U.S. patent application number 13/271028 was filed with the patent office on 2012-02-09 for quinuclidine compounds as alpha-7 nicotinic acetylcholine receptor ligands.
This patent application is currently assigned to Bristol-Myers Squibb Company. Invention is credited to James H. Cook, II, Christiana I. Iwuagwu, Dalton King, John E. Macor, Ivar M. McDonald, Richard E. Olson, Nenghui Wang, F. Christopher Zusi.
Application Number | 20120035189 13/271028 |
Document ID | / |
Family ID | 42109161 |
Filed Date | 2012-02-09 |
United States Patent
Application |
20120035189 |
Kind Code |
A1 |
Cook, II; James H. ; et
al. |
February 9, 2012 |
Quinuclidine Compounds as Alpha-7 Nicotinic Acetylcholine Receptor
Ligands
Abstract
The disclosure provides compounds of formula I, including their
salts, as well as compositions and methods of using the compounds.
The compounds are ligands for the nicotinic .alpha.7 receptor and
may be useful for the treatment of various disorders of the central
nervous system, especially affective and neurodegenerative
disorders. ##STR00001##
Inventors: |
Cook, II; James H.; (East
Hampton, CT) ; McDonald; Ivar M.; (East Haddam,
CT) ; King; Dalton; (Hamden, CT) ; Olson;
Richard E.; (Orange, CT) ; Wang; Nenghui;
(Guilford, CT) ; Iwuagwu; Christiana I.; (Hamden,
CT) ; Zusi; F. Christopher; (Hamden, CT) ;
Macor; John E.; (Guilford, CT) |
Assignee: |
Bristol-Myers Squibb
Company
|
Family ID: |
42109161 |
Appl. No.: |
13/271028 |
Filed: |
October 11, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12607354 |
Oct 28, 2009 |
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13271028 |
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12423299 |
Apr 14, 2009 |
7863291 |
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12607354 |
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61047211 |
Apr 23, 2008 |
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Current U.S.
Class: |
514/256 ;
544/230 |
Current CPC
Class: |
C07D 498/20 20130101;
C07D 519/00 20130101; A61P 3/10 20180101; A61P 25/18 20180101; A61P
19/02 20180101; A61K 31/439 20130101; A61P 29/00 20180101; A61P
3/00 20180101; A61K 31/506 20130101; A61P 25/28 20180101; A61P 1/00
20180101 |
Class at
Publication: |
514/256 ;
544/230 |
International
Class: |
A61K 31/506 20060101
A61K031/506; A61P 25/28 20060101 A61P025/28; A61P 3/00 20060101
A61P003/00; A61P 19/02 20060101 A61P019/02; A61P 1/00 20060101
A61P001/00; C07D 413/14 20060101 C07D413/14; A61P 25/18 20060101
A61P025/18 |
Claims
1. A compound of formula I, or a stereoisomer thereof, ##STR01098##
wherein R.sup.1 is pyrimidinyl substituted with 1 imidazolyl
substituent; or a pharmaceutically acceptable salt thereof.
2. The compound of claim 1:
(R)--N-(6-(1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine ##STR01099## or a pharmaceutically
acceptable salt thereof.
3. A pharmaceutical composition comprising a therapeutically
effective amount of a compound of claim 1, or a pharmaceutically
acceptable salt thereof, and a pharmaceutically acceptable
carrier.
4. The composition of claim 3 where the compound of claim 1 is
(R)--N-(6-(1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine ##STR01100## or a pharmaceutically
acceptable salt thereof, and a pharmaceutically acceptable
carrier.
5. A method for the treatment of schizophrenia, Alzheimer's
Disease, cognitive disorders, rheumatoid arthritis, osteoarthritis,
ulcerative colitis, Crohn's Disease, or diabetes comprising
administering a therapeutically affective amount of a compound of
claim 1 to a patient.
6. The method of claim 5 directed to schizophrenia.
7. The method of claim 6 where the compound of claim 1 is
(R)--N-(6-(1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine ##STR01101## or a pharmaceutically
acceptable salt thereof.
8. The method of claim 7 directed to Alzheimer's disease.
9. The method of claim 8 where the compound of claim 1 is
(R)--N-(6-(1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine ##STR01102## or a pharmaceutically
acceptable salt thereof.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This patent application is a Continuation of
Continuation-in-Part Application U.S. Ser. No. 12/607,354 filed
Oct. 28, 2009 which claims the benefit of U.S. Non-Provisional
Application U.S. Ser. No. 12/423,299 filed Apr. 14, 2009, now U.S.
Pat. No. 7,863,291 which claims the benefit of U.S. Provisional
Application U.S. Ser. No. 61/047,211 filed Apr. 23, 2008.
BACKGROUND OF THE INVENTION
[0002] The disclosure generally relates to compounds of formula I,
including their salts, as well as compositions and methods of using
the compounds. The compounds are ligands, agonists and partial
agonists for the nicotinic .alpha.7 receptor and may be useful for
the treatment of various disorders of the central nervous system,
especially affective and neurodegenerative disorders.
[0003] Schizophrenia is a serious mental disorder, affecting
approximately 1% of the population. Its progressive course results
in major impairment of mental and social functioning and often
leads to the development of other pathologies. Susceptibility often
runs in families, with both genetic and environmental factors
thought to be important. The direct and indirect costs of the
disease are estimated in the tens of billion dollars annually in
the U.S. alone.
[0004] Patients with schizophrenia have an elevated risk of suicide
(approximately a 10% lifetime risk). They have a 2.5 fold increase
in all-cause mortality, resulting in a 20% lowered life expectancy.
The onset of illness can result in cascade of unhealthy lifestyle
factors and behaviors that elevate the risk of various conditions
and consequently the risk of death.
[0005] The onset of schizophrenia is most often in late adolescence
or early adulthood, and episodes recur throughout life. The disease
is characterized by the expression of three distinct symptom
domains: positive, negative and cognitive. Psychotic or positive
symptoms include delusions, hallucinations, thought disorder and
paranoia. Negative symptoms include negative affect, social
withdrawal, and anhedonia. Cognitive dysfunction includes deficits
in attention, working memory and executive function. The
pathophysiology of schizophrenia is not well understood, however,
most experts believe it is a multi-factorial disorder in which
biological, genetic and environmental factors play a role. Most
current therapies target the dopaminergic system and have resulted
in the suggestion that an excess of dopaminergic neurotransmission
underlies at least some aspects of schizophrenia. This theory
received further support from findings that drugs which increase
the levels of dopamine cause psychoses similar to the positive
symptoms of the disease. Also, post mortem analysis of brains from
schizophrenic patients indicate increased numbers of D2 dopamine
receptors. Although newer antipsychotic agents, known as atypical
antipsychotics, which are active at several additional
neurotransmitter receptors, have been introduced in the past
decade, these agents still share efficacy against the D2 dopamine
receptor. All currently-used agents also have major limitations.
Although positive symptoms are generally reduced in a majority of
patients, these drugs do little to relieve the negative symptoms
and cognitive deficits that are common and often most debilitating.
In addition, antipsychotic agents have a number of unwanted and
limiting side effects.
[0006] Nicotine is among the few agents which have a positive
effect on cognitive function. Many schizophrenics smoke; the rate
in patients is 2-4 times that of the general population, and up to
90% in schizophrenics who have been institutionalized do smoke.
This smoking habit has been characterized as a form of
self-medication.
[0007] Nicotinic acetylcholine receptors (nAChR's) are pentameric
ligand-gated ion channels which are widely expressed through the
central and peripheral nervous system. These channels are
fast-desensitizing calcium channels which, when open, increase the
intracellular concentration of the Ca.sup.++ ion. Although there
are 12 individual receptors, the most abundant nicotinic receptors
in the brain are .alpha.4.beta.2 and .alpha.7. The .alpha.4.beta.2
complex has been identified as the "high affinity" nicotine site.
The homo-pentameric .alpha.7 receptor selectively binds the natural
product, .alpha.-bungarotoxin, which has allowed its relatively
facile localization and measurement. The .alpha.7 receptor is
primarily expressed in the cortex, hippocampus and subcortical
limbic regions and commonly occurs pre-synaptically. The
localization of .alpha.7 nAChRs in areas involved with learning and
memory has led to studies using both knockout mice and
pharmacological manipulation. It is involved in sensory gating,
memory, and neuronal plasticity. Alpha7 agonists have been shown to
increase the release of neurotransmitters in rodents, including
dopamine, serotonin, glutamate and GABA. Compounds which
selectively bind to the .alpha.7 receptor, such as .alpha.7
agonists and partial agonists, have been shown to improve learning
and memory functions in normal and aged animals, reverse
scopolamine-induced memory deficits, reverse deficits in cognition
induced by NMDA antagonists, reverse pharmacologically-induced
gating deficits, e.g. amphetamine induced gating disruption, and to
possess some anxiolytic properties. The .alpha.7 agonists of the
present invention are expected to be useful in the treatment of
schizophrenia and cognitive disorders associated with
schizophrenia.
[0008] Alzheimer's disease is a progressive neurodegenerative
disorder, resulting in the general loss of cognitive functions. The
incidence increases with age, to the degree that 25-50% of all
individuals over 85 are estimated to suffer from some degree of
dementia. A diagnosis of Alzheimer's implies that the remaining
life expectancy is reduced by half, compared to normal adults.
[0009] Clinical signs of Alzheimer's disease are progressive
cognitive deterioration, decreased ability to perform the
activities of daily living and neuropsychiatric symptoms or
behavioral changes. In the advanced stages of the disease,
deterioration of musculature and mobility may lead to inability to
feed oneself, and eventually to the patient becoming bedridden.
Language becomes severely disorganized, and then is lost
altogether. Patients are not able to perform even simple tasks
independently and require constant supervision. The cost of
institutional care makes up nearly 70% of the cost of the disease.
Therefore, therapies which increase cognitive function and delay
institutionalization are greatly needed.
[0010] Alzheimer's disease has been shown in several studies to be
accompanied by a reduction in nicotinic receptors in the cortex and
hippocampus. Nicotine injections or nicotine skin patches have been
reported to significantly improve attention, memory and learning in
Alzheimer's disease patients. While there is a progressive loss of
nicotinic receptors during the course of Alzheimer's disease, the
.alpha.7 neurons are relatively spared, compared to the more
abundant .alpha.4 receptors. Recently, the administration of
selective nicotinic .alpha.7 agonists has been shown to increase
cognitive functioning in Alzheimer's patients when dosed as long as
8 weeks. This clinical data is consistent with pre-clinical data
showing .alpha.7 agonists and partial agonists improve learning and
memory functions in normal and aged animals and reverse
scopolamine-induced memory deficits. Thus, the compounds of the
present invention may be useful in the treatment and prevention of
Alzheimer's disease. The amyloid peptide A.beta.2 has been shown to
bind to the .alpha.7 nicotinic receptor (Wang et al., J. Biol.
Chem., 2000, 275:5626-5632; J. Neurochem. 2000, 75:1155-1161). This
association may facilitate the aggregation of A.beta.2, believed to
be important in the toxic effects of A.beta.2, and may also cause
disregulation of signaling through .alpha.7 nicotinic receptors.
Deletion of the .alpha.7 receptor gene improves cognitive deficits
and synaptic pathology in a mouse model of Alzheimer's disease
(Dziewczapolski et al., J. Neuroscience, 2009, pp 8805-8815). The
compounds of the present invention may disrupt the interaction of
A.beta.2 and .alpha.7 receptors. Treatment with .alpha.7 agonists
and partial agonists may represent an approach for disease
modification in Alzheimer's disease. Alpha7 receptors may also
mediate inflammatory processes in neurodegenerative conditions,
such as Alzheimer's disease (Conejero-Goldberg et al., Neurosci.
and Biobehav. Rev., 2008, 32, pp 693-706). The .alpha.7 agonists
and partial agonists of the present invention may be useful in
reducing inflammation in neurodegenerative diseases and disorders,
such as Alzheimer's disease.
[0011] The .alpha.7 receptor has also been shown to be involved in
the reduction of inflammation via the vagus nerve. In addition, the
.alpha.7 receptor is expressed in synoviocytes from RA and OA
patients, and .alpha.7 agonists have been shown to inhibit the
proinflammatory cascade that occurs in the rheumatoid joint
(Waldberger et al., Arthritis and Rheumatism, Vol 58, pp
3439-3449). Thus, the compounds of the present invention may be
useful in the treatment of inflammatory conditions, such as
rheumatoid arthritis and osteoarthritis.
[0012] Nicotinic receptors containing the .alpha.7 subunit are
present on mucosal mast cells known to be involved in
gastrointestinal hypersensitivity (Kageyama-Yahara et al., Biochem
and Biophys. Research Commun., 2008, v. 377, pp 321-325). The
.alpha.7 agonist GTS-21 inhibits the antigen-induced degranulation
of mucosal mast cells, suggesting that .alpha.7 agonists may be
useful in the treatment of hypersensitive bowel conditions, such as
ulcerative colitis.
[0013] In a recent report (Marrero et al., JPET Fast Forward, Sep.
28, 2009, DOI: 10.1124/jpet.109.154633), an .alpha.7 agonist was
shown to decrease weight gain and food intake and reduce the
elevated plasma levels of triglycerides, glucose, glycated
hemoglobin and TNFa in a mouse model of type II diabetes (db/db
mice which are deficit in leptin receptors). The .alpha.7 agonists
and partial agonists of the present invention may be useful in the
treatment of diabetes.
[0014] The following references provide general reviews of the
nicotinic receptor system and .alpha.7 receptors and ligands:
Picciotto and Zoli, J. Neurobio. (2002) 53:641-655; Brening, et al,
Ann. Reports in Med. Chem. (2005) 40:3-16; Dani and Bertrand, Ann.
Rev. Pharm. Tox. (2007) 47:699-729; Olincy and Stevens, Biochem.
Pharmacol. (2007) 74:1192-1201; Broad, et al, Drugs Future (2007)
32 (2):161-70; de Jonge and Ulloa, Brit. J. Pharmacol. (2007)
151:915-929; Romanelli, et al, ChemMedChem (2007) 2(6):746-767;
Lightfoot et al., Progress in Medicinal Chemistry (2008), v 46, pp
131-171; Concotta et al., Current Opinion in Investigational Drugs
(2008), v 9, pp 47-56; Leiser et al., Pharmacol. and Therapeutics
(2009), doi:10:1016/j.pharmthera.2009.03.009).
[0015] Ligands for the nicotinic .alpha.7 receptor have been
disclosed in the references above, and also in US patent
application publication U.S. 2007004715, WO 2008/000469, WO
2003/092580, WO 2004/000,469, EP 337,547, EP 452,101, and C. J.
Swain, et al., J. Med. Chem., (1992) 35:1019-1031.
[0016] The invention provides technical advantages, for example,
the compounds are novel and are ligands for the nicotinic .alpha.7
receptor and may be useful for the treatment of various disorders
of the central nervous system, especially affective and
neurodegenerative disorders. Additionally, the compounds provide
advantages for pharmaceutical uses, for example, with regard to one
or more of their mechanism of action, binding, inhibition efficacy,
target selectivity, solubility, safety profiles, or
bioavailability.
DESCRIPTION OF THE INVENTION
[0017] The invention encompasses compounds formula I, including
pharmaceutically acceptable salts, and compositions and methods of
treatment using these compounds. The compounds may be useful for
the treatment of various disorders of the central nervous
system:
[0018] One aspect of the invention is a compound of formula I, or a
stereoisomer thereof,
##STR00002##
wherein: R.sup.1 is selected from the group consisting of
isoxazolyl, pyrazolyl, oxazolyl, thiazolyl, imidazolyl,
oxadiazolyl, thiadiazolyl, triazolyl, pyridinyl, pyrazinyl,
pyridazinyl, pyrimidinyl, triazinyl, quinolinyl, isoquinolinyl,
tetrahydroisoquinolinyl, quinoxalinyl, quinazolinyl,
naphthyridinyl, indazolyl, indolyl, 2-indolonyl, benzisoxazolyl,
benzoisothiazolyl, benzoxazolyl, benzothiazolyl, benzimidazolyl,
furopyridinyl, thienopyridinyl, thienopyrimidinyl,
isothiazolopyridinyl, thiazolopyridinyl, thiazolopyridinonyl,
thiazolopyrazinyl, thiazolopyrimidinyl, triazolopyridinyl,
triazolopyrazinyl, pyrrolotriazinyl,
5,6-dihydrobenzo[h]quinazolinyl, 5H-chromeno[4,3-d]pyrimidinyl,
6,7-dihydro-5H-cyclopenta[d]pyrimidinyl,
5,6,7,8-tetrahydroquinazolinyl, 7,8-dihydroquinazolin-5(6H)-onyl,
and tetrahydrobenzothiazolyl, and is substituted with 0-3
substituents independently selected from the group consisting of
C.sub.1-4alkyl, C.sub.3-7cycloalkyl, C.sub.1-4haloalkyl,
C.sub.1-4alkoxy, C.sub.1-4haloalkoxy, C.sub.3-7cycloalkoxy,
C.sub.1-4alkylthio, phenoxy, benzyloxy, halo, hydroxy, cyano,
nitro, C.sub.1-4alkylsulfonyl, NR.sup.2R.sup.3, pyrrolidinonyl,
methylenedioxy, furyl, thienyl, pyrazolyl, imidazolyl, thiazolyl,
triazolyl, pyrazinyl, pyrimidinyl, naphthyl, C.sub.1-4alkylamido,
CONR.sup.2R.sup.3, pyridyl, phenyl, and benzyl, and where
imidazolyl, pyridyl, phenyl and benzyl are substituted with 0-2
substituents independently selected from the group consisting of
halo, C.sub.1-4alkyl, C.sub.1-4alkoxy, C.sub.1-4haloalkyl,
C.sub.1-4haloalkoxy, and NR.sup.2R.sup.3; R.sup.2 is hydrogen,
C.sub.1-4alkyl, C.sub.1-4hydroxyalkyl, or C.sub.1-4-aminoalkyl;
R.sup.3 is hydrogen, C.sub.1-4alkyl, C.sub.1-4hydroxyalkyl, or
C.sub.1-4-aminoalkyl; or R.sub.2 and R.sub.3 taken together with
the nitrogen atom to which they are attached is azetidinyl,
pyrrolidinyl, piperidinyl, piperazinyl,
N--(C.sub.1-4alkyl)piperazinyl, morpholinyl, or homopiperidinyl; or
a pharmaceutically acceptable salt thereof.
[0019] Another aspect of the invention is stereoisomer of formula I
according to formula Ia.
##STR00003##
[0020] Another aspect of the invention is a compound of formula I
or Ia where R.sup.1 is selected from the group consisting of
dimethylisoxazolyl, (methyl)(phenyl)isoxazolyl, methylpyrazolyl,
dimethylpyrazolyl, thienylpyrazolyl, methoxyphenylpyrazolyl,
thiazolyl, bromothiazolyl, cyanothiazolyl, methylthiazolyl,
dimethylthiazolyl, (methyl)(phenyl)thiazolyl, isopropylthiazolyl,
butylthiazolyl, benzylthiazolyl, methoxyphenylmethylthiazolyl,
phenylthiazolyl, chlorophenylthiazolyl, methoxyphenylthiazolyl,
(methoxyphenyl)(methyl)thiazolyl, pyridinylthiazolyl,
(phenyl)(methyl)imidazolyl, methyloxadiazolyl, ethyloxadiazolyl,
methylthiadiazolyl, fluorophenylthiadiazolyl, furylthiadiazolyl,
(dimethylcarboxamido)(methyl)thiazolyl, (pyrrolidinylCO)thiazolyl,
phenyltriazolyl, pyridinyl, bromopyridinyl, chloropyridinyl,
(chloro)(fluoro)pyridinyl, (chloro)(methyl)pyridinyl,
dichloropyridinyl, fluoropyridinyl, cyanopyridinyl,
(cyano)(methyl)pyridinyl, (cyano)(dimethyl)pyridinyl,
methoxypyridinyl, (methylpyrrolidinyl)pyridinyl, phenylpyridinyl,
methoxypyridinylpyridinyl, pyridazinyl, bromopyridazinyl,
chloropyridazinyl, methylpyridazinyl, methoxypyridazinyl,
methylthiopyridazinyl, pyrrolidinylpyridazinyl,
pyrrolidinonylpyridazinyl, phenylpyridazinyl, pyridinylpyridazinyl,
methoxypyridinylpyridazinyl, pyrimidinyl,
(bromo)(isopropyl)pyrimidinyl, (bromo)(dimethyl)pyrimidinyl,
(bromo)(cyclopropyl)pyrimidinyl, (bromo)(methoxy)pyrimidinyl,
(bromo)(phenyl)pyrimidinyl, (bromo)(pyridinyl)pyrimidinyl,
chloropyrimidinyl, (chloro)(dimethyl)pyrimidinyl,
(methyl)(methoxy)pyrimidinyl, methylpyrimidinyl, ethylpyrimidinyl,
(methyl)(phenyl)pyrimidinyl, dimethylpyrimidinyl, butylpyrimidinyl,
isopropylpyrimidinyl, cyclopropylpyrimidinyl, methoxypyrimidinyl,
dimethoxypyrimidinyl, isopropoxypyrimidinyl,
cyclopentoxypyrimidinyl, difluoromethoxypyrimidinyl,
trifluoroethoxypyrimidinyl, phenoxypyrimidinyl,
methylthiopyrimidinyl, phenylpyrimidinyl, chlorophenylpyrimidinyl,
methylphenylpyrimidinyl, methoxyphenylpyrimidinyl,
(phenyl)(triazolyl)pyrimidinyl, pyridinylpyrimidinyl,
methoxypyridinylpyrimidinyl, methoxypyrimidinylpyrimidinyl,
naphthylpyrimidinyl, pyrazinyl, bromopyrazinyl,
(bromo)(methoxy)pyrazinyl, chloropyrazinyl, methylpyrazinyl,
dimethylpyrazinyl, butylpyrazinyl, cyanopyrazinyl,
methoxypyrazinyl, isopropoxypyrazinyl, trifluoromethylpyrazinyl,
and phenylpyrazinyl, and dimethyltriazinyl; or a pharmaceutically
acceptable salt thereof.
[0021] Another aspect of the invention is a compound of formula I
or Ia where R.sup.1 is selected from the group consisting of
dimethylpyridinoisoxazolyl, benzoxazolyl, chlorobenzoxazolyl,
fluorophenylbenzoxazolyl, ethylphenylbenzoxazolyl,
dimethylaminophenylbenzoxazolyl, pyridinylbenzoxazolyl,
benzothiazolyl, acetamidobenzothiazolyl, bromobenzothiazolyl,
chlorobenzothiazolyl, (chloro)(methyl)benzothiazolyl,
(chloro)(methoxy)benzothiazolyl, fluorobenzothiazolyl,
difluorobenzothiazolyl, cyanobenzothiazolyl, methylbenzothiazolyl,
dimethylbenzothiazolyl, (methyl)(methoxy)benzothiazolyl,
ethylbenzothiazolyl, trifluoromethylbenzothiazolyl,
hydroxybenzothiazolyl, methoxybenzothiazolyl, ethoxybenzothiazolyl,
isopropoxybenzothiazolyl, trifluoromethoxybenzothiazolyl,
difluoromethoxybenzothiazolyl, dimethoxybenzothiazolyl,
morpholinylbenzothiazolyl, (pyrrolidinylCO)benzothiazolyl,
methylsulfonylbenzothiazolyl, chlorothiazolopyridinyl,
dimethylthiazolopyridinyl, benzyloxythiazolopyridinyl,
difluoromethoxythiazolopyridinyl, benzotriazolyl, indolonyl,
indazolyl, bromoindazolyl, chloroindazolyl, fluoroindazolyl,
(methyl)(methoxy)indazolyl, methoxyindazolyl,
trifluoromethylindazolyl, trifluoromethoxyindazolyl,
difluoromethoxyindazolyl, benzimidazolyl, fluorobenzimidazolyl,
methylbenzimidazolyl, (methyl)(methoxy)benzimidazolyl,
methoxybenzimidazolyl, tetrahydrobenzothiazolyl, furopyridinyl,
dimethylfuropyrimidinyl, thienopyrimidinyl,
isopropylthienopyrimidinyl, dimethylthienopyrimidinyl,
chlorotriazolopyridinyl, methyltriazolopyridinyl,
trifluoromethyltriazolopyridinyl, methoxytriazolopyridinyl,
triazolopyrazinyl, bromopyrrolotriazinyl,
dimethylaminothiazolopyrimidinyl, thiazolopyazinyl,
bromothiazolopyazinyl, methoxythiazolopyazinyl,
methylthiothiazolopyazinyl, methoxythiazolopyrimidinyl,
(methyl)(methoxy)thiazolopyrimidinyl, quinolinyl, bromoquinolinyl,
fluoroquinolinyl, methylquinolinyl, (methyl)(methoxy)quinolinyl,
isoquinolinyl, bromoisoquinolinyl, dichloroisoquinolinyl,
methylisoquinolinyl, dimethylisoquinolinyl, quinoxalinyl,
chloroquinoxalinyl, methylquinoxalinyl, methoxyquinoxalinyl,
quinazolinyl, bromoquinazolinyl, naphthyridinyl,
5,6-dihydrobenzo[h]quinazolinyl, 5H-chromeno[4,3-d]pyrimidinyl,
6,7-dihydro-5H-cyclopenta[d]pyrimidinyl,
5,6,7,8-tetrahydroquinazolinyl, and
7,8-dihydroquinazolin-5(6H)-onyl; or a pharmaceutically acceptable
salt thereof.
[0022] Another aspect of the invention is a compound of formula I
or Ia where R.sup.1 is selected from the group consisting of
phenylthiazolyl, (chloro)(methyl)pyridinyl,
(bromo)(phenyl)pyrimidinyl, methoxypyrimidinyl,
difluoromethoxypyrimidinyl, difluoroethoxypyrimidinyl,
cyclopentoxypyrimidinyl, (methylphenyl)pyrimidinyl,
(methoxyphenyl)pyrimidinyl, bromopyrazinyl, chloropyrazinyl,
methylthiopyrazinyl, methoxybenzothiazolyl, ethoxybenzothiazolyl,
difluoromethoxybenzothiazolyl, thiazolopyridinonyl,
trifluoromethylindazolyl, benzimidazolyl, isoquinoinyl, and
quinazolinyl or a pharmaceutically acceptable salt thereof.
[0023] Another aspect of the invention is a compound or formula I
or Ia where R.sup.1 is selected from the group consisting of
bromopyridinyl, dichloropyridinyl, methoxypyridinyl,
(pyridinyl)pyridinyl, (phenyl)pyrimidinyl,
(methoxypyridinyl)pyrimidinyl, (pyrazolyl)pyrimidinyl,
chloropyrazinyl, (bromo)(chloro)pyrazinyl, and
chlorobenzothiazolyl; or a pharmaceutically acceptable salt
thereof.
[0024] Another aspect of the invention is a compound or formula I
or Ia where R.sup.1 is selected from the group consisting of
thiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl,
benzothiazolyl, thiazolopyridinyl, indazolyl, benzimidazolyl,
isoquinolinyl, and quinazolinyl, and is substituted with 0-3
substituents independently selected from the group consisting of
C.sub.1-4alkyl, C.sub.3-7cycloalkyl, C.sub.1-4haloalkyl,
C.sub.1-4alkoxy, C.sub.1-4haloalkoxy, C.sub.3-7cycloalkoxy,
C.sub.1-4alkylthio, phenoxy, benzyloxy, halo, hydroxy, cyano,
C.sub.1-4alkylsulfonyl, NR.sup.2R.sup.3, pyrrolidinonyl,
methylenedioxy, furyl, thienyl, triazolyl, pyrimidinyl, naphthyl,
C.sub.1-4alkylamido, CONR.sup.2R.sup.3, pyridyl, phenyl, and
benzyl, and where pyridyl, phenyl and benzyl are substituted with
0-2 substituents independently selected from the group consisting
of halo, C.sub.1-4alkyl, C.sub.1-4alkoxy, C.sub.1-4haloalkyl,
C.sub.1-4haloalkoxy, and NR.sup.2R.sup.3; or a pharmaceutically
acceptable salt thereof.
[0025] Another aspect of the invention is a compound or formula I
or Ia where R.sup.1 is selected from the group consisting of
pyridinyl, pyrimidinyl, pyrazinyl, thiazolopyridinyl, and
isoquinolinyl, and is substituted with 0-3 substituents
independently selected from the group consisting of C.sub.1-4alkyl,
C.sub.3-7cycloalkyl, C.sub.1-4haloalkyl, C.sub.1-4alkoxy,
C.sub.1-4haloalkoxy, C.sub.3-7cycloalkoxy, C.sub.1-4alkylthio,
phenoxy, benzyloxy, halo, hydroxy, cyano, C.sub.1-4alkylsulfonyl,
NR.sup.2R.sup.3, pyrrolidinonyl, methylenedioxy, furyl, thienyl,
triazolyl, pyrimidinyl, naphthyl, C.sub.1-4alkylamido,
CONR.sup.2R.sup.3, pyridyl, phenyl, and benzyl, and where pyridyl,
phenyl and benzyl are substituted with 0-2 substituents
independently selected from the group consisting of halo,
C.sub.1-4alkyl, C.sub.1-4alkoxy, C.sub.1-4haloalkyl,
C.sub.1-4haloalkoxy, and NR.sup.2R.sup.3; or a pharmaceutically
acceptable salt thereof.
[0026] Another aspect of the invention is a compound or formula I
or Ia where R.sup.1 is selected from the group consisting of
pyridinyl and isoquinolinyl, and is substituted with 0-3
substituents independently selected from the group consisting of
C.sub.1-4alkyl, C.sub.3-7cycloalkyl, C.sub.1-4haloalkyl,
C.sub.1-4alkoxy, C.sub.1-4haloalkoxy, C.sub.3-7cycloalkoxy,
C.sub.1-4alkylthio, phenoxy, benzyloxy, halo, hydroxy, cyano,
C.sub.1-4alkylsulfonyl, NR.sup.2R.sup.3, pyrrolidinonyl,
methylenedioxy, furyl, thienyl, triazolyl, pyrimidinyl, naphthyl,
C.sub.1-4alkylamido, CONR.sup.2R.sup.3, pyridyl, phenyl, and
benzyl, and where pyridyl, phenyl and benzyl are substituted with
0-2 substituents independently selected from the group consisting
of halo, C.sub.1-4alkyl, C.sub.1-4alkoxy, C.sub.1-4haloalkyl,
C.sub.1-4haloalkoxy, and NR.sup.2R.sup.3; or a pharmaceutically
acceptable salt thereof.
[0027] Another aspect of the invention is a compound selected from
the group consisting of
##STR00004##
or a pharmaceutically acceptable salt thereof.
[0028] Another aspect of the invention is a compound selected from
the group consisting of
##STR00005## ##STR00006##
or a pharmaceutically acceptable salt thereof.
[0029] Another aspect of the invention is a compound of formula I
where R.sub.1 is selected from the group consisting of thiazole,
thiadiazole, isoxazole, oxazole, pyrazole, imidazole, pyridine,
pyrazine, pyridazine, pyrimidine, quinoline, isoquinoline,
quinoxaline, indazole, indole, 2-indolone, benzothiazole,
benzimidazole, benzoxazole, benzo[d]isothiazole, benzisoxazole,
isothiazolo-[5,4-b]pyridine, (1,2,4)-triazolo[1,5-a]pyridine,
thiazolo[5,4-b]pyridine and tetrahydrobenzothiazole in which each
group is optionally substituted with one or two substituents
selected from the group consisting of C.sub.1-4alkyl,
C.sub.1-4alkoxy, halogen, hydroxy, cyano, trifluoromethyl,
difluoromethyl, fluoromethyl, trifluoromethoxy, difluoromethoxy,
C.sub.1-4alkylsulfonyl, furyl, morpholino, methylenedioxy, pyridyl,
C.sub.1-4alkylphenyl, halophenyl, dimethylaminophenyl,
C.sub.1-4alkylamido, --CONR.sub.2R.sub.3 in which R.sub.2 and
R.sub.3 each are independently hydrogen, C.sub.1-4alkyl, hydroxy
C.sub.1-4alkyl, amino C.sub.1-4alkyl or R.sub.2 and R.sub.3 taken
together with the atom to which they are attached are C.sub.3-6
cycloalkyl; phenyl, substituted phenyl, phenylmethyl, substituted
phenylmethyl in which said substituted phenyl and substituted
phenylmethyl are substituted with substituents independently
selected from the group consisting of halogen, C.sub.1-4alkyl,
C.sub.1-4alkoxy, trifluoromethyl and trifluoromethoxy; or a
pharmaceutically acceptable salt thereof.
[0030] For a compound of formula I or Ia, the scope of any instance
of a variable substituent, including R.sup.1, R.sup.2, and R.sup.3,
can be used independently with the scope of any other instance of a
variable substituent. As such, the invention includes combinations
of the different aspects.
[0031] Unless specified otherwise, these terms have the following
meanings. "Alkyl" means a straight or branched alkyl group composed
of 1 to 4 carbons. "Alkenyl" means a straight or branched alkyl
group composed of 2 to 4 carbons with at least one double bond.
"Alkynyl" means a straight or branched alkyl group composed of 2 to
4 carbons with at least one triple bond. "Cycloalkyl" means a
monocyclic ring system composed of 3 to 7 carbons. "Haloalkyl" and
"haloalkoxy" include all halogenated isomers from monohalo to
perhalo. Terms with a hydrocarbon moiety (e.g. alkoxy) include
straight and branched isomers for the hydrocarbon portion.
Parenthetic and multiparenthetic terms are intended to clarify
bonding relationships to those skilled in the art. For example, a
term such as ((R)alkyl) means an alkyl substituent further
substituted with the substituent R.
[0032] The invention includes all pharmaceutically acceptable salt
forms of the compounds. Pharmaceutically acceptable salts are those
in which the counter ions do not contribute significantly to the
physiological activity or toxicity of the compounds and as such
function as pharmacological equivalents. These salts can be made
according to common organic techniques employing commercially
available reagents. Some anionic salt forms include acetate,
acistrate, besylate, bromide, chloride, citrate, fumarate,
glucouronate, hydrobromide, hydrochloride, hydroiodide, iodide,
lactate, maleate, mesylate, nitrate, pamoate, phosphate, succinate,
sulfate, tartrate, tosylate, and xinofoate. Some cationic salt
forms include ammonium, aluminum, benzathine, bismuth, calcium,
choline, diethylamine, diethanolamine, lithium, magnesium,
meglumine, 4-phenylcyclohexylamine, piperazine, potassium, sodium,
tromethamine, and zinc.
[0033] Some of the compounds of the invention exist in
stereoisomeric forms. The invention includes all stereoisomeric
forms of the compounds including enantiomers and diastereromers.
Methods of making and separating stereoisomers are known in the
art.
[0034] The invention includes all tautomeric forms of the
compounds. An example of a tautomeric pair is shown below.
##STR00007##
[0035] The invention is intended to include all isotopes of atoms
occurring in the present compounds. Isotopes include those atoms
having the same atomic number but different mass numbers. By way of
general example and without limitation, isotopes of hydrogen
include deuterium and tritium. Isotopes of carbon include .sup.13C
and .sup.14C. Isotopically-labeled compounds of the invention can
generally be prepared by conventional techniques known to those
skilled in the art or by processes analogous to those described
herein, using an appropriate isotopically-labeled reagent in place
of the non-labeled reagent otherwise employed. Such compounds may
have a variety of potential uses, for example as standards and
reagents in determining biological activity. In the case of stable
isotopes, such compounds may have the potential to favorably modify
biological, pharmacological, or pharmacokinetic properties.
Synthetic Methods
[0036] The compounds may be made by methods known in the art
including those described below and including variations within the
skill of the art. Some reagents and intermediates are known in the
art. Other reagents and intermediates can be made by methods known
in the art using readily available materials. The variables (e.g.
numbered "R" substituents) used to describe the synthesis of the
compounds are intended only to illustrate how to make the compounds
and are not to be confused with variables used in the claims or in
other sections of the specification. The following methods are for
illustrative purposes and are not intended to limit the scope of
the invention.
[0037] Some of the compounds may be prepared using the reactions
and techniques described in this section. The reactions are
performed in solvents appropriate to the reagents and materials
employed and are suitable for the transformations being effected.
It is understood by one skilled in the art of organic synthesis
that the functionality present on various portions of the molecule
must be compatible with the reagents and reactions proposed. Such
restrictions to the substituents which are compatible with the
reaction conditions will be readily apparent to one skilled in the
art and alternate methods must then be used.
[0038] Abbreviations used in the schemes generally follow
conventions used in the art. Chemical abbreviations used in the
specification and examples are defined as follows: "NaHMDS" for
sodium bis(trimethylsilyl)amide; "DMF" for N,N-dimethylformamide;
"MeOH" for methanol; "NBS" for N-bromosuccinimide; "Ar" for aryl;
"TFA" for trifluoroacetic acid; "LAH" for lithium aluminum hydride;
"BOC", "DMSO" for dimethylsulfoxide; "h" for hours; "rt" for room
temperature or retention time (context will dictate); "min" for
minutes; "EtOAc" for ethyl acetate; "THF" for tetrahydrofuran;
"EDTA" for ethylenediaminetetraacetic acid; "Et.sub.2O" for diethyl
ether; "DMAP" for 4-dimethylaminopyridine; "DCE" for
1,2-dichloroethane; "ACN" for acetonitrile; "DME" for
1,2-dimethoxyethane; "HOBt" for 1-hydroxybenzotriazole hydrate;
"DIEA" for diisopropylethylamine, "Nf" for
CF.sub.3(CF.sub.2).sub.3SO.sub.2--; and "TMOF" for
trimethylorthoformate.
[0039] Abbreviations as used herein, are defined as follows:
"1.times." for once, "2.times." for twice, "3.times." for thrice,
".degree. C." for degrees Celsius, "eq" for equivalent or
equivalents, "g" for gram or grams, "mg" for milligram or
milligrams, "L" for liter or liters, "mL" for milliliter or
milliliters, ".mu.L" for microliter or microliters, "N" for normal,
"M" for molar, "mmol" for millimole or millimoles, "min" for minute
or minutes, "h" for hour or hours, "rt" for room temperature, "RT"
for retention time, "atm" for atmosphere, "psi" for pounds per
square inch, "conc." for concentrate, "sat" or "sat'd" for
saturated, "MW" for molecular weight, "mp" for melting point, "cc"
for enantiomeric excess, "MS" or "Mass Spec" for mass spectrometry,
"ESI" for electrospray ionization mass spectroscopy, "HR" for high
resolution, "HRMS" for high resolution mass spectrometry, "LCMS"
for liquid chromatography mass spectrometry, "HPLC" for high
pressure liquid chromatography, "RP HPLC" for reverse phase HPLC,
"TLC" or "tlc" for thin layer chromatography, "NMR" for nuclear
magnetic resonance spectroscopy, ".sup.1H" for proton, ".delta."
for delta, "s" for singlet, "d" for doublet, "t" for triplet, "q"
for quartet, "m" for multiplet, "br" for broad, "Hz" for hertz, and
".alpha.", ".beta.", "R", "S", "E", and "Z" are stereochemical
designations familiar to one skilled in the art.
##STR00008##
[0040] Compounds of Formula I are prepared as illustrated in
Reaction Scheme 1. The ketone of Formula III (3-quinuclidone) is
known, is commercially available, or may be prepared by methods
known to those skilled in the art. The ketone can be converted to
the corresponding cyanohydrin of Formula IV by reaction with sodium
or potassium cyanide plus an acid. The compound of Formula IV can
be reduced to the corresponding amino-methyl compound (borane
complex) of Formula V by reaction with borane/tetrahydrofuran
complex.
[0041] The compound of Formula V can be reacted with heteroaryl
isothiocyanates directly in an inert solvent to give the thioureas
of Formula VI. Alternatively, the heteroarylamine can be reacted
with thiocarbonyl-diimidazole to give an activated species which
can be used without isolation to convert the compound of Formula V
to the compound of Formula VI. The heteroarylamine may be prepared
by methods known to those skilled in the art.
[0042] The thiourea of Formula VI can be cyclized using, for
example, di-isopropyl carbodiimide to give the oxazoline of Formula
VII which may be deprotected via treatment with acid to give the
racemic final product of the compound of Formula I. The compound of
Formula I may be resolved into pure enantiomer compounds of Formula
Ia and Formula Ib by means known in the art, for example, via
chiral chromatography.
##STR00009##
[0043] Alternatively, the free amino group of the quinuclidine of
Formula V can be blocked with, for example, carbobenzyloxy-chloride
("CBZ--Cl") to give the compound of Formula VIII, as illustrated in
Reaction Scheme 2.
[0044] The racemic compound of Formula VIII can be resolved into
its enantiomers, Formula IX and Formula X by, for example, chiral
chromatography. Either the compound of Formula IX or Formula X, and
preferably the compound of Formula X, can then be carried on as
shown in Reaction Scheme 2.
[0045] The borane group in the compound of Formula X can be
removed, for example, by treatment with dilute hydrochloric acid,
and the carbobenzyloxy group can be removed, for example, by
catalytic hydrogenation to give the chiral quinuclidine amine of
Formula XI. Similarly to Reaction Scheme 1, the amine salt of
Formula XI can be reacted with isothiocyanates to give the thiourea
of Formula VIa, which can then be reacted with dialkyl
carbodiimides or mixed thioureas (as from reaction with
thiocarbonyl diimidazole) to give the chiral oxazoline quinuclidine
compounds of Formula Ia, and its tautomer, Formula IIa.
##STR00010##
[0046] Alternatively, the borane group of V may be removed with
hydrochloric acid to give dihydrochloride salt XII, which can be
reacted in the presence of base with an isothiocyanate to give
intermediate thiourea XIII, which can then be cyclized as in
Reaction Schemes 1-2 to give I. XII may also be prepared by other
methods, as referenced in U.S. Pat. No. 5,137,895 (Aug. 11,
1992).
##STR00011##
[0047] Additionally, the (hetero)aromatic amines may be reacted
with carbon disulfide, sodium hydroxide, and methyl iodide to give
intermediate dimethyl carbonimidodithioates XIV. These are reacted
with dihydrochloride XI in the presence of base to eliminate two
moles of methanethiol and generate desired products Ia
directly.
Biological Methods
[0048] I) .alpha.7 Nicotinic Acetycholine Receptor Binding.
Membranes were prepared for binding using HEK293 cells stably
expressing the rat .alpha.7 nicotinic acetycholine receptor (rat
.alpha.7 nAChR). Cells were homogenized at 4.degree. C. in
hypotonic lysis buffer consisting of 10 mM Tris (pH 7.4), 5 mM EDTA
and protease inhibitors and centrifuged at 32000.times.g for 20
minutes. The pellet was washed once in membrane wash buffer
consisting of 50 mM Tris (pH 7.4), 1 mM EDTA and protease
inhibitors and centrifuged at 32000.times.g for 20 minutes. This
pellet was then resuspended in assay buffer consisting 50 mM
KH.sub.2PO.sub.4 (pH 7.4 at 25.degree. C.), 1 mM EDTA, 0.005%
Triton-X 100 and 0.1% (v/v) Sigma Protease Inhibitor Cocktail.
Aliquots were then frozen in dry ice/ethanol and kept at
-80.degree. C. until the day of the assay. II) A
Ca.sup.2+-Sensitive, Fluorescence-Based Assay .alpha.-7 for
Nicotinic Acetylcholine Receptor Channel Function In Mammalian
Cells ("FLIPR"). Summary: Lead compounds are evaluated for agonist
activity at .alpha.-7, .alpha.3.beta.4, .alpha.4.alpha..beta.2, and
.alpha.1.beta.1.delta.1.epsilon. sub-types of nicotinic ACh
receptor ion channels expressed in mammalian HEK 293 cells. Agonist
potency and efficacy values are determined from kinetic
fluorescence Ca.sup.2+ influx measurements made using a 384 well
FLIPR (Fluorescence Image Plate Reader). The utility of fluorescent
indicators for measuring changes in intracellular divalent cation
concentrations, particularly Ca.sup.2+, for drug discovery
endeavors is well documented (Rudiger, R., et al., Nature Reviews,
2003, 4:579-586; Gonzalez J. E., et al., Receptors and Channels,
2002, 8:283-295). In this assay, channel expressing HEK cell lines
seeded in 384 well assay plates are loaded with a membrane permeant
fluorescent Ca.sup.2+ indicator dye, whose 510 nm green emission
signal increases in response to elevation of intracellular
Ca.sup.2+ concentration. The basal fluorescence from the cells is
monitored in real time, followed by the acute addition of test
compounds. If the compound is an agonist at any of the
non-selective cation channels, the latter open and allow the
movement of extracellular Ca.sup.2+ ions into the cell cytoplasm,
where they bind to the Ca.sup.2+ indicator dye, and produce an
increase in fluorescence output signal, which is detected by a
cooled CCD imaging camera.
[0049] Materials and Methods: Reagents: The acetomethoxy (AM) ester
of the Ca.sup.2+ indicator dye Fluo-4 was obtained from InVitrogen,
(Carlsbad, Calif.). Acetylcholine and all buffer constituents were
purchased from Sigma Chemical Company, St. Louis, Mo. G418 and
Minimal Essential Medium were purchased from InVitrogen Life
Technologies, Carlsbad, Calif. Fetal bovine serum was purchased
from (InVitrogen, Carlsbad, Calif.).
[0050] Cell Culture: HEK-293 cells were grown in Minimal Essential
Medium containing 10% (v/v) fetal bovine serum at 37.degree. C. in
a 5% CO.sub.2 incubator. HEK-293 cells stably expressing the ion
channels were grown in the same medium with the addition of 500
.mu.g/ml G418.
[0051] Ca.sup.2+ flux assays of Ca.sup.2+ channels expressed in
HEK-293 cells: HEK-293 cells expressing the ion channels of
interest were plated in 384 well, black-walled, clear-bottomed,
poly-D-lysine coated plates at a density of .about.20,000
cells/well in 20 .mu.l of Minimal Essential Medium containing 10%
(v/v) fetal bovine serum and incubated for 2 days at 29.degree. C.
in a 5% CO.sub.2 incubator. Prior to assay, cells were loaded with
the Fluo-4 AM ester. Cell loading was accomplished by removing the
culture medium and replacing it with 30 .mu.l/well of the AM ester
of the dye (5 .mu.M) mixed with Hanks Balanced Salt Solution
(#14175-095) containing 20 mM HEPES, 2.5 mM probenecid, 0.5 mM
CaCl.sub.2, 1 mM MgCl2 and 10 .mu.M atropine. Dye loading was
allowed to proceed for 90 minutes at room temperature at which time
the dye loading solution was removed and replaced with 40
.mu.l/well of Hanks buffer. Cells loaded with dye were loaded onto
a FLIPR384 (Molecular Devices, Sunnyvale, Calif.). Fluo-4 dye was
excited using the 488 nm line of an argon laser. Emission was
filtered using a 540+/-30 nm bandpass filter. For evaluation of the
effects of test compounds using the Ca.sup.2+ flux assay, compounds
to be tested were provided in assay ready plates. For nicotinic
receptor ion channel expressing cells, the assay was initiated by
the addition of 20 .mu.l/well of Hanks buffer containing test
compounds. For all assays, data were collected at 1 Hz for 10
seconds (baseline), at which time the compound containing stimulus
buffers are added, and further measurements collected at 0.33 Hz
for 3 min.
[0052] Data Analysis: The statistical robustness of the nicotinic
receptor Ca.sup.2+ flux assays is determined from blanks and totals
wells. The totals wells define maximal channel activation for each
compound test plate (Maximum efficacious dose of acetylcholine),
and the blanks wells which contain matched DMSO only, define zero
channel activation. The raw fluorescence units data files generated
on the FLIPR plate reader are automatically exported and processed
by in-house data analysis tools. The reduced percent activation
data for each concentration of test compound are fit using MathIQ
fitting engine (ID Business Solutions Limited, Surrey, UK). Data
were analyzed by fitting maximum amplitudes of change in
fluorescence, for Ca.sup.2+ flux for a given condition of test
compound. Potencies (EC.sub.50 values) of compounds are calculated
from the average of three assay wells from a twenty point CRC. Test
compound efficacy values (Ymax values) are expressed relative to a
maximal response to acetylcholine in the total wells.
III) Fos quantification assay: Male Wistar rats are treated with
drug (1-10 mg/kg) or vehicle (2 ml/kg, sc). Two hours after
treatments, the rats are rapidly decapitated and discrete brain
regions of interest are isolated on ice and weighed and flash
frozen with liquid nitrogen and stored at -80 deg. C. Further
processing of the brain tissue for nuclear extracts as well as for
Fos quantification are in accordance with the protocol prescribed
by a commercially available ELISA-based chemiluminiscence detection
kit (catalog #89860, EZ-detect c-Fos Trans kit, Pierce
Biotechnology Inc., Rockford, Ill.). IV) MK-801 Disrupted Set-Shift
Assay in rats: This assay uses a modification of the protocol
described by Stefani et al. (Behavioral Neuroscience, 2003, 117:
728-737). Test compounds are assessed for their ability to reverse
an MK-801-induced performance deficit (0.03 mg/kg, i.p., single
dose) in this assay.
[0053] The activity of specific compounds described herein and
tested in the above assay (II) is provided in Table 1.
TABLE-US-00001 TABLE 1 ##STR00012## FLIPR Ex .alpha.7 amp- FLIPR
activity.sup.a le .alpha.7 rating Num- (EC.sub.50, (EC.sub.50, ber
R.sub.1 nM) nM) 1 ##STR00013## +++ 1a ##STR00014## + 1b
##STR00015## +++ 2 ##STR00016## ++ 2a ##STR00017## + 2b
##STR00018## ++ 3 ##STR00019## 3840 + 4 ##STR00020## +++ 4a
##STR00021## + 4b ##STR00022## 230 ++ 5 ##STR00023## ++ 5a
##STR00024## + 5b ##STR00025## +++ 6 ##STR00026## ++ 6a
##STR00027## 4340 + 6b ##STR00028## +++ 7 ##STR00029## ++ 7a
##STR00030## + 7b ##STR00031## +++ 8 ##STR00032## ++ 8a
##STR00033## 120 ++ 8b ##STR00034## + 9 ##STR00035## ++ 9a
##STR00036## 7315 + 9b ##STR00037## +++ 10 ##STR00038## ++ 10a
##STR00039## ++ 10b ##STR00040## ++ 11 ##STR00041## ++ 11b
##STR00042## 213 ++ 12 ##STR00043## ++ 12a ##STR00044## ++ 12b
##STR00045## + 13 ##STR00046## +++ 13a ##STR00047## ++ 13b
##STR00048## +++ 14 ##STR00049## ++ 14b ##STR00050## ++ 15
##STR00051## ++ 15a ##STR00052## 6345 + 15b ##STR00053## ++ 16
##STR00054## ++ 16a ##STR00055## 3174 + 16b ##STR00056## ++ 17
##STR00057## ++ 18 ##STR00058## 3930 + 19 ##STR00059## ++ 20
##STR00060## ++ 20a ##STR00061## ++ 20b ##STR00062## 318 ++ 21
##STR00063## +++ 22 ##STR00064## ++ 23 ##STR00065## +++ 24
##STR00066## +++ 25 ##STR00067## 510 ++ 26 ##STR00068## +++ 27
##STR00069## +++ 28 ##STR00070## 4010 + 29 ##STR00071## ++ 30
##STR00072## ++ 31 ##STR00073## ++ 32 ##STR00074## ++ 34
##STR00075## + 35 ##STR00076## 4035 + 36 ##STR00077## ++ 37
##STR00078## + 38 ##STR00079## +++ 39 ##STR00080## +++ 40
##STR00081## +++ 41 ##STR00082## +++ 42 ##STR00083## +++ 43
##STR00084## +++ 44 ##STR00085## ++ 45 ##STR00086## ++ 46
##STR00087## 2000 + 47 ##STR00088## +++ 48 ##STR00089## 790 ++ 49
##STR00090## 3330 + 50 ##STR00091## +++ 51 ##STR00092## NT.sup.b 52
##STR00093## 325 ++ 53 ##STR00094## + 54 ##STR00095## + 55
##STR00096## + 56 ##STR00097## + 57 ##STR00098## 3900 + 58
##STR00099## + 59 ##STR00100## 4240 + 60 ##STR00101## 2550 + 61
##STR00102## ++ 62 ##STR00103## + 63 ##STR00104## 2600 + 64
##STR00105## +++ 65 ##STR00106## + 66 ##STR00107## 360 ++ 67
##STR00108## + 68 ##STR00109## + 70 ##STR00110## ++ 71 ##STR00111##
+ 72 ##STR00112## + 73 ##STR00113## 7550 + 74 ##STR00114## +++ 75
##STR00115## + 76 ##STR00116## ++ 77 ##STR00117## 355 ++ 78
##STR00118## + 81 ##STR00119## NT.sup.b 82 ##STR00120## NT.sup.b 83
##STR00121## 240 ++ 85 ##STR00122## + 86 ##STR00123## + 87
##STR00124## +++ 88 ##STR00125## ++ 89 ##STR00126## +++ 90
##STR00127## 130 ++ 91 ##STR00128## ++ 92 ##STR00129## +++ 93
##STR00130## ++ 94 ##STR00131## +++ 95 ##STR00132## ++
96 ##STR00133## + 97 ##STR00134## ++ 98 ##STR00135## + 99
##STR00136## ++ 100 ##STR00137## +++ 101 ##STR00138## ++ 102
##STR00139## +++ 103 ##STR00140## 280 ++ 104 ##STR00141## ++ 105
##STR00142## ++ 106 ##STR00143## ++ 107 ##STR00144## ++ 108
##STR00145## NT.sup.b 109 ##STR00146## ++ 110 ##STR00147## 450 ++
111 ##STR00148## ++ 112 ##STR00149## +++ 113 ##STR00150## +++ 114
##STR00151## ++ 115 ##STR00152## + 116 ##STR00153## + 117
##STR00154## ++ 118 ##STR00155## + 119 ##STR00156## +++ 120
##STR00157## 875 ++ 121 ##STR00158## + 122 ##STR00159## +++ 123
##STR00160## + 124 ##STR00161## + 125 ##STR00162## 17 +++ 126
##STR00163## +++ 127 ##STR00164## + 128 ##STR00165## +++ 129
##STR00166## ++ 130 ##STR00167## ++ 131 ##STR00168## + 132
##STR00169## ++ 133 ##STR00170## +++ 134 ##STR00171## +++ 135
##STR00172## +++ 136 ##STR00173## +++ 137 ##STR00174## 137 ++ 138
##STR00175## + 139 ##STR00176## 16 +++ 140 ##STR00177## ++ 141
##STR00178## 4 +++ 142 ##STR00179## ++ 143 ##STR00180## ++ 144
##STR00181## +++ 145 ##STR00182## + 146 ##STR00183## ++ 147
##STR00184## ++ 148 ##STR00185## + 149 ##STR00186## 17 +++ 150
##STR00187## ++ 151 ##STR00188## ++ 152 ##STR00189## 233 ++ 153
##STR00190## ++ 154 ##STR00191## + 155 ##STR00192## ++ 156
##STR00193## ++ 157 ##STR00194## ++ 158 ##STR00195## 135 ++ 159
##STR00196## +++ 160 ##STR00197## + 161 ##STR00198## + 162
##STR00199## + 163 ##STR00200## +++ 164 ##STR00201## ++ 165
##STR00202## 13 +++ 166 ##STR00203## 11 +++ 167 ##STR00204## + 168
##STR00205## +++ 169 ##STR00206## + 170 ##STR00207## +++ 171
##STR00208## +++ 172 ##STR00209## ++ 173 ##STR00210## ++ 174
##STR00211## 245 ++ 175 ##STR00212## ++ 176 ##STR00213## ++ 177
##STR00214## +++ 178 ##STR00215## 17 +++ 179 ##STR00216## ++ 180
##STR00217## ++ 181 ##STR00218## 13 +++ 182 ##STR00219## ++ 183
##STR00220## +++ 184 ##STR00221## + 185 ##STR00222## + 186
##STR00223## ++ 187 ##STR00224## 424 ++ 188 ##STR00225## + 189
##STR00226## ++ 190 ##STR00227## + 191 ##STR00228## +++ 192
##STR00229## ++ 193 ##STR00230## 17 +++ 194 ##STR00231## 18 +++ 195
##STR00232## + 196 ##STR00233## 13 +++ 197 ##STR00234## +++ 198
##STR00235## 15 +++ 199 ##STR00236## + 200 ##STR00237## +++ 201
##STR00238## +++ 202 ##STR00239## 16 +++ 203 ##STR00240## +++ 204
##STR00241## ++ 205 ##STR00242## ++ 206 ##STR00243## + 207
##STR00244## ++ 208 ##STR00245## + 209 ##STR00246## +++ 210
##STR00247## 9 +++ 211 ##STR00248## ++ 212 ##STR00249## + 213
##STR00250## 9 +++ 214 ##STR00251## +++ 215 ##STR00252## +++ 216
##STR00253## + 217 ##STR00254## + 218 ##STR00255## 285 ++ 219
##STR00256## + 220 ##STR00257## +++
221 ##STR00258## ++ 222 ##STR00259## ++ 223 ##STR00260## ++ 224
##STR00261## ++ 225 ##STR00262## ++ 226 ##STR00263## +++ 227
##STR00264## ++ 228 ##STR00265## +++ 229 ##STR00266## +++ 230
##STR00267## +++ 231 ##STR00268## ++ 232 ##STR00269## ++ 233
##STR00270## +++ 234 ##STR00271## +++ 235 ##STR00272## + 236
##STR00273## + 237 ##STR00274## +++ 238 ##STR00275## 16 +++ 239
##STR00276## 15 +++ 240 ##STR00277## ++ 241 ##STR00278## + 242
##STR00279## ++ 243 ##STR00280## + 244 ##STR00281## +++ 245
##STR00282## +++ 246 ##STR00283## +++ 247 ##STR00284## ++ 248
##STR00285## ++ 249 ##STR00286## + 250 ##STR00287## +++ 251
##STR00288## +++ 252 ##STR00289## +++ 253 ##STR00290## +++ 254
##STR00291## ++ 255 ##STR00292## ++ 256 ##STR00293## +++ 257
##STR00294## +++ 258 ##STR00295## + 259 ##STR00296## + 260
##STR00297## ++ 261 ##STR00298## +++ 262 ##STR00299## + 263
##STR00300## +++ 264 ##STR00301## ++ 265 ##STR00302## +++ 266
##STR00303## + 267 ##STR00304## +++ 268 ##STR00305## +++ 269
##STR00306## + 270 ##STR00307## +++ 271 ##STR00308## +++ 272
##STR00309## +++ 273 ##STR00310## + 274 ##STR00311## + 275
##STR00312## ++ 276 ##STR00313## +++ 277 ##STR00314## +++ 278
##STR00315## +++ 279 ##STR00316## ++ 280 ##STR00317## 281
##STR00318## +++ 282 ##STR00319## ++ 283 ##STR00320## ++ 284
##STR00321## +++ 285 ##STR00322## ++ 286 ##STR00323## ++ 287
##STR00324## ++ 288 ##STR00325## +++ 289 ##STR00326## +++ 290
##STR00327## +++ 291 ##STR00328## +++ 292 ##STR00329## +++ 293
##STR00330## ++ 294 ##STR00331## +++ 295 ##STR00332## ++
.sup.aActivity based on EC.sub.50 nM values: +++ = <100 nM; ++ =
100-1000 nM; + = 1000-100000 nM; .sup.bNT = Not tested; NA = Not
active (>1000000 nM).
TABLE-US-00002 TABLE Ib ##STR00333## FLIPR .alpha.7 activity.sup.a
FLIPR .alpha.7 rating Example (EC.sub.50, (EC.sub.50, Number
R.sub.1 nM) nM) 296 ##STR00334## +++ 297 ##STR00335## 35 +++ 298
##STR00336## 25 +++ 299 ##STR00337## +++ 300 ##STR00338## +++ 301
##STR00339## +++ 302 ##STR00340## 9 +++ 303 ##STR00341## +++ 304
##STR00342## +++ 305 ##STR00343## +++ 306 ##STR00344## +++ 307
##STR00345## +++ 308 ##STR00346## 59 +++ 309 ##STR00347## +++ 310
##STR00348## +++ 311 ##STR00349## +++ 312 ##STR00350## +++ 313
##STR00351## +++ 314 ##STR00352## +++ 315 ##STR00353## +++ 316
##STR00354## +++ 317 ##STR00355## 660 ++ 318 ##STR00356## +++ 319
##STR00357## +++ 320 ##STR00358## +++ 321 ##STR00359## +++ 322
##STR00360## +++ 323 ##STR00361## +++ 324 ##STR00362## +++ 325
##STR00363## 139 ++ 326 ##STR00364## ++ 327 ##STR00365## ++ 328
##STR00366## ++ 329 ##STR00367## +++ 330 ##STR00368## +++ 331
##STR00369## +++ 332 ##STR00370## +++ 333 ##STR00371## +++ 334
##STR00372## +++ 335 ##STR00373## 104 ++ 336 ##STR00374## +++ 337
##STR00375## ++ 338A ##STR00376## +++ 338B ##STR00377## +++ 339
##STR00378## +++ 340 ##STR00379## +++ 341 ##STR00380## +++ 342
##STR00381## 1600 + 343 ##STR00382## +++ 344 ##STR00383## +++ 345
##STR00384## +++ 346 ##STR00385## 310 ++ 347 ##STR00386## +++ 348
##STR00387## +++ 349 ##STR00388## +++ 350 ##STR00389## NT 351
##STR00390## +++ 352 ##STR00391## 560 ++ S-Enantiomer
.sup.aActivity based on EC.sub.50 nM values: +++ = <100 nM; ++ =
100-1000 nM; + = 1000-100000 nM; .sup.bNT = Not tested; NA = Not
active (>1000000 nM).
Pharmaceutical Compositions and Methods of Treatment
[0054] Compounds of formula I bind to alpha 7 and can be useful in
treating affective disorders and neurodegenerative disorders.
Therefore, another aspect of the invention is a composition
comprising a compound of formula I, or a pharmaceutically
acceptable salt thereof, and a pharmaceutically acceptable
carrier.
[0055] Another aspect of the invention is the use of a compound of
formula I in the manufacture of a medicament for the treatment of
affective disorders or neurodegenerative disorders.
[0056] Another aspect of the invention is the use of a compound of
formula I in the manufacture of a medicament for the treatment of
schizophrenia or Alzheimer's Disease.
[0057] Another aspect of the invention is a method of treating
affective disorders or neurodegenerative disorders comprising
administering to a patient a therapeutically effective amount of a
compound of formula I.
[0058] Another aspect of the invention is a method of treating
schizophrenia or Alzheimer's Disease comprising administering to a
patient a therapeutically effective amount of a compound of formula
I.
[0059] Another aspect of the invention is a method of treating
schizophrenia comprising administering to a patient a
therapeutically effective amount of a compound of formula I.
[0060] Another aspect of the invention is a method of treating
Alzheimer's Disease comprising administering to a patient a
therapeutically effective amount of a compound of formula I.
[0061] Another aspect of the invention is a method of treating
cognitive disorders comprising administering to a patient a
therapeutically effective amount of a compound of formula I.
[0062] Another aspect of the invention is a method of treating
rheumatoid arthritis comprising administering to a patient a
therapeutically effective amount of a compound of formula I.
[0063] Another aspect of the invention is a method of treating
osteoarthritis comprising administering to a patient a
therapeutically effective amount of a compound of formula I.
[0064] Another aspect of the invention is a method of treating
ulcerative colitis comprising administering to a patient a
therapeutically effective amount of a compound of formula I.
[0065] Another aspect of the invention is a method of treating
Crohn's Disease comprising administering to a patient a
therapeutically effective amount of a compound of formula I.
[0066] Another aspect of the invention is a method of treating
diabetes comprising comprising administering to a patient a
therapeutically effective amount of a compound of formula I.
[0067] "Patient" means a person suitable for therapy as understood
by practitioners in the field of affective disorders and
neurodegenerative disorders.
[0068] "Treatment," "therapy," and related terms are used as
understood by practitioners in the field of affective disorders and
neurodegenerative disorders.
[0069] The compounds of this invention are generally given as
pharmaceutical compositions comprised of a therapeutically
effective amount of a compound or its pharmaceutically acceptable
salt and a pharmaceutically acceptable carrier and may contain
conventional excipients. Pharmaceutically acceptable carriers are
those conventionally known carriers having acceptable safety
profiles. Compositions encompass all common solid and liquid forms
including for example capsules, tablets, losenges, and powders as
well as liquid suspensions, syrups, elixers, and solutions.
Compositions are made using common formulation techniques, and
conventional excipients (such as binding and wetting agents) and
vehicles (such as water and alcohols) are generally used for
compositions. See, for example, Remington's Pharmaceutical
Sciences, Mack Publishing Company, Easton, Pa., 17th edition,
1985.
[0070] Solid compositions are normally formulated in dosage units
and compositions providing from about 1 to 1000 mg of the active
ingredient per dose are preferred. Some examples of dosages are 1
mg, 10 mg, 100 mg, 250 mg, 500 mg, and 1000 mg. Generally, other
agents will be present in a unit range similar to agents of that
class used clinically. Typically, this is 0.25-1000 mg/unit.
[0071] Liquid compositions are usually in dosage unit ranges.
Generally, the liquid composition will be in a unit dosage range of
1-100 mg/mL. Some examples of dosages are 1 mg/mL, 10 mg/mL, 25
mg/mL, 50 mg/mL, and 100 mg/mL. Generally, other agents will be
present in a unit range similar to agents of that class used
clinically. Typically, this is 1-100 mg/mL.
[0072] The invention encompasses all conventional modes of
administration; oral and parenteral methods are preferred.
Generally, the dosing regimen will be similar to other agents used
clinically. Typically, the daily dose will be 1-100 mg/kg body
weight daily. Generally, more compound is required orally and less
parenterally. The specific dosing regime, however, will be
determined by a physician using sound medical judgement.
DESCRIPTION OF SPECIFIC EMBODIMENTS
[0073] .sup.1H-NMR spectra were run on a Bruker 500, 400, or 300
MHz instrument and chemical shifts were reported in ppm (.delta.)
with reference to tetramethylsilane (.delta.=0.0). All evaporations
were carried out under reduced pressure. Unless otherwise stated,
LC/MS analyses were carried out on a Shimadzu instrument using a
Phenomenex-Luna 4.6.times.50 mm S 10 reverse phase column employing
a flow rate of 4 mL/min using a 0.1% TFA in methanol/water gradient
[0-100% in 3 min, with 4 min run time] and a UV detector set at 220
nm or Gemini C18 4.6.times.50 mm 5 u reverse phase column employing
a flow rate of 5 mL/min using a 10 mM ammonium acetate
acetonitrile/water gradient [5-95% in 3 min, with 4 min run time]
and a UV detector set at 220 nm (negative-ion mass spectrometry).
Unless otherwise stated, purification could be done by preparative
C-18 column employing gradients of methanol-water containing 0.1%
of trifluoroacetic acid (TFA), and using a Shimadzu High
Performance Liquid Preparative Chromatographic System employing an
XTERRA 30.times.100 mm S5 column at 40 mL/min flow rate with a 12
min gradient.
Example 1
N-(benzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]--
2-amine
##STR00392##
[0074] Step A:
N-(Benzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
##STR00393##
[0076] To benzo[d]thiazol-2-amine (20 g, 133 mmol) in acetonitrile
(300 mL) was added 1,1'-thiocarbonyldiimidazole (30.8 g, 173 mmol).
The reaction was stirred at 50.degree. C. for 24 hours. The
reaction was cooled to room temperature and the precipitate was
filtered and washed with acetonitrile (2.times.50 mL). The yellow
powder was dried in a vacuum oven (40.degree. C.) for 2 hours. The
product,
N-(benzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide (28.9
g, 111 mmol), was taken directly to the next step without any
further purification.
Step B:
(3-((3-Benzo[d]thiazol-2-ylthioureido)methyl)-3-hydroxy-1-ammoniob-
icyclo[2.2.2]octan-1-yl)trihydroborate
##STR00394##
[0078] To
N-(benzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide (9.2 g,
35 mmol) in N,N-dimethylformamide (100 mL) was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (6.0 g, 35 mmol) which was synthesized according to Swain C. J.,
et. al., J. Med. Chem., 35:1019-1031 (1992). The reaction was
stirred at 65.degree. C. for 15 hours. The reaction was cooled and
concentrated to yield the crude product. The crude material was
purified via flash chromatography (50-100% ethyl acetate/hexanes)
yielding the first spot/fractions detected by TLC as the product.
The fractions were combined and concentrated to yield
(3-((3-benzo[d]thiazol-2-ylthioureido)methyl)-3-hydroxy-1-ammoniobicyclo[-
2.2.2]octan-1-yl)trihydroborate (10.6 g, 29.1 mmol, 83% yield) as
an off-white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 11.88 (s, 1H), 10.30 (s, 1H), 7.94 (d, J=7.63 Hz, 1H),
7.55-7.74 (m, 1H), 7.37-7.53 (m, J=7.32, 7.32 Hz, 1H), 7.16-7.37
(m, J=7.63, 7.63 Hz, 1H), 5.39 (s, 1H), 3.85 (d, 2H), 2.65-3.08 (m,
6H), 1.99-2.22 (m, 1H), 1.79-1.97 (m, 2H), 1.66-1.79 (m, 1H),
1.08-1.63 (m, 4H). MS (LC/MS) R.T.=3.40; [M+H].sup.+=363.1
Step C:
(2-(Benzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bic-
yclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00395##
[0080] To
(3-((3-benzo[d]thiazol-2-ylthioureido)methyl)-3-hydroxy-1-ammoni-
obicyclo[2.2.2]octan-1-yl)trihydroborate (10.6 g, 29.1 mmol) in
N,N-dimethylformamide (100 mL) was added
N,N'-diisopropylcarbodiimide (11.4 mL, 72.8 mmol). The reaction was
stirred at 70.degree. C. for 4 hours. The reaction was concentrated
to yield a crude residue. A small amount of ethyl acetate (20 mL)
was added and the suspension was sonicated. The solids were
filtered and washed with small portions of ethyl acetate
(2.times.10 mL). The solids were dried in a vacuum oven (80.degree.
C.) to yield
(2-(benzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bicyclo[2.-
2.2]octane]-1'-yl)trihydroborate (6.83 g, 20.8 mmol, 72% yield) as
a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
9.09 (br. s., 1H) 7.81 (d, J=7.63 Hz, 1H) 7.63 (d, J=7.93 Hz, 1H)
7.30-7.40 (m, 1H) 7.15-7.24 (m, 1H) 3.88 (d, J=10.38 Hz, 1H) 3.77
(d, J=10.38 Hz, 1H) 3.25-3.37 (m, 1H) 3.17 (dd, J=14.95, 1.83 Hz,
1H) 2.99-3.10 (m, 1H) 2.79-2.98 (m, 3H) 2.27 (br. s., 1H) 1.98-2.11
(m, 1H) 1.71-1.88 (m, 3H) 1.45 (br. s., 3H). MS (LC/MS) R.T.=2.44;
[M+H-BH.sub.3].sup.+=315.1.
Step D:
N-(Benzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00396##
[0082] To
(2-(benzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-b-
icyclo[2.2.2]octane]-1'-yl)trihydroborate (6.6 g, 20.1 mmol) in
acetone (9 mL) was added 3M HCl (50.3 mL, 151 mmol). The reaction
was stirred at room temperature for 4 hours. The reaction was
complete by TLC (lower spot). Ethyl acetate was added and the
aqueous layer was then separated. The aqueous layer was neutralized
with 1N sodium hydroxide. The product was extracted with ethyl
acetate (2.times.150 mL). The organics were combined, dried with
magnesium sulfate, filtered, and concentrated in vacuo to afford a
white powder. A small amount of ethyl acetate (20 mL) was added to
the powder. The solids were sonicated and filtered to yield racemic
N-(benzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (5.13 g, 16.3 mmol, 81% yield) as a white powder.
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.02 (br. s., 1H)
7.79 (d, J=7.02 Hz, 1H) 7.62 (d, J=7.63 Hz, 1H) 7.29-7.38 (m, 1H)
7.15-7.22 (m, 1H) 3.90 (d, J=10.07 Hz, 1H) 3.65 (d, J=10.07 Hz, 1H)
2.98-3.10 (m, 2H) 2.73-2.88 (m, 2H) 2.67 (t, J=7.78 Hz, 2H) 2.07
(br. s., 1H) 1.93 (br. s., 1H) 1.42-1.67 (m, 3H). MS (LC/MS)
R.T.=1.15; [M+H].sup.+=315.3.
[0083] The enantiomers were separated using a Chiralpak AD-H
(3.times.25 cm, 5 uM) column with a mobile phase consisting of
CO.sub.2/(methanol/ACN/DEA=70/30/0.1 (v/v/v))=77/23. The wavelength
was set at 300 nM. The separated peaks were concentrated in vacuo
to yield white powders. The first peak off the column was
(S)--N-(benzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]o-
ctan]-2-amine (1.45 g, 4.61 mmol, 29.4% yield). (1a; S-isomer):
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.03 (br. s., 1H)
7.78 (d, J=7.05 Hz, 1H) 7.61 (d, J=7.55 Hz, 1H) 7.27-7.37 (m, 1H)
7.11-7.23 (m, 1H) 3.89 (d, J=10.07 Hz, 1H) 3.64 (d, J=10.07 Hz, 1H)
2.96-3.09 (m, 2H) 2.71-2.88 (m, 2H) 2.66 (t, J=7.81 Hz, 2H)
2.02-2.11 (m, 1H) 1.85-1.97 (m, 1H) 1.41-1.65 (m, 3H). MS (LC/MS)
R.T.=1.15; [M+H].sup.+=315.3. Optical rotation (1.23 mg/mL,
DMSO)=+5.20.degree.. The second peak was
(R)--N-(benzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]o-
ctan]-2-amine (1.21 g, 3.85 mmol, 24.5% yield). (1b; R-isomer):
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.02 (br. s., 1H)
7.79 (d, J=7.32 Hz, 1H) 7.62 (d, J=7.63 Hz, 1H) 7.33 (t, J=7.63 Hz,
1H) 7.18 (t, J=7.48 Hz, 1H) 3.90 (d, J=10.07 Hz, 1H) 3.65 (d,
J=10.07 Hz, 1H) 2.98-3.10 (m, 2H) 2.73-2.87 (m, 2H) 2.67 (t, J=7.63
Hz, 2H) 2.08 (br. s., 1H) 1.93 (br. s., 1H) 1.42-1.67 (m, 3H). MS
(LC/MS) R.T.=1.15; [M+H].sup.+=315.3; Optical rotation (3.9 mg/mL,
DMSO)=-3.92.degree..
Example 2
N-(5-Methoxythiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00397##
[0084] Step A: 5-Methoxythiazolo[5,4-b]pyridin-2-amine
##STR00398##
[0086] In a 500 ml, 3-neck flask equipped with a mechanical
stirrer, dropping funnel and thermometer, acetic acid (100 mL) was
added and cooled in an ice bath. Potassium thiocyanate (40 g, 412
mmol) and 6-methoxypyridin-3-amine (6.2 g, 49.9 mmol) were added to
the reaction mixture. The reaction was cooled in an ice-salt bath
until the reaction temperature reached <0.degree. C. A solution
of bromine (8 mL, 156 mmol) in acetic acid (30.0 mL) was added
dropwise over 2 hours at a rate that maintained the reaction
temperature <0.degree. C. Mechanical stirring was required.
After the addition was complete, the mixture was left to stir and
allowed to slowly warm to room temperature overnight. Water (30 mL)
was then added and the mixture was heated to 85.degree. C. in an
oil bath. This mixture was then filtered while still hot. The
orange filter cake was returned to the reaction flask, and an
additional 50 ml acetic acid was added. The mixture was heated
again to 85.degree. C., and then filtered while still hot once
more. The combined filtrates were cooled in an ice bath and
neutralized to pH 8 with conc. ammonium hydroxide. A purple
precipitate formed which was then collected by filtration to afford
5 g of crude material. This crude material was recrystallized from
methanol (40 mL) to yield 5-methoxythiazolo[5,4-b]pyridin-2-amine
(3 g, 16.55 mmol, 33.1% yield) as purple crystals. .sup.1H NMR (300
MHz, DMSO-d.sub.6) .delta. ppm 7.60 (1H, d, J=8.42 Hz), 7.41 (2H,
s), 6.67 (1H, d, J=8.78 Hz), 3.81 (3H, s).
Step B:
(3-Hydroxy-3-((3-(5-methoxythiazolo[5,4-b]pyridin-2-yl)thioureido)-
methyl)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00399##
[0088] 5-Methoxythiazolo[5,4-b]pyridin-2-amine (2.4 g, 13.24 mmol)
was divided among 5.times.20 mL screw-cap vials. To each vial was
added acetonitrile (10 mL) and thiocarbonyl diimidazole (600 mg).
All vials were heated at 60.degree. C. overnight. The reaction
vials were combined and concentrated to yield crude
N-(5-methoxythiazolo[5,4-b]pyridin-2-yl)-1H-imidazole-1-carbothioamide
product.
[0089] This crude product was suspended in N,N-dimethylformamide
(50 ml) and
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydro-
borate (2.7 g, 15.88 mmol) was then added. The reaction was heated
at 70.degree. C. for 4 hours. LC/MS showed essentially complete
conversion. The reaction was cooled to room temperature and then
poured into water. The product was extracted first with toluene,
and then with chloroform. The organics were combined, washed with
water and brine, dried over sodium sulfate, filtered and
concentrated in vacuo to afford crude material. This crude material
was purified via flash chromatography (20-100% ethyl
acetate-hexane). The product fractions were collected and
concentrated in vacuo to afford
(3-hydroxy-3-((3-(5-methoxythiazolo[5,4-b]pyridin-2-yl)thioureido)methyl)-
-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (1.36 g, 3.46
mmol, 26.1% yield). .sup.1HNMR showed a 1:0.55 molar ratio of
(3-hydroxy-3-((3-(5-methoxythiazolo[5,4-b]pyridin-2-yl)thioureido)methyl)-
-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (1.36 g, 3.46
mmol, 26.1% yield) to 5-methoxythiazolo[5,4-b]pyridin-2-amine (0.34
g, 1.876 mmol, 14.17% yield). The mixture was taken directly to the
next step without any further purification. MS (LC/MS) R.T.=3.23;
[M+H].sup.+=392.1.
Step C:
(2-(5-Methoxy-3a,7a-dihydrothiazolo[5,4-b]pyridin-2-ylamino)-4H-1'-
-ammoniospiro[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00400##
[0091] To
(3-hydroxy-3-((3-(5-methoxythiazolo[5,4-b]pyridin-2-yl)thioureid-
o)methyl)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (1.7 g,
3.46 mmol) in N,N-dimethylformamide (10 mL) was added
N,N'-diisopropylcarbodiimide (1.89 mL, 12.10 mmol). The reaction
was stirred at 70.degree. C. for 2 hours. The mixture was cooled
and then poured into water. The product was extracted with toluene
and chloroform. The organic layers were combined, washed with
brine, dried over sodium sulfate, filtered and concentrated in
vacuo to afford crude material. This solid material was triturated
with ether. The solids were then filtered and dried to yield
(2-(5-methoxy-3a,7a-dihydrothiazolo[5,4-b]pyridin-2-ylamino)-4H-1'-ammoni-
ospiro[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (700
mg, 1.94 mmol, 56.0% yield). .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. ppm 8.98 (1H, s), 7.86 (1H, d, J=8.78 Hz), 6.81 (1H, d,
J=8.42 Hz), 3.87 (3H, s), 3.83 (1H, s), 3.68-3.78 (1 H, m), 3.31
(1H, s), 3.15-3.29 (1H, m), 2.78-3.14 (4H, m), 2.26 (1H, br. s.),
2.04 (1H, br. s.), 1.63-1.89 (3H, m).
Step D:
N-(5-Methoxythiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine
##STR00401##
[0093] To
(2-(5-methoxythiazolo[5,4-b]pyridin-2-ylamino)-4H-1'-ammoniospir-
o[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (780 mg,
2.17 mmol) in acetone (10 mL) was added 3M HCl (10 mL, 329 mmol).
The reaction was stirred at room temperature for 2 hours.
Chloroform and water were added and the aqueous layer was then
separated. The aqueous layer was neutralized with sodium
bicarbonate. The product was extracted with chloroform (2.times.).
The organics were combined, dried over sodium sulfate, filtered and
concentrated in vacuo to afford racemic
N-(5-methoxythiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine (588 mg, 1.70 mmol, 78% yield) as a white
powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.90 (1H,
br. s.), 7.82-7.86 (1H, m), 6.80 (1H, d, J=8.85 Hz), 3.84-3.89 (4H,
m), 3.61 (1H, d, J=10.07 Hz), 3.03 (2H, d, J=5.19 Hz), 2.73-2.86
(2H, m), 2.66 (2H, t, J=7.78 Hz), 2.07 (1H, br. s.), 1.92 (1H, br.
s.), 1.45-1.65 (3H, m). MS (LC/MS) R.T.=1.29;
[M+H].sup.+=346.1.
[0094] The enantiomers were separated using a Chiralpak AD-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
35% methanol (0.1% DEA) in CO.sub.2. The wavelength was set at 300
nM. The separated peaks were concentrated in vacuo to yield white
powders. The first peak off the column was
(S)--N-(5-methoxythiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octan]-2-amine (212 mg, 0.61 mmol, 36.1% yield).
(2a; S-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
8.89 (1H, br. s.), 7.84 (1H, d, J=8.85 Hz), 6.77-6.82 (1H, m),
3.84-3.89 (4H, m), 3.61 (1H, d, J=10.07 Hz), 3.03 (2H, d, J=5.19
Hz), 2.74-2.86 (2H, m), 2.66 (2H, t, J=7.63 Hz), 2.06 (1H, br. s.),
1.92 (1H, br. s.), 1.44-1.65 (3H, m). MS (LC/MS) R.T.=1.47;
[M+H].sup.+=346.2. Optical rotation (3.57 mg/ml,
DMSO)=-2.58.degree.. The second peak was
(R)--N-(5-methoxythiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octan]-2-amine (242 mg, 0.70 mmol, 41.2% yield).
