U.S. patent application number 12/215367 was filed with the patent office on 2009-02-12 for modulators of muscarinic receptors.
Invention is credited to Miguel Garcia-Guzman Blanco, Peter D. J. Grootenhuis, Philip Martin Londo, Lewis R. Makings.
Application Number | 20090042928 12/215367 |
Document ID | / |
Family ID | 34676813 |
Filed Date | 2009-02-12 |
United States Patent
Application |
20090042928 |
Kind Code |
A1 |
Grootenhuis; Peter D. J. ;
et al. |
February 12, 2009 |
Modulators of muscarinic receptors
Abstract
The modulator compounds described herein modulate muscarinic
receptors and are useful for treating muscarinic receptor mediated
diseases.
Inventors: |
Grootenhuis; Peter D. J.;
(San Diego, CA) ; Blanco; Miguel Garcia-Guzman;
(San Diego, CA) ; Makings; Lewis R.; (Encinitas,
CA) ; Londo; Philip Martin; (Santee, CA) |
Correspondence
Address: |
MILLER CANFIELD PADDOCK AND STONE PLC
277 SOUTH ROSE STREET, SUITE 5000
KALAMAZOO
MI
49007
US
|
Family ID: |
34676813 |
Appl. No.: |
12/215367 |
Filed: |
June 26, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11005944 |
Dec 7, 2004 |
7446112 |
|
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12215367 |
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60528049 |
Dec 9, 2003 |
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Current U.S.
Class: |
514/300 ;
546/122 |
Current CPC
Class: |
C07D 471/04 20130101;
A61P 25/04 20180101; A61P 25/16 20180101; A61P 25/18 20180101; A61P
11/06 20180101; A61P 25/00 20180101; A61P 25/28 20180101; A61P
17/02 20180101; A61P 1/02 20180101; A61P 9/04 20180101; A61P 25/24
20180101; A61P 25/14 20180101; A61P 27/04 20180101; A61P 3/04
20180101; A61P 1/04 20180101; A61P 27/02 20180101 |
Class at
Publication: |
514/300 ;
546/122 |
International
Class: |
A61K 31/4375 20060101
A61K031/4375; C07D 471/04 20060101 C07D471/04; A61P 25/00 20060101
A61P025/00 |
Claims
1. A compound of Formula I, ##STR00086## wherein: Ring A is aryl or
heteroaryl; One of V, W, Y or Z is nitrogen and the other of V, W,
Y and Z are --C(R.sup.5)--; Each R.sup.1 is independently selected
from H, aliphatic, cycloaliphatic, heteroaliphatic and heterocycle,
wherein each of the aliphatic, cycloaliphatic, heteroaliphatic and
heterocycle is optionally substituted with 1-3 Ra; Each R.sup.2 is
independently selected from H, aryl, heteroaryl, aliphatic,
cycloaliphatic, heteroaliphatic, heterocycle, --C(O)Rc, and
--S(O).sub.2Rc, wherein each aliphatic, cycloaliphatic,
heteroaliphatic and heterocycle is optionally substituted with 1-3
Ra, and wherein each aryl and heteroaryl is optionally substituted
with 1-3 Rb, or R.sup.1 and R.sup.2 together with the nitrogen to
which they are attached may form a heterocyclic ring or a
heteroaryl ring each optionally substituted with 1-3 Ra; Each
R.sup.3 is independently H, halo, haloaliphatic, aliphatic, --ORd,
or --S(O).sub.iRd; Each R.sup.4 is independently selected from H,
halogen, --CN, --OH, --NO.sub.2, --ORd, --C(O)Rd, --C(O)ORd,
--C(O)N(Rd).sub.2, --N(Rd).sub.2, --N(Rd)C(O)Rd, --N(Rd)C(O)ORd,
--OC(O)ORd, --OC(O)NRd, --N(Rd)S(O).sub.2Rd, aliphatic optionally
substituted with 1-3 Ra, and any two adjacent R.sup.4 on Ring A
together with the atoms to which they are attached may be taken
together to form a heterocyclic or carbocyclic ring; Each R.sup.5
is independently selected from H, halogen, --CN, --OH, --NO.sub.2,
--ORd, --C(O)Rd, --C(O)ORd, --C(O)N(Rd).sub.2, --N(Rd).sub.2,
--N(Rd)C(O)Rd, --N(Rd)C(O)ORd, --OC(O)ORd, --OC(O)NRd,
--N(Rd)S(O).sub.2Rd, aliphatic optionally substituted with 1-3 of
Ra, and any two adjacent R.sup.5 together with the atoms to which
they are attached may be taken together to form a heterocyclic or
carbocyclic ring; Each Ra is independently selected from aryl,
heteroaryl, halogen, --CN, --OH, --ORd, --C(O)Rd, --C(O)ORd,
--C(O)N(Rd).sub.2, --N(Rd).sub.2, --NRdC(O)Rd, --N(Rd)C(O)ORd,
--N(Rd)C(O)N(Rd).sub.2, --OC(O)ORd, --OC(O)N(Rd).sub.2, .dbd.N--OH,
.dbd.NORd, .dbd.N.dbd.N(Rd).sub.2, .dbd.O, .dbd.S,
--S(O).sub.2N(Rd).sub.2, --N(Rd)S(O).sub.2Rd,
--N(Rd)S(O).sub.2N(Rd).sub.2 and --S(O).sub.iRd; Each Rb is
independently selected from halo, aryl, --OH, --ORd,
--S(O).sub.iRd, --N(Rd).sub.2, --NRdC(O)Rd, --NRdC(O)ORd, --C(O)Rd,
--C(O)ORd, --C(O)N(Rd).sub.2, --S(O).sub.iN(Rd).sub.2, --CN, and
--NO.sub.2; Each Rc is independently selected from H, aliphatic,
cycloaliphatic, heteroaliphatic, heterocycle, aryl, heteroaryl,
--ORd, and --N(Rd).sub.2, wherein the aliphatic, cycloaliphatic,
heteroaliphatic, heterocycle, aryl, and heteroaryl are optionally
substituted with 1-3 of Ra; Each Rd is independently selected from
H, aliphatic, heteroaliphatic, heterocycle, cycloaliphatic, aryl,
heteroaryl, wherein each of aliphatic, heteroaliphatic,
heterocycle, cycloaliphatic, aryl, heteroaryl may be optionally
substituted with 1-3 of halo, aryl, --OH, --Oaliphatic, --Oaryl,
--Oacyl, --NH.sub.2, --N(aliphatic).sub.2, --N(aryl).sub.2,
--S(O).sub.ialiphatic, or --S(O).sub.iaryl; n is 0 to 3; and i is 0
to 2; and provided (i) when A is 2-trifluoromethylphenyl, one of Z
or Y is N and the remaining of W, V, Y, or Z is --C(H)--, R.sup.3
is H, and R.sup.1 is H, then R.sup.2 is not indazolyl, pyrazolyl,
or triazolyl each optionally substituted with aliphatic, phenyl,
and --C(O)O-aliphatic; (ii) when A is phenyl, V is N and W, Y and Z
are --C(H)--, R.sup.3 is H, that R.sup.1 and R.sup.2 together with
the nitrogen to which they are bound do not form piperidine
optionally substituted with --C(O)O-aliphatic,
--C(O)N(H)-aliphatic, or --C(O)OH; (iii) when A is phenyl, V is N
and W, Y and Z are --C(H)--, R.sup.3 is H, and R.sup.1 is H, that
R.sup.2 is not --CH.sub.2-piperidine; (iv) when A is phenyl
optionally substituted with one R.sup.4, Z is N, V and W are
--C(H)--, and R.sup.3 is H, that Y is other than --C(aliphatic)-;
(v) when A is phenyl optionally substituted with one R.sup.4, Z is
N, Y and V are --C(H)--, and R.sup.3 is H, that W is other than
--C(aliphatic)-; and (vi) when A is pyridinyl, Z is N, R.sup.1 is
H, R.sup.3 is H, and V, W, and Y are --C(H)--, that R.sup.2 is not
--(CH.sub.2).sub.2--N(aliphatic).sub.2.
2. The compound of claim 1, wherein one of W, Y, Z and V is N and
the other of W,Y,Z and V is C--R.
3. The compound of claim 2, wherein W is N and Y, Z and V are
C--R.sup.5.
4. The compound of claim 3, wherein one of R.sup.1 and R.sup.2 is
H.
5. The compound of claim 4, wherein both of R.sup.1 and R.sup.2 are
H.
6. The compound of claim 4, wherein A is aryl.
7. The compound of claim 5, wherein R.sup.4 is selected from the
group consisting of halogen, --ORd, and aliphatic optionally
substituted with 1-3 of Ra.
8. The compound of claim 7, wherein R.sup.4 is --Oaliphatic or
haloaliphatic.
9. A compound selected from the group consisting of
2-Phenyl-[1,6]naphthyridin-4-ylamine,
2-(3-Methoxy-phenyl)-[1,6]naphthyridin-4-ylamine,
2-Benzo[1,3]dioxol-5-yl-[1,6]naphthyridin-4-ylamine,
2-(3-Chloro-phenyl)-[1,6]naphthyridin-4-ylamine,
2-m-Tolyl-[1,6]naphthyridin-4-ylamine,
2-(3-Trifluoromethyl-phenyl)-[1,6]naphthyridin-4-ylamine,
2-(4-Fluoro-phenyl)-[1,6]naphthyridin-4-ylamine,
2-(4-Chloro-phenyl)-[1,6]naphthyridin-4-ylamine,
2-(4-Methoxy-phenyl)-[1,6]naphthyridin-4-ylamine,
2-p-Tolyl-[1,6]naphthyridin-4-ylamine,
2-(4-Trifluoromethyl-phenyl)-[1,6]naphthyridin-4-ylamine,
2-(3-Fluoro-phenyl)-[1,6]naphthyridin-4-ylamine, and
2-(2,3-Dihydro-benzo[1,4]dioxin-6-yl)-[1,6]naphthyridin-4-ylamine.
10. The compound of claim 2, wherein Y is N and W, Z and V are
C--R.sup.5.
11. The compound of claim 10, wherein R.sup.1, R.sup.2 and R.sup.5
are H.
12. The compound of claim 9, wherein A is aryl.
13. The compound of claim 13, wherein R.sup.4 is selected from the
group consisting of halogen, --ORd, and aliphatic optionally
substituted with 1-3 of Ra.
14. The compound of claim 13, wherein R.sup.4 is --Oaliphatic or
haloaliphatic.
15. A compound selected from the group consisting of
2-Phenyl-[1,7]naphthyridin-4-ylamine,
2-(3-Chloro-phenyl)-[1,7]naphthyridin-4-ylamine,
2-(3-Methoxy-phenyl)-[1,7]naphthyridin-4-ylamine, and
2-Benzo[1,3]dioxol-5-yl-[1,7]naphthyridin-4-ylamine.
16. The compound of claim 2, wherein Z is N and Y,W and V are
C--R.sup.5.
17. The compound of claim 16, wherein R.sup.1 and R.sup.2 are
H.
18. The compound of claim 17, wherein A is aryl.
19. The compound of claim 16, wherein R.sup.4 is selected from the
group consisting of halogen, --ORd, and aliphatic optionally
substituted with 1-3 of Ra.
20. The compound of claim 19, wherein R.sup.4 is --Oaliphatic or
haloaliphatic.
21. A compound selected from the group consisting of
2-Phenyl-[1,8]naphthyridin-4-ylamine,
2-(3-Methoxy-phenyl)-[1,8]naphthyridin-4-ylamine,
2-Benzo[1,3]dioxol-5-yl-[1,8]naphthyridin-4-ylamine, and
2-(3-Chloro-phenyl)-[1,8]naphthyridin-4-ylamine.
22. The compound of claim 1, wherein A is heteroaryl.
23. A compound selected from the group comprising
2-Pyridin-2-yl-[1,6]naphthyridin-4-ylamine,
2-Pyridin-2-yl-[1,7]naphthyridin-4-ylamine, and
2-Pyridin-2-yl-[1,8]naphthyridin-4-ylamine.
