compounds, including
E8-E12.
[0020] FIG. 7 is a scheme for the synthesis of compounds, including
E132-E139.
[0021] FIG. 8 is a scheme for the synthesis of compounds, including
E140-E143.
[0022] FIG. 9 is a scheme for the synthesis of compounds, including
E145-E148.
[0023] FIG. 10 is a scheme for the synthesis of compounds,
including E197-S and E197-R.
[0024] FIG. 11 is a scheme for the synthesis of compounds,
including E199-E203.
[0025] FIG. 12 is a scheme for the synthesis of compounds,
including E204-E206.
[0026] FIG. 13 is a scheme for the synthesis of compounds,
including E231-E241.
[0027] FIG. 14 is a scheme for the synthesis of compounds,
including E249-253.
[0028] FIG. 15 is a scheme for the synthesis of compounds,
including E275-E278.
[0029] FIG. 16 is a scheme for the synthesis of compounds,
including E289-E290.
[0030] FIG. 17 is a scheme for the synthesis of compounds,
including E300-E308.
[0031] FIG. 18 is a scheme for the synthesis of compounds,
including E319-E325.
[0032] FIG. 19 is a scheme for the synthesis of compounds,
including E371-E377.
[0033] FIG. 20 is a scheme for the synthesis of compounds,
including E398-E404.
[0034] FIG. 21 is a general scheme for the synthesis of compounds,
including compounds E429-E433.
DETAILED DESCRIPTION
[0035] Publications and patents are referred to throughout this
disclosure. All U.S. patents cited herein are hereby incorporated
by reference. All percentages, ratios, and proportions used herein
are percent by weight unless otherwise specified.
[0036] Amino isoquinolyl amides and amino benzamidyl amides are
provided. In some aspects, the compositions and methods for
treating diseases and conditions wherein compounds that may be
inhibitors of both rho kinase and of a monoamine transporter (MAT)
act to improve the disease state or condition. One such disease may
be glaucoma for which, among other beneficial effects, a marked
reduction in intraocular pressure (IOP) may be achieved.
DEFINITIONS
[0037] "Alkyl" refers to a saturated aliphatic hydrocarbon moiety
including straight chain and branched chain groups. "Alkyl" may be
exemplified by groups such as methyl, ethyl, n-propyl, isopropyl,
n-butyl and the like. Alkyl groups may be substituted or
unsubstituted. Substituents may also be themselves substituted.
[0038] When substituted, the substituent group is preferably but
not limited to C.sub.1-C.sub.4 alkyl, aryl, carbocyclyl,
heterocarbocyclyl, heteroaryl, amino, cyano, halogen, alkoxy or
hydroxyl. "C.sub.1-C.sub.4 alkyl" refers to alkyl groups containing
one to four carbon atoms.
[0039] "Alkenyl" refers to an unsaturated aliphatic hydrocarbon
moiety including straight chain and branched chain groups. Alkenyl
moieties must contain at least one alkene. "Alkenyl" may be
exemplified by groups such as ethenyl, n-propenyl, isopropenyl,
n-butenyl and the like. Alkenyl groups may be substituted or
unsubstituted. Substituents may also be themselves substituted.
When substituted, the substituent group is preferably alkyl, aryl,
carbocyclyl, heterocarbocyclyl, heteroaryl, halogen or alkoxy.
Substituents may also be themselves substituted. Substituents may
be placed on the alkene itself and also on the adjacent member
atoms or the alkynyl moiety. "C.sub.2-C.sub.4 alkenyl" refers to
alkenyl groups containing two to four carbon atoms.
[0040] "Alkynyl" refers to an unsaturated aliphatic hydrocarbon
moiety including straight chain and branched chain groups. Alkynyl
moieties must contain at least one alkyne. "Alkynyl" may be
exemplified by groups such as ethynyl, propynyl, n-butynyl and the
like. Alkynyl groups may be substituted or unsubstituted. When
substituted, the substituent group is preferably alkyl, aryl,
carbocyclyl, heterocarbocyclyl, heteroaryl, amino, cyano, halogen,
alkoxyl or hydroxyl. Substituents may also be themselves
substituted. When substituted, substituents are not on the alkyne
itself but on the adjacent member atoms of the alkynyl moiety.
"C.sub.2-C.sub.4 alkynyl" refers to alkynyl groups containing two
to four carbon atoms.
[0041] "Acyl" or "carbonyl" refers to the moiety --C(O)R wherein R
is alkyl, alkenyl, alkynyl, aryl, heteroaryl, carbocyclic,
heterocarbocyclic, C.sub.1-C.sub.4 alkyl aryl, or C.sub.1-C.sub.4
alkyl heteroaryl. C.sub.1-C.sub.4 alkylcarbonyl refers to a group
wherein the carbonyl moiety is preceded by an alkyl chain of 1-4
carbon atoms.
[0042] "Alkoxy" refers to the moiety --O--R wherein R is acyl,
alkyl alkenyl, alkyl alkynyl, aryl, carbocyclic, heterocarbocyclic,
heteroaryl, C.sub.1-C.sub.4 alkyl aryl, or C.sub.1-C.sub.4 alkyl
heteroaryl.
[0043] "Amino" refers to the moiety --NR'R' wherein each R' is,
independently, hydrogen, amino, hydroxyl, alkoxyl, alkyl, aryl,
cycloalkyl, heterocycloalkyl, heteroaryl, C.sub.1-C.sub.4 alkyl
aryl, or C.sub.1-C.sub.4 alkyl heteroaryl. The two R' groups may
themselves be linked to form a ring. The R' groups may themselves
be further substituted, in which case the group also known as
guanidinyl is specifically contemplated under the term "amino".
[0044] "Aryl" or "aromatic ring" refers to an aromatic carbocyclic
moiety. "Aryl" may be exemplified by phenyl. The aryl group may be
substituted or unsubstituted. Substituents may also be themselves
substituted. When substituted, the substituent group is preferably
but not limited to alkoxyl, heteroaryl, acyl, carboxyl,
carbonylamino, nitro, amino, cyano, halogen, or hydroxyl.
[0045] "Carboxyl" refers to the group --C(.dbd.O)O--R wherein R is
chosen from hydrogen, alkyl, aryl, cycloalkyl, heterocycloalkyl,
heteroaryl, C.sub.1-C.sub.4 alkyl aryl, or C.sub.1-C.sub.4 alkyl
heteroaryl.
[0046] "Carbonyl" refers to the group --C(O)R wherein each R is,
independently, hydrogen, alkyl, aryl, cycloalkyl, heterocycloalkyl,
heteroaryl, C.sub.1-C.sub.4 alkyl aryl, or C.sub.1-C.sub.4 alkyl
heteroaryl.
[0047] "Carbonylamino" refers to the group --C(O)NR'R' wherein each
R' is, independently, hydrogen, alkyl, aryl, cycloalkyl,
heterocycloalkyl, heteroaryl, C.sub.1-C.sub.4 alkyl aryl, or
C.sub.1-C.sub.4 alkyl heteroaryl. The two R' groups may themselves
be linked to form a ring. Carbonylamino is also known as an amide
linkage.
[0048] "C.sub.1-C.sub.4 alkyl aryl" refers to C.sub.1-C.sub.4 alkyl
groups having an aryl substituent such that the aryl substituent is
bonded through an alkyl group. "C.sub.1-C.sub.4 alkyl aryl" may be
exemplified by benzyl and phenethyl.
[0049] "C.sub.1-C.sub.4 alkyl heteroaryl" refers to C.sub.1-C.sub.4
alkyl groups having a heteroaryl substituent such that the
heteroaryl substituent is bonded through an alkyl group.
[0050] "Carbocyclic group" or "cycloalkyl" means a monovalent
saturated or unsaturated hydrocarbon ring. Carbocyclic groups are
monocyclic, or are fused, spiro, or bridged bicyclic ring systems.
Monocyclic carbocyclic groups contain 3 to 10 carbon atoms,
preferably 4 to 7 carbon atoms, and more preferably 5 to 6 carbon
atoms in the ring. Bicyclic carbocyclic groups contain 8 to 12
carbon atoms, preferably 9 to 10 carbon atoms in the ring.
Carbocyclic groups may be substituted or unsubstituted.
Substituents may also be themselves substituted. Preferred
carbocyclic groups include cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cyclohexenyl, and cycloheptyl. More preferred
carbocyclic groups include cyclopentyl and cyclohexyl. The most
preferred carbocyclic group is cyclohexyl. Carbocyclic groups are
not aromatic.
[0051] "Halogen" refers to fluoro, chloro, bromo, or iodo moieties.
Preferably, the halogen is fluoro, chloro, or bromo.
[0052] "Heteroaryl" or "heteroaromatic ring" refers to a monocyclic
or bicyclic aromatic carbocyclic radical having one or more
heteroatoms in the carbocyclic ring. Heteroaryl may be substituted
or unsubstituted. When substituted, the substituents may themselves
be substituted. Non-limiting examples of substituents may include
aryl, C.sub.1-C.sub.4 alkylaryl, amino, halogen, hydroxy, cyano,
nitro, carboxyl, carbonylamino, or C.sub.1-C.sub.4 alkyl. Preferred
heteroaromatic groups include tetrazoyl, triazolyl, thienyl,
thiazolyl, purinyl, pyrimidyl, pyridyl, and furanyl. More preferred
heteroaromatic groups include benzothiofuranyl, thienyl, furanyl,
tetrazoyl, triazolyl, and pyridyl.
[0053] "Heteroatom" means an atom other than carbon in the ring of
a heterocyclic group or a heteroaromatic group or the chain of a
heterogeneous group. Preferably, heteroatoms are selected from the
group consisting of nitrogen, sulfur, and oxygen atoms. Groups
containing more than one heteroatom may contain different
heteroatoms.
[0054] "Heterocarbocyclic group" or "heterocycloalkyl" or
"heterocyclic" means a monovalent saturated or unsaturated
hydrocarbon ring containing at least one heteroatom.
Heterocarbocyclic groups are monocyclic, or are fused, spiro, or
bridged bicyclic ring systems. Monocyclic heterocarbocyclic groups
contain 3 to 10 carbon atoms, preferably 4 to 7 carbon atoms, and
more preferably 5 to 6 carbon atoms in the ring. Bicyclic
heterocarbocyclic groups contain 8 to 12 carbon atoms, preferably 9
to 10 carbon atoms in the ring. Heterocarbocyclic groups may be
substituted or unsubstituted. Substituents may also be themselves
substituted. Preferred heterocarbocyclic groups include epoxy,
tetrahydrofuranyl, azacyclopentyl, azacyclohexyl, piperidyl, and
homopiperidyl. More preferred heterocarbocyclic groups include
piperidyl and homopiperidyl. The most preferred heterocarbocyclic
group is piperidyl. Heterocarbocyclic groups are not aromatic.
[0055] "Hydroxy" or "hydroxyl" means a chemical entity that
consists of --OH. Alcohols contain hydroxy groups. Hydroxy groups
may be free or protected. An alternative name for hydroxyl is
alcohol.
[0056] "Linker" means a linear chain of n member atoms where n is
an integer of from about 1 to about 4 member atoms.
[0057] "Member atom" means a carbon, nitrogen, oxygen, or sulfur
atom. Member atoms may be substituted up to their normal valence.
If substitution is not specified the substituents required for
valency are hydrogen.
[0058] "Ring" means a collection of member atoms that are cyclic.
Rings may be carbocyclic, aromatic, heterocyclic, or
heteroaromatic, and may be substituted or unsubstituted, and may be
saturated or unsaturated. Ring junctions with the main chain may be
fused or spirocyclic. Rings may be monocyclic or bicyclic. Rings
contain at least 3 member atoms and at most 10 member atoms.
Monocyclic rings may contain 3 to 7 member atoms and bicyclic rings
may contain from 8 to 12 member atoms. Bicyclic rings themselves
may be fused or spirocyclic.
[0059] "Thioalkyl" refers to the group --S-alkyl.
[0060] "Sulfonyl" refers to the --S(O).sub.2R' group wherein R' is
alkoxy, alkyl, aryl, carbocyclic, heterocarbocyclic, heteroaryl,
C.sub.1-C.sub.4 alkyl aryl, or C.sub.1-C.sub.4 alkyl
heteroaryl.
[0061] "Sulfonylamino" refers to the --S(O).sub.2NR'R' group
wherein each R' is independently alkyl, aryl, heteroaryl,
C.sub.1-C.sub.4 alkyl aryl, or C.sub.1-C.sub.4 alkyl
heteroaryl.
[0062] "Pharmaceutically acceptable carrier" means a carrier that
is useful for the preparation of a pharmaceutical composition that
is generally compatible with the other ingredients of the
composition, not deleterious to the recipient, and neither
biologically nor otherwise undesirable. "A pharmaceutically
acceptable carrier" includes both one and more than one carrier.
Embodiments include carriers for topical, ocular, parenteral,
intravenous, intraperitoneal intramuscular, sublingual, nasal, and
oral administration. "Pharmaceutically acceptable carrier" also
includes agents for preparation of aqueous dispersions and sterile
powders for injection or dispersions.
[0063] "Excipient" as used herein includes physiologically
compatible additives useful in preparation of a pharmaceutical
composition. Examples of pharmaceutically acceptable carriers and
excipients can for example be found in Remington Pharmaceutical
Science, 16.sup.th Ed.
[0064] "Effective amount" as used herein refers to a dosage of the
compounds or compositions effective for influencing, reducing, or
inhibiting the activity of or preventing activation of a kinase or
a transporter protein. This term as used herein may also refer to
an amount effective at bringing about a desired in vivo effect in
an animal, preferably, a human, such as reduction in intraocular
pressure.
[0065] "Administering" as used herein refers to administration of
the compounds as needed to achieve the desired effect.
[0066] "Eye disease" as used herein includes, but is not limited
to, glaucoma, allergy, cancers of the eye, neurodegenerative
diseases of the eye, and dry eye.
[0067] The term "disease or condition associated with kinase
activity" is used to mean a disease or condition treatable, in
whole or in part, by inhibition of one or more kinases. The
diseases or conditions may include, but are not limited to, eye
disease including glaucoma and retinal diseases such as Wet AMD Dry
AMD (inflammation) and DME, ocular hypertension, bone disorder
including osteoporosis, vascular disease including cerebral
vasospasm, cardiovascular diseases, coronary vasospasm,
hypertension, pulmonary hypertension, sudden death syndrome,
angina, myocardial infarction, restenosis, stroke, hypertensive
vascular disease, heart failure, cardiac allograft vasculopathy,
vein graft disease, pulmonary disease including chronic obstructive
pulmonary disease (COPD) and asthma, neurological disorder
including spinal cord injury, Alzheimer's disease, multiple
sclerosis, depression, attention deficit-hyperactivity disorder and
neuropathic pain, neovascular disorders and cancer, obesity, and
erectile dysfunction.
[0068] The term "controlling the disease or condition" is used to
mean changing the activity of one or more kinases to affect the
disease or condition.
[0069] The term "contacting a cell" is used to mean contacting a
cell in vitro or in vivo (i.e. in a subject, such as a mammal,
including humans, cats, and dogs).
Compounds
[0070] In a first aspect of the invention, provided are
isoquinoline amide compounds. Compounds according to the inventions
include those according to Formula I:
##STR00001##
wherein R.sup.1, R.sup.2, and R.sup.3 are independently hydrogen,
C.sub.1-C.sub.4 alkyl, aryl, C.sub.1-C.sub.4 alkyl aryl,
C.sub.1-C.sub.4 alkyl heteroaryl, C.sub.1-C.sub.4 alkyl
heterocyclyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.1-C.sub.4 carbonyl, C.sub.1-C.sub.4 carbonylamino,
C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 sulfonyl, C.sub.1-C.sub.4
sulfonylamino, C.sub.1-C.sub.4 thioalkyl, C.sub.1-C.sub.4 carboxyl,
or form a ring with each other or with A; wherein A is
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkyl aryl, C.sub.1-C.sub.4
alkyl heteroaryl, or forms a ring structure with R.sup.1, R.sup.2
or R.sup.3; wherein B is hydrogen, an aryl group, a heteroaryl
group, a cycloalkyl group, a heterocycloalkyl group,
C.sub.1-C.sub.22 alkyl, C.sub.1-C.sub.22 alkyl aryl,
C.sub.1-C.sub.22 alkyl heteroaryl, C.sub.2-C.sub.22 alkenyl,
C.sub.2-C.sub.22 alkynyl, C.sub.1-C.sub.22 carbonyl,
C.sub.1-C.sub.22 carbonylamino, C.sub.1-C.sub.22 alkoxy,
C.sub.1-C.sub.22 sulfonyl, C.sub.1-C.sub.22 sulfonylamino,
C.sub.1-C.sub.22 thioalkyl, or C.sub.1-C.sub.22 carboxyl, the
stereocenters present, if any, being either `R` or `S` in
configuration independently; wherein X.sup.1, X.sup.2, and X.sup.3
are, independently, hydrogen, hydroxyl, halogen, C.sub.1-C.sub.4
alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 alkynyl, amino,
aminocarbonyl, nitro, cyano, C.sub.1-C.sub.4 carbonyl,
C.sub.1-C.sub.4 carbonylamino, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.4 sulfonyl, C.sub.1-C.sub.4 sulfonylamino,
C.sub.1-C.sub.4 thioalkyl, or C.sub.1-C.sub.4 carboxyl; wherein the
double circle
##STR00002##
indicates an aromatic or heteroaromatic ring; and wherein each Z is
independently a bond, C.sub.1-C.sub.4 alkyl, heteroalkyl, or an O
atom.
[0071] In Formula I, any and all of the stereocenters may be
independently either `R` or `S` in configuration.
[0072] Certain embodiments of Formula I include compounds of
Formula III:
##STR00003##
wherein R.sup.1 and R.sup.2 are independently hydrogen,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkyl aryl, C.sub.1-C.sub.4
alkyl heteroaryl, C.sub.1-C.sub.4 alkyl heterocyclyl, or R.sup.1
and R.sup.2 together form a ring, either cycloalkyl or
heterocycloalkyl; wherein R.sup.3 is hydrogen, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 alkyl aryl, C.sub.1-C.sub.4 alkyl
heteroaryl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.1-C.sub.4 carbonyl, C.sub.1-C.sub.4 carbonylamino,
C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 sulfonyl, C.sub.1-C.sub.4
sulfonylamino, C.sub.1-C.sub.4 thioalkyl, or C.sub.1-C.sub.4
carboxyl, or R.sup.3 may from a ring with itself or R.sup.1 or
R.sup.2, and the stereocenters present, if any, being either `R` or
`S` in configuration independently; wherein X.sup.1, X.sup.2, and
X.sup.3 are, independently, hydrogen, hydroxyl, halogen,
C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4
alkynyl, amino, aminocarbonyl, nitro, cyano, C.sub.1-C.sub.4
carbonyl, C.sub.1-C.sub.4 carbonylamino, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.4 sulfonyl, C.sub.1-C.sub.4 sulfonylamino,
C.sub.1-C.sub.4 thioalkyl, or C.sub.r C.sub.4 carboxyl; and wherein
B is hydrogen, C.sub.1-C.sub.18 alkyl, C.sub.1-C.sub.18 alkyl aryl,
C.sub.1-C.sub.18 alkyl heteroaryl, C.sub.1-C.sub.18 carbonyl,
C.sub.1-C.sub.18 carbonylamino, C.sub.1-C.sub.18 sulfonyl,
C.sub.1-C.sub.18 sulfonylamino, or C.sub.1-C.sub.18 carboxyl, and
the stereocenters present, if any, being either `R` or `S` in
configuration independently.
[0073] Certain embodiments of Formula III include those wherein
R.sup.1, R.sup.2, and R.sup.3 are independently methyl or hydrogen,
and wherein X.sup.1, X.sup.2, and X.sup.3 are hydrogen. In another
preferred embodiment of Formula III, B is an aliphatic carbonyl
group. In another preferred embodiment of Formula III, B is a
benzoic carbonyl group.
[0074] In other embodiments of Formula III, B is an aliphatic
carbonyl group. In another preferred embodiment of Formula III, B
is a benzoic carbonyl group.