(2b; R-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
8.89 (1H, br. s.), 7.84 (1H, d, J=8.85 Hz), 6.79 (1H, d, J=8.55
Hz), 3.82-3.90 (4H, m), 3.61 (1H, d, J=10.07 Hz), 3.03 (2H, d,
J=5.49 Hz), 2.74-2.86 (2H, m), 2.66 (2H, t, J=7.78 Hz), 2.06 (1H,
br. s.), 1.92 (1 H, br. s.), 1.42-1.67 (3H, m). MS (LC/MS)
R.T.=1.30; [M+H].sup.+=346.2. Optical rotation (3.29 mg/ml,
DMSO)=+2.43.degree..
Example 3
(2-(5H-1'-Azaspiro[oxazole-4,3'-bicyclo[2.2.2]octane]-2-ylamine)benzo[d]th-
iazol-6-yl)pyrrolidin-1-yl)methanone
##STR00402##
[0095] Step A: tert-Butyl
6-(pyrrolidine-1-carbonyl)benzo[d]thiazol-2-ylcarbamate
##STR00403##
[0097] In a 250 ml flask was added
2-(tert-butoxycarbonyl-amino)benzo[d]thiazole-6-carboxylic acid
(1.0 g, 3.4 mmol) and pyrrolidine (0.559 mL, 6.8 mmol) in
tetrahydrofuran (50 mL). To this solution was added EDC (1.3 g, 6.8
mmol), 1-hydroxybenzotriaxole (1.041 g, 6.8 mmol) and Hunig's Base
(2.37 mL, 13.59 mmol). The reaction was stirred at 25.degree. C.
for 1 hour. The reaction was then poured into water and
dichloromethane. The water was extracted 3 times with
dichloromethane and the organic layers were combined and
concentrated. The residue was taken up in a small amount of
dichloromethane and precipitated out with diethyl ether/hexanes.
The flask was put in the freezer for 1 hour and filtered. The white
precipitate was collected to yield tert-butyl
6-(pyrrolidine-1-carbonyl)benzo[d]thiazol-2-ylcarbamate (1.09 g,
3.14 mmol, 92% yield). .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 11.95 (s, 1H), 8.14 (s, 1H), 7.61-7.77 (m, J=8.39, 1.98 Hz,
1H), 7.46-7.63 (m, 1H), 3.39-3.63 (m, 4H), 1.74-2.00 (m, 4H),
1.45-1.62 (m, 9H). MS (LC/MS) R.T.=3.40; [M+H].sup.+=363.1.
Step B: (2-Aminobenzo[d]thiazol-6-yl)(pyrrolidin-1-yl)methone
##STR00404##
[0099] tert-Butyl
6-(pyrrolidine-1-carbonyl)benzo[d]thiazol-2-ylcarbamate (1.09 g,
3.14 mmol) was dissolved in dichloromethane (10 mL) and TFA (3 mL,
38.9 mmol) and the reaction was stirred at 25.degree. C. overnight.
The reaction was poured into a separatory funnel and carefully
neutralized with sodium bicarbonate. The liquid was extracted 3
times with chloroform/methanol (4:1). The organic layers were
concentrated to a white residue and triturated in diethyl ether.
The precipitate was collected to yield
(2-aminobenzo[d]thiazol-6-yl)(pyrrolidin-1-yl)methone (0.497 g, 2.0
mmol, 64%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.86
(d, J=1.51 Hz, 1H), 7.65 (s, 2H), 7.35-7.46 (m, 1H), 7.26-7.35 (m,
1H), 3.41-3.57 (m, 4H), 1.64-1.97 (m, 4H). MS (LC/MS) R.T.=1.39;
[M+H].sup.+=248.1.
Step C:
(2-(5H-1'-Azaspiro[oxazole-4,3'-bicyclo[2.2.2]octane]-2-ylamine)be-
nzo[d]thiazol-6-yl)pyrrolidin-1-yl)methanone
##STR00405##
[0101]
(2-(5H-1'-Azaspiro[oxazole-4,3'-bicyclo[2.2.2]octane]-2-ylamine)ben-
zo-[d]thiazol-6-yl)pyrrolidin-1-yl)methanone was prepared by
following the general procedures of Example 1, Steps A-D and using
(2-aminobenzo[d]thiazol-6-yl)(pyrrolidin-1-yl)methone (Example 3,
Step B) as the starting material. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.05 (br. s., 1H) 7.99 (d, J=1.76 Hz, 1H)
7.60 (d, J=8.31 Hz, 1H) 7.48 (dd, J=8.31, 1.76 Hz, 1H) 3.90 (d,
J=10.07 Hz, 1H) 3.65 (d, J=10.07 Hz, 1H) 3.42-3.52 (m, 4H) 3.04 (s,
2H) 2.75-2.87 (m, 2H) 2.67 (t, J=7.81 Hz, 2H) 2.08 (br. s., 1H)
1.76-1.98 (m, 5H) 1.41-1.65 (m, 3H). MS (LC/MS) R.T.=1.33;
[M+H].sup.+=412.2.
Example 4
N-(5-Phenylthiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]--
2-amine
##STR00406##
[0102] Step A:
(3-Hydroxy-3-((3-(5-phenylthiazol-2-yl)thioureido)methyl)-1-ammoniobicycl-
o[2.2.2]octan-1-yl)trihydroborate
##STR00407##
[0104] To 5-phenylthiazol-2-amine (0.52 g, 2.9 mmol) in
acetonitrile (6 mL) was added 1,1'-thiocarbonyldiimidazole (0.68 g,
3.8 mmol). The reaction mixture was stirred at 65.degree. C. for 2
hours. The precipitate was filtered and washed with acetonitrile
(2.times.20 mL) to yield intermediate
N-(5-phenylthiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide. The
intermediate was taken up in N,N-dimethylformamide (30 mL) and
treated with
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydr-
oborate (0.5 g, 2.9 mmol). The reaction mixture was stirred for 5
hours at 65.degree. C. The reaction was concentrated in vacuo and
purified via silica gel chromatography (30-100% ethyl
acetate/hexane). The product fractions were combined and
concentrated in vacuo to yield
(3-hydroxy-3-((3-(5-phenylthiazol-2-yl)thioureido)methyl)-1-ammoniobicycl-
o[2.2.2]octan-1-yl)trihydroborate (0.85 g, 2.19 mmol, 74.4% yield)
as a white powder. LC/MS confirmed product with loss of BH.sub.3 in
the LC/MS conditions: retention time 3.26
(M+1-BH.sub.3=375.33).
Step B:
(2-(5-Phenylthiazol-2-ylamino)-4H-1'-ammoniospiro-[oxazole-5,3'-bi-
cyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00408##
[0106] To
(3-hydroxy-3-((3-(5-phenylthiazol-2-yl)thioureido)methyl)-1-ammo-
niobicyclo[2.2.2]octan-1-yl)trihydroborate (0.8 g, 2.1 mmol) in
N,N-dimethylformamide (20 mL) was added
N,N'-diisopropylcarbodiimide (1.12 mL, 7.2 mmol). The reaction
mixture was stirred at 70.degree. C. for 4 hours. The reaction was
concentrated in vacuo and purified via silica gel chromatography
(40-100% ethyl acetate/hexane). The combined product fractions were
concentrated in vacuo to yield
(2-(5-phenylthiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bicyclo[2.-
2.2]octane]-1'-yl)trihydroborate (0.51 g, 1.44 mmol, 70% yield) as
a white powder. LCMS-mass corresponds to BH.sub.3 lost in the LC/MS
conditions: retention time 2.46 (M+1-BH.sub.3=341.36).
Step C:
N-(5-Phenylthiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00409##
[0108] To
(2-(5-phenylthiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-b-
icyclo[2.2.2]octane]-1'-yl)trihydroborate (0.56 g, 1.58 mmol) in
acetone (9 mL) was added 3 M HCl (3.95 mL, 11.86 mmol). The
reaction mixture was stirred at room temperature for 4 hours and
then neutralized with 1N sodium hydroxide. The product was
extracted with ethyl acetate (2.times.20 mL), followed by
chloroform (2.times.20 mL). The organics were combined, dried with
magnesium sulfate, filtered, and concentrated in vacuo to afford a
white powder. The crude product was purified by reverse phase HPLC
(Phenomenex Luna 30.times.100 mm; 220 wavelength; gradient time 10
min; flow rate 40 ml/min; solvent A; 10% methanol-90% water-0.1%
TFA, solvent B; 90% methanol-10% water-0.1% TFA). The fractions
were combined, neutralized with 1N sodium hydroxide and extracted
with ethyl acetate (2.times.30 mL) and chloroform (2.times.30 mL).
The organics were combined, dried with magnesium sulfate, filtered
and concentrated in vacuo to yield
N-(5-phenylthiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-
-2-amine (0.4 g, 1.175 mmol, 74.3% yield) as a white powder.
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.63 (1H, br. s.),
7.71 (1H, s), 7.52 (2H, d, J=7.32 Hz), 7.37 (2H, t, J=7.78 Hz),
7.25 (1H, t, J=7.32 Hz), 3.82 (1H, d, J=10.07 Hz), 3.57 (1H, d,
J=9.77 Hz), 3.02 (2H, d, J=4.27 Hz), 2.79 (2H, t, J=7.63 Hz), 2.66
(2H, t, J=7.63 Hz), 2.04 (1H, br. s.), 1.92-1.97 (1H, m), 1.44-1.65
(3H, m). MS (LC/MS) R.T.=1.52; [M+H].sup.+=341.3.
[0109] The enantiomers were separated using a Chiralpak AS-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
30% methanol (0.1% DEA) in CO.sub.2 and UV monitored at 300 nm. The
separated peaks were concentrated in vacuo to yield white powders.
The first peak off the column yielded 0.4 g, 1.15 mmol, 32.7% (4a,
S-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.69 (s,
1H), 7.69-7.87 (m, 1H), 7.52-7.66 (m, J=10.99 Hz, 2H), 7.37-7.51
(m, 2H), 7.21-7.38 (m, 2H), 3.52-4.00 (m, 2H), 2.98-3.23 (m, 2H),
2.78-2.94 (m, 2H), 2.64-2.78 (m, 2H), 1.88-2.19 (m, J=60.43 Hz,
2H), 1.40-1.77 (m, 3H). MS (LC/MS) R.T.=1.77; [M+H].sup.+=341.1.
The second peak yielded 0.4 g, 1.15 mmol, 32.7%. (4b, R-isomer): L
M.P. 187-9.degree. C. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 8.64 (s, 1H), 7.62-7.84 (m, 1H), 7.44-7.61 (m, 2H), 7.33-7.46
(m, 2H), 7.18-7.31 (m, 1H), 3.50-3.99 (m, 2H), 2.94-3.14 (m, 2H),
2.74-2.91 (m, 2H), 2.61-2.72 (m, 2H), 2.05 (s, 1H), 1.82-2.00 (m,
1H), 1.34-1.72 (m, 3H). MS (LC/MS) R.T.=1.78;
[M+H].sup.+=341.1.
Example 5
N-(6-Methoxybenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
Step A:
N-(6-Methoxybenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioami-
de
##STR00410##
[0111] To 6-methoxybenzo[d]thiazol-2-amine (0.53 g, 2.94 mmol) in
acetonitrile (20 mL) was added 1,1'-thiocarbonyldiimidazole (0.681
g, 3.82 mmol). The reaction mixture was stirred at 65.degree. C.
for 24 hours. The precipitate was filtered and washed with
acetonitrile (2.times.20 mL) to yield the product. The product was
taken directly to the next step without any further purification or
characterization.
Step B:
(3-Hydroxy-3-((3-(6-methoxybenzo[d]thiazol-2-yl)-thioureido)methyl-
)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00411##
[0113] To
N-(6-methoxybenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioa-
mide (0.82 g, 2.82 mmol) in N,N-dimethylformamide (20 mL) was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (0.48 g, 2.82 mmol). The reaction mixture was stirred at
65.degree. C. for 6 hours. The reaction was concentrated in vacuo
and then purified by silica gel chromatography (30%-100% ethyl
acetate/hexanes). The pure fractions were combined and concentrated
to yield
(3-hydroxy-3-((3-(6-methoxybenzo[d]thiazol-2-yl)thioureido)methyl)-1-ammo-
niobicyclo[2.2.2]octan-1-yl)trihydroborate (0.7 g, 1.78 mmol, 63.2%
yield) as a white powder. LC/MS confirmed product as loss of
BH.sub.3 in the LC/MS conditions: retention time 3.11
(M+1-BH.sub.3=379.4).
Step C:
(2-(6-Methoxybenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-
-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00412##
[0115] To
(3-hydroxy-3-((3-(6-methoxybenzo[d]thiazol-2-yl)thioureido)methy-
l)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (0.68 g, 1.73
mmol) in N,N-dimethylformamide (20 mL) was added
N,N'-diisopropylcarbodiimide (0.95 mL, 6.1 mmol). The reaction
mixture was stirred at 70.degree. C. for 4 hours. The reaction was
concentrated and purified via silica gel chromatography (40-100%
ethyl acetate/hexane). The product fractions were combined and
concentrated in vacuo to yield
(2-(6-methoxybenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-b-
icyclo[2.2.2]octane]-1'-yl)trihydroborate (0.35 g, 0.98 mmol, 56.4%
yield) as a white powder. LC/MS MH.sup.+-BH.sub.3=345.2.
Step D:
N-(6-Methoxybenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00413##
[0117] To
(2-(6-methoxybenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazo-
le-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.33 g, 0.921
mmol) in acetone (9 mL) was added 3 M HCl (2.30 mL, 6.91 mmol). The
reaction was stirred at room temperature for 4 hours and then
neutralized with 1N sodium hydroxide. The product was extracted
with ethyl acetate (2.times.20 mL), followed by chloroform
(2.times.20 mL). The organics were combined, dried with magnesium
sulfate, filtered, and concentrated in vacuo to afford a white
powder. The crude product was purified by reverse phase HPLC
(Phenomenex Luna 30.times.100 mm; 220 wavelength; gradient time 10
min; flow rate 40 ml/min; solvent A; 10% methanol-90% water-0.1%
TFA, solvent B; 90% methanol-10% water-0.1% TFA). The fractions
were combined, neutralized with 1N sodium hydroxide and extracted
with ethyl acetate (2.times.30 mL). The organics were combined,
dried with magnesium sulfate, filtered and concentrated in vacuo to
yield
N-(6-methoxybenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2-
.2]octan]-2-amine (0.25 g, 0.73 mmol, 79% yield) as a white powder.
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.88 (1H, d, J=1.22
Hz), 7.48-7.52 (1H, m), 7.40 (1H, d, J=2.75 Hz), 6.92 (1H, dd,
J=8.70, 2.59 Hz), 3.87 (1H, d, J=9.77 Hz), 3.77 (3H, s), 3.61 (1H,
d, J=9.77 Hz), 3.03 (2H, s), 2.75-2.86 (2H, m), 2.67 (2H, t, J=7.78
Hz), 2.06 (1H, br. s.), 1.91 (1H, br. s.), 1.41-1.65 (3H, m). MS
(LC/MS) R.T.=1.44; [M+H].sup.+=345.3.
[0118] The enantiomers were separated using a Chiralpak AD-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
23% methanol (0.1% DEA) in CO.sub.2 and UV monitored at 220 nm. The
separated peaks were concentrated in vacuo to yield white powders.
The first peak off the column yielded 205.5 mg, 0.60 mmol, 34.1%.
(5a, S-isomer): .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
8.88 (1H, br. s.), 7.50 (1H, d, J=8.81 Hz), 7.40 (1H, d, J=2.52
Hz), 6.92 (1H, dd, J=8.81, 2.77 Hz), 3.87 (1H, d, J=9.82 Hz), 3.77
(3H, s), 3.58-3.65 (1H, m), 3.02 (2H, s), 2.74-2.86 (2H, m), 2.66
(2H, t, J=7.68 Hz), 2.03-2.08 (1H, m), 1.91 (1H, br. s.), 1.39-1.65
(3H, m). MS (LC/MS) R.T.=1.40; [M+H].sup.+=345.2. The second peak
yielded 206.9 mg, 0.6 mmol, 34%. (5b, R-isomer): .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 8.88 (1H, br. s.), 7.50 (1H, d,
J=8.56 Hz), 7.40 (1H, d, J=2.52 Hz), 6.93 (1H, dd, J=8.81, 2.52
Hz), 3.87 (1H, d, J=10.07 Hz), 3.77 (3H, s), 3.62 (1H, d, J=10.07
Hz), 3.02 (2H, s), 2.75-2.86 (2H, m), 2.67 (2H, t, J=7.68 Hz),
2.04-2.09 (1H, m), 1.92 (1H, br. s.), 1.42-1.66 (3H, m). MS (LC/MS)
R.T.=1.70; [M+H].sup.+=345.1.
Example 6
N-(4-Methylbenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00414##
[0119] Step A:
N-(4-Methylbenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
##STR00415##
[0121] To 4-methylbenzo[d]thiazol-2-amine (1.1 g, 6.7 mmol) in
acetonitrile (30 mL) was added 1,1'-thiocarbonyldiimidazole (1.552
g, 8.71 mmol). The reaction mixture was stirred at 50.degree. C.
for 18 hours. The reaction was cooled to room temperature and the
precipitate was filtered and washed with acetonitrile (2.times.50
mL). The yellow powder was dried in a vacuum oven (50.degree. C.)
for 1 hour to yield
N-(4-methylbenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
(900 mg, 3.28 mmol, 49% yield) and then used in the next step
without any further purification or characterization.
Step B:
(3-Hydroxy-3-((3-(4-methylbenzo[d]thiazol-2-yl)thioureido)methyl)--
1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00416##
[0123] To
N-(4-methylbenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioam-
ide (0.71 g, 2.59 mmol) in N,N-dimethylformamide (20 mL) was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (0.44 g, 2.59 mmol). The reaction mixture was stirred at
70.degree. C. for 4 hours. The reaction was concentrated and
purified via silica gel chromatography (40-100% ethyl
acetate/hexanes). The product fractions were concentrated in vacuo
to yield
(3-hydroxy-3-((3-(4-methylbenzo[d]thiazol-2-yl)thioureido)methyl)-1-ammon-
iobicyclo[2.2.2]octan-1-yl)trihydroborate (0.65 g, 1.73 mmol, 66.7%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 11.93 (1H, s), 7.74 (1H, d, J=7.63 Hz), 7.22-7.26 (1 H,
m), 7.19 (1H, t, J=7.63 Hz), 3.75-3.93 (2H, m), 2.72-2.95 (6H, m),
2.56 (3H, s), 2.03-2.14 (1H, m), 1.95 (1H, br. s.), 1.78-1.87 (1H,
m), 1.73 (1H, ddd, J=13.81, 9.23, 5.04 Hz), 1.56 (1H, td, J=9.99,
7.78 Hz), 1.38 (2H, br. s.). (LC/MS) R.T.=3.70;
[M+H].sup.+=375.2.
Step C:
(2-(4-Methylbenzo[d]thiazol-2-ylamino)-4H-1'-ammonio-spiro[oxazole-
-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00417##
[0125] To
(3-hydroxy-3-((3-(4-methylbenzo[d]thiazol-2-yl)thioureido)methyl-
)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (0.62 g, 1.65
mmol) in N,N-dimethylformamide (20 mL) was added
N,N'-diisopropylcarbodiimide (0.33 mL, 2.14 mmol). The reaction
mixture was stirred at 70.degree. C. for 4 hours. The reaction was
concentrated and purified via silica gel chromatography (40-100%
ethyl acetate/hexanes). The product fractions were concentrated in
vacuo to yield
(2-(4-methylbenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bi-
cyclo[2.2.2]octane]-1'-yl)trihydroborate (0.4 g, 1.17 mmol, 70.9%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 8.90 (s, 1H), 7.61 (d, J=7.63 Hz, 1H), 7.13-7.17 (m,
1H), 7.09 (t, J=7.48 Hz, 1H), 3.90 (d, J=10.38 Hz, 1H), 3.77 (d,
J=10.38 Hz, 1H), 3.32 (s, 3H), 3.30 (d, J=1.53 Hz, 1H), 3.13-3.20
(m, 2 H), 3.00-3.09 (m, 1H), 2.85-2.94 (m, 4H), 2.57 (s, 4H), 2.28
(s, 1H), 2.06 (s, 1 H), 1.75-1.83 (m, 4H), 1.45 (s, 1H). (LC/MS)
R.T.=2.73; [M+H].sup.+=343.2.
Step D:
N-(4-Methylbenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00418##
[0127] To
(2-(4-methylbenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazol-
e-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.38 g, 1.11
mmol) in acetone (9 mL) was added 3 M HCl (2.78 mL, 8.33 mmol). The
reaction mixture was stirred at room temperature for 4 hours. Ethyl
acetate was added and the aqueous layer was collected and
neutralized with 1N sodium hydroxide. The product was extracted
with ethyl acetate (2.times.40 mL). The organics were combined,
dried with magnesium sulfate, filtered and concentrated in vacuo to
afford
N-(4-methylbenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (0.195 g, 0.594 mmol, 53.5% yield) as a white
powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.84 (s, 1
H), 7.60 (d, J=7.32 Hz, 1H), 7.13-7.17 (m, 1H), 7.08 (t, J=7.63 Hz,
1H), 3.92 (d, J=10.07 Hz, 1H), 3.66 (d, J=9.77 Hz, 1H), 3.04 (s,
2H), 2.76-2.85 (m, 2H), 2.68 (t, J=7.48 Hz, 2H), 2.56 (s, 3H), 2.09
(s, 1H), 1.93 (s, 1H), 1.61 (d, J=3.05 Hz, 1 H), 1.60 (s, 1H), 1.50
(dd, J=7.17, 2.59 Hz, 1H). (LC/MS) R.T.=1.76;
[M+H].sup.+=329.2.
[0128] The enantiomers were separated using a Chiralcel OJ-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
30% methanol (0.1% DEA) in CO.sub.2 and UV monitored at 300 nm. The
separated peaks were concentrated in vacuo to yield white powders.
The first peak off the column yielded 0.07 g, 0.21 mmol, 38.9%.
(6a, S-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
8.83 (br. s., 1H) 7.59 (d, J=7.63 Hz, 1H) 7.11-7.18 (m, 1H) 7.08
(t, J=7.63 Hz, 1H) 3.92 (d, J=10.38 Hz, 1H) 3.66 (d, J=9.77 Hz, 1H)
3.04 (s, 2H) 2.73-2.89 (m, 2H) 2.61-2.72 (m, 2H) 2.56 (s, 3H) 2.09
(br. s., 1H) 1.93 (br. s., 1H) 1.43-1.71 (m, 3H). MS (LC/MS)
R.T.=1.75; [M+H].sup.+=329.1. The second peak yielded 0.07 g, 0.21
mmol, 38.1%. (6b, R-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 8.84 (br. s., 1H) 7.59 (d, J=7.63 Hz, 1H) 7.15 (d,
J=7.20 Hz, 1H) 7.08 (t, J=7.48 Hz, 1H) 3.92 (d, J=9.77 Hz, 1H) 3.66
(d, J=10.07 Hz, 1H) 3.00-3.09 (m, 2H) 2.73-2.87 (m, 2H) 2.62-2.72
(m, 2H) 2.56 (s, 3H) 2.05-2.12 (m, 1H) 1.93 (br. s., 1H) 1.56-1.67
(m, 2H) 1.45-1.55 (m, 1H). MS (LC/MS) R.T.=1.75;
[M+H].sup.+=329.1.
Example 7
N-(4-Chlorobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00419##
[0129] Step A:
N-(4-Chlorobenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
##STR00420##
[0131] To 4-chlorobenzo[d]thiazol-2-amine (1.12 g, 6.07 mmol) in
acetonitrile (30 mL) was added 1,1'-thiocarbonyldiimidazole (1.405
g, 7.89 mmol). The reaction mixture was stirred at 50.degree. C.
for 18 hours. The reaction was cooled to room temperature and the
precipitate was filtered and washed with acetonitrile (2.times.50
mL). The yellow powder was dried in a vacuum oven (40.degree. C.)
for 1 hour to yield
N-(4-chlorobenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
(0.33 g, 1.12 mmol, 18.5% yield) and then used in the next step
without any further purification or characterization.
Step B:
(3-((3-(4-Chlorobenzo[d]thiazol-2-yl)thioureido)methyl)-3-hydroxy--
1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00421##
[0133] To
N-(4-chlorobenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioam-
ide (0.3 g, 1.018 mmol) in N,N-dimethylformamide (20 mL) was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (0.173 g, 1.018 mmol). The reaction mixture was stirred at
70.degree. C. for 4 hours. The reaction was concentrated and
purified via silica gel chromatography (60-100% ethyl
acetate/hexanes). The product fractions were concentrated in vacuo
to yield
(3-((3-(4-chlorobenzo[d]thiazol-2-yl)thioureido)methyl)-3-hydroxy-1-ammon-
iobicyclo[2.2.2]octan-1-yl)trihydroborate (0.25 g, 0.63 mmol, 61.9%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 12.19 (s, 1H) 9.72 (br. s., 1H) 7.93 (d, J=7.93 Hz, 1H)
7.51 (d, J=7.93 Hz, 1H) 7.26-7.32 (m, 1H) 5.32 (s, 1 H) 3.88 (dd,
J=13.73, 4.88 Hz, 1H) 3.75 (dd, J=13.73, 4.88 Hz, 1H) 2.73-2.95 (m,
6H) 2.08 (br. s., 1H) 1.96 (br. s., 1H) 1.79-1.89 (m, 1H) 1.68-1.78
(m, 1H) 1.52-1.61 (m, 1H) 1.39 (br. s., 3H).
Step C:
(2-(4-Chlorobenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole--
5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00422##
[0135] To
(3-((3-(4-chlorobenzo[d]thiazol-2-yl)thioureido)methyl)-3-hydrox-
y-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (0.23 g, 0.58
mmol) in N,N-dimethylformamide (20 mL) was added
N,N'-diisopropylcarbodiimide (0.117 mL, 0.75 mmol). The reaction
mixture was stirred at 70.degree. C. for 4 hours. The reaction was
concentrated and purified via silica gel chromatography (40-100%
ethyl acetate/hexanes). The product fractions were concentrated in
vacuo to yield
(2-(4-chlorobenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bi-
cyclo[2.2.2]octane]-1'-yl)trihydroborate (0.1 g, 0.27 mmol, 47.6%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 8.95 (s, 1H) 7.79 (d, J=7.93 Hz, 1H) 7.42 (d, J=7.63
Hz, 1H) 7.18 (t, J=7.93 Hz, 1H) 3.92 (d, J=10.38 Hz, 1H) 3.77 (d,
J=10.38 Hz, 1H) 3.26-3.38 (m, 1H) 3.19 (dd, J=15.26, 1.53 Hz, 1H)
3.01-3.11 (m, 1H) 2.81-3.00 (m, 3H) 2.31 (br. s., 1H) 2.03-2.15 (m,
1H) 1.70-1.89 (m, 3H) 1.45 (br. s., 3H).
Step D:
N-(4-Chlorobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00423##
[0137] To
(2-(4-chlorobenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazol-
e-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.08 g, 0.221
mmol) in acetone (9 mL) was added 3 M HCl (0.551 mL, 1.654 mmol).
The reaction mixture was stirred at room temperature for 4 hours.
Ethyl acetate was added and the aqueous layer was collected and
neutralized with 1N sodium hydroxide. The product was extracted
with ethyl acetate (2.times.40 mL). The organics were combined,
dried with magnesium sulfate, filtered and concentrated in vacuo to
afford
N-(4-chlorobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (0.05 g, 0.14 mmol, 65.0% yield) as a white
powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.90 (br.
s., 1H) 7.77 (dd, J=7.78, 1.07 Hz, 1H) 7.41 (dd, J=7.78, 1.07 Hz,
1H) 7.17 (t, J=7.78 Hz, 1H) 3.94 (d, J=10.07 Hz, 1H) 3.67 (d,
J=10.07 Hz, 1H) 3.00-3.11 (m, 2H) 2.76-2.88 (m, 2H) 2.68 (t, J=7.63
Hz, 2H) 2.11 (br. s., 1H) 1.91-2.00 (m, 1H) 1.47-1.67 (m, 3H). MS
(LC/MS) R.T.=2.11; [M+H].sup.+=349.1.
[0138] The enantiomers were separated using a Chiralcel OJ-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
30% methanol (0.1% DEA) in CO.sub.2. The wavelength was set at 220
nM. The separated peaks were concentrated in vacuo to yield white
powders. The first peak off the column was
(S)--N-(4-chlorobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine (0.11 g, 0.30 mmol, 34.8% yield). (7a,
S-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.90
(br. s., 1H) 7.77 (d, J=7.32 Hz, 1H) 7.41 (d, J=7.93 Hz, 1H) 7.17
(t, J=7.93 Hz, 1H) 3.94 (d, J=10.07 Hz, 1H) 3.67 (d, J=10.07 Hz,
1H) 3.00-3.11 (m, 2H) 2.76-2.90 (m, 2H) 2.68 (t, J=7.78 Hz, 2H)
2.11 (br. s., 1H) 1.91-2.01 (m, 1H) 1.47-1.68 (m, 3H). MS (LC/MS)
R.T.=2.06; [M+H].sup.+=349.1. The second peak was
(R)--N-(4-chlorobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine (0.11 g, 0.30 mmol, 35.2% yield).
(7b, R-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
8.89 (br. s., 1H) 7.77 (d, J=7.93 Hz, 1H) 7.40 (d, J=7.93 Hz, 1H)
7.16 (t, J=7.93 Hz, 1H) 3.94 (d, J=10.07 Hz, 1H) 3.67 (d, J=10.07
Hz, 1H) 2.99-3.11 (m, 2H) 2.74-2.88 (m, 2H) 2.68 (t, J=7.78 Hz, 2H)
2.11 (br. s., 1H) 1.90-2.01 (m, 1H) 1.44-1.68 (m, 3H). MS (LC/MS)
R.T.=2.07; [M+H].sup.+=349.1.
Example 8
N--(.sup.1H-Benzo[d]imidazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2-
.2]octan]-2-amine
##STR00424##
[0139] Step A:
N--(.sup.1H-Benzo[d]imidazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
##STR00425##
[0141] To .sup.1H-benzo[d]imidazol-2-amine (1.28 g, 9.61 mmol) in
acetonitrile (30 mL) was added 1,1'-thiocarbonyldiimidazole (2.227
g, 12.5 mmol). The reaction mixture was stirred at 50.degree. C.
for 18 hours. The reaction was cooled to room temperature and the
precipitate was filtered and washed with acetonitrile (2.times.50
mL). The yellow powder was dried in a vacuum oven (40.degree. C.)
for 1 hour to yield
N-(1H-benzo[d]imidazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
(1.8 g, 7.4 mmol, 77% yield) and then used in the next step without
any further purification or characterization.
Step B:
(3-((3-.sup.1H-Benzo[d]-imidazol-2-ylthioureido)methyl)-3-hydroxy--
1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00426##
[0143] To
N--(.sup.1H-benzo[d]imidazol-2-yl)-.sup.1H-imidazole-1-carbothio-
amide (1.07 g, 4.4 mmol) in N,N-dimethylformamide (20 mL) was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (0.748 g, 4.4 mmol). The reaction mixture was stirred at
70.degree. C. for 4 hours. The reaction was concentrated and
purified via silica gel chromatography (60-100% ethyl
acetate/hexane). The product fractions were concentrated in vacuo
to yield
(3-((3-.sup.1H-benzo[d]imidazol-2-ylthioureido)methyl)-3-hydroxy-1-ammoni-
obicyclo[2.2.2]octan-1-yl)trihydroborate (1.28 g, 3.71 mmol, 84%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 11.41 (s, 1H), 11.16 (s, 1H), 7.43 (d, J=3.05 Hz, 3H),
7.13 (ddd, J=9.46, 3.81, 3.51 Hz, 3H), 5.36 (s, 1H), 4.01-4.07 (m,
3H), 3.79 (dd, J=13.28, 4.12 Hz, 1 H), 2.83-2.92 (m, 6H), 2.71 (d,
J=14.04 Hz, 2H), 2.04-2.13 (m, 2H), 1.89-1.94 (m, 3H), 1.74 (td,
J=9.46, 5.49 Hz, 2H), 1.51-1.59 (m, 2H), 1.38 (s, 2H), 1.31 (s,
1H). LC/MS confirmed product as loss of BH.sub.3 in the LC/MS
conditions: retention time 2.75 (M+1-BH.sub.3=332.2).
Step C:
(2-(.sup.1H-Benzo[d]-imidazol-2-ylamino)-4H-1'-ammoniospiro-[oxazo-
le-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00427##
[0145] To
(3-((3-.sup.1H-benzo[d]imidazol-2-ylthioureido)methyl)-3-hydroxy-
-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (1.0 g, 2.9 mmol)
in N,N-dimethylformamide (20 mL) was added
N,N'-diisopropylcarbodiimide (0.587 mL, 3.77 mmol). The reaction
mixture was stirred at 70.degree. C. for 4 hours. The reaction was
concentrated and purified via silica gel chromatography (40-100%
ethyl acetate/hexane). The product fractions were concentrated in
vacuo to yield
(2-(.sup.1H-benzo[d]imidazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-b-
icyclo[2.2.2]octane]-1'-yl)trihydroborate (0.81 g, 2.6 mmol, 90%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 7.29 (s, 1H), 6.99-7.05 (m, 2H), 3.90 (t, J=9.77 Hz,
1H), 3.75 (d, J=10.38 Hz, 1H), 3.26-3.35 (m, 1H), 3.13 (dd,
J=14.95, 1.53 Hz, 1H), 2.98-3.06 (m, 1H), 2.84-2.92 (m, 3H), 2.22
(s, 1H), 1.98-2.05 (m, 1H), 1.72-1.82 (m, 3H), 1.45 (s, 1H). LC/MS
confirmed product as loss of BH.sub.3 in the LC/MS conditions:
retention time 2.29 (M+1-BH.sub.3=298.2).
Step D:
N--(.sup.1H-Benzo[d]imidazol-2-yl)-4H-1'-azaspiro-[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine
##STR00428##
[0147] To
(2-(.sup.1H-benzo[d]imidazol-2-ylamino)-4H-1'-ammoniospiro[oxazo-
le-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.77 g, 2.5
mmol) in acetone (9 mL) was added 3 M HCl (6.2 mL, 18.6 mmol). The
reaction was stirred at room temperature for 4 hours. Ethyl acetate
was added and the aqueous layer was collected and neutralized with
1N sodium hydroxide. The product was extracted with ethyl acetate
(2.times.40 mL). The organics were combined, dried with magnesium
sulfate, filtered and concentrated in vacuo to afford
N--(.sup.1H-benzo[d]imidazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.-
2.2]octan]-2-amine (0.5 g, 1.68 mmol, 68% yield) as a white powder.
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 11.45 (s, 1H), 9.20
(s, 1H), 7.32 (s, 1H), 7.00 (dd, J=5.65, 2.90 Hz, 4H), 3.91 (d,
J=10.07 Hz, 2H), 3.64 (d, J=10.07 Hz, 2H), 2.98-3.05 (m, 4H),
2.73-2.82 (m, 4H), 2.67 (t, J=7.63 Hz, 4H), 2.03 (d, J=2.75 Hz,
2H), 1.85-1.92 (m, 2H), 1.54-1.63 (m, 4H), 1.44-1.51 (m, 2 H). MS
(LC/MS) R.T.=1.30; [M+H].sup.+=298.2.
[0148] The enantiomers were separated using a Chiralpak AS-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
30% methanol (0.1% DEA) in CO.sub.2 and UV monitored at 330 nm. The
separated peaks were concentrated in vacuo to yield white powders.
The first peak off the column yielded 0.11 g, 0.36 mmol, 33.0%.
(8a; R-isomer): M.P. 255.degree. C. (dec). .sup.1H NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 11.43 (br. s., 1 H) 9.16 (br. s., 1H)
7.14-7.53 (m, 2H) 6.82-7.09 (m, 2H) 3.91 (d, J=9.77 Hz, 1H) 3.64
(d, J=10.07 Hz, 1H) 2.96-3.09 (m, 2H) 2.71-2.87 (m, 2H) 2.62-2.73
(m, 2H) 2.01-2.08 (m, 1H) 1.81-1.97 (m, 1H) 1.54-1.66 (m, 2H)
1.40-1.53 (m, 1 H). MS (LC/MS) R.T.=1.26; [M+H].sup.+=298.2. The
second peak yielded 0.11 g, 0.36 mmol, 33.0%. (8b; S-isomer):
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 11.43 (br. s., 1H)
9.20 (br. s., 1H) 7.10-7.60 (m, 2H) 6.83-7.11 (m, 2H) 3.91 (d,
J=9.77 Hz, 1H) 3.64 (d, J=9.77 Hz, 1H) 2.94-3.16 (m, 2H) 2.71-2.86
(m, 2H) 2.59-2.72 (m, 2H) 1.97-2.09 (m, 1H) 1.81-1.95 (m, 1H)
1.53-1.71 (m, 2H) 1.41-1.53 (m, 1H). MS (LC/MS) R.T.=1.28;
[M+H].sup.+=298.2.
Example 9
N-(6-Chlorobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00429##
[0149] Step A:
N-(6-Chlorobenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
##STR00430##
[0151] To 6-chlorobenzo[d]thiazol-2-amine (1.14 g, 6.17 mmol) in
acetonitrile (30 mL) was added 1,1'-thiocarbonyldiimidazole (1.43
g, 8 mmol). The reaction was stirred at 50.degree. C. for 18 hours.
The reaction was cooled to room temperature and the precipitate was
filtered and washed with acetonitrile (2.times.50 mL). The yellow
powder was dried in a vacuum oven (40.degree. C.) for 1 hour to
yield
N-(6-chlorobenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
(1.16 g, 3.9 mmol, 64% yield) and then used in the next step
without any further purification or characterization.
Step B:
(3-((3-(6-Chlorobenzo[d]thiazol-2-yl)thioureido)methyl)-3-hydroxy--
1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00431##
[0153] To
N-(6-chlorobenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioam-
ide (0.86 g, 2.9 mmol) in N,N-dimethylformamide (20 mL) was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (0.5 g, 2.9 mmol). The reaction was stirred at 70.degree. C. for
4 hours. The reaction was concentrated and purified via silica gel
chromatography (60-100% ethyl acetate/hexanes). The product
fractions were concentrated in vacuo to yield
(3-((3-(6-chlorobenzo[d]thiazol-2-yl)thioureido)methyl)-3-hydroxy-1-ammon-
iobicyclo[2.2.2]octan-1-yl)trihydroborate (0.4 g, 1.01 mmol, 34.6%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 11.92 (br. s., 1H) 9.89 (br. s., 1H) 8.08 (br. s., 1H)
7.62 (br. s., 1H) 7.40-7.50 (m, 1H) 5.40 (br. s., 1H) 3.88 (d,
J=10.20 Hz, 1H) 3.76 (d, J=10.20 Hz, 1H) 2.67-3.02 (m, 6H) 2.08
(br. s., 1H) 1.80-1.95 (m, 2H) 1.73 (br. s., 1H) 1.01-1.63 (m, 4H).
MS (LC/MS) R.T.=3.87; [M+H].sup.+=395.1.
Step C:
(2-(6-Chlorobenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole--
5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00432##
[0155] To
(3-((3-(6-chlorobenzo[d]thiazol-2-yl)thioureido)methyl)-3-hydrox-
y-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (0.37 g, 0.93
mmol) in N,N-dimethylformamide (20 mL) was added
N,N'-diisopropylcarbodiimide (0.19 mL, 1.2 mmol). The reaction was
stirred at 70.degree. C. for 4 hours. The reaction was concentrated
and purified via silica gel chromatography (40-100% ethyl
acetate/hexane). The product fractions were concentrated in vacuo
to yield
(2-(6-chlorobenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bi-
cyclo[2.2.2]octane]-1'-yl)trihydroborate (0.12 g, 0.33 mmol, 35.5%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 9.10 (br. s., 1H) 7.95 (d, J=2.14 Hz, 1H) 7.60 (d,
J=8.55 Hz, 1H) 7.36 (dd, J=8.55, 2.14 Hz, 1H) 3.88 (d, J=10.38 Hz,
1H) 3.76 (d, J=10.38 Hz, 1H) 3.26-3.37 (m, 1H) 3.17 (dd, J=14.95,
1.83 Hz, 1H) 3.00-3.11 (m, 1H) 2.80-2.97 (m, 3H) 2.28 (br. s., 1H)
2.00-2.11 (m, 1H) 1.69-1.88 (m, 3 H) 1.46 (br. s., 3H). LC/MS:
retention time 2.94 (M+1-BH.sub.3=349.1).
Step D:
N-(6-Chlorobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00433##
[0157] To
(2-(6-chlorobenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazol-
e-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.1 g, 0.28
mmol) in acetone (9 mL) was added 3 M HCl (0.69 mL, 2.07 mmol). The
reaction was stirred at room temperature for 4 hours. Ethyl acetate
was added and the aqueous layer was collected and neutralized with
1N sodium hydroxide. The product was extracted with ethyl acetate
(2.times.40 mL). The organics were combined, dried with magnesium
sulfate, filtered and concentrated in vacuo to afford
N-(6-chlorobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (0.06 g, 0.17 mmol, 62.4% yield) as a white
powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.03 (br.
s., 1H) 7.93 (d, J=2.14 Hz, 1H) 7.58 (d, J=8.55 Hz, 1H) 7.35 (dd,
J=8.55, 2.14 Hz, 1 H) 3.90 (d, J=10.07 Hz, 1H) 3.65 (d, J=10.07 Hz,
1H) 2.99-3.11 (m, 2H) 2.73-2.89 (m, 2H) 2.67 (t, J=7.63 Hz, 2H)
2.08 (br. s., 1H) 1.82-1.99 (m, 1H) 1.43-1.67 (m, 3H). MS (LC/MS)
R.T.=2.07; [M+H].sup.+=349.1.
[0158] The enantiomers were separated using a Chiralpak AS-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
30% methanol (0.1% DEA) in CO.sub.2 and UV monitored at 220 nm. The
separated peaks were concentrated in vacuo to yield white powders.
The first peak off the column yielded 0.034 g, 0.10 mmol, 36.2%.
(9a; S-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
9.02 (br. s., 1H) 7.93 (d, J=2.14 Hz, 1H) 7.58 (d, J=8.55 Hz, 1H)
7.35 (dd, J=8.55, 2.14 Hz, 1H) 3.90 (d, J=10.07 Hz, 1 H) 3.65 (d,
J=10.07 Hz, 1H) 2.97-3.11 (m, 2H) 2.73-2.89 (m, 2H) 2.67 (t, J=7.48
Hz, 2H) 2.08 (br. s., 1H) 1.93 (br. s., 1H) 1.43-1.69 (m, 3H). MS
(LC/MS) R.T.=2.05; [M+H].sup.+=349.1. Optical
rotation=+4.00.degree.. The second peak yielded 0.037 g, 0.10 mmol,
39.4%. (9b; R-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 9.02 (br. s., 1H) 7.93 (d, J=2.14 Hz, 1H) 7.58 (d, J=8.55 Hz,
1H) 7.35 (dd, J=8.55, 2.44 Hz, 1H) 3.90 (d, J=10.07 Hz, 1H) 3.65
(d, J=10.07 Hz, 1H) 2.93-3.13 (m, 2H) 2.72-2.91 (m, 2H) 2.62-2.73
(m, 2H) 2.08 (br. s., 1H) 1.93 (d, J=1.22 Hz, 1H) 1.44-1.68 (m,
3H). MS (LC/MS) R.T.=2.04; [M+H].sup.+=349.1. Optical
rotation=-3.74.degree..
Example 10
N-(1-Methyl-.sup.1H-benzo[d]imidazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00434##
[0159] Step A:
N-(1-Methyl-.sup.1H-benzo[d]imidazol-2-yl)-.sup.1H-imidazole-1-carbothioa-
mide
##STR00435##
[0161] To 1-methyl-.sup.1H-benzo[d]imidazol-2-amine (1.28 g, 8.7
mmol) in acetonitrile (30 mL) was added
1,1'-thiocarbonyldiimidazole (2.015 g, 11.31 mmol). The reaction
was stirred at 50.degree. C. for 18 hours. The reaction was cooled
to room temperature and the precipitate was filtered and washed
with acetonitrile (2.times.50 mL). The yellow powder was dried in a
vacuum oven (40.degree. C.) for 1 hour to yield
N-(1-methyl-.sup.1H-benzo[d]imidazol-2-yl)-.sup.1H-imidazole-1-carbothioa-
mide (1.6 g, 6.22 mmol, 71.5% yield) and then used in the next step
without any further purification or characterization.
Step B:
(3-Hydroxy-3-((3-(1-methyl-.sup.1H-benzo[d]imidazol-2-yl)thioureid-
o)methyl)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00436##
[0163] To
N-(1-methyl-.sup.1H-benzo[d]imidazol-2-yl)-.sup.1H-imidazole-1-c-
arbothioamide (1.04 g, 4.04 mmol) in N,N-dimethylformamide (20 mL)
was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihyd-
roborate (0.687 g, 4.04 mmol). The reaction was stirred at
70.degree. C. for 4 hours. The reaction was concentrated and
purified via silica gel chromatography (60-100% ethyl
acetate/hexane). The product fractions were concentrated in vacuo
to yield
(3-hydroxy-3-((3-(1-methyl-.sup.1H-benzo[d]imidazol-2-yl)thioureido)methy-
l)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (1.28 g, 3.56
mmol, 88% yield) as a white powder. LC/MS confirmed product as loss
of BH.sub.3 in the LC/MS conditions: retention time 3.01
(M+1-BH.sub.3=346.2).
Step C:
(2-(1-Methyl-.sup.1H-benzo[d]imidazol-2-ylamino)-4H-1'-ammoniospir-
o[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00437##
[0165] To
(3-hydroxy-3-((3-(1-methyl-.sup.1H-benzo[d]imidazol-2-yl)thioure-
ido)methyl)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (1.19
g, 3.31 mmol) in N,N-dimethylformamide (20 mL) was added
N,N'-diisopropylcarbodiimide (1.55 mL, 9.9 mmol). The reaction was
stirred at 70.degree. C. for 4 hours. The reaction was concentrated
and purified via silica gel chromatography (40-100% ethyl
acetate/hexane). The product fractions were concentrated in vacuo
to yield
(2-(1-methyl-.sup.1H-benzo[d]imidazol-2-ylamino)-4H-1'-ammoniospiro[oxazo-
le-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.94 g, 2.9
mmol, 87% yield) as a white powder. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.36 (br. s., 1H) 7.34-7.44 (m, 1H)
7.26-7.34 (m, 1H) 7.00-7.12 (m, 2H) 3.90 (d, J=10.32 Hz, 1H) 3.77
(d, J=10.32 Hz, 1H) 3.57 (s, 3H) 3.28 (dd, J=14.86, 2.27 Hz, 1H)
3.13 (dd, J=14.86, 1.51 Hz, 1H) 2.95-3.08 (m, 1H) 2.75-2.95 (m, 3H)
2.22 (br. s., 1H) 1.96-2.11 (m, 1H) 1.67-1.89 (m, 3H) 1.43 (br. s.,
3H). LC/MS: retention time 2.37 (M+1-BH.sub.3=312.2).
Step D:
N-(1-Methyl-.sup.1H-benzo[d]imidazol-2-yl)-4H-1'-azaspiro[oxazole--
5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00438##
[0167] To
(2-(1-methyl-.sup.1H-benzo[d]imidazol-2-ylamino)-4H-1'-ammoniosp-
iro[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.92
g, 2.8 mmol) in acetone (9 mL) was added 3 M HCl (7.1 mL, 21.2
mmol). The reaction was stirred at room temperature for 4 hours.
Ethyl acetate was added and the aqueous layer was collected and
neutralized with 1N sodium hydroxide. The product was extracted
with ethyl acetate (2.times.40 mL). The organics were combined,
dried with magnesium sulfate, filtered, and concentrated in vacuo
to afford
N-(1-methyl-.sup.1H-benzo[d]imidazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine (0.84 g, 2.7 mmol, 95% yield) as a white
powder. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.24-9.39
(m, 1H) 7.33-7.42 (m, 1H) 7.25-7.33 (m, 1H) 6.99-7.10 (m, 2 H) 3.92
(d, J=10.07 Hz, 1H) 3.66 (d, J=10.07 Hz, 1H) 3.57 (s, 3H) 2.97-3.08
(m, 2H) 2.78 (t, J=7.81 Hz, 2H) 2.67 (t, J=7.81 Hz, 2H) 2.01-2.09
(m, 1H) 1.80-1.96 (m, 1H) 1.40-1.66 (m, 3H). MS (LC/MS) R.T.=1.49;
[M+H].sup.+=312.2.
[0168] The enantiomers were separated using a Chiralcel OJ-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
22% methanol (0.1% DEA) in CO.sub.2 and UV monitored at 300 nm. The
separated peaks were concentrated in vacuo to yield white powders.
The first peak off the column yielded 0.065 g, 0.205 mmol, 39.8%.
(10a, R-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
9.31 (br. s., 1H) 7.35-7.45 (m, 1 H) 7.26-7.34 (m, 1H) 7.00-7.15
(m, 2H) 3.93 (d, J=10.10 Hz, 1H) 3.67 (d, J=10.10 Hz, 1H) 3.59 (s,
3H) 2.93-3.13 (m, 2H) 2.74-2.89 (m, 2H) 2.61-2.74 (m, 2H) 2.05 (br.
s., 1H) 1.91 (br. s., 1H) 1.38-1.68 (m, 3H). MS (LC/MS) R.T.=1.37;
[M+H].sup.+=312.2. Optical rotation=-16.02.degree.. The second peak
yielded 0.06 g, 0.19 mmol, 36.8%. (10b; S-isomer): .sup.1H NMR (500
MHz, DMSO-d.sub.6) .delta. ppm 9.31 (br. s., 1H) 7.33-7.43 (m, 1H)
7.25-7.33 (m, 1H) 6.93-7.11 (m, 2H) 3.93 (dd, J=9.92, 3.20 Hz, 1H)
3.67 (dd, J=9.92, 3.20 Hz, 1H) 3.58 (s, 3H) 2.94-3.13 (m, 2 H)
2.74-2.85 (m, 2H) 2.59-2.72 (m, 2H) 2.04 (br. s., 1H) 1.90 (br. s.,
1H) 1.37-1.70 (m, 3H). MS (LC/MS) R.T.=1.37; [M+H].sup.+=312.2.
Optical rotation=+35.99.degree..
Example 11
N-(6-Ethoxybenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00439##
[0169] Step A:
N-(6-Ethoxybenzo[d]thiazol-2-yl)-1H-imidazole-1-carbothioamide
##STR00440##
[0171] In a vial was placed 6-ethoxybenzo[d]thiazol-2-amine (1.5 g,
7.72 mmol) and di(.sup.1H-imidazol-1-yl)methanethione (1.789 g,
10.04 mmol) in acetonitrile (15 mL). The reaction was heated to
80.degree. C. overnight. The reaction mixture was filtered and the
precipitate was collected to afford 2.4 grams (7.88 mmol, 102%) of
rust-colored solids.
Step B:
(3-((3-(6-Ethoxybenzo[d]thiazol-2-yl)thioureido)methyl)-3-hydroxy--
1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00441##
[0173] In a vial was placed
N-(6-ethoxybenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
(2.4 g, 7.88 mmol) and
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (1.341 g, 7.88 mmol) in N,N-dimethylformamide (8 mL). The
reaction was heated to 80.degree. C. After 2 hours the reaction was
poured into water and chloroform and the organic was extracted and
concentrated to a red oil. This material was used in the next
reaction without any further purification or characterization.
Step C:
(2-(6-Ethoxybenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro-[oxazole-
-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00442##
[0175] In a flask was placed
(3-((3-(6-ethoxybenzo[d]thiazol-2-yl)thioureido)-methyl)-3-hydroxy-1-ammo-
niobicyclo[2.2.2]octan-1-yl)trihydroborate (3.2 g, 7.9 mmol) and
N,N'-diisopropylcarbodiimide (4.3 mL, 27.6 mmol) in
N,N-dimethylformamide (10 mL). The reaction was heated to
70.degree. C. for 2 hours and then poured into water and
chloroform. The organic was collected and concentrated to a
residue. The residue was triturated in ether and the precipitate
was collected via vacuum filtration to yield 1.11 grams (2.98 mmol,
37.8%) of gray powder.
Step D:
N-(6-Ethoxybenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00443##
[0177] In a vial was placed
(2-(6-ethoxybenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bi-
cyclo[2.2.2]octane]-1'-yl)trihydroborate (500 mg, 1.34 mmol) and
HCl (8.06 mL, 24.17 mmol) in acetone (10 mL). The reaction was
monitored by HPLC. After 2 hours, the reaction was complete by
LC/MS. The reaction was poured into water and chloroform, and the
organic layer was set aside. The aqueous layer was neutralized and
extracted with chloroform (2.times.). The second chloroform
fraction was concentrated to a white residue. The solid was
triturated in ether and the precipitate collected to yield 314.4 mg
(0.877 mmol, 65.3%) of the desired material. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 8.89 (1H, br. s.), 7.50 (1H, d, J=8.81
Hz), 7.38 (1H, d, J=2.52 Hz), 6.91 (1H, dd, J=8.81, 2.52 Hz), 4.03
(2H, q, J=7.05 Hz), 3.87 (1H, d, J=10.07 Hz), 3.62 (1H, d, J=10.07
Hz), 3.02 (2H, s), 2.78 (2H, t, J=7.81 Hz), 2.66 (2H, t, J=7.68
Hz), 2.05 (1H, br. s.), 1.91 (1H, br. s.), 1.42-1.67 (3H, m), 1.33
(3H, t, J=7.05 Hz). MS (LC/MS) R.T.=1.60; [M+H].sup.+=359.0.
[0178] The enantiomers were separated using a Chiralpak AS-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
30% methanol (0.1% DEA) in CO.sub.2 and UV monitored at 300 nm. The
separated peaks were concentrated in vacuo to yield white powders.
The first peak off the column yielded 62.4 mg, 0.17 mmol, 31.2%.
(11a; S-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
8.87 (1H, br. s.), 7.49 (1H, d, J=8.55 Hz), 7.38 (1H, d, J=2.44
Hz), 6.91 (1H, dd, J=8.85, 2.44 Hz), 4.03 (2H, q, J=7.02 Hz), 3.87
(1H, d, J=10.07 Hz), 3.61 (1H, d, J=10.07 Hz), 3.02 (2H, s),
2.72-2.85 (2H, m), 2.66 (2H, t, J=7.63 Hz), 2.05 (1H, br. s.), 1.91
(1H, br. s.), 1.43-1.64 (3H, m), 1.33 (3H, t, J=7.02 Hz). MS
(LC/MS) R.T.=1.81; [M+H].sup.+=359.1. The second peak yielded 58.9
mg, 0.164 mmol, 29.5%. (11b; R-isomer): .sup.1H NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 8.86 (1H, br. s.), 7.48 (1H, d, J=8.85
Hz), 7.37 (1H, d, J=2.44 Hz), 6.91 (1H, d, J=2.75 Hz), 4.02 (2H, q,
J=7.02 Hz), 3.86 (1H, d, J=9.77 Hz), 3.61 (1H, d, J=9.77 Hz), 3.01
(2H, s), 2.72-2.85 (2H, m), 2.62-2.69 (2H, m), 2.04 (1H, d, J=2.44
Hz), 1.90 (1H, d, J=4.27 Hz), 1.42-1.63 (3H, m), 1.29-1.35 (3H, m).
MS (LC/MS) R.T.=1.52; [M+H].sup.+=359.1.
Example 12
N-(6-Methylbenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00444##
[0179] Step A:
N-(6-Methylbenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
##STR00445##
[0181] To 6-methylbenzo[d]thiazol-2-amine (1. g, 6.2 mmol) in
acetonitrile (30 mL) was added 1,1'-thiocarbonyldiimidazole (1.44
g, 8.1 mmol). The reaction was stirred at 50.degree. C. for 18
hours. The reaction was cooled to room temperature and the
precipitate was filtered and washed with acetonitrile (2.times.50
mL). The yellow powder was dried in a vacuum oven (40.degree. C.)
for 1 hour to yield
N-(6-methylbenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioamide
(1.06 g, 3.86 mmol, 62% yield) and then used in the next step
without any further purification or characterization.
Step B:
(3-Hydroxy-3-((3-(6-methylbenzo[d]thiazol-2-yl)thioureido)methyl)--
1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00446##
[0183] To
N-(6-methylbenzo[d]thiazol-2-yl)-.sup.1H-imidazole-1-carbothioam-
ide (0.96 g, 3.5 mmol) in N,N-dimethylformamide (20 mL) was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (0.595 g, 3.5 mmol). The reaction was stirred at 70.degree. C.
for 4 hours. The reaction mixture was concentrated and purified via
silica gel chromatography (40-100% ethyl acetate/hexanes). The
product fractions were concentrated in vacuo to yield
(3-hydroxy-3-((3-(6-methylbenzo[d]thiazol-2-yl)thioureido)methyl)-1-ammon-
iobicyclo[2.2.2]octan-1-yl)trihydroborate (0.88 g, 2.338 mmol,
66.8% yield) as a white powder. MS (LC/MS) R.T.=3.71;
[M+H].sup.+=375.2.
Step C:
(2-(6-Methylbenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole--
5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00447##
[0185] To
(3-hydroxy-3-((3-(6-methylbenzo[d]thiazol-2-yl)thioureido)methyl-
)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (0.86 g, 2.285
mmol) in N,N-dimethylformamide (20 mL) was added
N,N'-diisopropylcarbodiimide (0.288 g, 2.285 mmol). The reaction
was stirred at 70.degree. C. for 4 hours. The reaction was
concentrated and ethyl acetate was added. The precipitate was
filtered and washed with additional ethyl acetate. The powder was
dried in a vacuum oven (70.degree. C.) to yield
(2-(6-methylbenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bi-
cyclo[2.2.2]octane]-1'-yl)trihydroborate (0.65 g, 1.899 mmol, 83%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 9.03 (1H, br. s.), 7.59 (1H, s), 7.51 (1H, d, J=8.24
Hz), 7.15 (1H, d, J=8.24 Hz), 3.86 (1H, d, J=10.38 Hz), 3.74 (1H,
d, J=10.38 Hz), 3.29 (1H, dd, J=15.26, 1.83 Hz), 3.14 (1H, d,
J=15.26 Hz), 2.99-3.09 (1H, m), 2.80-2.95 (3H, m), 2.36 (3H, s),
2.25 (1H, br. s.), 2.03 (1H, t, J=10.22 Hz), 1.70-1.85 (3H, m). MS
(LC/MS) R.T.=2.78; [M+H].sup.+=343.2.
Step D:
N-(6-Methylbenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00448##
[0187] To
(2-(6-methylbenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazol-
e-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.24 g, 0.70
mmol) in acetone (9 mL) was added 3 M HCl (1.753 mL, 5.26 mmol).
The reaction mixture was stirred at room temperature for 4 hours.
Ethyl acetate was added and the aqueous layer was collected and
neutralized with 1N sodium hydroxide. The product was extracted
with ethyl acetate (2.times.40 mL). The organics were combined,
dried with magnesium sulfate, filtered and concentrated in vacuo to
afford
N-(6-methylbenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (0.19 g, 0.58 mmol, 83% yield) as a white powder.
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.96 (1H, br. s.),
7.57 (1H, s), 7.49 (1H, d, J=8.24 Hz), 7.13 (1H, d, J=8.24 Hz),
3.88 (1H, d, J=10.07 Hz), 3.63 (1H, d, J=9.77 Hz), 2.98-3.04 (2H,
m), 2.72-2.85 (2H, m), 2.66 (2H, t, J=7.63 Hz), 2.36 (3H, s), 2.05
(1H, d, J=2.14 Hz), 1.91 (1H, br. s.), 1.53-1.64 (2H, m), 1.42-1.53
(1H, m). MS (LC/MS) R.T.=1.79; [M+H].sup.+=329.2.
[0188] The enantiomers were separated using a Chiralcel OJ-H
(4.6.times.25 cm, 5 .mu.m) column with 30% methanol (0.1% DEA) in
CO.sub.2 and UV monitored at 300 nm. The separated peaks were
concentrated in vacuo to yield white powders. The first peak off
the column yielded 0.11 g, 0.34 mmol, 55%. (12a; R-isomer): .sup.1H
NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.95 (s, 1H), 7.57 (s, 2H),
7.48 (d, J=8.24 Hz, 2H), 7.13 (d, J=8.24 Hz, 2H), 3.89 (d, J=10.38
Hz, 2H), 3.63 (d, J=10.38 Hz, 2H), 2.99-3.06 (m, 4H), 2.75-2.84 (m,
4H), 2.67 (t, J=7.63 Hz, 4H), 2.37 (s, 7H), 2.06 (s, 2H), 1.92 (s,
2H), 1.55-1.64 (m, 4H), 1.49 (dd, J=9.77, 2.44 Hz, 2H). MS (LC/MS)
R.T.=1.80; [M+H].sup.+=329.2. Optical rotation=-4.52. The second
peak yielded 0.11 g, 0.34 mmol, 55%. (12b; S-isomer): .sup.1H NMR
(500 MHz, DMSO-d.sub.6) .delta. ppm 8.96 (s, 1H), 7.58 (s, 1H),
7.49 (d, J=8.24 Hz, 1H), 7.14 (d, J=8.24 Hz, 1H), 3.89 (d, J=10.07
Hz, 1H), 3.63 (d, J=10.07 Hz, 1H), 3.03 (d, J=2.44 Hz, 2H),
2.75-2.84 (m, 2H), 2.67 (t, J=7.78 Hz, 2H), 2.37 (s, 3H), 2.06 (s,
1H), 1.92 (s, 1H), 1.55-1.64 (m, 2H), 1.49 (dd, J=9.77, 2.75 Hz,
1H). MS (LC/MS) R.T.=1.80; [M+H].sup.+=329.2. Optical
rotation=+10.08.
Example 13
2-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)benzo[d]thi-
azol-6-ol
##STR00449##
[0189] Step A:
N-(6-(tert-Butyldimethylsilyloxy)benzo[d]thiazol-2-yl)-1H-imidazole-1-car-
bothioamide
##STR00450##
[0191] To 6-(tert-butyldimethylsilyloxy)benzo[d]thiazol-2-amine
(prepared as described in WO 2007/086800 p. 102) (3.1 g, 11.05
mmol) in acetonitrile (30 mL) was added thiocarbonyl diimidazole
(2.56 g, 14.37 mmol). The reaction was heated to 70.degree. C.
overnight. The reaction was cooled to room temperature and the
precipitate was filtered and washed with acetonitrile to afford a
yellow solid. The product,
N-(6-(tert-butyldimethylsilyloxy)benzo[d]thiazol-2-yl)-1H-imidazole-1-car-
bothioamide (3.85 g, 9.86 mmol, 89% yield), was taken directly to
the next step without any further purification. MS (LC/MS)
R.T.=2.56; [M+H].sup.+=388.9.
Step B:
(3-((3-(6-(tert-Butyldimethylsilyloxy)benzo[d]thiazol-2-yl)thioure-
ido)methyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00451##
[0193] To
N-(6-(tert-butyldimethylsilyloxy)benzo[d]thiazol-2-yl)-1H-imidaz-
ole-1-carbothioamide (3.85 g, 9.86 mmol) in N,N-dimethylformamide
(40 mL) was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)tr-
ihydroborate (1.68 g, 9.86 mmol). The reaction was heated at
80.degree. C. for 2 hours. The reaction was cooled and then poured
into a mixture of chloroform and water. The organic layer was
collected and concentrated in vacuo to afford
(3-((3-(6-(tert-butyldimethylsilyloxy)benzo[d]thiazol-2-yl)thioureido)met-
hyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
(6.2 g) as a yellow oil.
Step C:
(2-(6-(tert-Butyldimethylsilyloxy)benzo[d]thiazol-2-ylamino)-4H-1'-
-ammoniospiro[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00452##
[0195] To
(3-((3-(6-(tert-butyldimethylsilyloxy)benzo[d]thiazol-2-yl)thiou-
reido)methyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
(4.9 g, 9.9 mmol) in N,N-dimethylformamide (10 mL) was added
1,3-diisopropylcarbodiimide (5.4 mL, 34.5 mmol). The reaction was
heated to 80.degree. C. and monitored by LC/MS. The reaction was
cooled and then poured into a mixture of chloroform and water. The
organic layer was collected and concentrated in vacuo. The
remaining residue was triturated in ether. The precipitate was
collected to afford
(2-(6-(tert-butyldimethylsilyloxy)benzo[d]thiazol-2-ylamino)-4H-1'-ammoni-
ospiro[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
(3.28 g, 7.15 mmol, 72.6% yield). MS (LC/MS) R.T.=3.60;
[M+H-BH.sub.3].sup.+=445.2.
Step D:
2-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)ben-
zo[d]thiazol-6-ol
##STR00453##
[0197] To
(2-(6-(tert-butyldimethylsilyloxy)benzo[d]thiazol-2-ylamino)-4H--
1'-ammoniospiro[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
(3.2 g, 6.98 mmol) in acetone (10 mL) was added HCl (8.14 mL, 24.43
mmol). The reaction was stirred at room temperature for 3 hours.
The mixture was poured into water and neutralized with saturated
sodium bicarbonate. The aqueous layer was then extracted with
chloroform. The organic layer was collected and concentrated in
vacuo. The remaining residue was triturated in ether to afford
racemic
2-(4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)benzo[d]th-
iazol-6-ol (958 mg, 2.90 mmol, 41.5% yield) as a white powder.
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.36 (1H, br. s.),
8.85 (1H, br. s.), 7.41 (1H, d, J=8.85 Hz), 7.12 (1H, d, J=2.44
Hz), 6.78 (1H, dd, J=8.85, 2.44 Hz), 3.86 (1H, d, J=10.07 Hz), 3.61
(1H, d, J=10.07 Hz), 2.96-3.06 (2H, m), 2.71-2.85 (2H, m), 2.66
(2H, t, J=7.78 Hz), 2.04 (1H, br. s.), 1.90 (1H, br. s.), 1.43-1.64
(3H, m). MS (LC/MS) R.T.=1.03; [M+H].sup.+=331.29.
[0198] The enantiomers were separated using a Chiralpak AD-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
30% methanol (0.1% DEA) in CO.sub.2. The wavelength was set at 300
nM. The separated peaks were concentrated in vacuo to yield white
powders. The first peak off the column was
(S)-2-(4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)benzo[-
d]thiazol-6-ol (395.2 mg, 1.19 mmol, 41.4% yield). (13a, S-isomer):
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 9.36 (1H, br. s.),
8.84 (1H, br. s.), 7.41 (1H, d, J=8.55 Hz), 7.12 (1H, d, J=2.44
Hz), 6.78 (1H, dd, J=8.55, 2.44 Hz), 3.86 (1H, d, J=9.77 Hz), 3.61
(1H, d, J=10.07 Hz), 2.96-3.05 (2H, m), 2.72-2.84 (2H, m), 2.65
(2H, t, J=7.63 Hz), 2.04 (1H, br. s.), 1.90 (1H, br. s.), 1.52-1.64
(2H, m), 1.43-1.52 (1H, m). MS (LC/MS) R.T.=1.30;
[M+H].sup.+=331.4. The second peak was
(R)-2-(4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)benzo[-
d]thiazol-6-ol (375 mg, 1.14 mmol, 39.3% yield). (13b, R-isomer):
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 7.40 (1H, d, J=8.55
Hz), 7.11 (1H, d, J=2.44 Hz), 6.77 (1H, dd, J=8.55, 2.44 Hz), 3.86
(1H, d, J=10.07 Hz), 3.60 (1H, d, J=10.07 Hz), 3.01 (2H, s),
2.73-2.84 (2H, m), 2.65 (2H, t, J=7.78 Hz), 2.04 (1H, br. s.), 1.90
(1H, br. s.), 1.54-1.62 (2H, m), 1.43-1.52 (1H, m). MS (LC/MS)
R.T.=1.43; [M+H].sup.+=331.4.
Example 14
N-(4,5,6,7-Tetrahydrobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00454##
[0199] Step A:
N-(4,5,6,7-Tetrahydrobenzo[d]thiazol-2-yl)-1H-imidazole-1-carbothioamide
##STR00455##
[0201] To 4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (0.5 g, 3.24
mmol) in acetonitrile (20 mL) was added
di(1H-imidazol-1-yl)methanethione (0.58 g, 3.24 mmol). The reaction
was stirred at 50.degree. C. for 4 hours. The reaction was cooled
and concentrated to yield crude product. The crude material was
purified via flash chromatography (50-100% ethyl acetate-hexane) to
yield
N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-imidazole-1-carbothioamide
(0.60 g, 2.27 mmol, 70.0% yield) as a white powder.
Step B:
(3-Hydroxy-3-((3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)thioureid-
o)methyl)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00456##
[0203] To
N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-1H-imidazole-1-carbot-
hioamide (0.54 g, 2 mmol) in N,N-dimethylformamide (20 mL) was
added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (0.35 g, 2 mmol). The reaction was stirred at 50.degree. C. for
3 hours. The reaction was cooled and concentrated to yield crude
product. The crude material was purified via flash chromatography
(50-100% ethyl acetate-hexane) yielding the first spot/fractions
(TLC) as the product. The fractions were combined and concentrated
to yield
(3-hydroxy-3-((3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)thioureido)methy-
l)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (0.51 g, 1.39
mmol, 68.2% yield) as a white powder. .sup.1H NMR (500 MHz,
DMSO-D6) .delta. ppm 11.46 (s, 1H), 5.26 (s, 1 H), 3.89 (dd,
J=13.58, 5.34 Hz, 1H), 3.68 (dd, J=13.73, 4.88 Hz, 1H), 2.55-2.94
(m, 8H), 2.06 (dd, J=9.31, 3.20 Hz, 1H), 1.66-1.90 (m, 6H),
1.21-1.59 (m, 4H); [M+H].sup.+=365.1.
Step C:
(2-(4,5,6,7-Tetrahydrobenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospir-
o[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00457##
[0205] To
(3-hydroxy-3-((3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)thioure-
ido)methyl)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (0.5
g, 1.37 mmol) in N,N-dimethylformamide (20 mL) was added
N,N-diisopropylcarbodiimide (0.74 mL, 4.78 mmol). The reaction was
stirred at 70.degree. C. for 4 hours. The reaction was concentrated
to yield a crude residue. The crude material was purified via flash
chromatography (50-100% ethyl acetate-hexanes) yielding the second
spot/fractions (TLC) as the product. The fractions were combined
and concentrated to yield
(2-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazo-
le-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.38 g, 1.14
mmol, 84% yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-D6)
.delta. ppm 8.26-8.86 (m, 1H), 3.78 (d, J=9.46 Hz, 1H), 3.66 (d,
J=9.77 Hz, 1H), 3.19-3.30 (m, J=14.95, 2.14 Hz, 1H), 2.96-3.12 (m,
2H), 2.78-2.94 (m, 3H), 2.54-2.65 (m, 4H), 2.19 (s, 1H), 2.00 (s,
1H), 1.66-1.85 (m, 7H), 1.43 (m, 3H). MS (LC/MS) R.T.=2.50;
[M+H-BH.sub.3].sup.+=319.1.
Step D:
N-(4,5,6,7-Tetrahydrobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole--
5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00458##
[0207] To
(2-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-ylamino)-4H-1'-ammoniosp-
iro[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.36
g, 1.08 mmol) in acetone (9 mL) was added 3M HCl (0.36 mL, 1.08
mmol). The reaction was stirred at room temperature for 4 hours.
The reaction was complete by TLC (lower spot). Ethyl acetate was
added and the aqueous layer was then separated. The aqueous layer
was neutralized with 1N sodium hydroxide. The product was extracted
with ethyl acetate (2.times.40 mL). The organics were combined,
dried with magnesium sulfate, filtered, and concentrated in vacuo
to afford racemic
N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine (0.28 g, 0.84 mmol, 78% yield) as a
white powder. .sup.1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.30-8.75
(br.s, 1H), 3.79 (d, J=9.44 Hz, 1H), 3.55 (d, J=9.76 Hz, 1H),
2.90-3.03 (m, 2H), 2.54-2.84 (m, 8H), 2.00 (s, 1H), 1.81-1.93 (m, 1
H), 1.75 (d, J=2.44 Hz, 4H), 1.36-1.67 (m, 3H). MS (LC/MS)
R.T.=1.39; [M+H].sup.+=319.1.
[0208] The enantiomers were separated using a Chiralpak AD-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
23% methanol (0.1% DEA) in CO.sub.2. The wavelength was set at 300
nM. The separated peaks were concentrated in vacuo to yield white
powders. The first peak off the column was
(S)--N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine (0.1 g, 0.29 mmol, 36.8% yield).
(14a, S-isomer): .sup.1H NMR (500 MHz, DMSO-D6) .delta. ppm
8.29-8.89 (br.s, 1H), 3.80 (d, J=9.46 Hz, 1H), 3.54 (d, J=9.77 Hz,
1H), 2.94-3.04 (m, 2H), 2.55-2.85 (m, 8H), 2.00 (s, 1H), 1.85-1.94
(m, 1 H), 1.70-1.81 (m, J=2.43 Hz, 4H), 1.39-1.65 (m, 3H). MS
(LC/MS) R.T.=1.54; [M+H].sup.+=319.1. The second peak was
(R)--N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine (0.11 g, 0.30 mmol, 38.2% yield).
(14b, R-isomer): .sup.1H NMR (500 MHz, DMSO-D6) .delta. ppm
8.24-8.96 (br.s, 1H), 3.79 (d, J=9.46 Hz, 1H), 3.52 (d, J=9.76 Hz,
1H), 2.95-3.08 (m, 2H), 2.55-2.80 (m, 8H), 2.00 (s, 1H), 1.85-1.90
(m, 1H), 1.70-1.79 (m, J=2.42 Hz, 4 H), 1.39-1.63 (m, 3H);
[M+H].sup.+=319.1.
Example 15
N-(4-Isopropylthiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octa-
n]-2-amine
##STR00459##
[0209] Step A:
N-(4-Isopropylthiazol-2-yl)-1H-imidazole-1-carbothioamide
##STR00460##
[0211] To 4-isopropylthiazol-2-amine (1.04 g, 7.31 mmol) in
acetonitrile (30 mL) was added 1,1-thiocarbonyldiimidazole (1.7 g,
9.5 mmol). The reaction was stirred at 50.degree. C. for 18 hours.
The reaction was cooled to room temperature and the precipitate was
filtered and washed with acetonitrile (2.times.50 mL). The yellow
powder was dried in a vacuum oven (40.degree. C.) for 2 hours. The
product, N-(4-isopropylthiazol-2-yl)-1H-imidazole-1-carbothioamide
(1.02 g, 4.04 mmol, 55.3% yield), was taken directly to the next
step without any further purification.
Step B:
(3-Hydroxy-3-((3-(4-isopropylthiazol-2-yl)thioureido)methyl)-1-amm-
oniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00461##
[0213] To N-(4-isopropylthiazol-2-yl)-1H-imidazole-1-carbothioamide
(0.57 g, 2.26 mmol) in N,N-dimethylformamide (20 mL) was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (0.38 g, 2.26 mmol). The reaction was stirred at 70.degree. C.
for 24 hours. The reaction was cooled and concentrated to yield
crude product. The crude material was purified via flash
chromatography (50-100% ethyl acetate-hexane) to yield
(3-hydroxy-3-((3-(4-isopropylthiazol-2-yl)thioureido)methyl)-1-ammoniobic-
yclo[2.2.2]octan-1-yl)trihydroborate (0.56 g, 1.58 mmol, 70.0%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 11.62 (s, 1H), 6.70 (s, 1H), 5.31 (s, 1H), 3.58-3.96
(m, 2H), 2.63-3.11 (m, 7H), 2.00-2.22 (m, 1H), 1.65-1.97 (m, 3H),
1.16-1.61 (m, 10H). MS (LC/MS) R.T.=3.43; [M+H].sup.+=353.2.
Step C:
(2-(5-Isopropylthiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'--
bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00462##
[0215] To
(3-hydroxy-3-((3-(5-isopropylthiazol-2-yl)thioureido)methyl)-1-a-
mmoniobicyclo[2.2.2]octan-1-yl)trihydroborate (0.54 g, 1.52 mmol)
in N,N-dimethylformamide (20 mL) was added
N,N'-diisopropylcarbodiimide (0.83 mL, 5.33 mmol). The reaction was
stirred at 50.degree. C. for 24 hours. The reaction was
concentrated to yield a crude residue. The crude material was
purified via flash chromatography (50-100% ethyl acetate-hexanes)
yielding the second spot/fractions (TLC) as the product. The
fractions were combined and concentrated to yield
(2-(5-isopropylthiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bicyclo-
[2.2.2]octane]-1'-yl)trihydroborate (0.39 g, 1.22 mmol, 80% yield)
as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
6.55 (s, 1H), 3.75 (m, 2H), 3.20-3.31 (m, 1H), 2.96-3.15 (m, 2H),
2.77-2.97 (m, 4H), 2.22 (s, 1H), 2.01 (s, 1H), 1.68-1.90 (m, 3H),
1.43 (s, 3H), 1.21 (d, J=7.02 Hz, 6H). MS (LC/MS) R.T.=2.36;
[M+H-BH.sub.3].sup.+=307.2.
Step D:
N-(4-Isopropylthiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.-
2.2]octan]-2-amine
##STR00463##
[0217] To
(2-(4-isopropylthiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3-
'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.42 g, 1.31 mmol) in
acetone (9 mL) was added 3M HCl (0.44 mL, 1.31 mmol). The reaction
was stirred at room temperature for 4 hours. The reaction was
complete by TLC (lower spot). Ethyl acetate was added and the
aqueous layer was then separated. The aqueous layer was neutralized
with 1N sodium hydroxide. The product was extracted with ethyl
acetate (2.times.40 mL). The organics were combined, dried with
magnesium sulfate, filtered, and concentrated in vacuo to afford
racemic
N-(4-isopropylthiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oct-
an]-2-amine (0.34 g, 1.05 mmol, 80% yield) as a white powder.
.sup.1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.20-8.91 (m, 1H), 6.52
(s, 1H), 3.83 (d, J=9.46 Hz, 1H), 3.57 (d, J=9.46 Hz, 1H), 2.99 (s,
2H), 2.58-2.92 (m, 5H), 2.02 (s, 1H), 1.82-1.96 (m, 1H), 1.38-1.66
(m, 3H), 1.20 (d, J=7.02 Hz, 6H). MS (LC/MS) R.T.=1.27;
[M+H].sup.+=307.1.
[0218] The enantiomers were separated using a Chiralcel OJ-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
23% methanol (0.1% DEA) in CO.sub.2. The wavelength was set at 300
nM. The separated peaks were concentrated in vacuo to yield white
powders. The first peak off the column was
(S)--N-(4-isopropylthiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (0.01 g, 0.03 mmol, 2.95% yield). (15a, S-isomer):
.sup.1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.35-8.95 (m, 1H), 6.52
(s, 1H), 3.82 (d, J=9.41 Hz, 1H), 3.57 (d, J=9.46 Hz, 1H), 2.99 (s,
2H), 2.60-2.88 (m, 5H), 2.02 (s, 2H), 1.82-1.96 (m, 1H), 1.38-1.65
(m, 3H), 1.20 (d, J=6.71 Hz, 6H). MS (LC/MS) R.T.=1.38;
[M+H].sup.+=307.1. The second peak was
(R)--N-(4-isopropylthiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (0.02 g, 0.05 mmol, 4.83% yield). (15b, R-isomer):
.sup.1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.35-8.91 (m, 1H), 6.52
(s, 1H), 3.82 (d, J=9.44 Hz, 1H), 3.57 (d, J=9.46 Hz, 1H), 2.99 (s,
2H), 2.60-2.90 (m, 5H), 2.02 (s, 2H), 1.82-1.96 (m, 1H), 1.38-1.62
(m, 3H), 1.20 (d, J=6.78 Hz, 6H). MS (LC/MS) R.T.=1.49;
[M+H].sup.+=307.3.
Example 16
N-(Thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00464##
[0219] Step A: N-(Thiazol-2-yl)-1H-imidazole-1-carbothioamide
##STR00465##
[0221] To thiazol-2-amine (2.12 g, 21.17 mmol) in acetonitrile (30
mL) and tetrahydrofuran (5 mL) was added
di(1H-imidazol-1-yl)methanethione (4.90 g, 27.5 mmol). The reaction
was stirred at 60.degree. C. for 5 hours. The reaction was cooled
to room temperature and the precipitate was filtered and washed
with cold acetonitrile (2.times.15 mL) to afford an orange-brown
powder. The product, N-(thiazol-2-yl)-1H-imidazole-1-carbothioamide
(3.70 g, 17.60 mmol, 83% yield), was taken directly to the next
step without any further characterization.
Step B:
(3-Hydroxy-3-((3-thiazol-2-ylthioureido)methyl)-1-ammoniobicyclo[2-
.2.2]octan-1-yl)trihydroborate
##STR00466##
[0223] To N-(thiazol-2-yl)-1H-imidazole-1-carbothioamide (1.7 g, 8
mmol) in N,N-dimethylformamide (30 mL) was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (1.37 g, 8 mmol). The reaction was stirred at 50.degree. C. for
4 hours. The reaction was cooled and concentrated to yield crude
product. The crude material was purified via flash chromatography
(50-100% ethyl acetate-hexane) yielding the first spot/fractions
(TLC) as the product. The fractions were combined and concentrated
to yield
(3-hydroxy-3-((3-thiazol-2-ylthioureido)methyl)-1-ammoniobicyclo[2.2.2]oc-
tan-1-yl)trihydroborate (1.5 g, 4.80 mmol, 59.8% yield) as a white
powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 11.64 (s,
1H), 7.42 (d, J=3.36 Hz, 1H), 7.14 (m, 1H), 5.32 (s, 1H), 3.78 (dd,
2H), 2.59-3.02 (m, 6H), 1.99-2.18 (m, 1H), 1.79-1.92 (m, 2H),
1.64-1.80 (m, 1H), 1.19-1.65 (m, 4H). MS (LC/MS) R.T.=2.73;
[M+H].sup.+=311.1.