24. A method of modulating activity of a muscarinic receptor,
comprising the step of contacting said receptor with a compound of
formula I: ##STR00087## wherein: Ring A is aryl or heteroaryl; One
of V, W, Y or Z is nitrogen and the other of V, W, Y and Z are
--C(R.sup.5)--; Each R.sup.1 is independently selected from H,
aliphatic, cycloaliphatic, heteroaliphatic and heterocycle, wherein
each of the aliphatic, cycloaliphatic, heteroaliphatic and
heterocycle is optionally substituted with 1-3 Ra; Each R.sup.2 is
independently selected from H, aryl, heteroaryl, aliphatic,
cycloaliphatic, heteroaliphatic, heterocycle, --C(O)Rc, and
--S(O).sub.2Rc, wherein each aliphatic, cycloaliphatic,
heteroaliphatic and heterocycle is optionally substituted with 1-3
Ra, and wherein each aryl and heteroaryl is optionally substituted
with 1-3 Rb, or R.sup.1 and R.sup.2 together with the nitrogen to
which they are attached may form a heterocyclic ring or a
heteroaryl ring each optionally substituted with 1-3 Ra; Each
R.sup.3 is independently H, halo, haloaliphatic, --CN, aliphatic,
--ORd, or --S(O).sub.iRd; Each R.sup.4 is independently selected
from H, halogen, --CN, --OH, --NO.sub.2, --ORd, --C(O)Rd,
--C(O)ORd, --C(O)N(Rd).sub.2, --N(Rd).sub.2, --N(Rd)C(O)Rd,
--N(Rd)C(O)ORd, --OC(O)ORd, --OC(O)NRd, --N(Rd)S(O).sub.2Rd,
aliphatic optionally substituted with 1-3 Ra, and any two adjacent
R.sup.4 on Ring A together with the atoms to which they are
attached may be taken together to form a heterocyclic or
carbocyclic ring; Each R.sup.5 is independently selected from H,
halogen, --CN, --OH, --NO.sub.2, --ORd, --C(O)Rd, --C(O)ORd,
--C(O)N(Rd).sub.2, --N(Rd).sub.2, --N(Rd)C(O)Rd, --N(Rd)C(O)ORd,
--OC(O)ORd, --OC(O)NRd, --N(Rd)S(O).sub.2Rd, aliphatic optionally
substituted with 1-3 of Ra, and any two adjacent R.sup.5 together
with the atoms to which they are attached may be taken together to
form a heterocyclic or carbocyclic ring; Each Ra is independently
selected from aryl, heteroaryl, halogen, --CN, --OH, --ORd,
--C(O)Rd, --C(O)ORd, --C(O)N(Rd).sub.2, --N(Rd).sub.2, --NRdC(O)Rd,
--N(Rd)C(O)ORd, --N(Rd)C(O)N(Rd).sub.2, --OC(O)ORd,
--OC(O)N(Rd).sub.2, .dbd.N--OH, .dbd.NORd, .dbd.N.dbd.N(Rd).sub.2,
.dbd.O, .dbd.S, --S(O).sub.2N(Rd).sub.2, --N(Rd)S(O).sub.2Rd,
--N(Rd)S(O).sub.2N(Rd).sub.2 and --S(O).sub.iRd; Each Rb is
independently selected from halo, aryl, --OH, --ORd,
--S(O).sub.iRd, --N(Rd).sub.2, --NRdC(O)Rd, --NRdC(O)ORd, --C(O)Rd,
--C(O)ORd, --C(O)N(Rd).sub.2, --S(O).sub.iN(Rd).sub.2, --CN, and
--NO.sub.2; Each Rc is independently selected from H, aliphatic,
cycloaliphatic, heteroaliphatic, heterocycle, aryl, heteroaryl,
--ORd, and --N(Rd).sub.2, wherein the aliphatic, cycloaliphatic,
heteroaliphatic, heterocycle, aryl, and heteroaryl are optionally
substituted with 1-3 of Ra; Each Rd is independently selected from
H, aliphatic, heteroaliphatic, heterocycle, cycloaliphatic, aryl,
heteroaryl, wherein each of aliphatic, heteroaliphatic,
heterocycle, cycloaliphatic, aryl, heteroaryl may be optionally
substituted with 1-3 of halo, aryl, --OH, --Oaliphatic, --Oaryl,
--Oacyl, --NH.sub.2, --N(aliphatic).sub.2, --N(aryl).sub.2,
--S(O).sub.ialiphatic, or --S(O).sub.iaryl; n is 0 to 3; and i is 0
to 2.
25. The method of claim 24, wherein W is N and Y, Z and V are
C--R.sup.5.
26. The method of claim 24, wherein one of R.sup.1 and R.sup.2 is
H.
27. The method of claim 26, wherein both of R.sup.1 and R.sup.2 are
H.
28. The method of claim 24, wherein R.sup.4 is selected from the
group consisting of halogen, --ORd, and aliphatic optionally
substituted with 1-3 of Ra.
29. The method of claim 28, wherein R.sup.4 is --Oaliphatic or
haloaliphatic.
30. The method of claim 24, wherein Ring A is aryl optionally
substituted with 1-4 of R.sup.4.
31. The method of claim 24, wherein Ring A is heteroaryl optionally
substituted with 1-4 of R.sup.4.
32. The method of claim 24, wherein the compound is selected from
the group consisting of 2-Phenyl-[1,6]naphthyridin-4-ylamine,
2-(3-Methoxy-phenyl)-[1,6]naphthyridin-4-ylamine,
2-Benzo[1,3]dioxol-5-yl-[1,6]naphthyridin-4-ylamine,
2-(3-Chloro-phenyl)-[1,6]naphthyridin-4-ylamine,
2-m-Tolyl-[1,6]naphthyridin-4-ylamine,
2-(3-Trifluoromethyl-phenyl)-[1,6]naphthyridin-4-ylamine,
2-(4-Fluoro-phenyl)-[1,6]naphthyridin-4-ylamine,
2-(4-Chloro-phenyl)-[1,6]naphthyridin-4-ylamine,
2-(4-Methoxy-phenyl)-[1,6]naphthyridin-4-ylamine,
2-p-Tolyl-[1,6]naphthyridin-4-ylamine,
2-(4-Trifluoromethyl-phenyl)-[1,6]naphthyridin-4-ylamine,
2-(3-Fluoro-phenyl)-[1,6]naphthyridin-4-ylamine,
2-(2,3-Dihydro-benzo[1,4]dioxin-6-yl)-[1,6]naphthyridin-4-ylamine.
2-Phenyl-[1,7]naphthyridin-4-ylamine,
2-(3-Chloro-phenyl)-[1,7]naphthyridin-4-ylamine,
2-(3-Methoxy-phenyl)-[1,7]naphthyridin-4-ylamine,
2-Benzo[1,3]dioxol-5-yl-[1,7]naphthyridin-4-ylamine,
2-Phenyl-[1,8]naphthyridin-4-ylamine,
2-(3-Methoxy-phenyl)-[1,8]naphthyridin-4-ylamine,
2-Benzo[1,3]dioxol-5-yl-[1,8]naphthyridin-4-ylamine,
2-(3-Chloro-phenyl)-[1,8]naphthyridin-4-ylamine.
2-Pyridin-2-yl-[1,6]naphthyridin-4-ylamine,
2-Pyridin-2-yl-[1,7]naphthyridin-4-ylamine, and
2-Pyridin-2-yl-[1,8]naphthyridin-4-ylamine.
33. A pharmaceutical composition comprising a compound a compound
according to claim 1 and a pharmaceutical carrier.
34. A method of treating or reducing the severity of a muscarinic
receptor mediated disease in a mammal, comprising the step of
administering to said mammal a compound as described in claim 1 or
a composition according to claim 33.
35. The method of claim 34, wherein said receptor is M.sub.1.
36. A method of treating or reducing the severity of a disease in a
patient, wherein said disease is selected from CNS derived
pathologies including cognitive disorders, Attention Deficit
Hyperactivity Disorder (ADHD), obesity, Alzheimer's disease,
various dementias such as vascular dementia, psychosis associated
with CNS disorders including schizophrenia, mania, bipolar
disorders, pain conditions including acute and chronic syndromes,
Huntington's Chorea, Friederich's ataxia, Gilles de la Tourette's
Syndrome, Downs Syndrome, Pick disease, clinical depression,
Parkinson's disease, peripheral disorders such as reduction of
intra ocular pressure in Glaucoma and treatment of dry eyes and dry
mouth including Sjogren's Syndrome, bradhycardia, gastric acid
secretion, asthma, GI disturbances, and wound healing, wherein said
method comprises the step of contacting said patient with a
compound as described in claim 1 or a composition according to
claim 33.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This patent application is a divisional application of U.S.
patent application Ser. No. 11/005,944, which was filed on Dec. 7,
2004 and claims priority to U.S. Provisional Application Ser. No.
60/528,049, which was filed on Dec. 9, 2003. Both of these
underlying applications are incorporated herein by reference.
TECHNICAL FIELD OF THE INVENTION
[0002] The present invention relates to compounds useful as
therapeutic agents. The invention also provides pharmaceutically
acceptable compositions comprising the compounds of the invention
and methods of using the compositions in the treatment of various
disorders.
BACKGROUND OF THE INVENTION
[0003] The neurotransmitter acetylcholine binds to two types of
cholinergic receptors: the ionotropic family of nicotinic receptors
and the metabotropic family of muscarinic receptors. Muscarinic
receptors belong to the large superfamily of plasma membrane-bound
G protein coupled receptors (GPCRs). To date, five subtypes of
muscarinic receptors (M.sub.1-M.sub.5) have been cloned and
sequenced from a variety of species, and show a remarkably high
degree of homology across species and receptor subtype. These
M.sub.1-M.sub.5 muscarinic receptors are predominantly expressed
within the parasympathetic nervous system which exerts excitatory
and inhibitory control over the central and peripheral tissues and
participate in a number of physiologic functions, including heart
rate, arousal, cognition, sensory processing, and motor
control.
[0004] Muscarinic agonists such as muscarine and pilocarpine, and
antagonists, such as atropine have been known for over a century,
but little progress has been made in the discovery of receptor
subtype-selective compounds, thereby making it difficult to assign
specific functions to the individual receptors. See, e.g., DeLapp,
N. et al., "Therapeutic Opportunities for Muscarinic Receptors in
the Central Nervous System," J. Med. Chem., 43(23), pp. 4333-4353
(2000); Hulme, E. C. et al., "Muscarinic Receptor Subtypes," Ann.
Rev. Pharmacol. Toxicol., 30, pp. 633-673 (1990); Caulfield, M. P.
et al., "Muscarinic Receptors-Characterization, Coupling, and
Function," Pharmacol. Ther., 58, pp. 319-379 (1993); Caulfield, M.
P. et al., International Union of Pharmacology. XVII.
Classification of Muscarinic Acetylcholine Receptors," Pharmacol.
Rev., 50, pp. 279-290 (1998), the disclosures of which are
incorporated herein by reference.
[0005] The Muscarinic family of receptors is the target of a large
number of pharmacological agents used for various diseases,
including leading drugs for COPD, asthma, urinary incontinence,
glaucoma, Alzheimer's (AchE inhibitors), and Pain.
[0006] Pain can be roughly divided into three different types:
acute, inflammatory, and neuropathic. Acute pain serves an
important protective function in keeping the organism safe from
stimuli that may produce tissue damage. Severe thermal, mechanical,
or chemical inputs have the potential to cause severe damage to the
organism if unheeded. Acute pain serves to quickly remove the
individual from the damaging environment. Acute pain by its very
nature generally is short lasting and intense. Inflammatory pain on
the other had may last for much longer periods of time and it's
intensity is more graded. Inflammation may occur for many reasons
including tissue damage, autoimmune response, and pathogen
invasion. Inflammatory pain is mediated by an "inflammatory soup"
that consists of substance P, histamines, acid, prostaglandin,
bradykinin, CGRP, cytokines, ATP, and neurotransmitter release. The
third class of pain is neuropathic and involves nerve damage that
results in reorganization of neuronal proteins and circuits
yielding a pathologic "sensitized" state that can produce chronic
pain lasting for years. This type of pain provides no adaptive
benefit and is particularly difficult to treat with existing
therapies.
[0007] Pain, particularly neuropathic and intractable pain is a
large unmet medical need. Millions of individuals suffer from
severe pain that is not well controlled by current therapeutics.
The current drugs used to treat pain include NSAIDS, COX2
inhibitors, opioids, tricyclic antidepressants, and
anticonvulsants. Neuropathic pain has been particularly difficult
to treat as it does not respond well to opiods until high doses are
reached. Gabapentin is currently the favored therapeutic for the
treatment of neuropathic pain although it works in only 60% of
patients where it shows modest efficacy. The drug is however very
safe and side effects are generally tolerable although sedation is
an issue at higher doses.
[0008] Despite the large therapeutic value of this family,
cholinergic drugs are limited by the lack of selectivity of these
agents, with significant activation of the parasympathetic
autonomous system and elevated incidence of adverse effects. The
molecular cloning of the muscarinic receptors and the
identification of the physiological role of specific isoforms using
knock-out mice, has recently delineated novel opportunities for
selective muscarinic ligands, and has helped to define the
selectivity profile that is required for enhanced efficacy and
reduced side effects.
[0009] There is a need for modulators of muscarinic receptors
M.sub.1-M.sub.5. There is also a need for methods for treating
muscarinic receptor-mediated diseases.
[0010] There is also a need for modulators of muscarinic receptors
that are selective as to subtypes M.sub.1-M.sub.5.