[0075] Certain embodiments of Formula I are compounds of Formula
IV:
##STR00004##
wherein R.sup.1 and R.sup.2 are independently hydrogen,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkyl aryl, C.sub.1-C.sub.4
alkyl heteroaryl, C.sub.1-C.sub.4 alkyl heterocyclyl, or R.sup.1
and R.sup.2 together form a ring, either cycloalkyl or
heterocyclyl; wherein R.sup.3 is a hydrogen, C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 alkyl aryl, C.sub.1-C.sub.4 alkyl heteroaryl,
C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 alkynyl, C.sub.1-C.sub.4
carbonyl, C.sub.1-C.sub.4 carbonylamino, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.4 sulfonyl, C.sub.1-C.sub.4 sulfonylamino,
C.sub.1-C.sub.4 thioalkyl, or C.sub.1-C.sub.4 carboxyl, and the
stereocenters present, if any, being either `R` or `S` in
configuration independently; wherein X.sup.1, X.sup.2, and X.sup.3
are, independently, hydrogen, hydroxyl, halogen, C.sub.1-C.sub.4
alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 alkynyl, amino,
aminocarbonyl, nitro, cyano, C.sub.1-C.sub.4 carbonyl,
C.sub.1-C.sub.4 carbonylamino, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.4 sulfonyl, C.sub.1-C.sub.4 sulfonylamino,
C.sub.1-C.sub.4 thioalkyl, or C.sub.1-C.sub.4 carboxyl; wherein B
is hydrogen, C.sub.1-C.sub.18 alkyl, C.sub.1-C.sub.18 alkyl aryl,
C.sub.1-C.sub.18 alkyl heteroaryl, C.sub.1-C.sub.18 carbonyl,
C.sub.1-C.sub.18 carbonylamino, C.sub.1-C.sub.18 sulfonyl,
C.sub.1-C.sub.18 sulfonylamino, or C.sub.1-C.sub.18 carboxyl, and
the stereocenters present, if any, being either `R` or `S` in
configuration independently; and wherein Z is a bond,
C.sub.1-C.sub.4 alkyl, heteroalkyl, or an O atom.
[0076] Certain embodiments of Formula IV are those wherein R.sup.1,
R.sup.2, and R.sup.3 are independently methyl or hydrogen, and
wherein X.sup.1, X.sup.2, and X.sup.3 are hydrogen.
[0077] In other embodiments of Formula IV, B is an aliphatic
carbonyl group. In other embodiments of Formula IV, B is a benzoic,
pyridyl, naphthyl, benzothiophene, or thiazole carbonyl group. In
other embodiments of Formula IV, B is C.sub.1-C.sub.2 alkyl aryl,
C.sub.2-carbonyl, C.sub.2-carbonyl amino, C.sub.2-carboxyl, or
C.sub.2-hydroxyl aryl.
[0078] Benzamide compounds according to the invention may include
those represented by Formula II:
##STR00005##
wherein R.sup.1, R.sup.2, and R.sup.3 are independently hydrogen,
C.sub.1-C.sub.4 alkyl, aryl, C.sub.1-C.sub.4 alkyl aryl,
C.sub.1-C.sub.4 alkyl heteroaryl, C.sub.1-C.sub.4 alkyl
heterocyclyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.1-C.sub.4 carbonyl, C.sub.1-C.sub.4 carbonylamino,
C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 sulfonyl, C.sub.1-C.sub.4
sulfonylamino, C.sub.1-C.sub.4 thioalkyl, C.sub.1-C.sub.4 carboxyl,
or form a ring with each other or with A; wherein A is
C.sub.1-C.sub.4 alkyl, or forms a ring structure with R.sup.1,
R.sup.2, or R.sup.3; wherein B is hydrogen, an aryl group, a
heteroaryl group, a cycloalkyl group, a heterocycloalkyl group,
C.sub.1-C.sub.22 alkyl, C.sub.1-C.sub.22 alkyl aryl,
C.sub.1-C.sub.22 alkyl heteroaryl, C.sub.2-C.sub.22 alkenyl,
C.sub.2-C.sub.22 alkynyl, C.sub.1-C.sub.22 carbonyl,
C.sub.1-C.sub.22 carbonylamino, C.sub.1-C.sub.22 alkoxy,
C.sub.1-C.sub.22 sulfonyl, C.sub.1-C.sub.22 sulfonylamino,
C.sub.1-C.sub.22 thioalkyl, or C.sub.1-C.sub.22 carboxyl, and the
stereocenters present, if any, being either `R` or `S` in
configuration independently; wherein X.sup.1, X.sup.2, and X.sup.3
are, independently, hydrogen, hydroxyl, halogen, C.sub.1-C.sub.4
alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 alkynyl, amino,
aminocarbonyl, nitro, cyano, C.sub.1-C.sub.4 carbonyl,
C.sub.1-C.sub.4 carbonylamino, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.4 sulfonyl, C.sub.1-C.sub.4 sulfonylamino,
C.sub.1-C.sub.4 thioalkyl, or C.sub.1-C.sub.4 carboxyl; wherein the
double circle
##STR00006##
indicates an aromatic or heteroaromatic ring; and wherein each Z is
independently a bond, C.sub.1-C.sub.4 alkyl, heteroalkyl, or an O
atom.
[0079] In Formula II, any and all of the stereocenters may be
independently either `R` or `S` in configuration.
[0080] In certain embodiments of Formula II, R.sup.1 is hydrogen or
C.sub.1-C.sub.4 alkyl; B is C.sub.1-C.sub.22 carbonyl; the
stereocenters present, if any, are either `R` or `S` in
configuration independently; X.sub.1, X.sub.2, and X.sub.3 are
independently hydrogen or halogen; the double circle
##STR00007##
indicates an aromatic or heteroaromatic ring; and Z is a bond,
C.sub.1-C.sub.4 alkyl, heteroalkyl, or an O atom.
[0081] Compounds according to the invention include those according
to Formula V:
##STR00008##
wherein Z is a bond or --CH.sub.2--; and B is an aryl group, a
heteroaryl group, a cycloalkyl group, a heterocycloalkyl group,
C.sub.1-C.sub.22 alkyl, C.sub.1-C.sub.22 alkyl aryl,
C.sub.1-C.sub.22 alkyl heteroaryl, C.sub.2-C.sub.22 alkenyl,
C.sub.2-C.sub.22 alkynyl, C.sub.1-C.sub.22 carbonyl,
C.sub.1-C.sub.22 carbonylamino, C.sub.1-C.sub.22 alkoxy,
C.sub.1-C.sub.22 sulfonyl, C.sub.1-C.sub.22 sulfonylamino,
C.sub.1-C.sub.22 thioalkyl, or C.sub.1-C.sub.22 carboxyl.
[0082] Compounds according to the invention include those according
to Formula (VI):
##STR00009##
wherein R.sup.4 is C.sub.1-C.sub.4 alkyl and n is 0-3.
[0083] In some embodiments, R.sup.4 is methyl and n is 2.
[0084] Compounds according to the invention may include salts and
solvates of the compounds according to Formulas I, II, III, IV, V,
or VI. When racemates exists, each enantiomer or diastereomer may
be separately used, or they may be combined in any proportion.
Where tautomers exist all possible tautomers are specifically
contemplated.
[0085] Compounds according to the invention include compounds
E482-E496 shown below:
##STR00010## ##STR00011## ##STR00012## ##STR00013##
##STR00014##
Synthesis of Compounds
[0086] The amino isoquinoline amide or substituted benzamide
compounds may be synthesized according to the general schemes shown
in FIGS. 1-5.
[0087] According to the synthesis scheme in FIG. 1, ester (1) may
be protected with the TIPS group and alkylated with
bromomethylphthalimide to give compound (3). The ester may be then
hydrolyzed with LiOH*H.sub.2O to give diacid (4) and coupled with
6-aminoisoquinoline using EDC as the coupling agent. The amine (6)
may be accomplished using hydrazine which may be then protected
with Boc.sub.2O to give (7). Deprotection of the hydroxyl group may
be carried out with TBAF, and coupling with the appropriate acid
may be achieved with EDC or using the acid chloride. Deprotection
of the amine may be accomplished with HCl to give the final amino
isoquinoline amides.
[0088] As in FIG. 1, the synthesis scheme in FIG. 2 may begin by
protecting 2-(4-(hydroxymethyl)phenyl) acetic acid as the methyl
ester and the TIPS alcohol to give E457. This methyl ester may be
then alkylated with bromomethylphthalimide to give compound E459.
The ester may be hydrolyzed with LiOH*H.sub.2O to give diacid E460
and coupled with 6-aminoisoquinoline using EDC as the coupling
agent giving compound E461. Formation of the amine E462 may be
accomplished using hydrazine which may be then protected with
Boc.sub.2O to give E463. Deprotection of the hydroxyl group may be
carried out with TBAF, and coupling with the appropriate acid may
be achieved with EDC or using the acid chloride. Deprotection of
the amine may be accomplished with HCl to give the final amino
isoquinoline amides.
[0089] Benzamides may be synthesized using the procedures outlined
in FIG. 2, but by substituting the para-amino benzamide of choice
for the amino isoquinoline, as shown in the synthesis scheme in
FIG. 3.
[0090] The para-aminobenzamide precursors of the synthesis scheme
in FIG. 3 may be commercially-available, or may be synthesized by
the general synthesis schemes of FIGS. 4-5.
[0091] According to FIG. 4, the appropriate acid may be converted
to its acid chloride with oxalyl chloride then reacted with ammonia
gas or another amine to give the amide. The nitro group may be
reduced to the aniline with hydrogen or another reducing agent. The
aniline may be then coupled with an appropriate acid using standard
coupling procedures such as EDC and DMAP in pyridine as shown in
FIG. 3.
[0092] An alternative synthetic route is outlined in the synthesis
scheme of FIG. 5. According to FIG. 5, the aniline may be coupled
with an appropriate acid using standard coupling procedures such as
EDC and DMAP in pyridine. The ester may be then converted to the
corresponding primary amide using formamide and NaOMe in DMF or to
a substituted amide by heating with the appropriate amine in a
solvent such as MeOH.
[0093] The abbreviations used in the synthetic schemes shown in the
figures have the following meanings: Boc.sub.2O is
di-tert-butyl-dicarbonate, DMAP is dimethyl aminopyridine, DMSO is
Dimethyl Sulfoxide, HATU is
2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate, LDA is lithium diisopropyl amide, DMF is
dimethylformamide, THF is tetrahydrofuran, and EDC is
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride.
Compositions
[0094] The compounds of Formulas I-VI may be provided or formulated
in a composition. The compounds may be administered in a
pharmaceutically acceptable formulation, such as in or with a
pharmaceutically acceptable carrier.
[0095] Compounds according to the invention may be administered in
conjunction with one or more additional therapeutic agents.
Suitable additional therapeutic agents may include, but are not
limited to, beta blockers, alpha-agonists, carbonic anhydrase
inhibitors, prostaglandin-like compounds, miotic or cholinergic
agents, or epinephrine compounds.
[0096] Beta blockers reduce the production of aqueous humor.
Examples include levobunolol (BETAGAN.RTM.), timolol (BETIMOL.RTM.,
TIMOPTIC.RTM.), betaxolol (BETOPTIC.RTM.) and metipranolol
(OPTIPRANOLOL.RTM.).
[0097] Alpha-agonists reduce the production of aqueous humor and
increase drainage. Examples include apraclonidine (IOPIDINE.RTM.)
and brimonidine (ALPHAGAN.RTM.).
[0098] Carbonic anhydrase inhibitors reduce the production of
aqueous humor. Examples include dorzolamide (TRUSOPT.RTM.) and
brinzolamide (AZOPT.RTM.).
[0099] Prostaglandins and prostaglandin-like compounds increase the
outflow of aqueous humor. Examples include latanoprost
(XALATAN.RTM.), bimatoprost (LUMIGAN.RTM.), and travoprost
(TRAVATANT.TM.).
[0100] Miotic or cholinergic agents increase the outflow of aqueous
humor. Examples include pilocarpine (ISOPTO CARPINE.RTM.,
PILOPINE.RTM.) and carbachol (ISOPTO CARBACHOL.RTM.).
[0101] Epinephrine compounds, such as dipivefrin (PROPINE.RTM.),
also increase the outflow of aqueous humor.
[0102] The additional therapeutic agent or agents may be
administered simultaneously or sequentially with the compounds of
the present invention. Sequential administration includes
administration before or after the compounds of the present
invention. In some embodiments, the additional therapeutic agent or
agents may be administered in the same composition as the compounds
of the present invention. In other embodiments, there may be an
interval of time between administration of the additional
therapeutic agent and the compounds of the present invention.
[0103] In some embodiments, the administration of an additional
therapeutic agent with a compound of the present invention may
enable lower doses of the other therapeutic agents and/or
administration at less frequent intervals.
[0104] Pharmaceutical compositions for use in accordance with the
present invention may be formulated in a conventional manner using
one or more physiologically acceptable carriers or excipients.
Thus, the compounds and their physiologically acceptable salts and
solvates may be formulated for administration by, for example,
solid dosing, eyedrop, in a topical oil-based formulation,
injection, inhalation (either through the mouth or the nose),
implants, or oral, buccal, parenteral, or rectal administration.
Techniques and formulations may generally be found in
"Reminington's Pharmaceutical Sciences", (Meade Publishing Co.,
Easton, Pa.). Therapeutic compositions must typically be sterile
and stable under the conditions of manufacture and storage.
[0105] Compositions of the present invention may comprise a safe
and effective amount of the subject compounds, and a
pharmaceutically-acceptable carrier. As used herein, "safe and
effective amount" means an amount of a compound sufficient to
significantly induce a positive modification in the condition to be
treated, but low enough to avoid serious side effects (at a
reasonable benefit/risk ratio), within the scope of sound medical
judgment. A safe and effective amount of a compound will vary with
the particular condition being treated, the age and physical
condition of the patient being treated, the severity of the
condition, the duration of treatment, the nature of concurrent
therapy, the particular pharmaceutically-acceptable carrier
utilized, and like factors within the knowledge and expertise of
the attending physician.
[0106] The route by which the compounds of the present invention
(component A) will be administered and the form of the composition
will dictate the type of carrier (component B) to be used. The
composition may be in a variety of forms, suitable, for example,
for systemic administration (e.g., oral, rectal, nasal, sublingual,
buccal, implants, or parenteral) or topical administration (e.g.,
dermal, pulmonary, nasal, aural, ocular, liposome delivery systems,
or iontophoresis).
[0107] Carriers for systemic administration typically comprise at
least one of a) diluents, b) lubricants, c) binders, d)
disintegrants, e) colorants, f) flavors, g) sweeteners, h)
antioxidants, j) preservatives, k) glidants, m) solvents, n)
suspending agents, o) wetting agents, p) surfactants, combinations
thereof, and others. All carriers are optional in the systemic
compositions.
[0108] Ingredient a) is a diluent. Suitable diluents for solid
dosage forms include sugars such as glucose, lactose, dextrose, and
sucrose; diols such as propylene glycol; calcium carbonate; sodium
carbonate; sugar alcohols, such as glycerin; mannitol; and
sorbitol. The amount of ingredient a) in the systemic or topical
composition is typically about 50 to about 90%.
[0109] Ingredient b) is a lubricant. Suitable lubricants for solid
dosage forms are exemplified by solid lubricants including silica,
talc, stearic acid and its magnesium salts and calcium salts,
calcium sulfate; and liquid lubricants such as polyethylene glycol
and vegetable oils such as peanut oil, cottonseed oil, sesame oil,
olive oil, corn oil and oil of theobroma. The amount of ingredient
b) in the systemic or topical composition is typically about 5 to
about 10%.
[0110] Ingredient c) is a binder. Suitable binders for solid dosage
forms include polyvinyl pyrrolidone; magnesium aluminum silicate;
starches such as corn starch and potato starch; gelatin;
tragacanth; and cellulose and its derivatives, such as sodium
carboxymethylcellulose, ethyl cellulose, methylcellulose,
microcrystalline cellulose, and sodium carboxymethylcellulose. The
amount of ingredient c) in the systemic composition is typically
about 5 to about 50%, and in ocular solid dosing forms up to
99%.sub..
[0111] Ingredient d) is a disintegrant. Suitable disintegrants for
solid dosage forms include agar, alginic acid and the sodium salt
thereof, effervescent mixtures, croscarmelose, crospovidone, sodium
carboxymethyl starch, sodium starch glycolate, clays, and ion
exchange resins. The amount of ingredient d) in the systemic or
topical composition is typically about 0.1 to about 10%.
[0112] Ingredient e) for solid dosage forms is a colorant such as
an FD&C dye. When used, the amount of ingredient e) in the
systemic or topical composition is typically about 0.005 to about
0.1%.
[0113] Ingredient f) for solid dosage forms is a flavor such as
menthol, peppermint, and fruit flavors. The amount of ingredient
f), when used, in the systemic or topical composition is typically
about 0.1 to about 1.0%.
[0114] Ingredient g) for solid dosage forms is a sweetener such as
aspartame and saccharin. The amount of ingredient g) in the
systemic or topical composition is typically about 0.001 to about
1%.
[0115] Ingredient h) is an antioxidant such as butylated
hydroxyanisole ("BHA"), butylated hydroxytoluene ("BHT"), and
vitamin E. The amount of ingredient h) in the systemic or topical
composition is typically about 0.1 to about 5%.
[0116] Ingredient j) is a preservative such as benzalkonium
chloride, methyl paraben and sodium benzoate. The amount of
ingredient j) in the systemic or topical composition is typically
about 0.01 to about 5%.
[0117] Ingredient k) for solid dosage forms is a glidant such as
silicon dioxide. The amount of ingredient k) in the systemic or
topical composition is typically about 1 to about 5%.
[0118] Ingredient m) is a solvent, such as water, isotonic saline,
ethyl oleate, glycerine, hydroxylated castor oils, alcohols such as
ethanol, and phosphate buffer solutions. The amount of ingredient
m) in the systemic or topical composition is typically from about 0
to about 100%.
[0119] Ingredient n) is a suspending agent. Suitable suspending
agents include AVICEL.RTM. RC-591 (from FMC Corporation of
Philadelphia, Pa.) and sodium alginate. The amount of ingredient n)
in the systemic or topical composition is typically about 1 to
about 8%.
[0120] Ingredient o) is a surfactant such as lecithin, Polysorbate
80, and sodium lauryl sulfate, and the TWEENS.RTM. from Atlas
Powder Company of Wilmington, Del. Suitable surfactants include
those disclosed in the C.T.F.A. Cosmetic Ingredient Handbook, 1992,
pp. 587-592; Remington's Pharmaceutical Sciences, 15th Ed. 1975,
pp. 335-337; and McCutcheon's Volume 1, Emulsifiers &
Detergents, 1994, North American Edition, pp. 236-239. The amount
of ingredient o) in the systemic or topical composition is
typically about 0.1% to about 5%.
[0121] Although the amounts of components A and B in the systemic
compositions may vary depending on the type of systemic composition
prepared, the specific derivative selected for component A and the
ingredients of component B, in general, system compositions
comprise 0.01% to 50% of component A and 50% to 99.99% of component
B.
[0122] Compositions for parenteral administration typically
comprise A) 0.1% to 10% of the compounds of the present invention
and B) 90% to 99.9% of a carrier comprising a) a diluent and m) a
solvent. In one embodiment, component a) comprises propylene glycol
and m) comprises ethanol or ethyl oleate.
[0123] Compositions for oral administration can have various dosage
forms. For example, solid forms include tablets, capsules,
granules, and bulk powders. These oral dosage forms comprise a safe
and effective amount, usually at least about 5%, and more
particularly from about 25% to about 50% of component A). The oral
dosage compositions further comprise about 50% to about 95% of
component B), and more particularly, from about 50% to about
75%.
[0124] Tablets can be compressed, tablet triturates,
enteric-coated, sugar-coated, film-coated, or multiple-compressed.
Tablets typically comprise component A, and component B a carrier
comprising ingredients selected from the group consisting of a)
diluents, b) lubricants, c) binders, d) disintegrants, e)
colorants, f) flavors, g) sweeteners, k) glidants, and combinations
thereof. Specific diluents include calcium carbonate, sodium
carbonate, mannitol, lactose and cellulose. Specific binders
include starch, gelatin, and sucrose. Specific disintegrants
include alginic acid and croscarmelose. Specific lubricants include
magnesium stearate, stearic acid, and talc. Specific colorants are
the FD&C dyes, which can be added for appearance. Chewable
tablets preferably contain g) sweeteners such as aspartame and
saccharin, or f) flavors such as menthol, peppermint, fruit
flavors, or a combination thereof.