Step C:
(2-(Thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bicyclo[2.2-
.2]octane]-1'-yl)trihydroborate
##STR00467##
[0225] To
(3-hydroxy-3-((3-thiazol-2-ylthioureido)methyl)-1-ammoniobicyclo-
[2.2.2]octan-1-yl)trihydroborate (1.2 g, 3.84 mmol) in
N,N-dimethylformamide (20 mL) was added
N,N'-diisopropylcarbodiimide (2.09 mL, 13.45 mmol). The reaction
was stirred at 50.degree. C. for 24 hours. The reaction was
concentrated to yield a crude residue. The crude material was
purified via flash chromatography (50-100% ethyl acetate-hexane)
yielding the first spot/fractions (TLC) as the product. The
fractions were combined and concentrated to yield
(2-(thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bicyclo[2.2.2]octa-
ne]-1'-yl)trihydroborate (0.84 g, 3.02 mmol, 79% yield) as a white
powder. .sup.1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.37-9.14 (m,
1H), 7.32 (d, J=3.66 Hz, 1H), 7.04 (d, J=3.66 Hz, 2H), 3.79 (d,
J=10.07 Hz, 1H), 3.67 (d, J=10.07 Hz, 1H), 3.20-3.29 (m, J=14.95,
2.14 Hz, 1H), 2.97-3.15 (m, 2H), 2.78-2.94 (m, 3H), 2.22 (s, 1H),
1.95-2.08 (m, 1H), 1.66-1.85 (m, 3H), 1.43 (s, 3H). MS (LC/MS)
R.T.=1.57; [M+H-BH.sub.3].sup.+=265.1.
Step D:
N-(Thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]--
2-amine
##STR00468##
[0227] To
(2-(thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-bicyclo[2-
.2.2]octane]-1'-yl)trihydroborate (0.57 g, 2.05 mmol) in acetone (9
mL) was added 3M HCl (0.68 mL, 2.05 mmol). The reaction was stirred
at room temperature for 4 hours. The reaction was complete by TLC
(lower spot). Ethyl acetate was added and the aqueous layer was
then separated. The aqueous layer was neutralized with 1N sodium
hydroxide. The product was extracted with ethyl acetate (2.times.40
mL). The organics were combined, dried with magnesium sulfate,
filtered, and concentrated in vacuo to afford racemic
N-(thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine
(0.4 g, 1.44 mmol, 70.2% yield) as a white powder. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 8.64 (s, 1H), 7.24 (d, J=3.74 Hz,
2H), 7.01 (d, J=3.75 Hz, 2H), 3.78 (d, J=9.80 Hz, 2H), 3.53 (d,
J=9.80 Hz, 2H), 2.97-3.05 (m, 4H), 2.74-2.86 (m, 4H), 2.65 (t,
J=7.84 Hz, 4H), 2.01 (s, 2H), 1.88 (s, 2H), 1.53-1.64 (m, 4H),
1.45-1.56 (m, 2H). MS (LC/MS) R.T.=0.28; [M+H].sup.+=265.1.
[0228] The enantiomers were separated using a Chiralcel OJ-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
23% methanol (0.1% DEA) in CO.sub.2. The wavelength was set at 300
nM. The separated peaks were concentrated in vacuo to yield white
powders. The first peak off the column was
(S)--N-(thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2--
amine (0.12 g, 0.43 mmol, 18.80% yield). (16a, S-isomer): .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 8.64 (s, 1H), 7.26 (d,
J=3.75 Hz, 2H), 7.00 (d, J=3.77 Hz, 2H), 3.78 (d, J=9.81 Hz, 2H),
3.54 (d, J=9.82 Hz, 2H), 2.95-3.10 (m, 4H), 2.75-2.82 (m, 4H), 2.65
(t, J=7.80 Hz, 4H), 2.01 (s, 2H), 1.88 (s, 2H), 1.53-1.60 (m, 4H),
1.42-1.51 (m, 2H). MS (LC/MS) R.T.=0.32; [M+H].sup.+=265.1. The
second peak was
(R)--N-(thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2--
amine (0.15 g, 0.52 mmol, 22.96% yield). (16b, R-isomer): .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 8.64 (s, 1H), 7.25 (d,
J=3.74 Hz, 2H), 7.00 (d, J=3.76 Hz, 2H), 3.78 (d, J=9.80 Hz, 2H),
3.53 (d, J=9.80 Hz, 2H), 2.95-3.08 (m, 4H), 2.75-2.84 (m, 4H), 2.65
(t, J=7.80 Hz, 4H), 2.01 (s, 2H), 1.88 (s, 2H), 1.54-1.62 (m, 4H),
1.43-1.53 (m, 2H). MS (LC/MS) R.T.=0.28; [M+H].sup.+=265.3.
Example 17
N-(4-(4-Methoxyphenyl)-5-methylthiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine
##STR00469##
[0229] Step A:
N-(4-(4-Methoxyphenyl)-5-methylthiazol-2-yl)-1H-imidazole-1-carbothioamid-
e
##STR00470##
[0231] To 4-(4-methoxyphenyl)-5-methylthiazol-2-amine (0.98 g, 4.45
mmol) in acetonitrile (25 mL) was added
di(1H-imidazol-1-yl)methanethione (1.03 g, 5.78 mmol). The reaction
was stirred at 50.degree. C. for 3 hours. The reaction was cooled
to room temperature and the precipate was filtered. The powder was
washed with acetonitrile (2.times.10 mL) and dried to yield racemic
N-(4-(4-methoxyphenyl)-5-methylthiazol-2-yl)-1H-imidazole-1-carbothioamid-
e (1.28 g, 3.87 mmol, 87% yield) as a yellow powder. The product
was taken directly to the next step.
Step B:
(2-(4-(4-Methoxyphenyl)-5-methylthiazol-2-ylamino)-4H-1'-ammoniosp-
iro[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00471##
[0233] To
N-(4-(4-methoxyphenyl)-5-methylthiazol-2-yl)-1H-imidazole-1-carb-
oxamide (0.44 g, 1.39 mmol) in N,N-dimethylformamide (25 mL) was
added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (0.24 g, 1.39 mmol). The reaction was stirred at 70.degree. C.
for 2 hours. N,N'-Diisopropylcarbodiimide (0.65 mL, 4.16 mmol) was
added and the reaction heated to 75.degree. C. for 2 hours. The
reaction was cooled and concentrated to afford the crude product.
The crude material was purified via column chromatography (60-100%
ethyl acetate/hexanes) to yield
(2-(4-(4-methoxyphenyl)-5-methylthiazol-2-ylamino)-4H-1'-ammoniospi-
ro[oxazole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.41 g,
1.029 mmol, 74.2% yield) as a yellow powder. .sup.1H NMR (500 MHz,
DMSO-D6) .delta. ppm 7.59 (d, J=7.63 Hz, 2H), 6.98 (d, J=8.55 Hz,
2H), 3.80 (s, 4H), 3.67 (s, 1H), 3.24-3.31 (m, 1H), 3.13 (s, 1H),
3.03 (s, 1H), 2.83-2.92 (m, 3H), 2.39 (s, 3H), 2.23 (s, 1H), 2.07
(s, 1H), 1.73-1.82 (m, 2H), 1.44 (s, 1H). MS (LC/MS) R.T.=2.88;
[M+H].sup.+=399.34.
Step C:
N-(4-(4-Methoxyphenyl)-5-methylthiazol-2-yl)-4H-1'-azaspiro[oxazol-
e-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00472##
[0235] To
(3-hydroxy-3-((3-(4-(4-methoxyphenyl)-5-methylthiazol-2-yl)thiou-
reido)methyl)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
(0.08 g, 0.19 mmol) in acetone (9 mL) was added 2M HCl (0.09 mL,
0.19 mmol). The reaction was stirred at room temperature for 1
hour. The reaction was complete by TLC (lower spot). Ethyl acetate
was added and the aqueous layer was collected and neutralized with
1N sodium hydroxide. The product was extracted with ethyl acetate
(2.times.40 mL). The organics were combined, dried with magnesium
sulfate, filtered, and concentrated in vacuo to afford racemic
N-(4-(4-methoxyphenyl)-5-methylthiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine (0.05 g, 0.12 mmol, 66.8% yield) as a
white powder. .sup.1H NMR (500 MHz, DMSO-D6) .delta. ppm 7.58 (d,
J=8.24 Hz, 2H), 6.98 (d, J=8.55 Hz, 2H), 3.83 (d, J=9.16 Hz, 1H),
3.80 (s, 3H), 3.56 (d, J=9.46 Hz, 1H), 3.00 (s, 2H), 2.74-2.83 (m,
2H), 2.66 (t, J=7.63 Hz, 2H), 2.38 (s, 3H), 2.03 (s, 1H), 1.91 (s,
1H), 1.54-1.62 (m, 2H), 1.48 (d, J=7.02 Hz, 1H). MS (LC/MS)
R.T.=2.04; [M+H].sup.+=385.28.
Example 18
(E)-N-(1'-Azaspiro[oxazolidine-5,3'-bicyclo[2.2.2]octane]-2-ylidene)pyridi-
n-3-amine
##STR00473##
[0236] Step A:
(3-Hydroxy-3-((3-pyridin-3-ylthioureido)methyl)-1-ammoniobicyclo[2.2.2]oc-
tan-1-yl)trihydroborate
##STR00474##
[0238] To a stirring suspension of
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (247 mg, 1.45 mmol) in tetrahydrofuran (3 mL) was added a
solution of 3-isothiocyanatopyridine (298 mg, 2.19 mmol) in
tetrahydrofuran (1.5 mL) and the reaction mixture was stirred at
room temperature overnight. The reaction mixture was evaporated in
vacuo and the residue (white waxy solid) purified via column
chromatography (3% methanol/ethyl acetate) to yield
(3-hydroxy-3-((3-pyridin-3-ylthioureido)methyl)-1-ammoniobicyclo[2.-
2.2]octan-1-yl)trihydroborate (294.6 mg, 0.96 mmol, 66.2% yield) as
a white foam. .sup.1H NMR (500 MHz, MeOD-d.sub.4) .delta. ppm
8.56-8.70 (m, 1H), 8.24-8.36 (m, 1H), 8.08-8.16 (m, 1H), 7.32-7.48
(m, 1H), 4.02-4.18 (m, 1H), 3.67-3.77 (m, 1H), 3.53-3.62 (m, 1H),
2.88-3.11 (m, 3H), 2.70-2.88 (m, 1H), 2.15-2.29 (m, 1H), 1.92-2.12
(m, 2H), 1.73-1.89 (m, 1H), 1.54-1.69 (m, 2H). MS (LC/MS)
R.T.=1.00; [MH.sup.+-BH.sub.3]=293.10.
Step B:
(E)-(2-(Pyridin-3-ylimino)-1'-ammoniospiro[oxazolidine-5,3'-bicycl-
o[2.2.2]octane]-1'-yl)trihydroborate
##STR00475##
[0240] To a solution of
(3-hydroxy-3-((3-pyridin-3-ylthioureido)methyl)-1-ammoniobicyclo[2.2.2]oc-
tan-1-yl)trihydroborate (294.6 mg, 0.96 mmol) in
N,N-dimethylformamide (5 mL) was added a solution of
N,N'-methanediylidenedipropan-2-amine (121 mg, 0.96 mmol) in
N,N-dimethylformamide (1 mL) and the reaction mixture was allowed
to stand at room temperature for 7 days. An additional 133 mg
N,N'-methanediylidenedipropan-2-amine in 0.5 mL
N,N-dimethylformamide was added and the reaction was left to
continue for another 7 days. The reaction was purified via column
chromatography (5-10% methanol/ethyl acetate) to afford
(E)-(2-(pyridin-3-ylimino)-1'-ammoniospiro[oxazolidine-5,3'-bicyclo[2.2.2-
]octane]-1'-yl)trihydroborate (134.3 mg, 0.49 mmol, 51.3% yield).
.sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.34-8.60 (m, 1H), 8.16 (d,
J=4.27 Hz, 1H), 7.59-7.96 (m, 1H), 7.35 (dd, J=8.24, 4.88 Hz, 1H),
3.89 (br. s., 1H), 3.65 (d, J=9.46 Hz, 1H), 3.16-3.23 (m, 1H),
3.04-3.16 (m, 1H), 2.85-3.04 (m, 2H), 2.23 (br. s., 1H), 1.74-1.97
(m, 4H), 1.36-1.70 (m, 2H). MS (LC/MS) R.T.=0.65;
[M+H-BH.sub.3].sup.+=259.21.
Step C:
(E)-N-(1'-Azaspiro[oxazolidine-5,3'-bicyclo[2.2.2]octane]-2-yliden-
e)pyridin-3-amine
##STR00476##
[0242] To a suspension of
(E)-(2-(pyridin-3-ylimino)-1'-ammoniospiro[oxazolidine-5,3'-bicyclo[2.2.2-
]octane]-1'-yl)trihydroborate (127 mg, 0.47 mmol) in acetone (5 mL)
was added 3M hydrochloric acid (2 mL, 6.00 mmol) and the mixture
was allowed to stand at room temperature for 2 hrs. It was then
added it to a separatory funnel containing water and chloroform.
The layers were separated, then the aqueous layer was made basic
with sodium carbonate solution and the mixture was re-extracted
with chloroform. Finally, the aqueous phase was washed with ethyl
acetate. The organics were combined, dried with magnesium sulfate,
filtered, and concentrated in vacuo. The residue was purified by
column chromatography (1% ammonium hydroxide/9% methanol/90%
dichloromethane) to afford racemic
(E)-N-(1'-azaspiro[oxazolidine-5,3'-bicyclo[2.2.2]octane]-2-ylidene)pyrid-
in-3-amine (19 mg, 0.074 mmol, 15.8% yield) as a white solid.
.sup.1H NMR (500 MHz, MeOD-d.sub.4) .delta. ppm 8.39 (br. s., 1H),
8.14 (d, J=4.27 Hz, 1H), 7.72 (br. s., 1H), 7.34 (dd, J=8.24, 4.88
Hz, 1H), 3.89 (d, J=9.77 Hz, 1H), 3.57 (d, J=10.38 Hz, 1H),
3.14-3.27 (m, 1H), 3.00-3.13 (m, 1H), 2.70-3.00 (m, 4H), 1.97-2.22
(m, 2H), 1.54-1.84 (m, 3 H). MS (LC/MS) R.T.=0.26;
[M+H].sup.+=259.16.
Example 19
N-(Pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00477##
[0243] Step A: 2-Bromo-6-isothiocyanatopyridine
##STR00478##
[0245] A mixture of 6-bromopyridin-2-amine (253 mg, 1.46 mmol),
chloroform (2 mL), sodium bicarbonate (850 mg, 10.12 mmol) and
water (3 mL) was assembled, and to this was added a solution of
thiophosgene (190 mg, 1.65 mmol) in chloroform (1 mL). The reaction
mixture was stirred at room temperature for 4 hours. The reaction
mixture was transferred to a separatory funnel and partitioned
between ethyl acetate and water. The organic layer was washed with
brine, dried over magnesium sulfate, filtered and the solvent
evaporated to give a yellow solid. The solid was purified by column
chromatography (5% ethyl acetate/hexanes) to afford
2-bromo-6-isothiocyanatopyridine (281 mg, 1.31 mmol, 89% yield) as
a white solid. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm
7.50-7.62 (m, 1H), 7.34-7.43 (m, 1H), 6.91-7.11 (m, 1 H). MS
(LC/MS) R.T.=1.92; [M+H].sup.+=216.86.
Step B:
N-(6-Bromopyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]-
octan]-2-amine
##STR00479##
[0247] To a solution of 2-bromo-6-isothiocyanatopyridine (281 mg,
1.31 mmol) in N,N-dimethylformamide (8 mL) and Hunig's base (0.6
mL, 3.44 mmol) was added (+/-) 3-(aminomethyl)quinuclidin-3-ol
dihydrochloride (300 mg, 1.31 mmol) and the resulting mixture was
heated to 75.degree. C. for 2.5 hrs. MS (LC/MS) R.T.=1.04;
[M+H].sup.+=373.01. To this reaction mixture was added
di-isopropyl-carbodiimide (523 mg, 4.14 mmol) and the heating at
75.degree. C. was continued for 2.25 hours. The mixture was allowed
to cool to room temperature over the weekend. The reaction mixture
was concentrated in vacuo. The material was purified by column
chromatography then preparative HPLC to afford
N-(6-bromopyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]--
2-amine (291.5 mg, 0.86 mmol, 66.2 5 yield) as a yellow solid
(containing 2-amino-6-bromopyridine impurity). MS (LC/MS)
R.T.=0.65; [M+H].sup.+=337.0.
Step C:
N-(Pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]--
2-amine
##STR00480##
[0249] To
N-(6-bromopyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (291 mg, 0.86 mmol) in methanol (20 mL) was
hydrogenated over 10% palladium on carbon (23 mg) in the Parr
apparatus for 2 hours. The catalyst was removed by filtration and
the filtrated concentrated in vacuo. The residue was purified by
column chromatography (0.7% ammonium hydroxide/6.3% methanol/93%
chloroform) to give racemic
N-(pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine
(41.3 mg, 0.16 mmol, 18.6 5 yield). .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.13-8.33 (m, 1H), 7.57-7.72 (m, 1H), 6.83-7.04 (m,
2H), 3.97 (d, J=10.07 Hz, 1H), 3.66 (d, J=10.07 Hz, 1H), 3.18-3.27
(m, 1H), 3.03-3.14 (m, 1H), 2.94 (t, J=7.63 Hz, 2H), 2.70-2.90 (m,
2H), 2.06-2.24 (m, 2H), 1.57-1.87 (m, 3H). MS (LC/MS) R.T.=1.76;
[M+H].sup.+=259.25.
Example 20
N-(4-(4-Methoxyphenyl)thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.-
2.2]octan]-2-amine
##STR00481##
[0250] Step A:
N-(5-(4-Methoxyphenyl)thiazol-2-yl)-1H-imidazole-1-carbothioamide
##STR00482##
[0252] To 5-(4-methoxyphenyl)thiazol-2-amine (1.07 g, 5.19 mmol) in
acetonitrile (30 mL) and tetrahydrofuran (5 mL) was added
di(1H-imidazol-1-yl)methanethione (1.20 g, 6.74 mmol). The reaction
was stirred at 60.degree. C. for 18 hours. The reaction was cooled
to room temperature and the precipitate was filtered. The powder
was washed with cold acetonitrile (2.times.15 mL) and dried to
yield
N-(5-(4-methoxyphenyl)thiazol-2-yl)-1H-imidazole-1-carbothioamide
(0.59 g, 1.86 mmol, 35.9% yield) as a orange-brown powder. The
product was taken directly to the next step without any further
characterization.
Step B:
(3-Hydroxy-3-((3-(4-(4-methoxyphenyl)thiazol-2-yl)thioureido)methy-
l)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate
##STR00483##
[0254] To
N-(4-(4-methoxyphenyl)thiazol-2-yl)-1H-imidazole-1-carbothioamid- e
(0.57 g, 1.82 mmol) in N,N-dimethylformamide (20 mL) was added
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydrobora-
te (0.31 g, 1.82 mmol). The reaction was stirred at 60.degree. C.
for 4 hours. The reaction was cooled and concentrated to yield
crude product. The crude material was purified via flash
chromatography (60-100% ethyl acetate-hexane) yielding the first
spot/fractions (TLC) as the product. The fractions were combined
and concentrated in vacuo to yield
(3-hydroxy-3-((3-(4-(4-methoxyphenyl)thiazol-2-yl)thioureido)methyl)-1-am-
moniobicyclo[2.2.2]octan-1-yl)trihydroborate (0.6 g, 1.43 mmol, 79%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 11.74 (s, 1H), 7.91 (d, J=8.55 Hz, 2H), 7.43 (s, 1H),
6.97 (d, J=8.85 Hz, 2H), 5.50 (s, 1H), 3.62-3.98 (m, 5H), 2.70-3.10
(m, 6H), 2.13 (s, 1H), 1.94 (s, 1H), 1.66-1.91 (m, 2H), 1.11-1.62
(m, 4H). MS (LC/MS) R.T.=3.54; [M+H].sup.+=417.1.
Step C:
(2-(4-(4-Methoxyphenyl)thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazo-
le-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate
##STR00484##
[0256] To
(3-hydroxy-3-((3-(4-(4-methoxyphenyl)thiazol-2-yl)thioureido)met-
hyl)-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydroborate (0.59 g, 1.41
mmol) in N,N-dimethylformamide (20 mL) was added
N,N'-diisopropylcarbodiimide (0.66 mL, 4.23 mmol). The reaction was
stirred at 70.degree. C. for 24 hours. The solvent was removed in
vacuo and the residue was purified by flash chromatography (50-100%
ethyl acetate-hexanes), collecting the first component as the
product, to yield
(2-(4-(4-methoxyphenyl)thiazol-2-ylamino)-4H-1'-ammoniospiro[oxazole-5,3'-
-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.45 g, 1.17 mmol, 83%
yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 8.65 (s, 1H), 7.89 (d, J=7.32 Hz, 2 H), 7.15-7.43 (m,
1H), 6.95 (d, J=8.85 Hz, 2H), 3.64-3.93 (m, 5H), 3.23-3.31 (m,
J=1.53 Hz, 1H), 3.09-3.21 (m, 1H), 2.99-3.09 (m, 1H), 2.79-2.97 (m,
3H), 2.25 (s, 1H), 1.96-2.16 (m, 1H), 1.68-1.91 (m, 3H), 1.45 (s,
3H). MS (LC/MS) R.T.=2.80; [M+H-BH.sub.3].sup.+=371.1.
Step D:
N-(4-(4-Methoxyphenyl)thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine
##STR00485##
[0258] To
(2-(4-(4-methoxyphenyl)thiazol-2-ylamino)-4H-1'-ammoniospiro[oxa-
zole-5,3'-bicyclo[2.2.2]octane]-1'-yl)trihydroborate (0.41 g, 1.07
mmol) in acetone (9 mL) was added 3M HCl (0.36 mL, 1.07 mmol). The
reaction was stirred at room temperature for 4 hours. The reaction
was complete by TLC (lower spot). Ethyl acetate was added and the
aqueous layer was then separated. The aqueous layer was neutralized
with 1N sodium hydroxide. The product was extracted with ethyl
acetate (2.times.40 mL). The organics were combined, dried with
magnesium sulfate, filtered, and concentrated in vacuo to afford
racemic
N-(4-(4-methoxyphenyl)thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine (0.3 g, 0.77 mmol, 72.1% yield) as a white
powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.56 (s,
1H), 7.77-8.00 (m, J=8.55 Hz, 2H), 7.27 (s, 1H), 6.80-7.08 (m, 2H),
3.87 (d, J=9.77 Hz, 1 H), 3.79 (s, 3H), 3.61 (d, J=9.77 Hz, 1H),
3.02 (s, 3H), 2.60-2.92 (m, 4H), 2.06 (s, 2H), 1.82-2.00 (m, 1H),
1.39-1.70 (m, 3H). MS (LC/MS) R.T.=1.95; [M+H].sup.+=371.2.
[0259] The enantiomers were separated using a Chiralpak AD-H
(30.times.250 mm, 5 .mu.m) column with a mobile phase consisting of
30% methanol (0.1% DEA) in CO.sub.2. The wavelength was set at 220
nM. The separated peaks were concentrated in vacuo to yield white
powders. The first peak off the column was
(S)--N-(4-(4-methoxyphenyl)thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine (0.035 g, 0.09 mmol, 22.87% yield).
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.56 (1H, br. s.),
7.86 (2H, d, J=8.55 Hz), 7.25 (1H, s), 6.93-6.96 (2H, m), 3.86 (1
H, d, J=9.77 Hz), 3.78 (3H, s), 3.60 (1H, d, J=9.46 Hz), 3.01 (2H,
s), 2.72-2.84 (2H, m), 2.62-2.71 (2H, m), 2.05 (1H, br. s.), 1.91
(1H, br. s.), 1.55-1.64 (2H, m), 1.44-1.52 (1H, m). MS (LC/MS)
R.T.=2.03; [M+H].sup.+=371.3. The second peak was
(R)--N-(4-(4-methoxyphenyl)thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine (0.055 g, 0.15 mmol, 35.9% yield). (21b,
R-isomer): .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.58
(1H, br. s.), 7.87 (2H, d, J=8.55 Hz), 7.26 (1H, s), 6.92-6.97 (2H,
m), 3.86 (1H, d, J=9.77 Hz), 3.78 (3H, s), 3.60 (1H, d, J=9.77 Hz),
3.01 (2H, s), 2.73-2.85 (2H, m), 2.63-2.71 (2H, m), 2.05 (1H, br.
s.), 1.91 (1H, br. s.), 1.54-1.64 (2H, m), 1.43-1.53 (1H, m). MS
(LC/MS) R.T.=2.03; [M+H].sup.+=371.3.
Example 21
(R)--N-(2-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)ben-
zo[d]thiazol-6-yl)acetamide
##STR00486##
[0260] Step A:
(3-((Benzyloxycarbonylamino)methyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octa-
n-1-yl)trihydroborate
##STR00487##
[0262] To
(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)tri-
hydroborate (10 g, 47.0 mmol) in dichloromethane (150 mL) was added
sodium carbonate (200 mL, 200 mmol) and benzyl chloroformate (9.5
mL, 66.5 mmol). The reaction mixture was stirred at room
temperature for 40 minutes. Dichloromethane and water were added
and the aqueous layer was then separated and extracted again with
dichloromethane (2.times.). The organic layers were combined, dried
over sodium sulfate, filtered and concentrated. The residue was
purified via flash chromatography (12-100% ethyl acetate-hexanes).
The product fractions were combined and concentrated to yield
racemic
(3-((benzyloxycarbonylamino)methyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octa-
n-1-yl)trihydroborate (3 g, 9.86 mmol, 20.96% yield) as a clear
oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 7.30-7.43 (5H,
m), 5.27 (1H, br. s.), 5.12 (2H, s), 3.36 (2H, d, J=6.04 Hz),
2.76-3.21 (6H, m), 2.21 (1H, br. s.), 1.97 (1H, br. s.), 1.71-1.86
(2H, m).
[0263] The enantiomers were separated using a Chiralpak OJ-H
(5.times.25) column with a mobile phase consisting of 20%
acetonitrile/methanol (1:1) in CO.sub.2. The wavelength was set at
210 nM. The first peak off the column was
(R)-(3-(aminomethyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]octan-1-yl)trihydro-
borate (37.67 g, 123 mmol) as a colorless oil. Optical rotation:
+28.2, c=2.9 in chloroform. The second peak off the column was
(S)-(3-((benzyloxycarbonylamino)methyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]-
octan-1-yl)trihydroborate (46.82 g, 153 mmol) as a light amber oil.
Optical rotation: -27.4, c=2.5 in chloroform.
Step B: (S)-3-(Aminomethyl)quinuclidin-3-ol, 2HCl
##STR00488##
[0265] A solution of
(S)-(3-((benzyloxycarbonylamino)methyl)-3-hydroxy-1-ammoniobicyclo[2.2.2]-
octan-1-yl)trihydroborate (20.5 g, 67 mmol) in acetone (120 mL) was
cooled on an ice bath. 3M Aqueous HCl (120 mL, 360 mmol) was added
over 2 minutes. Vigorous bubbling was observed. After 10 minutes,
the cold ice bath was removed and the mixture was allowed to warm
to room temperature. After 20 minutes, it was diluted with methanol
(800 mL) and flushed with nitrogen. Palladium on carbon (2 g, 1.88
mmol) was added and the reaction was flushed with nitrogen and
fitted with balloon of hydrogen. The reaction mixture was stirred
at room temperature overnight. It was then flushed with nitrogen
and filtered through a pad of Celite using methanol. The solvent
was evaporated to yield a crude yellow solid. The solids were
dissolved in water (25 mL), then ethanol (400 mL) was added. White
crystals formed immediately. They were collected by filtration and
washed with ethanol, followed by ether. A white crystalline solid
was obtained, (S)-3-(aminomethyl)quinuclidin-3-ol, 2HCl (10.7 g,
46.7 mmol, 69.3% yield). .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 10.93 (1H, br. s.), 8.24 (3H, br. s.), 6.02 (1H, s),
3.26 (1H, d, J=13.43 Hz), 2.99-3.22 (5H, m), 2.10-2.19 (2H, m),
1.81-1.90 (1H, m), 1.72-1.81 (1H, m), 1.60-1.72 (1H, m). Optical
rotation: [.alpha.].sup.20.sub.D=-50.9.degree. (c=6.4, water).
Step C:
N-(2-(1H-Imidazole-1-carbothioamido)benzo[d]thiazol-6-yl)acetamide
##STR00489##
[0267] To N-(2-aminobenzo[d]thiazol-6-yl)acetamide (4 g, 19.3 mmol)
in acetonitrile (100 mL) was added
di(1H-imidazol-1-yl)methanethione (3.44 g, 19.30 mmol). The
reaction was allowed to stir at 80.degree. C. overnight. The
reaction was cooled to room temperature and the precipitate was
filtered. The product,
N-(2-(1H-imidazole-1-carbothioamido)benzo[d]thiazol-6-yl)acetamide
(3.6 g, 11.34 mmol, 58.8 5 yield), was taken directly to the next
step without any further purification.
Step D:
(R)--N-(2-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-yla-
mino)benzo[d]thiazol-6-yl)acetamide
##STR00490##
[0269] To
N-(2-(1H-imidazole-1-carbothioamido)benzo[d]thiazol-6-yl)acetami-
de (300 mg, 0.95 mmol) in N,N-dimethylformamide (10 mL) was added
(S)-3-(aminomethyl)quinuclidin-3-ol, 2HCl (238 mg, 1 mmol) and
triethylamine (0.39 mL, 2.84 mmol). The reaction was heated to
80.degree. C. for 3 hours. N,N'-diisopropylcarbodiimide (0.59 mL,
3.78 mmol) was then added to the reaction mixture. The mixture was
heated at 80.degree. C. for another 2 hours. The reaction was
cooled, then chloroform and water were added to the mixture. The
organic layer concentrated in vacuo to yield crude product. The
crude material was purified via flash chromatography (2-20% [10%
ammonium hydroxide/methanol-chloroform). The product fractions were
then triturated with ether to yield
(R)--N-(2-(4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)be-
nzo[d]thiazol-6-yl)acetamide (144.5 mg, 0.39 mmol, 41.2% yield) as
a white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm 9.98
(1H, s), 8.93 (1H, br. s.), 8.12 (1H, d, J=1.83 Hz), 7.52 (1H, d,
J=8.42 Hz), 7.38 (1H, dd, J=8.60, 2.01 Hz), 3.88 (1H, d, J=9.88
Hz), 3.62 (1H, d, J=9.88 Hz), 3.02 (2H, s), 2.74-2.85 (2H, m), 2.66
(2H, t, J=7.68 Hz), 2.05 (4H, s), 1.91 (1H, br. s.), 1.41-1.64 (3H,
m). MS (LC/MS) R.T.=1.55; [M+H].sup.+=372.2.
Example 22
(R)--N-(6-(Difluoromethoxy)benzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine
##STR00491##
[0270] Step A: 6-(Difluoromethoxy)benzo[d]thiazol-2-amine
##STR00492##
[0272] To 4-(difluoromethoxy)aniline (9.55 g, 60 mmol) in acetic
acid (90 mL) was added potassium thiocyanate (KSCN) (12.41 mL, 240
mmol). The mixture was stirred for 20 minutes (KSCN dissolved into
solution). To this mixture bromine (3.08 mL, 60.0 mmol) in acetic
acid (40 mL) was added dropwise over 20 minutes. The reaction was
stirred at room temperature overnight. It was poured into a mixture
of 800 ml ice water and 200 ml saturated ammonium hydroxide. The
product was extracted with ethyl acetate (5.times.). The organics
were combined, washed with brine, dried over sodium sulfate,
filtered and concentrated in vacuo to afford
6-(difluoromethoxy)benzo[d]thiazol-2-amine (12.6 g, 52.4 mmol, 87%
yield) as a yellow solid.
Step B:
N-(6-(Difluoromethoxy)benzo[d]thiazol-2-yl)-1H-imidazole-1-carboth-
ioamide
##STR00493##
[0274] To 6-(difluoromethoxy)benzo[d]thiazol-2-amine (0.5 g, 2.3
mmol) in acetonitrile (15 mL) was added
1,1'-thiocarbonyldiimidazole (0.49 g, 2.8 mmol). The reaction was
stirred at 70.degree. C. overnight. The reaction was cooled to room
temperature and the precipitate was filtered to yield
N-(6-(difluoromethoxy)benzo[d]thiazol-2-yl)-1H-imidazole-1-carbothioamide
(500 mg, 1.53 mmol, 66.3% yield) as a yellow solid.
Step C:
(R)--N-(6-(Difluoromethoxy)benzo[d]thiazol-2-yl)-4H-1'-azaspiro[ox-
azole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00494##
[0276] To a solution of
N-(6-(difluoromethoxy)benzo[d]thiazol-2-yl)-1H-imidazole-1-carbothioamide
(285 mg, 0.87 mmol) in N,N-dimethylformamide (5 mL) was added
(S)-3-(aminomethyl)quinuclidin-3-ol, 2HCl (200 mg, 0.87 mmol) and
triethylamine (0.4 mL, 2.87 mmol). The reaction was heated to
70.degree. C. for 2 hours. N,N'-diisopropylcarbodiimide (0.4 mL,
2.57 mmol) was then added to the reaction mixture. The mixture was
heated at 70.degree. C. for another 3 hours. It was cooled and then
poured into toluene/0.3M sodium hydroxide. The product was
extracted with toluene (4.times.) and chloroform (3.times.). The
organics were combined, washed with water (3.times.), dried over
sodium sulfate, filtered and concentrated in vacuo to yield crude
product. The crude material was purified via flash chromatography
(2-20% [10% ammonium hydroxide/methanol]-chloroform) to afford
(R)--N-(6-(difluoromethoxy)benzo[d]thiazol-2-yl)-4H-1'-azaspiro[ox-
azole-5,3'-bicyclo[2.2.2]octan]-2-amine (186.2 mg, 0.49 mmol, 55.5%
yield) as a white powder. M.P. 223-5.degree. C. .sup.1H NMR (500
MHz, DMSO-d.sub.6) .delta. ppm 8.99 (1H, br. s.), 7.69 (1H, d,
J=2.75 Hz), 7.61 (1H, d, J=8.55 Hz), 7.02-7.34 (2H, m), 3.89 (1H,
d, J=10.07 Hz), 3.64 (1H, d, J=9.77 Hz), 3.03 (2H, d, J=2.44 Hz),
2.73-2.86 (2H, m), 2.62-2.70 (2H, m), 2.07 (1H, br. s.), 1.92 (1H,
br. s.), 1.54-1.65 (2H, m), 1.44-1.53 (1 H, m). MS (LC/MS)
R.T.=1.43; [M+H].sup.+=381.1.
Example 23
(R)--N-(6-Methoxypyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00495##
[0277] Step A: 4-Isothiocyanato-6-methoxypyrimidine
##STR00496##
[0279] To a bright orange solution of
1,1'-thiocarbonyldipyridin-2(1H)-one (1.86 g, 7.99 mmol) in
dichloromethane at room temperature was added
6-methoxypyrimidin-4-amine (1 g, 8 mmol). The orange solution was
stirred at room temperature for 18 hours. The LC/MS showed the
desired product as one of the major peaks. The deep orange solution
was concentrated and the remaining residue was filtered. The
filtrate was purified by silica gel chromatography (10-50% ethyl
acetate/hexanes) to afford 4-isothiocyanato-6-methoxypyrimidine
(0.72 g, 4.3 mmol, 54% yield) as a yellow oil. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 8.49 (1H, d, J=5.79 Hz), 6.95 (1H,
d, J=5.79 Hz), 3.92 (3H, s). MS (LC/MS) R.T.=3.15;
[M+H].sup.+=168.1.
Step B:
(R)--N-(6-Methoxypyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00497##
[0281] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (from
Step B of Example 21) (0.34 g, 1.49 mmol) in N,N-dimethylformamide
(15 mL) was added Cs.sub.2CO.sub.3 (1.22 g, 3.74 mmol) and
4-isothiocyanato-6-methoxypyrimidine (0.25 g, 1.5 mmol). The
suspension was stirred at room temperature for 30 minutes.
N,N'-diisopropylcarbodiimide (0.7 mL, 4.5 mmol) was then added and
the mixture was stirred at room temperature for 18 hours. The
mixture was concentrated and purified by silica gel chromatography
(5-15% [9:1 methanol:ammonium hydroxide]/chloroform) to afford
(R)--N-(6-methoxypyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (0.21 g, 0.72 mmol, 48.2% yield) as a white solid.
M.P. 186-8.degree. C. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.40
(1H, s), 6.17 (1H, br. s.), 3.92-4.04 (1H, m), 3.89 (3H, s), 3.68
(1H, d, J=10.32 Hz), 3.12-3.23 (1H, m), 2.98-3.12 (1H, m),
2.67-2.97 (4H, m), 2.11 (2H, br. s.), 1.48-1.82 (3H, m). MS (LC/MS)
R.T.=0.82; [M+H].sup.+=290.3.
Example 24
(R)--N-(6-Methoxy-[1,2,4]-triazolo[1,5-a]pyridin-2-yl)-4H-1'-azaspiro[oxaz-
ole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00498##
[0282] Step A: 5-Methoxypyridin-2-amine ethoxycarbonyl thiourea
##STR00499##
[0284] To 5-methoxypyridin-2-amine (5 g, 40 mmol) in dioxane (40
mL) was added ethoxycarbonyl isothiocyanate (5.23 mL, 44.3 mmol).
The reaction mixture was stirred at room temperature overnight. The
mixture was concentrated in vacuo to afford
5-methoxypyridin-2-amine ethoxycarbonyl thiourea (10.28 g, 40.3
mmol, 100% yield). .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm
12.03 (1H, br. s.), 11.37 (1H, br. s.), 8.53 (1H, br. s.), 8.11
(1H, d, J=2.93 Hz), 7.50 (1H, dd, J=8.97, 3.11 Hz), 4.22 (2H, q,
J=7.07 Hz), 3.84 (3H, s), 1.26 (3H, t, J=7.14 Hz). MS (LC/MS)
R.T.=2.40; [M+H].sup.+=256.1.
Step B: 6-Methoxy-[1,2,4]-triazolo[1,5-a]pyridin-2-amine
##STR00500##
[0286] To 5-methoxypyridin-2-amine ethoxycarbonyl thiourea (10.21
g, 40 mmol) in ethanol (57 mL) and methanol (57 mL) was added
hydroxylamine hydrochloride (14 g, 200 mmol) and Hunig's Base (21
mL, 120 mmol). The mixture was stirred at room temperature for 2
hours, and then heated at 60.degree. C. for 4 hours. The reaction
was cooled to room temperature and filtered to remove solids. It
was concentrated in vacuo and then suspended in chloroform. The
solids were filtered off to yield
6-methoxy-[1,2,4]triazolo[1,5-a]pyridin-2-amine (6.05 g, 33.2 mmol,
83% yield). .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm 8.30
(1H, d, J=1.83 Hz), 7.24-7.32 (1H, m), 7.16-7.23 (1H, m), 5.82 (2H,
br. s.), 3.78 (3H, s). MS (LC/MS) R.T.=0.53; [M+H].sup.+=165.2.
[0287] Two grams of 6-methoxy-[1,2,4]triazolo[1,5-a]pyridin-2-amine
were purified via flash chromatography (2-20% [10% ammonium
hydroxide/methanol]/chloroform) to afford
6-methoxy-[1,2,4]triazolo[1,5-a]pyridin-2-amine (1.35 g, 8.22 mmol,
67.5% yield). .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm 8.29
(1H, d, J=2.20 Hz), 7.24-7.30 (1H, m), 7.15-7.22 (1H, m), 5.79 (2H,
s), 3.78 (3H, s).
Step C:
2-Isothiocyanato-6-methoxy-[1,2,4]triazolo[1,5-a]pyridine
##STR00501##
[0289] To 6-methoxy-[1,2,4]triazolo[1,5-a]pyridin-2-amine (0.3 g,
1.8 mmol) in dichloromethane (15 mL) was added
1,1'-thiocarbonyldi-2(1H)-pyridone (0.51 g, 2.2 mmol). The reaction
was stirred at room temperature overnight, concentrated in vacuo
and purified via flash chromatography yielding
2-isothiocyanato-6-methoxy-[1,2,4]triazolo[1,5-a]pyridine (166 mg,
0.8 mmol, 44% yield) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. ppm 8.03 (1H, d, J=2.20 Hz), 7.54 (1H, d,
J=9.88 Hz), 7.34 (1H, dd, J=9.51, 2.56 Hz), 3.88 (3H, s).
Step D:
(R)--N-(6-Methoxy-[1,2,4]-triazolo[1,5-a]pyridin-2-yl)-4H-1'-azasp-
iro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00502##
[0291] To 2-isothiocyanato-6-methoxy-[1,2,4]triazolo[1,5-a]pyridine
(160 mg, 0.78 mmol) in N,N-dimethylformamide (5 ml) was added
(S)-3-(aminomethyl)quinuclidin-3-ol, 2HCl (178 mg, 0.78 mmol) and
triethylamine (0.32 ml, 2.33 mmol). The reaction was stirred at
70.degree. C. for 1 hour. N,N'-Diisopropylcarbodiimide (0.36 ml,
2.33 mmol) was then added to the reaction mixture. The mixture was
heated at 70.degree. C. for 4 hours, then cooled and poured into
aqueous sodium bicarbonate/chloroform. The product was extracted
(3.times.) with chloroform. The combined organics were washed with
water (3.times.), dried over sodium sulfate, filtered, and
concentrated in vacuo, then purified via flash chromatography
(2-20% [10% ammonium hydroxide:methanol]/chloroform) to yield
(R)--N-(6-methoxy-[1,2,4]triazolo[1,5-a]pyridin-2-yl)-4H-1'-azaspiro-[oxa-
zole-5,3'-bicyclo[2.2.2]octan]-2-amine (114.5 mg, 0.33 mmol, 42%
yield) as a white powder. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
ppm 8.51 (1H, br. s.), 8.08 (1H, d, J=2.44 Hz), 7.35 (1H, d, J=9.77
Hz), 7.18 (1H, dd, J=9.46, 2.44 Hz), 3.92 (1H, d, J=8.85 Hz), 3.84
(3H, s), 3.58 (1H, d, J=8.85 Hz), 3.36-3.41 (1H, m), 2.72-3.05 (4H,
m), 2.18-2.26 (1H, m, J=13.26, 9.98, 3.66, 3.49, 3.49 Hz), 2.13
(1H, br. s.), 1.79 (1H, br. s.), 1.67-1.76 (1H, m, J=13.96, 9.84,
4.27, 4.27 Hz), 1.45-1.63 (2H, m). MS (LC/MS) R.T.=0.86;
[M+H].sup.+=329.2.
Example 25
(R)--N-(5-(Trifluoromethyl)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00503##
[0292] Step A: 5-(Trifluoromethyl)pyrazin-2-amine
##STR00504##
[0294] To an ice bath-cooled solution of 5,6-diaminopyrimidin-4-ol
(18 g, 143 mmol) in 3M sodium hydroxide (180 mL, 540 mmol), was
added 3,3-dibromo-1,1,1-trifluoropropan-2-one (25.2 g, 93 mmol).
The reaction was stirred for 3 days at ambient temperature. The
solids were filtered, dissolved in 60% sulfuric acid (140 mL), and
stirred at 135.degree. C. for 8 h. The reaction was cooled, poured
over ice and allowed to sit for 16 hours. The solution was
neutralized to pH 8 with conc. ammonium hydroxide and extracted
with ethyl acetate (5.times.100 mL), dried over sodium sulfate,
filtered and concentrated. The solid residue was recrystallized
from benzene/hexane to afford 5-(trifluoromethyl)pyrazin-2-amine
(2.28 g, 14 mmol, 15% yield). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 8.32 (1H, s), 7.99 (1H, d, J=1.26 Hz), 5.02 (2H, br.
s.). MS (LC/MS) R.T.=1.56; [M+H].sup.+=164.03.
Step B:
(R)--N-(5-(Trifluoromethyl)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine
##STR00505##
[0296]
(R)--N-(5-(trifluoromethyl)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine was prepared by following the
general procedures of Example 23, Steps A-B and using
5-(trifluoromethyl)pyrazin-2-amine (from Step A above) as the
starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm
9.08 (1H, br. s.), 8.35 (1H, s), 8.32 (1H, s), 3.95 (1H, d, J=9.57
Hz), 3.61 (1H, d, J=9.57 Hz), 3.30 (1H, dd, J=14.86, 1.76 Hz),
2.65-2.99 (5H, m), 2.05-2.16 (2H, m), 1.64-1.74 (1 H, m), 1.42-1.57
(2H, m). MS (LC/MS) R.T.=1.06; [M+H].sup.+=328.30.
Example 26
(R)--N-(6-Fluoro-1H-indazol-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00506##
[0297] Step A: 6-Fluoro-1H-indazol-3-amine
##STR00507##
[0299] To 2,4-difluorobenzonitrile (1.21 g, 8.70 mmol) was added
hydrazine monohydrate (8.46 mL, 174 mmol). The mixture was heated
to reflux for 5 hours and then poured onto ice. The solution was
extracted with ethyl acetate, dried with magnesium sulfate,
filtered and concentrated. The residue was purified by column
chromatography (25-100% ethyl acetate/hexane) to afford
6-fluoro-1H-indazol-3-amine (0.5 g, 3.3 mmol, 38% yield) as light
yellow powder. .sup.1H NMR (500 MHz, DMSO-D6) .delta. ppm 11.42 (s,
1H), 7.70 (dd, J=8.55, 5.49 Hz, 1H), 6.97 (dd, J=10.07, 1.83 Hz,
1H), 6.72-6.79 (m, 1H), 5.40 (s, 2H). MS (LC/MS) R.T.=0.61;
[M+H].sup.+=152.11.
Step B: 6-Fluoro-3-isothiocyanato-1H-indazole
##STR00508##
[0301] To 6-fluoro-1H-indazol-3-amine (0.32 g, 2.1 mmol) in
dichloromethane (15 mL) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (0.53 g, 2.30 mmol). The
reaction was stirred at 50.degree. C. for 3 hours. The reaction was
cooled to room temperature and the crude product was purified by
column chromatography (25% ethyl acetate/hexanes) to afford
6-fluoro-3-isothiocyanato-1H-indazole (0.30 g, 1.53 mmol, 73.0%
yield) as a light yellow powder. .sup.1H NMR (500 MHz, DMSO-D6)
.delta. ppm 13.39 (s, 1H), 7.78 (dd, J=8.85, 4.88 Hz, 1H), 7.41
(dd, J=9.31, 1.68 Hz, 1H), 7.14 (td, J=9.16, 1.83 Hz, 1H). MS
(LC/MS) R.T.=3.69; [M+H].sup.+=194.07.
Step C:
(R)--N-(6-Fluoro-1H-indazol-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00509##
[0303] To 6-fluoro-3-isothiocyanato-1H-indazole (0.20 g, 1.04 mmol)
in DMF (15 mL) was added triethylamine (0.43 mL, 3.11 mmol) and
(S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.26 g, 1.14
mmol) at room temperature. The reaction was stirred at 70.degree.
C. for 2 hours. The reaction was cooled to room temperature and
treated with N,N'-diisopropylcarbodiimide (0.48 mL, 3.11 mmol). The
reaction was then heated to 70.degree. C. for 2 hours. The reaction
was cooled to ambient temperature and concentrated. The crude
product was purified by column chromatography (85% chloroform, 14%
methanol, 1% ammonium hydroxide) to afford
(R)--N-(6-fluoro-1H-indazol-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine (0.22 g, 0.66 mmol, 64% yield) as a white
powder. .sup.1H NMR (500 MHz, DMSO-D6) .delta. ppm 12.16 (s, 1H),
8.00 (s, 1H), 7.59-7.67 (m, 1 H), 7.09 (dd, J=9.77, 2.14 Hz, 1H),
6.81-6.88 (m, 1H), 3.81 (d, J=9.16 Hz, 1H), 3.56 (d, J=8.85 Hz,
1H), 3.00 (s, 2H), 2.78 (s, 2H), 2.67 (t, J=7.32 Hz, 2H), 2.01 (d,
J=2.44 Hz, 1H), 1.92 (s, 1H), 1.59 (d, J=5.80 Hz, 2H), 1.46 (s,
1H). MS (LC/MS) R.T.=1.44; [M+H].sup.+=316.16.
Example 27
(R)--N-(Furo[3,2-c]pyridin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00510##
[0304] Step A: Furo[3,2-c]pyridin-4-amine
##STR00511##
[0306] To 4-chlorofuro[3,2-c]pyridine (1 g, 6.5 mmol) in toluene
under nitrogen was added racemic BINAP (0.243 g, 0.4 mmol),
Pd.sub.2(dba).sub.3 (0.12 g, 0.13 mmol) and sodium tert-butoxide
(0.88 g, 9.1 mmol). Benzophenoneimine (1.3 mL, 7.81 mmol) was added
and the mixture was heated to 80.degree. C. for 3 h and cooled to
room temperature. The reaction mixture was diluted with ether,
filtered through Celite, and washed with ether. The filtrate was
concentrated and the deep orange residue was taken up in methanol
(90 ml) and treated with hydroxylamine (1.2 mL, 19.5 mmol). The
mixture was stirred at ambient temperature for 18 h and
concentrated. The residue was purified by column chromatography
(95-100% ethyl acetate/hexanes) to afford
furo[3,2-c]pyridin-4-amine (776 mg, 5.79 mmol, 89% yield) as a deep
orange solid. MS (LC/MS) R.T.=0.51; [M+H].sup.+=135.02.
Step B:
(R)--N-(Furo[3,2-c]pyridin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00512##
[0308]
(R)--N-(Furo[3,2-c]pyridin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine was prepared by following the general
procedures of Example 23, Steps A-B using
furo[3,2-c]pyridin-4-amine (Step A above) as the starting material.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.08 (1H, d, J=5.79 Hz),
7.69 (1H, d, J=2.01 Hz), 6.99-7.13 (2H, m), 4.00 (1H, d, J=10.07
Hz), 3.69 (1H, d, J=10.07 Hz), 3.19-3.26 (1H, m), 3.05-3.13 (1H,
m), 2.93 (2H, t, J=7.43 Hz), 2.73-2.87 (2H, m), 2.08-2.24 (2H, m),
1.52-1.82 (3H, m). MS (LC/MS) R.T.=0.68; [M+H].sup.+=299.19.
Example 28
(R)--N-(5-Phenylpyridin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oc-
tan]-2-amine
##STR00513##
[0309] Step A: 5-Phenylpyridin-3-amine
##STR00514##
[0311] A mixture of 5-bromopyridin-3-amine (248 mg, 1.43 mmol),
Pd(PPh.sub.3).sub.4 (50.4 mg, 0.04 mmol), toluene (3 mL), sodium
carbonate (2 M, 3 mL, 6 mmol), and phenylboronic acid (195 mg, 1.60
mmol) dissolved in ethanol (3 mL) was heated for 4 hours in an oil
bath at 90.degree. C. and allowed to cool to room temperature for
16 hours. The reaction mixture was transferred to a separatory
funnel and partitioned between ethyl acetate and water. The aqueous
phase was washed once more with ethyl acetate, and the combined
organic phases were washed with brine and dried over magnesium
sulfate, filtered and concentrated in vacuo. The residue was
purified by column chromatography (80% ethyl acetate/hexanes) to
afford 5-phenylpyridin-3-amine (31.9 mg, 0.19 mmol, 13% yield) as a
white solid. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm
8.17-8.42 (m, 1H), 8.02-8.20 (m, 1H), 7.32-7.62 (m, 4H), 7.25 (s,
1H), 7.06-7.20 (m, 1H). MS (LC/MS) R.T.=0.91;
[M+H].sup.+=171.09.
Step B:
(R)--N-(5-Phenylpyridin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine
##STR00515##
[0313]
(R)--N-(5-Phenylpyridin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine was prepared by following the general
procedures of Example 23, Steps A-B using 5-phenylpyridin-3-amine
(from Step A above) as the starting material. .sup.1H NMR (500 MHz,
MeOD-d.sub.4) .delta. ppm 8.40 (br. s., 2H), 7.66 (d, J=7.32 Hz,
2H), 7.46-7.56 (m, 3H), 7.43 (d, J=7.32 Hz, 1H), 3.79-4.02 (m, 1H),
3.51-3.68 (m, 1H), 3.22 (d, J=14.95 Hz, 1H), 3.02-3.15 (m, 1H),
2.72-2.99 (m, 3H), 2.14 (br. s., 2H), 1.76 (dd, J=9.31, 4.12 Hz,
3H), 1.12-1.35 (m, 1H). MS (LC/MS) R.T.=0.90;
[M+H].sup.+=335.17.
Example 29
(R)--N-(2-Phenylpyridin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oc-
tan]-2-amine
##STR00516##
[0315]
(R)--N-(2-Phenylpyridin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine was prepared from 2-bromopyridin-4-amine by
following the general procedures of Example 28, Steps A-B. .sup.1H
NMR (500 MHz, MeOD) .delta. ppm 8.38 (d, J=5.49 Hz, 1H), 7.88 (d,
J=7.93 Hz, 2H), 7.71-7.84 (m, 1H), 7.48 (d, J=7.63 Hz, 3H),
7.19-7.36 (m, 1H), 3.94-4.09 (m, 1H), 3.61-3.79 (m, 1H), 3.17-3.27
(m, 1H), 3.00-3.14 (m, 1H), 2.74-3.00 (m, 4H), 2.05-2.23 (m, 2H),
1.55-1.86 (m, 3H). MS (LC/MS) R.T.=0.86; [M+H].sup.+=335.23.
Example 30
(R)--N-(6-Phenylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oc-
tan]-2-amine
##STR00517##
[0317]
(R)--N-(6-Phenylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine was prepared from 6-bromopyridin-2-amine by
following the general procedures of Example 28, Steps A-B. .sup.1H
NMR (500 MHz, MeOD-d.sub.4) .delta. ppm 7.91 (d, J=7.63 Hz, 2H),
7.71 (t, J=7.78 Hz, 1H), 7.32-7.55 (m, 5 H), 4.04 (d, J=10.07 Hz,
1H), 3.72 (d, J=9.77 Hz, 1H), 3.23 (d, J=1.22 Hz, 1H), 3.12 (s,
1H), 2.95 (s, 2H), 2.84 (s, 2H), 2.16 (br. s., 2H), 1.58-1.85 (m,
3H). MS (LC/MS) R.T.=0.75; [M+H].sup.+=335.23.
Example 31
(R)--N-(6-(Difluoromethoxy)-1-methyl-1H-indazol-3-yl)-4H-1'-azaspiro[oxazo-
le-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00518##
[0318] Step A: 6-(Difluoromethoxy)-1-methyl-1H-indazol-3-amine
##STR00519##
[0320] To 4-(difluoromethoxy)-2-fluorobenzonitrile (1 g, 5.3 mmol)
was added methylhydrazine (4.92 g, 107 mmol). The mixture was
heated to 50.degree. C. for 5 hours and then cooled to room
temperature. The crude product was purified by column
chromatography (40-100% ethyl acetate/hexanes) to afford
6-(difluoromethoxy)-1-methyl-1H-indazol-3-amine (0.5 g, 2.35 mmol,
44% yield) as a white powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 7.70 (d, J=8.55 Hz, 1H), 7.14 (s, 1H), 6.73 (d, J=8.55
Hz, 1H), 5.49 (s, 1H), 3.70 (s, 3H).
Step B:
(R)--N-(6-(Difluoromethoxy)-1-methyl-1H-indazol-3-yl)-4H-1'-azaspi-
ro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00520##
[0322]
(R)--N-(6-(difluoromethoxy)-1-methyl-1H-indazol-3-yl)-4H-1'-azaspir-
o[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared by
following the general procedures of Example 23, Steps A-B using
6-(difluoromethoxy)-1-methyl-1H-indazol-3-amine (Step A) as the
starting material. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
7.64 (d, J=8.55 Hz, 1H), 7.28 (d, J=5.19 Hz, 1H), 6.83 (dd, J=8.55,
1.83 Hz, 1H), 3.88 (s, 3H), 3.77-3.84 (m, 1H), 3.51-3.64 (m, 1H),
3.00 (s, 2H), 2.72-2.84 (m, 2H), 2.67 (t, J=7.48 Hz, 2H), 2.02 (br.
s., 1H), 1.85-1.98 (m, 1H), 1.59 (d, J=5.80 Hz, 2H), 1.36-1.53 (m,
1H), 1.09 (d, J=6.41 Hz, 1H). MS (LC/MS) R.T.=1.04;
[M+H].sup.+=378.19.
Example 32
(R)--N-(5-(Difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxa-
zole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00521##
[0323] Step A: 2-(Difluoromethoxy)-5-nitropyridine
##STR00522##
[0325] To 2-hydroxy-5-nitropyridine (7 g, 50 mmol) in acetonitrile
(500 mL) was added sodium sulfate (1.5 g, 10.6 mmol), and
2,2-difluoro-2-(fluorosulfonyl)acetic acid (6.2 mL, 60 mmol) and
the reaction was allowed to stir at room temperature for 16 hours.
The reaction was quenched with saturated aqueous sodium bicarbonate
and the acetonitrile was removed in vacuo. The remaining aqueous
component was extracted with ethyl acetate, washed with brine,
dried over sodium sulfate, filtered and concentrated in vacuo. The
pale brown oily solid was triturated with ether/hexanes, filtered
and the filtrate concentrated to afford
2-(difluoromethoxy)-5-nitropyridine (4.7 g, 24.7 mmol, 49% yield)
as a yellow oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
9.14 (d, J=2.76 Hz, 1H), 8.68 (dd, J=9.03, 2.76 Hz, 1H), 7.98 (s,
0.5H), 7.62 (s, 0.5H), 7.34 (d, J=9.03 Hz, 1H).
Step B: 6-(Difluoromethoxy)pyridin-3-amine
##STR00523##
[0327] To 2-(difluoromethoxy)-5-nitropyridine (4.7 g, 24.7 mmol) in
degassed methanol (100 mL) was added 10% palladium on carbon (500
mg, 0.47 mmol) and the reaction was hydrogenated at atmospheric
pressure for 1 hour. To this was added acetic acid (2.83 mL, 49.4
mmol) and the reaction was filtered through Celite and concentrated
in vacuo to afford 6-(difluoromethoxy)pyridin-3-amine (6.33 g, 25.9
mmol, 105% yield) as an olive green liquid. .sup.1H NMR (400 MHz,
MeOD-d.sub.4) .delta. ppm 7.60 (d, J=2.76 Hz, 1H), 7.37 (s, 0.5H),
7.15 (dd, J=8.66, 2.89 Hz, 1H), 7.00 (s, 0.5H), 6.71 (d, J=8.78 Hz,
1H).
Step C: 5-(Difluoromethoxy)thiazolo[5,4-b]pyridin-2-amine
##STR00524##
[0329] To acetic acid (10 mL) cooled in an ice bath was added
potassium thiocyanate (3.18 g, 32.8 mmol) and
6-(difluoromethoxy)pyridin-3-amine (1 g, 4.1 mmol). The reaction
was cooled in an ice-salt bath until the reaction temperature
reached <0.degree. C. A solution of bromine (0.65 mL, 12.7 mmol)
in acetic acid (3 mL) was added dropwise over 2 hours at a rate
that maintained the reaction temperature <0.degree. C. This gave
a very thick mixture. After the addition was complete, the mixture
was left to stir and allowed to slowly warm to room temperature
overnight. After stirring overnight, water (5 mL) was added and the
mixture was heated to 85.degree. C. in an oil bath. This mixture
was then filtered while still hot. The yellow filter cake was
returned to the reaction flask, and an additional 5 mL acetic acid
was added. The mixture was heated again to 85.degree. C., and then
filtered while still hot. The combined filtrates were cooled in an
ice bath and neutralized to pH 8 with concentrated ammonium
hydroxide. A yellow precipitate formed which was then collected by
filtration. This crude material was purified by column
chromatography (12-100% ethyl acetate/hexanes) to afford
5-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-amine (321 mg, 1.48
mmol, 36.1% yield) as a yellow solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 7.64-7.81 (m, 2H), 6.92 (d, J=8.53 Hz,
1H). MS (LC/MS) R.T.=1.66; [M+H].sup.+=218.10.
Step D:
N-(5-(Difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)-1H-imidazole-1--
carbothioamide
##STR00525##
[0331] 5-(Difluoromethoxy)thiazolo[5,4-b]pyridin-2-amine (310 mg,
1.43 mmol) and di(1H-imidazol-1-yl)methanethione (311 mg, 1.75
mmol) were dissolved in acetonitrile (5 mL) and heated to
70.degree. C. in a sealed vial for 10 hours. The vial was then
stored in the freezer for 16 hours. The solids were collected by
filtration and washed with acetonitrile to afford
N-(5-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)-1H-imidazole-1--
carbothioamide (296 mg, 0.72 mmol, 51% yield). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 8.77 (d, J=4.77 Hz, 1H), 8.06 (d,
J=8.53 Hz, 1H), 8.01 (s, 1 H), 7.72 (s, 1H), 7.54 (s, 1H),
6.94-7.13 (m, 1H).
Step E:
(R)--N-(5-(Difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)-4H-1'-azas-
piro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00526##
[0333] To
N-(5-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)-1H-imidazole--
1-carbothioamide (296 mg, 0.72 mmol) in N,N-dimethylformamide (4
mL) in a sealed vial was added (S)-3-(aminomethyl)quinuclidin-3-ol,
2HCl (175 mg, 0.76 mmol) and triethylamine (0.3 ml, 2.2 mmol) and
the mixture was heated to 70.degree. C. overnight.
[0334] To this was added N,N'-diisopropylcarbodiimide (350 .mu.l,
2.25 mmol) and the reaction was heated to 70.degree. C. for 6
hours. It was cooled to ambient temperature and poured into
water/chloroform, extracted with additional chloroform and
concentrated in vacuo. The residue was then taken up in toluene and
washed with water to remove the residual N,N-dimethylformamide. The
residue was purified by column chromatography (2%-20% (10% ammonium
hydroxide/methanol)/chloroform) to afford
(R)--N-(5-(difluoromethoxy)thiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[ox-
azole-5,3'-bicyclo[2.2.2]octan]-2-amine (104 mg, 0.27 mmol, 37%
yield) as a pale cream colored solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 7.99 (d, J=8.78 Hz, 1H), 7.68 (s, 1H),
7.03 (d, J=8.53 Hz, 1H), 3.87 (d, J=10.04 Hz, 1H), 3.62 (d, J=10.04
Hz, 1H), 3.03 (d, J=5.02 Hz, 2H), 2.80 (d, J=9.03 Hz, 2H), 2.65 (t,
J=7.65 Hz, 2H), 2.08 (br. s., 1H), 1.83-1.99 (m, 1H), 1.43-1.68 (m,
3H). MS (LC/MS) R.T.=1.73; [M+H].sup.+=382.20.
Example 33
(R)--N-(6-Isopropoxybenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00527##
[0336]
(R)--N-(6-Isopropoxybenzo[d]thiazol-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine was prepared from
4-isopropoxyaniline by following the general procedures of Example
32, Steps C-E. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.49
(d, J=8.78 Hz, 1H), 7.38 (d, J=2.51 Hz, 1H), 6.90 (dd, J=8.78, 2.51
Hz, 1H), 4.50-4.68 (m, 1H), 3.87 (d, J=10.04 Hz, 1H), 3.62 (d,
J=9.79 Hz, 1H), 2.79 (d, J=9.03 Hz, 2H), 2.66 (t, J=7.78 Hz, 2H),
2.06 (br. s., 1H), 1.85-1.96 (m, 1H), 1.42-1.67 (m, 3H), 1.17-1.34
(m, 6H), 0.96-1.19 (m, 2H).
Example 34
(R)--N-(5-(Pyrrolidin-1-yl)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00528##
[0337] Step A: 5-(Pyrrolidin-1-yl)pyrazin-2-amine
##STR00529##
[0339] A mixture of 5-bromopyrazin-2-amine (1.2 g, 6.9 mmol) and
pyrrolidine (4 mL, 48 mmol) was microwaved at 180.degree. C., 200 W
for 2 h. The reaction was diluted into 125 mL ethyl acetate and
extracted with water (3.times.50 mL) and brine (50 mL). It was
dried over sodium sulfate, filtered and concentrated. The crude
product was purified by column chromatography (0 to 3%
methanol/methylene chloride) to afford
5-(pyrrolidin-1-yl)pyrazin-2-amine (495 mg, 3. mmol, 43.7% yield).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.52 (1H, d, J=1.51
Hz), 7.36 (1H, d, J=1.76 Hz), 5.21 (2 H, s), 3.24-3.29 (4H, m),
1.90 (4H, ddd, J=6.48, 3.53, 3.34 Hz). MS (LC/MS) R.T.=0.52;
[M+H].sup.+=165.29.
Step B:
(R)--N-(5-(Pyrrolidin-1-yl)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine
##STR00530##
[0341]
(R)--N-(5-(Pyrrolidin-1-yl)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine was prepared from
5-(pyrrolidin-1-yl)pyrazin-2-amine by following the general
procedures of Example 23, Steps A-B. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 8.51 (1H, br. s.), 8.02 (1H, s), 7.44 (1H,
d, J=0.92 Hz), 3.84 (1 H, d, J=9.16 Hz), 3.51 (1H, d, J=8.85 Hz),
3.39-3.45 (4H, m), 3.31 (1H, dd, J=14.80, 1.07 Hz), 2.69-3.04 (5H,
m), 2.15-2.25 (1H, m), 2.09 (1H, br. s.), 1.98-2.02 (4H+ HOD, m),
1.64-1.74 (1H, m), 1.42-1.61 (2H, m). MS (LC/MS) R.T.=0.76;
[M+H].sup.+=329.40.
Example 35
(R)-1-(5-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)pyra-
zin-2-yl)pyrrolidin-2-one
##STR00531##
[0342] Step A: 1-(5-Aminopyrazin-2-yl)pyrrolidin-2-one
##STR00532##
[0344] A mixture of 5-bromopyrazin-2-amine (5 g, 29 mmol),
pyrrolidin-2-one (11 mL, 144 mmol), copper (I) iodide (1.1 g, 5.75
mmol), potassium carbonate (7.94 g, 57.5 mmol), and
(1R,2R)-cyclohexane-1,2-diamine (1.38 mL, 11.49 mmol) was refluxed
under nitrogen in dioxane (100 mL) for 18 h. After cooling, 200 mL
ethyl acetate and 20 mL methanol were added to the reaction. This
was filtered through celite, concentrated, and absorbed onto sodium
sulfate for purification by column chromatography (0-5%
methanol/methylene chloride). The purified product was
recrystallized from ether/ethyl acetate to afford
1-(5-aminopyrazin-2-yl)pyrrolidin-2-one (1.57 g, 8.81 mmol, 30.7%
yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 8.74 (1H,
d, J=1.51 Hz), 7.69 (1H, d, J=1.51 Hz), 6.20 (2H, s), 3.84 (2H, t,
J=7.05 Hz), 2.49 (2H, t, J=7.93 Hz), 2.04 (2H, dq, J=7.68, 7.51
Hz). MS (LC/MS) R.T.=0.48; [M+H].sup.+=179.27.
Step B:
(R)-1-(5-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylam-
ino)pyrazin-2-yl)pyrrolidin-2-one
##STR00533##
[0346]
(R)-1-(5-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylami-
no)pyrazin-2-yl)pyrrolidin-2-one was prepared from
1-(5-Aminopyrazin-2-yl)pyrrolidin-2-one by following the general
procedures of Example 23, Steps A-B. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 9.20 (1H, d, J=1.22 Hz), 8.91 (1H, br. s.),
8.13 (1H, s), 4.02 (2H, t, J=7.02 Hz), 3.92 (1H, d, J=9.46 Hz),
3.58 (1H, d, J=9.16 Hz), 3.33 (1H, dd, J=14.95, 1.53 Hz), 2.71-3.01
(5H, m), 2.63 (2H, t, J=8.09 Hz), 2.08-2.23 (4H, m), 1.66-1.76 (1H,
m), 1.42-1.61 (2H, m). MS (LC/MS) R.T.=0.67;
[M+H].sup.+=343.30.
Example 36
(R)--N-(5-(Pyridin-3-yl)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine
##STR00534##
[0347] Step A: 5-(Pyridin-3-yl)pyrazin-2-amine
##STR00535##
[0349] Pyridin-3-ylboronic acid (307 mg, 2.50 mmol),
5-bromopyrazin-2-amine (391 mg, 2.25 mmol) and
dichlorobis(triphenylphosphine)-palladium(II) (88 mg, 0.13 mmol)
were added to degassed dioxane (12 mL) and the mixture was stirred
for 30 min. Then sodium carbonate (795 mg, 7.50 mmol) and degassed
water (8 mL) were added and the reaction was heated in a closed
reaction vial at 100.degree. C. for 8 h. The reaction was allowed
to stand at ambient temperature over the weekend. It was diluted
into ethyl acetate (100 mL) and extracted with brine (3.times.25
mL). The organic layers were dried over sodium sulfate and
concentrated in vacuo. The crude product was purified by column
chromatography (0 to 5% methanol/ethyl acetate) to afford
5-(pyridin-3-yl)pyrazin-2-amine (235 mg, 1.37 mmol, 54.6% yield).
.sup.1H NMR (400 MHz, Acetone) .delta. ppm 9.13 (1H, d, J=1.51 Hz),
8.54 (1H, d, J=1.26 Hz), 8.50 (1H, dd, J=4.78, 1.51 Hz), 8.22-8.28
(1H, m), 8.07 (1H, d, J=1.26 Hz), 7.39 (1H, ddd, J=7.87, 4.72, 0.76
Hz), 6.05 (2H, br. s.). MS (LC/MS) R.T.=0.58;
[M+H].sup.+=173.20.
Step B:
(R)--N-(5-(Pyridin-3-yl)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine
##STR00536##
[0351]
(R)--N-(5-(Pyridin-3-yl)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine was prepared from
5-(pyridin-3-yl)pyrazin-2-amine by following the general procedures
of Example 23, Steps A-B. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 9.12 (1H, dd, J=2.27, 0.76 Hz), 9.08 (1H, br. s.), 8.59 (1H,
dd, J=4.78, 1.76 Hz), 8.55 (1H, d, J=1.51 Hz), 8.45 (1H, d, J=1.26
Hz), 8.25 (1H, dt, J=7.99, 1.92 Hz), 7.38 (1H, ddd, J=8.06, 4.78,
0.76 Hz), 3.97 (1H, d, J=9.57 Hz), 3.63 (1H, d, J=9.32 Hz), 3.36
(1H, dd, J=14.86, 1.76 Hz), 2.69-3.06 (5H, m), 2.14-2.24 (1H, m),
2.12 (1H, br. s.), 1.66-1.77 (1H, m, J=13.94, 9.66, 4.31, 4.31 Hz),
1.44-1.62 (2H, m). MS (LC/MS) R.T.=0.65; [M+H].sup.+=337.30.
Example 37
(R)--N-(5-(6-Methoxypyridin-3-yl)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octan]-2-amine
##STR00537##
[0353]
(R)--N-(5-(6-Methoxypyridin-3-yl)pyrazin-2-yl)-4H-1'-azaspiro[oxazo-
le-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared from
5-bromopyrazin-2-amine by following the general procedures of
Example 36, Steps A-B. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 9.04 (1H, br. s.), 8.66 (1H, d, J=2.01 Hz), 8.46 (1H, d, J=1.51
Hz), 8.41 (1H, d, J=1.01 Hz), 8.15 (1H, dd, J=8.81, 2.52 Hz), 6.82
(1H, d, J=8.31 Hz), 3.97 (3H, s), 3.95 (1H, d, J=9.57 Hz), 3.61
(1H, d, J=9.32 Hz), 3.35 (1H, dd, J=14.86, 1.51 Hz), 2.68-3.07 (5H,
m), 2.14-2.24 (1H, m, J=13.27, 9.93, 3.53, 3.38, 3.38 Hz), 2.11
(1H, br. s.), 1.67-1.77 (1H, m), 1.45-1.61 (2H, m). MS (LC/MS)
R.T.=0.81; [M+H].sup.+=367.40.
Example 38
(R)--N-(6-Methoxyquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00538##
[0354] Step A: 7-Methoxyquinoxalin-2(1H)-one and
6-methoxyquinoxalin-2(1H)-one
##STR00539##
[0356] A 50% solution of ethyl 2-oxoacetate (18.47 mL, 93 mmol) in
toluene was added to a solution of 4-methoxybenzene-1,2-diamine
(10.73 g, 78 mmol) in ethanol (100 mL) at ambient temperature and
the reaction was refluxed for 2 h. The reaction was concentrated in
vacuo and crystallized from ethanol to afford a mixture of
6-methoxyquinoxalin-2(1H)-one and 7-methoxyquinoxalin-2(1H)-one
(5.73 g, 32.50 mmol, 42% yield). MS (LC/MS) R.T.=0.68;
[M+H].sup.+=177.10.
Step B: 2-Chloro-6-methoxyquinoxaline
##STR00540##
[0358] A mixture of 6-methoxyquinoxalin-2(1H)-one and
7-methoxyquinoxalin-2(1H)-one (5.67 g, 32.20 mmol) was refluxed in
phosphorus oxychloride (120 mL) for 1 h. The reaction was
concentrated and quenched by addition of ice, then basified with
sodium carbonate, and extracted with ethyl acetate (3.times.200
mL). The organic layers were combined and concentrated in vacuo.
The crude product was absorbed onto sodium sulfate and purified by
column chromatography (0 to 5% ethyl acetate/hexanes) to afford
2-chloro-6-methoxyquinoxaline (2.21 g, 11.36 mmol, 35% yield).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.69 (s, 1H), 7.88
(d, J=9.32 Hz, 1H), 7.43 (dd, J=9.32, 2.77 Hz, 1H), 7.37 (d, J=2.77
Hz, 1H), 3.95 (s, 3H).
Step C: N-(2,4-Dimethoxybenzyl)-6-methoxyquinoxalin-2-amine
##STR00541##
[0360] 2-Chloro-6-methoxyquinoxaline (0.93 g, 4.77 mmol) and
(2,4-dimethoxyphenyl)methanamine (2.2 ml, 14.64 mmol) were
microwaved in dimethylsulfoxide (5 mL) for 30 min at 150.degree. C.
The reaction was diluted into ethyl acetate (250 mL) and extracted
with brine (3.times.100 mL). The crude product was purified by
column chromatography (20 to 80% ethyl acetate/hexanes) to afford
N-(2,4-dimethoxybenzyl)-6-methoxyquinoxalin-2-amine (1.46 g, 87%
yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.13 (1H, s),
7.59-7.63 (1H, m), 7.30 (1H, d, J=8.31 Hz), 7.21-7.24 (2H, m), 6.47
(1H, d, J=2.27 Hz), 6.42 (1H, dd, J=8.31, 2.27 Hz), 5.10 (1H, t,
J=5.92 Hz), 4.61 (2H, d, J=5.79 Hz), 3.88 (3H, s), 3.84 (3H, s),
3.78 (3 H, s). MS (LC/MS) R.T.=1.95; [M+H].sup.+=326.23.
Step D: 6-Methoxyquinoxalin-2-amine
##STR00542##
[0362] N-(2,4-Dimethoxybenzyl)-6-methoxyquinoxalin-2-amine (2.8 g,
8.61 mmol) was stirred in TFA (10 mL, 130 mmol) and dichloromethane
(10 mL) at ambient temperature for 30 min. The solvents were
removed in vacuo. Saturated aq. sodium hydrogen carbonate (200 mL)
was added to the red residue, which then precipitated a yellow
solid. The mixture was extracted extensively with dichloromethane.
The organic layer was dried over sodium sulfate, filtered and
concentrated in vacuo to afford 6-methoxyquinoxalin-2-amine (1.50
g, 8.56 mmol, 99% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 8.27 (1H, s), 7.54-7.58 (1H, m), 7.25-7.29 (2H, m), 4.71 (2H,
br. s.), 3.90 (3H, s). MS (LC/MS) R.T.=0.86;
[M+H].sup.+=176.23.
Step E:
(R)--N-(6-Methoxyquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00543##
[0364]
(R)--N-(6-Methoxyquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine was prepared from
6-methoxyquinoxalin-2-amine by following the general procedures of
Example 23, Steps A-B. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 9.61 (1H, br. s.), 8.57 (1H, s), 7.62 (1H, d, J=9.06 Hz),
7.23-7.32 (2H, m), 4.01 (1H, d, J=9.32 Hz), 3.90 (3H, s), 3.66 (1H,
d, J=9.32 Hz), 3.37 (1H, dd, J=14.86, 1.51 Hz), 2.68-3.08 (5H, m),
2.15-2.25 (1H, m), 2.10-2.14 (1H, m), 1.67-1.77 (1H, m), 1.42-1.63
(2H, m). MS (LC/MS) R.T.=0.81; [M+H].sup.+=340.30.
Example 39
(R)--N-(5-(Difluoromethoxy)pyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00544##
[0365] Step A: 2-Chloro-5-(Difluoromethoxy)pyrimidine
##STR00545##
[0367] 2-Chloropyrimidin-5-ol (1 g, 7.66 mmol) and sodium
2-chloro-2,2-difluoroacetate (3.50 g, 22.98 mmol) in
N,N-dimethylformamide (20 mL) and water (0.2 mL) were heated to
90.degree. C. for 24 hours and concentrated in vacuo. The residue
was purified by column chromatography (5-30% ethyl acetate/hexanes)
to afford 2-chloro-5-(difluoromethoxy)pyrimidine (549 mg, 3.04
mmol, 39.7% yield) as a pale yellow oil. MS (LC/MS) R.T.=1.32;
[M+H].sup.+=181.14.
Step B:
(R)--N-(5-(Difluoromethoxy)pyrimidin-2-yl)-4H-1'-azaspiro[oxazole--
5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00546##
[0369]
(R)--N-(5-(Difluoromethoxy)pyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine was prepared from
2-chloro-5-(difluoromethoxy)pyrimidine by following the general
procedures of Example 23, Steps A-B. .sup.1H NMR (400 MHz,
MeO-d.sub.4) .delta. ppm 8.46 (2H, s), 6.85 (1H, t), 4.02 (1 H, d,
J=10.07 Hz), 3.73 (1H, d, J=10.32 Hz), 3.28 (1H, d, J=1.01 Hz),
3.16 (1H, d), 2.81-3.05 (4H, m), 2.08-2.25 (2H, m), 1.46-1.89 (3H,
m). MS (LC/MS) R.T.=0.53; [M+H].sup.+=326.30.
Example 40
(R)--N-(4,5-Dimethylpyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.-
2.2]octan]-2-amine
##STR00547##
[0370] Step A: 4,5-Dimethylpyrimidin-2-amine
##STR00548##
[0372] A solution of 4-chloro-5,6-dimethylpyrimidin-2-amine (0.35
g, 2.22 mmol) in 2M ammonia in methanol (100 ml) was flushed with
nitrogen and palladium on carbon (0.035 g, 0.33 mmol) was added,
flushed with nitrogen and the reaction was hydrogenated at 1 atm,
ambient temperature for 18 h. The reaction mixture was flushed with
nitrogen and filtered through celite and the celite pad washed with
methanol. The filtrate was evaporated to dryness in vacuo to afford
4,5-dimethylpyrimidin-2-amine (0.35 g, 2.56 mmol, 90% yield) which
was used without further purification. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 7.89 (1H, s), 6.19 (2H, s), 2.18 (3H, s),
1.99 (3H, s). MS (LC/MS) R.T.=0.56; [M+H].sup.+=124.20.
Step B:
(R)--N-(4,5-Dimethylpyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine
##STR00549##
[0374]
(R)--N-(4,5-Dimethylpyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine was prepared from
4,5-dimethylpyrimidin-2-amine by following the general procedures
of Example 23, Steps A-B. .sup.1H NMR (400 MHz, MeOD-d.sub.4)
.delta. ppm 8.22 (1H, s), 4.01 (1H, d, J=10.07 Hz), 3.73 (1H, d,
J=10.32 Hz), 3.40 (1H, d), 3.25 (1H, d), 2.91-3.12 (4H, m), 2.41
(3H, s), 2.09-2.28 (5H, m), 1.62-1.97 (3H, m). MS (LC/MS)
R.T.=0.47; [M+H].sup.+=288.31.
Example 41
(R)--N-(6-Phenylpyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]-
octan]-2-amine
##STR00550##
[0375] Step A: 6-Phenylpyrimidin-4-amine
##STR00551##
[0377] A mixture of 6-chloropyrimidin-4-amine (0.32 g, 2.5 mmol),
phenylboronic acid (0.38 g, 3.13 mmol), saturated aqueous sodium
carbonate (0.80 g, 7.50 mmol) and
bis(triphenylphosphine)palladium(II) chloride (0.035 g, 0.05 mmol)
were suspended in a mixture of dimethoxyethane (15 mL)/ethanol (2
mL)/water (2 mL). The mixture was heated in the microwave at
125.degree. C. for 20 min then concentrated in vacuo. The residue
was purified by column chromatography (10-60% ethyl
acetate/hexanes) to afford 6-phenylpyrimidin-4-amine (167 mg, 0.98
mmol, 39% yield) as an off-white solid. MS (LC/MS) R.T.=0.99;
[M+H].sup.+=172.23.
Step B:
(R)--N-(6-Phenylpyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine
##STR00552##
[0379]
(R)--N-(6-Phenylpyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine was prepared from 6-phenylpyrimidin-4-amine
by following the general procedures of Example 23, Steps A-B.
.sup.1H NMR (400 MHz, MeOD-d.sub.4) .delta. ppm 9.54 (1H, d, J=1.01
Hz), 8.75-8.86 (2H, m), 8.18-8.28 (3H, m), 8.00 (1H, br. s.), 4.70
(1H, d, J=10.32 Hz), 4.43 (1H, d, J=10.58 Hz), 3.72-3.88 (2 H, m),
3.41-3.63 (4H, m), 2.63-2.87 (2H, m), 2.18-2.48 (3H, m). MS (LC/MS)
R.T.=1.36; [M+H].sup.+=336.24.