SUMMARY OF THE INVENTION
[0011] The present invention provides a method of modulating
activity of a muscarinic receptor, comprising the step of
contacting said receptor with a compound of formula I:
##STR00001##
wherein:
[0012] Ring A is aryl or heteroaryl;
[0013] One of V, W, Y or Z is nitrogen and the other of V, W, Y and
Z are --C(R.sup.5)--;
[0014] Each R.sup.1 is independently selected from H, aliphatic,
cycloaliphatic, heteroaliphatic and heterocycle, wherein each of
the aliphatic, cycloaliphatic, heteroaliphatic and heterocycle is
optionally substituted with 1-3 Ra;
[0015] Each R.sup.2 is independently selected from H, aryl,
heteroaryl, aliphatic, cycloaliphatic, heteroaliphatic,
heterocycle, --C(O)Rc, and --S(O).sub.2Rc, wherein each aliphatic,
cycloaliphatic, heteroaliphatic and heterocycle is optionally
substituted with 1-3 Ra, and wherein each aryl and heteroaryl is
optionally substituted with 1-3 Rb, or [0016] R.sup.1 and R.sup.2
together with the nitrogen to which they are attached may form a
heterocyclic ring or a heteroaryl ring each optionally substituted
with 1-3 Ra;
[0017] Each R.sup.3 is independently H, halo, haloaliphatic,
aliphatic, --ORd, or --S(O).sub.iRd; Each R.sup.4 is independently
selected from H, halogen, --CN, --OH, --NO.sub.2, --ORd, --C(O)Rd,
--C(O)ORd, --C(O)N(Rd).sub.2, --N(Rd).sub.2, --N(Rd)C(O)Rd,
--N(Rd)C(O)ORd, --OC(O)ORd, --OC(O)NRd, --N(Rd)S(O).sub.2Rd,
aliphatic optionally substituted with 1-3 Ra, and any two adjacent
R.sup.4 on Ring A together with the atoms to which they are
attached may be taken together to form a heterocyclic or
carbocyclic ring;
[0018] Each R.sup.5 is independently selected from H, halogen,
--CN, --OH, --NO.sub.2, --ORd, --C(O)Rd, --C(O)ORd,
--C(O)N(Rd).sub.2, --N(Rd).sub.2, --N(Rd)C(O)Rd, --N(Rd)C(O)ORd,
--OC(O)ORd, --OC(O)NRd, --N(Rd)S(O).sub.2Rd, aliphatic optionally
substituted with 1-3 of Ra, and any two adjacent R.sup.5 together
with the atoms to which they are attached may be taken together to
form a heterocyclic or carbocyclic ring;
[0019] Each Ra is independently selected from aryl, heteroaryl,
halogen, --CN, --OH, --ORd, --C(O)Rd, --C(O)ORd, --C(O)N(Rd).sub.2,
--N(Rd).sub.2, --NRdC(O)Rd, --N(Rd)C(O)ORd, --N(Rd)C(O)N(Rd).sub.2,
--OC(O)ORd, --OC(O)N(Rd).sub.2, .dbd.N--OH, .dbd.NORd,
.dbd.N.dbd.N(Rd).sub.2, .dbd.O, .dbd.S, --S(O).sub.2N(Rd).sub.2,
--N(Rd)S(O).sub.2Rd, --N(Rd)S(O).sub.2N(Rd).sub.2 and
--S(O).sub.iRd;
[0020] Each Rb is independently selected from halo, aryl, --OH,
--ORd, --S(O).sub.iRd, --N(Rd).sub.2, --NRdC(O)Rd, --NRdC(O)ORd,
--C(O)Rd, --C(O)ORd, --C(O)N(Rd).sub.2, --S(O).sub.iN(Rd).sub.2,
--CN, and --NO.sub.2;
[0021] Each Rc is independently selected from H, aliphatic,
cycloaliphatic, heteroaliphatic, heterocycle, aryl, heteroaryl,
--ORd, and --N(Rd).sub.2, wherein the aliphatic, cycloaliphatic,
heteroaliphatic, heterocycle, aryl, and heteroaryl are optionally
substituted with 1-3 of Ra;
[0022] Each Rd is independently selected from H, aliphatic,
heteroaliphatic, heterocycle, cycloaliphatic, aryl, and heteroaryl,
wherein each of aliphatic, heteroaliphatic, heterocycle,
cycloaliphatic, aryl, heteroaryl may be optionally substituted with
1-3 of halo, aryl, --OH, --Oaliphatic, --Oaryl, --Oacyl,
--NH.sub.2, --N(aliphatic).sub.2, --N(aryl).sub.2,
--S(O).sub.ialiphatic, or --S(O).sub.iaryl;
[0023] n is 0 to 3; and
[0024] i is 0 to 2.
[0025] The present invention also provides compounds of formula
(I), compositions comprising compounds of formula (I), and methods
of treating muscarinic receptor mediated diseases using compounds
of formula (I).
[0026] Advantageously, the compounds of the invention unexpectedly
modulate muscarinic receptors.
DETAILED DESCRIPTION OF THE INVENTION
I. Definitions
[0027] As used herein, the following definitions shall apply unless
otherwise indicated. For purposes of this invention, the chemical
elements are identified in accordance with the Periodic Table of
the Elements, CAS version, Handbook of Chemistry and Physics, 75th
Ed. Additionally, general principles of organic chemistry are
described in "Organic Chemistry", Thomas Sorrell, University
Science Books, Sausalito: 1999, and "March's Advanced Organic
Chemistry", 5th Ed., Ed.: Smith, M. B. and March, J., John Wiley
& Sons, New York: 2001, the entire contents of which are hereby
incorporated by reference.
[0028] The term "muscarinic receptor," without a prefix specifying
the receptor subtype, refers to one or more of the five receptor
subtypes M.sub.1-M.sub.5.
[0029] The term "modulating" as used herein means increasing or
decreasing, e.g. activity, by a measurable amount. Compounds that
modulate muscarinic activity by increasing the activity of the
muscarinic receptors are called agonists. Compounds that modulate
muscarinic activity by decreasing the activity of the muscarinic
receptors are called antagonists. An agonist interacts with a
muscarinic receptor to increase the ability of the receptor to
transduce an intracellular signal in response to endogenous ligand
binding. An antagonist interacts with a muscarinic receptor and
competes with the endogenous ligand(s) or substrate(s) for binding
site(s) on the receptor to decrease the ability of the receptor to
transduce an intracellular signal in response to endogenous ligand
binding.
[0030] The phrase "treating or reducing the severity of a
muscarinic receptor mediated disease" refers both to treatments for
diseases that are directly caused by muscarinic activities and
alleviation of symptoms of diseases not directly caused by
muscarinic activities. Examples of diseases whose symptoms may be
affected by muscarinic activity include, but are not limited to,
CNS derived pathologies including cognitive disorders, Attention
Deficit Hyperactivity Disorder (ADHD), obesity, Alzheimer's
disease, various dementias such as vascular dementia, psychosis
including schizophrenia, mania, bipolar disorders, pain conditions
including acute and chronic syndromes, Huntington's Chorea,
Friederich's ataxia, Gilles de la Tourette's Syndrome, Downs
Syndrome, Pick disease, clinical depression, Parkinson's disease,
peripheral disorders such as reduction of intra ocular pressure in
Glaucoma and treatment of dry eyes and dry mouth including
Sjogren's Syndrome, bradhycardia, gastric acid secretion, asthma,
GI disturbances and wound healing.
[0031] The phrase "optionally substituted" is used interchangeably
with the phrase "substituted or unsubstituted." As described
herein, compounds of the invention may optionally be substituted
with one or more substituents, such as are illustrated generally
above, or as exemplified by particular classes, subclasses, and
species of the invention. It will be appreciated that the phrase
"optionally substituted" is used interchangeably with the phrase
"substituted or unsubstituted." In general, the term "substituted",
whether preceded by the term "optionally" or not, refers to the
replacement of hydrogen radicals in a given structure with the
radical of a specified substituent. Unless otherwise indicated, an
optionally substituted group may have a substituent at each
substitutable position of the group, and when more than one
position in any given structure may be substituted with more than
one substituent selected from a specified group, the substituent
may be either the same or different at every position. As one of
ordinary skill in the art will recognize, combinations of
substituents envisioned by this invention are those combinations
that result in the formation of stable or chemically feasible
compounds.
[0032] The phrase "stable or chemically feasible", as used herein,
refers to compounds that are not substantially altered when
subjected to conditions to allow for their production, detection,
and preferably their recovery, purification, and use for one or
more of the purposes disclosed herein. In some embodiments, a
stable compound or chemically feasible compound is one that is not
substantially altered when kept at a temperature of 40.degree. C.
or less, in the absence of moisture or other chemically reactive
conditions, for at least a week.
[0033] The term "aliphatic" or "aliphatic group", as used herein,
means a straight-chain (i.e., unbranched) or branched, substituted
or unsubstituted hydrocarbon chain that is completely saturated or
that contains one or more units of unsaturation, or a monocyclic
hydrocarbon or bicyclic hydrocarbon that is completely saturated or
that contains one or more units of unsaturation, but which is not
aromatic (also referred to herein as "carbocycle" "cycloaliphatic"
or "cycloaliphatic"), that has a single point of attachment to the
rest of the molecule. Unless otherwise specified, aliphatic groups
contain 1-20 aliphatic carbon atoms. In some embodiments, aliphatic
groups contain 1-10 aliphatic carbon atoms. In other embodiments,
aliphatic groups contain 1-8 aliphatic carbon atoms. In still other
embodiments, aliphatic groups contain 1-6 aliphatic carbon atoms,
and in yet other embodiments aliphatic groups contain 1-4 aliphatic
carbon atoms. In some embodiments, "cycloaliphatic" (or
"carbocycle") refers to a monocyclic C.sub.3-C.sub.8 hydrocarbon or
bicyclic C.sub.8-C.sub.12 hydrocarbon that is completely saturated
or that contains one or more units of unsaturation, but which is
not aromatic, that has a single point of attachment to the rest of
the molecule wherein any individual ring in said bicyclic ring
system has 3-7 members. Suitable aliphatic groups include, but are
not limited to, linear or branched, substituted or unsubstituted
aliphatic, alkenyl, alkynyl groups and hybrids thereof such as
(cycloaliphatic)aliphatic, (cycloalkenyl)aliphatic or
(cycloaliphatic)alkenyl.
[0034] The term "heteroaliphatic", as used herein, means aliphatic
groups wherein one or two carbon atoms are independently replaced
by one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon.
Heteroaliphatic groups may be substituted or unsubstituted,
branched or unbranched, cyclic or acyclic, and include
"heterocycle", "heterocyclyl", "heterocycloaliphatic", or
"heterocyclic" groups.
[0035] The term "heterocycle", "heterocyclyl",
"heterocycloaliphatic", or "heterocyclic" as used herein means
non-aromatic, monocyclic, bicyclic, or tricyclic ring systems in
which one or more ring members are an independently selected
heteroatom. In some embodiments, the "heterocycle", "heterocyclyl",
"heterocycloaliphatic", or "heterocyclic" group has three to
fourteen ring members in which one or more ring members is a
heteroatom independently selected from oxygen, sulfur, nitrogen, or
phosphorus, and each ring in the system contains 3 to 7 ring
members.
[0036] The term "heteroatom" means one or more of oxygen, sulfur,
nitrogen, phosphorus, or silicon (including, any oxidized form of
nitrogen, sulfur, phosphorus, or silicon; the quaternized form of
any basic nitrogen or; a substitutable nitrogen of a heterocyclic
ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in
pyrrolidinyl) or NR.sup.+ (as in N-substituted pyrrolidinyl)).
[0037] The term "unsaturated", as used herein, means that a moiety
has one or more units of unsaturation.
[0038] The term "alkoxy", or "thioaliphatic", as used herein,
refers to an aliphatic group, as previously defined, attached to
the principal carbon chain through an oxygen ("alkoxy") or sulfur
("thioaliphatic") atom.
[0039] The terms "haloaliphatic", "haloalkenyl" and "haloalkoxy"
means aliphatic, alkenyl or alkoxy, as the case may be, substituted
with one or more halogen atoms. The term "halogen" means F, Cl, Br,
or I.
[0040] The term "aryl" used alone or as part of a larger moiety as
in "araliphatic", "aralkoxy", or "aryloxyaliphatic", refers to
monocyclic, bicyclic, and tricyclic ring systems having a total of
five to fourteen ring members, wherein at least one ring in the
system is aromatic and wherein each ring in the system contains 3
to 7 ring members. The term "aryl" may be used interchangeably with
the term "aryl ring".
[0041] The term "heteroaryl", used alone or as part of a larger
moiety as in "heteroaraliphatic" or "heteroarylalkoxy", refers to
monocyclic, bicyclic, and tricyclic ring systems having a total of
five to fourteen ring members, wherein at least one ring in the
system is aromatic, at least one ring in the system contains one or
more heteroatoms, and wherein each ring in the system contains 3 to
7 ring members. The term "heteroaryl" may be used interchangeably
with the term "heteroaryl ring" or the term "heteroaromatic".
[0042] The term "amino protecting group" refers to a suitable
chemical group that may be attached to a nitrogen atom. The term
"protected" refers to when the designated functional group is
attached to a suitable chemical group (protecting group). Examples
of suitable amino protecting groups and protecting groups are
described in T. W. Greene and P. G. M. Wuts, Protective Groups in
Organic Synthesis, 2d. Ed., John Wiley and Sons (1991); L. Fieser
and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis,
John Wiley and Sons (1994); L. Paquette, ed. Encyclopedia of
Reagents for Organic Synthesis, John Wiley and Sons (1995) and are
exemplified in certain of the specific compounds used in this
invention.
[0043] Unless otherwise stated, structures depicted herein are also
meant to include all isomeric (e.g., enantiomeric, diastereomeric,
and geometric (or conformational)) forms of the structure; for
example, the R and S configurations for each asymmetric center, (Z)
and (E) double bond isomers, and (Z) and (E) conformational
isomers. Therefore, single stereochemical isomers as well as
enantiomeric, diastereomeric, and geometric (or conformational)
mixtures of the present compounds are within the scope of the
invention. Unless otherwise stated, all tautomeric forms of the
compounds of the invention are within the scope of the invention.
Additionally, unless otherwise stated, structures depicted herein
are also meant to include compounds that differ only in the
presence of one or more isotopically enriched atoms. For example,
compounds having the present structures except for the replacement
of hydrogen by deuterium or tritium, or the replacement of a carbon
by a .sup.13C-- or .sup.14C-enriched carbon are within the scope of
this invention. Such compounds are useful, for example, as
analytical tools or probes in biological assays.
II. Description of Compounds
[0044] In general, the compounds useful for modulating muscarinic
activity of muscarinic receptors have the formula I:
##STR00002##
Wherein Ring A, R.sup.1, R.sup.2, R.sup.3, R.sup.4, V, W, Y, Z, and
n are defined above.