[0125] Capsules (including implants, time release and sustained
release formulations) typically comprise component A, and a carrier
comprising one or more a) diluents disclosed above in a capsule
comprising gelatin. Granules typically comprise component A, and
preferably further comprise k) glidants such as silicon dioxide to
improve flow characteristics. Implants can be of the biodegradable
or the non-biodegradable type. Implants may be prepared using any
known biocompatible formulation.
[0126] The selection of ingredients in the carrier for oral
compositions depends on secondary considerations like taste, cost,
and shelf stability, which are not critical for the purposes of
this invention. One skilled in the art would know how to select
appropriate ingredients without undue experimentation.
[0127] The solid compositions may also be coated by conventional
methods, typically with pH or time-dependent coatings, such that
component A is released in the gastrointestinal tract in the
vicinity of the desired application, or at various points and times
to extend the desired action. The coatings typically comprise one
or more components selected from the group consisting of cellulose
acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl
methyl cellulose phthalate, ethyl cellulose, EUDRAGIT.RTM. coatings
(available from Rohm & Haas G.M.B.H. of Darmstadt, Germany),
waxes and shellac.
[0128] Compositions for oral administration can also have liquid
forms. For example, suitable liquid forms include aqueous
solutions, emulsions, suspensions, solutions reconstituted from
non-effervescent granules, suspensions reconstituted from
non-effervescent granules, effervescent preparations reconstituted
from effervescent granules, elixirs, tinctures, syrups, and the
like. Liquid orally administered compositions typically comprise
component A and component B, namely, a carrier comprising
ingredients selected from the group consisting of a) diluents, e)
colorants, f) flavors, g) sweeteners, j) preservatives, m)
solvents, n) suspending agents, and o) surfactants. Peroral liquid
compositions preferably comprise one or more ingredients selected
from the group consisting of e) colorants, f) flavors, and g)
sweeteners.
[0129] Other compositions useful for attaining systemic delivery of
the subject compounds include sublingual, buccal and nasal dosage
forms. Such compositions typically comprise one or more of soluble
filler substances such as a) diluents including sucrose, sorbitol
and mannitol; and c) binders such as acacia, microcrystalline
cellulose, carboxymethyl cellulose, and hydroxypropyl
methylcellulose. Such compositions may further comprise b)
lubricants, e) colorants, f) flavors, g) sweeteners, h)
antioxidants, and k) glidants.
[0130] In one embodiment of the invention, the compounds of the
present invention are topically administered. Topical compositions
that can be applied locally to the eye may be in any form known in
the art, non-limiting examples of which include solids, liquid
drops, gelable drops, sprays, ointments, or a sustained or
non-sustained release unit placed in the conjunctival cul-du-sac of
the eye or another appropriate location.
[0131] Topical compositions that can be applied locally to the skin
may be in any form including solids, solutions, oils, creams,
ointments, gels, lotions, shampoos, leave-on and rinse-out hair
conditioners, milks, cleansers, moisturizers, sprays, skin patches,
and the like. Topical compositions comprise: component A, the
compounds described above, and component B, a carrier. The carrier
of the topical composition preferably aids penetration of the
compounds into the skin. Component B may further comprise one or
more optional components.
[0132] The amount of the carrier employed in conjunction with
component A is sufficient to provide a practical quantity of
composition for administration per unit dose of the medicament.
Techniques and compositions for making dosage forms useful in the
methods of this invention are described in the following
references: Modern Pharmaceutics, Chapters 9 and 10, Banker &
Rhodes, eds. (1979); Lieberman et al., Pharmaceutical Dosage Forms:
Tablets (1981); and Ansel, Introduction to Pharmaceutical Dosage
Forms, 2.sup.nd Ed., (1976).
[0133] Component B may comprise a single ingredient or a
combination of two or more ingredients. In the topical
compositions, component B comprises a topical carrier. Suitable
topical carriers comprise one or more ingredients selected from the
group consisting of phosphate buffered saline, isotonic water,
deionized water, monofunctional alcohols, symmetrical alcohols,
aloe vera gel, allantoin, glycerin, vitamin A and E oils, mineral
oil, propylene glycol, PPG-2 myristyl propionate, dimethyl
isosorbide, castor oil, combinations thereof, and the like. More
particularly, carriers for skin applications include propylene
glycol, dimethyl isosorbide, and water, and even more particularly,
phosphate buffered saline, isotonic water, deionized water,
monofunctional alcohols, and symmetrical alcohols.
[0134] The carrier of the topical composition may further comprise
one or more ingredients selected from the group consisting of q)
emollients, r) propellants, s) solvents, t) humectants, u)
thickeners, v) powders, w) fragrances, x) pigments, and y)
preservatives.
[0135] Ingredient q) is an emollient. The amount of ingredient q)
in a skin-based topical composition is typically about 5% to about
95%. Suitable emollients include stearyl alcohol, glyceryl
monoricinoleate, glyceryl monostearate, propane-1,2-diol,
butane-1,3-diol, mink oil, cetyl alcohol, isopropyl isostearate,
stearic acid, isobutyl palmitate, isocetyl stearate, oleyl alcohol,
isopropyl laurate, hexyl laurate, decyl oleate, octadecan-2-ol,
isocetyl alcohol, cetyl palmitate, di-n-butyl sebacate, isopropyl
myristate, isopropyl palmitate, isopropyl stearate, butyl stearate,
polyethylene glycol, triethylene glycol, lanolin, sesame oil,
coconut oil, arachis oil, castor oil, acetylated lanolin alcohols,
petroleum, mineral oil, butyl myristate, isostearic acid, palmitic
acid, isopropyl linoleate, lauryl lactate, myristyl lactate, decyl
oleate, myristyl myristate, and combinations thereof. Specific
emollients for skin include stearyl alcohol and
polydimethylsiloxane.
[0136] Ingredient r) is a propellant. The amount of ingredient r)
in the topical composition is typically about 0% to about 95%.
Suitable propellants include propane, butane, isobutane, dimethyl
ether, carbon dioxide, nitrous oxide, and combinations thereof.
[0137] Ingredient s) is a solvent. The amount of ingredient s) in
the topical composition is typically about 0% to about 95%.
Suitable solvents include water, ethyl alcohol, methylene chloride,
isopropanol, castor oil, ethylene glycol monoethyl ether,
diethylene glycol monobutyl ether, diethylene glycol monoethyl
ether, dimethylsulfoxide, dimethyl formamide, tetrahydrofuran, and
combinations thereof. Specific solvents include ethyl alcohol and
homotopic alcohols.
[0138] Ingredient t) is a humectant. The amount of ingredient t) in
the topical composition is typically 0% to 95%. Suitable humectants
include glycerin, sorbitol, sodium 2-pyrrolidone-5-carboxylate,
soluble collagen, dibutyl phthalate, gelatin, and combinations
thereof. Specific humectants include glycerin.
[0139] Ingredient u) is a thickener. The amount of ingredient u) in
the topical composition is typically about 0% to about 95%.
[0140] Ingredient v) is a powder. The amount of ingredient v) in
the topical composition is typically 0% to 95%. Suitable powders
include beta-cyclodextrins, hydroxypropyl cyclodextrins, chalk,
talc, fullers earth, kaolin, starch, gums, colloidal silicon
dioxide, sodium polyacrylate, tetra alkyl ammonium smectites,
trialkyl aryl ammonium smectites, chemically-modified magnesium
aluminum silicate, organically-modified Montmorillonite clay,
hydrated aluminum silicate, fumed silica, carboxyvinyl polymer,
sodium carboxymethyl cellulose, ethylene glycol monostearate, and
combinations thereof. For ocular applications, specific powders
include beta-cyclodextrin, hydroxypropyl cyclodextrin, and sodium
polyacrylate. For gel dosing ocular formulations, sodium
polyacrylate may be used.
[0141] Ingredient w) is a fragrance. The amount of ingredient w) in
the topical composition is typically about 0% to about 0.5%,
particularly, about 0.001% to about 0.1%. For ocular applications a
fragrance is not typically used.
[0142] Ingredient x) is a pigment. Suitable pigments for skin
applications include inorganic pigments, organic lake pigments,
pearlescent pigments, and mixtures thereof. Inorganic pigments
useful in this invention include those selected from the group
consisting of rutile or anatase titanium dioxide, coded in the
Color Index under the reference CI 77,891; black, yellow, red and
brown iron oxides, coded under references CI 77,499, 77,492 and,
77,491; manganese violet (CI 77,742); ultramarine blue (CI 77,007);
chromium oxide (CI 77,288); chromium hydrate (CI 77,289); and
ferric blue (CI 77,510) and mixtures thereof.
[0143] The organic pigments and lakes useful in this invention
include those selected from the group consisting of D&C Red No.
19 (CI 45,170), D&C Red No. 9 (CI 15,585), D&C Red No. 21
(CI 45,380), D&C Orange No. 4 (CI 15,510), D&C Orange No.
(CI 45,370), D&C Red No. 27 (CI 45,410), D&C Red No. 13 (CI
15,630), D&C Red No. 7 (CI 15,850), D&C Red No. 6 (CI
15,850), D&C Yellow No. 5 (CI 19,140), D&C Red No. 36 (CI
12,085), D&C Orange No. 10 (CI 45,425), D&C Yellow No. 6
(CI 15,985), D&C Red No. 30 (CI 73,360), D&C Red No. 3 (CI
45,430), the dye or lakes based on Cochineal Carmine (CI 75,570)
and mixtures thereof.
[0144] The pearlescent pigments useful in this invention include
those selected from the group consisting of the white pearlescent
pigments such as mica coated with titanium oxide, bismuth
oxychloride, colored pearlescent pigments such as titanium mica
with iron oxides, titanium mica with ferric blue, chromium oxide
and the like, titanium mica with an organic pigment of the
above-mentioned type as well as those based on bismuth oxychloride
and mixtures thereof. The amount of pigment in the topical
composition is typically about 0% to about 10%. For ocular
applications a pigment is generally not used.
[0145] In other certain embodiments of the invention, topical
pharmaceutical compositions for ocular administration are prepared
typically comprising component A and B (a carrier), such as
purified water, and one or more ingredients selected from the group
consisting of y) sugars or sugar alcohols such as dextrans,
particularly mannitol and dextran 70, z) cellulose or a derivative
thereof, aa) a salt, bb) disodium EDTA (Edetate disodium), and cc)
a pH adjusting additive.
[0146] Examples of z) cellulose derivatives suitable for use in the
topical pharmaceutical composition for ocular administration
include sodium carboxymethylcellulose, ethylcellulose,
methylcellulose, and hydroxypropyl-methylcellulose, particularly,
hydroxypropyl-methylcellulose.
[0147] Examples of aa) salts suitable for use in the topical
pharmaceutical composition for ocular administration include mono-,
di- and trisodium phosphate, Tris(hydroxymethyl) aminomethane
hydrochloride, sodium chloride, potassium chloride, and
combinations thereof.
[0148] Examples of cc) pH adjusting additives include HCl or NaOH
in amounts sufficient to adjust the pH of the topical
pharmaceutical composition for ocular administration to
5.0-7.5.
[0149] Component A may be included in kits comprising component A,
a systemic or topical composition described above, or both; and
information, instructions, or both that use of the kit will provide
treatment for cosmetic and medical conditions in mammals
(particularly humans). The information and instructions may be in
the form of words, pictures, or both, and the like. In addition or
in the alternative, the kit may comprise the medicament, a
composition, or both; and information, instructions, or both,
regarding methods of application of medicament, or of composition,
preferably with the benefit of treating or preventing cosmetic and
medical conditions in mammals (e.g., humans).
Methods of Use
[0150] Compounds according to Formulas I-VI and compositions
including them may have kinase inhibitory activity and thus may be
useful in influencing or inhibiting the action of kinases, and in
the treatment and/or prevention of diseases or conditions
influenced by kinases. At a minimum, they may have NET activity and
this may be useful for influencing the action of NET. In some
embodiments, methods are provided that may comprise administering a
composition comprising a specific ligand that interacts strongly
with a kinase, specifically and at minimum a rho kinase, and that
also interacts with monoamine transporter proteins (MATs),
specifically and at minimum with NET proteins.
[0151] According to certain embodiments, provided are methods of
influencing and/or inhibiting the action of a kinase in a cell or
medium. In certain embodiments, provided are methods of influencing
and/or inhibiting NET in a cell or medium. A cell may be in vitro,
in a body in vivo, or in a living body in vivo. A medium may
include an assay medium. The methods may comprise applying to a
medium or contacting a cell with an effective amount of a compound
according to Formula I, II, III, IV, V, or VI. The methods may
comprise applying to a medium or contacting a cell with composition
comprising an effective amount of a compound according to Formula
I, II, III, IV, V, or VI, and a pharmaceutically acceptable
carrier. In some embodiments, the kinase inhibited may be a rho
kinase.
[0152] According to certain embodiments, provided are methods of
treating a disease, disorder, or condition. The methods may
comprise administering to a subject a compound according to Formula
I, II, III, IV, V, or VI. The methods may comprise administering to
a subject a composition comprising a compound according to Formula
I, II, III, IV, V, or VI, and a pharmaceutically acceptable
carrier. Diseases, disorders, or conditions may include any of the
diseases or conditions associated with kinase activity or diseases
or conditions affected by kinases. For example, the disease may be
selected from the group consisting of eye disease including
glaucoma and retinal diseases such as Wet AMD Dry AMD
(inflammation) and DME, bone disorder including osteoporosis,
vascular disease including cerebral vasospasm, coronary vasospasm,
hypertension, pulmonary hypertension, sudden death syndrome,
angina, myocardial infarction, restenosis, stroke, hypertensive
vascular disease, heart failure, cardiac allograft vasculopathy,
vein graft disease, pulmonary disease including chronic obstructive
pulmonary disease (COPD) and asthma, neurological disorder
including spinal cord injury, Alzheimer's disease, multiple
sclerosis, depression, attention deficit-hyperactivity disorder and
neuropathic pain, neovascular disorders and cancer, obesity, and
erectile dysfunction. The compounds of the present invention may
also be useful in decreasing intraocular pressure. Thus, these
compounds may be useful in the treatment of glaucoma. The preferred
route of administration for treating glaucoma is topically.
[0153] In further aspects, the invention provides methods of
modulating kinase activity, the methods comprising contacting a
cell with a compound according to Formula I, II, III, IV, V, or VI
as described above, in an amount effective to modulate kinase
activity.
[0154] In further aspects, the invention provides methods of
reducing intraocular pressure, the methods comprising contacting a
cell with a compound according to Formula I, II, III, IV, V, or VI
as described above, in an amount effective to reduce intraocular
pressure.
[0155] The dosage range of the compound for systemic administration
is from about 0.001 to about 100 mg/kg body weight, preferably from
about 0.01 to about 10 mg/kg per body weight, most preferably form
about 0.05 to about 5 mg/kg body weight per day. The transdermal
dosages will be designed to attain similar serum or plasma levels,
based upon techniques known to those skilled in the art of
pharmacokinetics and transdermal formulations. Plasma levels for
systemic administration are expected to be in the range of 0.1 to
1000 ng/mL, more preferably from 0.5 to 500 ng/mL and most
preferably from 1 to 100 ng/mL. While these dosages are based upon
a daily administration rate, weekly or monthly accumulated dosages
may also be used to calculate the clinical requirements.
[0156] Dosages may be varied based on the patient being treated,
the condition being treated, the severity of the condition being
treated, the route of administration, etc. to achieve the desired
effect.
[0157] The invention will be further explained by the following
illustrative examples that are to be considered to be
non-limiting.
EXAMPLES
Reference Example 1
[0158] All temperatures were in degrees Centigrade. Reagents and
starting materials were purchased from commercial sources or
prepared following published literature procedures.
[0159] Unless otherwise noted, HPLC purification, when appropriate,
was performed by redissolving the compound in a small volume of
DMSO and filtering through a 0.45 micron (nylon disc) syringe
filter. The solution was then purified using, for example, a 50 mm
Varian Dynamax HPLC 21.4 mm Microsorb Guard-8 C.sub.8 column. A
typical initial eluting mixture of 40-80% MeOH:H.sub.2O was
selected as appropriate for the target compound. This initial
gradient was maintained for 0.5 min then increased to 100% MeOH:0%
H.sub.2O over 5 min. 100% MeOH was maintained for 2 more min before
re-equilibration back to the initial starting gradient. A typical
total run time was 8 min. The resulting fractions were analyzed,
combined as appropriate, and then evaporated to provide purified
material.
[0160] Proton magnetic resonance (.sup.1H NMR) spectra were
recorded on either a Varian INOVA 600 MHz (.sup.1H) NMR
spectrometer, Varian INOVA 500 MHz (.sup.1H) NMR spectrometer,
Varian Mercury 300 MHz (.sup.1H) NMR spectrometer, or a Varian
Mercury 200 MHz (.sup.1H) NMR spectrometer. All spectra were
determined in the solvents indicated. Although chemical shifts are
reported in ppm downfield of tetramethylsilane, they are referenced
to the residual proton peak of the respective solvent peak for
.sup.1H NMR. Interproton coupling constants are reported in Hertz
(Hz).
[0161] Analytical LCMS spectra were obtained using a Waters ZQ MS
ESI instrument with an Alliance 2695 HPLC and a 2487 dual
wavelength UV detector. Spectra were analyzed at 254 and 230 nm.
Samples were passed through a Waters Symmetry C18 4.6.times.75 mm
3.5 .mu.column with or without a guard column (3.9.times.20 mm
5.mu.). Gradients were run with mobile phase A: 0.1% formic acid in
H.sub.2O and mobile phase B: ACN with a flow rate of 0.8 mL/min.
Two gradients will illustrate:
TABLE-US-00001 Gradient A Gradient B Time A % B % Time A % B % 0.00
80.0 20.0 0.00 95.0 20.0 1.00 80.0 20.0 1.00 9.0 25.0 6.00 25.0
75.0 6.00 40.0 75.0 7.00 5.0 95.0 7.00 5.0 95.0 8.00 5.0 95.0 8.00
5.0 95.0 9.00 80.0 20.0 9.00 95.0 20.0 12.00 80.0 20.0 12.00 95.0
20.0
[0162] The settings for the MS probe were a cone voltage at 38 mV
and a desolvation temperature at 250.degree. C. Any variations in
these methods are noted below.
[0163] The following preparations illustrate procedures for the
preparation of intermediates and methods for the preparation of an
amino isoquinoline amide derivatives or substituted benzamide
derivatives.
Examples 1-12
[0164] Compounds E1-E12 may be synthesized according to the scheme
shown in FIG. 1 and FIG. 6. For example, methyl
2-(4-(triisopropylsilyloxy)phenyl)acetate (E2) was synthesized from
E1 according to the below:
##STR00015##
To methyl 2-(4-hydroxyphenyl)acetate (E1) in CH.sub.2Cl.sub.2 at
0.degree. C. was added 2,6-lutidine and TIPS-OTf. The ice bath was
removed and the solution was allowed to warm to room temperature
and stirred. After 4 h the solution was poured into
NH.sub.4Cl.sub.(sat) and CH.sub.2Cl.sub.2 and the organic layer was
further extracted with NH.sub.4Cl.sub.(sat). The organics were
dried (Na.sub.2SO.sub.4) filtered and evaporated. Column
chromatography (0-15% EtOAc/Hexanes) gave pure
methyl-2-(4-(triisopropylsilyloxy)phenyl)acetate (E2).
[0165] Methyl
3-(1,3-dioxoisoindolin-2-yl)-2-(4-(triisopropylsilyloxy)phenyl)
propanoate (E3) was prepared from E2 according to the below:
##STR00016##
To a solution of LiHMDS in THF cooled to -78.degree. C. was added a
cooled solution (approx -78.degree. C.) of
methyl-2-(4-(triisopropylsilyloxy) phenyl)acetate (E2) in THF via
syringe. The solution was stirred at -78.degree. C. for 30 min.
Bromo-methyl phthalimide was added directly to the anion, and the
solution was immediately removed from the -78.degree. C. bath and
placed in an ice bath and stirred for 2 h. The reaction was then
poured into NH.sub.4Cl.sub.(sat) and extracted with EtOAc. The
organics were dried (Na.sub.2SO.sub.4), filtered, and evaporated.
Column chromatography 0-20% EtOAc/Hexanes gave pure methyl
3-(1,3-dioxoisoindolin-2-yl)-2-(4-(triisopropylsilyloxy)phenyl)propanoate
(E3).