Example 42
(R)--N-(6-(4-Methoxyphenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00553##
[0381]
(R)--N-(6-(4-Methoxyphenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine was prepared from
6-chloropyrimidin-4-amine by following the general procedures of
Example 41, Steps A-B. .sup.1H NMR (400 MHz, MeOD-d.sub.4) .delta.
ppm 8.72 (1H, d, J=1.26 Hz), 7.90-8.00 (2H, m), 7.16 (1H, br. s.),
6.96-7.05 (2H, m), 4.04 (1H, d, J=10.07 Hz), 3.84 (3H, s), 3.73
(1H, d, J=10.07 Hz), 3.22 (1H, d), 3.09 (1H, d), 2.73-2.98 (4H, m),
2.02-2.21 (2H, m), 1.51-1.85 (3H, m). MS (LC/MS) R.T.=1.44;
[M+H].sup.+=366.28.
Example 43
(R)--N-(6-(6-Methoxypyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine
##STR00554##
[0383]
(R)--N-(6-(6-Methoxypyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxa-
zole-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared from
6-chloropyrimidin-4-amine by following the general procedures of
Example 41, Steps A-B. .sup.1H NMR (400 MHz, MeOD-d.sub.4) .delta.
ppm 8.79 (2H, dd, J=19.01, 1.89 Hz), 8.27 (1H, dd, J=8.56, 2.52
Hz), 7.18 (1H, br. s.), 6.89 (1H, d, J=8.81 Hz), 4.05 (1H, d,
J=10.32 Hz), 3.96 (3H, s), 3.74 (1H, d, J=10.32 Hz), 3.23 (1H, d),
3.10 (1H, d), 2.73-2.99 (4H, m), 2.02-2.21 (2H, m), 1.53-1.84 (3H,
m). MS (LC/MS) R.T.=1.34; [M+H].sup.+=367.25.
Example 44
(R)--N-(6-(Naphthalen-2-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00555##
[0385]
(R)--N-(6-(Naphthalen-2-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine was prepared from
6-chloropyrimidin-4-amine by following the general procedures of
Example 41, Steps A-B. .sup.1H NMR (400 MHz, MeOD-d.sub.4) .delta.
ppm 8.83 (1H, s), 8.56 (1H, s), 7.83-8.13 (4H, m), 7.49-7.58 (2H,
m), 7.37 (1H, br. s.), 4.06 (1H, d, J=10.32 Hz), 3.76 (1H, d,
J=10.32 Hz), 3.23 (1H, s), 3.12 (1H, d), 2.75-3.00 (4H, m),
2.02-2.24 (2H, m), 1.56-1.84 (3H, m). MS (LC/MS) R.T.=1.93;
[M+H].sup.+=386.31.
[0386] The compounds in Table 2 were synthesized according to the
method of Example 1 using the appropriate commercially available
isothiocyanate or amine Amide-containing intermediates were
obtained by the procedures described in Example 3.
TABLE-US-00003 TABLE 2 ##STR00556## Example LCMS RT LCMS Ion Number
R.sub.1 (min) [M + H].sup.+ .sup.1H NMR 45 ##STR00557## 1.48 341.3
.sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 8.60 (1 H, br.
s.), 7.94 (2 H, d, J = 7.63 Hz), 7.43 (1 H, s), 7.39 (2 H, t, J =
7.78 Hz), 7.28 (1 H, t, J = 7.32 Hz), 3.86 (1 H, d, J = 9.77 Hz),
3.60 (1 H, d, J = 9.77 Hz), 2.98- 3.06 (2 H, m), 2.71-2.85 (2 H,
m), 2.66 (2 H, t, J = 7.78 Hz), 2.05 (1 H, br. s.), 1.91 (1 H, br.
s.), 1.53-1.64 (2 H, m), 1.48 (1 H, td, J = 9.99, 7.78 Hz) 46
##STR00558## 1.41 400.4 .sup.1H NMR (500 MHz, DMSO- d.sub.6)
.delta. ppm 8.86 (1 H, br. s.), 7.47 (1 H, d, J = 8.85 Hz), 7.33 (1
H, d, J = 2.44 Hz), 7.01 (1 H, dd, J = 8.85, 2.75 Hz), 3.87 (1 H,
d, J = 10.07 Hz), 3.71- 3.77 (4 H, m), 3.61 (1 H, d, J = 9.77 Hz),
3.06-3.12 (4 H, m), 3.01 (2 H, s), 2.72-2.86 (2 H, m), 2.66 (2 H,
t, J = 7.63 Hz), 2.05 (1 H, br. s.), 1.91 (1 H, br. s.), 1.53-1.64
(2 H, m), 1.43-1.52 (1 H, m) 47 ##STR00559## 1.31 333.3 .sup.1H NMR
(500 MHz, DMSO- d.sub.6) .delta. ppm 9.03 (1 H, br. s.), 7.80 (1 H,
dd, J = 8.70, 5.65 Hz), 7.37 (1 H, dd, J = 10.38, 2.44 Hz), 7.04 (1
H, td, J = 9.08, 2.59 Hz), 3.89 (1 H, d, J = 10.07 Hz), 3.64 (1 H,
d, J = 10.07 Hz), 3.03 (2 H, d, J = 2.75 Hz), 2.74-2.85 (2 H, m),
2.62-2.70 (2 H, m), 2.07 (1 H, d, J = 2.44 Hz), 1.90 (1 H, d, J =
8.85 Hz), 1.54-1.64 (2 H, m), 1.43-1.53 (1 H, m) 48 ##STR00560##
2.07 383.3 .sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 8.00 (1
H, s), 7.41- 7.54 (2 H, m), 3.89 (1 H, d, J = 11.29 Hz), 3.57 (1 H,
d, J = 11.60 Hz), 2.96-3.03 (1 H, m), 2.88-2.93 (1 H, m), 2.73-
2.81 (2 H, m), 2.64 (2 H, dd, J = 8.85, 5.19 Hz), 1.93 (2 H, br.
s.), 1.56 (2 H, br. s.), 1.38- 1.50 (1 H, m) 49 ##STR00561## 2.15
399.4 .sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 9.03 (1 H,
br. s.), 7.91 (1 H, br. s.), 7.65 (1 H, d, J = 8.85 Hz), 7.30 (1 H,
d, J = 7.63 Hz), 3.89 (1 H, d, J = 10.07 Hz), 3.64 (1 H, d, J =
10.07 Hz), 2.97-3.09 (2 H, m), 2.80 (2 H, d, J = 8.55 Hz), 2.66 (2
H, t, J = 7.48 Hz), 2.08 (1 H, br. s.), 1.93 (1 H, br. s.),
1.54-1.66 (2 H, m), 1.44- 1.54 (1 H, m) 50 ##STR00562## 1.89 333.1
.sup.1H NMR (500 MHz, MeOD) .delta. ppm 7.51-7.58 (1 H, m),
7.17-7.25 (1 H, m), 7.08- 7.16 (1 H, m), 4.04-4.13 (1 H, m),
3.72-3.82 (1 H, m), 3.30 (1 H, br. s.), 3.17 (1 H, d, J = 14.95
Hz), 3.01 (2 H, t, J = 7.48 Hz), 2.80-2.92 (2 H, m), 2.12-2.27 (2
H, m), 1.66- 1.88 (3 H, m) 51 ##STR00563## 1.24 333.4 .sup.1H NMR
(500 MHz, DMSO- d.sub.6) .delta. ppm 8.95 (1 H, br. s.), 7.66-7.77
(1 H, m), 7.52- 7.63 (1 H, m), 7.08-7.24 (1 H, m), 3.88 (1 H, d, J
= 9.77 Hz), 3.63 (1 H, d, J = 9.77 Hz), 3.03 (2 H, br. s.), 2.78 (2
H, br. s.), 2.66 (2 H, t, J = 7.17 Hz), 2.06 (1 H, br. s.), 1.91 (1
H, br. s.), 1.39-1.67 (3 H, m) 52 ##STR00564## 2.05 351.2 .sup.1H
NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 9.01 (1 H, br. s.),
7.88-8.00 (1 H, m), 7.53- 7.64 (1 H, m), 3.88 (1 H, d, J = 10.07
Hz), 3.63 (1 H, d, J = 10.07 Hz), 2.98-3.08 (2 H, m), 2.72-2.87 (2
H, m), 2.66 (2 H, t, J = 7.17 Hz), 2.07 (1 H, br. s.), 1.92 (1 H,
br. s.), 1.42- 1.65 (3 H, m) 53 ##STR00565## 1.80 345.3 .sup.1H NMR
(500 MHz, DMSO- d.sub.6) .delta. ppm 9.03 (s, 1 H), 7.65 (d, J =
8.55 Hz, 1 H), 7.19 (d, J = 2.44 Hz, 1 H), 6.82 (dd, J = 8.70, 2.59
Hz, 1 H), 3.90 (d, J = 10.07 Hz, 1 H), 3.79 (s, 3 H), 3.64 (d, J =
10.07 Hz, 1 H), 3.04 (d, J = 2.44 Hz, 2 H), 2.75-2.84 (m, 2 H),
2.67 (t, J = 7.78 Hz, 2 H), 2.07 (s, 1 H), 1.92 (s, 1 H), 1.55-1.63
(m, 2 H), 1.49 (dd, J = 9.77, 2.75 Hz, 1 H) 54 ##STR00566## 1.23
345.2 .sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 8.83 (s, 1
H), 7.37 (d, J = 7.94 Hz, 1 H), 7.14 (t, J = 7.93 Hz, 1 H), 6.92
(d, J = 7.93 Hz, 1 H), 3.89-3.95 (m, 4 H), 3.66 (d, J = 10.07 Hz, 1
H), 3.00-3.08 (m, 2 H), 2.76-2.85 (m, 2 H), 2.67 (t, J = 7.78 Hz, 2
H), 2.08 (s, 1 H), 1.93 (d, J = 3.66 Hz, 1 H), 1.60 (ddd, J =
15.26, 6.87, 3.20 Hz, 2 H), 1.50 (ddd, J = 7.48, 5.19, 2.59 Hz, 1
H) 55 ##STR00567## 1.72 375.2 .sup.1H NMR (500 MHz, DMSO- d.sub.6)
.delta. ppm 8.89 (s, 1 H), 7.40 (s, 1 H), 7.22 (s, 1 H), 3.87 (d, J
= 10.07 Hz, 1 H), 3.79 (d, J = 8.55 Hz, 6 H), 3.62 (d, J = 10.07
Hz, 1 H), 2.99-3.07 (m, 2 H), 2.75-2.84 (m, 2 H), 2.67 (t, J = 7.63
Hz, 2 H), 2.06 (s, 1 H), 1.93 (s, 1 H), 1.56- 1.64 (m, 2 H),
1.46-1.55 (m, 1 H) 56 ##STR00568## 2.14 342.1 .sup.1H NMR (500 MHz,
DMSO- d.sub.6) .delta. ppm 8.75 (s, 1 H), 7.84 (dd, J = 7.93, 1.53
Hz, 3 H), 7.46-7.53 (m, 5 H), 3.85 (d, J = 9.77 Hz, 2 H), 3.60 (d,
J = 10.07 Hz, 2 H), 3.01-3.09 (m, 3 H), 2.84 (t, J = 7.78 Hz, 3 H),
2.67 (t, J = 7.78 Hz, 3 H), 2.09 (s, 2 H), 1.91-1.99 (m, 2 H),
1.53-1.62 (m, 3 H) 57 ##STR00569## 1.04 386.2 .sup.1H NMR (400 MHz,
DMSO- d.sub.6) .delta. ppm 9.05 (1 H, br. s.), 7.86 (1 H, d, J =
1.51 Hz), 7.60 (1 H, d, J = 8.06 Hz), 7.36 (1 H, dd, J = 8.31, 1.76
Hz), 3.90 (1 H, d, J = 10.07 Hz), 3.65 (1 H, d, J = 10.07 Hz), 3.03
(2 H, s), 2.97 (6 H, s), 2.72-2.87 (2 H, m), 2.66 (2 H, t, J = 7.68
Hz), 2.07 (1 H, br. s.), 1.92 (1 H, br. s.), 1.55-1.64 (2 H, m),
1.44-1.54 (1 H, m) 58 ##STR00570## 1.57 426.2 .sup.1H NMR (400 MHz,
DMSO- d.sub.6) .delta. ppm 9.06 (1 H, br. s.), 7.83 (1 H, d, J =
1.51 Hz), 7.61 (1 H, d, J = 8.31 Hz), 7.32 (1 H, dd, J = 8.31, 1.76
Hz), 3.90 (1 H, d, J = 10.07 Hz), 3.65 (1 H, d, J = 10.07 Hz),
3.37-3.57 (4 H, m), 3.04 (2 H, d, J = 1.76 Hz), 2.73-2.86 (2 H, m),
2.66 (2 H, t, J = 7.68 Hz), 2.07 (1 H, d, J = 2.52 Hz), 1.92 (1 H,
dd, J = 8.18, 5.67 Hz), 1.43-1.67 (9 H, m) 59 ##STR00571## 2.17
360.1 .sup.1H NMR (500 MHz, DMSO- D.sub.6) .delta. ppm 8.74 (s, 1
H), 7.86- 7.92 (m, 2 H), 7.31-7.37 (m, 2 H), 3.85 (d, J = 9.77 Hz,
1 H), 3.60 (d, J = 9.77 Hz, 1 H), 3.00-3.09 (m, 2 H), 2.84 (t, J =
7.48 Hz, 2 H), 2.67 (t, J = 7.32 Hz, 2 H), 2.09 (d, J = 1.83 Hz, 1
H), 1.89-1.98 (m, 1 H), 1.55-1.61 (m, 2 H), 1.51-1.54 (m, J = 11.29
Hz, 1 H) 60 ##STR00572## 1.67 332.2 .sup.1H NMR (500 MHz, DMSO-
D.sub.6) .delta. ppm 7.87 (s, 1 H), 7.01 (d, J = 3.05 Hz, 1 H),
6.68 (dd, J = 3.51, 1.68 Hz, 1 H), 3.84 (d, J = 10.38 Hz, 1 H),
3.58 (d, J = 10.07 Hz, 1 H), 3.03 (d, J = 11.29 Hz, 2 H), 2.78-2.86
(m, 2 H), 2.66 (t, J = 7.48 Hz, 2 H), 2.06 (s, 1 H), 1.87-1.96 (m,
1 H), 1.55-1.61 (m, 2 H), 1.51-1.54 (m, 1 H) 61 ##STR00573## 1.83
393.0 .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. ppm 9.02 (1 H,
br. s.), 8.04 (1 H, d, J = 2.01 Hz), 7.49- 7.53 (1 H, m), 7.43-7.47
(1 H, m), 3.88 (1 H, d, J = 10.07 Hz), 3.63 (1 H, d, J = 10.07 Hz),
3.02 (2 H, s), 2.74-2.84 (2 H, m), 2.65 (2 H, t, J = 7.68 Hz), 2.06
(1 H, d, J = 2.52 Hz), 1.89 (1 H, br. s.), 1.42-1.64 (3 H, m) 62
##STR00574## 1.16 391.1 .sup.1H NMR (400 MHz, DMSO- d.sub.6)
.delta. ppm 9.21 (1 H, br. s.), 8.40 (1 H, d, J = 1.51 Hz), 7.80-
7.85 (1 H, m), 7.72-7.77 (1 H, m), 3.92 (1 H, d, J = 10.32 Hz),
3.67 (1 H, d, J = 10.07 Hz), 3.21 (3 H, s), 3.05 (2 H, d, J = 4.03
Hz), 2.82 (2 H, d, J = 6.80 Hz), 2.67 (2 H, t, J = 7.43 Hz), 2.10
(1 H, br. s.), 1.94 (1 H, d, J = 3.27 Hz), 1.45- 1.66 (3 H, m) 63
##STR00575## 1.35 329.1 .sup.1H NMR (500 MHz, DMSO- D.sub.6)
.delta. ppm 8.54 (s, 1 H), 7.32 (s, 1 H), 3.79 (d, J = 10.07 Hz, 1
H), 3.54 (d, J = 10.07 Hz, 1 H), 2.98-3.05 (m, 2 H), 2.76- 2.85 (m,
2 H), 2.66 (t, J = 7.63 Hz, 2 H), 2.05 (s, 1 H), 1.87-1.95 (m, 1
H), 1.41- 1.68 (m, 3 H) 64 ##STR00576## 1.02 349.9 .sup.1H NMR (500
MHz, DMSO- d.sub.6) .delta. ppm 8.54 (s, 1 H), 7.32 (s, 1 H), 3.67
(dd, J = 126.04, 10.07 Hz, 2 H), 2.92-3.09 (m, 2 H), 2.72-2.91 (m,
2 H), 2.60-2.73 (m, J = 7.63, 7.63 Hz, 2 H), 2.00-2.12 (m, 1 H),
1.81-1.99 (m, 1 H), 1.35- 1.69 (m, 3 H) 65 ##STR00577## 2.14 380
.sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 8.93 (s, 1 H),
7.54- 7.76 (m, J = 10.53, 3.81 Hz, 2 H), 3.79-4.04 (m, 1 H), 3.56-
3.72 (m, J = 9.92, 3.20 Hz, 1 H), 3.26-3.45 (m, 2 H), 2.98- 3.12
(m, 2 H), 2.75-2.90 (m, 2 H), 2.61-2.74 (m, J = 1.22 Hz, 2 H), 2.09
(s, 1 H), 1.87-2.04 (m, 1 H), 1.42- 1.76 (m, 3 H) 66 ##STR00578##
1.26 333.1 .sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 9.04 (1
H, d, J = 1.22 Hz), 8.44 (1 H, br. s.), 7.52- 7.69 (2 H, m),
3.77-3.82 (1 H, m), 3.68 (1 H, d, J = 10.07 Hz), 3.05-3.12 (2 H,
m), 2.94 3.04 (2 H, m), 2.80-2.91 (2 H, m), 2.19(1 H, d, J = 1.83
Hz), 2.01 (1 H, br. s.), 1.67- 1.85 (3 H, m) 67 ##STR00579## 0.28
277.1 .sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 6.72-7.61
(m, 1 H), 3.49-3.70 (m, 1 H), 3.31 3.46 (m, 1 H), 2.86-3.06 (m, J =
9.46 Hz, 2 H), 2.56- 2.84 (m, 4 H), 2.10-2.31 (m, 3 H), 1.91-2.10
(m, 4 H), 1.70 (s, 1 H), 1.49-1.61 (m, 2 H), 1.36-1.49 (m, 1 H) 68
##STR00580## 0.44 276.2 .sup.1H NMR (500 MHz, DMSO- d.sub.6)
.delta. ppm 7.55 (s, 1 H), 5.63 (s, 1 H), 3.57-3.74 (m, J = 8.55
Hz, 1 H), 3.48 (s, 3 H), 2.96 (s, 2 H), 2.72-2.88 (m, 2 H), 2.65
(s, 2 H), 1.93-2.17 (m, 4 H), 1.86 (s, 1 H), 1.57 (s, 3 H) 69
##STR00581## 1.35 339.1 .sup.1H NMR (500 MHz, DMSO- d.sub.6)
.delta. ppm 7.87 (d, J = 6.71 Hz, 1 H), 7.80 (d, J = 7.02 Hz, 2 H),
7.42-7.48 (m, 4 H), 3.49- 3.57 (m, 1 H), 3.25-3.33 (m, 4 H), 2.77
(t, J = 15.41 Hz, 2 H), 2.59-2.68 (m, 1 H), 2.49-2.57 (m, 15 H),
2.24- 2.31 (m, 2 H), 2.21 (s, 1 H), 1.94 (s, 1 H), 1.47 (s, 2 H) 70
##STR00582## 0.73 340.1 .sup.1H NMR (300 MHz, DMSO- d.sub.6)
.delta. ppm 9.20 (1 H, br. s.), 8.29-8.42 (1 H, m), 7.63- 7.76 (2
H, m), 3.91 (1 H, d, J = 10.25 Hz), 3.66 (1 H, d, J = 10.25 Hz),
3.05 (2 H, s), 2.74-2.90 (2 H, m), 2.66 (2 H, t, J = 7.68 Hz), 2.09
(1 H, br. s.), 1.92 (1 H, d, J = 4.03 Hz), 1.42-1.66 (3 H, m) 71
##STR00583## 1.00 315.1 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 8.92 (1 H, s), 8.00 (1 H, br. s.), 7.78 (1 H, d, J = 8.56 Hz),
7.42 (1 H, d, J = 8.06 Hz), 3.92 (1 H, d, J = 11.08 Hz), 3.55 (1 H,
d, J = 10.83 Hz), 3.23 (1 H, d, J = 14.86 Hz), 2.83-2.90 (2 H, m),
2.65- 2.83 (2 H, m), 1.89-2.17 (3 H, m), 1.41-1.75 (3 H, m) 72
##STR00584## 0.75 315.1 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 8.82 (1 H, s), 8.22 (1 H, s), 7.98 (1 H, d, J = 8.56 Hz),
7.21-7.25 (1 H, m), 6.37 (1 H, br. s.), 3.99 (1 H, d, J = 11.58
Hz), 3.61 (1 H, d, J = 11.58 Hz), 3.23 (1 H, d, J = 14.86 Hz), 2.89
(2 H, t, J = 7.68 Hz), 2.69-2.84 (2 H, m), 1.91-2.09 (2 H, m),
1.45- 1.73 (4 H, m) 73 ##STR00585## 0.85 309.2 .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. ppm 8.70 (1 H, d, J = 1.76 Hz), 8.09 (1 H,
br. s.), 8.00 (1 H, d, J = 8.31 Hz), 7.43-7.58 (2 H, m), 3.90 (1 H,
d, J = 10.58 Hz), 3.54 (1 H, d, J = 10.32 Hz), 3.26 (1 H, d, J =
14.60 Hz), 2.65-3.06 (5 H, m), 2.13 (1 H, br. s.), 2.00 (1 H, br.
s.), 1.34-1.79 (5 H, m) 74 ##STR00586## 0.65 313.2 .sup.1H NMR (500
MHz, MeOD) .delta. ppm 8.55 (1 H, s), 7.66- 7.70 (1 H, m),
7.59-7.63 (1 H, m), 4.25 (1 H, d, J = 10.99 Hz), 4.04 (1 H, d, J =
10.68 Hz), 3.91 (1 H, d, J = 14.95 Hz), 3.76 (1 H, dd, J = 14.95,
2.44 Hz), 3.45-3.53 (1 H, m), 3.29-3.40 (3 H, m), 2.61 (1 H, d, J =
2.14 Hz), 2.40 (3 H, s), 2.31 (1 H, tt, J = 10.26, 3.47 Hz),
2.06-2.16 (1 H, m, J = 14.23, 9.35, 4.54, 4.54 Hz), 1.92-2.06 (2 H,
m) 75 ##STR00587## 1.24 310.1 .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 8.73 (1 H, d, J = 1.76 Hz), 8.66 (1 H, d, J = 2.01 Hz),
7.96 (1 H, d, J = 9.07 Hz), 7.90 (1 H, br. s.), 7.68 (1 H, br. s.),
7.50 (1 H, br. s.), 3.93 (1 H, d, J = 10.32 Hz), 3.56 (1 H, d, J =
9.32 Hz), 3.29 (1 H, d, J = 14.86 Hz), 2.97 (1 H, d, J = 14.86 Hz),
2.88-2.94 (2 H, m), 2.69-2.87 (2 H, m), 2.12 (1 H, br. s.), 2.06 (1
H, br. s.), 1.64-1.73 (1 H, m), 1.46- 1.64 (2 H, m) 76 ##STR00588##
1.58 328.2 .sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 11.30
(s, 1 H), 9.16 (s, 1 H), 7.04-7.43 (m, 1 H), 6.70-7.03 (m, 1 H),
6.54- 6.69 (m, 1 H), 3.89 (d, J = 9.77 Hz, 1 H), 3.53-3.81 (m, 4
H), 3.01 (s, 2 H), 2.58-2.91 (m, 4 H), 2.03 (s, 1 H), 1.78-1.96 (m,
1 H), 1.32-1.73 (m, 3 H) 77 ##STR00589## 1.40 316.1 .sup.1H NMR
(500 MHz, DMSO- D6) .delta. ppm 11.53 (s, 1 H), 9.12 (s, 1 H),
6.94-7.46 (m, 2 H), 6.83 (s, 2 H), 3.90 (d, J = 9.77 Hz, 1 H), 3.64
(d, J = 9.77 Hz, 1 H), 2.90-3.09 (m, 2 H), 2.61-2.87 (m, 4 H),
1.95-2.10 (m, 1 H), 1.88 (d, J = 3.05 Hz, 1 H), 1.33-1.70 (m, 3 H)
78 ##STR00590## 0.35 298.1 .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 7.94 (1 H, s), 7.80 (1 H, br. s.), 7.26-7.36 (1 H, m),
7.17-7.25 (1 H, m), 3.97 (1 H, d, J = 11.33 Hz), 3.59 (1 H, d, J =
11.58 Hz), 3.23 (1 H, d, J = 14.60 Hz), 2.65-3.01 (5 H, m),
1.86-2.16 (2 H, m), 1.35- 1.73 (3 H, m) 79 ##STR00591## 1.10 376.1
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.78 (2 H, d, J =
5.79 Hz), 8.03 (2 H, d, J = 6.04 Hz), 7.73 (1 H, br. s.), 7.48 (1
H, d, J = 8.56 Hz), 7.36 (1 H, br. s.), 3.92 (1 H, d, J = 8.81 Hz),
3.55 (1 H, d, J = 9.06 Hz), 3.24 (1 H, d, J = 14.10 Hz), 2.57-3.06
(5 H, m), 1.86-2.20 (2 H, m), 1.30-1.80 (3 H, m) 80 ##STR00592##
2.36 393.1 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.00 (1 H,
d, J = 7.81 Hz), 7.83-7.97 (1 H, m), 7.68 (1 H, br. s.), 7.39-7.55
(2 H, m), 7.35 (1 H, br. s.), 7.20 (2 H, td, J = 8.31, 2.27 Hz),
3.93 (1 H, d, J = 10.83 Hz), 3.56 (1 H, d, J = 10.58 Hz), 3.24 (1
H, d, J = 15.11 Hz), 2.62-3.05 (5 H, m), 1.88-2.22 (2 H, m), 1.34-
1.79 (3 H, m) 81 ##STR00593## 2.81 403.1 .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 8.12 (2 H, d, J = 8.06 Hz), 7.64 (1 H, br.
s.), 7.43 (1 H, d, J = 8.56 Hz), 7.32 (3 H, d, J = 8.06 Hz), 3.94
(1 H, d, J = 10.07 Hz), 3.56 (1 H, d, J = 9.57 Hz), 3.24 (1 H, d, J
= 13.60 Hz), 2.57-3.03 (7 H, m), 1.88-2.22 (2 H, m), 1.39- 1.74 (3
H, m), 1.27 (3 H, t, J = 7.68 Hz) 82 ##STR00594## 2.62 418.1
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.06 (2 H, d, J =
9.07 Hz), 7.56 (1 H, br. s.), 7.30-7.43 (2 H, m), 6.74 (2 H, d, J =
9.07 Hz), 3.95 (1 H, d, J = 11.58 Hz), 3.58 (1 H, d, J = 11.83 Hz),
3.24 (1 H, d, J = 14.10 Hz), 3.05 (6 H, s), 2.61-2.99 (5 H, m),
1.88-2.13 (2 H, m), 1.35-1.81 (4 H, m) 83 ##STR00595## 1.15 367.1
.sup.1H NMR (500 MHz, MeOD) .delta. ppm 9.12 (1 H, s), 7.78- 7.82
(1 H, m), 7.70-7.74 (1 H, m), 4.05 (1 H, d, J = 9.77 Hz), 3.75 (1
H, d, J = 9.77 Hz), 3.29 (1 H, d, J = 14.65 Hz), 3.11-3.18 (1 H,
m), 2.98 (2 H, t, J = 7.93 Hz), 2.80-2.91 (2 H, m), 2.13-2.22 (2
H,
m), 1.71-1.87 (2 H, m), 1.62- 1.71 (1 H, m) 84 ##STR00596## 1.96
393.1 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.18-8.26 (2 H,
m), 7.66 (1 H, s), 7.44 (1 H, d, J = 8.56 Hz), 7.32 (1 H, d, J =
8.56 Hz), 7.16-7.23 (2 H, m), 3.95 (1 H, d, J = 11.33 Hz), 3.57 (1
H, d, J = 11.33 Hz), 3.25 (1 H, d, J = 14.60 Hz), 2.69-3.04 (6 H,
m), 2.09 (1 H, br. s.), 1.45- 1.74 (4 H, m) 85 ##STR00597## 1.34
309.1 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.75 (1 H, dd,
J = 4.15, 1.64 Hz), 8.04 (1 H, d, J = 8.06 Hz), 7.98 (2 H, d, J =
8.81 Hz), 7.49 (1 H, d, J = 8.81 Hz), 7.31 (1 H, dd, J = 8.31, 4.28
Hz), 6.71 (1 H, br. s.), 4.03 (1 H, d, J = 11.83 Hz), 3.65 (1 H, d,
J = 12.09 Hz), 3.25 (1 H, d, J = 14.86 Hz), 2.96 (1 H, d, J = 14.86
Hz), 2.89 (2 H, t, J = 7.55 Hz), 2.69-2.84 (2 H, m), 2.07 (1 H, br.
s.), 1.98 (1 H, br. s.), 1.57-1.73 (2 H, m), 1.44-1.55 (1 H, m) 86
##STR00598## 1.16 313.1 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 9.78 (1 H, br. s.), 7.19 (1 H, br. s.), 6.99 (1 H, d, J = 6.80
Hz), 6.67 (1 H, d, J = 8.31 Hz), 3.87 (1 H, d, J = 10.32 Hz), 3.50
(1 H, d, J = 11.08 Hz), 3.46 (2 H, s), 3.21 (1 H, d, J = 14.60 Hz),
2.93 (1 H, d, J = 15.11 Hz), 2.87 (2 H, t, J = 7.68 Hz), 2.65- 2.81
(2 H, m), 2.04 (1 H, br. s.), 1.96 (1 H, br. s.), 1.40- 1.75 (3 H,
m) 87 ##STR00599## 1.13 299.2 .sup.1H NMR (500 MHz, MeOD) .delta.
ppm 8.57-8.63 (1 H, m), 7.55-7.64 (2 H, m), 7.06- 7.12 (1 H, m),
4.04 (1 H, dd, J = 9.92, 1.98 Hz), 3.81 (1 H, dd, J = 10.07, 2.14
Hz), 3.46- 3.57 (1 H, m), 3.23 (2 H, d, J = 10.68 Hz), 3.09-3.18 (2
H, m), 2.82-3.02 (1 H, m), 2.41 (1 H, br. s.), 2.28-2.37 (1 H, m),
1.99-2.07 (1 H, m), 1.86- 1.97 (2 H, m) 88 ##STR00600## 1.26 342.1
.sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 9.20 (s, 1 H),
8.64 (s, 1 H), 8.45-8.56 (m, J = 4.58, 1.53 Hz, 1 H), 8.21- 8.38
(m, J = 7.63 Hz, 1 H), 7.62 (s, 1 H), 7.33-7.50 (m, J = 7.93, 4.88
Hz, 1 H), 3.75 (dd, J = 128.64, 9.92 Hz, 2 H), 2.95-3.14 (m, 2 H),
2.62- 2.91 (m, 4 H), 2.06 (s, 1 H), 1.92 (s, 1 H), 1.42-1.70 (m, 3
H) 89 ##STR00601## 2.13 343.1 .sup.1H NMR (500 MHz, DMSO- d.sub.6)
.delta. ppm 8.81 (s, 1 H), 7.39 (s, 1 H), 6.97 (s, 1 H), 3.78 (dd,
J = 130.16, 9.92 Hz, 2 H), 2.95-3.15 (m, 2 H), 2.74- 2.90 (m, 2 H),
2.61-2.72 (m, J = 7.78, 7.78 Hz, 2 H), 2.47- 2.55 (m, J = 3.66,
1.83 Hz, 3 H), 2.33 (s, 3 H), 2.07 (s, 1 H), 1.85-2.01 (m, 1 H),
1.41- 1.70 (m, 3 H) 90 ##STR00602## 0.55 372.0 .sup.1H NMR (300
MHz, DMSO- d.sub.6) .delta. ppm 9.98 (1 H, s), 8.92 (1 H, br. s.),
8.11 (1 H, d, J = 1.83 Hz), 7.48-7.53 (1 H, m), 7.34-7.40 (1 H, m),
3.88 (1 H, d, J = 10.25 Hz), 3.62 (1 H, d, J = 10.25 Hz), 3.02 (2
H, s), 2.74-2.83 (2 H, m), 2.66 (2 H, t, J = 7.50 Hz), 2.04 (4 H,
s), 1.90 (1 H, br. s.), 1.58 (3 H, br. s.) 91 ##STR00603## 1.60
343.3 .sup.1H NMR (300 MHz, DMSO- d.sub.6) .delta. ppm 7.61 (1 H,
s), 7.47- 7.53 (1 H, m), 7.16(1 H, d, J = 8.05 Hz), 3.84-3.92 (1 H,
m), 3.62 (1 H, d, J = 10.25 Hz), 3.26-3.46 (3 H, m), 3.02 (1 H, s),
2.79 (1 H, d, J = 6.95 Hz), 2.65 (2 H, q, J = 7.32 Hz), 2.01-2.20
(1 H, m), 1.93 (2 H, d, J = 9.88 Hz), 1.40-1.64 (2 H, m), 1.15-1.28
(3 H, m), 1.00 (1 H, d, J = 6.59 Hz) 92 ##STR00604## 0.45 279.1
.sup.1H NMR (500 MHz, DMSO- D6) .delta. ppm 8.34-8.83 (m, 1 H),
6.96 (s, 1 H), 3.79 (d, J = 9.77 Hz, 2 H), 3.53 (d, J = 9.77 Hz, 2
H), 2.89-3.06 (m, 2 H), 2.71-2.86 (m, 2 H), 2.60-2.70 (m, J = 7.78,
7.78 Hz, 2 H), 2.29 (s, 3 H), 2.01 (s, 1 H), 1.90 (d, J = 13.43 Hz,
2 H), 1.35-1.65 (m, 3 H) 93 ##STR00605## 1.79 355.1 .sup.1H NMR
(500 MHz, DMSO- d.sub.6) .delta. ppm 8.74 (s, 1 H),7.62- 7.88 (m, 1
H), 3.83 (d, J = 10.07 Hz, 1 H), 3.63-3.76 (m, 2 H), 3.58 (d, J =
10.07 Hz, 1 H), 3.40-3.53 (m, 2 H), 2.96-3.10 (m, 2 H), 2.73- 2.90
(m, 2 H), 2.58-2.72 (m, J = 7.63, 7.63 Hz, 2 H), 2.06 (s, 1 H),
1.75-1.99 (m, 5 H), 1.43-1.69 (m, 3 H) 94 ##STR00606## 1.53 313.1
.sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.63 (1 H, d, J = 7.02 Hz),
7.55 (1 H, s), 7.19-7.23 (1 H, m), 4.27 (1 H, d, J = 10.68 Hz),
4.06 (1 H, d, J = 10.68 Hz), 3.94 (1 H, d, J = 14.95 Hz), 3.76-3.83
(1 H, m), 3.54 (1 H, t, J = 11.90 Hz), 3.36-3.48 (3 H, m), 2.64 (1
H, d, J = 1.83 Hz), 2.56 (3 H, s), 2.34-2.42 (1 H, m, J = 10.15,
10.15, 3.66, 3.51 Hz), 2.17 (1 H, dddd, J = 14.19, 9.46, 4.58, 4.43
Hz), 1.97-2.11 (2 H, m) 95 ##STR00607## 0.63 260.2 .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. ppm 9.05 (1 H, br. s.), 8.34 (1 H, s),
8.04 (1 H, dd, J = 2.77, 1.26 Hz), 8.00 (1 H, d, J = 2.77 Hz), 3.91
(1 H, d, J = 9.32 Hz), 3.57 (1 H, d, J = 9.57 Hz), 3.31 (1 H, dd, J
= 14.98, 1.64 Hz), 2.65-3.02 (5 H, m), 2.10-2.20 (1 H, m),
2.05-2.09 (1 H, m), 1.63- 1.75 (1 H, m), 1.40-1.60 (2 H, m) 96
##STR00608## 1.88 338.1 .sup.1H NMR (500 MHz, DMSO- D6) .delta. ppm
8.85 (s, 1 H), 7.41 7.47 (m, 4 H), 7.28-7.35 (m, 1 H), 6.85 (s, 1
H), 3.83 (d, J = 9.77 Hz, 1 H), 3.57 (d, J = 9.77 Hz, 1 H),
3.46-3.51 (m, 3 H), 3.00 (s, 2 H), 2.78 (t, J = 7.78 Hz, 2 H), 2.67
(t, J = 7.78 Hz, 2 H), 1.98-2.03 (m, 1 H), 1.85-1.93 (m, 1 H),
1.54-1.63 (m, J = 8.16, 7.82, 7.82, 3.05 Hz, 2 H), 1.43- 1.51 (m, 1
H) 97 ##STR00609## 1.57 313.1 .sup.1H NMR (500 MHz, MeOD) .delta.
ppm 8.61 (1 H, d, J = 6.71 Hz), 7.65 (1 H, d, J = 7.32 Hz), 7.25 (1
H, t, J = 7.02 Hz), 4.32 (1 H, d, J = 10.99 Hz), 4.11 (1 H, d, J =
10.68 Hz), 3.98 (1 H, dd, J = 14.80, 1.68 Hz), 3.82 (1 H, dd, J =
14.95, 2.14 Hz), 3.52- 3.61 (1 H, m), 3.35-3.47 (3 H, m), 2.66-2.71
(1 H, m), 2.62 (3 H, s), 2.34-2.44 (1 H, m), 2.14-2.24 (1 H, m),
1.99- 2.13 (2 H, m) 98 ##STR00610## 0.57 350.1 .sup.1H NMR (500
MHz, DMSO- D6) .delta. ppm 8.64 (s, 1 H), 3.82 (d, J = 10.07 Hz, 1
H), 3.57 (d, J = 9.77 Hz, 1 H), 3.02 (s, 2 H), 2.97 (s, 6 H), 2.79
(s, 2 H), 2.67 (t, J = 7.63 Hz, 2 H), 2.19 (s, 3 H), 2.04 (s, 1 H),
1.85- 1.94 (m, 1 H), 1.58 (s, 2 H), 1.49 (d, J = 6.71 Hz, 1 H) 99
##STR00611## 0.87 290.1 .sup.1H NMR (400 MHz, MeOD) .delta. ppm
8.20 (1 H, d, J = 5.79 Hz), 6.34 (1 H, d, J = 5.79 Hz), 3.94-4.04
(1 H, m), 3.91 (3 H, s), 3.55-3.76 (1 H, m), 3.00-3.28 (2 H, m),
2.63- 2.97 (4 H, m), 1.99-2.21 (2 H, m), 1.44-1.85 (3 H, m) 100
##STR00612## 0.91 290.1 .sup.1H NMR (400 MHz, MeOD) .delta. ppm
8.39 (1 H, s), 6.21 (1 H, d, J = 6.04 Hz), 3.98 (1 H, d, J = 10.32
Hz), 3.89 (3 H, s), 3.67 (1 H, d, J = 10.32 Hz), 3.01-3.24 (2 H,
m), 2.65- 2.97 (4 H, m), 1.96-2.17 (2 H, m), 1.43-1.81 (3 H, m) 101
##STR00613## 1.18 260.1 .sup.1H NMR (400 MHz, MeOD) .delta. ppm
8.56 (2 H, d, J = 4.78 Hz), 6.94-6.99 (1 H, m), 3.98- 4.04 (1 H,
m), 3.78 (1 H, d, J = 10.32 Hz), 3.45-3.52 (1 H, m), 3.35 (1 H, s),
3.15-3.22 (2 H, m), 3.05-3.12 (2 H, m), 2.22-2.38 (2 H, m), 1.95-
2.03 (1 H, m), 1.80-1.92 (2 H, m) 102 ##STR00614## 2.19 343.4
.sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 8.87 (1 H, br.
s.), 7.06 (1 H, d, J = 7.63 Hz), 6.91 (1 H, d, J = 7.63 Hz), 3.92
(1 H, d, J = 10.07 Hz), 3.66 (1 H, d, J = 10.07 Hz), 3.03 (2 H, s),
2.72-2.85 (2 H, m), 2.64- 2.70 (2 H, m), 2.51 (3 H, s), 2.35 (3 H,
s), 2.07 (1 H, br. s.), 1.91 (1 H, br. s.), 1.55- 1.65 (2 H, m),
1.44-1.53 (1 H, m) 103 ##STR00615## 2.48 363.3 .sup.1H NMR (500
MHz, DMSO- d.sub.6) .delta. ppm 8.84 (1 H, br. s.), 7.71 (1 H, d, J
= 1.83 Hz), 7.18 (1 H, s), 3.90 (1 H, d, J = 10.07 Hz), 3.64 (1 H,
d, J = 10.07 Hz), 3.03 (2 H, s), 2.72-2.85 (2 H, m), 2.66 (2 H, t,
J = 7.63 Hz), 2.53 (3 H, s), 2.07 (1 H, br. s.), 1.88-1.96 (1 H,
m), 1.55-1.63 (2 H, m), 1.44- 1.53 (1 H, m) 104 ##STR00616## 1.45
381.0 .sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 8.99 (1 H,
br. s.), 7.69 (1 H, d, J = 2.44 Hz), 7.61 (1 H, d,J = 8.55Hz),7.01-
7.34 (2 H, m), 3.89 (1 H, d, J = 10.07 Hz), 3.64 (1 H, d, J = 10.07
Hz), 3.03 (2 H, d, J = 2.44 Hz), 2.72-2.86 (2 H, m), 2.66 (2 H, t,
J = 7.78 Hz), 2.07 (1 H, br. s.), 1.88-1.96 (1 H, m), 1.55-1.64 (2
H, m), 1.45-1.53 (1 H, m) 105 ##STR00617## 1.83 355.1 .sup.1H NMR
(500 MHz, DMS0- D6) .delta. ppm 8.57 (s, 1 H), 7.31 (t, J = 7.32
Hz, 2 H), 7.20- 7.26 (m, 3 H), 7.08 (s, 1 H), 4.01 (s, 2 H), 3.78
(d, J = 9.77 Hz, 1 H), 3.53 (d, J = 10.07 Hz, 1 H), 2.97 (s, 2 H),
2.70-2.78 (m, 2 H), 2.64 (t, J = 7.78 Hz, 2 H), 1.99 (s, 1 H),
1.81-1.90 (m, 1 H), 1.52-1.60 (m, 2 H), 1.41-1.49 (m, J = 6.90,
2.90, 2.67, 2.48 Hz, 1 H) 106 ##STR00618## 0.71 274.2 .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. ppm 8.87 (1 H, br. s.), 8.23 (1 H,
s), 7.88 (1 H, s), 3.85 (1 H, d, J = 9.32 Hz), 3.51 (1 H, d, J =
9.32 Hz), 3.27 (1 H, dd, J = 14.98, 1.64 Hz), 2.60-2.97 (5 H, m),
2.37 (3 H, s), 2.06- 2.16 (1 H, m, J = 13.25, 9.85, 3.46, 3.46,
3.46 Hz), 1.99- 2.05 (1 H, m), 1.57-1.70 (1 H, m), 1.36-1.54 (2 H,
m) 107 ##STR00619## 0.93 338.1 .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 8.71 (1 H, br. s.), 8.14 (1 H, d, J = 1.26 Hz), 8.11 (1
H, d, J = 1.26 Hz), 3.92 (1 H, d, J = 9.32 Hz), 3.58 (1 H, d, J =
9.57 Hz), 3.32 (1 H, dd, J = 14.98, 1.89 Hz), 2.68-3.03 (5 H, m),
2.04-2.19 (2 H, m), 1.63-1.75 (1 H, m, J = 14.01, 9.85, 4.31, 4.31
Hz), 1.42- 1.59 (2 H, m) 108 ##STR00620## 2.34 375.0 .sup.1H NMR
(500 MHz, DMSO- D6) .delta. ppm 8.58 (s, 1 H), 7.98 (d, J = 8.24
Hz, 2 H), 7.51 (s, 1 H), 7.45 (d, J = 8.54 Hz, 2 H), 3.87 (d, J =
10.07 Hz, 1 H), 3.62 (d, J = 9.77 Hz, 1 H), 3.04 (s, 2 H),
2.75-2.87 (m, 2 H), 2.68 (t, J = 7.63 Hz, 2 H), 2.07 (s, 1 H), 1.92
(s, 1 H), 1.40- 1.68 (m, 3 H) 109 ##STR00621## 0.73 279.3 .sup.1H
NMR (400 MHz, DMSO- D6) .delta. ppm 8.61 (s, 1 H), 6.54 (d, J =
1.01 Hz, 1 H), 3.80 (d, J = 10.07 Hz, 1 H), 3.54 (d, J = 9.82 Hz, 1
H), 2.98 (s, 2 H), 2.70-2.81 (m, 2 H), 2.65 (t, J = 7.81 Hz, 2 H),
2.20 (s, 3 H), 1.95-2.05 (m, 1 H), 1.88 (s, 1 H), 1.50-1.61 (m, 2
H), 1.41- 1.50 (m, 1 H) 110 ##STR00622## 1.76 321.4 .sup.1H NMR
(400 MHz, DMSO- D6) .delta. ppm 8.24-8.82 (m, 1 H), 6.51 (s, 1 H),
3.83 (d, J = 9.57 Hz, 1 H), 3.57 (d, J = 9.32 Hz, 1 H), 2.98 (s, 2
H), 2.70-2.82 (m, 2 H), 2.65 (t, J = 7.68 Hz, 2 H), 1.97-2.04 (m, 1
H), 1.88 (s, 1 H), 1.52- 1.61 (m, 2 H), 1.46 (dd, J = 9.69, 2.90
Hz, 1 H), 1.24 (s, 9 H) 111 ##STR00623## 1.64 312.3 .sup.1H NMR
(400 MHz, DMSO- d.sub.6) .delta. ppm 8.55 (s, 1 H), 7.13 (d, J =
8.56 Hz, 2 H), 7.04 (s, 1 H), 6.86 (d, J = 8.56 Hz, 2 H), 3.92 (s,
2 H), 3.66-3.85 (m, 4 H), 3.45-3.60 (m, J = 10.07 Hz, 1 H), 2.95
(s, 2 H), 2.57- 2.86 (m, 4 H), 1.98 (s, 1 H), 1.84 (s, 1 H),
1.31-1.66 (m, 2 H) 112 ##STR00624## 1.70 359.1 .sup.1H NMR (300
MHz, DMSO- d.sub.6) .delta. ppm 8.83 (1 H, d, J = 1.83 Hz), 7.08 (1
H, d, J = 8.42 Hz), 6.71 (1 H, d, J = 8.05 Hz), 3.87- 3.94 (1 H,
m), 3.85 (3 H, s), 3.65 (1 H, d, J = 9.88 Hz), 3.04 (2 H, s),
2.61-2.85 (4 H, m), 2.47 (3 H, s), 2.08 (1 H, d, J = 2.20 Hz),
1.82-1.99 (1 H, m), 1.41-1.66 (3 H, m) 113 ##STR00625## 1.48 298.3
.sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 12.09 (s, 1 H),
8.02 (s, 1 H), 7.63 (d, J = 7.32 Hz, 1 H), 7.33-7.36 (m, 1 H), 7.30
(d, J = 5.80 Hz, 1 H), 6.99 (t, J = 7.02 Hz, 1 H), 3.82 (d, J =
8.24 Hz, 1 H), 3.57 (s, 1 H), 3.00 (s, 2 H), 2.79 (s, 2 H), 2.67
(s, 2 H), 1.99-2.04 (m, 1 H), 1.93 (s, 1 H), 1.58 (s, 2 H), 1.47
(s, 1 H)
[0387] The compounds in Table 3 were synthesized according to the
method of Example 21, steps C-D using the appropriate
isothiocyanate or amine and racemic
3-(aminomethyl)quinuclidin-3-ol, 2 HCl.
TABLE-US-00004 TABLE 3 ##STR00626## Example LCMS RT LCMS Ion Number
R.sub.1 (min) [M + H].sup.+ .sup.1H NMR 114 ##STR00627## 2.06 344.3
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. ppm 9.02 (1 H, s),
7.10 (1 H, s), 3.67-3.88 (2 H, m), 3.07-3.30 (2 H, m), 2.80- 3.05
(4 H, m), 2.78 (3 H, s), 2.55 (3 H, s), 2.22 (1 H, br. s.), 2.00 (1
H, d, J = 12.09 Hz), 1.55-1.88 (3 H, m) 115 ##STR00628## 1.19 260.3
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.70 (1 H, dd, J = 4.41,
1.39 Hz), 7.51 (1 H, dd, J = 8.81, 4.53 Hz), 7.15 (1 H, d, J = 7.81
Hz), 4.03 (1 H, d, J = 10.07 Hz), 3.72 (1 H, d, J = 10.07 Hz),
3.14-3.26 (1 H, m), 3.03-3.14 (1 H, m), 2.85- 2.99 (2 H, m),
2.69-2.84 (2 H, m), 2.00-2.26 (2 H, m), 1.51-1.87 (3 H, m) 116
##STR00629## 0.73 290.3 .sup.1H NMR (400 MHz, MeOD) .delta. ppm
6.99-7.12 (2 H, m), 3.95-4.02 (4 H, m), 3.68 (1 H, d, J = 10.07
Hz), 3.20 (1 H, dd), 3.08 (1 H, dd), 2.84- 2.95 (2 H, m), 2.69-2.84
(2 H, m), 1.99-2.21 (2 H, m), 1.47-1.87 (3 H, m) 117 ##STR00630##
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.71 (1 H, s), 8.33 (1 H,
d, J = 5.54 Hz), 6.84 (1 H, br. s.), 4.04 (1 H, d, J = 10.32 Hz),
3.73 (1 H, d, J = 10.32 Hz), 3.17-3.25 (1 H, m), 3.04- 3.13 (1 H,
m), 2.70-3.00 (4 H, m), 2.02-2.18 (2 H, m), 1.51-1.81 (3 H, m) 118
##STR00631## 0.83 290.3 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 8.48 (1 H, br. s.), 7.96 (1 H, br. s.), 7.71 (1 H, s), 3.91 (1
H, d, J = 9.32 Hz), 3.87 (3 H, s), 3.57 (1 H, d, J = 9.32 Hz), 3.30
(1 H, d, J = 14.86 Hz), 2.62-3.03 (5 H, m), 2.09- 2.21 (1 H, m),
2.08 (1 H, br. s.), 1.61-1.78 (1 H, m), 1.39- 1.60 (2 H, m) 119
##STR00632## 0.84 290.3 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 8.56 (1 H, br. s.), 7.96 (1 H, d, J = 1.26 Hz), 7.79 (1 H, d, J
= 1.51 Hz), 3.89 (3 H, s), 3.85 (1 H, s), 3.53 (1 H, d, J = 9.06
Hz), 3.31 (1 H, d, J = 14.86 Hz), 2.65-3.04 (5 H, m), 2.11-2.23 (1
H, m, J = 9.85, 9.85, 6.74, 3.53 Hz), 2.08 (1 H, br. s.), 1.62-1.75
(1 H, m), 1.40-1.61 (2 H, m) 120 ##STR00633## 0.75 290.3 .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.30 (1 H, br. s.), 7.58 (1
H, d, J = 3.02 Hz), 7.52 (1 H, d, J = 3.02 Hz), 3.92 (3 H, s), 3.88
(1 H, d, J = 9.32 Hz), 3.54 (1 H, d, J = 9.32 Hz), 3.28 (1 H, dd, J
= 14.86, 1.51 Hz), 2.63-2.96 (5 H, m), 2.07- 2.22 (1 H, m), 2.02 (1
H, br. s.), 1.57-1.71 (1 H, m), 1.33- 1.55 (2 H, m) 121
##STR00634## 1.15 316.4 .sup.1H NMR (400 MHz, MeOD) .delta. ppm
8.65 (1 H, d, J = 3.02 Hz), 6.82 (1 H, br. s.), 4.02 (1 H, dd, J =
10.32, 3.02 Hz), 3.71 (1 H, dd, J = 10.32, 3.02 Hz), 2.98-3.26 (2
H, m), 2.66- 2.96 (4 H, m), 1.97-2.18 (2 H, m), 1.48-1.86 (3 H, m),
1.29 (9 H, s) 122 ##STR00635## 1.73 309.3 .sup.1H NMR (400 MHz,
MeOD) .delta. ppm 8.99 (1 H, s), 7.90 (1 H, d, J = 8.31 Hz), 7.70
(1 H, d, J = 8.31 Hz), 7.57 (1 H, t, J = 7.55 Hz), 7.39 (1 H, t, J
= 7.55 Hz), 7.30 (1 H, br. s.), 3.94 (1 H, d, J = 9.82 Hz), 3.63 (1
H, d, J = 10.07 Hz), 3.14- 3.24 (1 H, m), 3.00-3.10 (1 H, m),
2.68-2.98 (4 H, m), 2.01-2.24 (2 H, m), 1.45- 1.87 (3 H, m) 123
##STR00636## 0.79 288.30 .sup.1H NMR (400 MHz, MeOD- d.sub.4)
.delta. ppm 6.51 (1 H, br. s.), 4.03 (1 H, d, J = 10.32 Hz), 3.71
(1 H, d, J = 10.32 Hz), 3.12-3.24 (1 H, m), 3.01-. 3.12 (1 H, m),
2.72-3.00 (4 H, m), 2.52 (3 H, s), 2.33 (3 H, s), 1.98-2.20 (2 H,
m), 1.51-1.87 (3 H, m) 124 ##STR00637## 0.72 274.30 .sup.1H NMR
(400 MHz, MeOD- d.sub.4) .delta. ppm 7.42 (1 H, d, J = 9.07 Hz),
7.07 (1 H, d, J = 8.06 Hz), 4.02 (1 H, d, J = 10.07 Hz), 3.71 (1 H,
d, J = 10.07 Hz), 3.15-3.25 (1 H, m), 3.04- 3.14 (1 H, m),
2.69-2.98 (4 H, m), 2.55 (3 H, s), 1.99- 2.21 (2 H, m), 1.47-1.89
(3 H, m)
[0388] The compounds in Table 3 were synthesized according to the
method of Example 21, steps C-D using the appropriate
isothiocyanate or amine and (S)-3-(aminomethyl)quinuclidin-3-ol, 2
HCl.
TABLE-US-00005 TABLE 3 ##STR00638## LCMS Example LCMS Ion Number
R.sub.1 RT (min) [M + H].sup.+ .sup.1H NMR 125 ##STR00639## 2.06
329.11 .sup.1H NMR (500 MHz, MeOD- d.sub.4) .delta. ppm 8.37 (d, J
= 7.32 Hz, 1 H), 6.92 (d, J = 2.75 Hz, 1 H), 6.65-6.75 (m, 1 H),
4.01 (d, J = 9.77 Hz, 1 H), 3.86-3.95 (m, 3 H), 3.70 (d, J = 9.77
Hz, 1 H), 3.26 (d, J = 14.95 Hz, 2 H), 3.12 (d, J = 14.95 Hz, 1 H),
2.96 (t, J = 7.32 Hz, 2 H), 2.76- 2.89 (m, 2 H), 2.06-2.26 (m, 2
H), 1.58-1.86 (m, 3 H) 126 ##STR00640## 0.56 260.30 .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. ppm 8.52 (2 H, d, J = 4.78 Hz), 6.83
(1 H, t, J = 4.78 Hz), 3.94- 4.10 (2 H, m), 3.87 (1 H, d, J = 14.60
Hz), 3.62 (1 H, dd, J = 14.48, 1.89 Hz), 3.41-3.56 (1 H, m), 3.29
(2 H, t, J = 8.44 Hz), 3.04-3.18 (1 H, m), 2.27- 2.50 (2 H, m),
1.88-2.06 (2 H, m), 1.71-1.87 (1 H, m) 127 ##STR00641## 0.88 294.20
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.58 (1 H, br. s.),
8.18 (1 H, s), 7.97 (1 H, s), 3.93 (1 H, d, J = 9.32 Hz), 3.58 (1
H, d, J = 9.57 Hz), 3.28 (1 H, dd, J = 14.86, 1.51 Hz), 2.64-2.98
(5 H, m), 2.02-2.15 (2 H, m), 1.60-1.73 (1 H, m), 1.38- 1.56 (2 H,
m) 128 ##STR00642## 0.86 294.20 .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 8.72 (1 H, br. s.), 8.12 (1 H, d, J = 0.76 Hz), 8.06 (1
H, d, J = 1.26 Hz), 3.93 (1 H, d, J = 9.32 Hz), 3.59 (1 H, d, J =
9.32 Hz), 3.32 (1 H, dd, J = 14.98, 1.38 Hz), 2.68-3.02 (5 H, m),
2.05-2.21 (2 H, m), 1.64-1.76 (1 H, m), 1.42- 1.59 (2 H, m). M.P..
185-8.degree. C. 129 ##STR00643## 2.04 385.28 .sup.1H NMR (500 MHz,
DMSO- D.sub.6) .delta. ppm 8.48 (s, 1 H), 7.58 (d, J = 8.24 Hz, 2
H), 6.98 (d, J = 8.55 Hz, 2 H), 3.73-3.87 (m, 4 H), 3.56 (d, J =
9.16 Hz, 1 H), 3.00 (s, 2 H), 2.72-2.87 (m, 2 H), 2.61-2.72 (m, J =
7.63, 7.63 Hz, 2 H), 2.38 (s, 3 H), 2.03 (s, 1 H), 1.91 (s, 1 H),
1.37-1.70 (m, 3 H) 130 ##STR00644## 1.10 368.20 .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. ppm 8.97 (1 H, br. s.), 7.67 (1 H, s),
3.94 (3 H, s), 3.89 (1 H, d, J = 9.57 Hz), 3.55 (1 H, d, J = 9.57
Hz), 3.28 (1 H, dd, J = 14.86, 1.76 Hz), 2.63-2.95 (5 H, m),
2.08-2.19 (1 H, m), 1.99-2.06 (1 H, m), 1.60- 1.70 (1 H, m, J =
14.01, 9.85, 4.31, 4.31 Hz), 1.35-1.55 (2 H, m) 131 ##STR00645##
1.19 336.30 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.06 (1
H, br. s.), 8.52 (1 H, d, J = 1.51 Hz), 8.44 (1 H, d, J = 1.26 Hz),
7.89-7.93 (2 H, m), 7.40-7.47 (2 H, m), 7.33- 7.39 (1 H, m), 3.94
(1 H, d, J = 9.32 Hz), 3.60 (1 H, d, J = 9.32 Hz), 3.34 (1 H, dd, J
= 14.86, 1.76 Hz), 2.69-3.05 (5 H, m), 2.13-2.23 (1 H, m),
2.07-2.12 (1 H, m), 1.65- 1.76 (1 H, m), 1.44-1.60 (2 H, m) 132
##STR00646## 1.35 332.23 .sup.1H NMR (500 MHz, DMSO- D6) .delta.
ppm 12.40 (s, 1 H), 7.95 (s, 1 H), 7.30 (d, J = 8.24 Hz, 1 H), 7.22
(t, J = 7.78 Hz, 2 H), 6.97 (d, J = 7.02 Hz, 1 H), 3.79 (d, J =
6.71 Hz, 1 H), 3.53 (s, 1 H), 3.18 (d, J = 4.58 Hz, 3 H), 3.00 (s,
2 H), 2.79 (d, J = 2.14 Hz, 2 H), 2.74 (s, 1 H), 2.66 (d, J = 7.93
Hz, 3 H), 1.96- 2.04 (m, 2 H), 1.91 (s, 1 H), 1.59 (s, 3 H), 1.46
(s, 1 H) 133 ##STR00647## 1.94 378.16 .sup.1H NMR (500 MHz, DMSO-
D6) .delta. ppm 12.22 (s, 1 H), 8.32 (s, 1 H), 8.01 (s, 1 H), 7.58
(d, J = 8.24 Hz, 1 H), 7.55 (s, 1 H), 7.08-7.14 (m, 1 H), 3.82 (d,
J = 9.77 Hz, 1 H), 3.56 (d, J = 9.16 Hz, 1 H), 2.96-3.04 (m, 2 H),
2.73-2.82 (m, 2 H), 2.67 (t, J = 7.48 Hz, 2 H), 2.00- 2.05 (m, 1
H), 1.91 (s, 1 H), 1.54-1.63 (m, 2 H), 1.45 (s, 1 H) 134
##STR00648## 1.86 332.16 .sup.1H NMR (500 MHz, DMSO- D6) .delta.
ppm 12.22 (s, 1 H), 8.00 (s, 1 H), 7.63 (d, J = 8.55 Hz, 1 H), 7.40
(s, 1 H), 7.00 (d, J = 7.93 Hz, 1 H), 3.82 (d, J = 9.46 Hz, 1 H),
3.56 (d, J = 9.16 Hz, 1 H), 3.00 (s, 2 H), 2.78 (s, 2 H), 2.67 (t,
J = 7.32 Hz, 2 H), 2.02 (s, 1 H), 1.91 (s, 1 H), 1.58 (s, 2 H),
1.45 (s, 1 H) 135 ##STR00649## 1.51 328.28 .sup.1H NMR (500 MHz,
DMSO- D6) .delta. ppm 11.88 (s, 2 H), 8.01 (s, 1 H), 7.48 (d, J =
8.55 Hz, 2 H), 6.73 (s, 2 H), 6.62 (dd, J = 8.70, 1.68 Hz, 2 H),
3.77- 3.83 (m, 7 H), 3.55 (d, J = 8.85 Hz, 2 H), 2.97-3.05 (m, 4
H), 2.80 (s, 3 H), 2.65-2.72 (m, 4 H), 1.98-2.04 (m, 3 H), 1.92 (s,
2 H), 1.60 (d, J = 2.14 Hz, 2 H), 1.59 (s, 2 H), 1.47 (s, 2 H) 136
##STR00650## 0.91 338.20 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 8.69 (1 H, br. s.), 8.12 (1 H, d, J = 1.51 Hz), 8.08 (1 H, d, J
= 1.26 Hz), 3.91 (1 H, d, J = 9.57 Hz), 3.57 (1 H, d, J = 9.32 Hz),
3.29 (1 H, dd, J = 14.86, 1.51 Hz), 2.62-3.01 (5 H, m), 2.01-2.18
(2 H, m), 1.61-1.76 (1 H, m), 1.39- 1.59 (2 H, m) 137 ##STR00651##
0.77 274.30 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.92 (1
H, br. s.), 8.27 (1 H, s), 7.92 (1 H, s), 3.90 (1 H, d, J = 9.32
Hz), 3.56 (1 H, d, J = 9.32 Hz), 3.32 (1 H, dd, J = 14.86, 1.76
Hz), 2.67-3.02 (5 H, m), 2.42 (3 H, s), 2.10- 2.21 (1 H, m, J =
13.17, 9.84, 3.49, 3.49, 3.49 Hz), 2.04- 2.10 (1 H, m), 1.64-1.74
(1 H, m), 1.39-1.59 (2 H, m) 138 ##STR00652## 0.84 337.40 .sup.1H
NMR (400 MHz, MeOD- d.sub.4) d ppm 9.39 (1 H, s), 8.57- 8.73 (2 H,
m), 8.52 (1 H, d, J = 5.79 Hz), 7.56 (1 H, dd, J = 8.06, 5.04 Hz),
6.54-7.00 (1 H, m), 4.11 (1 H, d, J = 10.32 Hz), 3.79 (1 H, d, J =
10.58 Hz), 3.02-3.29 (2 H, m), 2.72- 3.02 (4 H, m), 2.01-2.25 (2 H,
m), 1.47-1.89 (3 H, m) 139 ##STR00653## 1.15 309.30 .sup.1H NMR
(500 MHz, MeOD- d.sub.4) .delta. ppm 8.07 (1 H, d, J = 8.85 Hz),
7.88 (1 H, d, J = 8.24 Hz), 7.75 (1 H, d, J = 7.93 Hz), 7.56- 7.66
(1 H, m), 7.39 (1 H, t, J = 7.48 Hz), 7.06 (1 H, d, J = 8.85 Hz),
4.09 (1 H, d, J = 10.07 Hz), 3.78 (1 H, d, J = 10.07 Hz), 3.27 (1
H, d, J = 14.95 Hz), 3.06-3.18 (1 H, m), 2.93-3.01 (2 H, m), 2.75-
2.93 (2 H, m), 2.07-2.29 (2 H, m), 1.57-1.88 (3 H, m) 140
##STR00654## 1.26 327.30 .sup.1H NMR (500 MHz, MeOD- d.sub.4)
.delta. ppm 8.05 (1 H, d, J = 8.85 Hz), 7.92 (1 H, dd, J = 9.00,
5.34 Hz), 7.35-7.49 (2 H, m), 7.08 (1 H, d, J = 8.24 Hz), 4.09 (1
H, d, J = 10.07 Hz), 3.78 (1 H, d, J = 10.07 Hz), 3.22-3.30 (1 H,
m), 3.09-3.18 (1 H, m), 2.91-3.04 (2 H, m), 2.71- 2.91 (2 H, m),
2.06-2.30 (2 H, m), 1.56-1.90 (3 H, m) 141 ##STR00655## 0.80 339.40
.sup.1H NMR (400 MHz, MeOD- d.sub.4) .delta. ppm 8.58 (2 H, s),
3.99 (1 H, d, J = 10.07 Hz), 3.69 (1 H, d, J = 10.07 Hz), 3.16-3.24
(1 H, m), 3.03-3.12 (1 H, m), 2.85-2.95 (2 H, m), 2.70- 2.85 (2 H,
m), 1.96-2.21 (2 H, m), 1.42-1.87 (3 H, m) 142 ##STR00656## 0.80
294.40 .sup.1H NMR (400 MHz, MeOD- d.sub.4) .delta. ppm 8.54 (1 H,
d, J = 1.01 Hz), 6.83 (1 H, br. s.), 4.04 (1 H, d, J = 10.32 Hz),
3.73 (1 H, d, J = 10.32 Hz), 3.04-3.25 (2 H, m), 2.69-2.97 (4 H,
m), 1.96-2.18 (2 H, m), 1.43- 1.84 (3 H, m) 143 ##STR00657## 1.60
330.28 .sup.1H NMR (500 MHz, DMSO- D6) .delta. ppm 11.95 (s, 1 H),
7.54 (s, 1 H), 7.35 (s, 1 H), 7.22 (s, 1 H), 7.04 (s, 1 H), 6.09
(s, 1 H), 3.66 (s, 1 H), 3.39 (d, J = 7.02 Hz, 1 H), 3.04 (s, 1 H),
2.99 (s, 1 H), 2.84 (s, 2 H), 2.67 (s, 3 H), 2.02 (s, 1 H), 1.92
(s, 1 H), 1.59 (s, 2 H), 1.51 (s, 1 H) 144 ##STR00658## 1.00 326.30
.sup.1H NMR (400 MHz, MeOD- d.sub.4) .delta. ppm 8.47 (1 H, s),
7.30- 7.79 (1 H, m), 6.28 (1 H, br. s.), 4.02 (1 H, d, J = 10.32
Hz), 3.71 (1 H, d, J = 10.32 Hz), 3.16-3.25 (1 H, m), 3.04- 3.13 (1
H, m), 2.85-2.99 (2 H, m), 2.70-2.86 (2 H, m), 1.94- 2.20 (2 H, m),
1.49-1.84 (3 H, m). M.P. 185-90.degree. C. 145 ##STR00659## 1.16
344.30 .sup.1H NMR (400 MHz, MeOD- d.sub.4) .delta. ppm 7.06 (1 H,
s), 3.96 (1 H, d, J = 9.82 Hz), 3.65 (1 H, d, J = 10.07 Hz),
3.18-3.26 (1 H, m), 3.05-3.13 (1 H, m), 2.87- 2.98 (2 H, m),
2.70-2.86 (5 H, m), 2.57 (3 H, s), 2.03-2.20 (2 H, m), 1.51-1.85 (3
H, m) 146 ##STR00660## 1.20 309.30 .sup.1H NMR (400 MHz, MeOD-
d.sub.4) .delta. ppm 8.46 (1 H, d, J = 8.31 Hz), 8.06 (1 H, d, J =
6.04 Hz), 7.58-7.78 (2 H, m), 7.51 (1 H, td, J = 7.68, 1.26 Hz),
7.25 (1 H, d, J = 6.04 Hz), 3.99 (1 H, d, J = 9.82 Hz), 3.67 (1 H,
d, J = 9.82 Hz), 3.23 (1 H, s), 3.04- 3.13 (1 H, m), 2.89-2.99 (2
H, m), 2.71-2.88 (2 H, m), 2.04-2.27 (2 H, m), 1.46- 1.85 (3 H, m)
147 ##STR00661## 0.98 310.30 .sup.1H NMR (400 MHz, MeOD- d.sub.4)
.delta. ppm 8.72 (1 H, s), 8.46 (1 H, d, J = 7.81 Hz), 7.72-7.88 (2
H, m), 7.56 (1 H, ddd, J = 8.25, 6.86, 1.26 Hz), 4.11 (1 H, d, J =
10.32 Hz), 3.80 (1 H, d, J = 10.32 Hz), 3.09-3.17 (1 H, m), 2.97 (2
H, t, J = 7.43 Hz), 2.76-2.89 (2 H, m), 2.04- 2.26 (2 H, m),
1.50-1.87 (3 H, m) 148 ##STR00662## 0.81 285.30 .sup.1H NMR (400
MHz, MeOD- d.sub.4) .delta. ppm 8.64 (1 H, d, J = 1.26 Hz), 8.28 (1
H, d, J = 1.26 Hz), 4.19 (1 H, d, J = 10.83 Hz), 3.97 (1 H, d, J =
11.08 Hz), 3.86 (1 H, dd, J = 14.60, 1.51 Hz), 3.72 (1 H, dd, J =
14.86, 2.52 Hz), 3.44-3.56 (1 H, m), 3.31-3.43 (3 H, m), 2.49- 2.58
(1 H, m), 2.28-2.42 (1 H, m, J = 13.53, 10.07, 3.56, 3.56, 3.27
Hz), 1.91-2.18 (3 H, m) 149 ##STR00663## 0.73 290.30 .sup.1H NMR
(400 MHz, MeOD- d.sub.4) .delta. ppm 8.28 (2 H, s), 3.95 (1 H, d, J
= 9.82 Hz), 3.86 (3 H, s), 3.65 (1 H, d, J = 9.82 Hz), 3.15- 3.24
(1 H, m), 3.03-3.11 (1 H, m), 2.68-2.96 (4 H, m), 1.97-2.19 (2 H,
m), 1.49- 1.85 (3 H, m) 150 ##STR00664## 0.90 320.40 .sup.1H NMR
(400 MHz, MeOD- d.sub.4) .delta. ppm 5.87 (1 H, br. s.), 3.99 (1 H,
d, J = 10.32 Hz), 3.92 (3 H, s), 3.88 (3 H, s), 3.67 (1 H, d, J =
10.32 Hz), 3.14-3.22 (1 H, m), 3.01- 3.10 (1 H, m), 2.70-2.94 (4 H,
m), 1.98-2.16 (2 H, m), 1.51- 1.82 (3 H, m) 151 ##STR00665## 1.15
366.30 .sup.1H NMR (400 MHz, MeOD- d.sub.4) .delta. ppm 8.74 (2 H,
s), 7.53 (2 H, d, J = 8.81 Hz), 7.02 (2 H, d, J = 8.81 Hz), 4.01 (1
H, d, J = 10.07 Hz), 3.81 (3 H, s), 3.71 (1 H, d, J = 10.07 Hz),
3.26 (1 H, s), 3.08-3.18 (1 H, m), 2.73-3.03 (4 H, m), 2.16 (2 H,
br. s.), 1.51-1.90 (3 H, m) 152 ##STR00666## 1.27 350.40 .sup.1H
NMR (400 MHz, MeOD- d.sub.4) .delta. ppm 8.78 (2 H, s), 7.27- 7.47
(3 H, m), 7.20 (1 H, d, J = 7.30 Hz), 4.02 (1 H, d, J = 10.07 Hz),
3.71 (1 H, d, J = 10.07 Hz), 3.24 (1 H, d, J = 15.11 Hz), 3.03-3.14
(1 H, m), 2.70-3.00 (4 H, m), 2.40 (3 H, s), 2.03-2.22 (2 H, m),
1.53-1.84 (3 H, m) 153 ##STR00667## 2.13 362.28 .sup.1H NMR (400
MHz, MeOD- d.sub.4) .delta. ppm 8.37 (1 H, s), 8.24 (1 H, dd, J =
7.55, 1.51 Hz), 7.88 (1 H, s), 7.19-7.44 (3 H, m), 4.02 (1 H, d, J
= 10.07 Hz), 3.71 (1 H, d, J = 10.07 Hz), 3.24 (1 H, d, J = 16.87
Hz), 3.05-3.12 (1 H, m), 2.70- 2.99 (8 H, m), 2.05-2.21 (2 H, m),
1.48-1.84 (3 H, m) 154 ##STR00668## 0.57 339.10 .sup.1H NMR (500
MHz, MeOD- d.sub.4) .delta. ppm 8.07 (d, J = 5.49 Hz, 1 H),
7.47-7.71 (m, 1 H), 7.13- 7.34 (m, 1 H), 3.91-4.06 (m, 1 H),
3.56-3.73 (m, 1 H), 3.14- 3.24 (m, 1 H), 3.02-3.14 (m, 1 H),
2.73-3.01 (m, 4 H), 1.98- 2.21 (m, 2 H), 1.58-1.85 (m, 3 H) 155
##STR00669## 0.64 339.04 .sup.1H NMR (500 MHz, MeOD- d.sub.4)
.delta. ppm 7.45-7.58 (m, 1 H), 7.04-7.16 (m, 1 H), 6.90 (d, J =
7.32 Hz, 1 H), 3.95-4.09 (m, 1 H), 3.67-3.77 (m, 1 H), 3.18-3.28
(m, 1 H), 3.07- 3.17 (m, 1 H), 2.76-3.05 (m, 4 H), 2.04-2.22 (m, 2
H), 1.50- 1.89 (m, 3 H) 156 ##STR00670## 1.47 388.40 .sup.1H NMR
(500 MHz, MeOD- d.sub.4) .delta. ppm 7.96-8.03 (1 H, m), 7.88-7.95
(1 H, m), 7.75- 7.84 (1 H, m), 7.61-7.73 (1 H, m), 7.06 (1 H, d, J
= 8.55 Hz), 4.01-4.11 (1 H, m), 3.69- 3.82 (1 H, m), 3.20-3.29 (1
H, m), 3.09-3.19 (1 H, m), 2.76- 3.02 (4 H, m), 2.09-2.23 (2 H, m),
1.58-1.87 (3 H, m) 157 ##STR00671## 1.33 323.50 .sup.1H NMR (500
MHz, MeOD- d.sub.4) .delta. ppm 7.99 (1 H, d, J = 8.55 Hz), 7.77 (1
H, d, J = 8.55 Hz), 7.41-7.57 (2 H, m), 7.02 (1 H, d, J = 8.55 Hz),
4.08 (1 H, d, J = 9.77 Hz), 3.76 (1 H, d, J = 9.77 Hz), 3.26 (1 H,
d, J = 14.95 Hz), 3.08-3.17 (1 H, m), 2.78-3.02 (4 H, m), 2.49 (3
H, s), 2.08-2.31 (2 H, m), 1.54-1.89 (3 H, m) 158 ##STR00672## 1.19
316.16 .sup.1H NMR (500 MHz, DMSO- D6) .delta. ppm 12.36 (s, 1 H),
7.98 (s, 1 H), 7.19-7.27 (m, 1 H), 7.15 (d, J = 8.24 Hz, 1 H),
6.64- 6.71 (m, 1 H), 3.80 (d, J = 7.63 Hz, 1 H), 3.54 (s, 1 H),
2.95-3.04 (m, 2 H), 2.78 (s, 2 H), 2.66 (s, 2 H), 2.01 (s, 1 H),
1.93 (s, 1 H), 1.58 (s, 2 H), 1.46 (s, 1 H) 159 ##STR00673## 1.40
316.16 .sup.1H NMR (500 MHz, DMSO- D6) .delta. ppm 12.20 (s, 1 H),
7.98 (s, 1 H), 7.38 (dd, J = 8.55, 3.36 Hz, 1 H), 7.32 (d, J = 8.55
Hz, 1 H), 7.18 (t, J = 8.85 Hz, 1 H), 3.81 (d, J = 8.85 Hz, 1 H),
3.55 (d, J = 8.55 Hz, 1 H), 2.97- 3.05 (m, 2 H), 2.79 (s, 2 H),
2.68 (t, J = 7.32 Hz, 3 H), 2.02 (s, 1 H), 1.92 (s, 1 H), 1.59 (d,
J = 6.41 Hz, 2 H), 1.47 (d, J = 8.85 Hz, 1 H) 160 ##STR00674## 0.83
260.27 .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.89 (1 H, br. s.),
8.80 (1 H, d, J = 5.79 Hz), 7.53 (1 H, br. s.), 3.91 (1 H, d, J =
10.83 Hz), 3.60 (1 H, d, J = 10.83 Hz), 3.16-3.25 (1 H, m), 3.02-
3.12 (1 H, m), 2.68-2.99 (4 H, m), 1.94-2.17 (2 H, m), 1.53-1.83 (3
H, m) 161 ##STR00675## 1.55 328.21 .sup.1H NMR (400 MHz, MeOD-
d.sub.4) .delta. ppm 8.35 (1 H, s), 4.01 (1 H, d, J = 10.07 Hz),
3.70 (1 H, d, J = 10.07 Hz), 3.20-3.27 (1 H, m), 3.05-3.13 (1 H,
m), 2.69-2.99 (4 H, m), 2.34 (3 H, s), 2.29 (3 H, s), 2.01-2.22 (2
H, m), 1.51-1.83 (3 H, m) 162 ##STR00676## 1.85 354.25 .sup.1H NMR
(500 MHz, DMSO- D6) .delta. ppm 11.75-12.09 (m, J = 14.95 Hz, 1 H),
7.62 (d, J = 7.93 Hz, 2 H), 7.38-7.54 (m, J = 10.99 Hz, 1 H), 6.95
(d, J = 6.71 Hz, 2 H), 6.13 (s, 1 H), 3.76-3.83 (m, 4 H), 3.74 (s,
1 H), 3.38 (d, J = 2.75 Hz, 1 H), 2.98 (s, 1 H), 2.82 (s, 2 H),
2.62-2.71 (m, 2 H), 2.00 (s, 1 H), 1.92 (s, 1 H), 1.58 (s, 2 H),
1.50 (s, 1 H) 163 ##STR00677## 2.09 366.17 .sup.1H NMR (500 MHz,
DMSO- D6) .delta. ppm 12.54 (s, 1 H), 8.05 (s, 1 H), 7.97 (s, 1 H),
7.52- 7.60 (m, 2 H), 3.82 (s, 1 H), 3.58 (s, 1 H), 3.01 (s, 2 H),
2.78 (s, 2 H), 2.64-2.70 (m, 2 H), 1.99-2.05 (m, 1 H), 1.92 (s, 1
H), 1.58 (s, 2 H), 1.47 (s, 1 H)
164 ##STR00678## 2.12 366.17 .sup.1H NMR (500 MHz, DMSO- D6)
.delta. ppm 12.54 (s, 1 H), 8.03 (s, 1 H), 7.85 (d, J = 8.24 Hz, 1
H), 7.70 (s, 1 H), 7.26 (d, J = 8.24 Hz, 1 H), 3.83 (d, J = 9.46
Hz, 1 H), 3.57 (d, J = 8.85 Hz, 1 H), 2.97-3.05 (m, 2 H), 2.79 (s,
2 H), 2.67 (t, J = 7.48 Hz, 2 H), 2.03 (d, J = 2.44 Hz, 1 H), 1.92
(s, 1 H), 1.59 (d, J = 5.80 Hz, 2 H), 1.46 (s, 1 H) 165
##STR00679## 1.24 298.16 .sup.1H NMR (500 MHz, DMSO- D6) .delta.