[0045] In some aspects, Ring A in formula I is an aryl optionally
substituted with 1-3 of R.sup.4. Embodiments of this aspect include
one or more of the following features. Ring A is phenyl. Ring A is
substituted with 1 or 2 of R.sup.4. R.sup.4 is aliphatic. R.sup.4
is CH.sub.3. R.sup.4 is halogen. R.sup.4 is --ORd. R.sup.4 is
--Oaliphatic. R.sup.4 is --CH.sub.3. R.sup.4 is haloaliphatic. Two
R.sup.4 substituents on Ring A together form --O--CH.sub.2--O--.
Two R.sup.4 substituents on Ring A together form
--O--CH.sub.2--CH.sub.2--O--. Z is N. Y is N. W is N. V is N. n is
0. n is 1. n is 2.
[0046] In other aspects, A in formula I is an heteroaryl optionally
substituted with 1-3 of R.sup.4. Embodiments of this aspect include
one or more of the following features. Ring A is pyridinyl. Ring A
is substituted with 1 or 2 of R.sup.4. R.sup.4 is aliphatic.
R.sup.4 is CH.sub.3. R.sup.4 is halogen. R.sup.4 is --ORd. R.sup.4
is --Oaliphatic. R.sup.4 is --OCH.sub.3. R.sup.4 is haloaliphatic.
Two R.sup.4 substituents on Ring A together form
--O--CH.sub.2--O--. Two R.sup.4 substituents on Ring A together
form --O--CH.sub.2--CH.sub.2--O--. Z is N. Y is N. W is N. V is N.
n is 0. n is 1. n is 2.
[0047] In other embodiments, the compounds of formula I include one
or more of the following features.
[0048] R.sup.3 is H, halo, aliphatic, or --ORd. R.sup.3 is H.
[0049] R.sup.1 is H, aliphatic, or heteroaliphatic, wherein each of
the aliphatic and heteroaliphatic is optionally substituted with
1-3 Ra. Each R.sup.1 is independently selected from cycloaliphatic
and heterocycle each optionally substituted with 1-3 Ra. R.sup.1 is
H. R.sup.1 is aliphatic optionally substituted with 1-3 Ra.
[0050] R.sup.2 is H, aliphatic, heteroaliphatic, --C(O)Rc, or
--S(O).sub.2Rc, wherein each aliphatic and heteroaliphatic is
optionally substituted with 1-3 Ra. R.sup.2 is H. R.sup.2 is
--C(O)Rc. R.sup.2 is aliphatic optionally substituted with 1-3 Ra.
Each R.sup.2 is independently selected from aryl and heteroaryl,
each optionally substituted with 1-3 Ra, and wherein each aryl and
heteroaryl is optionally substituted with 1-3 Rb. Each R.sup.2 is
independently selected from cycloaliphatic and heterocycle each
optionally substituted with 1-3 Ra.
[0051] R.sup.1 and R.sup.2 together with the nitrogen to which they
are attached may form a heterocyclic ring optionally substituted
with 1-3 Ra.
[0052] Each R.sup.5 is independently selected from H, halogen,
--ORd, --C(O)Rd, --C(O)ORd, --C(O)N(Rd).sub.2, and aliphatic
optionally substituted with 1-3 of Ra. Each R.sup.5 is
independently selected from H, --N(Rd).sub.2, --N(Rd)C(O)Rd,
--N(Rd)C(O)ORd, --OC(O)ORd, --OC(O)NRd, and
--N(Rd)S(O).sub.2Rd.
[0053] Each Ra is independently selected from halogen, --CN, --ORd,
--C(O)Rd, --C(O)ORd, --C(O)N(Rd).sub.2, --N(Rd).sub.2, --NRdC(O)Rd,
--N(Rd)C(O)ORd, --N(Rd)C(O)N(Rd).sub.2, --OC(O)ORd,
--OC(O)N(Rd).sub.2, .dbd.O, --S(O).sub.2N(Rd).sub.2,
--N(Rd)S(O).sub.2Rd, --N(Rd)S(O).sub.2N(Rd).sub.2 and
--S(O).sub.iRd. Each Ra is independently selected from halogen,
--ORd, --C(O)Rd, --C(O)ORd, --C(O)N(Rd).sub.2, --N(Rd).sub.2,
--OC(O)ORd, --OC(O)N(Rd).sub.2, and .dbd.O. Ra is independently
selected from halogen, --CN, --ORd, --C(O)Rd, --C(O)ORd, and
.dbd.O.
[0054] Each Rb is independently selected from halo, aryl, --ORd,
--S(O).sub.iRd, --N(Rd).sub.2, --NRdC(O)Rd, --NRdC(O)ORd, --C(O)Rd,
--C(O)ORd, --C(O)N(Rd).sub.2, --S(O).sub.iN(Rd).sub.2, --CN, and
--NO.sub.2. Each Rb is independently selected from halo, aryl,
--ORd, --C(O)Rd, --C(O)ORd, --C(O)N(Rd).sub.2, and --CN. Each Rb is
independently selected from halo, aryl, --S(O).sub.iRd,
--N(Rd).sub.2, --NRdC(O)Rd, --NRdC(O)ORd, --S(O).sub.iN(Rd).sub.2,
and --NO.sub.2. Each Rb is independently selected from halo, --ORd,
--S(O).sub.iRd, and --S(O).sub.iN(Rd).sub.2. Each Rb is
independently selected from halo and --ORd, --C(O)Rd, and
--C(O)ORd.
[0055] Each Rc is independently selected from H, aliphatic,
heteroaliphatic, --ORd, and --N(Rd).sub.2, wherein the aliphatic
and heteroaliphatic are optionally substituted with 1-3 of Ra. Each
Rc is independently selected from H, --ORd, and --N(Rd).sub.2. Each
Rc is independently selected from aliphatic and heteroaliphatic
each of which are optionally substituted with 1-3 of Ra. Each Rc is
independently aliphatic optionally substituted with 1-3 of Ra.
[0056] Each Rd is independently selected from H, aliphatic,
heteroaliphatic, wherein each of the aliphatic and heteroaliphatic
are optionally substituted with 1-3 of halo, --OH, --Oaliphatic,
--Oaryl, --Oacyl, --NH.sub.2, --N(aliphatic).sub.2,
--N(aryl).sub.2, --S(O).sub.ialiphatic, or --S(O).sub.iaryl. Each
Rd is independently selected from H, heterocycle, and
cycloaliphatic, wherein each of the heterocycle and
cycloaliphaticare optionally substituted with 1-3 of halo, aryl,
--OH, --Oaliphatic, --Oaryl, --Oacyl, --NH.sub.2,
--N(aliphatic).sub.2, --N(aryl).sub.2, --S(O).sub.ialiphatic, or
--S(O).sub.iaryl. Each Rd is independently selected from H, aryl,
and heteroaryl, wherein each of the aryl and heteroaryl are
optionally substituted with 1-3 of halo, aryl, --OH, --Oaliphatic,
--Oaryl, --Oacyl, --NH.sub.2, --N(aliphatic).sub.2,
--N(aryl).sub.2, --S(O).sub.ialiphatic, or --S(O).sub.iaryl. Each
Rd is independently selected from H and aliphatic optionally
substituted with 1-3 of halo, aryl, --OH, --Oaliphatic, --Oaryl,
--Oacyl, --NH.sub.2, --N(aliphatic).sub.2, --N(aryl).sub.2,
--S(O).sub.ialiphatic, or --S(O).sub.iaryl.
[0057] In other embodiments, the compounds have the structure of
formula I provided
[0058] (i) when A is 2-trifluoromethylphenyl, one of Z or Y is N
and the remaining of W, V, Y, or Z is --C(H)--, R.sup.3 is H, and
R.sup.1 is H, then R.sup.2 is not indazolyl, pyrazolyl, or
triazolyl each optionally substituted with aliphatic, phenyl, and
--C(O)O-aliphatic;
[0059] (ii) when A is phenyl, V is N and W, Y and Z are --C(H)--,
R.sup.3 is H, that R.sup.1 and R.sup.2 together with the nitrogen
to which they are bound do not form piperidine optionally
substituted with --C(O)O-aliphatic, --C(O)N(H)-aliphatic, or
--C(O)OH;
[0060] (iii) when A is phenyl, V is N and W, Y and Z are --C(H)--,
R.sup.3 is H, and R.sup.1 is H, that R.sup.2 is not
--CH.sub.2-piperidine;
[0061] (iv) when A is phenyl optionally substituted with one
R.sup.4, Z is N, V and W are --C(H)--, and R.sup.3 is H, that Y is
other than --C(aliphatic)-;
[0062] (v) when A is phenyl optionally substituted with one
R.sup.4, Z is N, Y and V are --C(H)--, and R.sup.3 is H, that W is
other than --C(aliphatic)-;
[0063] (vi) when A is pyridinyl, Z is N, R.sup.1 is H, R.sup.3 is
H, and V, W, and Y are --C(H)--, that R.sup.2 is not
--(CH.sub.2).sub.2--N(aliphatic).sub.2.
[0064] In still other aspects, the invention features compounds of
formula I that include combinations of the different aspects and
embodiments described above. For instance, embodiments of compounds
of formula I where Ring A is aryl may include one or more of the
embodiments described above for R.sup.1, R.sup.2, R.sup.3, R.sup.5,
Ra, Rb, Rc, and Rd.
[0065] Exemplary compound of formula I are show in Table 1.
TABLE-US-00001 TABLE 1 Exemplary Compounds of Formula I. 1
##STR00003## 2 ##STR00004## 3 ##STR00005## 4 ##STR00006## 5
##STR00007## 6 ##STR00008## 7 ##STR00009## 8 ##STR00010## 9
##STR00011## 10 ##STR00012## 11 ##STR00013## 12 ##STR00014## 13
##STR00015## 14 ##STR00016## 15 ##STR00017## 16 ##STR00018## 17
##STR00019## 18 ##STR00020## 19 ##STR00021## 20 ##STR00022## 21
##STR00023## 22 ##STR00024## 23 ##STR00025## 24 ##STR00026## 25
##STR00027## 26 ##STR00028## 27 ##STR00029##
III. General Synthetic Methodology
[0066] The compounds of this invention may be prepared in general
by methods known to those skilled in the art for analogous
compounds, as illustrated by the general schemes below, and the
preparative examples that follow. Starting materials are
commercially available from typical chemical reagent supply
companies, such as, Aldrich Chemicals Co., Sigma Chemical Company,
and the like. Compounds that are not commercially available can be
prepared by those of ordinary skill in art following procedures set
forth in references such as, "Fieser and Fieser's Reagents for
Organic Synthesis", Volumes 1-15, John Wiley and Sons, 1991;
"Rodd's Chemistry of Carbon Compounds", Volumes 1-5 and
Supplementals, Elservier Science Publishers, 1989; and "Organic
Reactions", Volumes 1-40, John Wiley and Sons, 1991.
##STR00030##
[0067] Directed metalation of optionally substituted pyridines
followed by quenching by a suitable reagent, such as ethyl oxalate,
afforded the corresponding .alpha.-ketoesters. Removal of the
protecting group followed by condensation with a suitable ketone,
for example an acetophenone, and acid-catalyzed cyclization
afforded various [l,n]naphthyridine carboxylates in moderate to
good yields. These compounds were subjected to the Curtius
rearrangement to afford the corresponding amino naphthyridines. The
preparation of certain compounds of the present invention are
taught in Examples 1 to 4.
[0068] Other methods for producing the compounds are known in the
art. For example, 1,5-naphthyridines may be prepared according to
methods described in U.S. Application Nos. 20030212084 and
20040152704, European Application No. 487242, and PCT Publication
Nos. WO9943682 and WO0047580. Each of these references is
incorporated herein.
[0069] In still further aspects, the compound of the formula II are
useful in preparing the compound of formula I
##STR00031##
wherein each of Ring A, V, W, Y, Z, R.sup.3, R.sup.4 and n are
defined above.
IV. Uses, Formulations, Compositions, and Administration
[0070] The present invention includes within its scope
pharmaceutically acceptable prodrugs of the compounds of the
present invention. A "pharmaceutically acceptable prodrug" means
any pharmaceutically acceptable salt, ester, salt of an ester, or
other derivative of a compound of the present invention which, upon
administration to a recipient, is capable of providing (directly or
indirectly) a compound of this invention or an active metabolite or
residue thereof. Preferred prodrugs are those that increase the
bioavailability of the compounds of this invention when such
compounds are administered to a mammal or which enhance delivery of
the parent compound to a biological compartment relative to the
parent species.
[0071] The term "pharmaceutically acceptable carrier, adjuvant, or
vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that
does not destroy the pharmacological activity of the compound with
which it is formulated. Pharmaceutically acceptable carriers,
adjuvants or vehicles that may be used in the compositions of this
invention include, but are not limited to, ion exchangers, alumina,
aluminum stearate, lecithin, serum proteins, such as human serum
albumin, buffer substances such as phosphates, glycine, sorbic
acid, potassium sorbate, partial glyceride mixtures of saturated
vegetable fatty acids, water, salts or electrolytes, such as
protamine sulfate, disodium hydrogen phosphate, potassium hydrogen
phosphate, sodium chloride, zinc salts, colloidal silica, magnesium
trisilicate, polyvinyl pyrrolidone, cellulose-based substances,
polyethylene glycol, sodium carboxymethylcellulose, polyacrylates,
waxes, polyethylene-polyoxypropylene-block polymers, polyethylene
glycol and wool fat.