[0166]
2-(2-Carboxy-2-(4-(triisopropylsilyloxy)phenyl)ethylcarbamoyl)
benzoic acid (E4) was prepared from E3 according to the below:
##STR00017##
To methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(4-(triisopropylsilyloxy)
phenyl) propanoate (E3) in THF/H.sub.2O was added LiOH.H.sub.2O,
and the solution was stirred for 1.5 h or until conversion to
product was visible by LC-MS. The solution was then poured into
EtOAc/NH.sub.4Cl(sat)/1 N HCl (3:1), and the aqueous layer was
further extracted with EtOAc. The organics were dried
(Na.sub.2SO.sub.4), filtered, evaporated, and dried to give crude
2-(2-carboxy-2-(4-(triisopropylsilyloxy)phenyl)ethylcarbamoyl)benzoic
acid (E4).
[0167]
3-(1,3-Dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-(triisopropy-
lsilyloxy phenyl) propanamide (E5) was prepared from E4 according
to the below:
##STR00018##
To 2-(2-carboxy-2-(4-(triisopropylsilyloxy)phenyl)ethylcarbamoyl)
benzoic acid (E4) in pyridine was added EDC, DMAP, and
6-aminoisoquinoline, and the solution was flushed with N.sub.2,
capped, and stirred overnight. The mixture was poured into
EtOAc/NaHCO.sub.3(sat) and the aqueous layer was further extracted
with EtOAc. The organics were dried (Na.sub.2SO.sub.4), filtered,
and evaporated. Column chromatography 5% MeOH/CH.sub.2Cl.sub.2 gave
pure
3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-(triisop-
ropylsilyloxy phenyl) propanamide (E5).
[0168]
3-amino-N-(isoquinolin-6-yl)-2-(4-(triisopropylsilyloxy)phenyl)prop-
anamide (E6) was prepared from E5 according to the below:
##STR00019##
To
3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-(triisopropylsi-
lyloxy phenyl) propanamide (E5) in EtOH was added
NH.sub.2--NH.sub.2, and the solution was refluxed for 1.2 hrs-2
hrs. The solids were filtered and the solvents were evaporated.
Column chromatography 5% 2N NH.sub.3-MeOH/CH.sub.2Cl.sub.2 gave
pure
3-amino-N-(isoquinolin-6-yl)-2-(4-(triisopropylsilyloxy)phenyl)propanamid-
e (E6).
[0169] Tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(triisopropylsilyloxy) phenyl)
propyl carbamate (E7) was prepared from E6 according to the
below:
##STR00020##
To 3-amino-N-(isoquinolin-6-yl)-2-(4-(triisopropylsilyloxy)
phenyl)propanamide (E6) in CH.sub.2Cl.sub.2 (7.3 mL) at 0.degree.
C. was added a solution of Boc.sub.2O in CH.sub.2Cl.sub.2 also
cooled to 0.degree. C. before addition. The solution stirred for 30
min at 0.degree. C. and additional Boc.sub.2O was added, and the
solution was stirred for 30 min more then poured into
CH.sub.2Cl.sub.2/NaHCO.sub.3(sat). The aqueous layers were further
extracted with CH.sub.2Cl.sub.2, dried (Na.sub.2SO.sub.4),
filtered, and evaporated. Column chromatography (3%
MeOH/CH.sub.2Cl.sub.2) gave pure tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(triisopropylsilyloxy)phenyl)
propylcarbamate (E7).
[0170] Tert-Butyl
2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate
(E8) was prepared from E7 according to the below:
##STR00021##
To tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(triisopropyl
silyloxy) phenyl)propylcarbamate (E7) in THF at 0.degree. C. was
added TBAF, and the solution was stirred for 45 min at 0.degree. C.
The compound was poured into EtOAc and washed with
NH.sub.4Cl.sub.(sat), dried (Na.sub.2SO.sub.4), filtered, and
evaporated. Column chromatography 6% MeOH/CH.sub.2Cl.sub.2 gave
pure tert-butyl
2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate
(E8).
[0171]
4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropa-
n-2-yl)phenylpivalate (E9) was prepared from E8 according to the
below:
##STR00022##
To tert-butyl
2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate
(E8) in pyridine was added pivaloyl chloride, and the solution was
stirred for 2 h at room temperature. The mixture was poured into
NaHCO.sub.3 and extracted with EtOAc. The organics were dried
(Na.sub.2SO.sub.4), filtered, and evaporated. Column chromatography
5% MeOH/CH.sub.2Cl.sub.2 gave pure
4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxop-
ropan-2-yl)phenylpivalate (E9).
[0172] 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl
pivalate (E10) was prepared from E9 according to the below:
##STR00023##
To
4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-
-yl)phenylpivalate (E9) in CH.sub.2Cl.sub.2 was added HCl (4N in
dioxane) and the solution was stirred for 8-10 h. The solvents were
evaporated to give pure
4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl
pivalate (E10).
[0173]
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxoprop-
an-2-yl)phenyl 1-methylcyclopropanecarboxylate (E11) was prepared
from E8 according to the below:
##STR00024##
To tert-butyl
2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate
(E8) in pyridine was added EDC, DMAP, and
1-methylcyclopropanecarboxylic acid, and the solution was flushed
with N.sub.2, capped, and stirred overnight. The mixture was poured
into EtOAc/NaHCO.sub.3(sat) and the aqueous layer was further
extracted with EtOAc. The organics were dried (Na.sub.2SO.sub.4),
filtered, and evaporated. Column chromatography 5%
MeOH/CH.sub.2Cl.sub.2 gave pure
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-y-
l)phenyl 1-methylcyclopropanecarboxylate (E11).
[0174] 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl
1-methylcyclopropanecarboxylate dihydrochloride (E12) was prepared
from E11 according to the below:
##STR00025##
To
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan--
2-yl)phenyl 1-methylcyclopropanecarboxylate (E11) in
CH.sub.2Cl.sub.2 was added HCl (4N in dioxane) and the solution was
stirred for 8-10 h. The solvents were evaporated to give pure
4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl
1-methylcyclopropanecarboxylate dihydrochloride (E12).
Examples 13-122
[0175] Using commercially available compounds and largely the
procedures set forth in Examples 2-12 and substituting the
appropriate starting materials, the compounds E13-E91 were made and
E92-E122 could be synthesized, shown in Tables 1 and 2,
respectively.
TABLE-US-00002 TABLE 1 Compounds E13-E91. ##STR00026## Example
R.sub.2 R.sub.1 10 H -i-Pr 12 H ##STR00027## 13 H Me 14 H t-Bu 15 H
--C(CH.sub.3).sub.2CH.sub.2CH.sub.3 16 H --(CH.sub.2).sub.6CH.sub.3
17 H ##STR00028## 18 H ##STR00029## 19 H ##STR00030## 20 H
##STR00031## 21 H ##STR00032## 22 H Ph 23 H 2-MePh 24 H 3-MePh 25 H
4-MePh 26 H 2,3-diMePh 27 H 2,4-diMePh 28 H 2,5-diMePh 29 H
3,4-diMePh 30 H 3,5-diMePh 31 H 2-F--Ph 32 H 3-F--Ph 33 H 4-F--Ph
34 H 2-Me, 3-F--Ph 35 H 2-Me, 4-F--Ph 36 H 2-Me, 5-F--Ph 37 H
4-t-BuPh 38 H 2-MeOPh 39 H 4-MeOPh 40 H 2,4-diMeOPh 41 H 2-MeO,
4-MePh 42 H 2-MeO-5-MePh 43 H 3,4-O--CH.sub.2--O--Ph 44 H 3-PhOPh
45 H --CH.sub.2-2-MeOPh 46 H 2-NH.sub.2--Ph 47 H 3-NH.sub.2--Ph 48
H 4-NH.sub.2--Ph 49 H 3-N(Me.sub.2)--Ph 50 H 4-N(Me.sub.2)--Ph 51 H
2-CN--Ph 52 H 4-CN--Ph 53 H 4-(CH.sub.2NH.sub.2)--Ph 54 H
2-CF.sub.3--Ph 55 H 2-pyridyl 56 H 3-pyridyl 57 H 4-pyridyl 58 H
2-Me-3-pyridyl 59 H 2-Ph--Ph 60 H 3-(COPh)--Ph 61 H ##STR00033## 62
H ##STR00034## 63 H --CH.sub.2NH.sub.2 64 H
--CH(Ph)CH.sub.2NH.sub.2 65 H ##STR00035## 66 H ##STR00036## 67 H
##STR00037## 68 H ##STR00038## 69 H --Bn 70 H
4-(CH.sub.2NMe.sub.2)--Ph 71 H ##STR00039## 72 H --CH(Me)Ph 73 H
--CH.sub.2-3,4-diMeOPh 74 H --CH.sub.2CH.sub.2Ph 75 H
--CH.sub.2CH.sub.2CH.sub.2Ph 76 H --CH.sub.2-2-MePh 77 H
--CH.sub.2-3-MePh 78 H --CH.sub.2-4-MePh 79 H ##STR00040## 80 H
##STR00041## 81 H ##STR00042## 82 H --CH2-4-FPh 83 H
--CH.sub.2CO.sub.2tBu 84 H --CHEtPh 85 H
--(CH.sub.2).sub.10CH.sub.3 86 H
--(CH.sub.2).sub.7(Z)CH.dbd.CH(CH.sub.2).sub.7CH.sub.3 87 H
##STR00043## 88 H --CH.sub.2CH.sub.2CO.sub.2Me 89 H
-(E)CH.dbd.CHCO.sub.2Me 90 H ##STR00044## 91 H -3-MeOPh
TABLE-US-00003 TABLE 2 Compounds E92-E122. ##STR00045## Example
R.sub.2 R.sub.1 92 Me Me 93 Me ##STR00046## 94 Me ##STR00047## 95
Me Ph 96 Me 2-MePh 97 Et 2,5-diMePh 98 Et 3,4-diMePh 99 Et 2-Me,
3-F--Ph 100 Et 2-Me, 4-F--Ph 101 Propyl 2-MeOPh 102 Et 2,4-diMeOPh
103 Me 3,4-O--CH.sub.2--O-- 104 Allyl 2-NH.sub.2--Ph 105 Allyl
3-NH.sub.2--Ph 106 H --CH.sub.2NH.sub.2 107 Me
--CH(Ph)CH.sub.2NH.sub.2 108 Propyl --CH(Ph)CH.sub.2NH.sub.2 109 Et
--CH(Ph)CH.sub.2NH.sub.2 110 Me Bn 111 Et Bn 112 Allyl Bn 113 Me
##STR00048## 114 Me --CH(Me)Ph 115 Et --CH(Me)Ph 116 Propyl
--CH(Me)Ph 117 Me --CH.sub.2CH.sub.2Ph 118 Et
--CH.sub.2CH.sub.2CH.sub.2Ph 119 Me --CH.sub.2-2-MePh 120 Me
--CH.sub.2-3-MePh 121 Me --CH(Et)Ph 122 Me ##STR00049##
Examples 123-131
[0176] Using commercially available compounds and largely the
procedures set forth in Examples 2-12 and substituting the
appropriate starting materials, the compounds E123-E131 were made,
shown in Table 3.
TABLE-US-00004 TABLE 3 Compounds E123-E131. ##STR00050## E123
##STR00051## E124 ##STR00052## E125 ##STR00053## E126 ##STR00054##
E127 ##STR00055## E128 ##STR00056## E129 ##STR00057## E130
##STR00058## E131
Examples 132-139
[0177] Compounds E132-E139 were prepared according to the scheme in
FIG. 7.
[0178] Methyl 2-(4-(hydroxymethyl)phenyl)acetate (E132) was
prepared according to the below:
##STR00059##
To 2-(4-(hydroxymethyl)phenyl)acetic acid in MeOH at 0.degree. C.
was added TMS-CHN.sub.2. The solution was stirred for 3 h then
quenched with a few drops of AcOH. The solvents were evaporated.
Column chromatography (SiO.sub.2, 3-15% EtOAc/Hex) gave pure methyl
2-(4-(hydroxymethyl)phenyl)acetate (E132).
[0179] Methyl 2-(4-((triisopropylsilyloxy)methyl) phenyl)acetate
(E133) was prepared from E132 according to the below:
##STR00060##
To methyl 2-(4-(hydroxymethyl)phenyl)acetate (E132) in
CH.sub.2Cl.sub.2 at 0.degree. C. was added 2,6-lutidine and
TIPS-OTf. The ice bath was removed and the solution was allowed to
warm to room temperature and stir. After 4 h the solution was
poured into NH.sub.4Cl(sat) and CH.sub.2Cl.sub.2 and the organic
layer was further extracted with NH.sub.4Cl(sat). The organics were
dried (MgSO.sub.4) filtered and evaporated. Column chromatography
(SiO.sub.2, 0-15% EtOAc/Hexanes) gave pure methyl
2-(4-((triisopropylsilyloxy)methyl)phenyl)acetate (E133).
[0180] Methyl
3-(1,3-dioxoisoindolin-2-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl)pr-
opanoate (E134) was prepared from E133 according to the below:
##STR00061##
To a solution of LiHMDS in THF cooled to -78.degree. C. was added a
cooled solution (-78.degree. C.) of methyl
2-(4-((triisopropylsilyloxy) methyl)phenyl)acetate (E133) in THF
via syringe. The solution was stirred at -78.degree. C. for 30 min.
Bromo-methyl phthalimide was added directly to the anion and the
solution stirred for 2 h at -78.degree. C. The reaction was then
poured into NH.sub.4Cl(sat) and extracted with EtOAc. The organics
were dried (MgSO.sub.4), filtered, and evaporated. Column
chromatography (SiO.sub.2, 0-20% EtOAc/Hexanes) gave pure methyl
3-(1,3-dioxoisoindolin-2-yl)-2-(4-((triisopropylsilyloxy)
methyl)phenyl)propanoate (E134).
[0181] 2-(2-carboxy-2-(4-((triisopropylsilyloxy) methyl)phenyl)
ethylcarbamoyl)benzoic acid (E135) was prepared from E134 according
to the below:
##STR00062##
To methyl
3-(1,3-dioxoisoindolin-2-yl)-2-(4-((triisopropylsilyloxy)methyl-
)phenyl)propanoate (E134) in THF/H.sub.2O was added LiOH.H.sub.2O,
and the solution was stirred for 1.5 h or until conversion to
product was visible by LC-MS. The solution was then poured into
EtOAc/NH.sub.4Cl(sat)/1 N HCl (3:1) and the aqueous layer was
further extracted with EtOAc. The organics were dried (MgSO.sub.4),
filtered, and evaporated to give crude
2-(2-carboxy-2-(4-((triisopropylsilyloxy)methyl)phenyl)ethylcarbamoyl)ben-
zoic acid (E135).
[0182]
3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-((triisoprop-
ylsilyloxy)methyl)phenyl)propanamide (E136) was prepared from E135
according to the below:
##STR00063##
To
2-(2-carboxy-2-(4-((triisopropylsilyloxy)methyl)phenyl)ethylcarbamoyl)-
benzoic acid (E135) in pyridine was added EDC, DMAP and
6-aminoisoquinoline and the solution was flushed with N.sub.2,
capped, and stirred overnight. The mixture was poured into
EtOAc/NaHCO.sub.3(sat) and the aqueous layer was further extracted
with EtOAc. The organics were dried (MgSO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, 5%
MeOH/CH.sub.2Cl.sub.2) gave pure
3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-((triisopropylsily-
loxy)methyl)phenyl)propanamide (E136).
[0183]
3-amino-N-(isoquinolin-6-yl)-2-(4-((triisopropylsilyloxy)methyl)phe-
nyl)propanamide (E137) was prepared from E136 according to the
below:
##STR00064##
To
3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-((triisopropyls-
ilyloxy)methyl)phenyl)propanamide (E136) in EtOH was added
NH.sub.2--NH.sub.2 and the solution was refluxed for 1.2-2 h. The
solids were filtered, and the solvents were evaporated. Column
chromatography (SiO.sub.2, 5% 2N NH.sub.3-MeOH/CH.sub.2Cl.sub.2)
gave pure
3-amino-N-(isoquinolin-6-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl)pr-
opanamide (E137).
[0184] Tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl-
)propylcarbamate (E138) was prepared from E137 according to the
below:
##STR00065##
To
3-amino-N-(isoquinolin-6-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl-
)propanamide (E137) in CH.sub.2Cl.sub.2 at 0.degree. C. was added a
solution of Boc.sub.2O in CH.sub.2Cl.sub.2 also cooled to 0.degree.
C. before addition. The solution was stirred for 30 min at
0.degree. C. and additional Boc.sub.2O was added and the solution
was stirred for 30 min more then poured into
CH.sub.2Cl.sub.2/NaHCO.sub.3(sat). The aqueous layers were further
extracted with CH.sub.2Cl.sub.2, dried (MgSO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, 3%
MeOH/CH.sub.2Cl.sub.2) gave tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl-
)propylcarbamate (E138).
[0185] Tert-butyl
2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamat-
e (E139).
##STR00066##
To tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl-
)propylcarbamate (E138) in THF at 0.degree. C. was added TBAF, and
the solution was stirred for 45 min at 0.degree. C. The compound
was poured into EtOAc and washed with NH.sub.4Cl(sat), dried
(MgSO.sub.4), filtered, and evaporated. Column chromatography
(SiO.sub.2, 6% MeOH/CH.sub.2Cl.sub.2) gave pure tert-butyl
2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamat-
e (E139).
Examples 140-143
[0186] Compounds E140-E143 were prepared according to the scheme in
FIG. 8.
[0187]
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxoprop-
an-2-yl)benzyl 2-methylbenzoate (E140) was prepared from E139
according to the below:
##STR00067##
To tert-butyl
2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamat-
e (E139) in pyridine was added 2-methylbenzoyl chloride and the
solution was stirred for 2 h at room temperature. The mixture was
poured into NaHCO.sub.3 and extracted with EtOAc. The organics were
dried (MgSO.sub.4), filtered, and evaporated. Column chromatography
(SiO.sub.2, 5% MeOH/CH.sub.2Cl.sub.2) gave pure
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-y-
l)benzyl 2-methylbenzoate (E140).
[0188] 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
2-methylbenzoate dihydrochloride (E141) was prepared from E140
according to the below:
##STR00068##
To
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan--
2-yl)benzyl 2-methylbenzoate (E140) in CH.sub.2Cl.sub.2 was added
HCl (4N in dioxane) and the solution was stirred for 8-10 h. The
solvents were evaporated to give pure
4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
2-methylbenzoate dihydrochloride (E141).
[0189]
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxoprop-
an-2-yl)benzyl cyclohexanecarboxylate (E142) was prepared from E139
according to the below:
##STR00069##
[0190] To tert-butyl
2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamat-
e (E139) in pyridine was added EDC, DMAP, and cyclohexanecarboxylic
acid, and the solution was flushed with N.sub.2, capped, and
stirred overnight. The mixture was poured into
EtOAc/NaHCO.sub.3(sat) and the aqueous layer was further extracted
with EtOAc. The organics were dried (MgSO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, 5%
MeOH/CH.sub.2Cl.sub.2) gave pure
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-y-
l)benzyl cyclohexanecarboxylate (E142).
[0191] 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
cyclohexanecarboxylate dihydrochloride (E143) was prepared from
E142 according to the below:
##STR00070##
To
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan--
2-yl)benzyl cyclohexanecarboxylate (E142) in CH.sub.2Cl.sub.2 was
added HCl (4 N in dioxane) and the solution was stirred for 8-10 h.
The solvents were evaporated to give pure
4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
cyclohexanecarboxylate dihydrochloride (E143).
Example 144
[0192]
3-amino-2-(4-(hydroxymethyl)phenyl)-N-(isoquinolin-6-yl)propanamide
dihydrochloride (E144) was prepared from E139 according to the
below:
##STR00071##
To tert-butyl
2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamat-
e (E139) in THF and water and cooled to 0.degree. C. was added HCl
(1 N in Et.sub.2O). After 30 min the mixture was warmed to room
temperature and the solution was stirred for 48 h. 2 M NH.sub.3 in
MeOH was added. The solvents were evaporated and the mixture
purified by column chromatography (SiO.sub.2, O-5-10% (2 M NH.sub.3
in MeOH)/CH.sub.2Cl.sub.2). The compound was dissolved in DCM/MeOH
and 1 N HCl in Et.sub.2O added. The solvents were evaporated to
give pure
3-amino-2-(4-(hydroxymethyl)phenyl)-N-(isoquinolin-6-yl)propanamide
dihydrochloride (E144).