ppm 12.07 (s, 1 H), 8.02 (s, 1 H), 7.63 (d, J = 7.63 Hz, 1 H),
7.27-7.36 (m, 2 H), 6.99 (t, J = 7.32 Hz, 1 H), 3.82 (d, J = 9.46
Hz, 1 H), 3.56 (d, J = 8.85 Hz, 1 H), 3.00 (s, 2 H), 2.79 (s, 2 H),
2.67 (t, J = 6.87 Hz, 2 H), 2.02 (d, J = 2.75 Hz, 1 H), 1.93 (s, 1
H), 1.58 (s, 2 H), 1.47 (s, 1 H) 166 ##STR00680## 1.16 366.30
.sup.1H NMR (500 MHz, MeOD- d.sub.4) .delta. ppm 8.53 (1 H, d, J =
5.19 Hz), 8.02-8.15 (2 H, m), 7.39 (1 H, d, J = 5.19 Hz), 7.01-
7.11 (2 H, m), 4.06 (1 H, d, J = 10.07 Hz), 3.89 (3 H, s), 3.74 (1
H, d, J = 10.07 Hz), 3.28 (1 H, d, J = 14.95 Hz), 3.13 (1 H, d, J =
14.95 Hz), 2.73-3.02 (4 H, m), 2.07- 2.25 (2 H, m), 1.54-1.86 (3 H,
m) 167 ##STR00681## 1.25 344.40 .sup.1H NMR (500 MHz, MeOD-
d.sub.4) .delta. ppm 8.48 (1 H, s), 4.05 (1 H, d, J = 10.07 Hz),
3.74 (1 H, d, J = 10.07 Hz), 3.26 (1 H, s), 3.14 (1 H, d, J = 14.65
Hz), 2.76-3.02 (4 H, m), 2.59 (3 H, s), 2.46 (3 H, s), 2.09-2.23 (2
H, m), 1.57-1.89 (3 H, m) 168 ##STR00682## 1.73 390.20 .sup.1H NMR
(500 MHz, MeOD- d.sub.4) .delta. ppm 9.27 (1 H, s), 8.14 (1 H, d, J
= 2.14 Hz), 7.93 (1 H, dd, J = 8.85, 1.83 Hz), 7.78 (1 H, d, J =
8.85 Hz), 4.11 (1 H, d, J = 10.07 Hz), 3.80 (1 H, d, J = 10.07 Hz),
3.28 (1 H, s), 3.15 (1 H, d, J = 14.95 Hz), 2.78-3.02 (4 H, m),
2.05- 2.25 (2 H, m), 1.52-1.88 (3 H, m) 169 ##STR00683## 0.71
260.40 .sup.1H NMR (500 MHz, MeOD- d.sub.4) .delta. ppm 8.73 (1 H,
s), 8.59 (2 H, br. s.), 3.86 (1 H, d, J = 9.77 Hz), 3.56 (1 H, d, J
= 9.77 Hz), 3.18-3.26 (1 H, m), 3.05- 3.12 (1 H, m), 2.72-2.99 (4
H, m), 2.17 (1 H, br. s.), 2.01 (1 H, br. s.), 1.54-1.86 (3 H, m)
170 ##STR00684## 0.78 290.30 .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 8.57 (1 H, br. s.), 7.97 (1 H, d, J = 1.51 Hz), 7.80 (1
H, d, J = 1.26 Hz), 3.90 (3 H, s), 3.87 (1 H, d, J = 9.06 Hz), 3.54
(1 H, d, J = 9.06 Hz), 3.32 (1 H, dd, J = 14.86, 1.76 Hz),
2.69-3.04 (5 H, m), 2.13-2.23 (1 H, m), 2.06-2.11 (1 H, m), 1.63-
1.76 (1 H, m), 1.41-1.61 (2 H, m) 171 ##STR00685## 1.06 368.40
.sup.1H NMR (400 MHz, MeOD- d.sub.4) .delta. ppm 3.99 (1 H, d, J =
10.07 Hz), 3.68 (1 H, d, J = 10.07 Hz), 3.15-3.24 (1 H, m),
3.02-3.10 (1 H, m), 2.68- 2.96 (4 H, m), 2.53 (6 H, s), 2.02-2.19
(2 H, m), 1.46- 1.83 (3 H, m) 172 ##STR00686## 0.89 304.25 .sup.1H
NMR (400 MHz, MeOD- d.sub.4) .delta. ppm 6.22 (1 H, s), 3.99 (1 H,
d, J = 10.07 Hz), 3.89 (3 H, s), 3.67 (1 H, d, J = 10.07 Hz),
3.15-3.25 (1 H, m), 3.00- 3.11 (1 H, m), 2.67-2.97 (4 H, m), 2.32
(3 H, s), 2.00-2.19 (2 H, m), 1.42-1.83 (3 H, m) 173 ##STR00687##
1.07 371.17 .sup.1H NMR (500 MHz, DMSO- D6) .delta. ppm 8.57 (s, 1
H), 7.46- 7.54 (m, 6 H), 7.31 (t, J = 7.93 Hz, 2 H), 6.87 (dd, J =
8.24, 2.44 Hz, 2 H), 3.87 (d, J = 9.77 Hz, 2 H), 3.82 (s, 6 H),
3.61 (d, J = 9.77 Hz, 2 H), 3.03 (s, 4 H), 2.75-2.84 (m, 4 H), 2.68
(t, J = 7.63 Hz, 4 H), 2.06 (s, 2 H), 1.93 (s, 2 H), 1.56-1.64 (m,
4 H), 1.50 (dd, J = 9.46, 2.44 Hz, 2 H) 174 ##STR00688## 1.32
363.44 .sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 8.73-8.93
(m, 1 H), 7.73 (s, 1 H), 7.20 (s, 1 H), 3.91 (d, J = 9.77 Hz, 1 H),
3.66 (d, J = 10.07 Hz, 1 H), 3.05 (s, 2 H), 2.74-2.89 (m, 1 H),
2.68 (t, J = 7.63 Hz, 1 H), 2.54 (s, 3 H), 2.09 (br. s., 2 H),
1.83- 1.98 (m, 1 H), 1.42-1.69 (m, 2 H) 175 ##STR00689## 0.40
274.26 .sup.1H NMR (400 MHz, MeOD- d.sub.4) .delta. ppm 8.36 (1 H,
d, J = 5.04 Hz), 6.82 (1 H, d, J = 5.04 Hz), 3.98 (1 H, d, J =
10.07 Hz), 3.67 (1 H, d, J = 10.07 Hz), 3.17-3.25 (1 H, m), 3.03-
3.10 (1 H, m), 2.70-2.96 (4 H, m), 2.41 (3 H, s), 1.98-2.22 (2 H,
m), 1.44-1.87 (3 H, m) 176 ##STR00690## 0.72 288.25 .sup.1H NMR
(400 MHz, MeOD- d.sub.4) .delta. ppm 6.72 (1 H, s), 3.98 (1 H, d, J
= 10.07 Hz), 3.67 (1 H, d, J = 9.82 Hz), 3.18-3.24 (1 H, m),
3.02-3.12 (1 H, m), 2.65-2.96 (4 H, m), 2.36 (6 H, s), 2.02-2.21 (2
H, m), 1.45- 1.83 (3 H, m) 177 ##STR00691## 2.13 362.28 .sup.1H NMR
(400 MHz, MeOD- d.sub.4) .delta. ppm 8.37 (1 H, s), 8.24 (1 H, dd,
J = 7.55, 1.51 Hz), 7.88 (1 H, s), 7.19-7.44 (3 H, m), 4.02 (1 H,
d, J = 10.07 Hz), 3.71 (1 H, d, J = 10.07 Hz), 3.24 (1 H, d, J =
16.87 Hz), 3.05-3.12 (1 H, m), 2.70- 2.99 (8 H, m), 2.05-2.21 (2 H,
m), 1.48-1.84 (3 H, m) 178 ##STR00692## 1.31 370.18 .sup.1H NMR
(400 MHz, MeOD- d.sub.4) .delta. ppm 8.32 (1 H, s), 3.94- 4.03 (4
H, m), 3.68 (1 H, d, J = 10.07 Hz), 3.15-3.26 (1 H, m), 3.01-3.12
(1 H, m), 2.68- 2.96 (4 H, m), 2.01-2.16 (2 H, m), 1.51-1.84 (3 H,
m) 179 ##STR00693## 1.87 353.28 .sup.1H NMR (500 MHz, MeOD-
d.sub.4) .delta. ppm 7.83 (1 H, s), 7.63- 7.76 (1 H, m), 7.19 (1 H,
dd, J = 9.00, 2.90 Hz), 7.09 (1 H, d, J = 2.75 Hz), 3.93-4.02 (1 H,
m), 3.84-3.93 (3 H, m), 3.69 (1 H, d, J = 9.77 Hz), 3.26 (1 H, d, J
= 14.65 Hz), 3.11 (1 H, d, J = 14.95 Hz), 2.70-3.01 (4 H, m), 2.39
(3 H, s), 2.06-2.27 (2 H, m), 1.57-1.85 (3 H, m) 180 ##STR00694##
1.94 350.30 .sup.1H NMR (400 MHz, MeOD- d.sub.4) .delta. ppm
7.91-8.15 (2 H, m), 7.38-7.53 (3 H, m), 7.31 (1 H, s), 4.02 (1 H,
d, J = 10.07 Hz), 3.70 (1 H, d, J = 10.07 Hz), 3.17-3.26 (1 H, m),
3.02- 3.14 (1 H, m), 2.66-2.98 (4 H, m), 2.48 (3 H, s), 2.03-2.20
(2 H, m), 1.49-1.83 (3 H, m) 181 ##STR00695## 1.17 336.40 .sup.1H
NMR (400 MHz, MeOD- d.sub.4) .delta. ppm 8.55 (1 H, d, J = 5.29
Hz), 8.08 (2 H, dd, J = 6.67, 2.90 Hz), 7.44-7.53 (3 H, m), 7.40 (1
H, d, J = 5.29 Hz), 4.01 (1 H, d, J = 10.07 Hz), 3.70 (1 H, d, J =
10.07 Hz), 3.20-3.28 (1 H, m), 3.04-3.14 (1 H, m), 2.67-2.99 (4 H,
m), 2.05- 2.24 (2 H, m), 1.49-1.85 (3 H, m) 182 ##STR00696## 0.42
289.22 .sup.1H NMR (400 MHz, MeOD- d.sub.4) .delta. ppm 4.02 (1 H,
d, J = 10.32 Hz), 3.71 (1 H, d, J = 10.32 Hz), 3.23 (1 H, d, J =
15.36 Hz), 3.03-3.13 (1 H, m), 2.69-2.98 (4 H, m), 2.54 (3 H, s),
2.48 (3 H, s), 2.01- 2.21 (2 H, m), 1.41-1.85 (3 H, m) 183
##STR00697## 1.19 382.19 .sup.1H NMR (500 MHz, DMSO- D6) .delta.
ppm 12.37 (s, 1 H), 8.01 (s, 1 H), 7.53 (s, 1 H), 7.45 (d, J = 9.16
Hz, 1 H), 7.28 (d, J = 8.85 Hz, 1 H), 3.82 (d, J = 9.16 Hz, 1 H),
3.57 (d, J = 9.46 Hz, 1 H), 3.00 (s, 2 H), 2.78 (s, 2 H), 2.67 (t,
J = 7.48 Hz, 2 H), 1.96-2.04 (m, 1 H), 1.91 (d, J = 8.85 Hz, 1 H),
1.55- 1.63 (m, 2 H), 1.47 (d, J = 7.02 Hz, 1 H) 184 ##STR00698##
0.75 274.30 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.08 (1
H, br. s.), 8.12 (1 H, s), 7.85 (1 H, s), 3.89 (1 H, d, J = 9.32
Hz), 3.55 (1 H, d, J = 9.32 Hz), 3.28 (1 H, dd, J = 14.86, 1.26
Hz), 2.62-2.97 (5 H, m), 2.35 (3 H, s), 2.06- 2.17 (1 H, m, J =
13.13, 9.82, 3.53, 3.38, 3.38 Hz), 2.04 (1 H, br. s.), 1.59-1.73 (1
H, m), 1.37-1.57 (2 H, m) 185 ##STR00699## 0.75 274.30 .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. ppm 9.29 (1 H, br. s.), 7.90 (2 H,
br. s.), 3.90 (1 H, d, J = 9.32 Hz), 3.55 (1 H, d, J = 9.06 Hz),
3.35 (1 H, d, J = 14.86 Hz), 2.65-3.08 (5 H, m), 2.56 (3 H, s),
2.12-2.27 (1 H, m), 2.08 (1 H, br. s.), 1.62- 1.77 (1 H, m),
1.38-1.61 (2 H, m) 186 ##STR00700## 0.79 288.30 .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. ppm 9.16 (1 H, br. s.), 7.78 (1 H, s),
3.88 (1 H, d, J = 9.06 Hz), 3.53 (1 H, d, J = 9.32 Hz), 3.34 (1 H,
dd, J = 14.86, 2.01 Hz), 2.64-3.07 (5 H, m), 2.55 (3 H, s), 2.38 (3
H, s), 2.14- 2.25 (1 H, m), 2.04-2.11 (1 H, m), 1.68 (1 H, dddd, J
= 13.94, 9.66, 4.53, 4.34 Hz), 1.41- 1.61 (2 H, m) 187 ##STR00701##
0.58 293.06 .sup.1H NMR (500 MHz, DMSO- d.sub.6) d ppm 3.73-3.87
(m, 1 H), 3.47-3.58 (m, 1 H), 2.98 (s, 2 H), 2.71-2.85 (m, 2 H),
2.65 (t, J = 7.78 Hz, 2 H), 2.18 (s, 3 H), 2.11 (s, 3 H), 2.00 (br.
s., 1 H), 1.80-1.96 (m, 1 H), 1.57 (dd, J = 8.24, 2.75 Hz, 3 H) 188
##STR00702## 0.86 325.20 .sup.1H NMR (500 MHz, DMSO- d.sub.6)
.delta. ppm 8.07 (d, J = 7.32 Hz, 2 H), 7.42 (d, J = 7.63 Hz, 3 H),
3.87 (s, 1 H), 3.55-3.70 (m, 1 H), 3.02 (s, 2 H), 2.72-2.88 (m, 2
H), 2.68 (s, 2 H), 1.98- 2.10 (m, 1 H), 1.81-1.97 (m, 1 H),
1.40-1.68 (m, 3 H) 189 ##STR00703## 1.04 355.24 .sup.1H NMR (500
MHz, DMSO- d.sub.6) .delta. ppm 7.36-7.47 (m, 3 H), 7.24-7.34 (m, 2
H), 3.84 (d, J = 10.07 Hz, 1 H), 3.58 (d, J = 9.77 Hz, 1 H), 3.01
(s, 2 H), 2.71-2.87 (m, 2 H), 2.66 (t, J = 7.63 Hz, 2 H), 2.32 (s,
3 H), 2.04 (br. s., 1 H), 1.83-1.98 (m, 1 H), 1.43-1.64 (m, 3 H)
190 ##STR00704## 0.26 262.11 .sup.1H NMR (500 MHz, DMSO- d.sub.6) d
ppm 7.43 (br. s., 1 H), 5.56-5.76 (m, 1 H), 3.58- 3.83 (m, 4 H),
3.37-3.52 (m, 1 H), 2.94 (s, 2 H), 2.68-2.88 (m, 2 H), 2.60-2.68
(m, 2 H), 1.94 (br. s., 2 H), 1.37-1.65 (m, 3 H) 191 ##STR00705##
1.65 350.30 .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.43 (1 H, s),
7.58 (2 H, dd, J = 7.55, 1.76 Hz), 7.39- 7.52 (3 H, m), 3.96 (1 H,
d, J = 10.07 Hz), 3.66 (1 H, d, J = 9.82 Hz), 3.25 (1 H, d, J =
16.12 Hz), 3.05-3.14 (1 H, m), 2.88-2.98 (2 H, m), 2.68- 2.87 (2 H,
m), 2.24 (3 H, s), 2.03-2.17 (2 H, m), 1.45- 1.84 (3 H, m) 192
##STR00706## 1.02 337.28 .sup.1H NMR (400 MHz, MeOD) .delta. ppm
8.64 (2 H, d, J = 5.29 Hz), 8.40 (1 H, d, J = 7.81 Hz), 7.86- 7.97
(1 H, m), 7.80 (1 H, d, J = 5.04 Hz), 7.47 (1 H, dd, J = 6.55, 5.04
Hz), 4.03 (1 H, d, J = 10.07 Hz), 3.74 (1 H, d, J = 10.07 Hz), 3.29
(1 H, d, J = 1.51 Hz), 3.12-3.22 (1 H, m), 2.99 (2 H, t, J = 7.68
Hz), 2.78-2.92 (2 H, m), 2.03- 2.26 (2 H, m), 1.45-1.88 (3 H, m)
193 ##STR00707## NA NA .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.69
(1 H, s), 7.61-7.78 (2 H, m), 7.34-7.56 (3 H, m), 3.99 (1 H, d, J =
10.07 Hz), 3.68 (1 H, d, J = 10.32 Hz), 3.15-3.25 (1 H, m), 3.02-
3.11 (1 H, m), 2.92 (2 H, t, J = 7.55 Hz), 2.69-2.86 (2 H, m),
2.01-2.20 (2 H, m), 1.47- 1.81 (3 H, m) 194 ##STR00708## NA NA
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.24 (1 H, d, J = 1.51 Hz),
8.60-8.68 (2 H, m), 8.53 (1 H, dt, J = 7.99, 1.92 Hz), 7.40- 7.64
(2 H, m), 4.05 (1 H, d, J = 10.32 Hz), 3.76 (1 H, d, J = 10.07 Hz),
3.33-3.40 (1 H, m), 3.17-3.26 (1 H, m), 2.82- 3.09 (4 H, m),
2.09-2.31 (2 H, m), 1.55-1.91 (3 H, m) 195 ##STR00709## 1.41 403.36
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.65 (1 H, s), 8.40 (1 H,
s), 8.17 (1 H, s), 7.25-7.47 (5 H, m), 4.04 (1 H, d, J = 10.32 Hz),
3.73 (1 H, d, J = 10.32 Hz), 3.17-3.27 (1 H, m), 3.04- 3.13 (1 H,
m), 2.63-3.00 (4 H, m), 2.00-2.22 (2 H, m), 1.48-1.87 (3 H, m) 196
##STR00710## 1.69 364.36 .sup.1H NMR (400 MHz, MeOD) .delta. ppm
8.33 (1 H, s), 8.16 (1 H, dd, J = 7.81, 1.51 Hz), 7.30- 7.44 (1 H,
m), 7.08 (1 H, t, J = 7.55 Hz), 6.95 (1 H, d, J = 8.31 Hz), 5.16 (2
H, s), 4.02 (1 H, d, J = 9.82 Hz), 3.71 (1 H, d, J = 10.07 Hz),
3.18-3.26 (1 H, m), 3.04-3.13 (1 H, m), 2.93 (2 H, t, J = 7.68 Hz),
2.71- 2.86 (2 H, m), 2.01-2.23 (2 H, m), 1.50-1.83 (3 H, m) 197
##STR00711## 0.53 277.13 .sup.1H NMR (500 MHz, DMSO- D6) .delta.
ppm 8.76 (s, 1 H), 8.15 (d, J = 2.44 Hz, 1 H), 7.55 (s, 1 H), 6.83
(s, 1 H), 3.79 (s, 1 H), 3.54 (d, J = 10.07 Hz, 1 H), 2.92-3.00 (m,
3 H), 2.70- 2.80 (m, 3 H), 2.66 (t, J = 7.78 Hz, 3 H), 1.97 (s, 1
H), 1.88 (s, 1 H), 1.53-1.61 (m, 3 H), 1.41- 1.49 (m, 1 H) 198
##STR00712## 0.77 337.07 .sup.1H NMR (500 MHz, DMSO- d.sub.6)
.delta. ppm 8.26 (br. s., 1 H), 7.76 (br. s., 1 H), 6.75 (br. s., 1
H), 3.81 (br. s., 1 H), 3.56 (d, J = 9.46 Hz, 1 H), 2.85-3.10 (m, 2
H), 2.56-2.85 (m, 4 H), 1.97 (br. s., 2 H), 1.36-1.67 (m, 3 H) 199
##STR00713## 0.16 289.19 .sup.1H NMR (500 MHz, MeOD) .delta. ppm
7.90 (d, J = 6.10 Hz, 1 H), 6.74-7.00 (m, 2 H), 3.93- 4.09 (m, 1
H), 3.88 (s, 3 H), 3.61-3.71 (m, 1 H), 3.14- 3.25 (m, 1 H), 3.06
(d, J = 1.83 Hz, 1 H), 2.69-2.99 (m, 3 H), 2.07 (br. s., 2 H), 1.74
(d, J = 8.24 Hz, 3 H), 1.18 (d, J = 6.41 Hz, 1 H) 200 ##STR00714##
0.82 332.97 .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. ppm 7.71
(dd, J = 8.78, 2.76 Hz, 1 H), 7.59 (dd, J = 8.91, 4.89 Hz, 1 H),
7.17 (td, J = 9.03, 2.76 Hz, 1 H), 3.89 (d J = 10.04 Hz, 1 H), 3.64
(d, J = 10.04 Hz, 1 H), 2.97-3.13 (m, 2 H), 2.75- 2.88 (m, 2 H),
2.67 (t, J = 7.65 Hz, 2 H), 2.02-2.12 (m, 1 H), 1.81-1.98 (m, 1 H),
1.36- 1.69 (m, 3 H) 201 ##STR00715## 1.16 314.35 .sup.1H NMR (400
MHz, MeOD) .delta. ppm 8.20 (1 H, s), 3.96 (1 H, d, J = 10.07 Hz),
3.65 (1 H, d, J = 9.82 Hz), 3.20 (1 H, d), 3.06 (1 H, d), 2.91 (2
H, t, J = 7.43 Hz), 2.71-2.84 (4 H, m), 2.66 (2 H, t, J = 6.04 Hz),
1.98-2.21 (2 H, m), 1.43-1.94 (7 H, m) 202 ##STR00716## 0.69 300.29
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.30 (1 H, s), 3.97 (1 H,
d, J = 9.82 Hz), 3.66 (1 H, d, J = 10.07 Hz), 3.21 (1 H, d), 3.06
(1 H, d), 2.74-2.97 (8 H, m), 2.11 (4 H, dq, J = 7.68, 7.51 Hz),
1.39-1.85 (3 H, m) 203 ##STR00717## 1.19 322.25 .sup.1H NMR (400
MHz, MeOD) .delta. ppm 4.00 (1 H, d, J = 10.07 Hz), 3.69 (1 H, d, J
= 10.07 Hz), 3.22 (1 H, d, J = 14.60 Hz), 3.03-3.14 (1 H, m), 2.69-
2.97 (4 H, m), 2.46 (3 H, s), 2.26 (3 H, s), 2.03-2.17 (2 H, m),
1.51-1.84 (3 H, m) 204 ##STR00718## 0.67 328.28 .sup.1H NMR (400
MHz, MeOD) .delta. ppm 8.92 (1 H, s), 4.05 (1 H, d, J = 10.32 Hz),
3.75 (1 H, d, J = 10.32 Hz), 3.24 (1 H, d, J = 1.26 Hz), 3.06-3.17
(1 H, m), 2.75-3.03 (6 H, m), 2.56- 2.68 (2 H, m), 2.13 (4 H, ddd,
J = 12.72, 6.30, 6.17 Hz), 1.54- 1.84 (3 H, m) 205 ##STR00719##
0.15 284.10 .sup.1H NMR (500 MHz, DMSO- D6) .delta. ppm 9.01 (s, 1
H), 8.40 (d, J = 4.88 Hz, 1 H), 7.21 (s, 1 H), 7.16 (s, 1 H), 3.84
(s, 1 H), 3.59 (d, J = 10.07 Hz, 1 H), 2.94-3.02 (m, 3 H), 2.70-
2.79 (m, 3 H), 2.62-2.68 (m, 3 H), 1.99 (s, 1 H), 1.87 (s, 1 H),
1.57 (d, J = 3.05 Hz, 2 H), 1.56 (s, 1 H), 1.41-1.49 (m, 1 H)
206 ##STR00720## 0.78 366.10 .sup.1H NMR (500 MHz, DMSO- D6)
.delta. ppm 12.63 (s, 1 H), 7.95 (d, J = 7.63 Hz, 2 H), 7.68 (d, J
= 6.71 Hz, 1 H), 7.16 (t, J = 7.32 Hz, 1 H), 3.84 (s, 1 H), 3.60
(s, 1 H), 3.03 (s, 2 H), 2.80 (s, 2 H), 2.64-2.72 (m, 2 H), 2.05
(s, 1 H), 1.92 (s, 1 H), 1.60 (d, J = 8.24 Hz, 2 H), 1.48 (s, 1 H)
207 ##STR00721## 0.77 335.17 .sup.1H NMR (500 MHz, DMSO- D6)
.delta. ppm 9.05 (s, 1 H), 8.52 (d, J = 2.44 Hz, 1 H), 7.91 (s, 1
H), 7.66 (d, J = 7.63 Hz, 2 H), 7.47 (t, J = 7.63 Hz, 2 H), 7.36
(t, J = 7.32 Hz, 1 H), 6.87 (s, 1 H), 3.84 (s, 1 H), 3.59 (d, J =
10.07 Hz, 1 H), 2.99 (s, 2 H), 2.72-2.81 (m, 2 H), 2.63- 2.71 (m, 3
H), 1.99 (s, 1 H), 1.90 (s, 1 H), 1.59 (s, 2 H), 1.42-1.50 (m, 1 H)
208 ##STR00722## 0.63 342.25 .sup.1H NMR (500 MHz, DMSO- D6)
.delta. ppm 9.30 (s, 1 H), 7.19 (d, J = 8.55 Hz, 1 H), 6.94 (d, J =
2.44 Hz, 1 H), 6.69 (dd, J = 8.55, 2.14 Hz, 1 H), 3.92 (d, J =
10.07 Hz, 1 H), 3.75 (s, 3 H), 3.66 (d, J = 9.77 Hz, 1 H), 3.54 (s,
3 H), 2.99-3.07 (m, 2 H), 2.79 (t, J = 7.78 Hz, 2 H), 2.68 (t, J =
7.78 Hz, 2 H), 2.02- 2.07 (m, 1 H), 1.89 (dd, J = 6.41, 3.36 Hz, 1
H), 1.56- 1.64 (m, 2 H), 1.44-1.52 (m, 1 H) 209 ##STR00723## 0.96
300.34 .sup.1H NMR (400 MHz, MeOD) d ppm 8.27 (1 H, d, J = 5.04
Hz), 6.78 (1 H, d, J = 5.29 Hz), 3.98 (1 H, d, J = 10.07 Hz), 3.66
(1 H, d, J = 10.07 Hz), 3.21 (1 H, d), 3.06 (1 H, d), 2.64-2.99 (4
H, d, J = 5.29 H 3.98 1.92-2.05 (1 H, m), 1.54- 1.82 (3 H, m),
0.92-1.11 (4 H, m) 210 ##STR00724## 1.64 378.23 .sup.1H NMR (400
MHz, MeOD) d ppm 8.42 (1 H, s), 3.97 (1 H, d, J = 10.07 Hz), 3.66
(1 H, d, J = 10.07 Hz), 3.20 (1 H, d), 3.06 (1 H, d), 2.63-2.97 (4
H, m), 2.35-2.50 (1 H, m), 1.99- 2.16 (2 H, m), 1.50-1.83 (3 H, m),
0.98-1.24 (4 H, m) 211 ##STR00725## 0.64 342.18 .sup.1H NMR (500
MHz, DMSO- D6) .delta. ppm 7.98 (s, 1 H), 7.46 (t, J = 7.93 Hz, 1
H), 6.91 (d, J = 5.49 Hz, 1 H), 6.59-6.65 (m, 1 H), 3.82-3.88 (m, 6
H), 3.82 (s, 2 H), 3.55 (s, 1 H), 3.00 (s, 2 H), 2.78 (s, 2 H),
2.67 (d, J = 6.71 Hz, 2 H), 2.01 (s, 1 H), 1.91 (s, 1 H), 1.59 (s,
2 H), 1.46 (s, 1 H) 212 ##STR00726## 0.63 342.25 .sup.1H NMR (500
MHz, DMSO- D6) .delta. ppm 7.94 (s, 1 H), 7.37 (d, J = 9.16 Hz, 1
H), 7.02 (d, J = 9.46 Hz, 1 H), 6.99 (d, J = 1.83 Hz, 1 H), 3.87
(s, 3 H), 3.82 (d, J = 9.46 Hz, 1 H), 3.78 (s, 3 H), 3.56 (d, J =
9.16 Hz, 1 H), 3.00 (s, 2 H), 2.78 (s, 2 H), 2.63-2.70 (m, 2 H),
2.01 (s, 1 H), 1.91 (s, 1 H), 1.58 (s, 2 H), 1.46 (s, 1 H) 213
##STR00727## 0.35 294.25 .sup.1H NMR (400 MHz, MeOD) d ppm 8.51 (2
H, s), 4.00 (1 H, d, J = 10.32 Hz), 3.69 (1 H, d, J = 10.32 Hz),
3.22 (1 H, d), 3.08 (1 H, d), 2.70-2.98 (4 H, m), 2.01-2.21 (2 H,
m), 1.50- 1.84 (3 H, m) 214 ##STR00728## 0.36 293.14 .sup.1H NMR
(500 MHz, DMSO- D6) .delta. ppm 8.80 (s, 1 H), 8.19 (s, 1 H), 7.66
(s, 1 H), 6.80 (s, 1 H), 3.81 (s, 1 H), 3.56 (d, J = 10.07 Hz, 1
H), 2.92-3.01 (m, 2 H), 2.70-2.79 (m, 2 H), 2.62-2.68 (m, 2 H),
1.97 (s, 1 H), 1.87 (s, 1 H), 1.57 (d, J = 7.32 Hz, 2 H), 1.40-1.49
(m, 1 H) 215 ##STR00729## 0.35 293.14 .sup.1H NMR (500 MHz, DMSO-
D6) .delta. ppm 9.01 (s, 1 H), 8.17 (d, J = 5.49 Hz, 2 H), 6.97 (s,
2 H), 6.83 (s, 1 H), 3.83 (s, 2 H), 3.57 (d, J = 10.38 Hz, 2 H),
2.93-3.01 (m, 3 H), 2.74- 2.82 (m, 3 H), 2.73 (s, 1 H), 2.66 (t, J
= 7.63 Hz, 3 H), 1.98 (s, 2 H), 1.87 (s, 2 H), 1.53- 1.62 (m, 3 H),
1.41-1.49 (m, 2 H) 216 ##STR00730## 0.82 318.30 .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. ppm 8.56 (1 H, br. s.), 7.94 (1 H, d, J =
1.26 Hz), 7.72 (1 H, d, J = 1.26 Hz), 5.14 (1 H, spt, J = 6.13 Hz),
3.85 (1 H, d, J = 9.06 Hz), 3.52 (1 H, d, J = 9.06 Hz), 3.30 (1 H,
dd, J = 14.86, 1.51 Hz), 2.67-3.03 (5 H, m), 2.11-2.26 (1 H, m, J =
13.17, 9.84, 3.65, 3.42, 3.42 Hz), 2.07 (1 H, br. s.), 1.62- 1.75
(1 H, m), 1.41-1.59 (2 H, m), 1.30 (6 H, d, J = 6.30 Hz) 217
##STR00731## 0.18 342.19 .sup.1H NMR (400 MHz, DMSO- D6) .delta.
ppm 8.09 (s, 1 H), 7.53 (s, 1 H), 3.80 (s, 1 H), 3.54 (d, J = 10.32
Hz, 1 H), 3.07-3.18 (m, 1 H), 2.91-3.01 (m, 3 H), 2.75 (d, J = 8.31
Hz, 2 H), 2.60- 2.71 (m, 2 H), 2.19 (q, J = 8.90 Hz, 1 H),
2.01-2.12 (m, 4 H), 1.94 (s, 1 H), 1.83 (d, J = 15.11 Hz, 2 H),
1.56 (s, 3 H), 1.43 (s, 1 H)
Example 218
(R)--N-(6-(methoxymethyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00732##
[0389] Step A: 6-(Methoxymethyl)pyrimidin-4-ol
##STR00733##
[0391] To a solution of methyl 4-methoxy-3-oxobutanoate (3.54 mL,
26.5 mmol) in methanol (30 ml) was added formamidine acetate (3.07
g, 29.2 mmol) and sodium methoxide (13 mL, 58.4 mmol). The mixture
was then heated to reflux for 18 hours and cooled to ambient
temperature, then concentrated. The residue was taken up in water
and the pH adjusted to 7 with 1N HCl. The aqueous mixture was
extracted with chloroform. The combined organic layers were dried
over magnesium sulfate, filtered and concentrated to afford
6-(methoxymethyl)pyrimidin-4-ol (1.38 g, 9.85 mmol, 37.1% yield).
MS (LC/MS) R.T.=0.19; [M+H].sup.+=141.20.
Step B: 4-Chloro-6-(methoxymethyl)pyrimidine
##STR00734##
[0393] 6-(Methoxymethyl)pyrimidin-4-ol (1.38 g, 9.85 mmol) was
taken up in dichloromethane (14 ml) and phosphorous oxychloride (9
mL, 97 mmol) was added at ambient temperature. The mixture was
stirred at ambient for 18 h and concentrated. The residue was taken
up in ice-water and the pH was adjusted to 7 with 1N sodium
hydroxide. The mixture was extracted with chloroform and dried over
sodium sulfate, filtered and concentrated in vacuo. The residue was
purified by column chromatography (5-10% ethyl acetate/chloroform)
to afford 4-chloro-6-(methoxymethyl)pyrimidine (1.2 g, 7.57 mmol,
77% yield) as a pale yellow oil, which solidified on standing.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.88 (1H, d, J=1.01
Hz), 7.52 (1H, d, J=1.01 Hz), 4.53 (2H, s), 3.50 (3H, s). MS
(LC/MS) R.T.=0.98; [M+H].sup.+=159.10.
Step C: 6-(Methoxymethyl)pyrimidin-4-amine
##STR00735##
[0395] A mixture of 4-chloro-6-(methoxymethyl)pyrimidine (1.2 g,
7.57 mmol) and ammonium hydroxide (20 ml) was heated in a sealed
tube for 3 hours. The mixture was cooled to ambient temperature and
concentrated. The residue was triturated with ether to afford
6-(methoxymethyl)pyrimidin-4-amine (0.50 g, 3.59 mmol, 48% yield)
as a pale yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 8.47 (1H, s), 6.55 (1H, s), 5.12 (2H, br. s.), 4.38 (2H, s),
3.45 (3H, s). MS (LC/MS) R.T.=0.42; [M+H].sup.+=140.20.
Step D: 4-Isothiocyanato-6-(methoxymethyl)pyrimidine
##STR00736##
[0397] To a bright orange solution of
1,1'-thiocarbonyldipyridin-2(1H)-one (0.84 g, 3.59 mmol) was in
dichloromethane at ambient temperature was added
6-(methoxymethyl)pyrimidin-4-amine (0.5 g, 3.59 mmol). The orange
solution was stirred at ambient temperature for 18 h. The solution
was purified by column chromatography (0-40% ethyl acetate/hexanes)
to afford 4-isothiocyanato-6-(methoxymethyl)pyrimidine (0.32 g,
1.77 mmol, 49% yield) as a yellow solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 8.91 (1H, d, J=1.26 Hz), 7.19 (1H, d,
J=1.01 Hz), 4.52 (2H, s), 3.49 (3H, s). MS (LC/MS) R.T.=1.39;
[M+H].sup.+=182.10.
Step E:
(R)--N-(6-(Methoxymethyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine
##STR00737##
[0399] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.39
g, 1.71 mmol) in dimethylformamide was added cesium carbonate (1.39
g, 4.28 mmol) and 4-isothiocyanato-6-(methoxymethyl)pyrimidine
(0.31 g, 1.71 mmol). The suspension was stirred at ambient
temperature for 15 min. To the reaction mixture was added
N,N'-diisopropylcarbodiimide (0.80 mL, 5.13 mmol) and the mixture
was stirred overnight then concentrated. The residue was purified
by column chromatography (5-25% 9:1 methanol/ammonium hydroxide in
ethyl acetate) to afford
(R)--N-(6-(methoxymethyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine (0.18 g, 0.58 mmol, 34% yield) as a yellow
solid. .sup.1H NMR (400 MHz, MeOD-d.sub.4) .delta. ppm 8.65 (1H, d,
J=1.01 Hz), 6.92 (1H, br. s.), 4.40 (3H, s), 4.03 (1H, d, J=10.32
Hz), 3.72 (1H, d, J=10.32 Hz), 3.44 (2H, s), 3.18-3.26 (1H, m),
3.06-3.13 (1H, m), 2.69-2.96 (4H, m), 1.94-2.19 (2H, m), 1.45-1.86
(3H, m). MS (LC/MS) R.T.=0.76; [M+H].sup.+=304.30.
Example 219
(R)--N-(5-(Cyclopentyloxy)pyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00738##
[0400] Step A: 2-Chloro-5-(cyclopentyloxy)pyrimidine
##STR00739##
[0402] A mixture of 2-chloropyrimidin-5-ol (1 g, 7.66 mmol),
chlorocyclopentane (2.39 mL, 22.98 mmol) and potassium carbonate
(3.18 g, 22.98 mmol) in N,N-dimethylformamide were heated at
65.degree. C. for 16 h at ambient temperature. Water was added and
the mixture was extracted with ethyl acetate. The organic layer was
washed with water and brine, dried over sodium sulfate and
concentrated in vacuo. The residue was purified by column
chromatography (0-25% ethyl acetate/hexanes) to afford
2-chloro-5-(cyclopentyloxy)pyrimidine (831 mg, 4.18 mmol, 54.6%
yield) as a white solid. MS (LC/MS) R.T.=2.32;
[M+H].sup.+=199.23.
Step B: 5-(Cyclopentyloxy)pyrimidin-2-amine
##STR00740##
[0404] 5-(Cyclopentyloxy)pyrimidin-2-amine was prepared from
2-chloro-5-(cyclopentyloxy)pyrimidine by following the general
procedures of Example 218, Step C. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 7.99 (2H, s), 6.18 (2H, s), 4.54-4.75
(1H, m), 1.30-1.91 (8H, m). MS (LC/MS) R.T.=1.47;
[M+H].sup.+=180.24.
Step C:
(R)--N-(5-(Cyclopentyloxy)pyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR00741##
[0406]
(R)--N-(5-(Cyclopentyloxy)pyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine was prepared from
5-(cyclopentyloxy)pyrimidin-2-amine by following the general
procedures of Example 23, Steps A-B. .sup.1H NMR (400 MHz,
MeOD-d.sub.4) .delta. ppm 8.24 (2H, s), 4.76-4.86 (1H, m), 3.98
(1H, d, J=10.07 Hz), 3.69 (1H, d, J=10.07 Hz), 3.33 (1H, d), 3.20
(1H, d), 2.77-3.08 (4H, m), 2.04-2.26 (2H, m), 1.49-2.03 (11H, m).
MS (LC/MS) R.T.=1.56; [M+H].sup.+=344.32.
[0407] The compounds in Table 5 were synthesized according to the
method of Example 218 using the appropriate commercially available
chlorides as in Example 218, Step C.
TABLE-US-00006 TABLE 4 ##STR00742## LCMS Example LCMS Ion Number
R.sub.1 RT (min) [M + H].sup.+ .sup.1H NMR 220 ##STR00743## 0.99
310.30 .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.82 (1 H, br.
s.), 8.62 (1 H, s), 7.95 (1 H, dd, J = 8.18, 1.13 Hz), 7.72 (1 H,
dd, J = 8.18, 1.13 Hz), 7.58-7.64 (1 H, m, J = 7.62, 7.62, 7.05,
1.39 Hz), 7.51 (1 H, ddd, J = 7.68, 7.05, 1.26 Hz), 4.05 (1 H, d, J
= 9.32 Hz), 3.70 (1 H, d, J = 9.57 Hz), 3.39 (1 H, dd, J = 14.86,
1.51 Hz), 2.72-3.06 (5 H, m), 2.12-2.26(2 H, m), 1.69- 1.80 (1 H,
m), 1.45-1.64 (2 H, m). M.P. 212-5.degree. C. 221 ##STR00744## 1.32
180.09 .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 8.31 (1 H,
s), 7.80 (1 H, d, J = 2.52 Hz), 7.55 (1 H, dd, J = 8.81, 2.52 Hz),
7.50 (1 H, d, J = 9.06 Hz), 7.13 (2 H, s) 222 ##STR00745## 0.90
316.30 .sup.1H NMR (400 MHz, MeOD-d.sub.4) .delta. ppm 8.60 (s, 1
H), 7.60 (d, J = 6.04 Hz, 1 H), 7.52 (s, 1 H), 4.04-4.17 (m, 1 H),
3.73-3.87 (m, 1 H), 3.23-3.26 (m, 1 H), 3.07-3.19 (m, 1 H),
2.88-3.01 (m, 2 H), 2.75-2.88 (m, 2 H), 2.08-2.27 (m, 2 H),
1.56-1.86 (m, 3 H) 223 ##STR00746## 2.28 358.20 .sup.1H NMR (400
MHz, MeOD-d.sub.4) .delta. ppm 8.52 (1 H, s), 7.30 (1 H, d, J =
1.01 Hz), 4.03-4.09 (1 H, m), 3.76 (1 H, d, J = 10.07 Hz),
3.16-3.27 (2 H, m), 3.06-3.14 (1 H, m), 2.68-3.01 (4 H, m),
2.01-2.23 (2 H, m), 1.54-1.82 (3 H, m), 1.37 (6 H, d, J = 6.80 Hz)
224 ##STR00747## 0.86 302.24 .sup.1H NMR (400 MHz, MeOD-d.sub.4)
.delta. ppm 8.63 (1 H, s), 6.73 (1 H, br. s.), 4.02 (1 H, d, J =
10.32 Hz), 3.72 (1 H, d, J = 10.32 Hz), 3.17-3.25 (1 H, m),
3.01-3.13 (1 H, m), 2.68-2.97 (5 H, m), 1.98-2.16 (2 H, m),
1.51-1.82 (3 H, m), 1.24 (6 H, d, J = 7.05 Hz) 225 ##STR00748##
0.25 274.19 .sup.1H NMR (400 MHz, MeOD-d.sub.4) .delta. ppm 8.60 (1
H, s), 6.71 (1 H, br. s.), 4.02 (1 H, d, J = 10.32 Hz), 3.71 (1 H,
d, J = 10.32 Hz), 3.17-3.24 (1 H, m), 3.04-3.13 (1 H, m), 2.65-3.02
(4 H, m), 2.37 (3 H, s), 1.99-2.20 (2 H, m), 1.32-1.88 (3 H, m) 226
##STR00749## 0.37 274.26 .sup.1H NMR (400 MHz, MeOD-d.sub.4)
.delta. ppm 8.38 (2 H, s), 3.97 (1 H, d, J = 10.07 Hz), 3.66 (1 H,
d, J = 10.07 Hz), 3.16-3.25 (1 H, m), 3.02-3.13 (1 H, m), 2.92(2 H,
t, J = 7.55 Hz), 2.73-2.86 (2 H, m), 2.21 (3 H, s), 2.03-2.16 (2 H,
m), 1.45-1.86 (3 H, m) 227 ##STR00750## 0.96 288.31 .sup.1H NMR
(400 MHz, MeOD-d.sub.4) .delta. ppm 8.41 (2 H, s), 3.98 (1 H, d, J
= 10.07 Hz), 3.67 (1 H, d, J = 10.07 Hz), 3.17-3.25 (1 H, m),
3.02-3.13 (1 H, m), 2.92 (2 H, t, J = 7.43 Hz), 2.72-2.87 (2 H, m),
2.57 (2 H, q, J = 7.55 Hz), 2.01-2.23 (2 H, m), 1.48-1.84 (3 H, m),
1.22 (3 H, t, J = 7.68 Hz)
Example 228
(R)--N-(6-(2,2,2-Trifluoroethoxy)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine
##STR00751##
[0408] Step A: 4-Chloro-6-(2,2,2-trifluoroethoxy)pyrimidine
##STR00752##
[0410] A solution of 2,2,2-trifluoroethanol (2.61 g, 26.10 mmol) in
tetrahydrofuran (12 ml) was added dropwise to a suspension of
sodium hydride (1.31 g, 32.60 mmol) in tetrahydrofuran (48 ml) at
0.degree. C. The mixture was stirred at 0.degree. C. for 30 min and
a solution of 4,6-dichloropyrimidine (3.6 g, 24.16 mmol) in
tetrahydrofuran (12 ml) was added at 0.degree. C. The reaction
mixture was stirred at ambient temperature for 3 h and poured into
sat. aqueous ammonium chloride and extracted with ethyl acetate.
The ethyl acetate extract was washed with water, dried over
magnesium sulfate and concentrated in vacuo. The orange residue was
purified by column chromatography (10-40% ethyl acetate/hexanes) to
afford 4-chloro-6-(2,2,2-trifluoroethoxy)pyrimidine (2.0 g, 2.41
mmol, 38.9% yield) as a pale yellow oil. MS (LC/MS) R.T.=2.78;
[M+H].sup.+=213.12.
Step B: 6-(2,2,2-Trifluoroethoxy)pyrimidin-4-amine
##STR00753##
[0412] 6-(2,2,2-trifluoroethoxy)pyrimidin-4-amine was prepared from
4-chloro-6-(2,2,2-trifluoroethoxy)pyrimidine by following the
general procedure of Example 218, Step C. MS (LC/MS) R.T.=1.16;
[M+H].sup.+=194.07.
Step C:
(R)--N-(6-(2,2,2-Trifluoroethoxy)pyrimidin-4-yl)-4H-1'-azaspiro[ox-
azole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00754##
[0414]
(R)--N-(6-(2,2,2-trifluoroethoxy)pyrimidin-4-yl)-4H-1'-azaspiro[oxa-
zole-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared from
6-(2,2,2-trifluoroethoxy)pyrimidin-4-amine by following the general
procedures of Example 23, Steps A-B. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 9.30 (1H, br. s.), 8.39 (1H, s), 6.38 (1H,
br. s.), 4.59-4.84 (2H, m), 3.94 (1H, d, J=9.32 Hz), 3.59 (1H, d,
J=9.57 Hz), 3.33 (1H, d, J=16.62 Hz), 2.61-3.01 (5H, m), 1.97-2.25
(2H, m), 1.37-1.81 (3H, m). MS (LC/MS) R.T.=1.42;
[M+H].sup.+=358.33.
Example 229
[0415]
(R)--N-(5-Bromo-4-isopropylpyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR00755##
Step A: 4-Isopropylpyrimidin-2-amine
##STR00756##
[0417] 4-Isopropylpyrimidin-2-amine was prepared from
2-chloro-4-isopropylpyrimidine by following the general procedure
for Example 218, Step C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) d ppm
8.10 (1H, d, J=5.04 Hz), 6.45 (3H, d, J=5.04 Hz), 2.59-2.80 (1H,
m), 1.15 (6H, d, J=7.05 Hz). MS (LC/MS) R.T.=0.76;
[M+H].sup.+=138.12.
Step B: 5-Bromo-4-isopropylpyrimidin-2-amine
##STR00757##
[0419] N-Bromosuccinimide (0.5 g, 2.8 mmol) was added to a solution
of 4-isopropylpyrimidin-2-amine (0.39 g, 2.81 mmol) in chloroform.
The resultant yellow solution was stirred at ambient temperature
for 1 h and concentrated in vacuo. The residue was purified by
column chromatography (3-10% 9:1 methanol:ammonium hydroxide in
chloroform) to afford 5-bromo-4-isopropylpyrimidin-2-amine (0.69 g,
3.18 mmol, 113%) as a pale yellow solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 8.21 (1H, s), 6.75 (2H, s), 3.10-3.23
(1H, m), 1.14 (6H, d, J=6.80 Hz). MS (LC/MS) R.T.=2.58;
[M].sup.+=216.09.
Step C:
(R)--N-(5-Bromo-4-isopropylpyrimidin-2-yl)-4H-1'-azaspiro[oxazole--
5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00758##
[0421]
(R)--N-(5-Bromo-4-isopropylpyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine was prepared from
5-bromo-4-isopropylpyrimidin-2-amine by following the general
procedures of Example 23, Steps A-B. .sup.1H NMR (400 MHz,
MeOD-d.sub.4) .delta. ppm 8.49 (1H, s), 4.02 (1H, d, J=10.07 Hz),
3.72 (1H, d, J=10.07 Hz), 3.34-3.44 (1H, m), 3.23 (1H, s),
3.06-3.15 (1H, m), 2.95 (2H, t, J=7.55 Hz), 2.75-2.89 (2H, m),
2.00-2.21 (2H, m), 1.52-1.83 (3H, m), 1.24 (6 H, d, J=6.80 Hz). MS
(LC/MS) R.T.=1.84; [M+H].sup.+=382.24.
Example 230
(R)--N-(5-Bromo-4-(pyridin-3-yl)pyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine
##STR00759##
[0423]
(R)--N-(5-Bromo-4-(pyridin-3-yl)pyrimidin-2-yl)-4H-1'-azaspiro[oxaz-
ole-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared from
4-(pyridin-3-yl)pyrimidin-2-amine by following the general
procedures of Example 229, Steps B-C. .sup.1H NMR (400 MHz,
MeOD-d.sub.4) .delta. ppm 8.92 (1H, d, J=1.51 Hz), 8.73 (1H, s),
8.64 (1H, dd, J=4.91, 1.64 Hz), 8.23 (1H, dt, J=8.06, 1.89 Hz),
7.56 (1H, dd, J=7.93, 4.91 Hz), 4.00 (1H, d, J=10.07 Hz), 3.69 (1H,
d, J=10.07 Hz), 3.22 (1H, d), 3.07 (1H, d), 2.60-2.99 (4H, m),
2.00-2.21 (2H, m), 1.50-1.83 (3H, m). MS (LC/MS) R.T.=0.76;
[M+H].sup.+=416.30.
Example 231
(R)--N-(6-(Cyclopentyloxy)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00760##
[0424] Step A: 4-Chloro-6-(cyclopentyloxy)pyrimidine
##STR00761##
[0426] A solution of cyclopentanol (2.25 g, 26.1 mmol) in
tetrahydrofuran (12 ml) was added dropwise to a suspension of
sodium hydride (1.31 g, 32.6 mmol) in tetrahydrofuran (48 ml) at
0.degree. C. The mixture was stirred at 0.degree. C. for 30 min and
a solution of 4,6-dichloropyrimidine (3.6 g, 24.16 mmol) in
tetrahydrofuran (12 ml) was added at 0.degree. C. The reaction
mixture was stirred at ambient temperature for 3 h and poured into
sat. aqueous ammonium chloride and extracted with ethyl acetate.
The organic layers were washed with water, dried over magnesium
sulfate and concentrated in vacuo. The orange residue was purified
by column chromatography (10-40% ethyl acetate/hexanes). To afford
4-chloro-6-(cyclopentyloxy)pyrimidine as a pale yellow oil. This
material was used directly for the next reaction.
Step B: 6-(Cyclopentyloxy)pyrimidin-4-amine
##STR00762##
[0428] 6-(Cyclopentyloxy)pyrimidin-4-amine was prepared from
4-chloro-6-(cyclopentyloxy)pyrimidine by following the general
procedure of Example 218, Step C. MS (LC/MS) R.T.=1.64;
[M+H].sup.+=180.22.
Step C:
(R)--N-(6-(Cyclopentyloxy)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR00763##
[0430]
(R)--N-(6-(cyclopentyloxy)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine was prepared from
6-(cyclopentyloxy)pyrimidin-4-amine by following the general
procedures of Example 23, Steps A-B. .sup.1HNMR (400 MHz,
MeOD-d.sub.4) .delta. ppm 8.37 (1H, s), 6.16 (1H, br. s.), 5.27
(1H, br. s.), 3.98 (1H, d, J=10.32 Hz), 3.67 (1H, d, J=10.32 Hz),
3.15-3.24 (1H, m), 3.02-3.12 (1 H, m), 2.71-2.97 (4H, m), 2.01-2.14
(2H, m), 1.87-2.00 (3H, m), 1.49-1.84 (8 H, m). MS (LC/MS)
R.T.=1.96; [M+H].sup.+=344.34.
Example 232
(R)--N-(6-Isopropoxypyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.-
2.2]octan]-2-amine
##STR00764##
[0432]
(R)--N-(6-Isopropoxypyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine was prepared from 4,6-dichloropyrimidine
by following the general procedures of Example 231, Steps A-C.
.sup.1H NMR (400 MHz, MeOD-d.sub.4) .delta. ppm 8.37 (1H, s), 6.13
(1H, br. s.), 5.09-5.30 (1H, m), 3.98 (1H, d, J=10.32 Hz), 3.67
(1H, d, J=10.32 Hz), 3.13-3.24 (1H, m), 3.01-3.09 (1H, m),
2.68-2.98 (4H, m), 1.98-2.17 (2H, m), 1.49-1.83 (3H, m), 1.30 (6H,
d, J=6.04 Hz). MS (LC/MS) R.T.=1.36; [M+H].sup.+=318.24.
Example 233
(R)--N-(6-(2,2-Difluoroethoxy)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine
##STR00765##
[0434]
(R)--N-(6-(2,2-Difluoroethoxy)pyrimidin-4-yl)-4H-1'-azaspiro[oxazol-
e-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared from
4,6-dichloropyrimidine by following the general procedures of
Example 231, Steps A-C. M.P. 83-8.degree. C. 1H NMR (400 MHz,
MeOD-d.sub.4) .delta. ppm 8.42 (1H, s), 5.93-6.37 (2H, m), 4.52
(2H, td, J=13.98, 3.78 Hz), 3.99 (1H, d, J=10.32 Hz), 3.68 (1H, d,
J=10.32 Hz), 3.19 (1H, d), 3.07 (1H, d), 2.67-2.97 (4H, m),
1.99-2.19 (2H, m), 1.51-1.82 (3H, m). MS (LC/MS) R.T.=0.99;
[M+H].sup.+=340.26.
Example 234
(R)--N-(Pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-a-
mine
##STR00766##
[0436]
(R)--N-(Pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oct-
an]-2-amine was prepared from
(R)--N-(6-bromopyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oc-
tan]-2-amine (from Example 155) according to the general procedure
of Example 19, Step C. .sup.1H NMR (500 MHz, MeOD-d.sub.4) .delta.
ppm 8.26 (d, J=4.88 Hz, 1H), 7.58-7.74 (m, 1H), 6.85-7.02 (m, 2H),
4.00 (d, J=10.07 Hz, 1 H), 3.70 (d, J=10.07 Hz, 1H), 3.26-3.35 (m,
1H), 3.14-3.21 (m, 1H), 3.02 (d, J=8.24 Hz, 2H), 2.84-2.97 (m, 2H),
2.11-2.25 (m, 2H), 1.58-1.92 (m, 3H). MS (LC/MS) R.T.=0.30;
[M+H].sup.+=259.16.
Example 235
(R)--N-(Pyridin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-a-
mine
##STR00767##
[0438]
(R)--N-(Pyridin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oct-
an]-2-amine was prepared from
(R)--N-(2-bromopyridin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oc-
tan]-2-amine (from Example 154) according to the procedure of
Example 19, Step C. .sup.1H NMR (500 MHz, MeOD-d.sub.4) .delta. ppm
8.31 (d, J=6.41 Hz, 2 H), 7.39 (d, J=3.97 Hz, 2H), 4.01 (d, J=12.21
Hz, 1H), 3.70 (d, J=11.90 Hz, 1H), 3.37 (s, 1H), 3.29 (s, 1H), 3.18
(d, J=1.83 Hz, 1H), 3.15 (d, J=2.14 Hz, 1H), 2.86-3.09 (m, 3H),
2.02-2.20 (m, 1H), 1.59-1.88 (m, 3H). MS (LC/MS) R.T.=0.22;
[M+H].sup.+=259.16.
Example 236
(R)--N-(5-(Benzyloxy)thiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR00768##
[0439] Step A: 5-(Benzyloxy)thiazolo[5,4-b]pyridin-2-amine
##STR00769##
[0441] Potassium thiocyanate (12.42 g, 128 mmol) was suspended in
acetic acid (45.0 mL) and cooled to 0.degree. C.
6-(Benzyloxy)pyridin-3-amine (3.2 g, 15.98 mmol), prepared
according to WO2006/044707 was added. Bromine (2.55 mL, 49.5 mmol)
in acetic acid (15 mL) was added dropwise over 30 minutes during
which time the reaction mixture became very thick. It was allowed
to warm to room temperature slowly and stirred overnight.
[0442] Water (20 ml) was added and the reaction mixture was heated
to 90.degree. C. and filtered hot. The filtrate was saved and the
filter cake returned to the reaction flask, to which was added an
additional 40 ml HOAc. The mixture was again heated to 90.degree.
C. and filtered hot. The combined filtrates were cooled on ice bath
and NH.sub.4OH was added dropwise until pH>8. A yellow
precipitate formed which was collected by filtration. The solids
were dried in vacuo for 1 h to provide
5-(benzyloxy)thiazolo[5,4-b]pyridin-2-amine (1.95 g, 7.58 mmol,
47.4% yield), which was used without further purification. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. ppm 7.70 (d, J=8.78 Hz, 1H) 7.48
(d, J=7.28 Hz, 2H) 7.39 (t, J=7.28 Hz, 2H) 7.30-7.36 (m, 1 H) 6.78
(d, J=8.78 Hz, 1H) 5.39 (s, 2H) 5.14 (br. s., 2H).
Step B: Dimethyl
5-(benzyloxy)thiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate
##STR00770##
[0444] To a suspension of
5-(benzyloxy)thiazolo[5,4-b]pyridin-2-amine (800 mg, 3.11 mmol) in
DMF (3.1 mL) was added 20.0M sodium hydroxide (0.3 mL, 6.22 mmol).
The mixture was allowed to stir 10 min at room temperature at which
time carbon disulfide was added (0.47 mL, 7.77 mmol) and the
mixture was stirred for 10 minutes. An additional portion of 20.0M
sodium hydroxide (0.3 mL, 6.22 mmol) was added and the mixture was
again stirred for 10 minutes. Finally, iodomethane (0.47 mL, 7.46
mmol) was added dropwise. The mixture was stirred for 1 hour, at
which time it was poured into water and extracted with EtOAc
(3.times.). The combined organics were washed with brine, dried
over sodium sulfate, filtered and concentrated in vacuo. The crude
mixture was purified by silica gel chromatography (2-20%
EtOAc/CHCl.sub.3) to provide dimethyl
5-(benzyloxy)thiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate (1.02
g, 91% yield) as a yellow crystalline solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 8.02 (d, J=8.78 Hz, 1H) 7.51 (d, J=7.28 Hz,
2H) 7.38-7.45 (m, 2H) 7.32-7.38 (m, 1H) 6.89 (d, J=8.53 Hz, 1H)
5.46 (s, 2H) 2.65 (s, 6H).
Step C:
(R)--N-(5-(benzyloxy)thiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[o-
xazole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00771##
[0446] A mixture of dimethyl
5-(benzyloxy)thiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate (500
mg, 1.38 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(317 mg, 1.38 mmol) and cesium carbonate (1.0 g, 3.07 mmol) in DMF
(7 mL) was heated to 100.degree. C. for 2 hours. The reaction
mixture was cooled to ambient temperature, poured into water and
extracted with chloroform (4.times.). The combined organics were
washed with brine, dried over sodium sulfate, filtered and
concentrated in vacuo. The mixture was purified by silica gel
chromatography (2-20% [9:1 methanol:ammonium hydroxide]-chloroform)
to afford
(R)--N-(5-(benzyloxy)thiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole--
5,3'-bicyclo[2.2.2]octan]-2-amine (390 mg, 67% yield). .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 9.18 (br. s., 1H) 7.76 (d, J=8.85
Hz, 1H) 7.50 (d, J=7.32 Hz, 2H) 7.41 (t, J=7.32 Hz, 2H) 7.32-7.37
(m, 1H) 5.42 (s, 2H) 4.02 (d, J=9.46 Hz, 1H) 3.68 (d, J=9.46 Hz,
1H) 3.37-3.44 (m, 1H) 2.75-3.06 (m, 5H) 2.13-2.25 (m, 2H) 1.73-1.82
(m, 1H) 1.49-1.70 (m, 3H). MS (LC/MS) R.T.=1.69;
[M+H].sup.+=421.98.
Example 237
(R)-2-(4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)thiazol-
o[5,4-b]pyridin-5(4H)-one
##STR00772##
[0448]
(R)--N-(5-(benzyloxy)thiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[ox-
azole-5,3'-bicyclo[2.2.2]octan]-2-amine (390 mg, 0.925 mmol) was
dissolved in TFA and allowed to react for 4 hours at ambient
temperature, at which time LCMS and TLC showed the starting
material to be mostly consumed. The TFA was removed in vacuo and
the crude mixture was purified by preparative HPLC. The combined
product fractions were concentrated in vacuo and triturated with
ether to afford
(R)-2-(4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)thiazo-
lo[5,4-b]pyridin-5(4H)-one, TFA (164 mg, 0.368 mmol, 39.8% yield).
M.P. 245 (dec). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
9.99 (br. s., 1H) 9.05 (br. s., 1H) 7.81 (d, J=8.53 Hz, 1H) 6.65
(d, J=8.78 Hz, 1H) 3.96 (d, J=10.29 Hz, 1H) 3.82 (d, J=10.54 Hz,
1H) 3.63-3.78 (m, 2H) 3.36-3.47 (m, 1H) 3.16-3.34 (m, 3H) 2.43 (br.
s., 1H) 2.16 (br. s., 1H) 1.76-2.07 (m, 3H). MS (LC/MS) R.T.=0.50;
[M+H].sup.+=332.15.
Example 238
(R)--N-(6-(3-Methoxyphenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00773##
[0449] Step A: 6-(3-Methoxyphenyl)pyrimidin-4-amine
##STR00774##
[0451] A mixture of 6-chloropyrimidin-4-amine (0.324 g, 2.5 mmol),
3-methoxyphenylboronic acid (0.475 g, 3.13 mmol), Na.sub.2CO.sub.3
(0.795 g, 7.50 mmol) and bis(triphenylphosphine)palladium(II)
chloride (0.035 g, 0.050 mmol) was suspended in DME/EtOH/water
(15:2:3 mL), heated in the microwave synthesizer at 125.degree. C.
for 20 min and concentrated. The residue was purified by silica gel
chromatography (10-60% ethyl acetate-hexanes) to afford
6-(3-methoxyphenyl)pyrimidin-4-amine (0.35 g, 1.74 mmol, 70% yield)
as an off-white solid. LCMS R.T.=1.28; [M+H].sup.+=201.98.
Step B: 4-Isothiocyanato-6-(3-methoxyphenyl)pyrimidine
##STR00775##
[0453] To a solution of 1,1'-thiocarbonyldipyridin-2(1H)-one (0.970
g, 4.17 mmol) in dichloromethane at room temperature was added
6-(3-methoxyphenyl)pyrimidin-4-amine (0.7 g, 3.48 mmol). The
reaction was stirred at room temperature for 18 hours. The LC/MS
showed the desired product peak as a major peak. The deep orange
solution was concentrated and the remaining residue was filtered.
The filtrate was purified by silica gel chromatography (0-10% ethyl
acetate-hexanes) to afford
4-isothiocyanato-6-(3-methoxyphenyl)pyrimidine (0.39 g, 4.31 mmol,
46% yield) as a yellow oil. LCMS R.T.=2.91; [M+H].sup.+=244.03.
Step C:
R)--N-(6-(3-Methoxyphenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR00776##
[0455] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.363 g, 1.583 mmol) in N,N-dimethylformamide (20 mL) was added
Cs.sub.2CO.sub.3 (1.289 g, 3.96 mmol) and
4-isothiocyanato-6-(3-methoxyphenyl)pyrimidine. The suspension was
stirred at room temperature for 30 minutes.
N,N'-diisopropylcarbodiimide (0.740 mL, 4.75 mmol) was then added
and the mixture was continued to stir at room temperature for 18
hours. The mixture was concentrated and purified by silica gel
chromatography (5-25% 9:1 methanol:ammonium hydroxide-ethyl
acetate) to afford
(R)--N-(6-methoxypyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (0.294 g, 0.788 mmol, 50% yield) as a pale yellow
solid. M.P 80-5.degree. C. .sup.1H NMR (400 MHz, MeOD) .delta. ppm
8.77 (1H, s), 7.47-7.58 (2H, m), 7.39 (1H, t), 7.20 (1H, br. s.),
7.04 (1H, dd), 4.05 (1H, d), 3.85 (3H, s), 3.74 (1H, d), 3.23 (1H,
d), 3.10 (1 H, d), 2.71-3.00 (4H, m), 2.03-2.22 (2H, m), 1.53-1.85
(3H, m). MS (LC/MS) R.T.=1.58; [M+H].sup.+=366.15.
Example 239
(R)--N-(Isoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-
-2-amine
##STR00777##
[0456] Step A: 3-Isothiocyanatoisoquinoline
##STR00778##
[0458] To a solution of 1,1'-thiocarbonyldipyridin-2(1H)-one (0.805
g, 3.47 mmol) in dichloromethane at room temperature was added
isoquinolin-3-amine (0.5 g, 3.47 mmol). The reaction was stirred at
room temperature for 18 hours. The LC/MS showed the desired product
peak a major peak. The deep orange solution was concentrated and
filtered. The filtrate was purified by silica gel chromatography
(0-40% ethyl acetate-hexanes) to afford
4-isothiocyanato-6-(3-methoxyphenyl)pyrimidine (0.55 g, 2.96 mmol,
85% yield) a white solid. LCMS R.T.=2.47; [M+H].sup.+=187.23.
Step B:
(R)--N-(Isoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00779##
[0460] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.2
g, 0.873 mmol) in N,N-dimethylformamide (20 mL) was added
Cs.sub.2CO.sub.3 (0.711 g, 2.182 mmol) and
3-isothiocyanatoisoquinoline (0.163 g, 0.873 mmol). The suspension
was stirred at room temperature for 30 minutes.
N,N'-Diisopropylcarbodiimide (0.408 mL, 2.62 mmol) was then added
and the mixture was stirred at room temperature for 18 hours. The
mixture was concentrated and purified by silica gel chromatography
(5-25% [9:1 methanol:ammonium hydroxide]-ethyl acetate) to afford
(R)--N-(isoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan-
]-2-amine (0.16 g, 0.508 mmol, 58% yield) as an off-white solid.
M.P. 196-200.degree. C. .sup.1H NMR (400 MHz, MeOD) .delta. ppm
9.00 (1H, s), 7.92 (1H, d), 7.71 (1H, d), 7.59 (1H, t), 7.20-7.45
(2H, m), 3.96 (1H, d), 3.65 (1H, d), 3.22 (1H, d), 3.08 (1H, d),
2.66-3.00 (4H, m), 2.05-2.23 (2H, m), 1.50-1.86 (3H, m). R.T.=1.37;
[M+H].sup.+=309.31.
Example 240
(R)--N-(6-Phenoxypyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00780##
[0461] Step A: 6-Phenoxypyrimidin-4-amine
##STR00781##
[0463] 6-Chloropyrimidin-4-amine (3.00 g, 23.14 mmol) was added to
a solution of sodium (0.197 g, 8.57 mmol) in phenol (11.29 g, 120
mmol) at 55.degree. C. The mixture was heated at 140.degree. C. for
2 h, then held at room temperature for 20 h. The reaction mixture
was poured into 32% aqueous NaOH on ice/water keeping the mixture
temperature below 20.degree. C. The mixture was extracted with
chloroform and the organic extract dried over calcium chloride and
concentrated. The residue was purified by silica gel chromatography
(2-20% ethyl acetate in hexanes) to afford
6-phenoxypyrimidin-4-amine (0.6 g, 3.21 mmol, 75% yield) as a white
solid. LCMS R.T.=1.37; [M+H].sup.+=197.95.
Step B: 4-Isothiocyanato-6-phenoxypyrimidine
##STR00782##
[0465] A mixture of 6-phenoxypyrimidin-4-amine (0.288 g, 1.538
mmol) and 1,1'-thiocarbonyldipyridin-2(1H)-one (0.357 g, 1.538
mmol) in DCM was stirred at rt for 18 h. The pale orange mixture
was purified by silica gel chromatography (5-35% ethyl
acetate-hexanes) to afford 4-isothiocyanato-6-phenoxypyrimidine
(0.55 g, 2.96 mmol, 85% yield) a yellow oil. LCMS R.T.=2.78;
[M+H].sup.+=229.94.
Step C:
(R)--N-(6-Phenoxypyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00783##
[0467] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.170 g, 0.742 mmol) in N,N-dimethylformamide (20 mL) was added
Cs.sub.2CO.sub.3 (0.604 g, 1.854 mmol) and
4-isothiocyanato-6-phenoxypyrimidine (0.17 g, 0.742 mmol). The
suspension was stirred at room temperature for 30 minutes.
N,N'-diisopropylcarbodiimide (0.347 mL, 2.225 mmol) was then added
and the mixture was continued to stir at room temperature for 18
hours. The mixture was concentrated and purified by silica gel
chromatography (5-25% 9:1 methanol:ammonium hydroxide-ethyl
acetate) to afford
(R)--N-(6-phenoxypyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine (0.21 g, 0.72 mmol, 48.2% yield) as a pale
yellow solid. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.43 (1H, s),
7.47 (2H, t), 7.30 (1H, t), 7.16 (2H, d), 6.21 (1H, br. s.), 4.03
(1H, d), 3.72 (1 H, d), 3.22 (1H, d), 3.11 (1H, d), 2.73-2.99 (4H,
m), 2.00-2.18 (2H, m), 1.54-1.88 (3H, m). LCMS R.T.=1.46;
[M+H].sup.+=352.19.
Example 241
(R)--N-(7-methoxyquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00784##
[0468] Step A:
N-(2,4-Dimethoxybenzyl)-7-methoxyquinoxalin-2-amine
##STR00785##
[0470] 2-Chloro-7-methoxyquinoxaline (0.51 g, 2.62 mmol), prepared
according to J. Chem. Soc. Perk Trans. 1, 2001, 978-984, and
(2,4-dimethoxyphenyl)methanamine (1.181 mL, 7.86 mmol) were
microwaved in DMSO (2.5 mL) for 30 min at 150.degree. C. This was
diluted into 150 mL EtOAc and extracted three times with 100 mL
brine. The crude product was purified by flash chromatography on a
90 g silica gel cartridge with 20 to 80% EtOAc in hexane, 50 min,
at 40 mL/min to afford
N-(2,4-dimethoxybenzyl)-7-methoxyquinoxalin-2-amine (795 mg, 2.443
mmol, 93% yield).
[0471] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.00 (1H, s),
7.70 (1H, d, J=8.81 Hz), 7.29 (1 H, d, J=8.31 Hz), 7.05 (1H, d,
J=2.77 Hz), 6.97 (1H, dd, J=9.06, 2.77 Hz), 6.47 (1H, d, J=2.27
Hz), 6.43 (1H, dd, J=8.18, 2.39 Hz), 5.22 (1H, t, J=5.29 Hz), 4.63
(2 H, d, J=5.54 Hz), 3.91 (3H, s), 3.83 (3H, s), 3.78 (3H, s)
[0472] LCMS: RT=1.91 min, MH+=326.15.
Step B: 7-Methoxyquinoxalin-2-amine 2,2,2-trifluoroacetate
##STR00786##
[0474] N-(2,4-Dimethoxybenzyl)-7-methoxyquinoxalin-2-amine (0.79 g,
2.428 mmol) was stirred in TFA (10 mL, 130 mmol)/CH.sub.2Cl.sub.2
(10 mL) at room temperature for 30 min. Solvents were removed on
the rotary evaporator. Saturated aqueous NaHCO.sub.3 (200 mL) was
added to the red residue, which precipitated a yellow solid. The
mixture was extracted extensively with DCM. The organic layer was
concentrated and dried under vacuum to yield
7-methoxyquinoxalin-2-amine 2,2,2-trifluoroacetate (0.70 g, 2.4
mmol, 99% yield).
[0475] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 8.10 (1H,
s), 7.63 (1H, d, J=9.07 Hz), 6.95 (1H, dd, J=9.06, 2.77 Hz), 6.89
(1H, d, J=2.77 Hz), 6.85 (2H, br. s.), 3.84 (3H, s) LCMS: RT=1.04
min, MH+=176.14.
Step C: 2-Isothiocyanato-7-methoxyquinoxaline
##STR00787##
[0477] A mixture of 7-methoxyquinoxalin-2-amine
2,2,2-trifluoroacetate (578 mg, 2 mmol), triethylamine (335 .mu.L,
2.400 mmol), and 1,1'-thiocarbonyldipyridin-2(1H)-one (557 mg,
2.400 mmol) was stirred in 5 mL DCM for 24 h. The reaction was
directly eluted on a 120 g silica gel cartridge with 0 to 25% EtOAc
in hexane, 25 min, at 35 mL/min to afford
2-isothiocyanato-7-methoxyquinoxaline (84 mg, 0.387 mmol, 19%
yield).
[0478] LCMS: RT=2.49 min, MH+=218.06.
Step D:
(R)--N-(7-Methoxyquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00788##
[0480]
(R)--N-(7-Methoxyquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine was synthesized by the method of Example
23, Step B. Flash chromatography on a 120 g silica gel cartridge
with 1-4% [9:1 MeOH/NH4OH] in CHCl3, 50 min, afforded 24 mg (17%
yield).
[0481] 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.73 (1H, br. s.),
8.45 (1H, s), 7.80 (1H, d, J=9.07 Hz), 7.12 (1H, dd, J=9.06, 2.77
Hz), 7.03 (1H, d, J=2.52 Hz), 4.02 (1H, d, J=9.32 Hz), 3.90 (3H,
s), 3.67 (1H, d, J=9.32 Hz), 3.36 (1H, dd, J=14.86, 1.51 Hz),
2.69-3.06 (5H, m), 2.10-2.26 (2H, m), 1.66-1.80 (1H, m), 1.43-1.63
(2H, m) LCMS: RT=0.835 min, MH-=338.2, MH+=340.1.
Example 242
(R)--N-(6-Methylquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00789##
[0482] Step A:
N-(2,4-Dimethoxybenzyl)-6-methylquinoxalin-2-amine
##STR00790##
[0484] 2-Chloro-6-methylquinoxaline (0.51 g, 2.86 mmol), prepared
according to J. Chem. Soc. 1948, 1310-1313, and
(2,4-dimethoxyphenyl)methanamine (1.29 mL, 8.57 mmol) were
microwaved in DMSO (2.5 mL) for 30 min at 150.degree. C. This was
diluted into 150 mL EtOAc and extracted three times with 100 mL
brine. The crude product was purified by flash chromatography on a
90 g silica gel cartridge with 20 to 60% EtOAc in hexane, 50 min,
at 40 mL/min to afford
N-(2,4-dimethoxybenzyl)-6-methylquinoxalin-2-amine (848 mg, 2.74
mmol, 96% yield).
[0485] 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.12 (1H, s), 7.60
(1H, s), 7.59 (1H, d, J=5.79 Hz), 7.38 (1H, dd, J=8.56, 1.76 Hz),
7.30 (1H, d, J=8.31 Hz), 6.46 (1H, d, J=2.27 Hz), 6.42 (1H, dd,
J=8.18, 2.39 Hz), 5.20 (1H, t, J=5.41 Hz), 4.62 (2H, d, J=5.54 Hz),
3.83 (3H, s), 3.77 (3H, s), 2.46 (3H, s)
[0486] LCMS: RT=1.93 min, MH+=310.20.
Step B: 6-Methylquinoxalin-2-amine 2,2,2-trifluoroacetate
##STR00791##
[0488] N-(2,4-Dimethoxybenzyl)-6-methylquinoxalin-2-amine (0.84 g,
2.72 mmol) was stirred in TFA (10 mL, 130 mmol)/CH.sub.2Cl.sub.2
(10 mL) at room temperature for 30 min. Solvents were removed on
the rotary evaporator. Saturated aqueous Na.sub.2CO.sub.3 (200 mL)
was added to the red residue, which then precipitated a tan solid.
The mixture was extracted extensively with DCM. The organic layer
was dried over sodium sulfate, concentrated, and dried under vacuum
to afford 6-methylquinoxalin-2-amine 2,2,2-trifluoroacetate (640
mg, 2.343 mmol, 86% yield).
[0489] 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 8.24 (1H, s),
7.55 (1H, s), 7.34-7.45 (2H, m), 6.82 (2H, s), 2.41 (3H, s)
[0490] LCMS: RT=1.07 min, MH+=160.12.
Step C: 2-Isothiocyanato-6-methylquinoxaline
##STR00792##
[0492] A mixture of 6-methylquinoxalin-2-amine
2,2,2-trifluoroacetate (546 mg, 2 mmol), triethylamine (243 mg,
2.400 mmol), and 1,1'-thiocarbonyldipyridin-2(1H)-one (557 mg, 2.40
mmol) was stirred in 5 mL DCM for 4 h. The reaction was directly
eluted on a 120 g silica gel cartridge with 0 to 25% EtOAc in
hexane, 25 min, at 35 mL/min to afford
2-isothiocyanato-6-methylquinoxaline (153 mg, 0.760 mmol, 38%
yield).
[0493] LCMS: RT=2.59 min, MH+=202.04
Step D:
(R)--N-(6-Methylquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00793##
[0495]
(R)--N-(6-Methylquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine was synthesized by the method of Example 23,
Step B. Flash chromatography on a 120 g silica gel cartridge with 1
to 4% [9:1 MeOH/NH.sub.4OH] in CHCl.sub.3, 50 min, afforded 46 mg
(19% yield).
[0496] 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.75 (1H, br. s.),
8.57 (1H, s), 7.71 (1H, s), 7.60 (1H, d, J=8.56 Hz), 7.42 (1H, dd,
J=8.44, 1.89 Hz), 4.01 (1H, d, J=9.57 Hz), 3.68 (1H, d, J=9.32 Hz),
3.38 (1H, dd, J=14.86, 1.01 Hz), 2.73-3.08 (5H, m), 2.50 (3H, s),
2.16-2.26 (1H, m), 2.14 (1H, br. s.), 1.67-1.79 (1H, m), 1.45-1.65
(2H, m).
[0497] LCMS: RT=0.838 min, MH-=322.2, MH+=324.2.
Example 243
(R)--N-(7-Methoxyquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00794##
[0498] Step A:
N-(2,4-Dimethoxybenzyl)-7-methylquinoxalin-2-amine
##STR00795##
[0500] 2-Chloro-7-methylquinoxaline (0.51 g, 2.86 mmol),), prepared
according to J. Chem. Soc. 1948 1310-1313, and
(2,4-dimethoxyphenyl)methanamine (1.29 mL, 8.57 mmol) were
microwaved in DMSO (2.5 mL) for 30 min at 150.degree. C. This was
diluted into 150 mL EtOAc and extracted three times with 100 mL
brine. The crude product was purified by flash chromatography on a
90 g silica gel cartridge with 20 to 80% EtOAc in hexane, 50 min,
at 40 mL/min to afford
N-(2,4-dimethoxybenzyl)-7-methylquinoxalin-2-amine (860 mg, 2.78
mmol, 97% yield).
[0501] 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.08 (1H, s), 7.70
(1H, d, J=8.31 Hz), 7.49 (1 H, s), 7.30 (1H, d, J=8.06 Hz), 7.17
(1H, dd, J=8.31, 2.01 Hz), 6.47 (1H, d, J=2.52 Hz), 6.42 (1H, dd,
J=8.31, 2.52 Hz), 5.23 (1H, t, J=5.16 Hz), 4.63 (2H, d, J=5.79 Hz),
3.83 (3H, s), 3.78 (3H, s), 2.48 (3H, s).
[0502] LCMS: RT=1.93 min, MH+=310.20.
Step B: 7-Methylquinoxalin-2-amine 2,2,2-trifluoroacetate
##STR00796##
[0504] N-(2,4-Dimethoxybenzyl)-7-methylquinoxalin-2-amine (0.85 g,
2.75 mmol) was stirred in TFA (10 mL, 130 mmol)/CH2Cl2 (10 mL) at
room temperature for 30 min. Solvents were removed on the rotary
evaporator. Saturated aqueous NaHCO.sub.3 (200 mL) was added to the
red residue, which then precipitated a pink solid. The mixture was
extracted extensively with DCM. The organic layer was dried over
sodium sulfate, concentrated, and dried under vacuum to afford
7-methylquinoxalin-2-amine 2,2,2-trifluoroacetate (640 mg, 2.34
mmol, 85% yield).
[0505] 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 8.20 (1H, s),
7.64 (1H, d, J=8.31 Hz), 7.28 (1H, s), 7.15 (1H, dd, J=8.31, 1.76
Hz), 6.89 (2H, s), 2.42 (3H, s).
[0506] LCMS: RT=1.07 min, MH+=160.12.
Step C: 2-Isothiocyanato-6-methylquinoxaline
##STR00797##
[0508] A mixture of 7-methylquinoxalin-2-amine
2,2,2-trifluoroacetate (546 mg, 2 mmol) 78263-058-01, triethylamine
(243 mg, 2.40 mmol), and 1,1'-thiocarbonyldipyridin-2(1H)-one (557
mg, 2.40 mmol) was stirred in 5 mL DCM for 2 h. The reaction was
directly eluted on a 120 g silica gel cartridge with 0 to 25% EtOAc
in hexane, 25 min, at 35 mL/min to afford
2-isothiocyanato-7-methylquinoxaline (185 mg, 0.919 mmol, 46%
yield).
[0509] LCMS: RT=2.58 min, MH+=202.04
Step D:
(R)--N-(7-Methylquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00798##
[0511]
(R)--N-(7-Methylquinoxalin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine was synthesized by the method of Example 23,
Step B. Flash chromatography on a 120 g silica gel cartridge with 1
to 3% [9:1 MeOH/NH4OH] in CHCl3, 50 min, afforded 22 mg (7%
yield).
[0512] 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.80 (1H, br. s.),
8.53 (1H, s), 7.81 (1H, d, J=8.31 Hz), 7.50 (1H, s), 7.31 (1H, dd,
J=8.56, 1.76 Hz), 4.01 (1H, d, J=9.57 Hz), 3.66 (1H, d, J=9.32 Hz),
3.36 (1H, d, J=14.86 Hz), 2.70-3.04 (5H, m), 2.50 (3H, s),
2.15-2.24 (1H, m), 2.13 (1H, br. s.), 1.66-1.79 (1H, m), 1.44-1.63
(2H, m) LCMS: RT=8.67 min, MH-=322.6, MH+=324.1.
Example 244
(R)--N-(6-(Pyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine
##STR00799##
[0513] Step A: 6-(Pyridin-3-yl)pyrimidin-4-amine
##STR00800##
[0515] A mixture of 6-chloropyrimidin-4-amine (0.324 g, 2.5 mmol),
pyridin-3-ylboronic acid (0.384 g, 3.13 mmol), Na.sub.2CO.sub.3
(0.795 g, 7.50 mmol) and bis(triphenylphosphine)palladium(II)
chloride (0.035 g, 0.050 mmol) was suspended in a mixture of
DME/EtOH/water. The mixture was heated in the microwave synthesizer
at 125.degree. C. for 20 min and concentrated. The residue was
purified by silica gel chromatography (10-60% ethyl acetate in
hexanes, then 5-25% 9:1 methanol:ammonium hydroxide-ethyl acetate)
to afford 6-(pyridin-3-yl)pyrimidin-4-amine (0.17 g, 0.987 mmol,
40% yield) as an off-white solid. LCMS R.T.=0.31;
[M+H].sup.+=173.11.
Step B: 4-Isothiocyanato-6-(pyridin-3-yl)pyrimidine
##STR00801##
[0517] To a solution of 1,1'-thiocarbonyldipyridin-2(1H)-one (0.682
g, 2.94 mmol) in dichloromethane/N,N-dimethylformamide at room
temperature was added 6-(pyridin-3-yl)pyrimidin-4-amine (0.337 g,
1.957 mmol). The mixture was heated at 60.degree. C. for 18 hours.