[0072] Pharmaceutically acceptable salts of the compounds of this
invention include those derived from pharmaceutically acceptable
inorganic and organic acids and bases. Examples of suitable acid
salts include acetate, adipate, alginate, aspartate, benzoate,
benzenesulfonate, bisulfate, butyrate, citrate, camphorate,
camphorsulfonate, cyclopentanepropionate, digluconate,
dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptanoate, glycerophosphate, glycolate, hemisulfate,
heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide,
2-hydroxyethanesulfonate, lactate, maleate, malonate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,
oxalate, palmoate, pectinate, persulfate, 3-phenylpropionate,
phosphate, picrate, pivalate, propionate, salicylate, succinate,
sulfate, tartrate, thiocyanate, tosylate and undecanoate. Other
acids, such as oxalic, while not in themselves pharmaceutically
acceptable, may be employed in the preparation of salts useful as
intermediates in obtaining the compounds of the invention and their
pharmaceutically acceptable acid addition salts.
[0073] Salts derived from appropriate bases include alkali metal
(e.g., sodium and potassium), alkaline earth metal (e.g., calcium
or magnesium), ammonium and N.sup.+ (C.sub.1-4 alkyl).sub.4 salts
or salts of lysine and arginine. This invention also envisions the
quaternization of any basic nitrogen-containing groups of the
compounds disclosed herein. Water or oil-soluble or dispersible
products may be obtained by such quaternization. Other salts can be
found in "Practical Process, Research, & Development,"
Anderson, Neal G., Academic Press, 2000, the contents of which are
incorporated herein by reference.
[0074] The compositions of the present invention may be
administered orally, parenterally, by inhalation spray, topically,
rectally, intermuscularly, subcutaneously, nasally, buccally,
vaginally or via an implanted reservoir. The term "parenteral" as
used herein includes subcutaneous, intravenous, intramuscular,
intra-articular, intra-synovial, intrasternal, intrathecal,
intrahepatic, intralesional and intracranial injection or infusion
techniques. Preferably, the compositions are administered orally,
intraperitoneally or intravenously. Sterile injectable forms of the
compositions of this invention may be aqueous or oleaginous
suspension. These suspensions may be formulated according to
techniques known in the art using suitable dispersing or wetting
agents and suspending agents. The sterile injectable preparation
may also be a sterile injectable solution or suspension in a
non-toxic parenterally-acceptable diluent or solvent, for example
as a solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution and
isotonic sodium chloride solution. In addition, sterile, fixed oils
are conventionally employed as a solvent or suspending medium.
[0075] For this purpose, any bland fixed oil may be employed
including synthetic mono- or di-glycerides. Fatty acids, such as
oleic acid and its glyceride derivatives are useful in the
preparation of injectables, as are natural
pharmaceutically-acceptable oils, such as olive oil or castor oil,
especially in their polyoxyethylated versions. These oil solutions
or suspensions may also contain a long-chain alcohol diluent or
dispersant, such as carboxymethyl cellulose or similar dispersing
agents that are commonly used in the formulation of
pharmaceutically acceptable dosage forms including emulsions and
suspensions. Other commonly used surfactants, such as Tweens, Spans
and other emulsifying agents or bioavailability enhancers which are
commonly used in the manufacture of pharmaceutically acceptable
solid, liquid, or other dosage forms may also be used for the
purposes of formulation.
[0076] The pharmaceutically acceptable compositions of this
invention may be orally administered in any orally acceptable
dosage form including, but not limited to, capsules, tablets,
aqueous suspensions or solutions. In the case of tablets for oral
use, carriers commonly used include lactose and corn starch.
Lubricating agents, such as magnesium stearate, are also typically
added. For oral administration in a capsule form, useful diluents
include lactose and dried cornstarch. When aqueous suspensions are
required for oral use, the active ingredient is combined with
emulsifying and suspending agents. If desired, certain sweetening,
flavoring or coloring agents may also be added.
[0077] Alternatively, the pharmaceutically acceptable compositions
of this invention may be administered in the form of suppositories
for rectal administration. These can be prepared by mixing the
agent with a suitable non-irritating excipient that is solid at
room temperature but liquid at rectal temperature and therefore
will melt in the rectum to release the drug. Such materials include
cocoa butter, beeswax and polyethylene glycols.
[0078] The pharmaceutically acceptable compositions of this
invention may also be administered topically, especially when the
target of treatment includes areas or organs readily accessible by
topical application, including diseases of the eye, the skin, or
the lower intestinal tract. Suitable topical formulations are
readily prepared for each of these areas or organs.
[0079] Topical application for the lower intestinal tract can be
effected in a rectal suppository formulation (see above) or in a
suitable enema formulation. Topically-transdermal patches may also
be used.
[0080] For topical applications, the pharmaceutically acceptable
compositions may be formulated in a suitable ointment containing
the active component suspended or dissolved in one or more
carriers. Carriers for topical administration of the compounds of
this invention include, but are not limited to, mineral oil, liquid
petrolatum, white petrolatum, propylene glycol, polyoxyethylene,
polyoxypropylene compound, emulsifying wax and water.
Alternatively, the pharmaceutically acceptable compositions can be
formulated in a suitable lotion or cream containing the active
components suspended or dissolved in one or more pharmaceutically
acceptable carriers. Suitable carriers include, but are not limited
to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl
esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and
water.
[0081] For ophthalmic use, the pharmaceutically acceptable
compositions may be formulated as micronized suspensions in
isotonic, pH adjusted sterile saline, or, preferably, as solutions
in isotonic, pH adjusted sterile saline, either with or without a
preservative such as benzylalkonium chloride. Alternatively, for
ophthalmic uses, the pharmaceutically acceptable compositions may
be formulated in an ointment such as petrolatum.
[0082] The pharmaceutically acceptable compositions of this
invention may also be administered by nasal aerosol or inhalation.
Such compositions are prepared according to techniques well-known
in the art of pharmaceutical formulation and may be prepared as
solutions in saline, employing benzyl alcohol or other suitable
preservatives, absorption promoters to enhance bioavailability,
fluorocarbons, and/or other conventional solubilizing or dispersing
agents.
[0083] Most preferably, the pharmaceutically acceptable
compositions of this invention are formulated for oral
administration.
[0084] The amount of the compounds of the present invention that
may be combined with the carrier materials to produce a composition
in a single dosage form will vary depending upon the host treated,
the particular mode of administration. Preferably, the compositions
should be formulated so that a dosage of between 0.01-100 mg/kg
body weight/day of the modulator can be administered to a patient
receiving these compositions.
[0085] It should also be understood that a specific dosage and
treatment regimen for any particular patient will depend upon a
variety of factors, including the activity of the specific compound
employed, the age, body weight, general health, sex, diet, time of
administration, rate of excretion, drug combination, and the
judgment of the treating physician and the severity of the
particular disease being treated. The amount of a compound of the
present invention in the composition will also depend upon the
particular compound in the composition.
[0086] Depending upon the particular condition, or disease, to be
treated or prevented, additional therapeutic agents, which are
normally administered to treat or prevent that condition, may also
be present in the compositions of this invention. As used herein,
additional therapeutic agents that are normally administered to
treat or prevent a particular disease, or condition, are known as
"appropriate for the disease, or condition, being treated."
[0087] According to a preferred embodiment, the compounds of
formula I are selective modulators of M.sub.1, M.sub.2 and M.sub.4.
More preferably, the compounds of formula (I, IA, II, and III) are
selective modulators of M.sub.1 and M.sub.4. Or, the compounds of
formula (I, IA, II, and III) are selective modulators of M.sub.2
and M.sub.4. Yet more preferably, the compounds of formula (I, IA,
II, and III) are selective modulators of one of M.sub.1, M.sub.2,
and M.sub.4. The compounds of formula (I, IA, II, and III) are
selective modulators of M.sub.4. The compounds of formula (I, IA,
II, and III) are selective modulators of M.sub.1.
[0088] Applicants believe that the ability of the compounds of the
present invention to modulate the activity of muscarinic receptors
is derived from the affinity of these compounds to the muscarinic
receptors. Such affinity, applicants believe, activates a
muscarinic receptor (i.e, an agonist) or inhibits the activity of a
muscarinic receptor.
[0089] According to another embodiment, the compounds of formula
(I, IA, II, and III) are selective activators of all of M.sub.1,
M.sub.2, and M.sub.4. In other embodiments, the compounds of
formula (I, IA, II, and III) are selective activators of one of
M.sub.1, M.sub.2, and M.sub.4 and selective inhibitors of the other
two of M.sub.1, M.sub.2, and M.sub.4. In another embodiment, the
compounds of formula (I, IA, II, and III) are selective activators
of up to two of M.sub.1, M.sub.2, and M.sub.4 and selective
inhibitors of the other of M.sub.1, M.sub.2, and M.sub.4. In still
another embodiment, the compounds of formula (I, IA, II, and III)
are selective inhibitors of all of M.sub.1, M.sub.2, and
M.sub.4.
[0090] According to another embodiment, the compounds of compounds
of formula (I, IA, II, and III) are selective inhibitors of one or
more of M.sub.1, M.sub.2, or M.sub.4. In one embodiment,
preferably, the compounds of formula (I, IA, II, and III) are
selective inhibitors of M.sub.4. In another embodiment, the
compounds of formula I are selective inhibitors of M.sub.1. In yet
another embodiment, the compounds of formula I are selective
inhibitors of M.sub.1 and M.sub.4. In still another embodiment, the
compounds of formula I are selective inhibitors of M.sub.1 and
M.sub.2 or M.sub.4 and M.sub.2.
[0091] The term "selective" as used herein means a measurably
greater ability to modulate one muscarinic receptor subtype when
compared to the other muscarinic receptor subtypes. E.g., the term
"selective M.sub.4 agonist" means a compound that has a measurably
greater ability to act as an M.sub.4 agonist when compared to that
compound's agonist activity with the other muscarinic receptor
subtype(s).
[0092] According to an alternative embodiment, the present
invention provides a method of treating a muscarinic receptor
mediated disease in a mammal, comprising the step of administering
to said mammal a composition comprising a compound of formula I, or
a preferred embodiment thereof as set forth above.
[0093] According to a preferred embodiment, the present invention
provides a method of treating a disease mediated by one or more of
M.sub.1, M.sub.2, or M.sub.4, comprising the step of administering
to said mammal a composition comprising a compound of formula (I,
IA, II, and III), or a preferred embodiment thereof as set forth
above. Or in another embodiment the disease is mediated by M.sub.2.
Or, said disease is mediated by M.sub.1. Yet more preferably, said
disease is mediated by M.sub.4. In still further embodiments, the
disease is mediate by all of M.sub.1, M.sub.2, and M.sub.4. In
another embodiment, the disease is mediate by two of M.sub.1,
M.sub.2, and M.sub.4.
[0094] According to a preferred embodiment, the present invention
provides a method of treating or reducing the severity of a disease
in a patient, wherein said disease is selected from CNS derived
pathologies including cognitive disorders, Attention Deficit
Hyperactivity Disorder (ADHD), obesity, Alzheimer's disease,
various dementias such as vascular dementia, psychosis associated
with CNS disorders including schizophrenia, mania, bipolar
disorders, pain conditions including acute and chronic syndromes,
Huntington's Chorea, Friederich's ataxia, Gilles de la Tourette's
Syndrome, Downs Syndrome, Pick disease, clinical depression,
Parkinson's disease, peripheral disorders such as reduction of
intra ocular pressure in Glaucoma and treatment of dry eyes and dry
mouth including Sjogren's Syndrome, and wound healing, wherein said
method comprises the step of contacting said patient with a
compound according to the present invention.
[0095] In one embodiment, the present invention provides a method
for the treatment or lessening the severity of acute, chronic,
neuropathic, or inflammatory pain, arthritis, migrane, cluster
headaches, trigeminal neuralgia, herpetic neuralgia, general
neuralgias, epilepsy or epilepsy conditions, neurodegenerative
disorders, psychiatric disorders such as anxiety and depression,
myotonia, arrythmia, movement disorders, neuroendocrine disorders,
ataxia, multiple sclerosis, irritable bowel syndrome, incontinence,
visceral pain, osteoarthritis pain, postherpetic neuralgia,
diabetic neuropathy, radicular pain, sciatica, back pain, head or
neck pain, severe or intractable pain, nociceptive pain,
breakthrough pain, postsurgical pain, or cancer pain is provided
comprising administering an effective amount of a compound, or a
pharmaceutically acceptable composition comprising a compound to a
subject in need thereof. In certain embodiments, a method for the
treatment or lessening the severity of acute, chronic, neuropathic,
or inflammatory pain is provided comprising administering an
effective amount of a compound or a pharmaceutically acceptable
composition to a subject in need thereof. In certain other
embodiments, a method for the treatment or lessening the severity
of radicular pain, sciatica, back pain, head pain, or neck pain is
provided comprising administering an effective amount of a compound
or a pharmaceutically acceptable composition to a subject in need
thereof. In still other embodiments, a method for the treatment or
lessening the severity of severe or intractable pain, acute pain,
post-surgical pain, back pain, or cancer pain is provided
comprising administering an effective amount of a compound or a
pharmaceutically acceptable composition to a subject in need
thereof.
[0096] According to an alternative embodiment, the present
invention provides a method of treating or reducing the severity of
a disease in a patient, wherein said disease is selected from pain,
psychosis (including schizophrenia, hallucinations, and delusions),
Alzheimer's disease, Parkinson's disease, glaucoma, bradhycardia,
gastric acid secretion, asthma, GI disturbances or wound
healing.
[0097] According to a preferred embodiment, the present invention
is useful for treating or reducing the severity of psychosis,
Alzheimer's disease, pain, or Parkinson's disease.
[0098] In order that the invention described herein may be more
fully understood, the following examples are set forth. It should
be understood that these examples are for illustrative purposes
only and are not to be construed as limiting this invention in any
manner.
[0099] All references cited above are incorporated herein by
reference.