Examples 145-148
[0193] Compounds E145-E148 were prepared according to the scheme in
FIG. 9.
[0194]
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxoprop-
an-2-yl)benzyl 2,4-dimethylbenzoate (E145) was prepared E139
according to the below:
##STR00072##
To 2,4-dimethylbenzoic acid in pyridine was added EDC, DMAP, and
tert-butyl
2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamat-
e (E139), and the solution was capped and stirred overnight. The
mixture was poured into EtOAc/NaHCO.sub.3(sat) and the aqueous
layer was further extracted with EtOAc. The organics were dried
(MgSO.sub.4), filtered, and evaporated. Column chromatography
(SiO.sub.2, 0-5% MeOH/CH.sub.2Cl.sub.2 gradient) gave pure
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-y-
l)benzyl 2,4-dimethylbenzoate (E145).
[0195] 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
2,4-dimethylbenzoate dihydrochloride (E146) was prepared from E145
according to the below:
##STR00073##
To
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan--
2-yl)benzyl 2,4-dimethylbenzoate (E145) in CH.sub.2Cl.sub.2 was
added HCl (4 N in dioxane) and the solution was stirred for 8-10 h.
The solvents were evaporated to give pure
4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
2,4-dimethylbenzoate dihydrochloride (E146).
[0196]
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxoprop-
an-2-yl)benzyl butyrate (E148) was prepared from E139 according to
the below:
##STR00074##
To butyric acid in pyridine was added EDC, DMAP, and tert-butyl
2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamat-
e (E139), and the solution was capped and stirred overnight. The
mixture was poured into EtOAc/NaHCO.sub.3(sat) and the aqueous
layer was further extracted with EtOAc. The organics were dried
(MgSO.sub.4), filtered, and evaporated. Column chromatography
(SiO.sub.2, 0-5% MeOH/CH.sub.2Cl.sub.2 gradient) gave pure
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-y-
l)benzyl butyrate (E148).
[0197] 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
butyrate dihydrochloride (E148) was prepared from E147 according to
the below:
##STR00075##
To
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan--
2-yl)benzyl butyrate (E147) in CH.sub.2Cl.sub.2 was added HCl (4 N
in dioxane) and the solution was stirred for 8-10 h The solvents
were evaporated to give pure
4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
butyrateate dihydrochloride (E148).
Examples 149-175
[0198] Using commercially available compounds and largely the
procedures set forth in Examples 140-143 and substituting the
appropriate starting materials, the compounds E149-E175 have been
made, shown in Table 4.
TABLE-US-00005 TABLE 4 Compounds E149-E175. ##STR00076## Example R
149 ##STR00077## 150 -iPr 151 -tBu 152 --(CH.sub.2).sub.6CH.sub.3
153 ##STR00078## 154 ##STR00079## 155 ##STR00080## 156 --Ph 157
--Bn 158 --CH.sub.2CH.sub.2Ph 159 --CH.sub.2--OPh 160 ##STR00081##
161 3,5-diMePh 162 ##STR00082## 163 --(CH.sub.2).sub.10CH.sub.3 164
##STR00083## 165 3-MeOPh 166 4-MeOPh 167 2,4-diOMePh 168
3,4-O--CH.sub.2--O--Ph 169 ##STR00084## 170 --CHPh.sub.2 171
2-Ph--Ph 172 ##STR00085## 173 ##STR00086## 174 ##STR00087## 175
##STR00088##
Examples 176-196
[0199] Using commercially available compounds and largely the
procedures set forth in Examples 140-143 and substituting the
appropriate starting materials, the compounds E176-E196, could be
made, shown in Table 5.
TABLE-US-00006 TABLE 5 Compounds E176-E196. ##STR00089## Example R
176 2-MeOPh 177 4-NHMePh 178 4-NMe.sub.2Ph 179 4-OEtPh 180 3-MePh
181 4-MePh 182 2,3-diMePh 183 2,6-MePh 184 3,4-MePh 185
##STR00090## 186 2-ClPh 187 3-ClPh 188 4-ClPh 189 2-FPh 190 3-FPh
191 4-FPh 192 2,4-diClPh 193 2,4-diFPh 194 ##STR00091## 195
##STR00092## 196 ##STR00093##
Example 197
[0200]
(S)-4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxo-
propan-2-yl)benzyl 2,4-dimethylbenzoate (E145-S) and
(R)-4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-
-2-yl)benzyl 2,4-dimethylbenzoate (E145-R) were prepared from E145
according to the scheme in FIG. 10.
4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-y-
l)benzyl 2,4-dimethylbenzoate was dissolve in methanol and the R
and S enantiomers separated by supercritical fluid chromatography
(Chiralpak AS-H column, eluent: 18.8% MeOH, 0.2%
dimethylethylamine, 80% CO.sub.2). The enantiomers were then each
purified by column chromotagraphy (SiO.sub.2, 0-5%
MeOH/CH.sub.2Cl.sub.2 gradient). The enantiomeric excess for each
enantiomer was >98%.
[0201]
(S)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
2,4-dimethylbenzoate dihydrochloride (E197-S) was prepared from
E145-S according to the scheme in FIG. 6. To
(S)-4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-
-2-yl)benzyl 2,4-dimethylbenzoate (E145-S) in CH.sub.2Cl.sub.2 was
added HCl (4 N in dioxane) and the solution was stirred for 8-10 h.
The solvents were evaporated to give pure
(S)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
2,4-dimethylbenzoate dihydrochloride (E197-S). Analysis by chiral
HPLC (Chiralpak AS-H, eluent: 90:10:0.1 EtOH:H.sub.2O:diethylamine)
showed enantiomeric excess >98%.
[0202]
(R)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
2,4-dimethylbenzoate dihydrochloride (E197-R) was prepared from
E145-R according to the scheme in FIG. 6. To
(S)-4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-
-2-yl)benzyl 2,4-dimethylbenzoate (E145-R) in CH.sub.2Cl.sub.2 was
added HCl (4 N in dioxane) and the solution was stirred for 8-10 h.
The solvents were evaporated to give pure
(R)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
2,4-dimethylbenzoate dihydrochloride (E197-R). Analysis by chiral
HPLC (Chiralpak AS-H, eluent: 90:10:0.1 EtOH:H.sub.2O:diethylamine)
showed enantiomeric excess >98%.
Examples 198-203
[0203] Compounds E199-E203 were prepared according to the scheme in
FIG. 11.
[0204] (4-iodobenzyloxy)triisopropylsilane (E199) was prepared from
E198 according to the below:
##STR00094##
To a solution of (4-iodophenyl)methanol (E198) and imidazole in
CH.sub.2Cl.sub.2 at 0.degree. C. was added dropwise TIPSCl. The
reaction mixture was stirred overnight. The solution was quenched
with H.sub.2O and the CH.sub.2Cl.sub.2 layer separated. The organic
layer was further washed with 0.5N HCl and NaHCO.sub.3(sat). The
combined organic layers were dried (MgSO.sub.4), filtered, and
evaporated. The crude yellow oil,
(4-iodobenzyloxy)triisopropylsilane (E199), was used directly in
the next step.
[0205] Ethyl
2-cyano-2-(4-((triisopropylsilyloxy)methyl)phenyl)acetate (E200)
was prepared from E199 according to the below:
##STR00095##
To a solution of ethyl 2-cyanoacetate and
(4-iodobenzyloxy)triisopropylsilane (E199) in dioxane were added
Cs.sub.2CO.sub.3, CuI, and picolinic acid. The mixture was stirred
overnight at 90.degree. C. The solid was removed by filtration and
the dioxane concentrated under reduced pressure. Column
chromatography (SiO.sub.2, hexane:ethyl acetate 25:1) gave pure
ethyl 2-cyano-2-(4-((triisopropylsilyloxy)methyl)phenyl)acetate
(E200) as a yellow oil.
[0206] Ethyl
3-amino-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E201)
was prepared from E200 according to the below:
##STR00096##
To a suspension of CoCl.sub.2.6H.sub.2O in THF was added ethyl
2-cyano-2-(4-((triisopropylsilyloxy)methyl)phenyl)acetate (E200).
The mixture was cooled to 0.degree. C. and NaBH.sub.4 was added to
the mixture in several portions over 30 min. The mixture was
stirred at room temperature for 4 h. The reaction was quenched with
water. The mixture was filtered and the filtrate extracted twice
with ether. The organics were dried (MgSO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, DCM:EtOH=50:1) gave
pure ethyl
3-amino-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E201)
as a yellow oil.
[0207] Ethyl
3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)pr-
opanoate (E202) was prepared from E201 according to the below:
##STR00097##
To a solution of ethyl
3-amino-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E201)
in DCM was added (Boc).sub.2O and triethylamine. The mixture was
stirred for 2 h, then washed with 0.5 N HCl and NaHCO.sub.3(sat).
The organic layer was dried (MgSO.sub.4) and concentrated to give
ethyl
3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)pr-
opanoate (E202).
[0208]
3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phe-
nyl)propanoic acid (E203) was prepared from E202 according to the
below:
##STR00098##
To a solution of ethyl
3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)pr-
opanoate (E202) in methanol was added dropwise 4 N NaOH. The
mixture was stirred for 2 h, adjusted the pH to 7 with 2 N HCl, and
extracted with ethyl acetate. The combined organic layers were
washed with 0.5 N HCl and brine, dried (MgSO.sub.4), and
concentrated in vacuo to afford
3-(tert-butoxycarbonylamino)-2-(4-((tnisopropylsilyloxy)methyl)phenyl)pro-
panoic acid (E203) as white solid.
Examples 204-206
[0209] Compounds E204-E206 were prepared according to the scheme in
FIG. 12, which is a modified procedure by Cheung, S. T. et al. Can.
J. Chem. 1977, 55, 906-910.
[0210]
3-(tert-butoxycarbonyl(methyl)amino)-2-(4-((triisopropylsilyloxy)me-
thyl)phenyl)propanoic acid (E204) was prepared from E203 according
to the below:
##STR00099##
To
3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl-
)propanoic acid (E203) in THF under N.sub.2 and cooled to 0.degree.
C. was added CH.sub.3I followed by NaH and the solution was warmed
and allowed to stir for 18 h. The mixture was taken up in EtOAc and
extracted with NH.sub.4Cl.sub.(sat), dried (MgSO.sub.4), filtered,
and evaporated. Column chromatography (SiO.sub.2, 0-10%
MeOH/CH.sub.2Cl.sub.2 gradient) gave pure
3-(tert-butoxycarbonyl(methyl)amino)-2-(4-((tnisopropylsilyloxy-
)methyl)phenyl)propanoic acid (E204).
[0211] Tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl-
)propyl(methyl)carbamate (E205) was prepared from E204 according to
the below:
##STR00100##
To
3-(tert-butoxycarbonyl(methyl)amino)-2-(4-((triisopropylsilyloxy)methy-
l)phenyl)propanoic acid (E204) in pyridine was added was added EDC,
DMAP, and 6-aminoisoquinoline, and the solution was stirred
overnight at room temperature. The mixture was poured into
NaHCO.sub.3(sat) and extracted with EtOAc. The organics were dried
(MgSO.sub.4), filtered, and evaporated. Column chromatography
(SiO.sub.2, 0-6% MeOH/CH.sub.2Cl.sub.2 gradient) gave pure
tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl-
)propyl(methyl)carbamate (E205).
[0212] Tert-butyl
2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropyl(methyl)-
carbamate (E205-1) was prepared from E205 according to the
below:
##STR00101##
To tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((thisopropylsilyloxy)methyl)phenyl)-
Propyl(methyl)carbamate (E205) in THF under N.sub.2 at 0.degree. C.
was added TBAF, and the solution was stirred for 30 min at
0.degree. C. The reaction was warmed to room temperature and
stirred another 4.5 h. The compound was poured into EtOAc and
washed with NH.sub.4Cl.sub.(sat), dried (MgSO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, 0-20%
MeOH/CH.sub.2Cl.sub.2 gradient) gave pure tert-butyl
2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropyl(methyl)-
carbamate (E205-1)
[0213]
4-(3-(tert-butoxycarbonyl(methyl)amino)-1-(isoquinolin-6-ylamino)-1-
-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E205-2) was prepared
from E205-1 according to the below:
##STR00102##
To tert-butyl
2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropyl(methyl)-
carbamate (E205-1) in pyridine was added was added EDC, DMAP, and
2,4-dimethylbenzoic acid, and the solution was stirred overnight at
room temperature. The mixture was poured into NaHCO.sub.3(sat) and
extracted with EtOAc. The organics were dried (MgSO.sub.4),
filtered, and evaporated. Column chromatography (SiO.sub.2, 0-5%
MeOH/CH.sub.2Cl.sub.2 gradient) gave pure
4-(3-(tert-butoxycarbonyl(methyl)amino)-1-(isoquinolin-6-ylamino)-1-oxopr-
opan-2-yl)benzyl 2,4-dimethylbenzoate (E205-2).
[0214]
4-(1-(isoquinolin-6-ylamino)-3-(methylamino)-1-oxopropan-2-yl)benzy-
l 2,4-dimethylbenzoate (E206) was prepared from E205-2 according to
the below:
##STR00103##
To
4-(3-(tert-butoxycarbonyl(methyl)amino)-1-(isoquinolin-6-ylamino)-1-ox-
opropan-2-yl)benzyl 2,4-dimethylbenzoate (E205-2) in
CH.sub.2Cl.sub.2 was added HCl (4 N in dioxane) and the solution
was stirred for 8-10 h. The solvents were evaporated to give pure
4-(1-(isoquinolin-6-ylamino)-3-(methylamino)-1-oxopropan-2-yl)benzyl
2,4-dimethylbenzoate (E206).
Examples 207-211
[0215] Using commercially available compounds and largely the
procedures set forth in Examples 204-206 and substituting the
appropriate starting materials, the compounds E206-E211 have been
made, shown in Table 6.
TABLE-US-00007 TABLE 6 Compounds E206-E211. ##STR00104## E207
##STR00105## E208 ##STR00106## E209 ##STR00107## E210 ##STR00108##
E211
Example 212
[0216]
3-(tert-butoxycarbonylamino)-2-(3-((triisopropylsilyloxy)methyl)phe-
nyl)propanoic acid (E212) was prepared.
##STR00109##
Using commercially available compounds and largely the procedures
set forth in Examples 198-203 and substituting the appropriate
starting materials
3-(tert-butoxycarbonylamino)-2-(3-((triisopropylsilyloxy)methyl-
)phenyl)propanoic acid (E212) was made.
Examples 213-216
[0217] Using commercially available compounds and largely the
procedures set forth in Examples 204-206 and substituting the
appropriate starting materials, the compounds E213-E216 have been
made, shown in Table 7.
TABLE-US-00008 TABLE 7 Compounds E213-E216. ##STR00110## E213
##STR00111## E214 ##STR00112## E215 ##STR00113## E216
Examples 217-225
[0218] Using commercially available compounds and largely the
procedures set forth in Examples 198-203 and Examples 204-206 and
substituting the appropriate starting materials, the compounds
E217-E225 could be made, shown in Table 8.
TABLE-US-00009 TABLE 8 Compounds E217-E225 ##STR00114## E217
##STR00115## E218 ##STR00116## E219 ##STR00117## E220 ##STR00118##
E221 ##STR00119## E222 ##STR00120## E223 ##STR00121## E224
##STR00122## E225
Example 226
[0219] 3-(isopropylamino)-N-(isoquinolin-6-yl)-2-phenylpropanamide
dihydrochloride (E226) was prepared as shown below:
##STR00123##
To 3-amino-N-(isoquinolin-6-yl)-2-phenylpropanamide in MeOH/AcOH
was added acetone and NaCNBH.sub.3. Then after 15 min the mixture
was poured into NaHCO.sub.3(sat) and extracted with
CH.sub.2Cl.sub.2. The organics were dried (Na.sub.2SO.sub.4),
filtered, and evaporated. Column chromatography (SiO.sub.2, 5% 2 N
NH.sub.3-MeOH/CH.sub.2Cl.sub.2) gave pure
3-(isopropylamino)-N-(isoquinolin-6-yl)-2-phenylpropanamide. The
compound was taken up in CH.sub.2Cl.sub.2 and HCl (1 M in
Et.sub.2O) was added. The solution was evaporated to give
3-(isopropylamino)-N-(isoquinolin-6-yl)-2-phenylpropanamide
dihydrochloride (E226).
Examples 227-230
[0220] Using commercially available compounds and largely the
procedures set forth in Example 226 and substituting the
appropriate starting materials, the compounds E227-E230 could be
made, shown in Table 9.
TABLE-US-00010 TABLE 9 Compounds E227-E230. ##STR00124## E227
##STR00125## E228 ##STR00126## E229 ##STR00127## E230
##STR00128##
Examples 231-241
[0221] Compounds E231-E241 were prepared according to the scheme in
FIG. 13.
[0222] Methyl
2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)acetate
(E232) was prepared from E231 according to the below:
##STR00129##
To 2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)acetic
acid (E231) in MeOH at 0.degree. C. was added TMS-CH.sub.2N.sub.2
until the solution persisted in a yellow color and TLC indicated
completion of the reaction. The solution stirred for 30 min and
then was quenched with a few drops of AcOH. The solvents were
evaporated and column chromatography (SiO.sub.2, 0-15%
EtOAc/Hexanes) gave pure methyl
2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)acetate
(E232).
[0223] Methyl
2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3-dioxoisoin-
dolin-2-yl)propanoate (E233) was prepared from E232 according to
the below:
##STR00130##
To a solution of LiHMDS in THF cooled to -78.degree. C. was added a
cooled solution (approx -78.degree. C.) of methyl
2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)acetate
(E232) in THF via syringe. The solution was stirred at -78.degree.
C. for 30 min. Bromo-methylphthalimide was added directly to the
anion, and the solution was immediately removed from the
-78.degree. C. bath and placed in an ice bath and stirred for 2 h.
The reaction was then poured into NH.sub.4Cl.sub.(sat) and
extracted with EtOAc. The organics were dried (Na.sub.2SO.sub.4),
filtered, and evaporated. Column chromatography (SiO.sub.2, 15-20%
EtOAc/Hexanes) gave pure methyl
2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3-dioxoisoin-
dolin-2-yl)propanoate (E233).
[0224]
2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-2-carboxy-
ethylcarbamoyl)benzoic acid (E234) was prepared from E233 according
to the below:
##STR00131##
To methyl
2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3--
dioxoisoindolin-2-yl)propanoate (E233) in THF/H.sub.2O was added
LiOH*H.sub.2O, and the solution was stirred for 1.5 h or until
complete conversion to product was visible by LC-MS. The solution
was then poured into EtOAc/NH.sub.4Cl(sat)/1 N HCl (3:1) and the
aqueous layer was further extracted with EtOAc. The organics were
dried (Na.sub.2SO.sub.4), filtered, evaporated, and dried to give
crude
2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-2-carboxyethylc-
arbamoyl)benzoic acid (E234).
[0225]
2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3-di-
oxoisoindolin-2-yl)-N-(isoquinolin-6-yl)propanamide (E235) was
prepared from E234 according to the below:
##STR00132##
[0226] To
2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-2-carb-
oxyethylcarbamoyl)benzoic acid (E234) in pyridine was added EDC,
DMAP, and 6-aminoisoquinoline, and the solution was flushed with
N.sub.2, capped, and stirred overnight. The mixture was poured into
EtOAc/NaHCO.sub.3(sat) and the aqueous layer was further extracted
with EtOAc. The organics were dried (Na.sub.2SO.sub.4), filtered,
and evaporated. Column chromatography (SiO.sub.2, 4%
MeOH/CH.sub.2Cl.sub.2) gave pure
2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3-dioxoiso-
indolin-2-yl)-N-(isoquinolin-6-yl)propanamide (E235).
[0227]
3-amino-2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-N-(-
isoquinolin-6-yl)propanamide (E236) was prepared from E235
according to the below:
##STR00133##
To
2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3-dioxo-
isoindolin-2-yl)-N-(isoquinolin-6-yl)propanamide (E235) in EtOH was
added NH.sub.2--NH.sub.2 and the solution was stirred for 7 h at
room temperature then heated to 50.degree. C. for 1 h. The solution
was cooled, the solids were filtered, and the solvents were
evaporated. Column chromatography (SiO.sub.2, 5-8%2 N
NH.sub.3-MeOH/CH.sub.2Cl.sub.2) gave pure
3-amino-2-(5-((tert-butyldimethylsilyloxy)
methyl)thiophen-2-yl)-N-(isoquinolin-6-yl)propanamide (E236).