LC/MS showed the desired product peak as the major peak. The deep
orange mixture was purified by silica gel chromatography (1-40%
ethyl acetate-hexanes) to afford
4-isothiocyanato-6-methoxypyrimidine (0.12 g, 0.56 mmol, 28.6%
yield) as an orange oil.
Step C:
(R)--N-(6-(Pyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine
##STR00802##
[0519] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.13
g, 0.560 mmol) in N,N-dimethylformamide (20 mL) was added
Cs.sub.2CO.sub.3 (0.46 g, 1.4 mmol) and
4-isothiocyanato-6-(pyridin-3-yl)pyrimidine (0.12 g, 0.56 mmol).
The suspension was stirred at room temperature for 30 minutes.
N,N'-Diisopropylcarbodiimide (0.26 mL, 1.7 mmol) was then added and
the mixture was stirred at room temperature for 18 hours. The
mixture was concentrated and purified by silica gel chromatography
(0-10% [9:1 methanol:ammonium hydroxide]-ethyl acetate) to afford
(S)--N-(5-chloropyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]o-
ctan]-2-amine (0.182 g, 0.613 mmol, 35% yield) as an off-white
solid. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.13-9.21 (1H, m),
8.82 (1H, d), 8.63 (1 H, dd), 8.44 (1H, dt), 7.56 (1H, dd), 7.31
(1H, s), 4.06 (1H, d), 3.76 (1H, d), 3.20-3.28 (1H, m), 3.08-3.16
(1H, m), 2.72-3.01 (4H, m), 2.00-2.24 (2H, m), 1.52-1.83 (3H, m).
LCMS R.T.=0.72; [M+H].sup.+=337.2.
Example 245
(R)--N-(2'-Methoxy-4,5'-bipyrimidin-6-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00803##
[0520] Step A: 2'-Methoxy-4,5'-bipyrimidin-6-amine
##STR00804##
[0522] 6-Chloropyrimidin-4-amine (0.35 g, 2.70 mmol),
2-methoxypyrimidin-5-ylboronic acid (0.520 g, 3.38 mmol),
Na.sub.2CO.sub.3 (0.859 g, 8.11 mmol) and
bis(triphenylphosphine)palladium(II) chloride (0.038 g, 0.054 mmol)
were suspended in a mixture of DME/EtOH/water. (15:2:3 mL). The
mixture was heated in the microwave synthesizer at 125.degree. C.
for 20 min and concentrated. The residue was purified by silica gel
chromatography (0-5% 9:1 methanol:ammonium hydroxide-ethyl acetate)
to afford 6-(pyridin-3-yl)pyrimidin-4-amine (0.28 g, 1.378 mmol,
51% yield) as an off-white solid. LCMS R.T.=0.53;
[M+H].sup.+=204.11.
Step B: 6-Isothiocyanato-2'-methoxy-4,5'-bipyrimidine
##STR00805##
[0524] To a solution of 1,1'-thiocarbonyldipyridin-2(1H)-one (0.832
g, 3.58 mmol) in dichloromethane/N,N-dimethylformamide at room
temperature was added 2'-methoxy-4,5'-bipyrimidin-6-amine (0.56 g,
2.76 mmol). The orange mixture was heated at 60.degree. C. for 18
hours. The LC/MS showed the desired product peak as the major peak.
The deep orange mixture was purified by silica gel chromatography
(0-40% ethyl acetate-hexanes) to afford
4-isothiocyanato-6-methoxypyrimidine (0.1 g, 0.408 mmol, 15% yield)
as an orange solid. LCMS R.T.=2.29; [M+H].sup.+=246.03.
Step C:
(R)--N-(2'-Methoxy-4,5'-bipyrimidin-6-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR00806##
[0526] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.093 g, 0.41 mmol) in N,N-dimethylformamide (20 mL) was added
Cs.sub.2CO.sub.3 (0.33 g, 1 mmol)) and
6-isothiocyanato-2'-methoxy-4,5'-bipyrimidine (0.1 g, 0.41 mmol).
The suspension was stirred at room temperature for 30 minutes.
N,N'-Diisopropylcarbodiimide (0.19 mL, 1.2 mmol) was then added and
the mixture was stirred at room temperature for 18 hours. The
mixture was concentrated and purified by silica gel chromatography
(5-25% [9:1 methanol:ammonium hydroxide]-ethyl acetate) to afford
(R)--N-(2'-methoxy-4,5'-bipyrimidin-6-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine (0.072 g, 0.188 mmol, 46% yield) as an
off-white solid. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.19 (2H,
s), 8.80 (1H, d), 7.24 (1H, br. s.), 4.00-4.09 (4H, m), 3.76 (1H,
d), 3.23 (1H, s), 3.08-3.15 (1H, m), 2.72-3.04 (4H, m), 1.97-2.22
(2 H, m), 1.38-1.85 (3H, m). R.T.=1.22; [M+H].sup.+=368.22.
Example 246
(R)--N-(6-(Pyridin-4-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine
##STR00807##
[0527] Step A: 6-(Pyridin-4-yl)pyrimidin-4-amine
##STR00808##
[0529] 6-Chloropyrimidin-4-amine (0.324 g, 2.5 mmol),
pyridin-4-ylboronic acid (0.384 g, 3.13 mmol), Na.sub.2CO.sub.3
(0.795 g, 7.50 mmol) and bis(triphenylphosphine)palladium(II)
chloride (0.035 g, 0.050 mmol) were suspended in a mixture of
DME/EtOH/water (15:2:3 mL). The mixture was heated in the microwave
synthesizer at 125.degree. C. for 20 min and concentrated. The
residue was purified by silica gel chromatography (5-25% [9:1
methanol:ammonium hydroxide]-ethyl acetate) to afford
6-(pyridin-3-yl)pyrimidin-4-amine (0.15 g, 0.871 mmol, 35% yield)
as an off-white solid. LCMS R.T.=0.30; [M+H].sup.+=173.11.
Step B: 4-Isothiocyanato-6-(pyridin-4-yl)pyrimidine
##STR00809##
[0531] To a solution of 1,1'-thiocarbonyldipyridin-2(1H)-one (0.601
g, 2.59 mmol) in dichloromethane/N,N-dimethylformamide at room
temperature was added 6-(pyridin-4-yl)pyrimidin-4-amine (0.297 g,
1.725 mmol). The orange mixture was heated at 60.degree. C. for 18
hours. LC/MS showed the desired product peak as the major peak. The
deep orange mixture was purified by silica gel chromatography
(0-40% ethyl acetate-hexanes) to afford
4-isothiocyanato-6-(pyridin-4-yl)pyrimidine (0.055 g, 0.257 mmol,
15% yield) as an orange solid. LCMS R.T.=1.46;
[M+H].sup.+=215.09.
Step C:
(R)--N-(6-(Pyridin-4-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine
##STR00810##
[0533] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.059 g, 0.257 mmol) in N,N-dimethylformamide (15 mL) was added
Cs.sub.2CO.sub.3 (0.209 g, 0.642 mmol) and
4-isothiocyanato-6-(pyridin-4-yl)pyrimidine (0.055 g, 0.257 mmol).
The suspension was stirred at room temperature for 30 minutes.
N,N'-diisopropylcarbodiimide (0.12 mL, 0.77 mmol) was then added
and the mixture was continued to stir at room temperature for 18
hours. The mixture was concentrated and purified by silica gel
chromatography (0-10% [9:1 methanol:ammonium hydroxide]-ethyl
acetate) to afford
(R)--N-(6-(pyridin-4-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine (0.014 g, 0.04 mmol, 16% yield) as a
yellow film. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.85 (1H, d),
8.67 (2H, dd), 8.04 (2H, dd), 7.34 (1H, br. s.), 4.06 (1H, d), 3.76
(1H, d), 3.23 (1H, d), 3.10 (1H, d), 2.70-2.99 (4H, m), 2.01-2.22
(2H, m), 1.53-1.86 (3H, m). LCMS R.T.=0.42; [M+H].sup.+=337.14.
Example 247
(R)-6-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)-2-meth-
ylnicotinonitrile
##STR00811##
[0534] Step A: 6-Isothiocyanato-2-methylnicotinonitrile
##STR00812##
[0536] To 6-amino-2-methylnicotinonitrile (0.41 g, 3.08 mmol) in
dichloromethane (20 mL) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (0.75 g, 3.23 mmol). The
reaction was stirred at 40.degree. C. for 3 hours. The reaction was
cooled to room temperature. The crude was purified by
chromatography (Biotage: 25-100% ethyl acetate/hexane) to yield
6-isothiocyanato-2-methylnicotinonitrile (0.52 g, 2.97 mmol, 96%
yield). 1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.36 (d, J=8.24 Hz,
1H), 7.39 (d, J=8.24 Hz, 1H), 2.65 (s, 3H). MS (LC/MS) R.T.=2.09;
[M+H].sup.+=176.0.
Step B:
(R)-6-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino-
)-2-methylnicotinonitrile
##STR00813##
[0538] To 6-isothiocyanato-2-methylnicotinonitrile (0.25 g, 1.43
mmol) in N,N-dimethylformamide (20 mL) was added triethylamine (0.5
mL, 3.666 mmol) and 3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.33 g, 1.46 mmol) at room temperature. The reaction was stirred
at 70.degree. C. for 2 hours. The reaction was cooled to room
temperature and concentrated in vacuo. The crude urea was purified
by chromatography (Biotage: 85% CHCl.sub.3, 14% MeOH, 1%
NH.sub.4OH). The product was then treated with
N,N-dimethylformamide (20 mL) and N,N'-diisopropylcarbodiimide
(0.67 mL, 4.28 mmol). The reaction was heated to 70.degree. C. for
2 hours. The reaction was cooled to room temperature and
concentrated in vacuo to yield the crude product. The crude product
was purified by chromatography (Biotage: 85% CHCl.sub.3, 14% MeOH,
1% NH.sub.4OH) to yield
(R)-6-(4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)-2-met-
hylnicotinonitrile (0.09 g, 0.3 mmol, 21% yield) as a white powder.
1H NMR (500 MHz, DMSO-D6) .delta. ppm 9.11 (s, 1H), 7.87 (d, J=7.93
Hz, 2H), 6.69 (s, 1H), 3.89 (d, J=10.38 Hz, 2H), 3.63 (d, J=10.38
Hz, 3H), 3.00 (s, 5H), 2.72-2.80 (m, 4H), 2.64-2.69 (m, 5H), 2.60
(s, 7H), 2.00 (d, J=2.14 Hz, 3H), 1.91 (s, 1H), 1.87 (s, 2H), 1.58
(s, 5H), 1.42-1.50 (m, 2H). MS (LC/MS) R.T.=0.48;
[M+H].sup.+=298.13.
Example 248
(R)-6-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)-2,4-di-
methylnicotinonitrile
##STR00814##
[0539] Step A: 6-Isothiocyanato-2,4-dimethylnicotinonitrile
##STR00815##
[0541] To 6-amino-2,4-dimethylnicotinonitrile (0.14 g, 0.95 mmol)
in dichloromethane (20 mL) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (0.23 g, 0.1 mmol). The
reaction was stirred at 40.degree. C. for 3 hours. The reaction was
cooled to room temperature. The crude was purified by
chromatography (Biotage: 25-100% ethyl acetate/hexane) to yield
6-isothiocyanato-2,4-dimethylnicotinonitrile (0.15 g, 0.79 mmol,
83% yield). 1H NMR (500 MHz, DMSO-D6) .delta. ppm 7.37 (s, 1H),
2.63 (s, 3H). MS (LC/MS) R.T.=2.40; [M+H].sup.+=190.
Step B:
(R)-6-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino-
)-2,4-dimethylnicotinonitrile
##STR00816##
[0543] To 6-isothiocyanato-2,4-dimethylnicotinonitrile (0.09 g,
0.48 mmol) in N,N-dimethylformamide (20 mL) was added triethylamine
(0.17 mL, 1.19 mmol) and 3-(aminomethyl)quinuclidin-3-ol
dihydrochloride (0.11 g, 0.49 mmol) at room temperature. The
reaction was stirred at 70.degree. C. for 2 hours. The reaction was
cooled to room temperature and concentrated in vacuo. The crude
urea was purified by chromatography (Biotage: 85% CHCl.sub.3, 14%
MeOH, 1% NH.sub.4OH). The product was then treated with
N,N-dimethylformamide (20 mL) and N,N'-diisopropylcarbodiimide
(0.22 mL, 1.43 mmol). The reaction was heated to 70.degree. C. for
2 hours. The reaction was cooled to room temperature and
concentrated in vacuo to yield the crude product. The crude product
was purified by chromatography (Biotage: 85% CHCl.sub.3, 14% MeOH,
1% NH.sub.4OH) to yield
(R)-6-(4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octane]-2-ylamino)-2,4-d-
imethylnicotinonitrile (0.10 g, 0.32 mmol, 66% yield) as a white
powder. 1H NMR (500 MHz, DMSO-D6) .delta. ppm 9.08 (s, 1H), 6.60
(s, 1H), 3.88 (d, J=10.38 Hz, 1H), 3.61 (d, J=10.38 Hz, 1H), 2.98
(s, 2H), 2.70-2.79 (m, 2H), 2.63-2.69 (m, 2H), 2.55-2.60 (m, 4 H),
2.31-2.39 (m, 4H), 1.99 (s, 1H), 1.89 (s, 1H), 1.54-1.62 (m, 2H),
1.41-1.49 (m, 1H). MS (LC/MS) R.T.=0.78; [M+H].sup.+=312.1.
Example 249
(R)--N-(6-Phenylpyridazin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]-
octan]-2-amine
##STR00817##
[0544] Step A: 3-Isothiocyanato-6-phenylpyridazine
##STR00818##
[0546] 3-Isothiocyanato-6-phenylpyridazine was synthesized by the
method of Example 23, Step B. Flash chromatography on a 120 g
silica gel cartridge with 0 to 25% EtOAc in hexane, 25 min, at 35
mL/min afforded 420 mg (49% yield).
[0547] LCMS: RT=2.17 min, MH+=214.06.
Step B:
(R)--N-(6-Phenylpyridazin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine
##STR00819##
[0549]
(R)--N-(6-Phenylpyridazin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine was synthesized by the method of Example 23,
Step B. Flash chromatography on a 160 g silica gel cartridge with
1-4% [9:1 MeOH/NH.sub.4OH] in CHCl.sub.3, 50 min, at 40 mL/min
afforded 67 mg
(R)--N-(6-phenylpyridazin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine (17% yield).
[0550] 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.61 (1H, br. s.),
7.95-7.99 (2H, m), 7.74 (1 H, d, J=9.32 Hz), 7.39-7.52 (3H, m),
7.22 (1H, partial d), 3.97 (1H, d, J=9.32 Hz), 3.64 (1H, d, J=9.32
Hz), 3.37 (1H, dd, J=14.73, 1.38 Hz), 2.69-3.06 (5H, m), 2.16-2.26
(1H, m), 2.14 (1H, br. s.), 1.66-1.79 (1H, m), 1.45-1.60 (2H,
m)
[0551] 1H NMR (400 MHz, MeOD) .delta. ppm 7.87-8.02 (3H, m),
7.44-7.55 (3H, m), 7.13-7.29 (1H, m), 4.05 (1H, d, J=9.82 Hz), 3.74
(1H, d, J=10.07 Hz), 3.17 (2H, dd, J=49.35, 14.60 Hz), 2.73-3.04
(4H, m), 2.16 (2H, br. s.), 1.55-1.85 (3H, m) LCMS: RT=0.82 min,
MH-=334.2, MH+=336.2.
Example 250
(R)--N-(5-(Methylthio)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.-
2.2]octan]-2-amine
##STR00820##
[0552] Step A: 5-(Methylthio)pyrazin-2-amine
##STR00821##
[0554] To a solution of 5-bromopyrazin-2-amine (2 g, 11.49 mmol) in
N,N-dimethylformamide (20 ml) was added sodium thiomethoxide (1.611
g, 22.99 mmol). The mixture was stirred and heated at 100.degree.
C. under nitrogen for 18 h and concentrated. The residue was
treated with water and the mixture was extracted with
dichloromethane. The combined organics were dried with sodium
sulfate, filtered and concentrated. The residue was purified by
silica gel chromatography (0-10% 9:1-methanol:ammonium
hydroxide-ethyl acetate) to afford
6-(pyridin-3-yl)pyrimidin-4-amine (0.15 g, 0.871 mmol, 35% yield)
as a yellow solid. LCMS R.T.=0.91; [M+H].sup.+=141.89.
Step B: 2-Isothiocyanato-5-(methylthio)pyrazine
##STR00822##
[0556] To a solution of 1,1'-thiocarbonyldipyridin-2(1H)-one (1.069
g, 4.60 mmol) in dichloromethane at room temperature was added
5-(methylthio)pyrazin-2-amine (0.50 g, 3.54 mmol). The reaction was
stirred at room temperature for 18 hours. LC/MS showed the desired
product peak as the major peak. The deep orange mixture was
purified by silica gel chromatography (0-40% ethyl acetate-hexanes)
to afford 2-isothiocyanato-5-(methylthio)pyrazine (0.545 g, 0.257
mmol, 84% yield) as an orange oil. LCMS R.T.=2.65;
[M+H].sup.+=184.02.
Step C:
(R)--N-(5-(Methylthio)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine
##STR00823##
[0558] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.375 g, 1.637 mmol) in N,N-dimethylformamide (15 mL) was added
Cs.sub.2CO.sub.3 (1.333 g, 4.09 mmol) and
2-isothiocyanato-5-(methylthio)pyrazine (0.3 g, 1.637 mmol). The
suspension was stirred at room temperature for 30 minutes.
N,N'-diisopropylcarbodiimide (0.765 mL, 4.9 mmol) was then added
and the mixture was continued to stir at room temperature for 18
hours. The mixture was concentrated and purified by silica gel
chromatography (5-25% 9:1 methanol:ammonium hydroxide-ethyl
acetate) to afford
(R)--N-(5-(methylthio)pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine (0.014 g, 0.04 mmol, 16% yield) as a yellow
solid. M.P. 155-60.degree. C. .sup.1H NMR (400 MHz, MeOD) .delta.
ppm 8.13 (1H, d), 8.06 (1H, s), 3.96 (1H, d), 3.66 (1H, d), 3.20
(1H, d), 3.08 (1H, d), 2.87-2.96 (2H, m), 2.72-2.82 (2H, m), 2.52
(3H, s), 2.00-2.19 (2H, m), 1.51-1.81 (3H, m). LCMS R.T.=1.01;
[M+H].sup.+=306.12.
Example 251
(R)--N-(5,6-Dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00824##
[0559] Step A: N-(5,6-Dichloropyridin-2-yl)pivalamide
##STR00825##
[0561] To a solution of N-(6-chloropyridin-2-yl)pivalamide,
synthesized as in J. Org. Chem. 2005, 70, 1771, (1.02 g, 4.80 mmol)
in chloroform (25 mL) was added 1-chloropyrrolidine-2,5-dione (0.62
g, 4.67 mmol) and the mixture was refluxed in an oil bath for 3
hrs. It was allowed to cool to room temperature overnight. The
reaction mixture was evaporated in vacuo and re-dissolved in DMF
(15 mL). Another 480 mg 1-chloropyrrolidine-2,5-dione was added and
the resulting solution was heated overnight in an oil bath at
95-100.degree., then cooled again to room temperature. The solvent
was removed in vacuo and the residue was partitioned between water
and ethyl acetate. The aqueous phase was washed twice more with
ethyl acetate and the combined organic phases were washed with
brine, dried over magnesium sulfate, and evaporated in vacuo. TLC
(10% ethyl acetate/hexane) showed a robust spot at Rf 0.6 with
smaller spots at Rf 0.4 and 0.2. The material was subjected to the
Biotage in 5-10% ethyl acetate/hexane, collecting the Rf 0.6
fraction to give 790 mg (66%) white solid,
N-(5,6-dichloropyridin-2-yl)pivalamide. 1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 8.17 (s, 1H), 7.96 (s, 1H), 7.72 (s, 1H),
1.31 (s, 10H). MS (LC/MS) R.T.=1.85; [M+H].sup.+=248.8.
Step B: 5,6-Dichloropyridin-2-amine
##STR00826##
[0563] A mixture of N-(5,6-dichloropyridin-2-yl)pivalamide (790 mg,
3.20 mmol), hydrochloric acid, 37% (1.25 mL), water (1.25 mL), and
EtOH (3 mL) was heated for 4 hrs in an oil bath at 85-90.degree. C.
LCMS showed nearly complete conversion to product. The reaction was
cooled to room temperature and the reaction mixture was evaporated
down to a small volume, then transferred to a separatory funnel
where it was partitioned between aqueous sodium carbonate and ethyl
acetate. The layers were separated, the aqueous phase was washed
again with ethyl acetate, and the combined organic phases were
washed with brine, dried over MgSO.sub.4, filtered, and evaporated
to give a white solid. The material was subjected to a Biotage
column in 20% ethyl acetate/hexane, collecting the main component.
5,6-Dichloropyridin-2-amine (0.49 g, 2.98 mmol, 93%) was obtained
as a white solid. 1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 7.44 (d,
J=8.55 Hz, 1H), 6.36 (d, J=8.24 Hz, 1H), 4.58 (s, 2H). MS (LC/MS)
R.T.=1.28; [M+H].sup.+=164.8.
Step C: 5,6-Dichloro-2-isothiocyanatopyridine
##STR00827##
[0565] To 5,6-dichloropyridin-2-amine (0.47 g, 2.88 mmol) in
dichloromethane (25 mL) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (0.68 g, 2.94 mmol). The
reaction was stirred at 40.degree. C. for 3 hours, then cooled to
room temperature. The crude material was purified by chromatography
(Biotage: 25-100% ethyl acetate/hexane)
2,3-dichloro-6-isothiocyanatopyridine (0.48 g, 2.34 mmol, 81%
yield) as a white powder. 1H NMR (500 MHz, DMSO-D6) .delta. ppm
8.26 (d, J=8.55 Hz, 1H), 7.47 (d, J=8.24 Hz, 1H). MS (LC/MS)
R.T.=2.83; [M+H].sup.+=204.8.
Step D:
(R)--N-(5,6-Dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00828##
[0567] To 2,3-dichloro-6-isothiocyanatopyridine (0.47 g, 2.29 mmol)
in N,N-dimethylformamide (20 mL) was added triethylamine (0.8 mL,
5.7 mmol) and 3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.54
g, 2.34 mmol) at room temperature. The reaction was stirred at
70.degree. C. for 2 hours, cooled to room temperature and
concentrated in vacuo. The crude urea was purified by
chromatography (Biotage: 85% CHCl.sub.3, 14% MeOH, 1% NH.sub.4OH).
The product was then treated with N,N-dimethylformamide (20 mL) and
N,N'-diisopropylcarbodiimide (1.07 mL, 6.88 mmol). The reaction was
heated to 70.degree. C. for 2 hours. The reaction was cooled to
room temperature and concentrated in vacuo. The crude product was
purified by chromatography (Biotage: 85% CHCl.sub.3, 14% MeOH, 1%
NH.sub.4OH) to yield
(R)--N-(5,6-dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2-
.2]octan]-2-amine (0.36 g, 1.08 mmol, 47% yield) as a white powder.
1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.31 (d, J=1.22 Hz, 1H), 7.84
(s, 1H), 6.80 (s, 1H), 3.85 (d, J=10.07 Hz, 1H), 3.57 (d, J=10.38
Hz, 2H), 2.98 (s, 3H), 2.69-2.78 (m, 3H), 2.65 (t, J=7.78 Hz, 3H),
2.00 (s, 2H), 1.86 (s, 2 H), 1.58 (dd, J=7.48, 2.90 Hz, 2H), 1.56
(s, 1H), 1.41-1.49 (m, 2H). MS (LC/MS) R.T.=0.81;
[M+H].sup.+=327.1.
Example 252
(R)--N-(4,5-Dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00829##
[0568] Step A: 4,5-Dichloro-2-isothiocyanatopyridine
##STR00830##
[0570] To 4,5-dichloropyridin-2-amine (0.25 g, 1.53 mmol) in
dichloromethane (25 mL) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (0.36 g, 1.56 mmol). The
reaction was stirred at 40.degree. C. for 3 hours, then cooled to
room temperature. The crude material was purified by chromatography
(Biotage: 25-100% ethyl acetate/hexane) to yield
4,5-dichloro-2-isothiocyanatopyridine (0.26 g, 1.27 mmol, 83%
yield) as a yellow powder. The product was carried directly to the
next step.
Step B:
(R)--N-(4,5-Dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00831##
[0572] To 4,5-dichloro-2-isothiocyanatopyridine (0.25 g, 1.22 mmol)
in N,N-dimethylformamide (20 mL) was added triethylamine (0.43 mL,
3.05 mmol) and (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.29 g, 1.24 mmol) at room temperature. The reaction was stirred
at 70.degree. C. for 2 hours. The reaction was cooled to room
temperature and concentrated in vacuo. The crude urea was purified
by chromatography (Biotage: 85% CHCl.sub.3, 14% MeOH, 1%
NH.sub.4OH). The product was then treated with
N,N-dimethylformamide (20 mL) and N,N'-diisopropylcarbodiimide
(0.57 mL, 3.66 mmol). The reaction was heated to 70.degree. C. for
2 hours. The reaction was cooled to room temperature and
concentrated to yield the crude product. The crude product was
purified by chromatography (Biotage: 85% CHCl.sub.3, 14% MeOH, 1%
NH.sub.4OH) to yield
(R)--N-(4,5-dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2-
.2]octan]-2-amine (0.09 g, 0.27 mmol, 22% yield) as a white powder.
1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.81 (s, 1H), 8.32 (d, J=6.10
Hz, 1H), 7.03 (s, 1H), 3.83 (d, J=9.46 Hz, 1H), 3.57 (d, J=9.77 Hz,
1H), 2.98 (s, 2H), 2.71-2.79 (m, 2H), 2.65 (t, J=7.78 Hz, 2H), 1.99
(s, 1H), 1.86 (s, 1 H), 1.53-1.61 (m, 2H), 1.41-1.49 (m, 1H). MS
(LC/MS) R.T.=0.78; [M+H].sup.+=327.0.
Example 253
(R)--N-(5-Chloro-4-methylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine
##STR00832##
[0573] Step A: 5-Chloro-2-isothiocyanato-4-methylpyridine
##STR00833##
[0575] To 5-chloro-4-methylpyridin-2-amine (0.41 g, 2.88 mmol) in
dichloromethane (25 mL) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (0.70 g, 3.0 mmol). The
reaction was stirred at 40.degree. C. for 3 hours. The reaction was
cooled to room temperature. The crude mixture was purified by
chromatography (Biotage: 25-100% ethyl acetate/hexane) to yield
5-chloro-2-isothiocyanato-4-methylpyridine (0.45 g, 2.44 mmol, 85%
yield). 1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.45 (s, 1H), 7.47
(s, 1H), 2.37 (s, 3H). LC/MS RT=2.79; [M+H].sup.+=184.9.
Step B:
(R)--N-(5-Chloro-4-methylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octan]-2-amine
##STR00834##
[0577] To 5-chloro-2-isothiocyanato-4-methylpyridine (0.37 g, 2.0
mmol) in N,N-dimethylformamide (20 mL) was added triethylamine (0.7
mL, 5.0 mmol) and (S)-3-(aminomethyl)quinuclidin-3-ol
dihydrochloride (from Step B of Example 17) (0.47 g, 2.0 mmol) at
room temperature. The reaction was stirred at 70.degree. C. for 2
hours. The reaction was cooled to room temperature and concentrated
in vacuo. The crude urea was purified by chromatography (Biotage:
85% CHCl.sub.3, 14% MeOH, 1% NH.sub.4OH). The product was then
treated with N,N-dimethylformamide (20 mL) and
N,N'-diisopropylcarbodiimide (0.94 mL, 6.0 mmol). The reaction was
heated to 70.degree. C. for 2 hours. The reaction was cooled to
room temperature and concentrated to yield the crude product. The
crude product was purified by chromatography (Biotage: 85%
CHCl.sub.3, 14% MeOH, 1% NH.sub.4OH) to yield
(R)--N-(5-chloro-4-methylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine (0.19 g, 0.61 mmol, 30.3% yield) as a white
powder. 1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.79 (s, 1H),
8.08-8.15 (m, 2H), 6.78 (s, 1H), 3.82 (d, J=8.55 Hz, 2H), 3.55 (d,
J=10.38 Hz, 2 H), 2.93-3.02 (m, 5H), 2.71-2.80 (m, 5H), 2.66 (t,
J=7.63 Hz, 4H), 2.23-2.29 (m, 7H), 1.94-2.02 (m, 2H), 1.92 (s, 1H),
1.86 (s, 2H), 1.53-1.62 (m, 5H), 1.41-1.49 (m, J=12.55, 9.88, 7.02,
2.29 Hz, 2H). MS (LC/MS) R.T.=0.72; [M+H].sup.+=307.1.
Example 254
(R)--N-(6-Chloropyridazin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]-
octan]-2-amine
##STR00835##
[0578] Step A: 3-Chloro-6-isothiocyanatopyridazine
##STR00836##
[0580] 3-Chloro-6-isothiocyanatopyridazine was synthesized by the
method of Example 218, Step D. Flash chromatography on a 120 g
silica gel cartridge with 0 to 25% EtOAc in hexane, 25 min, at 35
mL/min afforded 213 mg (31% yield).
[0581] LCMS: RT=1.25 min, MH+=172.00.
Step B:
(R)--N-(6-Chloropyridazin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine
##STR00837##
[0583]
(R)--N-(6-Chloropyridazin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine was synthesized by the method of Example 218,
Step E. Flash chromatography on a 160 g silica gel cartridge with
1-3% [9:1 MeOH/NH4OH] in CHCl3, 50 min, at 40 mL/min afforded 29 mg
(8% yield).
[0584] 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.27 (1H, br. s.),
7.28 (1H, d, J=9.07 Hz), 7.10 (1H, d, J=9.07 Hz), 3.95 (1H, d,
J=9.57 Hz), 3.62 (1H, d, J=9.57 Hz), 3.34 (1H, dd, J=14.98, 1.64
Hz), 2.67-3.04 (5H, m), 2.14-2.21 (1H, m), 2.12 (1H, br. s.),
1.65-1.79 (1H, m), 1.45-1.61 (2H, m).
[0585] LCMS: RT=0.62 min, MH-=292.1, MH+=294.1.
Example 255
(R)--N-(6-Bromopyridazin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]o-
ctan]-2-amine
##STR00838##
[0586] Step A: 3-Bromo-6-isothiocyanatopyridazine
##STR00839##
[0588] 3-Bromo-6-isothiocyanatopyridazine was synthesized by the
method of Example 218, Step D. Flash chromatography on a 120 g
silica gel cartridge with 0 to 25% EtOAc in hexane, 25 min, at 35
mL/min afforded 364 mg (42% yield).
[0589] LCMS: RT=1.34 min, MH+=215.92.
Step B:
(R)--N-(6-Bromopyridazin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine
##STR00840##
[0591]
(R)--N-(6-Bromopyridazin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine was synthesized by the method of Example 218,
Step E. Flash chromatography on a 160 g silica gel cartridge with
1-3% [9:1 MeOH/NH4OH] in CHCl3, 50 min, at 40 mL/min afforded 211
mg (37% yield). 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.23 (1H,
br. s.), 7.37 (1H, d, J=9.07 Hz), 6.97 (1H, d, J=9.07 Hz), 3.92
(1H, d, J=9.57 Hz), 3.59 (1H, d, J=9.82 Hz), 3.29 (1H, dd, J=14.98,
1.64 Hz), 2.63-2.99 (5H, m), 2.04-2.19 (2H, m), 1.59-1.74 (1H, m),
1.39-1.58 (2H, m)
[0592] LCMS: RT=0.64 min., MH-336.1, MH+338.0.
Example 256
(R)--N-(6-(4-Chlorophenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00841##
[0593] Step A: 6-(Pyridin-4-yl)pyrimidin-4-amine
##STR00842##
[0595] A mixture of 6-chloropyrimidin-4-amine (0.324 g, 2.5 mmol),
4-chlorophenylboronic acid (0.489 g, 3.13 mmol), Na.sub.2CO.sub.3
(0.795 g, 7.50 mmol) and bis(triphenylphosphine)palladium(II)
chloride (0.035 g, 0.050 mmol) was suspended in a mixture of
DME/EtOH/water (15:2:3 mL). The mixture was heated in the microwave
synthesizer at 125.degree. C. for 20 min and concentrated. The
residue was purified by silica gel chromatography (2-15% 9:1
methanol:ammonium hydroxide-ethyl acetate) to afford:
6-(pyridin-3-yl)pyrimidin-4-amine (0.3 g, 0.871 mmol, 58.4% yield)
as an off-white solid. LCMS R.T.=1.42; [M+2H].sup.+=207.91.
Step B: 4-(4-Chlorophenyl)-6-isothiocyanatopyrimidine
##STR00843##
[0597] To a bright orange solution of
1,1'-thiocarbonyldipyridin-2(1H)-one (0.666 g, 2.87 mmol)
dichloromethane/N,N-dimethylformamide at room temperature was added
6-(4-chlorophenyl)pyrimidin-4-amine (0.59 g, 2.87 mmol). The orange
mixture was heated at 60.degree. C. for 18 hours. The LC/MS showed
the desired product peak as a major peak. The deep orange mixture
was purified by silica gel chromatography (0-40% ethyl
acetate-hexanes) to afford
4-(4-chlorophenyl)-6-isothiocyanatopyrimidine (0.322 g, 1.300 mmol,
45% yield) as an orange oil. LCMS R.T.=2.82; [M].sup.+=248.03.
Step C:
(R)--N-(6-(4-Chlorophenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR00844##
[0599] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.298 g, 1.300 mmol) in N,N-dimethylformamide (15 mL) was added
Cs.sub.2CO.sub.3 (1.059 g, 3.25 mmol) and
4-(4-chlorophenyl)-6-isothiocyanatopyrimidine (0.322 g, 1.300
mmol). The suspension was stirred at room temperature for 30
minutes. N,N'-Diisopropylcarbodiimide (0.608 mL, 3.90 mmol) was
then added and the mixture was stirred at room temperature for 18
hours. The mixture was concentrated and purified by silica gel
chromatography (5-25% 9:1 methanol:ammonium hydroxide-ethyl
acetate) to afford
(R)--N-(6-(pyridin-4-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine (0.104 g, 0.276 mmol, 21% yield) as a
yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.54
(1H, br. s.), 8.83 (1H, d), 7.89-8.04 (2H, m), 7.41-7.54 (2H, m),
7.33 (1H, br. s.), 4.02 (1H, d), 3.71 (1H, d), 3.42 (1H, d),
2.73-3.15 (5H, m), 2.10-2.31 (2H, m), 1.46-1.89 (3 H, m). LCMS
R.T.=1.92; [M].sup.+=370.35.
Example 257
(R)--N-(6-(3-Chlorophenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00845##
[0600] Step A: 6-(3-Chlorophenyl)pyrimidin-4-amine
##STR00846##
[0602] A mixture of 6-chloropyrimidin-4-amine (0.324 g, 2.5 mmol),
3-chlorophenylboronic acid (0.489 g, 3.13 mmol), Na.sub.2CO.sub.3
(0.795 g, 7.50 mmol) and bis(triphenylphosphine)palladium(II)
chloride (0.035 g, 0.050 mmol) was suspended in a mixture of
DME/EtOH/wate (15:2:3 mL). The mixture was heated in the microwave
synthesizer at 125.degree. C. for 20 min and concentrated. The
residue was purified by silica gel chromatography (30-70% ethyl
acetate in hexanes) to afford: 6-(3-chlorophenyl)pyrimidin-4-amine
(0.47 g, 2.286 mmol, 91% yield) as a yellow solid. LCMS R.T.=1.45;
[M+2H].sup.+=208.05.
Step B: 4-(3-Chlorophenyl)-6-isothiocyanatopyrimidine
##STR00847##
[0604] To a bright orange solution of
1,1'-thiocarbonyldipyridin-2(1H)-one (0.486 g, 2.091 mmol) in
dichloromethane/N,N-dimethylformamide at room temperature was added
6-(3-chlorophenyl)pyrimidin-4-amine (0.43 g, 2.091 mmol). The
orange mixture was heated at 60.degree. C. for 18 hours. The LC/MS
showed the desired product peak as a major peak. The deep orange
mixture was purified by silica gel chromatography (0-40% ethyl
acetate-hexanes) to afford
4-(3-chlorophenyl)-6-isothiocyanatopyrimidine (0.12 g, 0.484 mmol,
23% yield) as an orange oil. LCMS R.T.=2.15; [M].sup.+=248.31.
Step C:
(R)--N-(6-(3-Chlorophenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR00848##
[0606] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.111 g, 0.484 mmol) in N,N-dimethylformamide (15 mL) was added
Cs.sub.2CO.sub.3 (0.395 g, 1.211 mmol) and
4-(3-chlorophenyl)-6-isothiocyanatopyrimidine (0.12 g, 0.484 mmol).
The suspension was stirred at room temperature for 30 minutes.
N,N'-diisopropylcarbodiimide (0.226 mL, 1.453 mmol) was then added
and the mixture was continued to stir at room temperature for 18
hours. The mixture was concentrated and purified by silica gel
chromatography (5-25%, then, 2-10% 9:1 methanol:ammonium
hydroxide-ethyl acetate) to afford
(R)--N-(6-(3-chlorophenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine (0.086 g, 0.221 mmol, 46% yield) as a
yellow solid. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.82 (1H, d),
8.05 (1H, d), 7.93 (1H, ddd), 7.43-7.52 (2H, m), 7.35 (1H, br. s.),
4.13 (1H, d), 3.93 (1H, d), 3.63-3.81 (2H, m), 3.42-3.53 (1H, m),
3.30-3.40 (3H, m), 2.46 (1H, d), 2.26-2.40 (1H, m), 1.88-2.16 (3H,
m). LCMS R.T.=1.90; [M].sup.+=370.28.
Example 258
(R)--N-(5-Methyl-1,3,4-oxadiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine
##STR00849##
[0607] Step A: Dimethyl
5-methyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate
##STR00850##
[0609] To a solution of 5-methyl-1,3,4-oxadiazol-2-amine (1.92 g,
20 mmol) in DMF (10 ml) was added NaOH (20M, 2 ml), CS.sub.2 (3
ml), NaOH (20M, 2 ml) and iodomethane (3 ml) slowly over 10
minutes. The mixture was stirred at room temperature for 1 h and
poured into 20 ml water. The precipitated solid was filtered,
washed with water, and dried to obtain the desired product,
dimethyl 5-methyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate as a
white solid (1.45 g, 35.7%). .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. ppm 2.63 (s, 6H), 2.50 (s, 3H). LCMS R.T. 1.66 min;
[M+H]=203.91.
Step B:
(R)--N-(5-Methyl-1,3,4-oxadiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octan]-2-amine
##STR00851##
[0611] A mixture of dimethyl
5-methyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate (260 mg, 1.28
mmol), (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (200 mg,
1.28 mmol) and cesium carbonate (876 mg, 2.69 mmol) in DMF (5 ml)
was stirred overnight at room temperature. The mixture was
concentrated and purified on a Biotage silica gel column (100%
ethyl acetate, then 10-35% 9:1 methanol:ammonium
hydroxide-chloroform) to obtain the desired product,
(R)--N-(5-methyl-1,3,4-oxadiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine (192 mg, 54.1%). .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 4.05 (d, 1H), 3.74 (d, 1H), 3.25 (d, 1H), 3.15 (d, 1H),
2.94 (m, 2H), 2.85 (m, 2H), 2.43 (s, 3H), 2.19 (m, 1H), 2.10 (m,
1H), 1.6-1.8 (m, 3H). MS (LCMS) [M+H]=264.05. R.T. 0.16 min.
Example 259
(R)--N-(3-Methyl-1,2,4-thiadiazol-5-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine
##STR00852##
[0612] Step A: Dimethyl
3-methyl-1,2,4-thiadiazol-5-ylcarbonimidodithioate
##STR00853##
[0614] To a solution of 3-methyl-1,2,4-thiadiazol-5-amine (2.3 g,
20 mmol) in DMF (10 ml) was added NaOH (20M, 2 ml), CS.sub.2 (3
ml), NaOH (20M, 2 ml) and iodomethane (3 ml) slowly over 10
minutes. The mixture was stirred at room temperature for 1 h and
poured into 20 ml water. The precipitated solid was filtered and
washed with water, and dried to obtain impure dimethyl
3-methyl-1,2,4-thiadiazol-5-ylcarbonimidodithioate, a yellow solid
(2.3 g, 52.5%). .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 2.67
(s), 2.62 (s). MS [M+H]=219.85.
Step B:
(R)--N-(3-Methyl-1,2,4-thiadiazol-5-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine
##STR00854##
[0616] A mixture of dimethyl
3-methyl-1,2,4-thiadiazol-5-ylcarbonimidodithioate (281 mg, 1.28
mmol), (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (200 mg,
1.28 mmol) and cesium carbonate (876 mg, 2.69 mmol) in DMF (5 ml)
was stirred overnight at room temperature. The mixture was
concentrated and purified on a Biotage silica gel column (100%
ethyl acetate, then 10-35% 9:1 methanol:ammonium
hydroxide-chloroform) to obtain the desired product,
(R)--N-(3-methyl-1,2,4-thiadiazol-5-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine (147.8 mg, 40.5%). .sup.1H NMR (500 MHz,
MeOD) .delta. ppm 4.01-3.99 (d, 1H), 3.72-3.70 (d, 1H), 3.27 (d,
1H), 3.16 (d, 1H), 3.01-2.9 (m, 2H), 2.86-2.83 (m, 2H), 2.43 (s,
3H), 2.21-2.0 (m, 2H), 1.81-1.75 (m, 1H), 1.75-1.70 (m, 2H). MS
(LCMS) [M+H]=279.99; R.T.=0.2 min.
Example 260
(R)--N-(3-Methyl-1,2,4-oxadiazol-5-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine
##STR00855##
[0617] Step A: Dimethyl
3-methyl-1,2,4-oxadiazol-5-ylcarbonimidodithioate
##STR00856##
[0619] To a solution of 3-methyl-1,2,4-oxadiazol-5-amine (490 mg,
4.94 mmol) in DMF (5 ml) was added NaOH (20M, 0.5 ml), CS.sub.2 (1
ml), NaOH (20M, 0.5 ml) and iodomethane (1 ml) slowly over 10
minutes. The mixture was stirred at room temperature for 1 hour.
The mixture became very thick, and 20 ml water was added. The solid
was filtered off, washed with water, and dried to obtain impure
dimethyl 3-methyl-1,2,4-oxadiazol-5-ylcarbonimidodithioate, a
yellow solid (770 mg, 77%). MS (LCMS) [M+H]=203.91; R.T.=1.84 min.
The product was used directly in the next step.
Step B:
(R)--N-(3-Methyl-1,2,4-oxadiazol-5-yl)-4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octan]-2-amine
##STR00857##
[0621] A mixture of dimethyl
3-methyl-1,2,4-oxadiazol-5-ylcarbonimidodithioate (280 mg, 1.37
mmol), (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (215 mg,
1.37 mmol) and cesium carbonate (942 mg, 2.89 mmol) in DMF (5 ml)
was stirred overnight at room temperature. The mixture was
concentrated and purified on a Biotage silica gel column (100%
ethyl acetate, then 10-35% 9:1 methanol:ammonium
hydroxide-chloroform) to obtain the desired product,
(R)--N-(3-methyl-1,2,4-oxadiazol-5-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine (198 mg, 51.9%). .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 4.1-4.0 (d, 1H), 3.8-3.7 (d, 1H), 3.4-3.2 (d, 1H),
3.2-3.1 (d, 1H), 3.0-2.9 (m, 2H), 2.9-2.8 (m, 2H), 2.27 (s, 3H),
2.2 (m, 1H), 2.2-2.0 (m, 1H), 1.9-1.6 (m, 3H). MS (LCMS)
[M+H]=264.05; R.T.=0.26 min.
Example 261
(R)--N-(6-(Methylthio)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine
##STR00858##
[0622] Step A: Dimethyl
6-chloropyrimidin-4-ylcarbonimidodithioate
##STR00859##
[0624] To a solution of 6-chloropyrimidin-4-amine (1.295 g, 10
mmol) in N,N-dimethylformamide (12 mL) was added dropwise NaOH (1
mL, 20.00 mmol, 20 M), CS2 (1.5 mL, 24.88 mmol), NaOH (1 mL, 20.00
mmol, 20 M) and iodomethane (1.5 mL, 23.99 mmol) at 15 min
intervals. Stirring was continued for 1.5 h and the mixture was
poured into water. The orange solid was separated washed with
water, dried and recrystallised from methanol to afford dimethyl
6-chloropyrimidin-4-ylcarbonimidodithioate (0.966 g, 4.13 mmol,
41.3% yield) as a yellow solid. LCMS R.T.=2.39;
[M+H].sup.+=234.08.
Step B:
(R)--N-(6-(Methylthio)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine
##STR00860##
[0626] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.9
g, 3.93 mmol) in N,N-dimethylformamide (20 mL) was added
Cs.sub.2CO.sub.3 (2.69 g, 8.25 mmol) and dimethyl
6-chloropyrimidin-4-ylcarbonimidodithioate (0.964 g, 4.12 mmol).
The suspension was stirred at room temperature for 18 hours, then
heated at 100 C for 3 hours. The mixture was concentrated and
purified by silica gel chromatography (5-15% 9:1 methanol:ammonium
hydroxide-ethyl acetate) to afford
(R)--N-(6-(methylthio)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine (0.21 g, 0.72 mmol, 48.2% yield) as a pale
yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.43
(1H, br. s.), 8.55 (1H, d), 6.78 (1H, br. s.), 3.98 (1H, d), 3.64
(1H, d), 3.37 (1H, dd), 2.72-3.06 (5H, m), 2.51 (3H, s), 2.08-2.24
(2H, m), 1.69-1.81 (1H, m), 1.41-1.64 (2H, m). LCMS R.T.=0.93;
[M+H].sup.+=306.29.
Example 262
(R)--N-([1,2,4]Triazol[4,3-a]pyridine-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine
##STR00861##
[0627] Step A: Di(1H-imidazol-1-yl)methanimine
##STR00862##
[0629] To a solution of 1H-imidazole (42 g, 617 mmol) in
dichloromethane (1 L) was added cyanogen bromide (22.5, 212 mmol)
and the mixture was heated to reflux for 30 minutes, allowed to
cool to room temperature and the white solid was filtered off. The
filtrate was concentrated to 100 ml and stored in the refrigerator
for 3 days. The precipitated solid was filtered off to obtain 8 g
di(1H-imidazol-1-yl)methanimine (49.6 mmol, 8%). .sup.1H NMR (500
MHz, DMSO) .delta. ppm 8.09 (s, 1H), 7.55 (s, 1H), 7.13 (s,
1H).
Step B: [1,2,4]Triazolo[4,3-a]pyridine-3-amine
##STR00863##
[0631] To a solution of 2-hydrazinylpyridine (5.2 g, 47.6 mmol) in
THF (70 ml) was added di(1H-imidazol-1-yl)methanimine (7.8 g, 48.4
mmol). The mixture was heated to reflux overnight. The crude
mixture was evaporated and purified on a Biotage silica gel column
(0-25%, methanol-methylene chloride) collecting the purple-colored
spot, [1,2,4]triazolo[4,3-a]pyridine-3-amine (4.7 g, 35 mmol,
73.5%). .sup.1H NMR (500 MHz, DMSO) .delta. ppm 8.05-8.0 (m, 1H),
7.44-7.40 (m, 1H), 7.08-7.0 (m, 1H), 6.74-6.70 (m, 1H), 6.35 (s,
2H). MS (LCMS) [M+H]=134.98; R.T.=0.1 min.
Step C:
Dimethyl[1,2,4]-triazol[4,3-a]pyridine-3-ylcarbonimidodithioate
##STR00864##
[0633] To a solution of [1,2,4]triazolo[4,3-a]pyridine-3-amine (300
mg, 2.24 mmol) in DMF (5 ml) was added NaOH (20M, 0.25 ml),
CS.sub.2 (0.5 ml), NaOH (20M, 0.25 ml) and iodomethane (0.5 ml)
slowly over 10 minutes. The mixture was stirred at room temperature
for 1 h and 10 ml water was added to the reaction mixture. The
precipitated solid was filtered off, washed with water (100 ml),
and dried to obtain 230 mg dimethyl[1, 2,
4]triazol[4,3-a]pyridine-3-ylcarbonimidodithioate (0.96 mmol,
43.1%), a white solid. .sup.1H NMR (500 MHz, CDCl3) .delta. ppm
8.17 (d, 1H), 7.7 (d, 1H), 7.24-7.22 (t, 1H), 6.84-6.80 (t, 1H),
2.71-2.68 (d, 6H). MS (LCMS) [M+H]=238.94; R.T.=1.26 min.
Step D:
(R)--N-([1,2,4]Triazol[4,3-a]pyridine-3-yl)-4H-1'-azaspiro[oxazole-
-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00865##
[0635] A mixture of
dimethyl[1,2,4]triazol[4,3-a]pyridine-3-ylcarbonimidothioate (120
mg, 0.50 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(120 mg, 0.76 mmol) and cesium carbonate (492 mg, 1.5 mmol) in DMF
(5 ml) was heated at 70.degree. C. for 6 hours. The mixture was
concentrated and purified on a Biotage silica gel column (100%
ethyl acetate, then 10-35% 9:1 methanol:ammonium
hydroxide-chloroform) to obtain the desired product,
(R)--N-([1,2,4]triazol[4,3-a]pyridine-3-yl)-4H-1'-azaspiro[oxazo-
le-5,3'-bicyclo[2.2.2]octan]-2-amine (97.2 mg, 61.5%). .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 8.2-8.1 (d, 1H), 7.6-7.5 (d, 1H),
7.2-7.1 (t, 1H), 6.7-6.6 (t, 1H), 4.1-4.0 (d, 1H), 3.7-3.6 (d, 1H),
3.5-3.4 (m, 1H), 3.1-2.7 (m, 5H), 2.4-2.2 (m, 2H), 1.8-1.7 (m, 1H),
1.7-1.5 (m, 2H). MS (LCMS) [M+H]=299.3; R.T.=1.22 min.
Example 263
(R)--N-(6-bromothiazolo[5,4-b]pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine
##STR00866##
[0636] Step A: Dimethyl
6-bromothiazolo[5,4-b]pyrazin-2-ylcarbonimidodithioate
##STR00867##
[0638] To a suspension of 6-bromothiazolo[5,4-b]pyrazin-2-amine
(700 mg, 3.03 mmol) in DMF (3 mL) was added 16.0M sodium hydroxide
(400 mL, 6.40 mmol). The mixture was allowed to stir 10 min at room
temperature at which time carbon disulfide was added (450 .mu.L,
7.57 mmol) and the resulting reddish brown mixture was stirred for
10 minutes. An additional portion of 16.0M sodium hydroxide (400
.mu.L, 6.40 mmol) was added and the mixture was again stirred for
10 minutes. Finally, iodomethane (450 .mu.L, 7.27 mmol) was added
dropwise. The mixture was stirred for 5 minutes, at which time a
voluminous yellow precipitate had formed. The mixture was poured
into water and the solids were collected by filtration to afford
dimethyl 6-bromothiazolo[5,4-b]pyrazin-2-ylcarbonimidodithioate
(680 mg, 67% yield) as a yellow solid of sufficient purity to use
without further purification. 1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 8.63 (s, 1H) 2.68 (s, 6H).
Step B:
(R)--N-(6-bromothiazolo[5,4-b]pyrazin-2-yl)-4H-1'-azaspiro[oxazole-
-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00868##
[0640] A mixture of dimethyl
6-bromothiazolo[5,4-b]pyrazin-2-ylcarbonimidodithioate (300 mg,
0.895 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(210 mg, 0.895 mmol) and cesium carbonate (600 mg, 1.79 mmol) in
acetonitrile (25 mL) was heated on a 100.degree. C. oil bath for 2
hours in an open flask, with nitrogen bubbling through the solution
the entire time to help in the removal of methanethiol. After 2
hours, TLC showed the reaction to be complete, so the mixture was
cooled to ambient temperature, diluted with water and concentrated
in vacuo. The mixture was extracted with chloroform (4.times.). The
combined organics were washed with brine, dried over sodium
sulfate, filtered, concentrated in vacuo, and the crude residue was
purified by silica gel chromatography (2-40% 9:1 methanol:ammonium
hydroxide-chloroform). The product fractions were combined and
concentrated in vacuo to afford
(R)--N-(6-bromothiazolo[5,4-b]pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine (200 mg, 57% yield).
[0641] 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.39 (br. s., 1H)
8.48 (s, 1H) 4.05 (d, J=9.79 Hz, 1H) 3.72 (d, J=9.79 Hz, 1H) 3.42
(dd, J=15.06, 1.76 Hz, 1H) 2.73-3.08 (m, 5 H) 2.10-2.22 (m, 2H)
1.73-1.84 (m, J=14.09, 9.94, 4.17, 4.17 Hz, 1H) 1.52-1.65 (m, 2H).
MS (LC/MS) R.T.=1.29; [M+H].sup.+=394.99.
Example 264
(R)--N-(6-(methylthio)thiazolo[5,4-b]pyrazin-2-yl)-4H-1'-azaspiro[oxazole--
5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00869##
[0643] A mixture of dimethyl
6-bromothiazolo[5,4-b]pyrazin-2-ylcarbonimidodithioate from Step A
of Example 263 (100 mg, 0.298 mmol),
(S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (68 mg, 0.298
mmol) and cesium carbonate (100 mg, 0.60 mmol) in DMF (1.5 mL) was
placed in a 1 dram vial and heated on a 100.degree. C. oil bath for
1 hour, at which time, sodium thiomethoxide (100 mg, 1.43 mmol) was
added and the mixture was heated overnight. The mixture was cooled
to ambient temperature and poured into water (20 mL) and the
resulting solids were collected by filtration and then purified by
silica gel chromatography (2-40% 9:1 methanol:ammonium
hydroxide-chloroform). The product fractions were combined and
concentrated in vacuo to afford
(R)--N-(6-(methylthio)thiazolo[5,4-b]pyrazin-2-yl)-4H-1'-azaspiro[oxazole-
-5,3'-bicyclo[2.2.2]octan]-2-amine (52 mg, 46% yield). 1H NMR (500
MHz, CDCl.sub.3) .delta. ppm 9.39 (br. s., 1H) 8.31 (s, 1H) 4.03
(d, J=9.77 Hz, 1H) 3.70 (d, J=9.77 Hz, 1H) 3.41 (dd, J=14.95, 1.83
Hz, 1H) 2.73-3.10 (m, 5H) 2.63 (s, 3H) 2.10-2.25 (m, 2H) 1.47-1.86
(m, 3H). MS (LC/MS) R.T.=1.04; [M+H].sup.+=363.04.
Example 265
(R)--N-(5-Methoxythiazolo[5,4-d]pyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine
##STR00870##
[0644] Step A: 5-Methoxythiazolo[5,4-d]pyrimidin-2-amine
##STR00871##
[0646] Ethyl 5-chlorothiazolo[5,4-d]pyrimidin-2-ylcarbamate (250
mg, 0.966 mmol) was suspended in MeOH (10 mL) and a 25% (w/w)
solution of sodium methoxide in methanol was added (10 mL, 46.3
mmol). The resulting solution was refluxed overnight, cooled to
ambient temperature, poured into an equal volume of water and
extracted with chloroform (4.times.). A significant amount of
compound was still present in the aqueous phase, so this was
concentrated to residue, and then dissolved in a small amount of 1N
HCl (not enough to make the resulting solution acidic) and
extracted again with EtOAc (5.times.). The combined organics were
washed with brine, dried over sodium sulfate, filtered and
concentrated in vacuo. 5-Methoxythiazolo[5,4-d]pyrimidin-2-amine
(144 mg, 0.790 mmol, 82% yield) was thus obtained as a white solid.
1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 8.43 (s, 1H) 7.81 (s,
2H) 3.90 (s, 3H). MS (LC/MS) R.T.=0.73; [M+H].sup.+=183.03.
Step B: Dimethyl
5-methoxythiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate
##STR00872##
[0648] To a suspension of 5-methoxythiazolo[5,4-d]pyrimidin-2-amine
(911 mg, 5.00 mmol) in DMF (5 mL) was added 20.0M sodium hydroxide
(500 .mu.L, 10.00 mmol). The mixture was allowed to stir 10 min at
room temperature at which time carbon disulfide was added (750
.mu.L, 12.50 mmol) and the resulting reddish brown mixture was
stirred for 10 minutes. An additional portion of 20.0M sodium
hydroxide (500 .mu.L, 10.00 mmol) was added and the mixture was
again stirred for 10 minutes. Finally, iodomethane (750 .mu.L,
12.00 mmol) was added dropwise. The mixture was stirred for 5
minutes, at which time a voluminous yellow precipitate had formed.
The mixture was poured into water and the solids were collected by
filtration to afford a yellow solid that was further purified by
silica gel chromatography (2-20% EtOAc/CHCl.sub.3) to provide
dimethyl 5-methoxythiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate
(380 mg, 27% yield).
[0649] 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.90 (s, 1H) 4.09
(s, 3H) 2.66 (s, 6H).
Step C:
(R)--N-(5-methoxythiazolo[5,4-d]pyrimidin-2-yl)-4H-1'-azaspiro[oxa-
zole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00873##
[0651] A mixture of dimethyl
5-methoxythiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate (100 mg,
0.349 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(80 mg, 0.349 mmol) and cesium carbonate (228 mg, 0.698 mmol) in
DMF (1.7 mL) was heated to 100.degree. C. for 2 hours. The reaction
mixture was cooled to ambient temperature, poured into water and
the solids were collected by filtration to afford
(R)--N-(5-methoxythiazolo[5,4-d]pyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine (78 mg, 64% yield). 1H NMR (500
MHz, CDCl.sub.3) .delta. ppm 9.12 (br. s., 1H) 8.63 (s, 1H)
3.95-4.18 (m, 4H) 3.71 (d, J=9.77 Hz, 1H) 3.41 (d, J=15.26 Hz, 1H)
2.74-3.10 (m, 5H) 2.11-2.27 (m, 2H) 1.71-1.86 (m, 1H) 1.50-1.70 (m,
2H). MS (LC/MS) R.T.=1.66; [M+H].sup.+=347.0.
Example 266
(R)--N-(5-Ethyl-1,3,4-oxadiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine
##STR00874##
[0652] Step A: Dimethyl
5-ethyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate
##STR00875##
[0654] To a solution of 5-ethyl-1,3,4-oxadiazol-2-amine (2.26 g, 20
mmol) in DMF (10 ml) was added NaOH (20M, 2 ml), CS.sub.2 (3 ml),
NaOH (20M, 2 ml) and iodomethane (3 ml) slowly over 10 minutes. The
mixture was stirred at room temperature for 2 h and poured into 30
ml water. The precipitated yellow solid was filtered off, washed
with water, and dried to obtain the desired product, dimethyl
5-ethyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate, a white solid
(2.6 g, 59.8%). .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm
2.86-2.83 (q, 2H), 2.63 (s, 6H), 1.3901.35 (t, 3H). MS (LCMS)
[M+H]=217.95; R.T.=1.93 min.
Step B:
(R)--N-(5-Ethyl-1,3,4-oxadiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine
##STR00876##
[0656] A mixture of dimethyl
5-ethyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate (327 mg, 1.5
mmol), (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (235 mg,
1.5 mmol) and cesium carbonate (1000 mg, 3.16 mmol) in DMF (10 ml)
was stirred overnight at room temperature. The mixture was
concentrated and purified on a Biotage silica gel column (100%
ethyl acetate, then 10-35% 9:1 methanol:ammonium
hydroxide-chloroform) to obtain the desired product,
(R)--N-(5-ethyl-1,3,4-oxadiazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine (290 mg, 66%). .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 4.05 (d, 1H), 3.74 (d, 1H), 3.3-3.2 (d, 1H), 3.2-3.1
(d, 1H), 3.0-2.9 (m, 2H), 2.9-2.8 (m, 5H), 2.2 (s, 1H), 2.15-2.0
(m, 1H), 1.9-1.6 (m, 3H), 1.4-1.3 (t, 3H). (m, 2H). MS (LCMS)
[M+H]=278.09; R.T.=0.48 min.
Example 267
(R)--N-(3,5-Dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00877##
[0657] Step A: 3,5-Dichloro-2-isothiocyanatopyridine
##STR00878##
[0659] To 3,5-dichloropyridin-2-amine (0.36 g, 2.209 mmol) in
dichloromethane (25 mL) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (0.523 g, 2.253 mmol). The
reaction was stirred at 40.degree. C. for 3 hours. The reaction was
cooled to room temperature and the crude was purified by
chromatography (Biotage: 25-100% ethyl acetate/hexane) to yield to
yield 3,5-dichloro-2-isothiocyanatopyridine (0.4 g, 1.951 mmol, 88%
yield). 1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.50 (t, J=2.59 Hz, 1
H), 8.45 (t, J=2.59 Hz, 1H). MS (LC/MS) R.T.=2.07;
[M+H].sup.+=204.8.
Step B:
(R)--N-(3,5-Dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00879##
[0661] To 3,5-dichloro-2-isothiocyanatopyridine (0.11 g, 0.55 mmol)
in N,N-dimethylformamide (10 mL) was added Et.sub.3N (0.17 mL, 1.21
mmol) and (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.13
g, 0.56 mmol) at room temperature. The reaction was stirred at
70.degree. C. for 2 hours. The reaction was cooled to room
temperature and concentrated in vacuo. The crude urea was purified
by chromatography (Biotage: 85% CHCl.sub.3, 14% MeOH, 1%
NH.sub.4OH). The product was then treated with
N,N-dimethylformamide (10 mL) and N,N'-diisopropylcarbodiimide
(0.26 mL, 1.65 mmol). The reaction was heated to 70.degree. C. for
2 hours. The reaction was cooled to room temperature and
concentrated in vacuo to yield the crude product. The crude product
was purified by chromatography (Biotage: 85% CHCl.sub.3, 14% MeOH,
1% NH.sub.4OH) to yield
(R)--N-(3,5-dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2-
.2]octan]-2-amine (0.08 g, 0.24 mmol, 44% yield) as a white powder.
1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.91 (s, 1H), 8.11-8.17 (m,
1H), 7.97 (d, J=2.44 Hz, 1H), 3.84 (d, J=9.77 Hz, 1H), 3.59 (d,
J=9.77 Hz, 1H), 2.95-3.04 (m, 2H), 2.72-2.81 (m, 2H), 2.66 (t,
J=7.63 Hz, 2H), 2.01 (s, 1H), 1.89 (s, 1H), 1.54-1.62 (m, 2H),
1.42-1.50 (m, 1 H). MS (LC/MS) R.T.=0.78; [M+].sup.+=326.1.
Example 268
(R)--N-(5-chlorothiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'
bicyclo[2.2.2]octan]-2-amine
##STR00880##
[0662] Step A: Dimethyl
5-chlorothiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate
##STR00881##
[0664] To a suspension of 5-chlorothiazolo[5,4-b]pyridin-2-amine
(930 mg, 5.00 mmol) in DMF (5 mL) was added 20.0M sodium hydroxide
(500 .mu.L, 10.00 mmol). The mixture was allowed to stir 10 min at
room temperature at which time carbon disulfide was added (750
.mu.L, 12.50 mmol) and the mixture was stirred for 10 minutes. An
additional portion of 20.0M sodium hydroxide (500 .mu.L, 10.0 mmol)
was added and the mixture was again stirred for 10 minutes.
Finally, iodomethane (750 .mu.L, 12.00 mmol) was added dropwise. An
exotherm was noticed during this addition. The mixture was stirred
for 15 minutes, at which time a voluminous precipitate had formed.
The mixture was poured into water and the solids were collected by
filtration. Most of the collected solids were pale yellow and
crystalline. A few small clumps of a slightly darker gummy orange
solid were also present, and these were manually removed and
discarded. The remainder was the title compound, dimethyl
5-chlorothiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate (1.00 g,
69% yield). 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.04 (d,
J=8.53 Hz, 1H) 7.38 (d, J=8.53 Hz, 1H) 2.66 (s, 6H).
Step B:
(R)--N-(5-chlorothiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazol-
e-5,3' bicyclo[2.2.2]octan]-2-amine
##STR00882##
[0666] A mixture of dimethyl
5-chlorothiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate (100 mg,
0.35 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (79
mg, 0.35 mmol) and cesium carbonate (225 mg, 0.69 mmol) in DMF (1.7
mL) was heated to 100.degree. C. for 2 hours. The reaction mixture
was cooled to ambient temperature, poured into water and the solids
collected by filtration. The crude solids were purified by silica
gel chromatography (2-40% 9:1 methanol:ammonium
hydroxide-chloroform) to afford
(R)--N-(5-chlorothiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'b-
icyclo[2.2.2]octan]-2-amine (62 mg, 51% yield). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.13 (br. s., 1H) 7.93 (d, J=8.53 Hz, 1H)
7.45 (d, J=8.28 Hz, 1H) 3.92 (d, J=10.29 Hz, 1H) 3.67 (d, J=10.29
Hz, 1H) 3.00-3.14 (m, 2H) 2.77-2.93 (m, 2H) 2.69 (t, J=7.65 Hz, 2H)
2.12 (br. s., 1H) 1.95 (br. s., 1H) 1.43-1.72 (m, 3H). MS (LC/MS)
R.T.=1.10; [M+H].sup.+=350.10.
Example 269
(R)--N.sup.5,N.sup.5-dimethyl-N.sup.2-(4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octane]-2-yl)thiazolo[5,4-d]pyrimidine-2,5-diamine
##STR00883##
[0667] Step A: Ethyl
5-chlorothiazolo[5,4-d]pyrimidin-2-ylcarbamate
##STR00884##
[0669] Ethoxycarbonyl isothiocyanate (4.32 mL, 36.6 mmol) and
2,4-dichloro-pyrimidin-5-ylamine (3.00 g, 18.29 mmol) were mixed
neat and sonicated for 5 minutes to help dissolve. The mixture was
stirred at ambient temperature for 10 minutes, at which time the
entire mixture had solidified. Methanol (100 mL) was added and the
mixture was refluxed for 30 minutes, cooled to ambient temperature
and the solids were collected by filtration to afford ethyl
5-chlorothiazolo[5,4-d]pyrimidin-2-ylcarbamate (3.8 g, 80% yield).
1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.64 (s, 1H) 9.05 (s,
1H) 4.31 (q, J=7.19 Hz, 2H) 1.32 (t, J=7.15 Hz, 3H).
Step B: Ethyl
5-(dimethylamino)thiazolo[5,4-d]pyrimidin-2-ylcarbamate
##STR00885##
[0671] Ethyl 5-chlorothiazolo[5,4-d]pyrimidin-2-ylcarbamate (300
mg, 1.16 mmol) was suspended in a 2.0 M solution of dimethylamine
in methanol (5.0 mL, 10.00 mmol) in a pressure vessel, which was
sealed and heated overnight on a 75.degree. C. oil bath. The
mixture was cooled to ambient temperature, the solvent was
evaporated and the residue was partitioned between aqueous
bicarbonate and chloroform and extracted 3 times. The combined
organics were washed with brine, dried over sodium sulfate,
filtered and concentrated in vacuo to afford ethyl
5-(dimethylamino)thiazolo[5,4-d]pyrimidin-2-ylcarbamate (236 mg,
99% yield). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 11.97 (s,
1H) 8.67 (s, 1H) 4.26 (q, J=7.03 Hz, 2 H) 3.17 (s, 6H) 1.16-1.40
(m, 3H). MS (LC/MS) R.T.=1.88; [M+H].sup.+=268.09.
Step C: Ethyl
N.sup.5,N.sup.5-dimethylthiazolo[5,4-d]pyrimidine-2,5-diamine
##STR00886##
[0673] Ethyl
5-(dimethylamino)thiazolo[5,4-d]pyrimidin-2-ylcarbamate (236 mg,
0.88 mmol) was suspended in a 25% (w/w) solution of sodium
methoxide in methanol (5 mL, 23.0 mmol) and the mixture was heated
to reflux overnight. The reaction mixture was evaporated to dryness
and the residue was partitioned between water and chloroform and
extracted 3 times. The combined organics were washed with brine,
dried over sodium sulfate, filtered and concentrated in vacuo to
afford ethyl
N.sup.5,N.sup.5-dimethylthiazolo[5,4-d]pyrimidine-2,5-diamine (170
mg, 99% yield) as a white solid. 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 8.27 (s, 1H) 7.44 (s, 2H) 3.10 (s, 6H).
Step D: Dimethyl
5-(dimethylamino)thiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate
##STR00887##
[0675] To a suspension of ethyl
N.sup.5,N.sup.5-dimethylthiazolo[5,4-d]pyrimidine-2,5-diamine (160
mg, 0.819 mmol) in DMF (1 mL) was added 20.0 M sodium hydroxide
(100 .mu.L, 2.00 mmol). The mixture was allowed to stir 10 min at
room temperature at which time carbon disulfide was added (120
.mu.L, 2 mmol) and the resulting reddish brown mixture was stirred
for 10 minutes. An additional portion of 20.0 M sodium hydroxide
(100 .mu.L, 2.0 mmol) was added and the mixture was again stirred
for 10 minutes. Finally, iodomethane (120 .mu.L, 1.9 mmol) was
added dropwise. The mixture was stirred for 5 minutes, at which
time a voluminous yellow precipitate had formed. The mixture was
poured into water and the solids were collected by filtration to
afford dimethyl
5-(dimethylamino)thiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate
(194 mg, 79% yield). 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.75
(s, 1H) 3.26 (s, 6H) 2.64 (s, 6H).
Step E:
(R)--N.sup.5,N.sup.5-dimethyl-N.sup.2-(4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octane]-2-yl)thiazolo[5,4-d]pyrimidine-2,5-diamine
##STR00888##
[0677] A mixture of dimethyl
5-(dimethylamino)thiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate
(90 mg, 0.301 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol
dihydrochloride (83 mg, 0.361 mmol) and cesium carbonate (196 mg,
0.60 mmol) in DMF (1.0 mL) was heated to 100.degree. C. for 1.5
hours. The reaction mixture was cooled to ambient temperature,
poured into water and extracted with chloroform (4.times.). The
combined organics were washed with brine, dried over sodium
sulfate, filtered, concentrated in vacuo, and the crude residue was
purified by silica gel chromatography (2-40% 9:1 methanol:ammonium
hydroxide-chloroform) to afford
(R)--N.sup.5,N.sup.5-dimethyl-N.sup.2-(4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octane]-2-yl)thiazolo[5,4-d]pyrimidine-2,5-diamine (81 mg,
71% yield) as a tan solid. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 9.07 (br. s., 1H) 8.50 (s, 1H) 4.01 (d, J=9.54 Hz, 1H)
3.67 (d, J=9.54 Hz, 1H) 3.39 (dd, J=14.93, 1.63 Hz, 1H) 3.23 (s,
6H) 2.71-3.10 (m, 5H) 2.10-2.24 (m, 2H) 1.68-1.84 (m, 1H) 1.46-1.68
(m, 2H). MS (LC/MS) R.T.=0.87; [M+H].sup.+=360.23.
Example 270
(R)--N-([1,2,4]Triazol[1,5-a]pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00889##
[0678] Step A: [1,2,4]-Triazolo[1,5-a]pyrazin-2-amine
##STR00890##
[0680] To a solution of pyrazin-2-amine (25 g, 260 mmol) in dioxane
(300 ml) at room temperature was added
ethoxycarbonyl-isothiocyanate (37.9 g, 289 mmol) slowly. The
mixture was stirred for 18 hours and the solvent was evaporated
under vacuum. The residual solid was dissolved in a mixture of
methanol (150 ml) and ethanol (150 ml). To this solution was added
TEA (109 ml, 780 mmol) and hydroxylamine hydrochloride (72.5 g,
1040 mmol). The mixture was stirred at room temperature for 2 hours
and was heated to reflux for 4 hours. The crude mixture was cooled
to room temperature and the solvent was evaporated. The residual
solid was purified by column chromatography (0-20%
methanol/CH.sub.2Cl.sub.2) to obtain a white solid (60 g). The
solid was taken into EtOAc and water. The aqueous layer was
extracted with EtOAc twice. The combined organic layer was washed
with brine and dried over sodium sulfate to obtain
[1,2,4]triazolo[1,5-a]pyrazin-2-amine as a white solid (12 g, 88
mmol, 33%). MS (LCMS) [M+H]=135.96; R.T.=0.21 min.
Step B:
Dimethyl[1,2,4]triazol[1,5-a]pyrazin-2-ylcarbonimidodithioate
##STR00891##
[0682] To a solution of [1,2,4]triazolo[1,5-a]pyrazin-2-amine (676
mg, 5 mmol) in DMF (10 ml) was added NaOH (20 M, 0.5 ml), CS.sub.2
(1 ml), NaOH (20 M, 0.5 ml) and iodomethane (1 ml) slowly over 10
minutes. The mixture was stirred at room temperature for 1 h and 10
ml water was added to the reaction mixture, which became cloudy.
The mixture was extracted with EtOAc (100 ml.times.3). The combined
organic layers were washed with brine, dried over sodium sulfate,
filtered, and evaporated. The residue was purified on a Biotage
silica gel column (ethyl acetate-hexane 10-30%) to obtain
dimethyl[1,2,4]triazol[1,5-a]pyrazin-2-ylcarbonimidodithioate as a
yellow solid (720 mg, 3 mmol, 60%). .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 9.2 (2, 1H), 8.5 (d, 1H), 8.2 (d, 1H), 2.67
(s, 6H). MS (LCMS) [M+H]=239.92. [M+Na]=261.89; R.T.=1.55 min.
Step C:
(R)--N-([1,2,4]Triazol[1,5-a]pyrazin-2-yl)-4H-1'-azaspiro[oxazole--
5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00892##
[0684] A mixture of
dimethyl[1,2,4]triazol[1,5-a]pyrazin-2-ylcarbonimidodithioate (120
mg, 0.50 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(120 mg, 0.76 mmol) and cesium carbonate (492 mg, 1.5 mmol) in DMF
(5 ml) was heated at 70.degree. C. for 6 hours. The mixture was
concentrated and purified on a Biotage silica gel column (100%
ethyl acetate, then 10-35% 9:1 methanol:ammonium
hydroxide-chloroform) to obtain the desired product,
(R)--N-([1,2,4]triazol[1,5-a]pyrazin-2-yl)-4H-1'-azaspiro[oxazol-
e-5,3'-bicyclo[2.2.2]octan]-2-amine (85 mg, 26.7%) as a white
solid. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 9.0 (m, 1H),
8.71-8.70 (m, 1H), 8.15-8.10 (m, 1H), 4.15-4.0 (d, 1H), 3.85-3.8
(d, 1H), 3.6-3.5 (d, 1H), 3.4-3.3 (d, 1H), 3.3-3.0 (m, 4H), 2.4-2.2
(m, 2H), 2.0-1.8 (m, 3H).
[0685] MS (LCMS) [M+H]=300.06; R.T.=0.2 min.
Example 271
(R)--N-(Thiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine
##STR00893##
[0686] Step A: Dimethyl
thiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate
##STR00894##
[0688] To a suspension of thiazolo[5,4-b]pyridin-2-amine (300 mg,
1.98 mmol) in DMF (2 mL) was added 20.0M sodium hydroxide (200
.mu.L, 4.0 mmol). The mixture was allowed to stir 10 min at room
temperature at which time carbon disulfide was added (300 .mu.L,
4.96 mmol) and the resulting reddish brown mixture was stirred for
10 minutes. An additional portion of 20.0M sodium hydroxide (200
.mu.L, 4.0 mmol) was added and the mixture was again stirred for 10
minutes. Finally, iodomethane (300 .mu.L, 4.76 mmol) was added
dropwise. The mixture was stirred for 5 minutes, at which time a
voluminous yellow precipitate had formed. The mixture was poured
into water and the solids were collected by filtration to afford
dimethyl thiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate (190 mg,
38% yield) as a yellow solid of sufficient purity to use without
further purification. 1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 8.47
(d, J=4.58 Hz, 1H) 8.11 (dd, J=8.24, 1.53 Hz, 1H) 7.37 (dd, J=8.24,
4.88 Hz, 1H) 2.66 (s, 6H).
Step B:
(R)--N-(Thiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine
##STR00895##
[0690] A mixture of dimethyl
thiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate (90 mg, 0.35 mmol),
(S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (97 mg, 0.42
mmol) and cesium carbonate (230 mg, 0.71 mmol) in DMF (1 mL) was
heated to 100.degree. C. for 2 hours. The reaction mixture was
cooled to ambient temperature, poured into water and extracted with
chloroform (4.times.). The combined organics were washed with
brine, dried over sodium sulfate, filtered and concentrated in
vacuo. The mixture was purified by silica gel chromatography (2-40%
9:1 methanol:ammonium hydroxide-chloroform). The product fractions
were combined and concentrated in vacuo to afford
(R)--N-(thiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine (84 mg, 76% yield). 1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 9.30 (br. s., 1H) 8.37 (dd, J=4.77, 1.51
Hz, 1H) 7.82 (dd, J=8.03, 1.51 Hz, 1H) 7.28 (dd, J=8.03, 4.77 Hz,
1H) 4.05 (d, J=9.54 Hz, 1H) 3.70 (d, J=9.54 Hz, 1H) 3.42 (dd,
J=15.06, 1.76 Hz, 1H) 2.75-3.07 (m, 5H) 2.14-2.26 (m, 2H) 1.71-1.84
(m, J=13.99, 9.79, 4.17, 4.17 Hz, 1H) 1.48-1.68 (m, 2H). MS (LC/MS)
R.T.=0.64; [M+H].sup.+=316.15.
Example 272
(R)--N-(Thiazolo[5,4-b]pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine
##STR00896##
[0692]
(R)--N-(6-Bromothiazolo[5,4-b]pyrazin-2-yl)-4H-1'-azaspiro[oxazole--
5,3'-bicyclo[2.2.2]octan]-2-amine (44 mg, 0.111 mmol) was suspended
in MeOH (50 mL) and 3N HCl was added until all solids had dissolved
(-10 ml). The reaction flask was flushed with nitrogen and then 10%
palladium on carbon (35 mg) was added, and the flask was fitted
with a hydrogen balloon. The mixture was allowed to react
overnight, at which time TLC showed consumption of the starting
material. The flask was flushed with nitrogen, filtered through
celite and washed with methanol. The combined filtrates were
concentrated by .about.90% to remove most of the methanol, and the
solution was made basic by the addition of a saturated solution of
sodium bicarbonate. The basic aqueous phase was extracted with
chloroform (4.times.). The combined organics were washed with
brine, dried over sodium sulfate, filtered and concentrated in
vacuo. The mixture was purified by silica gel chromatography (2-40%
[9:1 methanol:ammonium hydroxide]-chloroform). The product
fractions were combined and concentrated in vacuo to afford
(R)--N-(thiazolo[5,4-b]pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine (24 mg, 0.075 mmol, 67.5% yield). 1H NMR (400
MHz, CDCl.sub.3) .delta. ppm 9.50 (br. s., 1H) 8.41 (d, J=2.76 Hz,
1 H) 8.27 (d, J=2.76 Hz, 1H) 4.06 (d, J=9.79 Hz, 1H) 3.72 (d,
J=9.79 Hz, 1H) 3.43 (dd, J=15.06, 1.76 Hz, 1H) 2.74-3.09 (m, 5H)
2.12-2.25 (m, 2H) 1.71-1.86 (m, 1H) 1.49-1.67 (m, 2H). MS (LC/MS)
R.T.=0.75; [M+H].sup.+=317.13.
Example 273
(R)--N-(7-Methoxy-5-methylthiazolo[5,4-d]pyrimidin-2-yl)-4H-1'-azaspiro[ox-
azole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00897##
[0693] Step A: Ethyl
7-chloro-5-methylthiazolo[5,4-d]pyrimidin-2-ylcarbamate
##STR00898##
[0695] To a mixture of 4,6-dichloro-2-methylpyrimidin-5-amine (1 g,
5.62 mmol) and O-ethyl carbonisothiocyanatidate (0.66 mL, 5.62
mmol) was added toluene (2 mL) to wet the solids completely. The
mixture was placed on 100.degree. C. oil bath for 1.5 hours, at
which time, the mixture had seized to give a solid mass. The solids
were cooled to ambient temperature and triturated with ether, then
the resulting solids were collected by filtration to give ethyl
7-chloro-5-methylthiazolo[5,4-d]pyrimidin-2-ylcarbamate (1.08 g,
3.96 mmol, 70.5% yield). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
12.70 (br. s., 1H) 4.30 (q, J=7.19 Hz, 2H) 2.69 (s, 3H) 1.30 (t,
J=7.15 Hz, 3 H).
Step B: 7-Methoxy-5-methylthiazolo[5,4-d]pyrimidin-2-amine
##STR00899##
[0697] Ethyl
7-chloro-5-methylthiazolo[5,4-d]pyrimidin-2-ylcarbamate (300 mg,
1.100 mmol) was suspended in a 25% w/w solution of sodium methoxide
in methanol (5 mL, 23.14 mmol) and the mixture was refluxed
overnight. The mixture was cooled to ambient temperature, diluted
with water and extracted with chloroform (4.times.). The combined
organics were washed with brine, dried over sodium sulfate,
filtered and concentrated in vacuo to afford
7-methoxy-5-methylthiazolo[5,4-d]pyrimidin-2-amine (120 mg, 0.612
mmol, 55.6% yield). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.71
(s, 2H) 3.98 (s, 3H) 2.52 (s, 3H).