[0100] Other embodiments of the compounds of formula I are shown
below. The following examples are illustrative of the compounds of
formula I and are not meant to be limiting.
EXAMPLES
Example 1
Preparation of 2,2-Dimethyl-N-pyridinyl-propionamides
##STR00032##
[0101] 1A: 2,2-Dimethyl-N-pyridin-4-yl-propionamide
[0102] Following procedures generally taught in J. A. Turner, J.
Org. Chem. 1983, 48, 3401-3408, a solution of pivaloyl chloride
(13.5 mL, 110 mmol) in CH.sub.2Cl.sub.2 (20 mL) was slowly added to
an ice-cold solution of 4-aminopyridine (9.41 g, 100 mmol) and
triethylamine (17.4 mL, 125 mmol) in CH.sub.2Cl.sub.2 (150 mL).
After addition was complete, the resulting mixture was warmed to
room temperature and stirred for 2 h. The solution was poured into
water, the CH.sub.2Cl.sub.2 layer was washed with dilute
NaHCO.sub.3, dried over Na2SO4, and evaporated to leave a light
brown solid. Recrystallization from EtOAc/hexanes afforded the
product as a white crystal, which was collected by vacuum
filtration (13.03 g, 73%). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.50 (d, J=4.8 Hz, 2H), 7.50 (d, J=4.8 Hz, 2H), 7.44 (br s,
1H), 1.33 (s, 9H). MS (LR-APCI) calcd. for C.sub.10H.sub.15N.sub.2O
(M+H) 179.1; found 179.1.
1B: 2,2-Dimethyl-N-pyridin-2-yl-propionamide
##STR00033##
[0104] Following the procedure taught in Example 1A, the title
compound was prepared having the following characteristics (white
powder, 14.4 g. 81%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
8.28-8.24 (m, 2H), 7.99 (br s, 1H), 7.72-7.68 (m, 1H), 7.05-7.02
(m, 1H), 1.71 (s, 9H). MS (LR-APCI) calcd. for
C.sub.10H.sub.15N.sub.2O (M+H) 179.1; found 179.1.
1C: 2,2-Dimethyl-N-pyridin-3-yl-propionamide
##STR00034##
[0106] Following the procedure taught in Example IA, the title
compound was prepared having the following characteristics (white
solid, 13.90 g, 78%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
8.56-8.54 (m, 1H), 8.36-8.34 (m, 1H), 8.22-8.18 (m, 1H), 7.39 (br
s, 1H), 7.30-7.26 (m, 1H), 1.34 (s, 9H). MS (LR-APCI) calcd. for
C.sub.10H.sub.15N.sub.2O (M+H) 179.1; found 179.2.
Example 2
Preparation of (2,2-Dimethyl-propionylamino)-pyridinyl -oxo-acetic
acid ethyl esters
2A: [4-(2,2-Dimethyl-propionylamino)-pyridin-3-yl]-oxo-acetic acid
ethyl ester
##STR00035##
[0108] Following procedures generally taught in J. A. Turner, J.
Org. Chem. 1983, 48, 3401-8; and C. Rivalle & E. Bisagni, J.
Heterocyclic Chem., 1997, 34, 441-4, a 3-necked, 500 mL round
bottom flask with thermometer and addition funnel were flame-dried
under N.sub.2. 2,2-Dimethyl-N-pyridin-3-yl-propionamide (4.46 g,
25.0 mmol) was added, followed by THF (50 mL). The solution was
cooled to -78.degree. C. (to control the pending exotherm), and
n-BuLi (39 mL of a 1.6 M solution in hexanes, 2.5 equiv) was added
dropwise via addition funnel with vigorous stirring of the slurry,
keeping internal temp below -50.degree. C. during addition. Once
the n-BuLi addition was complete (the solution was yellow and
homogeneous), the mixture was warmed to 0.degree. C. for 3 h (a
white precipitate emerges). The solution was cooled back down to
-78.degree. C. and diethyl oxalate (8.84 mL, 2.6 equiv) in THF (13
mL) was added dropwise via syringe (mild exotherm observed). Once
the addition was complete, the reaction was stirred 15 min at
-78.degree. C., then warmed to room temperature over 15 min and
stirred an additional 15 min (a dark red-orange solution emerges as
the solution was stirred at room temperature). The mixture was
poured onto ice and extracted with Et.sub.2O, washing the extract
once with water. The solution was dried (MgSO.sub.4), filtered, and
concentrated to a dark orange oil. Purification by biotage (50%
EtOAc/hexanes) afforded the product as an orange oil (3.61 g, 52%).
R.sub.f (prod)=0.57 (50% EtOAc/hexanes). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 10.69 (s, 1H), 8.79 (s, 1H), 8.66 (d, J=6.4
Hz, 1H), 8.00 (d, J=5.6 Hz, 1H), 4.32 (q, J=7.0 Hz, 2H), 1.28 (t,
J=7.0 Hz, 3H), 1.21 (s, 9H). MS (LR-APCI) calcd. for
C.sub.14H.sub.19N.sub.2O.sub.4 (M+H) 279.1; found 279.1.
2B: [2-(2,2-Dimethyl-propionylamino)-pyridin-3-yl]-oxo-acetic acid
ethyl ester
##STR00036##
[0110] Following the procedure taught in Example 2A, the title
compound was prepared having the following characteristics
(off-white solid, 1.90 g, 27%). R.sub.f (prod)=0.36 (50%
EtOAc/hexanes). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.48
(s, 1H), 8.56-8.53 (m, 1H), 7.99-7.95 (m, 1H), 7.35-7.30 (m, 1H),
4.21-4.14 (m, 2H), 1.25-1.19 (m, 3H), 1.14 (s, 9H). (t, J=7.0 Hz,
3H), 1.21 (s, 9H). MS (LR-APCI) calcd. for
C.sub.14H.sub.19N.sub.2O.sub.4 (M+H) 279.1; found 279.0.
2C: [3-(2,2-Dimethyl-propionylamino)-pyridin-4-yl]-oxo-acetic acid
ethyl ester
##STR00037##
[0112] Following the procedure taught in Example 2A, the title
compound was prepared having the following characteristics (yellow
solid, 1.57 g, 23%). R.sub.f (prod)=0.22 (40% EtOAc/hexanes).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.83 (s, 1H), 10.15 (s,
1H), 8.52 (d, J=5.6 Hz, 1H), 7.57 (d, J=5.6 Hz, 1H), 4.50 (q, J=7.0
Hz, 2H), 1.45 (t, J=7.0 Hz, 3H), 1.37 (s, 9H). MS (LR-APCI) calcd.
for C.sub.14H.sub.19N.sub.2O.sub.4 (M+H) 279.1; found 279.1.
Example 3
Preparation of Naphthyridine-4-carboxylic acids
3A: 2-Phenyl-[1,6]naphthyridine-4-carboxylic acid
##STR00038##
[0114] [4-(2,2-Dimethyl-propionylamino)-pyridin-3-yl]-oxo-acetic
acid ethyl ester (532 mg, 1.91 mmol) was taken up in ethanol (1.9
mL) and 4N KOH (1.9 mL, 4 equiv) was added. The solution was
refluxed for 2-3 h at 100.degree. C., acetophenone (0.446 mL, 2.0
equiv) was added neat, and the reaction was allowed to reflux
overnight. Upon cooling to room temperature, the solution was
diluted with 1N NaOH and extracted twice with ether. The product
was precipitated from the aqueous layer by the addition of glacial
AcOH (5-10 mL). The solids were collected by vacuum filtration,
rinsed with cold water, and dried under high vacuum (white solid,
371 mg, 78%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.00 (s,
1H), 8.80 (d, J=5.6 Hz, 1H), 8.56 (s, 1H), 8.35-8.31 (m, 2H), 8.03
(d, J=6.0 Hz, 1H), 7.61-7.58 (m, 3H). MS (LR-APCI) calcd. for
C.sub.15H.sub.11N.sub.2O.sub.2 (M+H) 251.1; found 251.5.
3B: 2-Phenyl-[1,8]naphthyridine-4-carboxylic acid
##STR00039##
[0116] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics
(off-white solid, 380 mg, 84%). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.16-9.14 (m, 1H), 9.12-9.09 (m, 1H), 8.57 (s, 1H),
8.35-8.32 (m, 2H), 7.71 (dd, J=8.6, 3.8 Hz, 1H), 7.62-7.56 (m, 1H).
MS (LR-APCI) calcd. for C.sub.15H.sub.11N.sub.2O.sub.2 (M+H) 251.1;
found 251.5.
3C: 2-Phenyl-[1,7]naphthyridine-4-carboxylic acid
##STR00040##
[0118] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (white
solid, 160 mg, 35%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
9.51 (s, 1H), 8.72 (s, 1H), 8.68 (d, J=6.4 Hz, 1H), 8.56 (d, J=6.4
Hz, 1H), 8.34-8.30 (m, 2H), 7.62-7.54 (m, 3H). MS (LR-APCI) calcd.
for C.sub.15H.sub.11N.sub.2O.sub.2 (M+H) 251.1; found 251.5.
3D: 2-Pyridin-2-yl-[1,7]naphthyridine-4-carboxylic acid
##STR00041##
[0120] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (77 mg,
34%). MS (LR-APCI) calcd. for C.sub.14H.sub.10N.sub.3O.sub.2 (M+H)
252.1; found 252.3.
3E: 2-(3-Methoxy-phenyl)-[1,7]naphthyridine-4-carboxylic acid
##STR00042##
[0122] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (70 mg,
28%). MS (LR-APCI) calcd. for C.sub.16H.sub.13N.sub.2O.sub.3 (M+H)
281.1; found 281.4.
3F: 2-Benzo[1,3]dioxol-5-yl-[1,7]naphthyridine-4-carboxylic
acid
##STR00043##
[0124] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (89 mg,
34%). MS (LR-APCI) calcd. for C.sub.16H.sub.11N.sub.2O.sub.4 (M+H)
295.1; found 295.5.
3G: 2-(3-Chloro-phenyl)-[1,7]naphthyridine-4-carboxylic acid
##STR00044##
[0126] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (tan
solid, 73 mg, 29%). MS (LR-APCI) calcd. for
C.sub.15H.sub.10ClN.sub.2O.sub.2 (M+H) 285.0; found 285.6.
3H: 2-Pyridin-2-yl-[1,8]naphthyridine-4-carboxylic acid
##STR00045##
[0128] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (gray
solid, 199 mg, 66%). MS (LR-APCI) calcd. for
C.sub.14H.sub.10N.sub.3O.sub.2 (M+H) 252.1; found 252.4.
3I: 2-(3-Methoxy-phenyl)-[1,8]naphthyridine-4-carboxylic acid
##STR00046##
[0130] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (yellow
solid, 252 mg, 75%). MS (LR-APCI) calcd. for
C.sub.16H.sub.13N.sub.2O.sub.3 (M+H) 281.1; found 281.4.
3J: 2-Benzo[1,3]dioxol-5-yl-[1,8]naphthyridine-4-carboxylic
acid
##STR00047##
[0132] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (golden
solid, 298 mg, 84%). MS (LR-APCI) calcd. for
C.sub.16H.sub.11N.sub.2O.sub.4 (M+H) 295.1; found 295.4.
3K: 2-(3-Chloro-phenyl)-[1,8]naphthyridine-4-carboxylic acid
##STR00048##
[0134] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (white
solid, 251 mg, 73%). MS (LR-APCI) calcd. for
C.sub.15H.sub.10ClN.sub.2O.sub.2 (M+H) 285.0; found 285.4.
3L: 2-Pyridin-2-yl-[1,6]naphthyridine-4-carboxylic acid
##STR00049##
[0136] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (gray
solid, 482 mg, 70%). MS (LR-APCI) calcd. for
C.sub.14H.sub.10N.sub.3O.sub.2 (M+H) 252.1; found 252.3.
3M: 2-(3-Methoxy-phenyl)-[1,6]naphthyridine-4-carboxylic acid
##STR00050##
[0138] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (yellow
solid, 548 mg, 72%). MS (LR-APCI) calcd. for
C.sub.16H.sub.13N.sub.2O.sub.3 (M+H) 281.1; found 281.4.
3N: 2-Benzo[1,3]dioxol-5-yl-[1,6]naphthyridine-4-carboxylic
acid
##STR00051##
[0140] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (golden
solid, 527 mg, 66%). MS (LR-APCI) calcd. for
C.sub.16H.sub.11N.sub.2O.sub.4 (M+H) 295.1; found 295.4.
3O: 2-(3-Chloro-phenyl)-[1,6]naphthyridine-4-carboxylic acid
##STR00052##
[0142] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (white
solid, 496 mg, 64%). MS (LR-APCI) calcd. for
C.sub.15H.sub.10ClN.sub.2O.sub.2 (M+H) 285.0; found 285.6.
3P: 2-m-Tolyl-[1,6]naphthyridine-4-carboxylic acid
##STR00053##
[0144] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (white
solid, 275 mg, 70%). MS (LR-APCI) calcd. for
C.sub.16H.sub.13N.sub.2O.sub.2 (M+H) 265.1; found 265.5.
3Q: 2-(3-Trifluoromethyl-phenyl)-[1,6]naphthyridine-4-carboxylic
acid
##STR00054##
[0146] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (white
solid, 247 mg, 71%). MS (LR-APCI) calcd. for
C.sub.16H.sub.10F.sub.3N.sub.2O.sub.2 (M+H) 319.1; found 319.5.