[0228] Tert-butyl
2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(isoquinolin-6--
ylamino)-3-oxopropylcarbamate (E237) was prepared from E236
according to the below:
##STR00134##
To 3-amino-2-(5-((tert-butyldimethylsilyloxy)
methyl)thiophen-2-yl)-N-(isoquinolin-6-yl)propanamide (E236) in
CH.sub.2Cl.sub.2 at 0.degree. C. was added a solution of Boc.sub.2O
in CH.sub.2Cl.sub.2 (also cooled to 0.degree. C. before addition).
The solution was stirred at 0.degree. C. for 2 h and then poured
into CH.sub.2Cl.sub.2 and NaHCO.sub.3(sat). The solution was
further extracted with CH.sub.2Cl.sub.2 and the combined organics
were dried (Na.sub.2SO.sub.4), filtered, and evaporated. Column
chromatography (SiO.sub.2, 3% MeOH/CH.sub.2Cl.sub.2) gave pure
tert-butyl
2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(isoquinolin-6--
ylamino)-3-oxopropylcarbamate (E237).
[0229] Tert-butyl
2-(5-(hydroxymethyl)thiophen-2-yl)-3-(isoquinolin-6-ylamino)-3-oxopropylc-
arbamate (E238) was prepared from E237 according to the below:
##STR00135##
To tert-butyl
2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(isoquinolin-6--
ylamino)-3-oxopropylcarbamate (E237) in THF at 0.degree. C. was
added TBAF and the solution was stirred for 0.degree. C. for 30 min
then warmed to room temperature for 2 h. The compound was poured
into EtOAc and washed with NH.sub.4Cl.sub.(sat), dried
(Na.sub.2SO.sub.4), filtered, and evaporated. Column chromatography
(SiO.sub.2, 6% MeOH/CH.sub.2Cl.sub.2) gave pure tert-butyl
2-(5-(hydroxymethyl)thiophen-2-yl)-3-(isoquinolin-6-ylamino)-3-oxopropylc-
arbamate (E238).
[0230]
3-amino-2-(5-(hydroxymethyl)thiophen-2-yl)-N-(isoquinolin-6-yl)prop-
anamide dihydrochloride (E239) was prepared from E238 according to
the below:
##STR00136##
To a solution of tert-butyl
2-(5-(hydroxymethyl)thiophen-2-yl)-3-(isoquinolin-6-ylamino)-3-oxopropylc-
arbamate (E238) in CH.sub.2Cl.sub.2 was added 4 N HCl-dioxane and
the solution was stirred for 4 h. The solvents were evaporated to
give
3-amino-2-(5-(hydroxymethyl)thiophen-2-yl)-N-(isoquinolin-6-yl)propanamid-
e dihydrochloride (E239).
[0231]
(5-(3-tert-butoxylcarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopro-
pan-2-yl)methyl 2,4 dimethylbenzoate (E240) was prepared from E239
according to the below:
##STR00137##
To tert-butyl
2-(5-(hydroxymethyl)thiophen-2-yl)-3-(isoquinolin-6-ylamino)-3-oxopropylc-
arbamate (E238) in pyridine was added EDC, DMAP, and 2,4-dimethyl
benzoic acid, and the solution was flushed with N.sub.2, capped,
and stirred overnight. The mixture was poured into
EtOAc/NaHCO.sub.3(sat) and the aqueous layer was further extracted
with EtOAc. The organics were dried (Na.sub.2SO.sub.4), filtered,
and evaporated. Column chromatography (SiO.sub.2, 4%
MeOH/CH.sub.2Cl.sub.2) gave pure
(5-(3-tert-butoxylcarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2--
yl)methyl 2,4 dimethylbenzoate (E240).
[0232]
(5-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)thiophen-2-y-
l)methyl 2,4-dimethylbenzoate dihydrochloride (E241) was prepared
from E240 according to the below:
##STR00138##
To
(5-(3-tert-butoxylcarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-
-2-yl)methyl 2,4 dimethylbenzoate (E240) in CH.sub.2Cl.sub.2 was
added HCl (4 N in dioxane) and the solution was stirred overnight.
The solvents were evaporated to give pure
(5-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)thiophen-2-yl)meth-
yl 2,4-dimethylbenzoate dihydrochloride (E241).
Examples 242-248
[0233] Using commercially available compounds and largely the
procedures set forth in Examples 231-241 and substituting the
appropriate starting materials, E242-E248 could be synthesized,
shown in Table 10.
TABLE-US-00011 TABLE 10 Compounds E242-E248. ##STR00139## Example
R.sup.1 R.sup.2 242 --CH.sub.2Ph H 243 -3,5-diMePh H 244
##STR00140## Me 245 ##STR00141## H 246 --(CH.sub.2).sub.2CH.sub.3
Me 247 i-Pr H 248 --Ph Me
Examples 249-253
[0234] Methyl
2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan--
2-yl)phenoxy)acetate (E250) was prepared from E249 according to the
scheme in FIG. 14. To tert-butyl
2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate
(E249) in DMF cooled to -35.degree. C. was added NaH, and the
solution was stirred at -35.degree. C. for 30 min. Then, methyl
bromoacetate was added and the solution was warmed and stirred at
0.degree. C. for 1 h. The solution was poured into
NaHCO.sub.3(sat)/EtOAc and further extracted with EtOAc. The
combined organics were dried (Na.sub.2SO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, 3-4%
MeOH/CH.sub.2Cl.sub.2) gave pure methyl
2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan--
2-yl)phenoxy)acetate (E250).
[0235]
2-(4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopr-
opan-2-yl)phenoxy)acetic acid (E251) was prepared from E250
according to the below:
##STR00142##
To methyl
2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1--
oxopropan-2-yl)phenoxy)acetate (E250) in THF/H.sub.2O/MeOH at
0.degree. C. was added LiOH*H.sub.2O, and the solution was stirred
for 2 h at 0.degree. C. The mixture was then quenched with HCl (1
N, Et.sub.2O) and evaporated. Column chromatography (SiO.sub.2, 20%
MeOH/CH.sub.2Cl.sub.2) gave pure
2-(4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-o-
xopropan-2-yl)phenoxy)acetic acid (E251).
[0236] 2,4-dimethylphenyl
2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan--
2-yl)phenoxy)acetate (E252) was prepared from E251 according to the
below:
##STR00143##
To
2-(4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropa-
n-2-yl)phenoxy)acetic acid (E251) in pyridine was added EDC, DMAP,
and 2,4-dimethylphenol, and the solution was stirred for 5 h. The
mixture was then poured into EtOAc/NaHCO.sub.3(sat) and extracted
with EtOAc. The combined organics were dried (Na.sub.2SO.sub.4),
filtered, and evaporated. Column chromatography (SiO.sub.2, 2-3%
MeOH/CH.sub.2Cl.sub.2) gave 2,4-dimethylphenyl
2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan--
2-yl)phenoxy)acetate (E252).
[0237] 2,4-dimethylphenyl
2-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetate
dihydrochloride (E253) was prepared from E252 according to the
below:
##STR00144##
To 2,4-dimethylphenyl
2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan--
2-yl)phenoxy)acetate (E252) in CH.sub.2Cl.sub.2 was added HCl (4 N,
dioxane) and the solution was stirred overnight. The solvents were
evaporated to give 2,4-dimethylphenyl
2-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetate
dihydrochloride (E253).
Examples 254-273
[0238] Using commercially available compounds and largely the
procedures set forth in Examples 249-253 and substituting the
appropriate starting materials E254-E261 (shown in Table 11) were
made and E262-E273 (shown in Table 12) could be synthesized.
TABLE-US-00012 TABLE 11 Compounds E254-E261. ##STR00145## Example X
R 254 O Me 255 O H 256 O -2,4-diMePh 257 O -i-Pr 258 O --CH.sub.2Ph
259 O -3,5-diMePh 260 NH Ph 261 NH --(CH.sub.2).sub.3CH.sub.3
TABLE-US-00013 TABLE 12 Compounds E262-E273. ##STR00146## Example X
R.sup.1 R.sup.2 262 O Ph H 263 O 4-MeOPh Me 264 O 2,4-di-F--Ph Me
265 O --CH.sub.2Ph H 266 O --CH.sub.2CH.dbd.CH.sub.2 H 267 O
##STR00147## Me 268 NH 2,4-diMePh Me 269 NH 3,5-diMePh Me 270 NH
2-F--Ph H 271 NH --CH.sub.2-4-MeOPh Me 272 NH -2-MeOPh H 273 NH
-3-pyridyl H
Example 274
[0239] Using commercially available compounds and largely the
procedures set forth in Examples 249-253 and substituting the
appropriate starting materials E274 was made.
##STR00148##
Examples 275-278
[0240] Compounds E275-E278 were prepared according to the scheme
presented in FIG. 15. Tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(2-oxo-2-phenylethoxy)phenyl)
propylcarbamate (E275) was prepared from E249 according to the
below:
##STR00149##
To tert-butyl
2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate
(E249) in DMF cooled to -35.degree. C. was added NaH and the
solution was stirred at -35.degree. C. for 30 min. Then,
2-bromoacetophenone was added and the solution was warmed and
stirred at 0.degree. C. for 2 h. The solution was poured into
NaHCO.sub.3(sat)/EtOAc and further extracted with EtOAc. The
combined organics were dried (Na.sub.2SO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, 3%
MeOH/CH.sub.2Cl.sub.2) gave tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(2-oxo-2-phenylethoxy)phenyl)
propylcarbamate (E275).
[0241]
3-amino-N-(isoquinolin-6-yl)-2-(4-(2-oxo-2-phenylethoxy)phenyl)prop-
anamide dihydrochloride (E277) was prepared from E275 according to
the below:
##STR00150##
To tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(2-oxo-2-phenylethoxy)phenyl)
propylcarbamate (E275) in CH.sub.2Cl.sub.2 was added HCl (4 N,
dioxane) and the solution was stirred overnight. The solvents were
evaporated to give pure
3-amino-N-(isoquinolin-6-yl)-2-(4-(2-oxo-2-phenylethoxy)phenyl)-
propanamide dihydrochloride (E277).
[0242] Tert-butyl
2-(4-(2-hydroxy-2-phenylethoxy)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopro-
pylcarbamate (E276) was prepared from E275 according to the
below:
##STR00151##
To tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(2-oxo-2-phenylethoxy)phenyl)
propylcarbamate (E275) in EtOH was added NaBH.sub.4 and the
solution was stirred for 20 min at room temperature. The mixture
was then poured into NaHCO.sub.3(sat) and extracted with
CH.sub.2Cl.sub.2. The combined organics were dried
(Na.sub.2SO.sub.4), filtered, and evaporated. Column chromatography
(SiO.sub.2, 5% MeOH/CH.sub.2Cl.sub.2) gave pure tert-butyl
2-(4-(2-hydroxy-2-phenylethoxy)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopro-
pylcarbamate (E276).
[0243]
3-amino-2-(4-(2-hydroxy-2-phenylethoxy)phenyl)-N-(isoquinolin-6-yl)-
propanamide dihydrochloride (E278) was prepared from E276 according
to the below:
##STR00152##
To tert-butyl
2-(4-(2-hydroxy-2-phenylethoxy)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopro-
pylcarbamate (E276) in CH.sub.2Cl.sub.2 was added HCl (4 N,
dioxane) and the solution was stirred overnight. The solvents were
evaporated to give
3-amino-2-(4-(2-hydroxy-2-phenylethoxy)phenyl)-N-(isoquinolin-6-yl)propan-
amide dihydrochloride (E278).
Examples 279-288
[0244] Using commercially available compounds and largely the
procedures set forth in Examples 275-278 and substituting the
appropriate starting materials E279-E282 (shown in Table 13) were
made and E283-E288 (shown in Table 14) could be synthesized.
TABLE-US-00014 TABLE 13 Compounds E279-E282. ##STR00153## Example X
R 279 O Ph 280 OH Ph 281 O -4-MeOPh 282 O -2-MeOPh
TABLE-US-00015 TABLE 14 Compounds E283-E288. ##STR00154## Example X
R1 R2 283 O -2-F--Ph Me 284 O -2,4 diCl--Ph H 285 O -3-MePh H 286
OH -4-MeOPh H 287 OH -2-MeOPh H 288 OH -3-MePh Me
Examples 289-290
[0245] Compounds E289 and E290 were prepared according to the
scheme presented in FIG. 16.
[0246] Tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(phenethoxyphenyl)
propylcarbamate (E289) was prepared from E249 according to the
below:
##STR00155##
To tert-butyl
2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate
(E249) in DMF cooled to -35.degree. C. was added NaH and the
solution was stirred at -40.degree. C. for 30 min. Then,
2-bromoethylbenzene was added and the solution was warmed and
stirred at room temperature for 2 h. The solution was poured into
NaHCO.sub.3(sat)/EtOAc and further extracted with EtOAc. The
combined organics were dried (Na.sub.2SO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, 3-4%
MeOH/CH.sub.2Cl.sub.2) gave pure tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(phenethoxyphenyl)
propylcarbamate (E289).
[0247]
3-amino-N-(isoquinolin-6-yl)-2-(4-phenethoxyphenyl)propanamide
dihydrochloride (E290) was prepared from E289 according to the
below:
##STR00156##
To tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(phenethoxyphenyl)
propylcarbamate (E289) in CH.sub.2Cl.sub.2 was added HCl (4 N,
dioxane) and the solution was stirred overnight. The solvents were
evaporated to give
3-amino-N-(isoquinolin-6-yl)-2-(4-phenethoxyphenyl)propanamide
dihydrochloride (E290).
Examples 291-299
[0248] Using commercially available compounds and largely the
procedures set forth in Examples 289-290 and substituting the
appropriate starting materials E291-E292 (Table 15) were made and
E294-E299 (Table 16) could be synthesized.
TABLE-US-00016 TABLE 15 Compounds E291-E292. ##STR00157## Example R
291 --(CH.sub.2).sub.2Ph 292 --CH.sub.2Ph** ** E292 was synthesized
from previous schemes carried out in which the benzyl was in place
of the TIPS protecting group.
[0249] Using commercially available compounds and largely the
procedures set forth in Examples 289-290 and substituting the
appropriate starting materials E293 was made.
##STR00158##
TABLE-US-00017 TABLE 16 Compounds E294-E299. ##STR00159## Example
R1 R2 294 --CH.sub.2 -4-F--Ph H 295 --CH.sub.2-2-MePh Me 296
--CH.sub.2-2-CNPh Me 297 --(CH.sub.2).sub.2-4-MePh H 298
--(CH.sub.2).sub.2-2-FPh H 299 ##STR00160## H
Examples 300-308
[0250] Compounds E300-E308 were prepared according to the scheme in
FIG. 17.
[0251] Benzyl 2-(4-hydroxyphenyl)acetate (E301) was prepared from
E300 according to the below:
##STR00161##
To 2-(4-hydroxyphenyl)acetic acid in DMF cooled to 0.degree. C. was
added K.sub.2CO.sub.3 and the solution was stirred for 30 min.
Then, benzyl bromide was added and the solution stirred at
0.degree. C. and was allowed to slowly warm to 15-20.degree. C.
After all the ice was melted the solution was poured into
NH.sub.4Cl.sub.(sat) and extracted with EtOAc. The combined
organics were dried (Na.sub.2SO.sub.4), filtered, and evaporated.
Column chromatography (SiO.sub.2, 0-35% EtOAc/Hex) gave pure benzyl
2-(4-hydroxyphenyl)acetate (E301).
[0252] Benzyl 2-(4-(triisopropylsiloxy)phenyl)acetate (E302) was
prepared from E301 according to the below:
##STR00162##
To benzyl 2-(4-hydroxyphenyl)acetate (E301) in CH.sub.2Cl.sub.2 at
0.degree. C. was added 2,6-lutidine and TIPS-OTf and the solution
stirred for 2.5 h at 0.degree. C. The mixture was poured into
NH.sub.4Cl.sub.(sat) and extracted with CH.sub.2Cl.sub.2. The
combined organics were dried (Na.sub.2SO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, 0-15% EtOAc/Hex) gave
pure benzyl 2-(4-(triisopropylsiloxy)phenyl)acetate (E302).
[0253] Benzyl 3-cyano-2-(triisopropylsilyloxy)phenyl)propanoate
(E303) was prepared from E302 according to the below:
##STR00163##
To a solution of LiHMDS in THF at -78.degree. C. was added a
solution of benzyl 2-(4-(triisopropylsiloxy)phenyl)acetate (E302)
in THF also cooled to approx -78.degree. C., and this mixture was
allowed to stir at -78.degree. C. for 30 min. Iodoacetonitrile was
then added and the mixture was warmed to 0.degree. C. and stirred
for 2 h. The mixture was poured into NH.sub.4Cl.sub.(sat) and
extracted with EtOAc. The combined organics were dried
(Na.sub.2SO.sub.4), filtered, and evaporated. Column chromatography
(SiO.sub.2, 0-25% EtOAc/Hex) gave pure benzyl
3-cyano-2-(triisopropylsilyloxy)phenyl)propanoate (E303).
[0254] Benzyl
4-(tertbutoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)butanoate
(E304) was prepared from E303 according to the below:
##STR00164##
[0255] To a solution of benzyl
3-cyano-2-(triisopropylsilyloxy)phenyl)propanoate (E303) in MeOH
cooled to 0.degree. C. was added CoCl.sub.2*6H.sub.2O and
NaBH.sub.4 and the solution was allowed to stir for 20 min. Then,
HCl (1.25 N in MeOH) was added and the solution stirred an
additional 20 min at 0.degree. C. The solvents were evaporated and
the mixture was taken up in CH.sub.2Cl.sub.2 and cooled to
0.degree. C. Boc.sub.2O and NEt.sub.3 were added and the solution
stirred at 0.degree. C. for 1.5 h. The mixture was poured into
NH.sub.4Cl.sub.(sat) and extracted with CH.sub.2Cl.sub.2. The
combined organics were dried (Na.sub.2SO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, 10-20% EtOAc/Hexanes)
gave pure benzyl
4-(tertbutoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)butanoate
(E304).
[0256] Preparation of
4-(tertbutoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)butanoic
acid (E305) was prepared from E304 according to the below:
##STR00165##
To benzyl
4-(tert-butoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)-
butanoate (E304) in EtOAc was added Pd/C (10%) and the solution was
kept under a H.sub.2 atomosphere for 2 h. The mixture was filtered
over Celite and the solvent was evaporated to give
4-(tert-butoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)butanoic
acid (E305).
[0257] Tert-butyl
4-(isoquinolin-6-ylamino)-4-oxo-3-(4-(triisopropylsilyloxy)phenyl)butylca-
rbamate (E306) was prepared from E305 according to the below:
##STR00166##
To
4-(tert-butoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)butanoi-
c acid (E305) in pyridine was added EDC, DMAP, and 6-AlQ, and the
solution was stirred at room temperature overnight. The mixture was
poured into NaHCO.sub.3(sat) and extracted with EtOAc. The combined
organics were dried (Na.sub.2SO.sub.4), filtered, and evaporated.
Column chromatography (SiO.sub.2, 4% MeOH/CH.sub.2Cl.sub.2) gave
pure tert-butyl
4-(isoquinolin-6-ylamino)-4-oxo-3-(4-(triisopropylsilyloxy)phenyl)butylca-
rbamate (E306).
[0258] Preparation of tert-butyl
3-(4-hydroxyphenyl)-4-(isoquinolin-6-ylamino)-4-oxobutylcarbamate
(E307) was prepared from E306 according to the following:
##STR00167##
To tert-butyl
4-(isoquinolin-6-ylamino)-4-oxo-3-(4-(triisopropylsilyloxy)phenyl)butylca-
rbamate (E306) in THF at 0.degree. C. was added TBAF and the
solution was stirred at 0.degree. C. for 30 min. The solution was
poured into NH.sub.4Cl.sub.(sat) and extracted with EtOAc. The
combined organics were dried (Na.sub.2SO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, 5-8%
MeOH/CH.sub.2Cl.sub.2) gave pure tert-butyl
3-(4-hydroxyphenyl)-4-(isoquinolin-6-ylamino)-4-oxobutylcarbamate
(E307).