3R: 2-(4-Fluoro-phenyl)-[1,6]naphthyridine-4-carboxylic acid
##STR00055##
[0148] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (yellow
solid, 200 mg, 68%). MS (LR-APCI) calcd. for
C.sub.15H.sub.10FN.sub.2O.sub.2 (M+H) 269.1; found 269.5.
3S: 2-(4-Chloro-phenyl)-[1,6]naphthyridine-4-carboxylic acid
##STR00056##
[0150] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (white
solid, 82 mg, 26%). MS (LR-APCI) calcd. for
C.sub.15H.sub.10ClN.sub.2O.sub.2 (M+H) 285.0; found 285.5.
3T: 2-(4-Methoxy-phenyl)-[1,6]naphthyridine-4-carboxylic acid
##STR00057##
[0152] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (pale
yellow solid, 244 mg, 79%). MS (LR-APCI) calcd. for
C.sub.16H.sub.13N.sub.2O.sub.3 (M+H) 281.1; found 281.4.
3U: 2-p-Tolyl-[1,6]naphthyridine-4-carboxylic acid
##STR00058##
[0154] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (white
solid, 194 mg, 60%). MS (LR-APCI) calcd. for
C.sub.16H.sub.13N.sub.2O.sub.2 (M+H) 265.1; found 265.6.
3V: 2-(4-Trifluoromethyl-phenyl)-[1,6]naphthyridine-4-carboxylic
acid
##STR00059##
[0156] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics
(off-white solid, 270 mg, 69%). MS (LR-APCI) calcd. for
C.sub.16H.sub.10F.sub.3N.sub.2O.sub.2 (M+H) 319.1; found 319.6.
3W: 2-(3-Fluoro-phenyl)-[1,6]naphthyridine-4-carboxylic acid
##STR00060##
[0158] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (white
solid, 214 mg, 65%). MS (LR-APCI) calcd. for
C.sub.15H.sub.10FN.sub.2O.sub.2 (M+H) 269.1; found 269.5.
3X:
2-(2,3-Dihydro-benzo[1,4]dioxin-6-yl)-[1,6]naphthyridine-4-carboxylic
acid
##STR00061##
[0160] Following the procedure taught in Example 3A, the title
compound was prepared having the following characteristics (bright
yellow solid, 179 mg, 47%). MS (LR-APCI) calcd. for
C.sub.17H.sub.12N.sub.2O.sub.4 (M+H) 309.1; found 309.5.
Example 4
Preparation of 2-Aryl-[1,n]naphthyridin-4-ylamines
4A: 2-Phenyl-[1,8]naphthyridin-4-ylamine
##STR00062##
[0162] To a solution of 2-phenyl-[1,8]naphthyridine-4-carboxylic
acid (549 mg, 1.65 mmol) and triethylamine (0.314 mL, 1.4 equiv) in
DMF (to make a ca. 0.2 M solution) was added diphenylphosphoryl
azide (0.488 mL, 1.4 equiv) in one portion at ambient temperature
and the reaction was stirred for 3 h (note: the solution may become
heterogeneous but this will not adversely affect the reaction).
Water (add 0.15 mL for each 1.0 mL DMF used) was added and the
reaction heated to 100.degree. C. overnight. The reaction was
cooled to room temperature and poured into a vigorously stirring
mixture of 1N NaOH containing 1% conc. NH.sub.4OH. Stirred 15 min
and extracted with EtOAc. The organic layer was washed successively
with water and brine and was dried (MgSO.sub.4). The solution was
filtered, concentrated, and taken up in 100 mL EtOAc/Et.sub.2O
(1:1). Acidification with 2N HCl in Et.sub.2O afforded the product
as the HCl salt which was collected by vacuum filtration, rinsed
with EtOAc, and dried under high vacuum (light yellow solid, 215
mg). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.46 (s, 1H),
9.10-9.07 (m, 1H), 9.06-9.03 (m, 1H), 7.90-7.87 (m, 2H), 7.77 (dd,
J=8.6, 4.6 Hz, 1H), 7.69-7.60 (m, 3H), 7.08 (s, 1H). MS (LR-APCI)
calcd. for C.sub.14H.sub.12N.sub.3 (M+H) 222.1; found 222.5.
4B: 2-Phenyl-[1,7]naphthyridin-4-ylamine
##STR00063##
[0164] Following the procedure taught in Example 4a, the title
compound was prepared, as its HCl salt, having the following
characteristics (166 mg). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.60 (s, 1H), 9.46 (br s, 1H), 8.76 (d, J=6.4 Hz, 1H), 8.43
(d, J=6.0 Hz, 1H), 7.99-7.95 (m, 2H), 7.72-7.65 (m, 3H), 7.20 (s,
1H). MS (LR-APCI) calcd. for C.sub.14H.sub.12N.sub.3 (M+H) 222.1;
found 222.5.
4C: 2-Phenyl-[1,6]naphthyridin-4-ylamine
##STR00064##
[0166] Following the procedure taught in Example 4A, the title
compound was prepared, as its TFA salt, followed by preparative
reverse phase HPLC. The resultant compound had the following
characteristics, .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.73
(s, 1H), 9.36 (br s, 2H), 8.86 (d, J=6.4 Hz, 1H), 7.93-7.89 (m,
2H), 7.87 (d, J=6.4 Hz, 1H), 7.72-7.66 (m, 3H), 7.09 (s, 1H). MS
(LR-APCI) calcd. for C.sub.14H.sub.12N.sub.3 (M+H) 222.1; found
222.5.
4D: 2-(3-Chloro-phenyl)-[1,7]naphthyridin-4-ylamine
##STR00065##
[0168] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (orange solid, 61 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.58 (s, 1H), 9.46 (br s, 2H), 8.85-8.79 (m,
1H), 8.50-8.44 (m, 1H), 8.13 (s, 1H), 7.99-7.93 (m, 1H), 7.85-7.73
(m, 2H), 7.25 (s, 1H). MS (LR-APCI) calcd. for
C.sub.14H.sub.11ClN.sub.3 (M+H) 256.1; found 256.3.
4E: 2-Pyridin-2-yl-[1,8]naphthyridin-4-ylamine
##STR00066##
[0170] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (yellow solid, 57 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.55 (br s, 2H), 9.16-9.13 (m, 1H), 9.09-9.06
(m, 1H), 8.90-8.87 (m, 1H), 8.38 (d, J=7.6 Hz, 1H), 8.18-8.13 (m,
1H), 7.82 (dd, J=8.2, 4.2 Hz, 1H), 7.72 (dd, J=7.8, 4.6 Hz, 1H),
7.63 (s, 1H). MS (LR-APCI) calcd. for C.sub.13H.sub.11N.sub.4 (M+H)
223.1; found 223.2.
4F: 2-(3-Methoxy-phenyl)-[1,8]naphthyridin-4-ylamine
##STR00067##
[0172] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (yellow solid, 210 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.46 (br s, 2H), 9.13-9.10 (m, 1H), 9.07-9.03
(m, 1H), 7.80 (dd, J=8.2, 4.2 Hz, 1H), 7.57 (t, J=7.8 Hz, 1H),
7.51-7.49 (m, 1H), 7.46 (d, J=8.0 Hz, 1H), 7.27-7.23 (m, 1H), 7.12
(s, 1H), 3.89 (s, 3H). MS (LR-APCI) calcd. for
C.sub.15H.sub.14N.sub.3O (M+H) 252.1; found 252.4.
4G: 2-Benzo[1,3]dioxol-5-yl-[1,8]naphthyridin-4-ylamine
##STR00068##
[0174] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (dark orange solid, 56 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.38 (br s, 2H), 9.10-9.07 (m, 1H), 9.05-9.02
(m, 1H), 7.77 (dd, J=8.4, 4.8 Hz, 1H), 7.52-7.51 (m, 1H), 7.49-7.45
(m, 1H), 7.20 (d, J=8.0 Hz, 1H), 7.06 (s, 1H), 6.20 (s, 2H). MS
(LR-APCI) calcd. for C.sub.15H.sub.12N.sub.3O.sub.2 (M+H) 266.1;
found 266.5.
4H: 2-(3-Chloro-phenyl)-[1,8]naphthyridin-4-ylamine
##STR00069##
[0176] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (yellow solid, 203 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.46 (br s, 2H), 9.14-9.12 (m, 1H), 9.07-9.03
(m, 1H), 8.02 (s, 1H), 7.86 (d, J=7.6 Hz, 1H), 7.81 (dd, J=8.2, 4.2
Hz, 1H), 7.76 (d, J=7.6 Hz, 1H), 7.68 (t, J=7.8 Hz, 1H), 7.09 (s,
1H). MS (LR-APCI) calcd. for C.sub.14H.sub.11ClN.sub.3 (M+H) 256.1;
found 256.3.
4I: 2-Pyridin-2-yl-[1,6]naphthyridin-4-ylamine
##STR00070##
[0178] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (yellow solid, 65 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.90 (s, 1H), 9.72 (br s, 2H), 8.93-8.91 (m,
1H), 8.88 (d, J=6.0 Hz, 1H), 8.40 (d, J=7.6 Hz, 1H), 8.24 (d, J=6.0
Hz, 1H), 8.20 (td, J=7.6, 1.6 Hz, 1H), 7.78-7.74 (m, 1H), 7.72 (s,
1H). MS (LR-APCI) calcd. for C.sub.13H, 1N.sub.4 (M+H) 223.1; found
223.3.
4J: 2-(3-Methoxy-phenyl)-[1,6]naphthyridin-4-ylamine
##STR00071##
[0180] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (light orange solid, 211 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.83 (s, 1H), 9.59 (br s, 2H), 8.88 (d, J=6.4
Hz, 1H), 8.06 (d, J=6.4 Hz, 1H), 7.61 (t, J=8.0 Hz, 1H), 7.54 (s,
1H), 7.50 (d, J=8.4 Hz, 1H), 7.31-7.27 (m, 1H), 7.17 (s, 1H), 3.90
(s, 3H). MS (LR-APCI) calcd. for C.sub.15H.sub.14N.sub.3O (M+H)
252.1; found 252.4.
4K: 2-Benzo[1,3]dioxol-5-yl-[1,6]naphthyridin-4-ylamine
##STR00072##
[0182] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (dark orange solid, 230 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.90 (s, 1H), 9.62 (br s, 2H), 8.87 (d, J=5.2
Hz, 1H), 8.16 (d, J=6.4 Hz, 1H), 7.61 (s, 1H), 7.56 (d, J=8.8 Hz,
1H), 7.23 (d, J=7.6 Hz, 1H), 7.17 (s, 1H), 6.22 (s, 2H). MS
(LR-APCI) calcd. for C.sub.15H.sub.12N.sub.3O.sub.2 (M+H) 266.1;
found 266.4.
4L: 2-(3-Chloro-phenyl)-[1,6]naphthyridin-4-ylamine
##STR00073##
[0184] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (light orange solid, 255 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.88 (s, 1H), 9.57 (br s, 2H), 8.91-8.85 (m,
1H), 8.09 (s, 1H), 8.08-8.02 (m, 1H), 7.97-7.90 (1H), 7.81-7.68 (m,
2H), 7.21 (s, 1H). MS (LR-APCI) calcd. for
C.sub.14H.sub.11ClN.sub.3 (M+H) 256.1; found 256.5.
4M: 2-m-Tolyl-[1,6]naphthyridin-4-ylamine
##STR00074##
[0186] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.86
(s, 1H), 9.62 (br s, 2H), 8.88 (d, J=5.2 Hz, 1H), 8.06 (d, J=6.0
Hz, 1H), 7.80 (s, 1H), 7.75 (d, J=8.0 Hz, 1H), 7.58 (t, J=7.2 Hz,
1H), 7.54 (d, J=8.0 Hz, 1H), 7.17 (s, 1H), 2.46 (s, 3H). MS
(LR-APCI) calcd. for C.sub.15H.sub.14N.sub.3 (M+H) 236.1; found
236.6.
4N: 2-(3-Trifluoromethyl-phenyl)-[1,6]naphthyridin-4-ylamine
##STR00075##
[0188] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (245 mg). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.90 (s, 1H), 9.51 (br s, 2H), 8.88 (d, J=6.0 Hz, 1H), 8.36
(s, 1H), 8.29 (d, J=8.0 Hz, 1H), 8.10-8.04 (m, 2H), 7.92 (t, J=7.8
Hz, 1H), 7.26 (s, 1H). MS (LR-APCI) calcd. for
C.sub.15H.sub.11F.sub.3N.sub.3 (M+H) 290.1; found 290.6.
4O: 2-(4-Fluoro-phenyl)-[1,6]naphthyridin-4-ylamine
##STR00076##
[0190] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (163 mg). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.86 (s, 1H), 9.57 (br s, 2H), 8.87 (d, J=5.6 Hz, 1H),
8.08-8.03 (m, 3H), 7.56 (t, J=9.2 Hz, 2H), 7.16 (s, 1H). MS
(LR-APCI) calcd. for C.sub.14H.sub.11FN.sub.3 (M+H) 240.1; found
240.2.
4P: 2-(4-Chloro-phenyl)-[1,6]naphthyridin-4-ylamine
##STR00077##
[0192] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (23 mg). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.87 (s, 1H), 9.58 (br s, 2H), 8.87 (d, J=5.2 Hz, 1H), 8.06
(d, J=5.2 Hz, 1H), 8.01 (d, J=8.4 Hz, 2H), 7.78 (d, J=8.4 Hz, 2H),
7.18 (s, 1H). MS (LR-APCI) calcd. for C.sub.14H.sub.11ClN.sub.3
(M+H) 256.1; found 256.5.
4Q: 2-(4-Methoxy-phenyl)-[1,6]naphthyridin-4-ylamine
##STR00078##
[0194] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (154 mg). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.91 (s, 1H), 9.65 (br s, 2H), 8.87 (d, J=6.0 Hz, 1H), 8.20
(d, J=6.4 Hz, 1H), 8.01 (d, J=9.6 Hz, 2H), 7.24 (d, J=8.8 Hz, 2H),
7.21 (s, 1H), 3.90 (s, 3H). MS (LR-APCI) calcd. for
C.sub.15H.sub.14N.sub.3O (M+H) 252.1; found 252.4.
4R: 2-p-Tolyl-[1,6]naphthyridin-4-ylamine
##STR00079##
[0196] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (dirty pale yellow solid, 136 mg). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.92 (s, 1H), 9.71 (br s, 2H), 8.88 (d,
J=6.4 Hz, 1H), 8.18 (d, J=6.4 Hz, 1H), 7.91 (d, J=8.0 Hz, 2H), 7.50
(d, J=8.8 Hz, 2H), 7.22 (s, 1H), 2.44 (s, 3H). MS (LR-APCI) calcd.
for C.sub.15H.sub.14N.sub.3 (M+H) 236.1; found 236.6.
4S: 2-(4-Trifluoromethyl-phenyl)-[1,6]naphthyridin-4-ylamine
##STR00080##
[0198] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (tan solid, 158 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.99 (s, 1H), 9.76 (br s, 2H), 8.88 (d, J=6.4
Hz, 1H), 8.24-8.18 (m, 3H), 8.06 (d, J=8.4 Hz, 2H), 7.29 (s, 1H).
MS (LR-APCI) calcd. for C.sub.15H.sub.11F.sub.3N.sub.3 (M+H) 290.1;
found 290.6.
4T: 2-(3-Fluoro-phenyl)-[1,6]naphthyridin-4-ylamine
##STR00081##
[0200] Following the procedure taught in Example 4A, the title
compound was prepared as its HCl salt. The crude HCl salt was
further purified by C18 chromatography to obtain the product as its
free base having the following characteristics (bright yellow
solid, 71 mg). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.89 (s,
1H), 9.62 (br s, 2H), 8.84 (d, J=6.0 Hz, 1H), 8.12 (d, J=5.2 Hz,
1H), 7.91 (d, J=10.0 Hz, 1H), 7.84 (d, J=7.6 Hz, 1H), 7.74-7.68 (m,
1H), 7.54 (td, J=8.2, 1.6 Hz, 1H), 7.27 (s, 1H). MS (LR-APCI)
calcd. for C.sub.14H.sub.11FN.sub.3 (M+H) 240.1; found 240.5.
4U:
2-(2,3-Dihydro-benzo[1,4]dioxin-6-yl)-[1,6]naphthyridin-4-ylamine
##STR00082##
[0202] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (yellow solid, 170 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.90 (s, 1H), 9.65 (br s, 2H), 8.87 (d, J=6.4
Hz, 1H), 8.16 (d, J=6.4 Hz, 1H), 7.59 (d, J=2.4 Hz, 1H), 7.51 (dd,
J=8.8, 1.6 Hz, 1H), 7.19 (s, 1H), 7.16 (d, J=8.8 Hz, 1H), 4.37 (dd,
J=10.0, 5.2 Hz, 4H). MS (LR-APCI) calcd. for
C.sub.16H.sub.14N.sub.3O.sub.2 (M+H) 280.1; found 280.4.
4V: 2-Pyridin-2-yl-[1,7]naphthyridin-4-ylamine
##STR00083##
[0204] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (62 mg). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.84 (s, 1H), 9.72 (br s, 2H), 8.92 (d, J=3.6 Hz, 1H), 8.77
(d, J=5.2 Hz, 1H), 8.56 (d, J=6.0 Hz, 1H), 8.48 (d, J=8.0 Hz, 1H),
8.20 (td, J=8.0, 1.6 Hz, 1H), 7.81 (s, 1H), 7.76 (dd, J=7.6, 4.8
Hz, 1H). MS (LR-APCI) calcd. for C.sub.13H, 1N.sub.4 (M+H) 223.1;
found 223.4.
4W: 2-(3-Methoxy-phenyl)-[1,7]naphthyridin-4-ylamine
##STR00084##
[0206] Following the procedure taught in Example 4A, the title
compound was prepared, as its HCl salt, having the following
characteristics (65 mg). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.75 (s, 1H), 9.55 (br s, 2H), 8.77 (d, J=5.2 Hz, 1H), 8.49
(d, J=6.0 Hz, 1H), 7.62 (s, 1H), 7.59 (d, J=7.6 Hz, 1H), 7.53 (d,
J=7.6 Hz, 1H), 7.29-7.25 (m, 2H), 3.92 (s, 3H). MS (LR-APCI) calcd.
for C.sub.15H.sub.14N.sub.3O (M+H) 252.1; found 252.4.
4X: 2-Benzo[1,3]dioxol-5-yl-[1,7]naphthyridin-4-ylamine
##STR00085##
[0208] Following the procedure taught in Example 4A, the title
compound was prepared, as its TFA salt. The TFA salt was further
purified by preparative reverse phase HPLC, yielding a compound
having the following characteristics. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.39 (s, 1H), 9.01 (br s, 2H), 8.74 (d, J=6.4
Hz, 1H), 8.30 (d, J=5.6 Hz, 1H), 7.56 (d, J=1.6 Hz, 1H), 7.53-7.50
(m, 1H), 7.24 (d, J=7.6 Hz, 1H), 7.11 (s, 1H), 6.21 (s, 2H). MS
(LR-APCI) calcd. for C.sub.15H.sub.12N.sub.3O.sub.2 (M+H) 266.1;
found 266.5.
Example 5
Functional Mobilization of Intracellular Calcium to Determine
Muscarinic Receptor Activity
[0209] CHO cells expressing muscarinic receptors (M1 to M5) are
grown as monolayers in tissue culture flasks at 37.degree. C. in a
humidified atmosphere containing 5% CO.sub.2 and passaged every 3-5
days. The growth media is Dulbecco's modified eagles medium (DMEM,
Gibco Cat# 12430-054), containing 25 mM Hepes and supplemented with
Fetal Bovine Serum (Hyclone, cat# SH30071.03), 0.1 mM of MEM
non-essential amino acids (GIBCO, Cat# 11140-050), 1 mM MEM Sodium
Pyruvate (GIBCO Cat# 11360-070) and 100 units/ml of Penicillin G
and 100 .mu.g/ml of Streptomycin (GIBCO Cat# 15140-122). The
recombinant muscarinic receptor cell lines are grown under
antibiotic pressure with media containing 25 .mu.g/ml zeocin and
500 .mu.g/ml G418 (M1-CHO), 4 .mu.g/ml puromycin, 50 .mu.g/ml
zeocin and 2.5 .mu.g/ml blasticidin (M2 and M4-CHO) or 50 .mu.g/ml
zeocin and 4 .mu.g/ml puromycin (M3 and M5-CHO).
[0210] Cells are harvested at 80-90% confluence using Versene
(GIBCO Cat# 15040-066), collected by centrifugation and seeded
18-24 hrs prior to running the calcium assay at a density of
5,000-10,000 cells/well in back-walled, clear-bottomed 384-well
plates (BD Biocoat, poly-D-lysine, Cat#356663). The day of the
experiment, the cells are washed with a plate washer (Bioteck
Instruments, ELX 405) using bath 1 buffer (140-mM NaCl, 4.5-mM KCl,
2-mM CaCl.sub.2, 1-mM MgCl.sub.2, 10-mM Hepes-Na, 10-mM Glucose, pH
7.4, with NaOH) containing 1 mM Probenecid. Next, the calcium dye
Fluo-3 (25 .mu.l/well of Fluo-3 AM at 4 .mu.M, Molecular Probes
F-1241, in Bath 1 buffer containing 1 mM Probenecid) is added to
the 25 .mu.l of Bath I remaining in each well after the plate wash
and the dye is loaded at 37.degree. C. in the tissue culture
incubator for 60-90 min. The fluorescent dye is removed using the
plate washer with Bath I containing 1 mM Probenecid, leaving 25
.mu.l/well of this solution after the wash. Alternatively, cells
can be loaded with the calcium indicator from Molecular Devices
(Calcium 3 Assay Reagents, Cat # R7181) adding 5 .mu.l of a
5.times. solution dye in Bath 1 containing 1 mM Probenecid (10 ml
per dye flask cat# R7182 to generate a solution 20.times.) to 20
.mu.l of the same buffer. After loading for 60 min, the experiment
can be run without having to remove the dye.
[0211] Compounds are prepared at a 2.times. fold concentration in a
96-well plate (round bottom, Costar Corning cat# 3656), by
reconstituting the pre-spotted compounds in bath I containing 1 mM
probenecid. The final concentration DMSO is 0.5%, and the amount of
DMSO is normalized across the assay plate. To determine an agonist
action of the compounds on muscarinic receptors, the reconstituted
compounds are added (25 .mu.l compound/well) to the cell assay
plate (containing 25 .mu.l/well) using the multi-channel robotic
system of the FLIPR 3 Instrument (Molecular Devices, Sunnyvale,
Calif.). To determine a functional inhibitory action of the
compounds on muscarinic receptors, the reconstituted compounds are
added (25 .mu.l compound/well) to the assay plate and pre-incubated
for 15 min prior to adding 25 .mu.l of Carbachol at 3.times. the
EC80 for each muscarinic subtype. Alternatively, the compounds can
be co-applied simultaneously with the agonist. In both assay modes,
the fluorescence is recorded for 60 sec (excitation wavelength is
488 nM and emission wavelength 540 nm) using the FLIPR 3
instrument.
[0212] The potency, efficacy and selectivity of the muscarinic
compounds were evaluated by screening the compound activity across
the whole family (M1 to M5 cells).
[0213] The compounds of the present invention were found to
selectively modulate the muscarinic receptors selectively over the
other receptor types.
Example 6
.beta.-Lactamase Assay to Determine Muscarinic Receptor
Activity
[0214] CHO cells expressing muscarinic receptors (M1 to M5) and
containing a gene reporter system (.beta.-Lactamase) with
transcriptional control mediated by calcium release (NFAT
activation). See Zlokarnik, G; Negulescu, P. A.; Knapp, T. E.;
Mere, L; Burres, N; Feng, L; Whitney, M; Roemer, K; Tsien, R. Y.
Quantitation of transcription and clonal selection of single living
cells with .beta.-lactamase as reporter. Science, 1998 Jan. 2,
279(5347):84-8. The cells are grown as monolayers in tissue culture
flasks at 37.degree. C. in a humidified atmosphere containing 5%
CO.sub.2 and passaged every 3-5 days. The growth media is
Dulbecco's modified eagles medium (DMEM, Gibco Cat# 12430-054),
containing 25 mM Hepes and supplemented with 10% Fetal Bovine Serum
(Hyclone, cat# SH30071.03), 0.1 mM of MEM non-essential amino acids
(GIBCO, Cat# 11140-050), 1 mM MEM Sodium Pyruvate (GIBCO Cat#
11360-070) and 100 units/ml of Penicillin G and 100 .mu.g/ml of
Streptomycin (GIBCO Cat# 15140-122). The recombinant muscarinic
receptor cell lines are grown under antibiotic pressure with media
containing 25 .mu.g/ml zeocin and 500 .mu.g/ml G418 (M1-CHO), 4
.mu.g/ml puromycin, 50 .mu.g/ml zeocin and 2.5 .mu.g/ml blasticidin
(M2 and M4-CHO) or 50 .mu.g/ml zeocin and 4 .mu.g/ml puromycin (M3
and M5-CHO).
[0215] Cells are harvested at 80-90% confluence using Accutase
(Innovative Cell Technologies, Inc. Cat# AT104), collected by
centrifugation and seeded for 2-6 hours at a density of
15,000-20,000 cells/well in black-walled, clear-bottomed 384-well
plates (BD Biocoat, poly-D-lysine, Cat#356663). Media is replaced
with DMEM+1% Fetal Bovine Serum and incubated for another 12-18 hrs
prior to running the .beta.-Lactamase assay. The day of the
experiment, compounds are prepared at a 1.times. fold concentration
in a 96-well plate (round bottom, Costar Corning cat# 3656), by
reconstituting the pre-spotted compounds in DMEM+1% FBS. The final
concentration of DMSO is 0.5%, and the amount of DMSO is normalized
across the assay plate. To determine an agonist action of the
compounds on muscarinic receptors, the reconstituted compounds are
added (25 .mu.l compound/well) to the cell assay plate (where the
media has been removed) using the multi-channel robotic system,
Multimek 96 (Beckman). The compounds are incubated with the cells
for 3 hours at 37.degree. C., 5% CO.sub.2. to allow for expression
of the reporter gene .beta.-Lactamase. After 3 hours, 5 .mu.l of
6.times. fold concentrated CCF2/AM dye are added to the assay
plates and incubated at room temperature for 1 hour. Fluorescent
emission at two wavelengths (460 nm and 530 nm) is determined using
the CytoFluor Series 4000 (PerSeptive Biosystems) and the
calculations for reporter gene expression determined as specified
in prior publications {Zlokarnik, G; Negulescu, P. A.; Knapp, T.
E.; Mere, L; Burres, N; Feng, L; Whitney, M; Roemer, K; Tsien, R.
Y. Quantitation of transcription and clonal selection of single
living cells with .beta.-lactamase as reporter. Science, 1998 Jan.
2, 279(5347):84-8.}
[0216] The compounds of the present invention were found to
modulate the muscarinic receptor activity using the
.beta.-Lactamase Assay.
* * * * *