[0259] Preparation of
4-amino-2-(4-hydroxyphenyl)-N-(isoquinolin-6-yl)butanamide
dihydrochloride (E308) was prepared from E307 according to the
below:
##STR00168##
To tert-butyl
3-(4-hydroxyphenyl)-4-(isoquinolin-6-ylamino)-4-oxobutylcarbamate
(E307) in CH.sub.2Cl.sub.2 was added HCl (4 N in dioxane) and 2
drops of H.sub.2O and the solution was stirred overnight at room
temperature. The solvents were evaporated to give
4-amino-2-(4-hydroxyphenyl)-N-(isoquinolin-6-yl)butanamide
dihydrochloride (E308).
Examples 309-318
[0260] Using commercially available compounds and largely the
procedures set forth in this application and substituting the
appropriate starting materials E309-E318 could be synthesized.
TABLE-US-00018 TABLE 17 Compounds E309-E318. ##STR00169## Example
R1 R2 309 H H 310 --CO--Ph Me 311 --CO-2,4-diMePh H 312
--COCH.sub.2Ph H 313 --CO(CH.sub.2).sub.3CH.sub.3 H 314
--CH.sub.2COPh Me 315 --CH.sub.2CO-4-MeOPh Me 316
--CH.sub.2--CH(OH)--Ph H 317 --CH.sub.2-3-MeOPh H 318
--(CH.sub.2).sub.2Ph Me
Examples 319-325
[0261] Compounds E319-E325 were prepared according to the scheme in
FIG. 18.
[0262] Preparation of methyl
3-(diphenylmethyleneamino)-2-phenylpropanoate (E320) was prepared
from E319 according to the below:
##STR00170##
To methyl 3-amino-2-phenylpropanoate hydrochloride in
CH.sub.2Cl.sub.2 was added benzophenone imine, and the solution was
stirred overnight at room temperature. The mixture was then washed
with H.sub.2O and the organics were dried (Na.sub.2SO.sub.4),
filtered, and evaporated. Column chromatography (SiO.sub.2, 5-20%
EtOAc/Hexanes) gave pure methyl
3-(diphenylmethyleneamino)-2-phenylpropanoate (E320).
[0263] Methyl
3-(diphenylmethyleneamino)-2-methyl-2-phenylpropanoate (E321) was
prepared from E320 according to the below:
##STR00171##
[0264] To a solution of LiHMDS in THF cooled to -78.degree. C. was
added a solution of methyl
3-(diphenylmethyleneamino)-2-phenylpropanoate (E320) in THF also
cooled to approximately -78.degree. C. This solution stirred for 30
min at -78.degree. C., then methyl iodide was added directly and
the solution was warmed to 0.degree. C. After 3 h the solution was
poured into NH.sub.4Cl.sub.(sat) and extracted with EtOAc. The
combined organics were dried (Na.sub.2SO.sub.4) filtered, and
evaporated. Column chromatography (SiO.sub.2, O-15% EtOAc/Hexanes)
gave pure methyl
3-(diphenylmethyleneamino)-2-methyl-2-phenylpropanoate (E321).
[0265] 3-amino-2-methyl-2-phenylpropanoic acid hydrochloride (E322)
was prepared from E321 according to the below:
##STR00172##
A mixture of methyl
3-(diphenylmethyleneamino)-2-methyl-2-phenylpropanoate (E321) and 6
N HCl was refluxed overnight. The solution was cooled and
evaporated to give 3-amino-2-methyl-2-phenylpropanoic acid
hydrochloride (E322).
[0266] 3-(tert-butoxycarbonylamino)-2-methyl-2-phenylpropanoic acid
(E323) was prepared from E322 according to the below:
##STR00173##
To a solution of Boc.sub.2O in dioxane cooled to 0.degree. C. was
added a solution of 3-amino-2-methyl-2-phenylpropanoic acid
hydrochloride (E322) in 1 N NaOH and this solution stirred 3 h and
the solution was then washed with
NaHCO.sub.3(sat)/CH.sub.2Cl.sub.2. The aqueous layer was acidified
with HCl (1 N) and extracted with CH.sub.2Cl.sub.2. These combined
organics were dried (Na.sub.2SO.sub.4), filtered, and evaporated to
give 3-(tert-butoxycarbonylamino)-2-methyl-2-phenylpropanoic acid
(E323).
[0267] Tert-butyl
3-isoquinolin-6-yl)-2-methyl-3-oxo-2-phenylpropylcarbamate (E324)
was prepared from E323 according to the below:
##STR00174##
To 3-(tert-butoxycarbonylamino)-2-methyl-2-phenylpropanoic acid
(E323) in pyridine was added EDC, DMAP, and 6-AlQ, and solution was
stirred at room temperature for 48 h. The mixture was poured into
NaHCO.sub.3(sat) and extracted with EtOAc. The combined organics
were dried (Na.sub.2SO.sub.4), filtered, and evaporated. Column
chromatography (SiO.sub.2, 3% MeOH/CH.sub.2Cl.sub.2) gave pure
tert-butyl
3-isoquinolin-6-yl)-2-methyl-3-oxo-2-phenylpropylcarbamate
(E324).
[0268] 3-amino-N-(isoquinolin-6-yl)-2-methyl-2-phenylpropanamide
dihydrochloride (E325) was prepared from E324 according to the
below:
##STR00175##
To tert-butyl
3-isoquinolin-6-yl)-2-methyl-3-oxo-2-phenylpropylcarbamate (E324)
in CH.sub.2Cl.sub.2 was added HCl (4 N in dioxane) and the solution
was stirred overnight at room temperature. The solvents were
evaporated to give
3-amino-N-(isoquinolin-6-yl)-2-methyl-2-phenylpropanamide
dihydrochloride (E325).
Examples 326-334
[0269] Using commercially available compounds and largely the
procedures set forth in the Examples above and substituting the
appropriate starting materials, E326-E334 could be synthesized,
shown in Table 18.
TABLE-US-00019 TABLE 18 Compounds E326-E334. ##STR00176## Example X
n R.sup.1 R.sup.2 326 --OH 1 Me Me 327 --CH.sub.2OH 1 Me H 328
--OCOPh 2 Me H 329 --OCO-2,4-diMePh 1 --CH.sub.2Ph H 330
--OCOCH.sub.2Ph 1 --CH.sub.2Ph H 331 --CH.sub.2OCO-3,5-diMePh 1 Me
H 332 --CH.sub.2OCO-2,4-diMePh 1 --CH.sub.2-4-MeOPh Me 333
--CH.sub.2OCO--(CH.sub.2).sub.2CH.sub.3 1 --CH.sub.2-2-MeOPh H 334
--CH.sub.2OCO-2,4-diMePh 2 Me H
Examples 335-338
[0270] 2-fluoro-4-nitrobenzamide (E335) was prepared according to
the below:
##STR00177##
To 2-fluoro-4-nitrobenzoic acid suspended in CH.sub.2Cl.sub.2 under
Ar was added DMF then oxalyl chloride. The reaction was stirred at
room temperature 1.5 h then the solvent was evaporated. The residue
was dissolved in THF and ammonia gas was bubbled through the
reaction for 15 min. The solvent was evaporated and the residue
partitioned between EtOAc and water. The aqueous layer was
extracted with EtOAc. The extracts were dried (MgSO.sub.4),
filtered, and evaporated. Column chromatography (SiO.sub.2, 0-100%
EtOAc/Hex) gave pure 2-fluoro-4-nitrobenzamide (E335).
[0271] 4-amino-2-fluorobenzamide (E336) was prepared from E335
according to the below:
##STR00178##
2-fluoro-4-nitrobenzamide (E335) was dissolved in EtOH under Ar and
10% Pd/C added. The reaction was pump-purged with H.sub.2 and left
stirring at room temperature overnight. The catalyst was removed by
filtration and the reaction concentrated to give pure
4-amino-2-fluorobenzamide (E336).
[0272] Tert-butyl
3-(4-carbamoyl-3-fluorophenylamino)-3-oxo-2-(4-((triisopropylsilyloxy)met-
hyl)phenyl)propylcarbamate (E337) was prepared from E336 according
to the below:
##STR00179##
To
3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl-
)propanoic acid in pyridine was added EDC, DMAP, and
4-amino-2-fluorobenzamide (E336), and the solution was stirred
overnight at room temperature. The mixture was poured into
NaHCO.sub.3(sat) and extracted with EtOAc. The extracts were dried
(MgSO.sub.4), filtered, and evaporated. Column chromatography
(SiO.sub.2, 0-6% MeOH/CH.sub.2Cl.sub.2 gradient) gave pure
tert-butyl
3-(4-carbamoyl-3-fluorophenylamino)-3-oxo-2-(4-((triisopropylsilyloxy)met-
hyl)phenyl)propylcarbamate (E337).
[0273] Tert-butyl
3-(4-carbamoyl-3-fluorophenylamino)-2-(4-(hydroxymethyl)phenyl)-3-oxoprop-
ylcarbamate (E337-1) was prepared from E337 according to the
below:
##STR00180##
To tert-butyl
3-(4-carbamoyl-3-fluorophenylamino)-3-oxo-2-(4-((thisopropylsilyloxy)meth-
yl)phenyl)propylcarbamate (E337) in THF under N.sub.2 at 0.degree.
C. was added TBAF, and the solution was stirred for 30 min at
0.degree. C. The reaction was warmed to room temperature and
stirred another 3.5 h. The compound was poured into EtOAc and
washed with NH.sub.4Cl.sub.(sat), dried (MgSO.sub.4), filtered, and
evaporated. Column chromatography (SiO.sub.2, 0-20%
MeOH/CH.sub.2Cl.sub.2 gradient) gave pure tert-butyl
3-(4-carbamoyl-3-fluorophenylamino)-2-(4-(hydroxymethyl)phenyl)-3-oxoprop-
ylcarbamate (E337-1)
[0274]
4-(3-(tert-butoxycarbonylamino)-1-(4-carbamoyl-3-fluorophenylamino)-
-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E337-2) was prepared
from E337-1 according to the below:
##STR00181##
To tert-butyl
3-(4-carbamoyl-3-fluorophenylamino)-2-(4-(hydroxymethyl)phenyl)-3-oxoprop-
ylcarbamate (E337-1) in pyridine was added was added EDC, DMAP, and
2,4-dimethylbenzoic acid, and the solution was stirred overnight at
room temperature. The mixture was poured into NaHCO.sub.3(sat) and
extracted with EtOAc. The organics were dried (MgSO.sub.4),
filtered, and evaporated. Column chromatography (SiO.sub.2, O-5%
MeOH/CH.sub.2Cl.sub.2 gradient) gave pure
4-(3-(tert-butoxycarbonylamino)-1-(4-carbamoyl-3-fluorophenylamino)-1-oxo-
propan-2-yl)benzyl 2,4-dimethylbenzoate (E337-2).
[0275]
4-(3-amino-1-(4-carbamoyl-3-fluorophenylamino)-1-oxopropan-2-yl)ben-
zyl 2,4-dimethylbenzoate (E338) was prepared from E337-2 according
to the below:
##STR00182##
[0276] To
4-(3-(tert-butoxycarbonylamino)-1-(4-carbamoyl-3-fluorophenylami-
no)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E337-2) in
CH.sub.2Cl.sub.2 was added HCl (4 N in dioxane) and the solution
was stirred overnight. The solvents were evaporated to give pure
4-(3-amino-1-(4-carbamoyl-3-fluorophenylamino)-1-oxopropan-2-yl)benzyl
2,4-di methyl benzoate (E338).
Examples 339-370
[0277] Using commercially available compounds and largely the
procedures set forth in Examples 335-338 and substituting the
appropriate starting materials, the compounds E339-E354 (Table 19)
and E355-E370 (Table 20) could be made.
TABLE-US-00020 TABLE 19 Compounds E339-E354. ##STR00183## Example
R1 R2 R3 R4 R5 339 H Bu H H F 340 Me Bu H H F 341 H Ph H H H 342 Me
Ph H H H 343 H 3,5-diMePh F H H 344 H 2,4-diMePh H F H 345 H Bn H H
F 346 H cyclohexyl Me H H 347 Me cyclopentyl H Me H 348 Me 3-MePh H
H Me 349 H 4-MePh H H H 350 H 3-thienyl H H H 351 Me 2,4-diFPh H H
H 352 H 3,5-diClPh H H H 353 Me 2-thienyl H H H 354 H 4-MeOPh H H
H
TABLE-US-00021 TABLE 20 Compounds E355-E370. ##STR00184## Example
R1 R2 R3 R4 R5 355 H Bu H H F 356 Me Bu H H F 357 H Ph H H H 358 Me
Ph H H H 359 H 3,5-diMePh F H H 360 H 2,4-diMePh H F H 361 H Bn H H
F 362 H cyclohexyl Me H H 363 Me cyclopentyl H Me H 364 Me 3-MePh H
H Me 365 H 4-MePh H H H 366 H 3-thienyl H H H 367 Me 2,4-diFPh H H
H 368 H 3,5-diClPh H H H 369 Me 2-thienyl H H H 370 H 4-MeOPh H H
H
Examples 371-377
[0278] Compounds E371-E377 were prepared according to the scheme in
FIG. 19.
[0279] For the preparation of methyl
3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)pr-
opanoate (E372), to a 0.degree. C. solution of methyl
3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)pr-
opanoic acid (E371) in MeOH was added a 2.0 M solution of
trimethylsilyldiazomethane in hexanes. The solution was stirred for
20 min at room temperature and then quenched by the addition of a
few drops of AcOH. The solution was concentrated and the residue,
methyl
3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)pr-
opanoate (E372), was used without purification.
[0280] For the preparation of methyl
3-(tert-butoxycarbonylamino)-2-(4-(hydroxymethyl)phenyl)propanoate
(E373), to a 0.degree. C. solution of methyl
3-(tert-butoxycarbonylamino)-2-(4-((tnisopropylsilyloxy)methyl)phenyl)pro-
panoate (E372) in THF was added a 1 M solution of
tetrabutylammonium fluoride in THF, and the reaction was stirred
overnight at room temperature. The reaction was quenched with
saturated aqueous NH.sub.4Cl, and extracted with EtOAc (3.times.).
The combined organics were washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated. The residue was purified by
silica gel column chromatography (eluting with 0% to 50%
EtOAc/hexanes) to yield methyl
3-(tert-butoxycarbonylamino)-2-(4-(hydroxymethyl)phenyl)propanoate
(E373).
[0281] For the preparation of
4-(3-(tert-butoxycarbonylamino)-1-methoxy-1-oxopropan-2-yl)benzyl
2,4-dimethylbenzoate (E374, R=2,4-Me.sub.2Ph), to a solution of
methyl
3-(tert-butoxycarbonylamino)-2-(4-(hydroxymethyl)phenyl)propanoate
(E373) in pyridine was added
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride
(EDCI), 4-(dimethylamino)pyridine (DMAP), and 2,4-dimethylbenzoic
acid. The reaction was stirred overnight at room temperature. After
addition of EtOAc and saturated aqueous NaHCO.sub.3, the mixture
was extracted with EtOAc (3.times.). The combined organics were
washed with brine, dried over Na.sub.2SO.sub.4, and concentrated.
The residue was purified by silica gel column chromatography
(eluting with 20% to 80% EtOAc/hexanes) to yield
4-(3-(tert-butoxycarbonylamino)-1-methoxy-1-oxopropan-2-yl)benzyl
2,4-dimethylbenzoate (E374, R=2,4-Me.sub.2Ph).
[0282] For the preparation of
3-(tert-butoxycarbonylamino)-2-(4-((2,4-dimethylbenzoyloxy)methyl)phenyl)-
propanoic acid (E375, R=2,4-Me.sub.2Ph), to a solution of
4-(3-(tert-butoxycarbonylamino)-1-methoxy-1-oxopropan-2-yl)benzyl
2,4-dimethylbenzoate (E374, R=2,4-Me.sub.2Ph) in 2:1 THF/H.sub.2O
was added LiOH.H.sub.2O and the solution was stirred at room
temperature for 3 h. After addition of 1 N HCl (until the pH was
acidic), the mixture was extracted with EtOAc (3.times.). The
combined organics were washed with brine, dried over
Na.sub.2SO.sub.4, filtered, and concentrated to yield
3-(tert-butoxycarbonylamino)-2-(4-((2,4-dimethylbenzoyloxy)methyl)phenyl)-
propanoic acid (E375, R=2,4-Me.sub.2Ph).
[0283] For the preparation of
4-(3-(tert-butoxycarbonylamino)-1-(1-methoxyisoquinolin-6-ylamino)-1-oxop-
ropan-2-yl)benzyl 2,4-dimethylbenzoate (E376, R=2,4-Me.sub.2Ph,
X=1-OMe), to a solution of
3-(tert-butoxycarbonylamino)-2-(4-((2,4-dimethylbenzoyloxy)methyl)phenyl)-
propanoic acid (E375, R=2,4-Me.sub.2Ph) in pyridine was added EDCI,
DMAP, and 6-amino-1-methoxyisoquinoline. The solution was stirred
overnight at room temperature. The mixture was diluted with EtOAc
and saturated aq. NaHCO.sub.3 solution. The mixture was extracted
with EtOAc (3.times.). The combined organics were washed with
brine, dried over Na.sub.2SO.sub.4, and concentrated. The residue
was purified by silica gel column chromatography (eluting with 0%
to 80% EtOAc/hexanes) to yield
4-(3-(tert-butoxycarbonylamino)-1-(1-methoxyisoquinolin-6-ylamino)-1-oxop-
ropan-2-yl)benzyl 2,4-dimethylbenzoate (E376, R=2,4-Me.sub.2Ph,
X=1-OMe).
[0284] For the preparation of
4-(3-amino-1-(1-methoxyisoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
2,4-di methyl benzoate (E377, R=2,4-Me.sub.2Ph, X=1-OMe), to a
solution of
4-(3-(tert-butoxycarbonylamino)-1-(1-methoxyisoquinolin-6-ylamino)-1-o-
xopropan-2-yl)benzyl 2,4-dimethylbenzoate (E376, R=2,4-Me.sub.2Ph,
X=3-Me) in CH.sub.2Cl.sub.2 was added 4N HCl in dioxane and the
solution was stirred overnight at room temperature. The solution
was concentrated. The residue was diluted with dichloromethane and
concentrated again to yield
4-(3-amino-1-(1-methoxyisoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl
2,4-dimethylbenzoate (E377, R=2,4-Me.sub.2Ph, X=2-OMe) as the
hydrochloride salt.
Examples 378-380
[0285] Using largely the procedures shown above, the following
compounds E378-E380 were synthesized, shown in Table 21.
TABLE-US-00022 TABLE 21 Compounds E378-E380. ##STR00185## Example R
378 ##STR00186## 379 ##STR00187## 380 ##STR00188##
Examples 381-397
[0286] Using largely the procedures shown above, the following
compounds E381-E397 could be synthesized, shown in Table 22.
TABLE-US-00023 TABLE 22 Compounds E381-E397. ##STR00189## Example R
381 ##STR00190## 382 ##STR00191## 383 ##STR00192## 384 ##STR00193##
385 ##STR00194## 386 ##STR00195## 387 ##STR00196## 388 ##STR00197##
389 ##STR00198## 390 ##STR00199## 391 ##STR00200## 392 ##STR00201##
393 ##STR00202## 394 ##STR00203## 395 ##STR00204## 396 ##STR00205##
397 ##STR00206##
Examples 398-404
[0287] Compounds E399-E404 were prepared according to the scheme in
FIG. 20.
[0288] Methyl 2-phenyl-3-(triisopropylsilyloxy)propanoate (E399)
was prepared from E398 according to the below:
##STR00207##
To methyl 3-hydroxy-2-phenylpropanolate in CH.sub.2Cl.sub.2 was at
0.degree. C. was added 2,6-lutidine and TIPS-OTf, and this solution
was stirred for 2 h at room temperature. The mixture was poured
into NH.sub.4Cl.sub.(sat) and extracted with CH.sub.2Cl.sub.2. The
organics were dried (Na.sub.2SO.sub.4), filtered, and evaporated.
Column chromatography 0-10% EtOAc/Hex gave pure methyl
2-phenyl-3-(triisopropylsilyloxy)propanoate (E399).
[0289] 2-phenyl-3-(triisopropylsilyloxy)propanoic acid (E400) was
prepared from E399 according to the below:
##STR00208##
To methyl 2-phenyl-3-(triisopropylsilyloxy)propanoate (E399) in
THF/H.sub.2O/MeOH was added LiOH*H.sub.2O and the solution was stir
at room temperature overnight. The solution was poured into
NH.sub.4Cl.sub.(sat)/HCl (1 N) (3:1) and extracted with EtOAc. The
combined organics were dried (Na.sub.2SO.sub.4), filtered, and
evaporated. Column chromatography (0%-4% MeOH/CH.sub.2Cl.sub.2)
gave pure 2-phenyl-3-(triisopropylsilyloxy)propanoic acid
(E400).
[0290]
N-(isoquinolin-6-yl)-2-phenyl-3-(triisopropylsilyloxy)propanamide
(E401) was prepared from E400 according to the below:
##STR00209##
To 2-phenyl-3-(triisopropylsilyloxy)propanoic acid (E400) in
pyridine was added EDC, DMAP, and 6-aminoisoquinoline, and the
solution was flushed with N.sub.2, capped, and stirred overnight.
The mixture was poured into NaHCO.sub.3(sat) and extracted with
EtOAC. The combined organics were dried (Na.sub.2SO.sub.4),
filtered, and evaporated. Column chromatography (3-4%
MeOH/CH.sub.2Cl.sub.2) gave pure
N-(isoquinolin-6-yl)-2-phenyl-3-(triisopropylsilyloxy)propanamide
(E401).
[0291] 3-hydroxy-N-(isoquinolin-6-yl)-2-phenylpropanamide (E402)
was prepared from E401 according to the below:
##STR00210##
To
N-(isoquinolin-6-yl)-2-phenyl-3-(triisopropylsilyloxy)propanamide
(E401) in THF cooled to 0.degree. C. was added TBAF and this
solution was stirred for 3 h at 0.degree. C. The mixture was poured
into EtOAc/NH.sub.4Cl.sub.(sat) and washed with
NH.sub.4Cl.sub.(sat). The organics were dried (Na.sub.2SO.sub.4),
filtered, and evaporated. Column chromatography (0-10%
MeOH/CH.sub.2Cl.sub.2) gave pure
3-hydroxy-N-(isoquinolin-6-yl)-2-phenylpropanamide (E402).
[0292] 3-(isoquinolin-6-ylamino)-3-oxo-2-phenylpropyl
methanesulfonate (E403) was prepared from E402 according to the
below:
##STR00211##
To 3-hydroxy-N-(isoquinolin-6-yl)-2-phenylpropanamide (E402) in
pyridine at 0.degree. C. was added MsCl, and this solution was
stirred at 0.degree. C. for 2.5 h. The mixture was poured into
NaHCO.sub.3(sat) and extracted with EtOAc. The combined organics
were dried (Na.sub.2SO.sub.4), filtered, and evaporated to give
3-(isoquinolin-6-ylamino)-3-oxo-2-phenylpropyl methanesulfonate
(E403).
[0293]
N-(isoquinolin-6-yl)-3-(4-methylpiperazin-1-yl)-2-phenylpropanamide
(E404) was prepared from E403 according to the below:
##STR00212##
To 3-(isoquinolin-6-ylamino)-3-oxo-2-phenylpropyl methanesulfonate
(E403) in methanol was added 1-methypiperazine, and the solution
was stirred overnight at 50.degree. C. The solvents were evaporated
and column chromatography 10-20% 2 N NH.sub.3-MeOH/CH.sub.2Cl.sub.2
gave
N-(isoquinolin-6-yl)-3-(4-methylpiperazin-1-yl)-2-phenylpropanamide
(E404).
Examples 405-428
[0294] Using commercially available compounds and largely the
procedures set forth in the previous examples and substituting the
appropriate starting materials, the compounds E405-E410 (Table 23)
and E411-E428 (Table 24 and Table 25) could be synthesized.
TABLE-US-00024 TABLE 23 Compounds E405-E410. ##STR00213##
4-(1-(isoquinolin-6-ylamino)-1-oxo-3-(4-
phenylpiperazin-1-yl)propan-2-yl)phenyl 2- phenylacetate 405
##STR00214## 4-(3-(4-acetylpiperazin-1-yl)-1-(isoquinolin-6-
ylamino)-1-oxopropan-2-yl)phenyl benzoate 406 ##STR00215##
4-(3-(4-(cyclopropylmethyl)piperazin-1-yl)-1-
(isoquinolin-6-ylamino)-1-oxopropan-2- yl)phenyl
3,5-dimethylbenzoate 407 ##STR00216##
3-(1,4-diazepan-1-yl)-N-(isoquinolin-6-yl)-2-(4-
(2-oxo-2-phenylethoxy)phenyl)propanamide 408 ##STR00217##
4-(3-(4-benzyl-1,4-diazepan-1-yl)-1-
(isoquinolin-6-ylamino)-1-oxopropan-2- yl)phenyl 2-phenylacetate
409 ##STR00218## 4-(1-(isoquinolin-6-ylamino)-1-oxo-3-(piperidin-
1-yl)propan-2-yl)phenyl 2-phenylpropanoate 410
TABLE-US-00025 TABLE 24 Compounds E411-E419. ##STR00219## Example
R.sup.1 R.sup.2 R.sup.3 n 411 Me Me CO-2,4diMePh 1 412 Me Me
CO--CH2Ph 1 413 Me CH.sub.2-4-HOPh CO--(CH.sub.2).sub.2CH.sub.3 1
414 Me CH.sub.2-2-HOPh CH.sub.2--COPh 1 415 Me CH.sub.2-4-FPh
CH.sub.2CO-4-MeOPh 1 416 Et CH.sub.2--Ph CH.sub.2C(OH)-2-MeOPh 2
417 Et Me CH.sub.2CH.sub.2Ph 2 418 Me CH.sub.2-3-pyridyl COBn 2 419
Me CH.sub.2-4-pyridyl COPh 2
TABLE-US-00026 TABLE 25 Compounds E420-E428. ##STR00220## Example
R.sup.1 R.sup.2 R.sup.3 n 420 Me Me CO-2,4diMePh 1 421 Me Me
##STR00221## 1 422 Me CH.sub.2-4-MeOPh CO--(CH2)2CH3 2 423 Me
CH.sub.2-2-HOPh CO-4-MePh 1 424 Me CH.sub.2-3-FPh COPh 1 425 Et
CH2--Ph CO-3,5-diMePh 2 426 Et Me CO--Bn 1 427 Me ##STR00222##
CO(CH.sub.2).sub.2CH.sub.3 2 428 Me CH.sub.2-4-pyridyl
CO(CH.sub.2).sub.2Ph 1
Examples 429-433
[0295] Compounds E429-E433 were prepared according to the scheme in
FIG. 21, which is a modified procedure of Calmes et al., Eur. J.
Org. Chem. 2000, 2459-2466.
[0296] Methyl
3-(1,3-dioxoisoindolin-2-yl)-2-(thiophen-3-yl)propanoate (E429) was
prepared according to the below:
##STR00223##
To pure methyl 2-(thiophen-3-yl)acetate in THF cooled to
-78.degree. C. was added LiHMDS and the solution stirred at
-78.degree. C. for 30 min. Then N-(bromomethyl)phthalimide was
added directly and the solution was allowed to warm to 0.degree. C.
The mixture was poured into NaHCO.sub.3(sat) extracted with EtOAc,
dried (Na.sub.2SO.sub.4), filtered, and evaporated. Column
chromatography (SiO.sub.2, 0-40% EtOAc/Hex) gave pure methyl
3-(1,3-dioxoisoindolin-2-yl)-2-(thiophen-3-yl)propanoate
(E429).
[0297] 3-amino-2-(thiophen-3-yl)propanoic acid hydrochloride (E430)
was prepared from E429 according to the below:
##STR00224##
To methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(thiophen-3-yl)propanoate
(E429) was added 6 N HCl and the solution was refluxed for 4 h. The
solvents were evaporated to give 3-amino-2-(thiophen-3-yl)propanoic
acid (E430).
[0298] 3-(tert-butoxycarbonylamino)-2-(thiophen-3-yl) propanoic
acid (E431) was prepared from E430 according to the below:
##STR00225##
To Boc.sub.2O in dioxane at 0.degree. C. was added a cooled
solution (0.degree. C.) of 3-amino-2-(thiophen-3-yl)propanoic acid
hydrochloride (E430) in 1 N NaOH. The solution was stirred at
0.degree. C. for 30 min, then at room temperature for 4 h. The
mixture was acidified with HCl and extracted with EtOAc and
NH.sub.4Cl.sub.(sat). The organics were dried (Na.sub.2SO.sub.4),
filtered, and evaporated to give pure of
3-(tert-butoxycarbonylamino)-2-(thiophen-3-yl)propanoic acid
(E431).
[0299] Tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(thiophen-3-yl)propylcarbamate
(E432) was prepared from E431 according to the below:
##STR00226##
To 3-(tert-butoxycarbonylamino)-2-(thiophen-3-yl)propanoic acid
(E431) in pyridine was added was added EDC, DMAP, and
6-aminoisoquinoline, and the solution was stirred overnight at room
temperature. The mixture was poured into NaHCO.sub.3(sat) and
extracted with EtOAc. The organics were dried (Na.sub.2SO.sub.4),
filtered, and evaporated. Column chromatography (SiO.sub.2, 3%
MeOH/CH.sub.2Cl.sub.2) gave pure tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(thiophen-3-yl) propylcarbamate
(E432).
[0300] 3-amino-N-(isoquinolin-6-yl)-2-(thiophen-3-yl)propanamide
dihydrochloride (E433) was prepared from E432 according to the
below:
##STR00227##
To tert-butyl
3-(isoquinolin-6-ylamino)-3-oxo-2-(thiophen-3-yl)propylcarbamate
(E432) in CH.sub.2Cl.sub.2 was added HCl (4 N in dioxane) and the
solution was stirred for 8-10 h. The solvents were evaporated to
give pure 3-amino-N-(isoquinolin-6-yl)-2-(thiophen-3-yl)propanamide
dihydrochloride (E433).
Examples 434-456
[0301] Using commercially available compounds and largely the
procedures set forth in Examples 429-433 and substituting the
appropriate starting materials, the following compounds E434-E441
(Table 26) and E442-E456 (Table 27) were made.
TABLE-US-00027 TABLE 26 Compounds E434-E441. ##STR00228## Example X
R.sub.4 R.sub.2 R.sub.1 434 H (.+-.)-3-thienyl Me Me 435 H
(.+-.)-3-thienyl H H 436 H C.sub.6H.sub.5 H H 437 H C.sub.6H.sub.5
Me Me 438 F C.sub.6H.sub.5 H H 439 F C.sub.6H.sub.5 Me Me 440 H
(.+-.)-2-thienyl H H 441 Cl (.+-.)-2-thienyl Me Me
TABLE-US-00028 TABLE 27 Compounds E442-E456. ##STR00229## Example X
R.sub.4 R.sub.2 R.sub.1 442 H (R)-C.sub.6H.sub.5 H H 443 H
(S)-C.sub.6H.sub.5 H H 444 OH p-fluoro-C.sub.6H.sub.4 Me Me 445 H
p-fluoro-C.sub.6H.sub.4 benzyl H 446 H Benzyl Me H 447 H p-fluoro
benzyl Me H 448 OH 3-pyridyl H H 449 H 4-pyridyl Me Me 450 OH
3-furyl H H 451 H cyclopropyl Me Me 452 H cyclopentyl Me Me 453 OH
cyclohexyl H H 454 H 3-benzo[b]thiophene Me Me 455 H ##STR00230## H
H 456 OH 2-oxazole H H
Example 457
[0302] Topical pharmaceutical compositions for lowering intraocular
pressure are prepared by conventional methods and formulated as
follows:
TABLE-US-00029 Ingredient Amount (wt %) Active ingredient 0.50
Dextran 70 0.1 Hydroxypropyl methylcellulose 0.3 Sodium chloride
0.77 Potassium chloride 0.12 Disodium EDTA 0.05 Benzalkonium
chloride 0.01 HCl and/or NaOH pH 5.5-6.5 Purified water q.s. to
100%
[0303] A compound according to this invention is used as the active
ingredient. When the composition is topically administered to the
eyes once daily, the above composition decreases intraocular
pressure in a patient suffering from glaucoma.
Example 458
[0304] Example 457 is repeated using E6 according to this
invention. When administered as a drop 2 times per day, the above
composition substantially decreases intraocular pressure and serves
as a neuroprotective agent.
Example 459
[0305] Example 457 is repeated using a gamma amino acid isoquinolyl
amide according to this invention. When administered as a drop
twice per day, the above composition substantially decreases
intraocular pressure.
Example 460
[0306] Example 457 is repeated using a benzamide according to this
invention. When administered as a drop twice per day, the above
composition substantially decreases allergic symptoms and relieves
dry eye syndrome.
Example 461
[0307] Example 457 is repeated using E19 according to this
invention. When administered as a drop as needed, the above
composition substantially decreases hyperemia, redness, and ocular
irritation.
Example 462
[0308] Example 457 is repeated using E18 according to this
invention. When administered as a drop 4 times per day, the above
composition substantially decreases intraocular pressure and serves
as a neuroprotective agent.
Example 463
[0309] Example 457 is repeated using E21 according to this
invention. When administered as a drop twice per day, the above
composition substantially decreases intraocular pressure.
Example 464
[0310] Example 457 is repeated using E115 according to this
invention. When administered as a drop twice per day, the above
composition substantially decreases ocular pressure, allergic
symptoms, and relieves dry eye syndrome.
Reference Example 2
Cell-Based Porcine Trabecular Meshwork (PTM) Assay
[0311] The anterior section of porcine eyes are harvested within 4
h post-mortem. The iris and ciliary body are removed and trabecular
meshwork cells are harvested by blunt dissection. Finely minced
trabecular meshwork tissue are plated into collagen-coated 6-well
plates in Medium-199 containing 20% fetal bovine serum (FBS). After
two passages at confluence, cells are transferred to low-glucose
DMEM containing 10% FBS. Cells are used between passage 3 and
passage 8.
[0312] Cells are plated into fibronectin-coated, glass multiwell
plates the day before compound testing under standard culture
conditions. Compounds are added to cells in the presence of 1%
FBS-containing DMEM and 1% DMSO. When compounds are incubated with
the cells for the duration determined to be optimal, the media and
compound is removed and cells fixed for 20 min in 3% methanol-free
paraformaldehyde. Cells are rinsed twice with phosphate buffered
saline (PBS) and cells are permeabilized with 0.5% Triton X-100 for
two min. Following an additional two washes with PBS, F-actin is
stained with Alexa-fluor 488-labelled phalloidin and nuclei are
stained with DAPI.
[0313] Data is reduced to the mean straight actin-fiber length and
normalized to DMSO-treated control cells (100%) and 50 .mu.M
Y-27632 (0%). Y-27632 is a rho-kinase inhibitor known to result in
the depolymerization of F-actin in these cells.
Reference Example 3
Norepinephrine Transporter (NET) Membrane Radioligand Binding
Assays
[0314] Total cell membranes are prepared from MDCK cells expressing
the recombinant human norepinehrine transporter (hNET) grown to
confluence in 150 mm tissue culture dishes. Cells are scraped into
standard medium and pelleted at 1600 g. The medium is discarded and
the pellet resuspended in 5 mL per plate of ice-cold binding buffer
(100 mM NaCl, 50 mM Tris, pH 7.4 at room temperature) by
trituration, and the cells are repelleted at 20,000 g. Supernatant
is discarded and cells are resuspended in binding buffer (50 mM
Tris-HCl, pH 7.4, 100 mM NaCl, 1 .mu.M leupeptin, 10 .mu.M PMSF)
and homogenized with a polytron (Brinkman) at 25,000 revs/min for 5
s. Centrifugation, resuspension, and homogenization are repeated
and a sample of suspension is used for Bradford protein
determination (BioRad). Samples of membrane suspensions are frozen
at -80.degree. C. prior to use. Typical yields are about 100 .mu.g
membrane protein per 106 cells. Assays performed in duplicate are
initiated with 0.2 nM [125I]RTI-55. Non-specific binding is
determined by the inclusion of 10 .mu.M desipramine. Incubation is
carried out for 3 h at 4.degree. C. Assays are terminated by rapid
filtration over GF/B glass-fiber filters soaked in 0.5%
polyethylineimine using an automated cell harvester (Brandel)
followed by three rapid 5 mL washes in ice-cold binding buffer.
Bound radioactivity is measured by gamma emission spectrometry.
Reference Example 4
Serotonin Transporter (SERT) Membrane Radioligand Binding
Assays
[0315] Total cell membranes are prepared from HEK-293 cells
expressing the recombinant human serotonin transporter (hSERT)
grown to confluence in 150 mm tissue culture dishes. Cells are
scraped into standard medium and pelleted at 1600 g. The medium is
discarded and the pellet resuspended in 5 mL per plate of ice-cold
binding buffer (100 mM NaCl, 50 mM Tris, pH 7.4 at room
temperature) by trituration, and the cells are repelleted at 20,000
g. Supernatant is discarded and cells are resuspended in binding
buffer (50 mM Tris-HCl, pH 7.4, 120 mM NaCl, 5 mM KCl) and
homogenized with a polytron (Brinkman) at 25,000 revs/min for 5 s.
Centrifugation, resuspension, and homogenization are repeated, and
a sample of suspension is used for Bradford protein determination
(BioRad). Samples of membrane suspensions are frozen at -80.degree.
C. prior to use. Typical yields are about 100 .mu.g membrane
protein per 106 cells. Assays performed in duplicate are initiated
with 0.4 nM [3H]paroxetine. Non-specific binding is determined by
the inclusion of 10 .mu.M imipramine. Incubation is carried out for
60 min at 25.degree. C. Assays are terminated by rapid filtration
over GF/B glass-fiber filters soaked in 0.5% polyethylineimine
using an automated cell harvester (Brandel) followed by three rapid
5 mL washes in ice-cold binding buffer. Bound radioactivity is
measured by beta emission spectrometry.
Reference Example 5
Pharmacological Activity for Glaucoma Assay
[0316] Pharmacological activity for glaucoma treatment can be
demonstrated using assays designed to test the ability of the
subject compounds to decrease intraocular pressure. Examples of
such assays are described in the following reference, incorporated
herein by reference: C. Liljebris, G. Selen, B. Resul, J.
Sternschantz, and U. Hacksell, "Derivatives of
17-phenyl-18,19,20-trinorprostaglandin F.sub.2 Ispropyl Ester:
Potential Anti-glaucoma Agents", Journal of Medicinal Chemistry
1995, 38 (2): 289-304.
Reference Example 6
Functional Screen for NET Activity Modulators
[0317] At the center of the cellular assay used to characterized
inhibition of the human norepinephrine transporter (hNET) is a
fluorophore which mimics a biogenic amine and is actively
transported in to the cell. After incubation with a potential
inhibitor, the dye solution is added in the presence of a
fluorescence masking dye. When the fluorescent dye is transported
into the cell and removed from the masking dye, light is emitted at
a wavelength of 510 nm when excited with light of 425 nm.
Inhibitors of the norepinephrine transporter prevent this
time-dependent increase in fluorescence. (Blakely R D, DeFelice L
J, and Galli A. A. Physiol. 2005. 20, 225-231).
[0318] HEK-293 cells recombinantly over-expressing the human
Norepinephrine Transporter are grown in phenol red-free DMEM medium
(Gibco #21063-029) containing 10% fetal bovine serum (Atlanta
Biologicals #S11050), 100 units/mL penicillin/streptomycin (Gibco
#15140-122), and 250 .mu.g/mL G-418 (Gibco #10131027) at 37.degree.
C. and 5% CO.sub.2.
[0319] One day prior to testing, cells are plated into black, clear
bottom plates (Costar #3904) at a density of 100,000 per well. On
the morning of the assay, culture media is removed and replaced
with 100 .mu.L of HBSS (1.times. Hank's Balanced Salt Solution
containing 20 mM HEPES) (10.times.HBSS Gibco #14065, 1 M HEPES
Gibco #15630-80) containing test compound. Cells are exposed to the
test compound (or DMSO in control wells) for 30 min. At the end of
the pre-incubation period, 10 .mu.L of 10.times. dye solution is
added and thoroughly mixed with the compound solution. After a 30
min incubation period, fluorescence is quantified using an Analyst
HT fluorescence plate reader (Molecular Devices, Sunnyvale,
Calif.).
[0320] While the invention has been described in detail and with
reference to specific embodiments thereof, it will be apparent to
one skilled in the art that various changes and modifications can
be made without departing from the spirit and scope of the
invention.
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