Step C: Dimethyl
7-methoxy-5-methylthiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate
##STR00900##
[0699] To a suspension of
7-methoxy-5-methylthiazolo[5,4-d]pyrimidin-2-amine (100 mg, 0.51
mmol) in DMF (0.5 mL) was added 16.0M sodium hydroxide (75 .mu.L,
1.2 mmol). The mixture was allowed to stir 10 min at room
temperature at which time carbon disulfide was added (80 .mu.L,
1.27 mmol) and the resulting reddish brown mixture was stirred for
10 minutes. An additional portion of 16.0M sodium hydroxide (75
.mu.L, 1.2 mmol) was added and the mixture was again stirred for 10
minutes. Finally, iodomethane (80 .mu.L, 1.29 mmol) was added
dropwise. The mixture was stirred for 5 minutes, at which time a
voluminous yellow precipitate had formed. The mixture was poured
into water and the solids were collected by filtration to afford a
crude yellow solid that was further purified by silica gel
chromatography (2-20% ethyl acetate-chloroform). The product
fractions were combined and concentrated in vacuo to afford
dimethyl
7-methoxy-5-methylthiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate
(90 mg, 59% yield) as a yellow solid. 1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 4.17 (s, 3H) 2.71 (s, 3H) 2.64 (s, 6 H).
Step D:
(R)--N-(7-Methoxy-5-methylthiazolo[5,4-d]pyrimidin-2-yl)-4H-1'-aza-
spiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00901##
[0701] A mixture of dimethyl
7-methoxy-5-methylthiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate
(56 mg, 0.19 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol
dihydrochloride (51 mg, 0.22 mmol) and cesium carbonate (175 mg,
0.54 mmol) in DMF (0.5 mL) was heated to 100.degree. C. for 2
hours. The reaction mixture was cooled to ambient temperature,
poured into water and extracted with chloroform (4.times.). The
combined organics were washed with brine, dried over sodium
sulfate, filtered and concentrated in vacuo. The mixture was
purified by silica gel chromatography (2-40% [9:1 methanol:ammonium
hydroxide]-chloroform). The product fractions were combined and
concentrated in vacuo to afford
(R)--N-(7-methoxy-5-methylthiazolo[5,4-d]pyrimidin-2-yl)-4H-1'-azaspiro[o-
xazole-5,3'-bicyclo[2.2.2]octan]-2-amine (34 mg, 50% yield). 1H NMR
(400 MHz, CDCl.sub.3) .delta. ppm 9.10 (br. s., 1H) 4.14 (s, 3H)
4.03 (d, J=9.54 Hz, 1H) 3.68 (d, J=9.54 Hz, 1H) 3.39 (dd, J=14.93,
1.63 Hz, 1H) 2.74-3.07 (m, 5H) 2.68 (s, 3H) 2.04-2.28 (m, 2 H)
1.70-1.86 (m, 1H) 1.44-1.67 (m, 2H). MS (LC/MS) R.T.=1.10;
[M+H].sup.+=361.32.
Example 274
(R)--N-(7-Methoxythiazolo[5,4-d]pyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine
##STR00902##
[0702] Step A: Dimethyl
7-methoxy-5-methylthiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate
##STR00903##
[0704] To a suspension of 7-methoxythiazolo[5,4-d]pyrimidin-2-amine
(300 mg, 1.67 mmol) in DMF (1.5 mL) was added 16.0M sodium
hydroxide (210 .mu.L, 3.4 mmol). The mixture was allowed to stir 10
min at room temperature at which time carbon disulfide was added
(250 .mu.L, 4.15 mmol) and the resulting reddish brown mixture was
stirred for 10 minutes. An additional portion of 16.0M sodium
hydroxide (210 .mu.L, 3.4 mmol) was added and the mixture was again
stirred for 10 minutes. Finally, iodomethane (250 .mu.L, 4.00 mmol)
was added dropwise. The mixture was stirred for 10 minutes, at
which time a voluminous yellow precipitate had formed. The mixture
was poured into water and the solids were collected by filtration
to afford dimethyl
7-methoxy-5-methylthiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate
(324 mg, 69% yield) as a yellow solid. 1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 8.60 (s, 1H) 4.20 (s, 3H) 2.65 (s, 6H).
Step B:
(R)--N-(7-Methoxythiazolo[5,4-d]pyrimidin-2-yl)-4H-1'-azaspiro[oxa-
zole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00904##
[0706] A mixture of dimethyl
7-methoxythiazolo[5,4-d]pyrimidin-2-ylcarbonimidodithioate (150 mg,
0.52 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(132 mg, 0.58 mmol) and cesium carbonate (427 mg, 1.31 mmol) in DMF
(3 mL) was heated to 100.degree. C. for 2 hours. The reaction
mixture was cooled to ambient temperature, poured into water and
extracted with chloroform (4.times.). The combined organics were
washed with brine, dried over sodium sulfate, filtered and
concentrated in vacuo. The mixture was purified by silica gel
chromatography (2-40% [9:1 methanol:ammonium
hydroxide]-chloroform). The product fractions were combined and
concentrated in vacuo to afford
(R)--N-(7-methoxythiazolo[5,4-d]pyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine (95 mg, 51% yield). 1H NMR (400
MHz, CDCl.sub.3) .delta. ppm 9.12 (br. s., 1H) 8.52 (s, 1H) 4.16
(s, 3H) 4.05 (d, J=9.54 Hz, 1H) 3.70 (d, J=9.54 Hz, 1H) 3.40 (dd,
J=14.93, 1.88 Hz, 1H) 2.70-3.07 (m, 5H) 2.08-2.27 (m, 2H) 1.68-1.85
(m, 1H) 1.48-1.66 (m, 2H). MS (LC/MS) R.T.=0.90;
[M+H].sup.+=347.34.
Example 275
(R)-2-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-ylamino)thiazole-
-5-carbonitrile
##STR00905##
[0708]
(R)-2-(4H-1'-Azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-ylamino)t-
hiazole-5-carbonitrile was synthesized by the method of Example
274, starting from 2-amino-5-cyanothiazole. 1H NMR (500 MHz,
DMSO-D6) .delta. ppm 9.05 (s, 1H), 8.13 (s, 1H), 3.86 (d, J=10.38
Hz, 1H), 3.61 (d, J=10.38 Hz, 1H), 3.01-3.10 (m, 2H), 2.83 (t,
J=7.63 Hz, 2H), 2.62-2.71 (m, 2H), 2.09 (s, 1H), 1.90-1.97 (m, 2H),
1.54-1.62 (m, 3H). MS (LC/MS) R.T.=0.52; [M+H].sup.+=290.0.
Example 276
(R)--N-(7-Bromopyrrolo[1,2-f][1,2,4]triazin-4-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR00906##
[0710]
(R)--N-(7-bromopyrrolo[1,2-f][1,2,4]triazin-4-yl)-4H-1'-azaspiro[ox-
azole-5,3'-bicyclo[2.2.2]octan]-2-amine was synthesized by the
method of Example 274 starting from
7-bromopyrrolo[1,2-f][1,2,4]triazin-4-amine 1H NMR (400 MHz, MeOD)
.delta. ppm 8.13 (1H, s), 7.04 (1H, d, J=4.53 Hz), 6.77 (1H, d,
J=4.53 Hz), 4.09 (1H, d, J=10.32 Hz), 3.79 (1H, d, J=10.58 Hz),
3.24 (1H, d), 3.12 (1H, d), 2.70-3.00 (4H, m), 2.06-2.25 (2H, m),
1.52-1.86 (3H, m) MS (LC/MS) R.T.=1.62; [M+H].sup.+=377.2.
Example 277
(R)--N-(1,6-Naphthyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]o-
ctan]-2-amine
##STR00907##
[0712]
(R)--N-(1,6-Naphthyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine was synthesized by the method of Example 274
starting from 1,6-naphthyridin-2-amine 1H NMR (400 MHz, MeOD)
.delta. ppm 8.99 (1H, s), 8.48 (1H, d, J=6.04 Hz), 8.20 (1H, d,
J=8.56 Hz), 7.77 (1H, d, J=6.04 Hz), 7.12 (1 H, d, J=8.81 Hz), 4.12
(1H, d, J=10.32 Hz), 3.82 (1H, d, J=10.32 Hz), 3.36 (1H, d), 3.21
(1H, d), 2.79-3.09 (4H, m), 2.08-2.30 (2H, m), 1.56-1.95 (3H, m).
(LC/MS) R.T.=0.38; [M+H].sup.+=310.3.
Example 278
(R)--N-(Quinazolin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]--
2-amine
##STR00908##
[0714]
(R)--N-(Quinazolin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]-
octan]-2-amine was synthesized by the method of Example 274
starting from 2-aminoquinazoline. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 9.27 (1H, s), 7.72-7.99 (3 H, m), 7.47 (1H, dd, J=7.55,
3.78 Hz), 4.07 (1H, d, J=10.07 Hz), 3.76 (1H, d, J=10.07 Hz), 3.26
(1H, br. s.), 3.13 (1H, d), 2.70-3.03 (4H, m), 2.17 (2H, br. s.),
1.50-1.88 (3H, m). (LC/MS) R.T.=1.11; [M+H].sup.+=310.3.
Example 279
(R)--N-(6,8-Dichloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine
##STR00909##
[0715] Step A: N-(2,4-Dichlorobenzyl)-2,2-diethoxyacetimidamide
##STR00910##
[0717] (2,4-Dichlorophenyl)methanamine (2 g, 11.4 mmol) was added
to a solution of methyl 2,2-diethoxyacetimidate (2.04 g, 12.6 mmol)
in methanol (10 ml). The mixture was heated at 70.degree. C. for 1
h. The mixture was purified by chromatography (Biotage: 100% ethyl
acetate). The desired fractions were concentrated to yield
N-(2,4-dichlorobenzyl)-2,2-diethoxyacetimidamide (2.8 g, 9.2 mmol,
72.7% yield) as a colorless viscous oil. 1H NMR (500 MHz,
DMSO-D.sub.6) .delta. ppm 7.27-7.70 (m, 3H), 4.77 (s, 1H),
4.14-4.35 (m, 2H), 3.45-3.68 (m, 4H), 1.09-1.29 (m, 6H). LC/MS
RT=2.03; [M+H].sup.+=304.9.
Step B: 6,8-Dichloroisoquinolin-3-amine
##STR00911##
[0719] To sulfuric acid (4 mL, 75 mmol) was added
N-(2,4-dichlorobenzyl)-2,2-diethoxyacetimidamide (2 g, 6.6 mmol) at
room temperature. The reaction was heated to 40.degree. C. for 18
hours. TLC and LC/MS indicated the presence of product. The
reaction was cooled to room temperature and quenched with aq. NaOH
(.about.15 M) until the reaction mixture was .about.pH 7. The crude
product was extracted with ethyl acetate (2.times.50 mL) and the
organics were dried with MgSO.sub.4, filtered, and concentrated in
vacuo to yield the product. The crude product was purified by
chromatography (Biotage: 10-80% ethyl acetate/hexanes) to yield
5,7-dichloroisoquinolin-1-amine (0.32 g, 1.50 mmol, 22.9% yield) as
a dark yellow powder. 1H NMR (400 MHz, DMSO-D.sub.6) .delta. ppm
8.99 (s, 1H), 7.64-7.73 (m, 1H), 7.30 (d, J=2.01 Hz, 1H), 6.61 (s,
1H), 6.43 (s, 2H). MS (LC/MS) R.T.=1.40; [M+H].sup.+=213.1.
Step C: 6,8-Dichloro-3-isothiocyanatoisoquinoline
##STR00912##
[0721] To 6,8-dichloroisoquinolin-3-amine (0.27 g, 1.28 mmol) in
dichloromethane (20 mL) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (0.30 g, 1.29 mmol) and the
reaction mixture was stirred at 40.degree. C. for 4 hours. The
reaction was cooled to room temperature and chromatographed
(Biotage: 10-100% ethyl acetate/hexanes) to yield
6,8-dichloro-3-isothiocyanatoisoquinoline (0.2 g, 0.78 mmol, 61.9%
yield) as a powder. 1H NMR (500 MHz, DMSO-D.sub.6) .delta. ppm 9.42
(s, 1H), 8.16 (d, J=1.83 Hz, 1 H), 8.02 (d, J=2.14 Hz, 1H), 7.92
(s, 1H). MS (LC/MS) R.T.=3.63; [M+H].sup.+=255.0.
Step D:
(R)--N-(6,8-Dichloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine
##STR00913##
[0723] To 6,8-dichloro-3-isothiocyanatoisoquinoline (0.17 g, 0.67
mmol) in DMF (10 mL) was added cesium carbonate (0.543 g, 1.67
mmol) and (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.15
g, 0.67 mmol) at room temperature. The reaction was stirred at
70.degree. C. for 2 hours. The reaction was cooled to room
temperature and concentrated in vacuo. The product was treated with
N,N'-diisopropylcarbodiimide (0.31 mL, 2.0 mmol). The reaction was
heated to 90.degree. C. for 4 hours. The reaction was cooled to
room temperature and concentrated to yield the crude product. The
crude product was purified by chromatography (Biotage: 85%
CHCl.sub.3, 14% MeOH, 1% NH.sub.4OH). The product was taken up in a
small amount of ethyl acetate, which resulted in the formation of a
precipate. It was filtered off, washed with a small amount of ethyl
acetate, and dried in a vacuum oven to yield
(R)--N-(6,8-dichloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine (0.094 g, 0.24 mmol, 36.6% yield) as a white
powder. 1H NMR (400 MHz, DMSO-D.sub.6) .delta. ppm 9.23 (s, 1H),
8.71-8.83 (m, 1H), 7.90-8.00 (m, 1H), 7.57-7.67 (m, 1H), 7.13-7.24
(m, 1H), 3.79-3.90 (m, 1H), 3.53-3.64 (m, 1H), 2.93-3.04 (m, 2H),
2.72-2.82 (m, 2H), 2.61-2.70 (m, 2H), 1.99 (s, 1H), 1.90 (s, 1H),
1.59 (d, J=4.78 Hz, 2H), 1.40-1.50 (m, 1H). MS (LC/MS) R.T.=1.68;
[M+H].sup.+=377.1.
Example 280
##STR00914##
[0724]
(R)--N-(6,7-Dichloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine
##STR00915##
[0725] Step A: N-(3,4-Dichlorobenzyl)-2,2-diethoxyacetimidamide
##STR00916##
[0727] (3,4-Dichlorophenyl)methanamine (2 g, 11.4 mmol) was added
to a solution of methyl 2,2-diethoxyacetimidate (2.04 g, 12.6 mmol)
in methanol (10 ml). The mixture was heated at 70.degree. C. for 1
h. The mixture was purified by chromatography (Biotage: 100% ethyl
acetate). The desired fractions were concentrated to yield
N-(3,4-dichlorobenzyl)-2,2-diethoxyacetimidamide (2.8 g, 9.2 mmol,
72.7% yield) as a colorless viscous oil. 1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 7.45 (m, 1H), 7.40 (m, 1 H), 7.19 (dd,
J=8.09, 1.98 Hz, 1H), 4.94 (s, 1H), 4.43 (s, 2H), 3.47-3.77 (m,
4H), 1.41-1.79 (m, 6H). LC/MS RT=2.15; [M+H].sup.+=305.1.
Step B: 6,7-Dichloroisoquinolin-3-amine and
5,6-dichloroisoquinolin-3-amine
##STR00917##
[0729] To sulfuric acid (4 mL, 75 mmol) was added
N-(3,4-dichlorobenzyl)-2,2-diethoxyacetimidamide (2 g, 6.6 mmol) at
room temperature. The reaction was heated to 40.degree. C. for 49
hours. TLC and LC/MS indicated the presence of product. The
reaction was cooled to room temperature and quenched with aq. NaOH
(.about.15 M) until the reaction mixture was .about.pH 7. The crude
product was extracted with ethyl acetate (2.times.50 mL) and the
organics were dried with MgSO.sub.4, filtered, and concentrated in
vacuo to yield the product. The crude product was purified by
chromatography (Biotage: 100% ethyl acetate to [90/10% ethyl
acetate/MeOH]) to yield approximately a 1:1 mixture of regioisomers
6,7-dichloroisoquinolin-1-amine and 5,6-dichloroisoquinolin-3-amine
(1.2 g, 5.64 mmol, 86.0% yield) as a dark yellow powder. The
regioisomers were carried on without separation. 1H NMR: 1H NMR
(500 MHz, DMSO-D.sub.6) .delta. ppm 8.90 (s, 1H), 8.83 (s, 1H),
8.12 (s, 1H), 7.89 (s, 1H), 7.84 (d, J=8.54 Hz, 1H), 7.28 (d,
J=8.85 Hz, 1H), 6.83 (s, 1H), 6.58 (s, 1H), 6.48 (s, 2H), 6.25 (s,
2H). MS (LC/MS) R.T.=1.59; [M+H].sup.+=213.0.
Step C: 6,7-Dichloro-3-isothiocyanatoisoquinoline and
5,6-dichloro-3-isothiocyanatoisoquinoline
##STR00918##
[0731] To 6,7-dichloroisoquinolin-3-amine and
5,6-dichloroisoquinolin-3-amine (0.410 g, 1.924 mmol) in
dichloromethane (20 mL) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (0.469 g, 2.021 mmol) and the
reaction mixture was stirred at 40.degree. C. for 4 hours. The
reaction was cooled to room temperature and chromatographed
(Biotage: 10-100% ethyl acetate/hexanes) to yield the separated
regioisomers, 6,7-dichloro-3-isothiocyanatoisoquinoline (0.2 g,
0.784 mmol, 40.7% yield) and
5,6-dichloro-3-isothiocyanatoisoquinoline (0.23 g, 0.902 mmol,
46.8% yield) as yellow solids.
5,6-dichloro-3-isothiocyanatoisoquinoline: 1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 9.10 (s, 1H), 7.87 (d, J=8.85 Hz, 1H), 7.82
(s, 1H), 7.66 (d, J=8.55 Hz, 1H). MS (LC/MS) R.T.=3.63;
[M+H].sup.+=255.0. 6,7-dichloro-3-isothiocyanatoisoquinoline: 1H
NMR (500 MHz, CDCl.sub.3) .delta. ppm 9.04 (s, 1H), 8.11 (s, 1H),
7.94 (s, 1H), 7.37 (s, 1H). MS (LC/MS) R.T.=3.42;
[M+H].sup.+=255.0.
Step C:
(R)--N-(6,7-Dichloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine
##STR00919##
[0733] To 6,7-dichloro-3-isothiocyanatoisoquinoline (0.13 g, 0.510
mmol) in DMF (10 mL) was added cesium carbonate (0.42 g, 1.27 mmol)
and (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.118 g,
0.515 mmol) at room temperature. The reaction was stirred at
70.degree. C. for 2 hours. The reaction was cooled to room
temperature and concentrated in vacuo. The crude urea was purified
by chromatography (Biotage: 85% CHCl.sub.3, 14% MeOH, 1%
NH.sub.4OH). The product was then treated with DMF (10 mL) and
N,N'-diisopropylcarbodiimide (0.238 mL, 1.529 mmol). The reaction
was heated to 90.degree. C. for 18 hours. The reaction was cooled
to room temperature and concentrated to yield the crude product,
which was purified by chromatography (Biotage: 85% CHCl.sub.3, 14%
MeOH, 1% NH.sub.4OH) to yield
(R)--N-(6,7-dichloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine (0.12 g, 0.312 mmol, 61.2% yield). 1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 9.03 (s, 1H), 8.87 (s, 1H), 7.96
(s, 1H), 7.83 (s, 1H), 7.24 (s, 1H), 3.88-4.06 (m, 1H), 3.60-3.74
(m, 1H), 3.42 (d, J=14.65 Hz, 1H), 2.82-3.15 (m, 5H), 2.23-2.34 (m,
1H), 2.18 (s, 1H), 1.72-1.87 (m, 1H), 1.48-1.70 (m, 2H). MS (LC/MS)
R.T.=1.63; [M+H].sup.+=377.1.
Example 281
(R)--N-(5,6-Dichloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine
##STR00920##
[0735] To 5,6-dichloro-3-isothiocyanatoisoquinoline (0.11 g, 0.431
mmol) in DMF (10 mL) was added cesium carbonate (0.351 g, 1.078
mmol) and (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.100 g, 0.435 mmol) at room temperature. The reaction was stirred
at 70.degree. C. for 2 hours. The reaction was cooled to room
temperature and concentrated in vacuo. The crude urea was purified
by chromatography (Biotage: 85% CHCl.sub.3, 14% MeOH, 1%
NH.sub.4OH). The product was then treated with DMF (10 mL) and
N,N'-diisopropylcarbodiimide (0.202 mL, 1.293 mmol). The reaction
was heated to 90.degree. C. for 18 hours. The reaction was cooled
to room temperature and concentrated to yield the crude product.
The crude product was purified by chromatography (Biotage: 85%
CHCl.sub.3, 14% MeOH, 1% NH.sub.4OH) to yield
(R)--N-(5,6-dichloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine (0.084 g, 0.218 mmol, 50.6% yield) as a
yellow powder. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 9.09
(s, 1H), 8.93 (s, 1H), 7.63-7.82 (m, 2H), 7.40 (d, J=8.55 Hz, 1H),
3.99 (d, J=9.16 Hz, 1H), 3.78 (d, J=8.85 Hz, 1H), 3.51 (d, J=14.65
Hz, 1H), 3.30 (d, J=14.65 Hz, 1H), 2.90-3.23 (m, 4H), 2.33-2.48 (m,
1H), 2.29 (s, 1H), 1.83-1.94 (m, 1H), 1.62-1.83 (m, J=42.12 Hz,
2H). MS (LC/MS) R.T.=1.57; [M+H].sup.+=377.1.
Example 282
(R)--N-(3,4-Dichloropyridin-1-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00921##
[0737]
(R)--N-(3,4-Dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine was prepared by the method of Example 267,
starting from 2-amino-3,4-dichloropyridine. 1H NMR (500 MHz,
DMSO-D.sub.6) .delta. ppm 9.10 (s, 1H), 8.08 (d, J=5.49 Hz, 1H),
7.13 (d, J=5.49 Hz, 1H), 3.86 (d, J=9.77 Hz, 1H), 3.60 (d, J=9.77
Hz, 1H), 2.96-3.05 (m, 2H), 2.77 (t, J=7.63 Hz, 2H), 2.66 (t,
J=7.78 Hz, 2H), 1.97-2.05 (m, 1H), 1.86-1.94 (m, 1H), 1.54-1.63 (m,
2H), 1.43-1.51 (m, 1H). MS (LC/MS) R.T.=0.78;
[M+H].sup.+=327.0.
Example 283
(R)--N-(3-Chloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oc-
tan]-2-amine
##STR00922##
[0739]
(R)--N-(3-Chloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine was prepared by the method of Example 267,
starting from 2-amino-3-chloropyridine. 1H NMR (500 MHz,
DMSO-D.sub.6) .delta. ppm 9.06 (s, 1 H), 8.14-8.19 (m, 1H),
7.74-7.79 (m, J=7.78, 1.83, 1.83, 1.68 Hz, 1H), 6.86-6.91 (m, 1H),
3.81-3.89 (m, 1H), 3.55-3.63 (m, 1H), 2.96-3.04 (m, 2H), 2.78 (t,
J=7.63 Hz, 2H), 2.67 (t, J=7.63 Hz, 2H), 1.96-2.02 (m, 1H),
1.86-1.92 (m, J=5.65, 3.20 Hz, 1H), 1.54-1.63 (m, J=6.87, 3.66,
3.51 Hz, 2H), 1.42-1.49 (m, 1 H). MS (LC/MS) R.T.=0.26;
[M+H].sup.+=293.0.
Example 284
(R)--N-(5-Chloro-3-fluoropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine
##STR00923##
[0741]
(R)--N-(5-Chloro-3-fluoropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine was prepared by the method of Example
267, starting from 2-amino-3-fluoro-5-chloropyridine. 1H NMR (500
MHz, DMSO-D.sub.6) .delta. ppm 8.81 (s, 1H), 8.05 (s, 1H), 7.79 (d,
J=10.07 Hz, 1H), 3.83 (d, J=9.46 Hz, 1H), 3.58 (d, J=9.46 Hz, 1H),
2.99 (s, 2H), 2.71-2.80 (m, 2H), 2.61-2.70 (m, 2H), 2.00 (s, 1H),
1.83-1.92 (m, 1H), 1.53-1.62 (m, 2H), 1.41-1.50 (m, 1H). MS (LC/MS)
R.T.=0.52; [M+].sup.+=311.0.
Example 285
(R)--N-(6-Chloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oc-
tan]-2-amine
##STR00924##
[0743]
(R)--N-(6-Chloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine was prepared by the method of Example 267,
starting from 2-amino-6-chloropyridine. 1H NMR (500 MHz,
DMSO-D.sub.6) .delta. ppm 8.33-8.42 (m, 1H), 7.60-7.68 (m, 1H),
6.94 (d, J=7.02 Hz, 1H), 6.72-6.81 (m, 1H), 3.86 (d, J=9.46 Hz,
1H), 3.57 (d, J=10.07 Hz, 1H), 2.97 (s, 2H), 2.69-2.78 (m, 2H),
2.63-2.68 (m, J=7.63, 7.63 Hz, 2H), 1.95-2.03 (m, 1H), 1.83-1.92
(m, 1H), 1.53-1.62 (m, 2H), 1.41-1.49 (m, 1H). MS (LC/MS)
R.T.=0.43; [M+H].sup.+=293.0.
Example 286
(R)--N-(4,6-Dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00925##
[0745]
(R)--N-(4,6-Dichloropyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine was prepared by the method of Example 267,
starting from 2-amino-4,6-dichloropyridine. 1H NMR (500 MHz,
DMSO-D6) .delta. ppm 8.43 (s, 1H), 7.13 (s, 1H), 6.84 (s, 1H), 3.86
(d, J=9.77 Hz, 1H), 3.59 (d, J=10.07 Hz, 1H), 2.98 (s, 2H),
2.58-2.86 (m, 4H), 1.94-2.13 (m, 1H), 1.78-1.95 (m, 1H), 1.36-1.65
(m, 3H). MS (LC/MS) R.T.=0.87; [M+H].sup.+=327.0.
Example 287
(R)--N-(2-Methoxy-3-4'-bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine
##STR00926##
[0746] Step A: N-(2,4-Dichlorobenzyl)-2,2-diethoxyacetimidamide
##STR00927##
[0748] To 4-bromopyridin-2-amine (0.5 g, 2.8 mmol),
2-methoxypyridin-3-ylboronic acid (0.52 g, 3.4 mmol) in DMF (25 mL)
was added 1N sodium carbonate (10 mL, 2.3 mmol), followed by
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex (0.21 g, 0.26 mmol). The reaction mixture
was stirred for 3 hours at 85.degree. C. and then cooled to room
temperature. The product was extracted with ethyl acetate
(2.times.50 mL), dried with MgSO.sub.4, filtered, and concentrated
in vacuo. The crude product was purified by chromotography
(Biotage: 100 to 90/10% ethyl acetate-ethyl acetate/methanol) to
yield 2-methoxy-3,4'-bipyridin-2'-amine (0.53 g, 2.63 mmol, 93%
yield) as a brown powder. The product was taken directly to the
next step.
Step B: 2'-Isothiocyanato-2-methoxy-3,4'-bipyridine
##STR00928##
[0750] To 2-methoxy-3,4'-bipyridin-2'-amine (0.53 g, 2.63 mmol) in
dichloromethane (20 mL) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (0.62 g, 2.7 mmol) and the
reaction mixture was stirred at 40.degree. C. for 4 hours. The
reaction was cooled to room temperature and chromatographed
(Biotage: 10-100% ethyl acetate/hexanes) to yield
2'-isothiocyanato-2-methoxy-3,4'-bipyridine (0.46 g, 1.9 mmol,
71.8% yield). 1H NMR (500 MHz, DMSO-D6) .delta. ppm 8.67 (d, J=2.44
Hz, 1 H), 8.26 (dd, J=4.88, 1.53 Hz, 1H), 8.15 (dd, J=8.24, 2.44
Hz, 1H), 7.89 (dd, J=7.32, 1.53 Hz, 1H), 7.48 (d, J=8.24 Hz, 1H),
7.15 (dd, J=7.32, 4.88 Hz, 1H), 3.91 (s, 3H). MS (LC/MS) R.T.=2.87;
[M+H].sup.+=244.9.
Step D:
(R)--N-(2-Methoxy-3-4'-bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine
##STR00929##
[0752] To 2'-isothiocyanato-2-methoxy-3,4'-bipyridine (0.09 g, 0.37
mmol) in DMF (20 mL) was added Et.sub.3N (0.11 mL, 0.81 mmol) and
(S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.09 g, 0.37
mmol) at room temperature. The reaction was stirred at 70.degree.
C. for 2 hours. The reaction was cooled to room temperature and
concentrated in vacuo. The crude urea was purified by
chromatography (biotage: 85% CHCl.sub.3, 14% MeOH, 1% NH.sub.4OH)
to yield the pure urea intermediate. The product was then treated
with DMF (20 mL) and N,N'-diisopropylcarbodiimide (0.17 mL, 1.1
mmol). The reaction was heated to 90.degree. C. for 18 hours. The
reaction was cooled to room temperature and concentrated to yield
the crude product. The crude product was purified by chromatography
(Biotage: 85% CHCl.sub.3, 14% MeOH, 1% NH.sub.4OH) and the
product-containing fractions were combined. LC/MS and 1HNMR
indicated some impurities may be present. The impure product was
subjected to reverse phase HPLC to yield
(R)--N-(2-methoxy-3,4'-bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine (0.02 g, 0.05 mmol, 14.05% yield) as a
white powder. 1H NMR (500 MHz, DMSO-D6) .delta. ppm 9.07 (s, 1H),
8.43 (s, 1H), 8.17 (dd, J=4.73, 1.68 Hz, 1H), 7.79 (d, J=7.32 Hz,
2H), 7.10 (dd, J=7.32, 4.88 Hz, 1H), 6.79-6.92 (m, 1H), 3.76-3.97
(m, 4H), 3.51-3.66 (m, 1H), 2.92-3.09 (m, 2H), 2.59-2.82 (m, 4H),
1.85-2.03 (m, 2H), 1.53-1.71 (m, 2H), 1.35-1.49 (m, 1H). MS (LC/MS)
R.T.=1.05; [M+H].sup.+=366.1.
Example 288
(R)--N-(Benzo[d]oxazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oct-
an]-2-amine
##STR00930##
[0753] Step A: Benzo[d]oxazol-2-amine
##STR00931##
[0755] An oven dried round bottom flask was charged with
di(1H-imidazol-1-yl)methanimine (500 mg, 3.10 mmol), 2-aminophenol
(188 mg, 1.724 mmol) and anhydrous THF (20 ml) at room temperature.
The resulting suspension was refluxed under N.sub.2 for 2 hr to
give complete conversion based on LC/MS. The solvent was removed in
vacuo and the residue was purified on a Biotage Flash Collector,
eluting with 30-80% EtOAc/Hexane (1200 ml) to afford the expected
product, benzo[d]oxazol-2-amine (200 mg, 1.5 mmol, 87% yield), as a
white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 6.20
(br. s., 2H) 7.02-7.11 (m, 1H) 7.17-7.22 (m, 1H) 7.29 (d, J=7.53
Hz, 1H) 7.36 (d, J=7.03 Hz, 1H). MS (LC/MS) R.T.=1.05;
[M+H].sup.+=134.96.
Step B: Dimethyl benzo[d]oxazol-2-ylcarbonimidodithioate
##STR00932##
[0757] To a colorless solution of benzo[d]oxazol-2-amine (200 mg,
1.491 mmol) in DMF (10 ml) was added sodium hydroxide (20N, 149
.mu.L, 2.98 mmol), to give a green suspension. The mixture was
stirred for 15 min at room temperature. Carbon disulfide (225
.mu.L, 3.73 mmol) was added to give a dark brown solution. The
reaction was stirred for 15 min at room temperature, followed by
the addition of sodium hydroxide (20N, 149 .mu.L, 2.98 mmol) and
stirred for an additional 10 min. Iodomethane (224 .mu.L, 3.58
mmol) was then added dropwise. A green solid precipitated after 12
min. The reaction stirred for a further 2 hr. The solid was
collected by filtration, washed with DMF (2.times.1 ml), H.sub.2O
(2.times.1 ml), dried under the house vacuum for 30 min and further
dried in an oven in vacuo over night to afford the expected
product, dimethyl benzo[d]oxazol-2-ylcarbonimidodithioate (258.5
mg, 1.085 mmol, 72.7% yield), as a white solid. 1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 2.70 (s, 6H) 7.24-7.34 (m, 2H) 7.45-7.50
(m, 1H) 7.66-7.74 (m, 1H). MS (LC/MS) R.T.=1.76,
[M+H].sup.+=238.96.
Step C:
(R)--N-(Benzo[d]oxazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine
[0758] A 10 ml-vial was charged with
(S)-3-(aminomethyl)quinuclidin-3-ol.2HCl salt (69.5 mg, 0.361
mmol), DMF (2 ml), DIEA (0.063 mL, 0.361 mmol) and Cs.sub.2CO.sub.3
(235 mg, 0.722 mmol) at room temperature, followed by dimethyl
benzo[d]oxazol-2-ylcarbonimidodithioate (86 mg, 0.361 mmol). The
resulting suspension was stirred at room temperature for 1 hr.
LC/MS then indicated consumption of starting material. The mixture
was diluted with MeOH and purified by preparative HPLC to afford
the expected product,
(R)--N-(benzo[d]oxazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oc-
tan]-2-amine (101.5 mg, 0.323 mmol, 90% yield), as a tan gum. 1H
NMR (400 MHz, Acetone-d.sub.6) .delta. ppm 2.07-2.14 (m, 2H) 2.20
(ddd, J=8.78, 5.27, 3.26 Hz, 2H) 2.33-2.45 (m, 1H) 2.62 (d, J=2.26
Hz, 1H) 3.34-3.47 (m, 3H) 3.48-3.58 (m, 1H) 3.75-3.88 (m, 2H) 4.15
(d, J=10.54 Hz, 1H) 4.32 (d, J=10.54 Hz, 1 H) 7.13-7.26 (m, 2H)
7.41 (td, J=3.70, 1.63 Hz, 1H) 9.24 (br, s, 1H). MS (LC/MS)
R.T.=0.792, [M+H].sup.+=299.17.
Example 289
(R)--N-(5-Chlorobenzo[d]oxazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine
##STR00933##
[0759] Step A: Dimethyl
5-chlorobenzo[d]oxazol-2-ylcarbonimidodithioate
##STR00934##
[0761] To a brown solution of 5-chlorobenzo[d]oxazol-2-amine (700
mg, 4.15 mmol) in DMF (5 ml) was added sodium hydroxide (20N, 415
.mu.L, 8.30 mmol) to give a grey suspension. The mixture was
stirred for 15 min at room temperature. Carbon disulfide (626
.mu.L, 10.38 mmol) was added at room temperature to give a brown
solution. The mixture was stirred for 15 min at room temperature,
then sodium hydroxide (20N, 208 .mu.L, 4.16 mmol) was added. After
10 min, iodomethane (623 .mu.L, 9.97 mmol) was added dropwise. A
grey solid came out from solution. The reaction was further stirred
at room temperature for 2 hr. The solid was collected by
filtration, washed with DMF/H.sub.2O (50:50, 2.times.2 ml), dried
under house vacuum for 30 min and further dried in an oven at
65.degree. C. under vacuum for 11/2 hr to afford the expected
product, dimethyl 5-chlorobenzo[d]oxazol-2-ylcarbonimidodithioate
(780 mg, 2.86 mmol, 68.9% yield), as an off-white solid which was
pure enough to be used in next step. 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 2.69 (s, 6H) 7.37 (dd, J=8.53, 2.26 Hz, 1H) 7.66 (d,
J=9.03 Hz, 1H) 7.75 (d, J=1.76 Hz, 1H). MS (LC/MS) R.T.=1.44;
[M+H].sup.+=272.9.
Step B:
(R)--N-(5-Chlorobenzo[d]oxazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine
##STR00935##
[0763] A 10 ml vial was charged with
(S)-3-(aminomethyl)quinuclidin-3-ol.2HCl salt (106 mg, 0.550 mmol),
DMF (2 ml), DIEA (0.096 mL, 0.550 mmol), and Cs.sub.2CO.sub.3 (358
mg, 1.100 mmol) at room temperature, followed by dimethyl
5-chlorobenzo[d]oxazol-2-ylcarbonimidodithioate (150 mg, 0.550
mmol). The resulting suspension was stirred at room temperature for
1 hr. LC/MS then indicated consumption of starting material. The
reaction mixture was diluted with MeOH and purified by preparative
HPLC to afford the expected product,
(R)--N-(5-chlorobenzo[d]oxazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octan]-2-amine (106.5 mg, 0.3 mmol, 53.0% yield), as
a white solid. 1H NMR (400 MHz, Acetone-d.sub.6) .delta. ppm
2.08-2.30 (m, 3H) 2.35-2.48 (m, 1H) 2.60-2.73 (m, 1H) 3.48 (qd,
J=7.53, 7.28 Hz, 3H) 3.54-3.68 (m, 1H) 3.79-4.00 (m, 2H) 4.18 (d,
J=10.54 Hz, 1H) 4.35 (d, J=10.54 Hz, 1H) 7.19 (dd, J=8.53, 2.01 Hz,
1H) 7.30-7.47 (m, 2 H) 9.10 (br. s., 1H). MS (LC/MS) R.T.=1.56;
[M+H].sup.+=333.13.
Example 290
(R)--N-(Oxazolo[4,5-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.-
2.2]octan]-2-amine
##STR00936##
[0764] Step A: Oxazolo[4,5-b]pyridin-2-amine
##STR00937##
[0766] An oven dried round bottom flask was charged with
di(1H-imidazol-1-yl)methanimine (500 mg, 3.10 mmol),
2-aminopyridin-3-ol (171 mg, 1.551 mmol) and anhydrous THF (20 ml)
at room temperature. The resulting suspension was refluxed under
N.sub.2 for 1 hr. LC/MS indicated complete consumption of starting
material. The solvent was removed in vacuo and the residue was used
in the next step without further purification. MS (LC/MS)
R.T.=0.235; [M+H].sup.+=136.09.
Step B: Dimethyl oxazolo[4,5-b]pyridin-2-ylcarbonimidodithioate
##STR00938##
[0768] To the crude oxazolo[4,5-b]pyridin-2-amine (811 mg, 6 mmol)
from step A, in DMF (12 ml), was added NaOH (20 N, 600 .mu.L, 12.00
mmol) to give a tan solution which was stirred for 15 min at room
temperature. Carbon disulfide (904 .mu.L, 15.00 mmol) was then
added to give an orange solution. The mixture was stirred for 15
min at room temperature, then NaOH (20 N, 600 .mu.L, 12.00 mmol)
was added and the stirring continued for 10 min to give a dark red
solution. Iodomethane (900 .mu.L, 14.40 mmol) was added dropwise,
resulting in a yellow solid precipitating after 1 hr to give
.about.80% conversion. The mixture was diluted with MeOH and
purified via preparative HPLC to afford the expected product,
dimethyl oxazolo[4,5-b]pyridin-2-ylcarbonimidodithioate (35 mg,
0.146 mmol, 2.4% yield), as a white solid. 1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 2.71 (s, 6H) 7.22 (dd, J=8.03, 5.02 Hz, 1H)
7.72 (dd, J=8.03, 1.25 Hz, 1H) 8.49 (dd, J=5.02, 1.51 Hz, 1H). MS
(LC/MS) R.T.=1.358; [M+H]=240.04.
Step C:
(R)--N-(Oxazolo[4,5-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine
##STR00939##
[0770] A 10 ml vial was charged with
(S)-3-(aminomethyl)quinuclidin-3-ol.2HCl salt (8.53 mg, 0.044
mmol), DMF (2 ml), DIEA (7.74 .mu.L, 0.04 mmol) and
Cs.sub.2CO.sub.3 (28.9 mg, 0.089 mmol) at room temperature,
followed by dimethyl oxazolo[4,5-b]pyridin-2-ylcarbonimidodithioate
(10.6 mg, 0.044 mmol). The resulting suspension was stirred at room
temperature for 1 hr. LC/MS indicated complete consumption of
starting material. The reaction mixture was diluted with MeOH and
purified via preparative HPLC to afford the expected product,
(R)--N-(oxazolo[4,5-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine (13 mg, 0.038 mmol, 86% yield), as a white
solid. 1H NMR (400 MHz, Acetone-d.sub.6) .delta. ppm 1.54-1.60 (m,
1H) 1.71-1.77 (m, 2H) 2.16-2.24 (m, 1H) 2.77-2.82 (m, 2H) 2.89 (t,
J=7.91 Hz, 4H) 3.13-3.25 (m, 2H) 3.90 (d, J=10.29 Hz, 1 H) 4.22 (d,
J=10.29 Hz, 1H) 7.13 (dd, J=8.03, 5.02 Hz, 1H) 7.70 (dd, J=7.91,
1.13 Hz, 1H) 8.27 (dd, J=5.14, 1.13 Hz, 1H) 9.11 (br. s., 1H). MS
(LC/MS) R.T.=0.443; [M+H].sup.+=300.16.
Example 291
(2R)--N-(6,8-Dimethyl-3-isoquinolinyl)-4'H-spiro[4-azabicyclo[2.2.2]octane-
-2,5'-[1,3]oxazol]-2'-amine
##STR00940##
[0771] Step A: 6-Methylisoquinolin-3-amine
##STR00941##
[0773] To a solution of methyl 2,2-diethoxyacetimidate (1.1 g, 6.82
mmol) in methanol (8 mL) was added p-tolylmethanamine (0.788 g,
6.50 mmol) dropwise at ambient temperature. The reaction flask was
then placed into a preheated oil-bath and stirred at 70.degree. C.
for 16 h, then removed and allowed cooled. The volatiles were
removed under reduced pressure and the crude material was added
dropwise to sulfuric acid (5 mL) at ambient temperature. The
reaction mixture was stirred for 72 h, then the flask was placed
into an ice-water bath, diluted with water (50 mL), and slowly
neutralized to pH=10 with sodium hydroxide (10 N). As the reaction
mixture became basic, a grey precipitate formed. This precipitate
was filtered, washed with water, and dried to afford
6-methylisoquinolin-3-amine (0.65 g, 63%), as a grey powder. 1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 8.73 (s, 1H), 7.69 (d, J=8.28
Hz, 1H), 7.29 (s, 1H), 7.00 (d, J=8.28 Hz, 1H), 5.81 (s, 1H), 2.40
(s, 3H). MS (LC/MS) R.T.=1.37; [M+H].sup.+=159.10.
Step B: 3-Isothiocyanato-6-methylisoquinoline
##STR00942##
[0775] To a solution of 1,1'-thiocarbonyldipyridin-2(1H)-one (220
mg, 0.948 mmol) in dichloromethane (10 mL) at ambient temperature
was added 6-methylisoquinolin-3-amine (125 mg, 0.790 mmol). The
reaction mixture was placed into a preheated oil-bath and stirred
at 40.degree. C. for 18 h, then removed from the oil-bath and
cooled to ambient temperature. The mixture was concentrated and the
crude material was purified by silica gel chromatography (5-30%
ethyl acetate in hexanes) to afford
3-isothiocyanato-6-methylisoquinoline (75 mg, 0.375 mmol, 47.4%
yield), as an off-white solid. 1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 9.03 (s, 1H), 7.88 (d, J=8.24 Hz, 1 H), 7.56 (s, 1H), 7.46 (dd,
J=8.24, 1.53 Hz, 1H), 7.39 (s, 1H) 2.57 (s, 3H). MS (LC/MS)
R.T.=1.92; [M+H].sup.+=201.13.
Step C:
(R)--N-(6-Methylisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00943##
[0777] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (65.8
mg, 0.285 mmol) in N,N-dimethylformamide (6 mL) was added
triethylamine (0.090 mL, 0.63 mmol) and
3-isothiocyanato-6-methylisoquinoline (57 mg, 0.285 mmol). The
suspension was placed into a preheated oil-bath and stirred at
70.degree. C. for 2 h and 30 min. N,N'-diisopropylcarbodiimide
(0.177 mL, 1.14 mmol) was then added and the mixture was stirred at
85.degree. C. for 16 h. The mixture was concentrated and purified
by silica gel chromatography (0-40% [9:1 methanol:ammonium
hydroxide] in chloroform) followed by purification by reverse phase
preparatory HPLC (0-40% TFA-methanol-water). The solution of
product was filtered through UCT Clean-up CHQAX15M25 cartridge with
MeOH (3.times.10 ml) and concentrated to afford the expected
product,
(S)-1-((3-hydroxyquinuclidin-3-yl)methyl)-3-(6-methylisoquinolin-3-yl)thi-
ourea, as a tan gum. 1H NMR (400 MHz, MeOD) .delta. ppm 9.11 (s,
1H), 8.02 (d, J=8.53 Hz, 1H), 7.80 (s, 1H), 7.74 (s, 1H), 7.47 (d,
J=8.53 Hz, 1H), 4.47 (d, J=10.54 Hz, 1H), 4.31 (d, J=10.54 Hz, 1H),
4.10 (d, J=14.81 Hz, 1H), 3.92 (d, J=14.81 Hz, 1H), 3.53-3.71 (m,
2H), 3.27-3.51 (m, 3H), 2.84 (br.s, 1H), 2.55 (s, 3H), 2.48 (m,
1H),) 2.10-2.30 (m, 3H). MS (LC/MS) R.T.=1.24;
[M+H].sup.+=323.2.
Example 292
(R)--N-(6-Bromoisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00944##
[0778] Step A: N-(4-Bromobenzyl)-2,2-diethoxyacetimidamide
##STR00945##
[0780] (4-Bromophenyl)methanamine hydrochloride (2.359 g, 10.39
mmol) and sodium methoxide (2.376 mL, 10.39 mmol) were added to a
solution of methyl 2,2-diethoxyacetimidate (3.35 g, 20.78 mmol) in
methanol (10 ml). The cloudy mixture was heated at 70 C for 1.5 h
and the resulting yellow mixture was concentrated. The residue was
purified by silica gel with 100% ethyl acetate to give a yellow
viscous oil (2.04 g, 62%). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 7.47 (2H, d, J=8.56 Hz), 7.23 (2 H, d, J=8.06 Hz), 6.76
(1H, br. s.), 5.31 (1H, br. s.), 4.94 (1H, br. s.), 4.45 (2H, br.
s.), 3.47-3.77 (4H, m), 1.26 (6H, t, J=7.05 Hz). LCMS:R.T.=2.12;
[M+2].sup.+=317.2.
Step B: 6-Bromoisoquinolin-3-amine
##STR00946##
[0782] N-(4-bromobenzyl)-2,2-diethoxyacetimidamide (1.53 g, 4.85
mmol) in sulfuric acid (4 mL, 95-98%) was heated at 40 C for 14 h.
The mixture was neutralized with 1 M NaOH to pH 7 and the resulting
suspension was filtered. The residue was purified by silica gel
chromatography with 20-55% ethyl acetate in hexanes. The desired
fractions were concentrated to give a brownish yellow solid (0.434
g, 40%). LCMS:R.T.=1.62; [M+2].sup.+=225.1.
Step C: 6-Bromo-3-isothiocyanatoisoquinoline
##STR00947##
[0784] To a solution of 1,1'-thiocarbonyldipyridin-2(1H)-one (0.251
g, 1.080 mmol) in dichloromethane at room temperature was added
6-bromoisoquinolin-3-amine (0.241 g, 1.080 mmol). The solution was
stirred at room temperature for 3 hours.
[0785] LC/MS indicated formation of the desired product. The deep
orange solution was purified by silica gel chromatography (0-10%
ethyl acetate-hexanes) to afford:
6-bromo-3-isothiocyanatoisoquinoline (0.1 g, 0.377 mmol, 35% yield)
as a yellow oil. R.T.=2.54; [M+H].sup.+=267.04.
Step D:
(R)--N-(6-bromoisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00948##
[0787] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.086 g, 0.377 mmol) in N,N-dimethylformamide (15 mL) was added
Cs.sub.2CO.sub.3 (0.307 g, 0.943 mmol) and
6-bromo-3-isothiocyanatoisoquinoline (0.1 g, 0.377 mmol). The
suspension was stirred at room temperature for 30 minutes.
N,N'-Diisopropylcarbodiimide (0.176 mL, 1.132 mmol) was then added
and the mixture was stirred for a further 18 hours. The mixture was
concentrated and purified by silica gel chromatography (5-25%
[9.5:0.5 methanol:ammonium hydroxide]-ethyl acetate) to afford
(R)--N-(6-bromoisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (0.054 g, 0.135 mmol, 36% yield) as a yellow
solid. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.04 (1H, s),
7.80-8.05 (2H, m), 7.55 (1H, dd, J=8.81, 1.76 Hz), 7.37 (1H, br.
s.), 4.10 (1H, d, J=10.58 Hz), 3.87 (1H, d, J=10.83 Hz), 3.68-3.77
(1H, m), 3.56-3.67 (1H, m), 3.29-3.49 (4H, m), 2.45 (1H, br. s.),
2.28-2.41 (1H, m), 1.86-2.15 (3H, m). LCMS:R.T.=1.76;
[M+].sup.+=387.21.
Example 293
(R)--N-(7-Bromoisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00949##
[0788] Step A: N-(3-Bromobenzyl)-2,2-diethoxyacetimidamide
##STR00950##
[0790] (3-Bromophenyl)methanamine hydrochloride (3.62 g, 15.64
mmol) and sodium methoxide (3.58 mL, 15.64 mmol) were added to a
solution of methyl 2,2-diethoxyacetimidate (5.042 g, 31.3 mmol) in
methanol (15 mL). The cloudy mixture was heated at 70 C for 1.5 h
and the resulting yellow mixture was concentrated. The residue was
purified by silica gel chromatography with 100% ethyl acetate. The
desired fractions were concentrated to give a yellow viscous oil
(2.5 g, 51%).
[0791] LCMS:R.T.=2.11; [M+2].sup.+=317.06.
Step B: 7-Bromoisoquinolin-3-amine and
5-bromoisoquinolin-3-amine
##STR00951##
[0793] N-(3-Bromobenzyl)-2,2-diethoxyacetimidamide (2.5 g, 7.93
mmol) in sulfuric acid (5 mL, 95-98%) was heated at 40 C for 54 h.
The mixture was neutralized with 10 M NaOH aqueous to pH 7 and the
resulting suspension was filtered. The residue was purified by
silica gel chromatography with 20-55% ethyl acetate in hexanes,
then 100% ethyl acetate. The fractions were concentrated to give a
brown solid containing a mixture of products. (1.0 g, 57%).
LCMS:R.T.=1.56; [M+2].sup.+=225.1. The mixture was used as is for
the next step.
Step C: 7-Bromo-3-isothiocyanatoisoquinoline and
5-bromo-3-Isothiocyanatoisoquinoline
##STR00952##
[0795] To a solution of 1,1'-thiocarbonyldipyridin-2(1H)-one 1.145
g, 4.93 mmol) in dichloromethane at room temperature was added the
mixture of 7-bromoisoquinolin-3-amine and
5-bromoisoquinolin-3-amine from step B (1.0 g, 4.5 mmol). The
orange solution was stirred at room temperature for 18 hours. LCMS
indicated the formation of product. The deep orange solution was
purified by silica gel chromatography (0-5% ethyl acetate-hexanes).
The first product fractions were combined and concentrated in vacuo
to afford 7-bromo-3-isothiocyanatoisoquinoline (0.27 g, 0.377 mmol,
22% yield) as a yellow solid. .sup.1H NMR (400 MHz, Acetone)
.delta. ppm 9.19 (1H, s), 8.41 (1H, s), 7.86-8.02 (2H, m), 7.76
(1H, s). R.T.=4.28; [M+H].sup.+=267.04. The second product
fractions were combined and concentrated in vacuo to afford
5-bromo-3-isothiocyanatoisoquinoline (0.25 g, 0.377 mmol, 21%
yield) as a yellow solid. .sup.1H NMR (400 MHz, Acetone) .delta.
ppm 9.24 (1H, s), 8.23 (1H, d), 8.15 (1H, d), 7.81 (1H, s), 7.64
(1H, t). LCMS:R.T.=4.61; [M+H].sup.+=267.04.
Step D:
(R)--N-(7-Bromoisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine
##STR00953##
[0797] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.207 g, 0.905 mmol) in N,N-dimethylformamide (20 mL) was added
Cs.sub.2CO.sub.3 (0.737 g, 2.263 mmol) and
7-bromo-3-isothiocyanatoisoquinoline (0.24 g, 0.905 mmol). The
suspension was stirred at room temperature for 30 minutes.
N,N'-Diisopropylcarbodiimide (0.423 mL, 2.72 mmol) was then added
and the mixture was stirred at room temperature for 18 hours. The
mixture was concentrated and purified by silica gel chromatography
using 5-15% [9:1 methanol:ammonium hydroxide] in ethyl acetate. The
desired fractions were concentrated and further purified using
5-15% [9.5:0.5 methanol:ammonium hydroxide] in ethyl acetate to
afford
((R)--N-(7-bromoisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2-
.2]octan]-2-amine (0.061 g, 0.156 mmol, 17% yield) as a yellow
solid. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.96 (1H, s), 8.12
(1H, s), 7.66 (2H, s), 7.32 (1H, s), 4.01 (1H, d), 3.73 (1H, d),
3.35-3.42 (1H, m), 3.22-3.29 (1H, m), 2.84-3.17 (4H, m), 2.13-2.35
(2H, m), 1.62-1.96 (3H, m). LCMS:R.T.=1.76; [M+].sup.+=387.21.
Example 294
((R)--N-(5-Bromoisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00954##
[0799] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride
(0.207 g, 0.905 mmol) in N,N-dimethylformamide (20 mL) was added
Cs.sub.2CO.sub.3 (0.737 g, 2.263 mmol) and
5-bromo-3-isothiocyanatoisoquinoline (0.24 g, 0.905 mmol). The
suspension was stirred at room temperature for 30 minutes.
N,N'-Diisopropylcarbodiimide (0.423 mL, 2.72 mmol) was then added
and the mixture was stirred at room temperature for 18 hours. The
mixture was concentrated and purified by silica gel chromatography
(5-15% [9.5:0.5 methanol:ammonium hydroxide]-ethyl acetate). The
desired fractions were concentrated and further purified using
5-15% [9.5:0.5 methanol:ammonium hydroxide] in ethyl acetate to
afford
(R)--N-(5-bromoisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (0.248 g, 0.634 mmol, 70% yield) as a yellow
solid. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.99 (1H, s), 7.91
(2H, dd), 7.55 (1H, br. s.), 7.28 (1H, t), 3.96 (1H, d), 3.65 (1H,
d), 3.17-3.26 (1H, m), 3.03-3.13 (1H, m), 2.70-2.99 (4H, m),
2.03-2.30 (2H, m), 1.47-1.87 (3H, m). LCMS:R.T.=1.69;
[M+2].sup.+=389.21.
Example 295
(2R)--N-(6,8-Dimethyl-3-isoquinolinyl)-4'H-spiro[4-azabicyclo[2.2.2]octane-
-2,5'-[1,3]oxazol]-2'-amine
##STR00955##
[0800] Step A: 6,8-Dimethylisoquinolin-3-amine
##STR00956##
[0802] To a solution of methyl 2,2-diethoxyacetimidate (1.5 g, 9.3
mmol) in methanol (4.9 mL) was added
(2,4-dimethylphenyl)methanamine (1.2 g, 8.9 mmol) dropwise at
ambient temperature. The reaction flask was then placed into a
preheated oil-bath and stirred at 70.degree. C. for 16 h, then
cooled and the volatiles removed under reduced pressure. The crude
material was added dropwise to sulfuric acid (19.7 mL) at ambient
temperature and stirred for 72 h. The flask was then placed into an
ice-water bath, diluted with water (50 mL), and slowly neutralized
to pH=10 with sodium hydroxide (10 N). As the reaction mixture
became basic, an orange precipitate formed. This precipitate was
filtered, washed with water, and dried to afford
6,8-dimethylisoquinolin-3-amine (1.37 g, 7.95 mmol, 90%). 1H NMR
(400 MHz, MeOD) .delta. ppm 8.82 (s, 1H), 7.17 (s, 1H), 6.90 (s,
1H), 6.72 (s, 1H), 2.61 (s, 3H), 2.37 (s, 3H). MS (LC/MS)
R.T.=0.77; [M+H].sup.+=173.15.
Step B: 3-Isothiocyanato-6,8-dimethylisoquinoline
##STR00957##
[0804] To a solution of 1,1'-thiocarbonyldipyridin-2(1H)-one (1.35
g, 5.8 mmol) in dichloromethane (19 mL) at ambient temperature was
added 6,8-dimethylisoquinolin-3-amine (1 g, 5.8 mmol). The reaction
mixture was placed into a preheated oil-bath and stirred at
40.degree. C. for 18 h, then cooled, concentrated, and the crude
material was purified by silica gel chromatography (10-35% ethyl
acetate in hexanes) to afford
3-isothiocyanato-6,8-dimethylisoquinoline (93.7 mg, 0.437 mmol, 8%)
as a yellow solid. 1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 9.20
(s, 1H), 7.36-7.44 (m, 2H), 7.23 (s, 1H), 2.75 (s, 3H), 2.51 (s,
3H). MS (LC/MS) R.T.=2.03; [M+H].sup.+=215.1.
Step C:
(2R)--N-(6,8-Dimethyl-3-isoquinolinyl)-4'H-spiro[4-azabicyclo[2.2.-
2]octane-2,5'-[1,3]oxazol]-2'-amine
##STR00958##
[0806] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (99
mg, 0.43 mmol) in N,N-dimethylformamide (1.4 mL) was added
triethylamine (0.18 mL, 1.3 mmol) and
3-isothiocyanato-6,8-dimethylisoquinoline (93 mg, 0.43 mmol). The
suspension was placed into a preheated oil-bath and stirred at
70.degree. C. for 2 h and 30 min. N,N'-Diisopropylcarbodiimide
(0.27 mL, 1.7 mmol) was then added and the mixture was stirred at
85.degree. C. for 16 h. The mixture was concentrated and purified
by silica gel chromatography (0-40% [9:1 methanol:ammonium
hydroxide] in chloroform) followed by purification by reverse phase
preparative HPLC (0-40% [0.1% TFA]-methanol-water) to afford
(2R)--N-(6,8-dimethyl-3-isoquinolinyl)-4'H-spiro[4-azabicyclo[2.2.2]octan-
e-2,5'-[1,3]oxazol]-2'-amine as the trifluoroacetic acid salt (24
mg, 0.053 mmol, 12% yield) as a white solid. 1H NMR (400 MHz, MeOD)
.delta. ppm 9.29 (s, 1H), 7.54 (s, 1H), 7.43 (s, 1H), 7.34 (s, 1H),
4.38 (d, J=11.04 Hz, 1H), 4.17 (d, J=11.04 Hz, 1H), 3.94-4.10 (m,
1H), 3.86 (dd, J=15.06, 2.26 Hz, 1H), 3.51-3.67 (m, 1H), 3.37-3.51
(m, 3H), 2.77 (s, 3H), 2.73 (d, J=3.51 Hz, 1H), 2.48-2.57 (m, 3H),
2.30-2.46 (m, 1H), 1.94-2.28 (m, 3H). MS (LC/MS) R.T.=0.90;
[M+H].sup.+=337.38.
Example 296
(R)--N-(3,4'-Bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oc-
tan]-2-amine
##STR00959##
[0808]
(R)--N-(3,4'-Bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine was prepared from 4-bromopyridin-2-amine,
pyridin-3-ylboronic acid, and
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex by following the general procedures of
Example 28, Steps A-B. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 8.95 (br. s., 2H), 8.66 (d, J=4.6 Hz, 1H), 8.31 (d, J=5.5 Hz,
1H), 8.14 (br. s., 1H), 7.54 (d, J=7.6 Hz, 1H), 7.26 (br. s., 2H),
3.85 (br. s., 1H), 3.58 (s, 1H), 3.00 (br. s., 2H), 2.67 (br. s.,
4H), 1.90 (br. s., 2H), 1.59 (br. s., 2H), 1.45 (br. s., 1H). MS
(LC/MS) R.T.=0.12; [M+H].sup.+=336.18.
Example 297
(R)--N-(5-Chloro-3,4'-bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine
##STR00960##
[0810]
(R)--N-(5-Chloro-3,4'-bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine was prepared from
4-bromopyridin-2-amine, 5-chloropyridin-3-ylboronic acid, and
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex by following the general procedures of
Example 28, Steps A-B. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 8.92 (s, 2H), 8.71 (d, J=2.1 Hz, 1H), 8.19-8.39 (m, 2H), 7.30
(d, J=4.3 Hz, 2H), 3.77-3.95 (m, 1H), 3.59 (d, J=10.1 Hz, 1H), 3.01
(d, J=4.9 Hz, 2H), 2.61-2.86 (m, 4H), 2.00 (s, 1H), 1.83-1.96 (m,
1H), 1.55-1.68 (m, 2H), 1.40-1.54 (m, 1H). MS (LC/MS) R.T.=0.91;
[M+H].sup.+=370.09.
Example 298
(R)--N-(6-Methoxy-3,4'-bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine
##STR00961##
[0812]
(R)--N-(6-Methoxy-3,4'-bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octan]-2-amine was prepared from
4-bromopyridin-2-amine, 6-methoxypyridin-3-ylboronic acid, and
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex by following the general procedures of
Example 28, Steps A-B. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 8.79-9.21 (m, 1H), 8.46-8.66 (m, 1H), 8.26 (s, 1H), 7.97-8.14
(m, 1H), 6.99-7.27 (m, 2H), 6.83-6.99 (m, 1H), 3.92 (s, 4H),
3.49-3.62 (m, 1H), 2.99 (d, J=4.3 Hz, 2H), 2.66 (d, J=7.6 Hz, 4H),
1.79-2.05 (m, 2H), 1.33-1.63 (m, 3H). MS (LC/MS) R.T.=0.98;
[M+H].sup.+=366.17.
Example 299
(R)--N-(6-Fluoro-3,4'-bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine
##STR00962##
[0814]
(R)--N-(6-Fluoro-3,4'-bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine was prepared from
4-bromopyridin-2-amine,
2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine,
and 1,1'-bis(diphenylphosphino) ferrocene-palladium(II)dichloride
dichloromethane complex by following the general procedures of
Example 28, Steps A-B. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 8.90-9.17 (m, 1H), 8.57-8.71 (m, 1H), 8.31 (d, J=5.5 Hz, 2H),
7.31-7.40 (m, 1H), 7.01-7.29 (m, 2H), 3.76-3.95 (m, 1H), 3.59 (d,
J=10.4 Hz, 1H), 3.01 (br. s., 2H), 2.68 (br. s., 4H), 1.95-2.05 (m,
1H), 1.81-1.95 (m, 1 H), 1.59 (br. s., 2H), 1.37-1.53 (m, 1H). MS
(LC/MS) R.T.=0.68; [M+H].sup.+=354.09.
Example 300
(R)--N-(4-(3-Chloro-4-fluoro-phenyl)pyridin-2-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR00963##
[0816]
(R)--N-(4-(3-Chloro-4-fluoro-phenyl)pyridin-2-yl)-4H-1'-azaspiro[ox-
azole-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared from
4-bromopyridin-2-amine, 3-chloro-4-fluorophenylboronic acid, and
1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride
dichloromethane complex by following the general procedures of
Example 28, Steps A-B. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 8.92-9.16 (m, 1H), 8.28 (d, J=5.2 Hz, 1H), 7.88-8.09 (m, 1H),
7.66-7.81 (m, 1H), 7.43-7.63 (m, 1H), 7.19-7.30 (m, 1H), 6.99-7.17
(m, 1H), 3.77-3.94 (m, 1H), 3.53-3.66 (m, 1H), 2.99 (br. s., 2H),
2.67 (br. s., 4H), 1.82-2.03 (m, 2H), 1.52-1.66 (m, 2H), 1.37-1.53
(m, 1H). MS (LC/MS) R.T.=1.65; [M+H].sup.+=387.10.
Example 301
(R)--N-(4-m-Tolylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]o-
ctan]-2-amine
##STR00964##
[0818]
(R)--N-(4-m-Tolylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine was prepared from 4-bromopyridin-2-amine,
m-tolylboronic acid, and
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex by following the general procedures of
Example 28, Steps A-B. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 8.86-9.25 (m, 1H), 8.26 (d, J=5.5 Hz, 1H), 7.45-7.57 (m, 2H),
7.39 (t, J=7.5 Hz, 1H), 7.27 (d, J=7.3 Hz, 1H), 7.18 (d, J=4.3 Hz,
2H), 3.76-3.98 (m, 1H), 3.58 (d, J=10.4 Hz, 1H), 2.99 (d, J=5.8 Hz,
2 H), 2.72-2.85 (m, 2H), 2.67 (t, J=7.6 Hz, 2H), 2.39 (s, 3H),
1.83-2.05 (m, 2H), 1.52-1.64 (m, 2H), 1.36-1.51 (m, 1H). MS (LC/MS)
R.T.=1.54; [M+H].sup.+=349.19.
Example 302
(R)--N-(4-Phenylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]oc-
tan]-2-amine
##STR00965##
[0820]
(R)--N-(4-Phenylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine was prepared from 4-bromopyridin-2-amine by
following the general procedures of Example 28, Steps A-B. .sup.1H
NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.91-9.29 (m, 1H), 8.27 (d,
J=5.5 Hz, 1H), 7.73 (d, J=7.3 Hz, 2H), 7.38-7.58 (m, 3H), 6.92-7.27
(m, 2H), 3.78-3.94 (m, 1H), 3.59 (d, J=10.4 Hz, 1H), 2.91-3.08 (m,
2H), 2.59-2.87 (m, 4H), 1.82-2.05 (m, 2H), 1.59 (br. s., 2H),
1.36-1.53 (m, 1H). MS (LC/MS) R.T.=1.24; [M+H].sup.+=335.15.
Example 303
(R)--N-(4-(1-Methyl-1H-pyrazol-4-yl)pyridin-2-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR00966##
[0822]
(R)--N-(4-(1-Methyl-1H-pyrazol-4-yl)pyridin-2-yl)-4H-1'-azaspiro[ox-
azole-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared from
4-bromopyridin-2-amine,
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole,
and 1,1'-bis(diphenyl phosphino)ferrocene-palladium(II)dichloride
dichloromethane complex by following the general procedures of
Example 28, Steps A-B. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 8.80-9.17 (m, 1H), 8.26 (s, 1H), 8.14 (d, J=5.2 Hz, 1H), 7.94
(br. s., 1H), 7.06 (d, J=4.3 Hz, 2H), 3.88 (s, 4H), 3.56 (d, J=9.8
Hz, 1H), 2.98 (br. s., 2H), 2.60-2.86 (m, 4H), 1.79-2.07 (m, 2H),
1.58 (br. s., 2H), 1.46 (dd, J=9.6, 2.6 Hz, 1H). MS (LC/MS)
R.T.=0.7; [M+H].sup.+=339.18.
Example 304
(R)--N-(4-Thiazol-4-yl)pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2-
.2.2]octan]-2-amine
##STR00967##
[0824]
(R)--N-(4-(Thiazol-4-yl)pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine was prepared from
4-bromopyridin-2-amine, 4-(tributylstannyl)thiazole, and
1,1'-bis(diphenyl phosphino)ferrocene-palladium(II) dichloride
dichloromethane complex by following the general procedures of
Example 28, Steps A-B. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 9.25 (d, J=1.8 Hz, 2H), 8.40 (s, 1H), 8.27 (d, J=5.5 Hz, 1H),
7.23-7.53 (m, 2H), 3.85 (br. s., 1H), 3.58 (d, J=10.4 Hz, 1H), 3.00
(d, J=6.7 Hz, 2H), 2.73-2.85 (m, 2H), 2.68 (t, J=7.8 Hz, 2H),
1.84-2.04 (m, 2H), 1.53-1.68 (m, 2H), 1.47 (d, J=7.0 Hz, 1H). MS
(LC/MS) R.T.=0.78; [M+H].sup.+=342.17.
Example 305
(R)--N-(6-Nitro-3,4'-bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine
##STR00968##
[0826]
(R)--N-(6-Nitro-3,4'-bipyridin-2'-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine was prepared from
4-bromopyridin-2-amine,
2-nitro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine,
and 1,1'-bis(diphenylphosphino) ferrocene-palladium(II) dichloride
dichloromethane complex by following the general procedures of
Example 28, Steps A-B. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 9.06 (br. s., 2H), 8.52-8.67 (m, 1H), 8.31-8.46 (m, 2H),
7.13-7.44 (m, 2H), 3.78-3.94 (m, 1H), 3.60 (d, J=10.1 Hz, 1H), 3.00
(br. s., 2H), 2.59-2.84 (m, 4H), 2.00 (br. s., 2H), 1.59 (br. s.,
2H), 1.38-1.53 (m, 1H). MS (LC/MS) R.T.=0.81;
[M+H].sup.+=381.2.
Example 306
(R)--N-(5-Chloro-6-methylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine
##STR00969##
[0828]
(R)--N-(5-Chloro-6-methylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine was prepared by following the general
procedures of Example 23, Steps A-B using
5-chloro-6-methylpyridin-2-amine as the starting material. .sup.1H
NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.81 (br. s., 1H), 7.59
(br. s., 1H), 6.64 (br. s., 1H), 3.82 (br. s., 1H), 3.56 (d, J=10.4
Hz, 1H), 2.97 (s, 2H), 2.58-2.82 (m, 4H), 2.48 (br. s., 3H), 1.98
(br. s., 1H), 1.81-1.93 (m, 1H), 1.58 (br. s., 2 H), 1.28-1.51 (m,
1H). MS (LC/MS) R.T.=0.89; [M+H].sup.+=307.08.
Example 307
(R)--N-(5-Fluoro-4-methylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine
##STR00970##
[0830]
(R)--N-(5-Fluoro-4-methylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine was prepared by following the general
procedures of Example 23, Steps A-B using
5-fluoro-4-methylpyridin-2-amine as the starting material. .sup.1H
NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.59-8.97 (m, 1H), 8.04 (s,
1H), 6.50-6.95 (m, 1H), 3.79 (br. s., 1H), 3.53 (d, J=10.4 Hz, 1H),
2.96 (br. s., 2H), 2.60-2.84 (m, 4H), 2.21 (s, 3H), 1.95 (br. s.,
2H), 1.52-1.66 (m, 2H), 1.45 (ddd, J=6.9, 2.9, 2.7 Hz, 1H). MS
(LC/MS) R.T.=0.63; [M+H].sup.+=291.12.
Example 308
(R)--N-(3,5-Dichloro-6-methylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00971##
[0832]
(R)--N-(3,5-Dichloro-6-methylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine was prepared by following the
general procedures of Example 23, Steps A-B using
3,5-dichloro-6-methylpyridin-2-amine as the starting material.
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.83 (br. s., 1H),
7.86 (d, J=2.1 Hz, 1H), 3.85 (d, J=9.8 Hz, 1H), 3.59 (d, J=10.1 Hz,
1H), 3.00 (br. s., 1H), 2.73-2.84 (m, 1H), 2.67 (t, J=7.5 Hz, 1H),
2.49 (dd, J=14.5, 2.0 Hz, 6H), 2.01 (br. s., 1H), 1.89 (br. s.,
1H), 1.59 (br. s., 2H), 1.37-1.51 (m, 1H). MS (LC/MS) R.T.=1.07;
[M+H].sup.+=341.06.
Example 309
(R)--N-(4,5-Dimethylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00972##
[0834]
(R)--N-(4,5-Dimethylpyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine was prepared by following the general
procedures of Example 23, Steps A-B using
4,5-dimethylpyridin-2-amine as the starting material. .sup.1H NMR
(500 MHz, DMSO-d.sub.6) .delta. ppm 8.71-9.29 (m, 1H), 7.92 (s,
1H), 6.28-7.02 (m, 1H), 3.80 (br. s., 1H), 3.53 (d, J=10.1 Hz, 1H),
2.87-3.03 (m, 2H), 2.59-2.85 (m, 4H), 2.04-2.26 (m, 6H), 1.80-1.99
(m, 2H), 1.33-1.64 (m, 3H). MS (LC/MS) R.T.=0.67;
[M+H].sup.+=287.20.
Example 310
(R)--N-(5-Methoxypyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]o-
ctan]-2-amine
##STR00973##
[0836]
(R)--N-(5-Methoxypyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine was prepared by following the general
procedures of Example 23, Steps A-B using 5-methoxypyridin-2-amine
as the starting material. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 8.57-9.10 (m, 1H), 7.91 (d, J=3.1 Hz, 1H), 7.30 (dd,
J=8.9, 3.4 Hz, 1H), 6.54-7.04 (m, 1H), 3.72-3.85 (m, 4H), 3.52 (d,
J=10.1 Hz, 1H), 2.88-3.03 (m, 2H), 2.76 (d, J=5.8 Hz, 2H), 2.66 (t,
J=7.6 Hz, 2 H), 1.81-2.02 (m, 2H), 1.51-1.64 (m, 2H), 1.38-1.49 (m,
1H). MS (LC/MS) R.T.=0.46; [M+H].sup.+=289.17.
Example 311
(R)--N-(4-Chloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00974##
[0837] Step A: 4-Chloroisoquinolin-3-amine
##STR00975##
[0839] To isoquinolin-3-amine (2.02 g, 14.0 mmol) in methanol (60
mL) was added N-chlorosuccinimide (2.19 g, 16.4 mmol) at room
temperature. The reaction was stirred at room temperature for 3
hours, then concentrated in vacuo. The crude material was
chromatographed (Biotage: 10-100% ethyl acetate/hexanes) to yield
4-chloroisoquinolin-3-amine (2.1 g, 11.8 mmol, 84% yield) as a
solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.86 (s,
1H), 7.93 (d, J=7.6 Hz, 1H), 7.79 (d, J=8.5 Hz, 1 H), 7.68 (td,
J=6.7, 1.8 Hz, 1H), 7.24-7.35 (m, 1H), 6.32 (br. s., 2H). MS
(LC/MS) R.T.=1.50; [M+H].sup.+=178.96.
Step B:
(R)--N-(4-Chloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00976##
[0841]
(R)--N-(4-Chloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine was prepared by following the general
procedures of Example 267, Steps A-B using
4-chloroisoquinolin-3-amine (from Step A above) as the starting
material. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 8.98-9.10
(m, 1H), 8.57-8.95 (m, 1H), 7.99-8.18 (m, 2H), 7.79 (s, 1H), 7.51
(s, 1H), 3.84 (d, J=9.8 Hz, 1H), 3.35-3.51 (m, 1H), 3.02 (d, J=5.5
Hz, 2H), 2.78 (s, 2H), 2.61-2.73 (m, 2H), 2.00 (s, 2H), 1.55-1.65
(m, 2H), 1.42-1.54 (m, 1H). MS (LC/MS) R.T.=0.92;
[M+H].sup.+=343.09.
Example 312
(R)--N-(7-Chloro-8-fluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00977##
[0842] Step A:
N-(3-Chloro-2-fluorobenzyl)-2,2-diethoxyacetimidamide
##STR00978##
[0844] (3-Chloro-2-fluorophenyl)methanamine (1.0 g, 6.27 mmol) was
added to a solution of methyl 2,2-diethoxyacetimidate (1.12 g, 6.96
mmol) in methanol (10 ml). The mixture was heated at 70 C for 1 h.
The mixture was purified by chromatography (Biotage: 100% ethyl
acetate) to yield
N-(3-chloro-2-fluorobenzyl)-2,2-diethoxyacetimidamide (1.3 g, 4.5
mmol, 65% yield) as a colorless viscous oil. .sup.1H NMR (500 MHz,
CHLOROFORM-d) .delta. ppm 7.23-7.40 (m, 2H), 7.07 (t, J=7.9 Hz,
1H), 4.93 (s, 1H), 4.54 (s, 2H), 3.50-3.74 (m, 4H), 1.18-1.35 (m,
6H). MS (LC/MS) R.T.=1.78; [M+H].sup.+=289.17.
Step B: 7-Chloro-8-fluoroisoquinolin-3-amine
##STR00979##
[0846] To N-(3-chloro-2-fluorobenzyl)-2,2-diethoxyacetimidamide
(0.97 g, 3.36 mmol) was added sulfuric acid (4 mL, 75 mmol). The
reaction was heated to 40.degree. C. for 28 hours. The reaction was
cooled to room temperature and quenched with aq. sodium hydroxide
(.about.15 M) until the reaction mixture was .about.pH 7. The crude
product was extracted with ethyl acetate (2.times.50 mL) and the
organics were dried with MgSO.sub.4, filtered, and concentrated in
vacuo. The crude product was purified by chromatography (Biotage:
100% ethyl acetate to 90/10% ethyl acetate/MeOH) to yield
7-chloro-8-fluoroisoquinolin-3-amine (0.58 g, 2.95 mmol, 88% yield)
as a powder. .sup.1H NMR (500 MHz, DMSO-d.sub.6) d ppm 8.97 (s,
1H), 7.47-7.57 (m, 1H), 7.38-7.45 (m, 1H), 6.67 (s, 1H), 6.33 (s,
2H). MS (LC/MS) R.T.=1.18; [M+H].sup.+=196.95.
Step C:
(R)--N-(7-Chloro-8-fluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole--
5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00980##
[0848]
(R)--N-(7-Chloro-8-fluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine was prepared by following the
general procedures of Example 267, Steps A-B using
7-chloro-8-fluoroisoquinolin-3-amine (from Step B above) as the
starting material. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
9.19 (d, J=2.1 Hz, 1H), 8.30-8.85 (m, 1H), 7.68 (br. s., 2H),
6.93-7.43 (m, 1H), 3.73-4.10 (m, 1H), 3.48-3.70 (m, 1H), 3.02 (br.
s., 2H), 2.59-2.86 (m, 4H), 1.80-2.08 (m, 2 H), 1.60 (br. s., 2H),
1.37-1.53 (m, 1H). MS (LC/MS) R.T.=1.22; [M+H].sup.+=361.06.
Example 313
(R)--N-(1-Methylisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR00981##
[0850]
(R)--N-(1-Methylisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine was prepared by following the general
procedures of Example 312, Steps A-C using 1-phenylethanamine as
the starting material. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.10
(d, J=8.2 Hz, 1H), 7.72 (d, J=8.2 Hz, 1H), 7.60 (t, J=7.5 Hz, 1H),
7.44 (t, J=7.6 Hz, 1H), 7.03-7.33 (m, 1H), 4.01 (d, J=10.1 Hz, 1H),
3.69 (d, J=10.1 Hz, 1H), 3.19-3.31 (m, 2H), 3.12 (d, J=15.0 Hz,
1H), 2.90-3.01 (m, 5H), 2.77-2.90 (m, 2H), 2.17 (br. s., 2H),
1.58-1.87 (m, 3H). MS (LC/MS) R.T.=1.09; [M+H].sup.+=323.16.
Example 314
(R)--N-(6-(4-Fluorophenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00982##
[0852]
(R)--N-(6-(4-Fluorophenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine was prepared by the method of
Example 256, steps A-C, starting from 6-chloropyrimidin-4-amine and
4-fluorophenylboronic acid. .sup.1H NMR (400 MHz, MeOD) .delta. ppm
8.75 (1H, s), 7.93-8.11 (2H, m), 7.06-7.30 (3H, m), 4.03 (1H, d,
J=10.32 Hz), 3.73 (1H, d, J=10.32 Hz), 3.18-3.27 (1H, m), 3.05-3.16
(1H, m), 2.66-3.00 (4H, m), 2.01-2.20 (2H, m), 1.49-1.87 (3H, m).
LC/MS RT=1.53; [M+H].sup.+=354.24.
Example 315
(R)--N-(6-(3-Fluorophenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00983##
[0854]
(R)--N-(6-(3-Fluorophenyl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine was prepared by the method of
Example 256, steps A-C, starting from 6-chloropyrimidin-4-amine and
3-fluorophenylboronic acid. .sup.1H NMR (400 MHz, MeOD) .delta. ppm
8.80 (1H, d, J=1.26 Hz), 7.71-7.85 (2H, m), 7.50 (1H, td, J=7.99,
5.92 Hz), 7.11-7.36 (2H, m), 4.08 (1H, d, J=10.58 Hz), 3.80 (1H, d,
J=10.58 Hz), 3.39-3.48 (1H, m), 3.33 (1H, s), 2.83-3.20 (4H, m),
2.10-2.33 (2H, m), 1.65-1.96 (3H, m). LC/MS RT=1.59;
[M+H].sup.+=354.24.
Example 316
(R)--N-(5-Fluoropyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]-
octan]-2-amine
##STR00984##
[0856]
(R)--N-(5-Fluoropyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine was prepared by the method of Example 231,
steps B-C, starting from 2-chloro-5-fluoropyrimidine. .sup.1H NMR
(400 MHz, MeOD) .delta. ppm 8.50 (2H, d, J=0.76 Hz), 4.03 (1H, d,
J=10.07 Hz), 3.77 (1H, d, J=10.32 Hz), 3.47-3.57 (1H, m), 3.36-3.43
(1H, m), 3.01-3.24 (4H, m), 2.15-2.40 (2H, m), 1.68-2.03 (3H, m).
LC/MS RT=0.30; [M+H].sup.+=278.19.
Example 317
(R)--N-(4-Chloro-5-methoxypyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00985##
[0858]
(R)--N-(4-Chloro-5-methoxypyrimidin-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine was prepared by the method of
Example 231, steps B-C, starting from
2,4-dichloro-5-methoxypyrimidine. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 7.99 (1H, s), 4.13 (1H, d, J=10.83 Hz), 3.91 (4H, d,
J=10.83 Hz), 3.76-3.84 (1H, m), 3.64-3.71 (1H, m), 3.30-3.51 (4H,
m), 2.43-2.52 (1H, m), 2.37 (1H, d, J=3.27 Hz), 2.09 (1H, dd,
J=9.32, 4.78 Hz), 1.88-2.03 (2H, m). LC/MS RT=0.30;
[M+H].sup.+=278.19.
Example 318
(R)--N-(6,8-Difluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine
##STR00986##
[0860]
(R)--N-(6,8-Difluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine was prepared by the method of Example
279, steps A-D, starting from (3,5-difluorophenyl)methanamine MS
(LC/MS) R.T.=1.42; [M+H].sup.+=345.26.
Example 319
(R)--N-(6,7-Difluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine
##STR00987##
[0862]
(R)--N-(6,7-Difluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine was prepared by the method of Example
279, steps A-D, starting from (3,4-difluorophenyl)methanamine.
.sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.97 (1H, s), 7.73-7.91
(1H, m), 7.58 (1H, dd, J=11.14, 7.78 Hz), 7.30 (1H, s), 4.01 (1H,
d, J=10.07 Hz), 3.71 (1H, d, J=10.07 Hz), 3.31 (1H, br. s.),
3.14-3.24 (1H, m), 3.03 (2H, d, J=7.32 Hz), 2.83-2.98 (2H, m), 2.20
(2H, br. s.), 1.61-1.91 (3H, m). MS (LC/MS) R.T.=1.45;
[M+H].sup.+=345.26.
Example 320
(R)--N-(5,8-Difluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[-
2.2.2]octan]-2-amine (
##STR00988##
[0864]
(R)--N-(5,8-Difluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine was prepared by the method of Example
279, steps A-D, starting from (2,5-difluorophenyl)methanamine.
.sup.1H NMR (500 MHz, MeOD) .delta. ppm 9.25 (1H, s), 7.43 (1H, br.
s.), 7.20-7.34 (1H, m), 6.91-7.12 (1H, m), 4.04 (1H, d, J=10.07
Hz), 3.74 (1H, d, J=10.07 Hz), 3.30 (1H, br. s.), 3.13-3.23 (1H,
m), 3.03 (2H, d, J=7.63 Hz), 2.82-2.96 (2H, m), 2.21 (2H, br. s.),
1.58-1.92 (3H, m). MS (LC/MS) R.T.=1.37; [M+H].sup.+=345.33.
Example 321
(R)--N-(7-Fluoroquinazolin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo
octan]-2-amine
##STR00989##
[0866]
(R)--N-(7-Fluoroquinazolin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine was prepared by the method of Example 239,
steps A-B, starting from 7-fluoroquinazolin-2-amine .sup.1H NMR
(400 MHz, MeOD) .delta. ppm 9.22 (1H, s), 7.95 (1 H, dd, J=8.81,
6.04 Hz), 7.47 (1H, dd, J=10.45, 2.14 Hz), 7.24 (1H, td, J=8.75,
2.39 Hz), 4.07 (1H, d, J=10.07 Hz), 3.77 (1H, d, J=10.07 Hz), 3.25
(1H, s), 3.07-3.17 (1H, m), 2.89-3.02 (2H, m), 2.75-2.88 (2H, m),
2.04-2.26 (2H, m), 1.50-1.85 (3H, m). MS (LC/MS) R.T.=1.06;
[M+H].sup.+=328.33.
Example 322
(R)--N-(5-Chloroquinazolin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00990##
[0868]
(R)--N-(5-Chloroquinazolin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine was prepared by the method of Example 239,
steps A-B, starting from 5-chloroquinazolin-2-amine. .sup.1H NMR
(400 MHz, MeOD) .delta. ppm 9.53 (1H, s), 7.62-7.84 (2H, m), 7.47
(1H, dd, J=6.80, 1.51 Hz), 4.09 (1H, d, J=10.32 Hz), 3.80 (1H, d,
J=10.32 Hz), 3.36 (1H, s), 3.15-3.24 (1H, m), 3.00 (2H, t, J=7.68
Hz), 2.79-2.95 (2H, m), 2.05-2.32 (2H, m), 1.53-1.87 (3H, m). MS
(LC/MS) R.T.=1.36; [M+H].sup.+=344.29
Example 323
(R)--N-(5-Fluoroquinazolin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine
##STR00991##
[0870]
(R)--N-(5-Fluoroquinazolin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine was prepared by the method of Example 239,
steps A-B, starting from 5-fluoroquinazolin-2-amine .sup.1H NMR
(400 MHz, MeOD) .delta. ppm 9.46 (1H, s), 7.77 (1 H, td, J=8.18,
6.30 Hz), 7.63 (1H, d, J=8.56 Hz), 7.00-7.20 (1H, m), 4.08 (1H, d,
J=10.07 Hz), 3.77 (1H, d, J=10.07 Hz), 3.25 (1H, s), 3.07-3.17 (1H,
m), 2.94 (2H, t, J=7.68 Hz), 2.72-2.88 (2H, m), 2.05-2.21 (2H, m),
1.48-1.83 (3H, m). MS (LC/MS) R.T.=1.10; [M+H].sup.+=328.33
Example 324
(R)--N-(7-Fluoro-4-methylquinazolin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR00992##
[0872]
(R)--N-(7-Fluoro-4-methylquinazolin-2-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine was prepared by the method of
Example 239, steps A-B, starting from
7-fluoro-4-methylquinazolin-2-amine .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.13 (1H, dd, J=9.07, 6.04 Hz), 7.46 (1H, dd, J=10.45,
2.64 Hz), 7.22 (1H, td, J=8.81, 2.77 Hz), 4.06 (1H, d, J=10.07 Hz),
3.75 (1H, d, J=10.07 Hz), 3.23 (1H, s), 3.06-3.15 (1H, m), 2.94
(2H, t, J=7.68 Hz), 2.73-2.88 (5H, m), 2.06-2.24 (2 H, m),
1.51-1.86 (3H, m). MS (LC/MS) R.T.=1.25; [M+H].sup.+=342.29.
Example 325
(R)--N-(6-Fluoro-1-methylisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00993##
[0874]
(R)--N-(6-Fluoro-1-methylisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine was prepared by the method of
Example 279, steps A-D, starting from
(S)-1-(4-fluorophenyl)ethanamine .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.23 (1H, dd, J=9.16, 5.49 Hz), 7.44 (1H, dd, J=9.92,
2.59 Hz), 7.22-7.38 (2 H, m), 4.17 (1H, d, J=10.99 Hz), 3.94 (1H,
d, J=10.68 Hz), 3.71-3.79 (1H, m), 3.61-3.69 (1H, m), 3.39-3.49
(1H, m), 3.33-3.39 (3H, m), 2.95 (3H, s), 2.49 (1 H, br. s.), 2.37
(1H, tt, J=10.07, 3.36 Hz), 2.05-2.16 (1H, m), 1.93-2.05 (2H, m).
MS (LC/MS) R.T.=1.47; [M+H].sup.+=341.25
Example 326
(R)--N-(6-Chloro-1-methylisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00994##
[0876]
(R)--N-(6-Chloro-1-methylisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine was prepared by the method of
Example 279, steps A-D, starting from 1-(4-chlorophenyl)ethanamine
.sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.23 (1H, dd, J=9.16, 5.49
Hz), 7.44 (1H, dd, J=9.92, 2.59 Hz), 7.22-7.38 (2H, m), 4.17 (1H,
d, J=10.99 Hz), 3.94 (1H, d, J=10.68 Hz), 3.71-3.79 (1H, m),
3.61-3.69 (1H, m), 3.39-3.49 (1H, m), 3.33-3.39 (3H, m), 2.95 (3H,
s), 2.49 (1H, br. s.), 2.37 (1H, tt, J=10.07, 3.36 Hz), 2.05-2.16
(1H, m), 1.93-2.05 (2H, m). MS (LC/MS) R.T.=1.79;
[M+H].sup.+=357.28
Example 327
(R)--N-(7-Fluoro-1-methylisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR00995##
[0878]
(R)--N-(7-Fluoro-1-methylisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine was prepared by the method of
Example 279, steps A-D, starting from 1-(3-fluorophenyl)ethanamine
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 7.55-7.81 (2H, m), 7.38
(1H, t, J=7.55 Hz), 7.16 (1H, br. s.), 3.95 (1H, d, J=9.82 Hz),
3.64 (1H, d, J=9.82 Hz), 3.20 (1H, d, J=14.86 Hz), 3.01-3.13 (1H,
m), 2.63-2.97 (7H, m), 2.11 (2H, br. s.), 1.41-1.81 (3H, m). MS
(LC/MS) R.T.=1.63; [M+H].sup.+=341.32.
Example 328
(R)--N-(5,7-Difluoro-1-methylisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octan]-2-amine
##STR00996##
[0880]
(R)--N-(5,7-Difluoro-1-methylisoquinolin-3-yl)-4H-1'-azaspiro[oxazo-
le-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared by the method of
Example 279, steps A-D, starting from
1-(3,5-difluorophenyl)ethanamine .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 7.78 (1H, d, J=9.77 Hz), 7.39-7.55 (2H, m), 4.31 (1H,
d, J=10.99 Hz), 4.11 (1H, d, J=10.99 Hz), 3.88-3.97 (1H, m),
3.79-3.86 (1H, m), 3.55 (1H, t, J=11.90 Hz), 3.37-3.47 (3H, m),
3.00 (3H, s), 2.64 (1H, br. s.), 2.32-2.45 (1H, m), 2.18 (1H, dddd,
J=11.71, 7.13, 6.94, 6.56 Hz), 1.99-2.13 (2H, m). MS (LC/MS)
R.T.=1.67; [M+H].sup.+=359.22.
Example 329
(R)--N-(6-(6-(Methylthio)pyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazo-
le-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR00997##
[0882]
(R)--N-(6-(6-(Methylthio)pyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspir-
o[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared by the
method of Example 261, steps A-B, starting from
6-(6-fluoropyridin-3-yl)pyrimidin-4-amine .sup.1H NMR (400 MHz,
MeOD) .delta. ppm 9.02 (1H, d, J=2.52 Hz), 8.78 (1H, s), 8.20 (1H,
dd, J=8.44, 2.39 Hz), 7.36 (1H, d, J=8.56 Hz), 7.26 (1H, br. s.),
4.06 (1H, d, J=10.58 Hz), 3.77 (1H, d, J=10.32 Hz), 3.34 (1H, s),
3.14-3.22 (1H, m), 2.82-3.08 (4H, m), 2.58 (3H, s), 2.08-2.24 (2H,
m), 1.60-1.88 (3H, m). MS (LC/MS) R.T.=3.09; [M+H].sup.+=383.2.
Example A 330
(R)--N-(6-(2-Fluoropyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine
##STR00998##
[0883] Step A: 6-(2-Fluoropyridin-3-yl)pyrimidin-4-amine
##STR00999##
[0885] A mixture of 6-chloropyrimidin-4-amine (1.5 g, 11.58 mmol),
2-fluoropyridin-3-ylboronic acid (2.039 g, 14.47 mmol), and
Na.sub.2CO.sub.3 (3.68 g, 34.7 mmol) was suspended in a mixture of
dioxane (15 mL)/EtOH (2 mL)/water (3 mL). The mixture was heated in
the microwave synthesizer at 125.degree. C. for 20 min,
concentrated, and purified on a silica gel cartridge using 10-60%
ethyl acetate in hexanes, then 5-25% 9:1 methanol:ammonium
hydroxide in ethyl acetate to give an off-white solid (1.5 g, 7 89
mmol, 68%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 8.57
(1H, ddd, J=9.95, 7.68, 2.01 Hz), 8.48 (1H, d, J=1.26 Hz), 8.33
(1H, td, J=3.02, 2.01 Hz), 7.51 (1H, ddd, J=7.37, 4.97, 2.01 Hz),
7.12 (2H, br. s.), 6.96 (1H, d, J=1.26 Hz). MS (LC/MS) R.T.=0.43;
[M+H].sup.+=191.15.
Step B: 4-(2-Fluoropyridin-3-yl)-6-isothiocyanatopyrimidine
##STR01000##
[0887] To a solution of 6-(2-fluoropyridin-3-yl)pyrimidin-4-amine
(0.3 g, 1.577 mmol) in DMF was added NaH (0.126 g, 3.15 mmol).
Stirring was continued for 1.5 h and
1,1'-thiocarbonyldipyridin-2(1H)-one (0.366 g, 1.577 mmol) was
added. The mixture was stirred at rt for 2 h and purified on silica
gel using 10-15% ethyl acetate in hexanes to give an orange solid
(0.133 g, 0.573 mmol, 36%). MS (LC/MS) R.T.=2.48;
[M+H].sup.+=233.08.
Step C:
(S)-1-(6-(2-Fluoropyridin-3-yl)pyrimidin-4-yl)-3-((3-hydroxyquinuc-
lidin-3-yl)methyl)thiourea
##STR01001##
[0889] To (4-(2-fluoropyridin-3-yl)-6-isothiocyanatopyrimidine
(0.18 g, 0.775 mmol) in N,N-dimethylformamide (20 mL) was added
Cs.sub.2CO.sub.3 (0.631 g, 1.938 mmol) and
(S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.178 g, 0.775
mmol). The suspension was stirred at room temperature for 18 h and
concentrated. The residue was purified on silica gel using 0-10%
9.5:0.5 methanol:ammonium hydroxide in ethyl acetate to give a
yellow solid (0.2 g, 0.515 mmol, 66%). MS (LC/MS) R.T.=1.82;
[M+H].sup.+=389.27.
Step D:
(R)--N-(6-(2-Fluoropyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxa-
zole-5,3'-bicyclo[2.2.2]octan]-2-amine)
##STR01002##
[0891] To suspension of
((S)-1-(6-(2-fluoropyridin-3-yl)pyrimidin-4-yl)-3-((3-hydroxyquinuclidin--
3-yl)methyl)thiourea (0.2 g, 0.515 mmol) in N,N-dimethylformamide
(20 mL) was added Cs.sub.2CO.sub.3 (0.168 g, 0.515 mmol) and DIC
(0.241 mL, 1.545 mmol). The suspension was stirred at room
temperature for 18 h, then at 60.degree. C. for 3 h. The mixture
was concentrated and the residue was purified on silica gel using
2-7% 9.5:0.5 methanol:ammonium hydroxide in ethyl acetate to afford
(R)--N-(6-(2-fluoropyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine as a pale yellow solid (0.029 g,
0.081 mmol, 16%). .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.83 (1H,
d, J=1.26 Hz), 8.55 (1H, ddd, J=9.76, 7.62, 2.01 Hz), 8.29 (1H, dt,
J=3.02, 1.51 Hz), 7.46 (1H, ddd, J=7.37, 5.10, 1.89 Hz), 7.33 (1H,
s), 4.06 (1H, d, J=10.32 Hz), 3.76 (1H, d, J=10.32 Hz), 3.25 (1H,
s), 3.09-3.19 (1H, m), 2.95 (2H, t, J=7.68 Hz), 2.74-2.91 (2H, m),
2.04-2.26 (2 H, m), 1.55-1.87 (3H, m). MS (LC/MS) R.T.=1.24;
[M+H].sup.+=355.28.
Example 331
(R)--N-(6-(2-Fluoropyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine
##STR01003##
[0893]
(R)--N-(6-(6-Fluoropyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxaz-
ole-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared by the method of
Example 330, steps A-D, starting from 6-fluoropyridin-3-ylboronic
acid. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.88 (1H, d, J=2.52
Hz), 8.82 (1H, s), 8.51-8.60 (1H, m), 7.25 (1H, s), 7.19 (1H, dd,
J=8.56, 2.52 Hz), 4.06 (1H, d, J=10.32 Hz), 3.76 (1H, d, J=10.32
Hz), 3.22 (1H, s), 3.07-3.14 (1H, m), 2.86-2.98 (2H, m), 2.70-2.87
(2H, m), 2.03-2.20 (2H, m), 1.54-1.87 (3H, m). MS (LC/MS)
R.T.=1.21; [M+H].sup.+=355.28.
Example 332
(R)--N-(6-(5-Fluoropyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine
##STR01004##
[0895]
(R)--N-(6-(5-Fluoropyridin-3-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxaz-
ole-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared by the method of
Example 330, steps A-D, starting from 5-fluoropyridin-3-ylboronic
acid. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.06 (1H, d, J=1.26
Hz), 8.81 (1H, d, J=1.01 Hz), 8.56 (1H, d, J=2.77 Hz), 8.26 (1H,
dt, J=9.82, 2.27 Hz), 7.30 (1H, br. s.), 4.06 (1H, d, J=10.32 Hz),
3.75 (1 H, d, J=10.32 Hz), 3.16-3.26 (1H, m), 3.04-3.15 (1H, m),
2.86-3.00 (2H, m), 2.81 (2H, t, J=7.30 Hz), 2.01-2.22 (2H, m),
1.52-1.84 (3H, m). MS (LC/MS) R.T.=1.20; [M+H].sup.+=355.28.
Example 333
(R)--N-(6-(1-Methyl-1H-pyrazol-5-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-
-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01005##
[0897]
((R)--N-(6-(1-Methyl-1H-pyrazol-5-yl)pyrimidin-4-yl)-4H-1'-azaspiro-
[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine was prepared by the
method of Example 330, steps A-D, starting from
1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole.
.sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.89 (1H, s), 7.54 (1H, d,
J=2.14 Hz), 7.28 (1H, br. s.), 6.84 (1H, d, J=2.14 Hz), 4.23 (3H,
s), 4.18 (1H, d, J=10.99 Hz), 3.98 (1H, d, J=10.68 Hz), 3.81-3.88
(1H, m), 3.72-3.80 (1H, m), 3.53 (1H, t, J=11.75 Hz), 3.34-3.46
(3H, m), 2.53 (1H, br. s.), 2.33-2.45 (1H, m), 2.10-2.22 (1H, m),
1.97-2.12 (2H, m). MS (LC/MS) R.T.=1.15; [M+H].sup.+=340.29.
Example 334
(R)--N-(6-(Pyrazin-2-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine
##STR01006##
[0898] Step A:
6[Bis(tert-butoxycarbonyl)amino]-4-chloropyrimidine
##STR01007##
[0900] 6-Chloropyrimidin-4-amine (4.1 g, 31.6 mmol) was suspended
in acetonitrile. DMAP (0.966 g, 7.91 mmol) and di-tert-butyl
dicarbonate (14.74 g, 67.5 mmol) were added and the mixture was
stirred at rt for 3 days. The solvent was evaporated and the
residue was purified by silica gel using 0-15% ethyl acetate in
hexanes to give 6[bis(tert-butoxycarbonyl)amino]-4-chloropyrimidine
as a white solid (6.27 g, 19.01 mmol, 60%). .sup.1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 8.69 (1H, d, J=0.76 Hz), 7.88 (1H, d,
J=1.01 Hz), 1.58 (18H, s).
Step B:
6[Bis(tert-butoxycarbonyl)amino]-4-(2'-pyrazinyl)pyrimidine
##STR01008##
[0902] 6[Bis(tert-butoxycarbonyl)amino]-4-chloropyrimidine (1.608
g, 4.88 mmol), 2-(tributylstannyl)pyrazine (1.5 g, 4.06 mmol) and
Pd(Ph.sub.3P).sub.4 (0.225 g, 0.195 mmol) were combined in toluene,
flushed with nitrogen and heated under reflux for 17 h, cooled to
rt, concentrated, and purified on silica gel using 10% ethyl
acetate in hexanes, then 20-50% ethyl acetate in hexanes to give a
yellow solid (0.94 g, 2.507 mmol, 62%). .sup.1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 9.67 (1H, d, J=1.01 Hz), 9.05 (1H, d,
J=1.26 Hz), 8.60-8.77 (3H, m), 1.59 (18H, s).
Step C: 6-(Pyrazin-2-yl)pyrimidin-4-amine
##STR01009##
[0904] Trifluoroacetic acid (2.77 mL, 36.0 mmol) was added to a
solution of
6[bis(tert-butoxycarbonyl)amino]-4-(2'-pyrazinyl)pyrimidine (1.344
g, 3.60 mmol) in dichloromethane at rt. The mixture was stirred at
rt for 18 h and concentrated. The residue was taken up in ethyl
acetate and saturated NaHCO.sub.3 was carefully added. The organic
layer was isolated, dried over Na.sub.2SO.sub.4 and concentrated.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.44 (1H, d, J=1.01
Hz), 8.63-8.86 (2H, m), 8.51 (1H, d, J=1.01 Hz), 7.38 (1H, d,
J=1.26 Hz), 7.17 (2H, br. s.). MS (LC/MS) R.T.=0.37;
[M+H].sup.+=174.23.
Step D: Dimethyl
6-(pyrazin-2-yl)pyrimidin-4-ylcarbonimidodithioate
##STR01010##
[0906] To a solution of 6-(pyrazin-2-yl)pyrimidin-4-amine (0.2 g,
1.155 mmol) in DMF was added NaOH (0.115 mL, 2.310 mmol), CS.sub.2
(0.174 mL, 2.89 mmol), NaOH (0.115 mL, 2.310 mmol) and Met (0.181
mL, 2.89 mmol) at 15 min intervals. Stirring was continued for 1.5
h and the mixture was poured into water. The orange solid was
separated, washed with water, dried to give a bright yellow solid
(0.14 g, 0.505 mmol, 44%). MS (LC/MS) R.T.=3.19;
[M+H].sup.+=278.19.
Step E:
(R)--N-(6-(Pyrazin-2-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3-
'-bicyclo[2.2.2]octan]-2-amine
##STR01011##
[0908] A mixture of dimethyl
6-(pyrazin-2-yl)pyrimidin-4-ylcarbonimidodithioate (0.139 g, 0.501
mmol), Cs.sub.2CO.sub.3 (0.408 g, 1.253 mmol) and
(S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (0.115 g, 0.501
mmol) in N,N-dimethylformamide (10 ml) was heated at 80.degree. C.
for 3 h. The mixture was concentrated and purified on silica gel
using 0-10% 9.5:0.5 methanol:ammonium hydroxide in ethyl acetate to
give
(R)--N-(6-(pyrazin-2-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyc-
lo[2.2.2]octan]-2-amine as an off-white solid (0.066 g, 0.194 mmol,
39%). .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.46 (1H, d, J=1.51
Hz), 8.83 (1H, d, J=1.26 Hz), 8.67-8.72 (1H, m), 8.66 (1H, d,
J=2.52 Hz), 7.77 (1H, br. s.), 4.05 (1H, d, J=10.32 Hz), 3.75 (1H,
d, J=10.32 Hz), 3.18-3.26 (1H, m), 3.04-3.15 (1H, m), 2.87-3.00
(2H, m), 2.70-2.85 (2H, m), 2.04-2.19 (2H, m), 1.54-1.83 (3H, m).
MS (LC/MS) R.T.=0.97; [M+H].sup.+=338.29.
Example 335
(2R)--N-(6,7-Dimethyl-3-isoquinolinyl)-4'H-spiro[4-azabicyclo[2.2.2]octane-
-2,5'-[1,3]oxazol]-2'-amine
##STR01012##
[0909] Step A: 6,7-Dimethylisoquinolin-3-amine
##STR01013##
[0911] To a solution of methyl 2,2-diethoxyacetimidate (1.50 g,
9.32 mmol) in methanol (4.9 mL) was added
(3,4-dimethylphenyl)methanamine (1.20 g, 8.88 mmol) dropwise at
ambient temperature. The reaction flask was then placed into a
preheated oil-bath and stirred at 70.degree. C. for 16 h; after
which time, the flask was removed from the oil-bath and allowed to
cool to ambient temperature. The volatiles were removed under
reduced pressure and the crude material was added dropwise to
sulfuric acid (19.7 mL) at ambient temperature. The reaction
mixture was stirred for 72 h, then the flask was placed into an
ice-water bath, diluted with water (50 mL), and slowly neutralized
to pH=10 with sodium hydroxide (10 N). As the reaction mixture
became basic, an orange precipitate formed. This precipitate was
filtered, washed with water, and dried to afford a mixture (2:1) of
6,7-dimethylisoquinolin-3-amine and 5,6-dimethylisoquinolin-3-amine
(1.68 g, 9.75 mmol, >100%). MS (LC/MS) R.T.=0.32;
[M+H].sup.+=173.13.
Step B: 3-Isothiocyanato-6,7-dimethylisoquinoline
##STR01014##
[0913] To a solution of 1,1'-thiocarbonyldipyridin-2(1H)-one (1.35
g, 5.81 mmol) in dichloromethane (19 mL) at ambient temperature was
added a mixture (2:1) of 6,7-dimethylisoquinolin-3-amine and
5,6-dimethylisoquinolin-3-amine (1.00 g, 5.81 mmol). The reaction
mixture was placed into a preheated oil-bath and stirred at
40.degree. C. for 18 h, then cooled, concentrated, and purified by
silica gel chromatography (10-35% ethyl acetate in hexanes) to
afford a mixture (2:1) of 3-isothiocyanato-6,7-dimethylisoquinoline
and 3-isothiocyanato-5,6-dimethylisoquinoline (186 mg, 0.869 mmol,
15%) as a yellow solid. MS (LC/MS) R.T.=2.06;
[M+H].sup.+=215.1.
Step C:
(2R)--N-(6,7-Dimethyl-3-isoquinolinyl)-4'H-spiro[4-azabicyclo[2.2.-
2]octane-2,5'-[1,3]oxazol]-2'-amine
##STR01015##
[0915] To (S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (99
mg, 0.43 mmol) in N,N-dimethylformamide (1.4 mL) was added
triethylamine (0.181 mL, 1.30 mmol) and a mixture (2:1) of
3-isothiocyanato-6,7-dimethylisoquinoline and
3-isothiocyanato-5,6-dimethylisoquinoline (93 mg, 0.43 mmol). The
suspension was placed into a preheated oil-bath and stirred at
80.degree. C. for 2 h and 30 min. N,N'-Diisopropylcarbodiimide
(0.270 mL, 1.74 mmol) was then added and the mixture was stirred at
80.degree. C. for 16 h. The mixture was concentrated and purified
by silica gel chromatography (0-40% 9:1 methanol:ammonium hydroxide
in chloroform) followed by purification by reverse phase
preparatory HPLC (0-40% TFA-methanol-water). The product fractions
were combined and concentrated in vacuo to afford a mixture (2:1)
of
(2R)--N-(6,7-dimethyl-3-isoquinolinyl)-4'H-spiro[4-azabicyclo[2.-
2.2]octane-2,5'-[1,3]oxazol]-2'-amine and
(2R)--N-(5,6-dimethyl-3-isoquinolinyl)-4'H-spiro[4-azabicyclo[2.2.2]octan-
e-2,5'-[1,3]oxazol]-2'-amine as the trifluoroacetic acid salts. The
two regioisomers were separated using a Chiralpak AD-H
(4.6.times.250 mm, 5 .mu.m) column with a mobile phase consisting
of 40% methanol (0.1% DEA) in CO.sub.2, detected at 215 nM, to
yield
(2R)--N-(6,7-dimethyl-3-isoquinolinyl)-4'H-spiro[4-azabicyclo[2.2.2]octan-
e-2,5'-[1,3]oxazol]-2'-amine as a white solid (5.0 mg, 0.015 mmol,
3% yield). 1H NMR (400 MHz, MeOD) .delta. ppm 8.87 (1H, s), 7.90
(1H, s), 7.67 (1H, s), 7.50 (1H, s), 3.97 (1H, d, J=10.0 Hz), 3.65
(1H, d, J=10.0 Hz), 3.19-3.27 (1H, m), 3.04-3.17 (1H, m), 2.94 (2H,
d, J=7.5 Hz), 2.76-2.90 (2H, m), 2.38-2.50 (6H, m), 2.11-2.21 (2H,
m), 1.57-1.85 (3H, m). MS (LC/MS) R.T.=0.92;
[M+H].sup.+=337.22.
Example 336
(2R)--N-(6-Methyl-1,3-benzoxazol-2-yl)-4'H-spiro[4-azabicyclo[2.2.2]octane-
-2,5'-[1,3]oxazol]-2'-amine
##STR01016##
[0916] Step A: 6-Methylbenzo[d]oxazol-2-amine
##STR01017##
[0918] An oven-dried, round-bottomed flask was charged with
di(1H-imidazol-1-yl)methanimine (1.40 g, 8.69 mmol),
2-amino-5-methylphenol (713 mg, 5.79 mmol) and anhydrous THF (20
ml) at ambient temperature. The resulting suspension was refluxed
under N.sub.2 (g) for 2 h. The solvent was removed in vacuo and the
residue was purified by silica gel chromatography (0-30% 9:1
methanol:ammonium hydroxide-chloroform) to afford
benzo[d]oxazol-2-amine (792 mg, 5.35 mmol, 92% yield), as a grey
solid. 1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 7.19-7.34 (1H,
m), 7.11 (1H, s), 7.01 (1H, d, J=7.8 Hz), 5.60 (2H, br. s.), 2.43
(3H, s). MS (LC/MS) R.T.=0.89; [M+H].sup.+=149.09.
Step B: Dimethyl
6-methylbenzo[d]oxazol-2-ylcarbonimidodithioate
##STR01018##
[0920] To a suspension of 6-methylbenzo[d]oxazol-2-amine (200 mg,
1.35 mmol) in DMF (1.4 mL) was added 20.0 M sodium hydroxide (135
.mu.L, 2.70 mmol). The mixture was allowed to stir for 10 min at
room temperature before carbon disulfide was added (203 .mu.L, 3.37
mmol) and the mixture was stirred for 10 min. An additional portion
of 20.0 M sodium hydroxide (135 .mu.L, 2.70 mmol) was added and the
mixture was again stirred for 10 min. Finally, iodomethane (203
.mu.L, 3.24 mmol) was added dropwise. After completion of the
addition, the mixture was stirred for 15 min, at which time a
voluminous precipitate had formed. The mixture was poured into
water and the solids were collected by filtration, washed with
water, and dried to afford dimethyl
6-methylbenzo[d]oxazol-2-ylcarbonimidodithioate (289 mg, 1.14 mmol,
85% yield) as a brown solid. MS (LC/MS) R.T.=1.88;
[M+H].sup.+=252.95.
Step C:
(2R)--N-(6-Methyl-1,3-benzoxazol-2-yl)-4'H-spiro[4-azabicyclo[2.2.-
2]octane-2,5'-[1,3]oxazol]-2'-amine
##STR01019##
[0922] A 10 ml vial was charged with
(S)-3-(aminomethyl)quinuclidin-3-ol.HCl (38.2 mg, 0.198 mmol, DMF
(2 mL), Cs.sub.2CO.sub.3 (129 mg, 0.396 mmol) and dimethyl
6-methylbenzo[d]oxazol-2-ylcarbonimidodithioate (50 mg, 0.198 mmol)
at ambient temperature. The resulting suspension was stirred for 1
h before it was diluted with methanol, and purified by reverse
phase preparatory HPLC (0-100% TFA-methanol-water) to afford
(2R)--N-(6-methyl-1,3-benzoxazol-2-yl)-4'H-spiro[4-azabicyclo[2.2.2]octan-
e-2,5'-[1,3]oxazol]-2'-amine as the trifluoroacetic acid salt (55.3
mg, 0.123 mmol, 62% yield) as a tan gum. 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 9.89 (br. s., 1H), 9.15 (br. s., 1H),
7.33 (d, J=8.03 Hz, 1H), 7.31 (s, 1H), 7.06 (d, J=8.78 Hz, 1H),
4.00 (d, J=8.0 Hz, 1H), 3.88 (d, J=10.29 Hz, 1H), 3.64-3.80 (m,
2H), 3.34-3.44 (m, 1H), 3.20-3.34 (m, 3H), 2.44 (br. s., 1H), 2.40
(s, 3H), 2.08-2.20 (m, 1H), 1.80-2.03 (m, 3H). MS (LC/MS)
R.T.=0.367, [M+H].sup.+=313.2.
Example 337
(2R)--N-(6-Methoxy-1,3-benzoxazol-2-yl)-4'H-spiro[4-azabicyclo[2.2.2]octan-
e-2,5'-[1,3]oxazol]-2'-amine
##STR01020##
[0923] Step A: 6-Methoxybenzo[d]oxazol-2-amine
##STR01021##
[0925] An oven-dried, round-bottomed flask was charged with
di(1H-imidazol-1-yl)methanimine (500 mg, 3.10 mmol),
2-amino-5-methoxyphenol-HCl (365 mg, 2.07 mmol), triethylamine (288
.mu.L, 2.07 mmol), and anhydrous THF (20 ml) at ambient
temperature. The resulting suspension was refluxed under N.sub.2
(g) for 20 h to give complete conversion based on LC/MS. The
solvent was removed in vacuo and the residue was purified by silica
gel chromatography (30-80% ethyl acetate in hexanes) to afford
6-methoxybenzo[d]oxazol-2-amine (307 mg, 1.87 mmol, 90% yield), as
a brown solid. 1H NMR (500 MHz, CHLOROFORM-d) .delta. ppm 7.28 (1H,
d, J=9.5 Hz), 6.92 (1H, d, J=2.4 Hz), 6.80 (1H, dd, J=8.5, 2.1 Hz),
5.28 (2H, br. s.). MS (LC/MS) R.T.=1.51; [M+H].sup.+=165.00.
Step B: Dimethyl
6-methoxybenzo[d]oxazol-2-ylcarbonimidodithioate
##STR01022##
[0927] To a suspension of 6-methoxybenzo[d]oxazol-2-amine (238 mg,
1.45 mmol) in DMF (2.0 mL) was added 20.0 M sodium hydroxide (145
.mu.L, 2.90 mmol). The mixture was allowed to stir for 10 min at
room temperature before carbon disulfide was added (219 .mu.L, 3.62
mmol) and the mixture was stirred for 10 min. An additional portion
of 20.0 M sodium hydroxide (145 .mu.L, 2.90 mmol) was added and the
mixture was again stirred for 10 min. Finally, iodomethane (218
.mu.L, 3.48 mmol) was added dropwise. The mixture was stirred for
an additional 15 min, by which time a voluminous precipitate had
formed. The mixture was poured into water and the solids were
collected by filtration, washed with water, and dried to afford
dimethyl 6-methoxybenzo[d]oxazol-2-ylcarbonimidodithioate. The
material was used without characterization.
Step C:
(2R)--N-(6-Methoxy-1,3-benzoxazol-2-yl)-4'H-spiro[4-azabicyclo[2.2-
.2]octane-2,5'-[1,3]oxazol]-2'-amine
##STR01023##
[0929] A 10 ml vial was charged with
(S)-3-(aminomethyl)quinuclidin-3-ol.HCl (24.5 mg, 0.127 mmol), DMF
(2 mL), Cs.sub.2CO.sub.3 (83 mg, 0.25 mmol), and dimethyl
6-methoxybenzo[d]oxazol-2-ylcarbonimidodithioate (34 mg, 0.13 mmol)
at ambient temperature. The resulting suspension was stirred at
ambient temperature for 1 h, diluted with methanol and purified by
reverse phase preparatory HPLC (0-100% TFA-methanol-water) to
afford
(R)--N-(6-methoxybenzo[d]oxazol-2-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine as the trifluoroacetic acid salt (7.2 mg,
0.014 mmol, 11% yield) as a tan gum. 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 9.95 (br. s., 1H), 9.10 (br. s., 1H), 7.34 (d, J=8.78
Hz, 1H), 7.17 (d, J=2.51 Hz, 1H), 6.84 (dd, J=8.53, 2.51 Hz, 1H),
3.99 (d, J=10.54 Hz, 1H), 3.87 (d, J=10.54 Hz, 1H), 3.79 (s, 3H),
3.63-3.78 (m, 2H), 3.34-3.44 (m, 1H), 3.20-3.32 (m, 3H), 2.43 (m,
1H), 2.09-2.19 (m, 1H), 1.91-2.02 (m, 1H), 1.80-1.91 (m, 2 H). MS
(LC/MS) R.T.=0.867, [M+H].sup.+=329.28.
Examples 338 A and 338 B
(R)--N-(5-chloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine (A) and
(R)--N-(7-chloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2-
.2]octan]-2-amine (B)
##STR01024##
[0930] Step A: N-(3-Chlorobenzyl)-2,2-diethoxyacetimidamide
##STR01025##
[0932] In a nitrogen-flushed sealed tube was placed methyl
2,2-diethoxyacetimidate (2 g, 12.41 mmol) in methanol (6 mL) and
(3-chlorophenyl)methanamine (1.444 mL, 11.82 mmol). The tube was
capped and heated to 70.degree. C. After 18 hours, the reaction was
cooled to room temperature and the reaction was concentrated to
yield N-(3-chlorobenzyl)-2,2-diethoxyacetimidamidea (2.3 grams, 8.5
mmol, 71%) as a yellow oil.
[0933] 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.14-7.50 (m, 4H)
4.80 (br. s., 1H) 4.29 (br. s., 2H) 4.05 (q, J=7.11 Hz, 1H)
3.40-3.73 (m, 4H) 2.01 (s, 1H) 1.05-1.29 (m, 6H) LC/MS RT=1.12
mins, [M+H]=271.09
Step B: 5- and 7-Chloro-isoquinoline-3-amine
##STR01026##
[0935] In a flask was placed
N-(3-chlorobenzyl)-2,2-diethoxyacetimidamide (1.9 g, 7.02 mmol) and
sulfuric acid (14 mL, 263 mmol). This was stirred at room
temperature for 24 hours. The reaction was then neutralized to pH 9
using 10 N NaOH. The resulting precipitate was dissolved in
dichloromethane and the water layer was washed with
dichloromethane. The organic layers were combined and concentrated
to give the above regioisomers (1.1 grams, 6.1 mmol, 88%) in a 2:1
ratio of 7-chloroisoquinolin-3-amine to
5-chloroisoquinolin-3-amine. This regioisomer mixture is carried on
without further separation. 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 8.89 (s, 1H) 8.81 (s, 2H) 7.90 (d, J=2.26 Hz, 2H) 7.82 (d,
J=8.28 Hz, 1H) 7.65 (dd, J=7.28, 1.00 Hz, 1H) 7.58 (d, J=9.03 Hz,
2H) 7.44 (dd, J=9.03, 2.26 Hz, 2H) 7.07-7.19 (m, 1H) 6.83 (s, 1H)
6.64 (s, 2H) 6.28 (s, 2H) 6.05 (s, 4H) LC/MS R.T=0.955 mins,
[M+H]=181.03
Step C: 5- and 7-Chloro-3-isothiocyanato-isoquinoline
##STR01027##
[0937] In a vial was placed
7-chloroisoquinolin-3-amine/5-chloroisoquinolin-3-amine (550 mg,
3.08 mmol) and 1,1'-thiocarbonyldipyridin-2(1H)-one (858 mg, 3.70
mmol) in dichloromethane (10 mL). The reaction was stirred at room
temperature. After 18 hours, the reaction was purified on a silica
gel cartridge, eluting in 50% ethyl acetate in hexanes to give a
mixture of 7-chloro-3-isothiocyanatoisoquinoline and
5-chloro-3-isothiocyanatoisoquinoline (400 mg, 1.8 mmol, 58%) which
was used immediately in the next reaction. LC/MS RT=3.808 mins
[M+H]=221.0
Step D:
(R)--N-(5-chloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine and
(R)--N-(7-chloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2-
.2]octan]-2-amine
##STR01028##
[0939] In a vial was placed
7-chloro-3-isothiocyanatoisoquinoline/5-chloro-3-isothiocyanatoisquinolin-
e (400 mg, 1.813 mmol) and (S)-3-(aminomethyl)quinuclidin-3-ol (340
mg, 2.175 mmol) in DMF (8 mL). To this was added cesium carbonate
(1417 mg, 4.35 mmol). The reaction was allowed to stir at 50 C for
1 hour and then N,N'-diisopropylcarbodiimide (0.847 mL, 5.44 mmol)
was added to the reaction and the reaction was allowed to stir
overnight. After 18 hours the reaction was poured into water and
chloroform. The organic was collected, concentrated, and purified
on the Biotage eluting in 5-40% (10% NH4OH/Methanol) in chloroform.
The product regioisomers were collected and separated by
preparative SFC to give
(R)--N-(5-chloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2-
.2]octan]-2-amine (12.5 mg) as a light yellow solid. 1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 9.14 (s, 1H) 8.67 (br. s., 1H) 8.03
(d, J=8.28 Hz, 1H) 7.82 (d, J=7.28 Hz, 1H) 7.41 (t, J=8.03 Hz, 1H)
3.90 (br. s., 1H) 3.63 (br. s., 1H) 2.91-3.16 (m, 2H) 2.65-2.92 (m,
4H) 1.85-2.19 (m, 2H) 1.16-1.81 (m, 4H) LC/MS RT=1.578 mins
[M+H]=343.08, and
(R)--N-(7-chloroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2-
.2]octan]-2-amine (54.9 mg) was isolated as a light yellow solid.
1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.06 (s, 1H) 8.43-8.93
(m, 1 H) 8.13 (br. s., 1H) 7.82 (br. s., 1H) 7.61 (d, J=8.28 Hz,
1H) 3.89 (br. s., 1H) 3.61 (br. s., 1H) 3.31 (br. s., 1H) 2.90-3.18
(m, 2H) 2.62-2.95 (m, 4H) 1.82-2.13 (m, 2H) 1.34-1.80 (m, 3H) LC/MS
R.T=0.898 mins [M+H]=343.32.
Example 339
(R)--N-(6-Chlorothiazolo[4,5-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine
##STR01029##
[0940] Step A:
N-(3,5-Dichloropyridin-2-ylcarbamothioyl)benzamide
##STR01030##
[0942] To 3,5-dichloropyridin-2-amine (3.87 g, 23.73 mmol) in
acetone (20 mL) was added benzoyl isothiocyanate (3.2 mL, 23.73
mmol). The mixture was stirred at ambient temperature for 1 h, and
the resultant precipitate was collected by filtration (3.89 g). The
filtrate was returned to the reaction flask and heated to reflux 1
h. At this time, the reaction had gone dry. The solids were
suspended in acetone and collected by filtration to give 2.24 g. of
N-(3,5-dichloropyridin-2-ylcarbamothioyl)benzamide (5.93 g, 18.18
mmol, 77% yield). .sup.1H NMR (500 MHz, CHLOROFORM-d) .delta. ppm
12.78 (br. s., 1H) 9.25 (br. s., 1H) 8.46 (br. s., 1H) 7.94 (d,
J=7.63 Hz, 2H) 7.88 (d, J=2.44 Hz, 1H) 7.69 (t, J=7.48 Hz, 1H) 7.58
(t, J=7.78 Hz, 2H).
Step B: N-(6-Chlorothiazolo[4,5-b]pyridin-2-yl)benzamide
##STR01031##
[0944] To N-(3,5-dichloropyridin-2-ylcarbamothioyl)benzamide (5.2
g, 16 mmol) in NMP (40 mL) was added sodium methoxide (1.73 g, 32.0
mmol). The mixture was heated in a 120.degree. C. oil bath for 4 h,
cooled to ambient temperature and poured into water. The solids
were collected by filtration and purified on by silica gel
chromatography (0-5% (9:1 MeOH:NH.sub.4OH)/chloroform) to give
N-(6-chlorothiazolo[4,5-b]pyridin-2-yl)benzamide (364 mg, 1.256
mmol, 7.85% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
13.16 (br. s., 1H) 8.48-8.58 (m, 2H) 8.13-8.23 (m, 2H) 7.61-7.69
(m, 1H) 7.56 (t, J=7.53 Hz, 2H). MS (LC/MS) R.T.=2.8;
[M+H].sup.+=290.1.
Step C: 6-Chlorothiazolo[4,5-b]pyridin-2-amine
##STR01032##
[0946] N-(6-Chlorothiazolo[4,5-b]pyridin-2-yl)benzamide (364 mg,
1.256 mmol) in H.sub.2SO.sub.4 (4 ml, 75 mmol) was heated to
120.degree. C. for 30 minutes, cooled to ambient temperature,
basified with 10N NaOH and the resultant solids were collected by
filtration to afford 6-chlorothiazolo[4,5-b]pyridin-2-amine (148
mg, 0.797 mmol, 63.5% yield) .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 8.19-8.24 (m, 2H) 8.09 (br. s., 2H).
Step D: Dimethyl
6-chlorothiazolo[4,5-b]pyridin-2-ylcarbonimidodithioate
##STR01033##
[0948] 6-Chlorothiazolo[4,5-b]pyridin-2-amine (148 mg, 0.797 mmol)
was suspended in DMF (1 ml) and 16 N NaOH (0.100 ml, 1.595 mmol)
was added. The mixture was stirred 10 min at ambient temperature,
at which time, CS.sub.2 (0.120 ml, 1.993 mmol) was added. The
mixture was stirred for a further 10 min at ambient temperature and
16 N NaOH (0.100 ml, 1.595 mmol) was added. The mixture was stirred
10 min at ambient temperature, at which time, iodomethane (0.120
ml, 1.913 mmol) was added. After 15 minutes, the mixture was
diluted with water and the resultant solids were collected by
filtration. The filtrate was extracted thrice with chloroform and
dried over sodium sulfate. The crude mixture was purified by silica
gel chromatography (2-50% EtOAc/CHCl.sub.3) to afford dimethyl
6-chlorothiazolo[4,5-b]pyridin-2-ylcarbonimidodithioate (50 mg,
0.173 mmol, 21.64% yield). .sup.1H NMR (500 MHz, CHLOROFORM-d)
.delta. ppm 8.57 (s, 1H) 8.09 (s, 1H) 2.66 (s, 6H).
Step E:
(R)--N-(6-Chlorothiazolo[4,5-b]pyridin-2-yl)-4H-1'-azaspiro[oxazol-
e-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01034##
[0950] Dimethyl
6-chlorothiazolo[4,5-b]pyridin-2-ylcarbonimidodithioate (50 mg,
0.17 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol, 2HCl (40 mg, 0.17
mmol) and cesium carbonate (112 mg, 0.35 mmol) were suspended in
DMF (1 mL) and heated in an open flask on a 100.degree. C. oil
bath. After 2.5 h, the mixture was cooled to ambient temperature
and poured into water. The mixture was extracted with chloroform
4.times., dried over sodium sulfate, filtered, and concentrated to
residue. The crude residue was purified by silica gel
chromatography 5-40% (9:1 MeOH/NH.sub.4OH)/CHCl.sub.3 to afford
(R)--N-(6-chlorothiazolo[4,5-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine (20 mg, 0.056 mmol, 32.5% yield) as a
white powder. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 8.31
(d, J=2.51 Hz, 1H) 7.93 (d, J=2.26 Hz, 1H) 3.99 (d, J=10.04 Hz, 1H)
3.67 (d, J=10.04 Hz, 1H) 3.20-3.39 (m, 2H) 2.86-3.05 (m, 3H)
2.67-2.87 (m, 2H) 2.03-2.19 (m, 2H) 1.74 (ddd, J=13.87, 9.35, 4.64
Hz, 1H) 1.44-1.65 (m, 2H). MS (LC/MS) R.T.=1.49;
[M+H].sup.+=350.2.
Example 340
(R)--N-(5-fluorothiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine
##STR01035##
[0951] Step A: 5-Fluorothiazolo[5,4-b]pyridin-2-amine
##STR01036##
[0953] 6-Fluoropyridin-3-amine (4 g, 35.7 mmol) was added to a
3-neck flask containing a mechanically stirred suspension of
potassium rhodanate (27.7 g, 285 mmol) in acetic acid (89 mL) at
0.degree. C. The flask was then fitted with an addition funnel
charged with bromine (5.70 mL, 111 mmol) in acetic acid (29.7 mL).
The bromine solution was added over 30 min and the solution turned
into a viscous yellow mixture. After bromine addition was complete,
the reaction mixture was allowed to warm to ambient temperature and
stirred for 16 h. Water (30 mL) was added and the mixture was
heated to 85.degree. C. for 20 min before the solids were filtered
and washed with water and methanol to give
5-fluorothiazolo[5,4-b]pyridin-2-amine (4.18 g, 24.71 mmol, 69.2%
yield) as a yellow solid.
Step B: Dimethyl
5-fluorothiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate
##STR01037##
[0955] 5-Fluorothiazolo[5,4-b]pyridin-2-amine (2.0 g, 11.8 mmol)
was reacted according to the method of Example 339, STEP D to
provide dimethyl
5-fluorothiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate (2.17 g,
67% yield) as a yellow solid. .sup.1H NMR (500 MHz, chloroform-d)
.delta. ppm 8.14 (dd, J=8.55, 7.02 Hz, 1 H) 6.98 (dd, J=8.70, 1.98
Hz, 1H) 2.63 (s, 6H).
Step C:
(R)--N-(5-Fluorothiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazol-
e-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01038##
[0957] Dimethyl
5-fluorothiazolo[5,4-b]pyridin-2-ylcarbonimidodithioate (100 mg,
0.37 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol, 2HCl (84 mg, 0.37
mmol) and cesium carbonate (238 mg, 0.73 mmol) were suspended in
DMF (2 mL) and heated in an open flask in a 100.degree. C. oil
bath. After 2.5 h, the mixture was cooled to ambient temperature
and poured into water. The mixture was extracted with chloroform
4.times., dried over sodium sulfate, filtered, and concentrated.
The crude residue was purified by silica gel chromatography 5-40%
(9:1 MeOH/NH.sub.4OH)/CHCl.sub.3 to afford
((R)--N-(5-fluorothiazolo[5,4-b]pyridin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octan]-2-amine (85 mg, 66% yield) as a white powder.
.sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 9.17 (br. s., 1H)
7.89 (dd, J=8.66, 6.90 Hz, 1H) 6.91 (dd, J=8.53, 2.01 Hz, 1H) 4.04
(d, J=9.54 Hz, 1H) 3.73 (d, J=9.54 Hz, 1H) 3.43 (dd, J=14.93, 1.63
Hz, 1H) 3.08 (dd, J=15.06, 1.76 Hz, 1H) 2.84-3.05 (m, 4 H)
2.16-2.28 (m, 2H) 1.75-1.87 (m, 1H) 1.53-1.70 (m, 2H). MS (LC/MS)
R.T.=1.37; [M+H].sup.+=334.2.
Example 341
(R)--N-(6-methylthiazolo[5,4-b]pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine
##STR01039##
[0958] Step A: 3-Bromo-5-methylpyrazin-2-amine
##STR01040##
[0960] To a suspension of 5-methylpyrazin-2-amine (1.09 g, 10 mmol)
in chloroform (100 mL) was added pyridine (0.85 mL, 10.5 mmol). The
mixture was stirred in a foil-wrapped flask fitted with an addition
funnel, and a solution of bromine (0.54 mL, 10.5 mmol) in
chloroform (10 mL) was added dropwise over 10 min. The mixture was
allowed to react an additional 20 minutes after addition was
complete and then poured into a separatory funnel containing 10 mL
water. The phases were separated and the organics washed again with
water, dried over sodium sulfate, filtered and concentrated in
vacuo. The resulting red oil was purified by silica gel
chromatography with 12-100% EtOAc/hexanes. The major UV active peak
was collected to give 3-bromo-5-methylpyrazin-2-amine (1.06 g, 5.64
mmol, 56.4% yield) as a cream-colored solid. .sup.1H NMR (500 MHz,
DMSO-d6) .delta. ppm 7.85 (s, 1H) 6.42 (br. s., 2H) 2.25 (s, 3H).
MS (LC/MS) R.T.=0.93; [M+H].sup.+=189.9.
Step B: Ethyl 6-methylthiazolo[5,4-b]pyrazin-2-ylcarbamate
##STR01041##
[0962] To 3-bromo-5-methylpyrazin-2-amine (850 mg, 4.52 mmol) was
added ethoxycarbonyl isothiocyanate (0.51 mL, 4.52 mmol) followed
by toluene (1 mL). The mixture was placed in a preheated
100.degree. C. oil bath, and within a minute, all the solids had
dissolved. After 15 min, all had seized to a solid mass. After an
additional 15 minutes, methanol (.about.5 mL) was added and the
mixture refluxed to digest the solids. The mixture was cooled to rt
after 5 minutes and the solids collected by filtration, washing
with methanol to give ethyl
6-methylthiazolo[5,4-b]pyrazin-2-ylcarbamate (680 mg, 2.85 mmol,
63.1% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta. ppm 12.48 (br.
s., 1H) 8.48 (s, 1H) 4.30 (q, J=7.03 Hz, 2H) 2.56-2.65 (m, 3H) 1.31
(t, J=7.15 Hz, 3H). MS (LC/MS) R.T.=2.10; [M+H].sup.+=239.17.
Step C: 6-Methylthiazolo[5,4-b]pyrazin-2-amine
##STR01042##
[0964] Ethyl 6-methylthiazolo[5,4-b]pyrazin-2-ylcarbamate (660 mg,
2.77 mmol) was suspended in 1N NaOH (15 ml, 15.00 mmol) and heated
in a 100.degree. C. oil bath for 4 h and cooled to ambient
temperature. The mixture was acidified with 1N HCl and the
resulting precipitate was filtered off and washed with ether,
affording 6-methylthiazolo[5,4-b]pyrazin-2-amine (233 mg, 1.402
mmol, 50.6% yield) as a yellow powder. .sup.1H NMR (500 MHz,
DMSO-d6) .delta. ppm 8.26 (s, 2H) 8.12 (s, 1H) 2.43 (s, 3H). MS
(LC/MS) R.T.=0.66; [M+H].sup.+=167.0.
Step D: Dimethyl
6-methylthiazolo[5,4-b]pyrazin-2-ylcarbonimidodithioate
##STR01043##
[0966] 6-Methylthiazolo[5,4-b]pyrazin-2-amine (0.22 g, 1.32 mmol)
was reacted according to the method of Example 339, STEP D to
provide dimethyl
6-methylthiazolo[5,4-b]pyrazin-2-ylcarbonimidodithioate (265 mg,
59% yield) as a yellow solid. .sup.1H NMR (500 MHz, CHLOROFORM-d)
.delta. ppm 8.45 (s, 1H) 2.66-2.68 (m, 9H).
Step E:
(R)--N-(6-Methylthiazolo[5,4-b]pyrazin-2-yl)-4H-1'-azaspiro[oxazol-
e-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01044##
[0968] Dimethyl
6-methylthiazolo[5,4-b]pyrazin-2-ylcarbonimidodithioate (150 mg,
0.56 mmol), (S)-3-(aminomethyl)quinuclidin-3-ol, 2HCl (140 mg, 0.61
mmol) and cesium carbonate (450 mg, 1.39 mmol) were suspended in
DMF (3 mL) and heated in an open flask in a 100.degree. C. oil
bath. After 2.5 h, the mixture was cooled to ambient temperature
and poured into water. The mixture was extracted with chloroform
4.times., dried over sodium sulfate, filtered, and concentrated to
residue. The crude residue was purified by silica gel
chromatography 5-40% (9:1 MeOH/NH.sub.4OH)/CHCl.sub.3 to afford
(R)--N-(6-methylthiazolo[5,4-b]pyrazin-2-yl)-4H-1'-azaspiro[oxazole-5,3'--
bicyclo[2.2.2]octan]-2-amine (51 mg, 27% yield) as a white powder.
.sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 9.45 (br. s., 1H)
8.26 (s, 1H) 4.02 (d, J=9.79 Hz, 1 H) 3.69 (d, J=9.79 Hz, 1H) 3.39
(dd, J=14.93, 1.63 Hz, 1H) 2.72-3.06 (m, 5H) 2.60 (s, 3H) 2.11-2.22
(m, 2H) 1.70-1.82 (m, 1H) 1.49-1.64 (m, 2H). MS (LC/MS) R.T.=1.15;
[M+H].sup.+=331.2.
Example 342
(R)--N-(6,7-dimethoxyisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo-
[2.2.2]octan]-2-amine
##STR01045##
[0969] Step A:
N-(3,4-Dimethoxybenzyl)-2,2-diethoxyacetimidamide
##STR01046##
[0971] A mixture of methyl 2,2-diethoxyacetimidate (294 mg, 1.824
mmol), (3,4-dimethoxyphenyl)methanamine (312 mg, 1.866 mmol), and
MeOH (1.5 mL) was heated with stirring in an oil bath at 70.degree.
C. for 2 hrs and allowed to cool to room temperature. The volatile
components were removed in vacuo. The residue was carried on
without further purification. LCMS RT 0.96 min, MH.sup.+=297.2, 1H
NMR (400 MHz, CHLOROFORM-d) .delta. ppm 6.79-6.93 (3H, m), 4.94
(1H, s), 4.41 (2H, s), 3.84-3.90 (6H, m), 3.52-3.70 (4H, m),
1.19-1.28 (6H, m).
Step B: 6,7-Dimethoxyisoquinolin-3-amine
##STR01047##
[0973] To N-(3,4-dimethoxybenzyl)-2,2-diethoxyacetimidamide (541
mg, 1.825 mmol) was added sulfuric acid (0.95 mL, 1.825 mmol) was
added. After cooling, the reaction was allowed to stand at ambient
temperature overnight. It was added dropwise to ice, then the
resulting solution was neutralized with conc. NaOH, and the aqueous
phase was twice extracted with ethyl acetate. The combined ethyl
acetate fractions were dried over magnesium sulfate. The drying
agent was filtered off and the solvent evaporated. The residue was
purified by column chromatography in 5% methanol/ethyl acetate,
collecting the main component. Yield 105.7 mg brown solid
(28%).
[0974] LCMS RT 0.72 min, MH.sup.+=205.1; 1H NMR (500 MHz,
CHLOROFORM-d) .delta. ppm 8.59 (1H, s), 7.03 (1H, s), 6.80 (1H, s),
6.70 (1H, s), 4.00 (3H, s), 3.98 (3H, s)
Step C: 3-Isothiocyanato-6,7-dimethoxyisoquinoline
##STR01048##
[0976] To a stirring solution of
1,1'-thiocarbonyldipyridin-2(1H)-one (124.6 mg, 0.536 mmol) in
dichloromethane (3 mL) was added a suspension of
6,7-dimethoxyisoquinolin-3-amine (105 mg, 0.514 mmol) in
dichloromethane (5 mL). The reaction mixture was applied directly
to a Biotage column in 15-25% ethyl acetate/hexane followed by 100%
ethyl acetate to give 3-isothiocyanato-6,7-dimethoxyisoquinoline as
a white solid. Yield 69.4 mg (55%) NMR 1H NMR (500 MHz,
CHLOROFORM-d) .delta. ppm 8.81 (1H, s), 7.31 (1H, s), 7.16 (1H, s),
6.98 (1H, s), 3.99 (6H, s)
Step D:
(R)--N-(6,7-dimethoxyisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-
-bicyclo[2.2.2]octan]-2-amine
##STR01049##
[0978] To a stirring suspension of
(S)-3-(ammoniomethyl)-3-hydroxy-1-azoniabicyclo[2.2.2]octane
chloride (73 mg, 0.319 mmol) and cesium carbonate (221 mg, 0.678
mmol) in DMF (5 mL) was added a solution of
3-isothiocyanato-6,7-dimethoxyisoquinoline (69 mg, 0.280 mmol) in
DMF (1.5 mL) and the reaction mixture was allowed to stir at room
temperature for 3 days. To the solution was then added a solution
of N,N'-methanediylidenedipropan-2-amine (69 mg, 0.547 mmol) in DMF
(0.4 mL) and the reaction was allowed to stand at room temperature
for 14 days. Another 69 mg di-isopropylcarbodiimide was added in
.about.0.4 mL acetonitrile and the reaction was allowed to stand
for 1 more day, then it was evaporated in vacuo and the residue
subjected to preparative HPLC to yield
(R)--N-(6,7-dimethoxyisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicycl-
o[2.2.2]octan]-2-amine 20.9 mg. (8%) LCMS RT 0.98 min,
MH.sup.+=403.2 1H NMR (500 MHz, CHLOROFORM-d) .delta. ppm 8.73 (1H,
s), 7.28 (1H, s), 7.09 (1H, s), 6.96 (1H, s), 4.01 (3H, s), 4.00
(3H, s), 3.96 (1H, d, J=8.5 Hz), 3.63 (1H, d, J=9.2 Hz), 3.39 (1 H,
d, J=15.0 Hz), 2.79-3.10 (5H, m), 2.24 (1H, br. s.), 2.15 (1H, br.
s.), 1.75 (1H, dddd, J=13.9, 9.3, 4.7, 4.4 Hz), 1.59-1.68 (1H, m),
1.48-1.59 (1H, m)
Example 343
(R)--N-(5,6,7,8-Tetrahydroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine,
##STR01050##
[0979] Step A: 5,6,7,8-Tetrahydroisoquinolin-3-amine
##STR01051##
[0981] A mixture of isoquinolin-3-amine (239 mg, 1.66 mmol),
platinum(IV)oxide (28 mg, 0.123 mmol), and TFA (6 mL) was
hydrogenated in the Parr apparatus for 3 hrs. The reaction mixture
was filtered with the aid of ethyl acetate. The filtrate was
evaporated in vacuo and the residue was partitioned between 10%
aqueous sodium carbonate and ethyl acetate. The layers were
separated, the aqueous phase was washed again with ethyl acetate,
and the combined organic layers were washed with brine and dried
over magnesium sulfate. The drying agent was filtered off and the
solvent evaporated. The material was purified by column
chromatography in ethyl acetate to give 139.8 mg (57%)
5,6,7,8-tetrahydroisoquinolin-3-amine as a yellow-white solid. LCMS
RT 0.75 min, MH.sup.+=149.1 1H NMR (400 MHz, CHLOROFORM-d) .delta.
ppm 7.78 (1H, s), 6.26 (1H, s), 4.29 (2H, br. s.), 2.64 (4H, ddd,
J=10.6, 5.8, 5.5 Hz), 1.64-1.87 (4H, m).
Step B: 3-Isothiocyanato-5,6,7,8-tetrahydroisoquinoline
##STR01052##
[0983] To a stirring solution of
1,1'-thiocarbonyldipyridin-2(1H)-one (222 mg, 0.956 mmol) in
dichloromethane (5 mL) was added a solution of
5,6,7,8-tetrahydroisoquinolin-3-amine (139 mg, 0.938 mmol) in
dichloromethane (1.5 mL) and the resulting solution was stirred at
room temperature overnight. e reaction mixture was applied directly
to a Biotage column in 20-25% ethyl acetate/hexane, collecting the
first major peak. Yield 141.8 mg (79%)
3-iothiocyanato-5,6,7,8-tetrahydroisoquinoline. NMR 1H NMR (500
MHz, CHLOROFORM-d) .delta. ppm 8.10 (1H, s), 6.84 (1H, s), 2.74
(4H, t, J=6.3 Hz), 1.81 (4H, dd, J=4.0, 2.7 Hz).
Step C:
(S)-1-((3-Hydroxyquinuclidin-3-yl)methyl)-3-(5,6,7,8-tetrahydroiso-
quinolin-3-yl)thio-urea
##STR01053##
[0985] To a stirring suspension of
(S)-3-(ammoniomethyl)-3-hydroxy-1-azoniabicyclo[2.2.2]octane
chloride (183 mg, 0.799 mmol) and cesium carbonate (550 mg, 1.688
mmol) in DMF (10 mL) was added a solution of
3-isothiocyanato-5,6,7,8-tetrahydroisoquinoline (141 mg, 0.741
mmol) in DMF (2 mL) and the reaction mixture was stirred at room
temperature for 10 days. Then the solvent was removed in vacuo and
the residue was taken up in methanol and purified on a silica gel
column in 1% NH.sub.4OH/9% MeOH/90% CHCl.sub.3 followed by
purification via preparative HPLC. The isolated product was
partitioned between aqueous sodium carbonate and ethyl acetate. The
aqueous phase was washed again with ethyl acetate and the combined
organic phases were washed with brine, dried over magnesium
sulfate, filtered and the solvent evaporated. The residue was
re-purified on a silica gel column in 1% NH.sub.4OH/9% MeOH/90%
CHCl.sub.3 to give
(S)-1-((3-hydroxyquinuclidin-3-yl)methyl)-3-(5,6,7,8-tetrahydroisoquinoli-
n-3-yl)thio-urea, yield 98 mg (35%). 1H NMR (500 MHz, CHLOROFORM-d)
.delta. ppm 12.02 (1H, br s), 8.23 (1H, br s), 7.88 (1H, s), 6.41
(1H, s), 4.08 (1H, dd), 3.89 (1H, dd), 3.46 (2H, s), 3.05-2.80 (5H,
m), 2.69 (4H, dt), 2.10 (1H, br s), 1.93 (1H, br s), 1.85 (1H, m),
1.77 (4H, m), 1.66 (1H, m), 1.45 (1H, m); LCMS RT 1.11 min,
MH.sup.+=447.1.
Step D:
(R)--N-(5,6,7,8-Tetrahydroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-
-5,3'-bicyclo-[2.2.2]octan]-2-amine
##STR01054##
[0987] To a solution of
(S)-1-((3-hydroxyquinuclidin-3-yl)methyl)-3-(5,6,7,8-tetrahydroisoquinoli-
n-3-yl)thiourea (98 mg, 0.283 mmol) in DMF (2 mL) was added a
solution of N,N'-methanediylidenedipropan-2-amine (41 mg, 0.325
mmol) in DMF (0.4 mL) and the resulting solution was allowed to
stand at room temperature for 7 days.
[0988] Another 53 mg di-isopropylcarbodiimide was added in
.about.0.4 mL acetonitrile and the reaction was allowed to stand
for 1 more day. The reaction mixture was then evaporated in vacuo
and subjected to preparative HPLC followed by silica gel
chromatography in 1% NH.sub.4OH/9% MeOH/90% CHCl.sub.3 to give
(R)--N-(5,6,7,8-tetrahydroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo-[2.2.2]octan]-2-amine, yield 6.4 mg. (7%)
[0989] LCMS RT 0.80 min, MH.sup.+=313.2; 1H NMR (500 MHz,
CHLOROFORM-d) .delta. ppm 7.91 (1H, s), 6.75-6.95 (1H, m), 3.91
(1H, d, J=9.5 Hz), 3.60 (1H, d, J=9.5 Hz), 3.38 (1H, s), 2.93-3.13
(4H, m), 2.86-2.93 (2H, m), 2.69 (4H, td, J=11.4, 6.1 Hz),
2.21-2.32 (1H, m), 2.16 (1H, br. s.), 1.72-1.85 (5H, m), 1.64 (1H,
dd, J=7.2, 4.4 Hz), 1.56 (1H, dt, J=7.0, 2.6 Hz)
Example 344
(R)--N-(6-Chloro-7-methoxyisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine
##STR01055##
[0990] Step A:
N-(4-Chloro-3-methoxybenzyl)-2,2-diethoxyacetimidamide
##STR01056##
[0992] To a mixture of (4-chloro-3-methoxyphenyl)methanamine (392
mg, 2.284 mmol) (as the hydrochloride salt), Hunig's Base (0.5 mL,
2.86 mmol), and MeOH (2 mL) was added a solution of methyl
2,2-diethoxyacetimidate (295 mg, 1.83 mmol) in MeOH (1 mL) and the
resulting mixture was heated with stirring in an oil bath at
70.degree. C. for 2 hrs and allowed to cool to room temperature
over 3 days. The solvent was removed in vacuo and the residue was
used without further purification.
Step B: 6-Chloro-7-methoxyisoquinolin-3-amine
##STR01057##
[0994] To cold (0.degree. C.) sulfuric acid (0.071 mL, 1.330 mmol)
was added gradually a suspension of
N-(4-chloro-3-methoxybenzyl)-2,2-diethoxyacetimidamide (400 mg,
1.330 mmol) in dichloromethane (5 mL) and the resulting mixture was
allowed to warm up to room temperature overnight. The reaction
mixture was then added slowly to crushed ice, then the mixture was
made strongly basic by adding conc. NaOH solution and the organic
components were extracted into ethyl acetate. The organic fraction
was dried over magnesium sulfate, filtered, and the solvent was
evaporated. The residue was subjected to silica gel chromatography
in ethyl acetate to give 110.8 mg. (40%)
6-chloro-7-methoxyisoquinolin-3-amine LCMS RT 0.95 min,
MH.sup.+=209.0, 211.0; 1H NMR (500 MHz, CHLOROFORM-d) .delta. ppm
8.66 (1H, s), 7.78 (1 H, s), 6.83 (1H, s), 6.63 (1H, s), 4.36-4.58
(2H, m), 3.99 (3H, s).
Step C: 6-Chloro-3-isothiocyanato-7-methoxyisoquinoline
##STR01058##
[0996] To a stirring solution of
1,1'-thiocarbonyldipyridin-2(1H)-one (142 mg, 0.611 mmol) in
dichloromethane (5 mL) was added a suspension of
6-chloro-7-methoxyisoquinolin-3-amine (110 mg, 0.527 mmol) in
dichloromethane (5.00 mL) and the reaction mixture was stirred at
room temperature overnight. The reaction mixture was applied
directly to a silica gel column in 10% ethyl acetate/hexane to
yield 106 mg (80%) 6-Chloro-3-isothiocyanato-7-methoxyisoquinoline.
1H NMR (500 MHz, CHLOROFORM-d) .delta. ppm 8.90 (1H, s), 7.99 (1H,
s), 7.36 (1H, s), 7.08 (1H, s), 4.05 (3H, s).
Step D:
(S)-1-(6-Chloro-7-methoxyisoquinolin-3-yl)-3-((3-hydroxyquinuclidi-
n-3-yl)methyl)thiourea
##STR01059##
[0998] To a stirring suspension of
(S)-3-(ammoniomethyl)-3-hydroxy-1-azoniabicyclo[2.2.2]octane
chloride (111 mg, 0.484 mmol) in DMF (3 mL) was added Hunig's Base
(0.5 mL, 2.86 mmol) and the resulting mixture was stirred at room
temperature for 35 minutes. Then was added a suspension of
6-chloro-3-isothiocyanato-7-methoxyisoquinoline (106 mg, 0.423
mmol) in DMF (3 mL) and the reaction mixture was allowed to stir at
room temperature overnight. The solvent was evaporated in vacuo and
the residue subjected to a silica gel column in 1% NH.sub.4OH/9%
MeOH/90% CHCl.sub.3 to give 140.3 mg (82%) as a white solid,
(S)-1-(6-chloro-7-methoxyisoquinolin-3-yl)-3-((3-hydroxyquinuclidin-3-yl)-
methyl)thiourea. LCMS RT 1.28 min, MH.sup.+=407.1, 409.0, 1H NMR
(500 MHz, CHLOROFORM-d) .delta. ppm 11.85 (1H, t, J=5.2 Hz), 9.15
(1H, br. s.), 8.63 (1H, s), 7.76 (1H, s), 6.92 (1H, s), 6.89 (1H,
s), 4.14 (1H, dd, J=13.9, 5.6 Hz), 3.99 (2H, s), 3.90 (1H, dd,
J=14.0, 5.2 Hz), 3.46 (3H, s), 2.89-3.01 (2H, m), 2.75-2.88 (3H,
m), 2.03-2.15 (1H, m), 1.89-1.96 (1H, m), 1.77-1.88 (1H, m), 1.63
(1H, ddd, J=13.4, 6.1, 3.4 Hz), 1.32-1.45 (1H, m).
Step E:
(R)--N-(6-Chloro-7-methoxyisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-
-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01060##
[1000] To a solution of
(S)-1-(6-chloro-7-methoxyisoquinolin-3-yl)-3-((3-hydroxyquinuclidin-3-yl)-
methyl)thiourea (140 mg, 0.344 mmol) in DMF (2.5 mL) was added a
solution of N,N'-methanediylidenedipropan-2-amine (101 mg, 0.800
mmol) in DMF (0.5 mL) and the resulting mixture was allowed to
stand at room temperature for 10 days. The solvent was then
evaporated in vacuo. The residue was dissolved in .about.5 mL MeOH
(some white solid was filtered off, 09) and subjected to
preparative HPLC purification followed by silica gel chromatography
in 1% NH.sub.4OH/9% MeOH/90% CHCl.sub.3 to give 31 mg (24%)
(R)--N-(6-chloro-7-methoxyisoquinolin-3-yl)-4H-1'-azaspiro[oxazole--
5,3'-bicyclo[2.2.2]octan]-2-amine LCMS RT 1.04 min, MH.sup.+=373.2,
375.2 1H NMR (500 MHz, MeOD) .delta. ppm 8.78-8.89 (1H, m),
7.90-8.01 (1H, m), 7.14-7.23 (1H, m), 4.01 (3H, s), 3.95-3.99 (1H,
m), 3.62-3.71 (1H, m), 3.19-3.25 (1H, m), 3.06-3.13 (1H, m),
2.90-3.00 (2H, m), 2.76-2.89 (2H, m), 2.10-2.21 (2H, m), 1.69-1.83
(2H, m), 1.56-1.68 (1H, m).
Example 345
(R)--N-(5-fluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.-
2]octan]-2-amine
##STR01061##
[1001] Step A: 7-Fluoroisoquinolin-3-amine and
5-fluoroisoquinolin-3-amine
##STR01062##
[1003] A mixture of methyl 2,2-diethoxyacetimidate (1.5 g, 10.2
mmol) and (3-fluorophenyl)methanamine (1.275 g, 10.2 mmol) in
methanol (10 ml) was stirred for 2 h at 70 degree. The solvent was
evaporated to an oil, 2,2-diethoxy-N-(3-fluorobenzyl)acetimidamide
(2.5 g. 100%). .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm
7.3-6.9 (m, 4H), 4.9 (s, 1H), 4.4 (s, 2H), 3.8-3.5 (m, 4H),
1.27-1.20 (q, 6H). MS (LCMS in ammonium acetate system)
[M+H]=254.7.
[1004] To the above prepared oil (2.1 g) in CH.sub.2Cl.sub.2 (3 ml)
at RT was added 2 ml concentrated sulfuric acid very slowly and
then it was stirred at RT overnight. The mixture was poured into
ice water and neutralized to pH=8 with NaOH (10N). The mixture was
extracted with EtOAc (100 ml.times.3). The organic layers were
combined and washed with water, brine and dried over
Na.sub.2SO.sub.4 to obtain a crude solid, 1.05 g which was used
directly without further purification.
Step B: 5-Fluoro-3-isothiocyanatoisoquinoline
##STR01063##
[1006] To a solution of the above crude mixture (1050 mg, 6.47
mmol) in methylene chloride (30 ml) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (1500 mg, 6.5 mmol). The pink
solution was stirred for 2 h at RT and then the solvent was
evaporated to obtain a crude red solid which was purified on the
Biotage (0-25% ethyl acetate-hexane), isolating the first peak,
identified as 7-fluoro-3-isothiocyanatoisoquinoline (635 mg, 3.11
mmol, 48%). .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 9.08 (s,
1H), 7.9-7.8 (m, 1H), 7.7-7.6 (m, 1H), 7.6-7.5 (m, 2H), and the
second isomer, 5-fluoro-3-isothiocyanatoisoquinoline (27.5 mg,
0.135 mmol, 2.1%). .sup.1H NMR (500 MHz, CDCl3) .delta. ppm 9.15
(s, 1H), 7.9-7.8 (m, 1H), 7.7 (s, 1H), 7.6-7.5 (m, 1H), 7.5-7.4 (m,
1H).
Step C:
(R)--N-(5-Fluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR01064##
[1008] To a solution of 5-fluoro-3-isothiocyanatoisoquinoline (27.5
mg, 0.135 mmol) in DMF (5 ml) was added
(S)-3-(aminomethyl)quinuclidin-3-ol dihydrochloride (25 mg, 0.160
mmol) and cesium carbonate (110 mg, 0.337 mmol). The mixture was
stirred at 40 degree overnight. To the reaction was then added
N,N'-methanediylidenedipropan-2-amine (0.12 g, 0.95 mmol) and the
mixture was stirred at 40 degree overnight. The solvent was
evaporated and the crude solid was purified on the Biotage, first
in 100% ethyl acetate and then 0-25% 9:1 methanol:ammonium
hydroxide-chloroform in a second run to afford
(R)--N-(5-fluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2-
.2]octan]-2-amine (29.4 mg, 0.086 mmol, 64.1%). .sup.1H NMR (500
MHz, MeOD) .delta. ppm 9.09 (s, 1H), 7.80-7.75 (d, 1H), 7.50-7.30
(m, 3H), 4.02-4.00 (d, 1H), 3.70-3.60 (d, 1H), 3.4-3.2 (m, 1H),
3.2-3.1 (m, 1H), 3.1-2.9 (m, 2H), 2.9-2.8 (m, 2H), 2.2-2.1 (m, 2H),
1.8-1.6 (m, 3H). MS (LCMS) [M+H]=327.08.
Example 346
(R)--N-(8-Bromo-5-fluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicy-
clo[2.2.2]octan]-2-amine
##STR01065##
[1009] Step A: Methyl 2,2-diethoxyacetimidate
##STR01066##
[1011] To a solution of sodium methoxide (13.38 g, 248 mmol) in
MeOH (300 mL) was added dropwise 2,2-diethoxyacetonitrile (17.22
mL, 124 mmol). The resulting mixture was stirred at rt for 16 h.
The reaction was then diluted with 300 mL of water and the product
was extracted with 3.times.250 mL of Et.sub.2O. The combined
Et.sub.2O layers were washed with 100 mL of brine, dried over
sodium sulfate, and evaporated to give the desired product as a
colorless liquid (15.3 g, 95 mmol, 77% yield). .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. ppm 7.86 (1H, br. s.), 4.77 (1H, s), 3.78
(3H, s), 3.55 (4H, qd, J=6.97, 4.28 Hz), 1.21 (7H, t, J=7.05
Hz).
Step B: (2-Bromo-5-fluorophenyl)methanamine
##STR01067##
[1013] To a solution of 2-bromo-5-fluorobenzonitrile (3.0 g, 15.00
mmol) and NaBH.sub.4 (1.419 g, 37.5 mmol) in THF (30 mL) was slowly
added TFA (3.47 mL, 45.0 mmol) over a period of 20 min. The
resulting mixture was stirred at rt for 16 hours, then MeOH (10 mL)
was added and the mixture was stirred for another 30 min. It was
then diluted with EtOAc (200 mL), washed with water, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was purified on an 80
g Thompson silica cartridge (3% to 100% B in Hexanes, 1200 mL, B:
10% MeOH in EtOAc). The desired product was obtained as a colorless
oil (1.70 g, 8.33 mmol, 55.5% yield). LC/MS (0.647 min, MH+:
205.92). .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 7.59 (1H,
dd, J=8.85, 5.49 Hz), 7.43 (1H, dd, J=10.07, 3.05 Hz), 7.05 (1H,
td, J=8.47, 3.20 Hz), 3.72 (3H, s), 2.00 (2H, br. s.).
Step C: N-(2-bromo-5-fluorobenzyl)-2,2-diethoxyacetimidamide
##STR01068##
[1015] A mixture of methyl 2,2-diethoxyacetimidate (1.264 g, 7.84
mmol) and (2-bromo-5-fluorophenyl)methanamine (1.6 g, 7.84 mmol) in
MeOH (20 mL) was stirred at 75.degree. C. for 2 h. The solvent was
evaporated off to provide the crude product as a pale yellow solid.
N-(2-bromo-5-fluorobenzyl)-2,2-diethoxyacetimidamide (2.61 g, 7.83
mmol, 100% yield) which was used directly in the next step. LC/MS
(1.188 min, MH+: 334.92). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 7.44 (1H, dd, J=8.69, 5.16 Hz), 7.15 (1H, dd, J=9.44, 2.90 Hz),
6.81 (1H, td, J=8.25, 3.15 Hz), 5.48 (1H, br. s.), 4.90 (1H, s),
4.42 (2H, s), 3.49-3.67 (4H, m), 1.17-1.25 (6H, m).
Step D: 8-bromo-5-fluoroisoquinolin-3-amine
##STR01069##
[1017] To a mixture of
N-(2-bromo-5-fluorobenzyl)-2,2-diethoxyacetimidamide (1.6 g, 4.80
mmol) and dichloromethane (2 mL) was added H.sub.2SO.sub.4 (1.280
mL, 24.01 mmol). The reaction mixture was stirred at 80.degree. C.
for 16 hours, cooled to rt., diluted with EtOAc (100 mL), quenched
with ice-water, and neutralized with NaHCO.sub.3 solution. The
organic phase was washed with water, dried over Na.sub.2SO.sub.4,
and evaporated. The residue was purified on an 80 g Thompson silica
cartridge (3% to 100% EtOAc in Hexanes, 1200 mL). The desired
product was obtained as a pale yellow solid (0.47 g, 1.950 mmol,
40.6% yield).
[1018] LC/MS (1.052 min, MH+: 242.99). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 8.95 (1 H, s), 7.36 (1H, dd, J=8.06, 4.53
Hz), 7.23 (1H, dd, J=10.58, 8.06 Hz), 6.68 (1H, s), 6.52 (2H, s),
.sup.19F NMR (376 MHz, DMSO-d.sub.6) .delta. ppm -126.83 (1F,
s).
Step E: 8-bromo-5-fluoro-3-isothiocyanatoisoquinoline
##STR01070##
[1020] To a mixture of 8-bromo-5-fluoroisoquinolin-3-amine (0.30 g,
1.245 mmol) and dichloromethane (5 mL) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (0.318 g, 1.369 mmol). The
resulting mixture was stirred at rt for 3 hours. The product was
directly purified on a 40 g Thompson silica cartridge (3% to 100%
EtOAc in Hexanes, 1200 mL). The desired product was obtained as a
off-white solid (0.23 g, 0.812 mmol, 65.3% yield). LC/MS (2.098
min, MH+: 284.93). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
9.38 (1H, s), 8.04 (1H, dd, J=8.31, 4.78 Hz), 7.96 (1H, s), 7.67 (1
H, dd, J=9.82, 8.31 Hz). .sup.19F NMR (376 MHz, DMSO-d.sub.6)
.delta. ppm -122.13 (1F, s).
Step F:
(R)--N-(8-bromo-5-fluoroisoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5-
,3'-bicyclo[2.2.2]octan]-2-amine
##STR01071##
[1022] To a mixture of
8-bromo-5-fluoro-3-isothiocyanatoisoquinoline (0.22 g, 0.777 mmol)
and (S)-3-(aminomethyl)quinuclidin-3-ol, 2HCl (0.196 g, 0.855 mmol)
in DMF (5 mL) was added Cs.sub.2CO.sub.3 (0.633 g, 1.943 mmol). The
resulting mixture was stirred at 40.degree. C. for 16 hours.
N,N'-methanediylidenedipropan-2-amine (0.294 g, 2.328 mmol) was
added and the stirring was continued at 40.degree. C. for another
16 hours. The solvent was evaporated. The product was directly
purified on 40 g Thompson silica cartridge (3% to 100% B in
hexanes, 1500 mL. B: 20% MeOH in EtOAc). The product was further
recrystallized from EtOAc. The desired product was obtained as a
off-white solid (0.11 g, 0.261 mmol, 33.6% yield). LC/MS (0.997
min, MH+: 406.98). .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.30
(1H, s), 7.60 (1H, dd, J=8.18, 4.66 Hz), 7.22 (1H, dd, J=10.07,
8.06 Hz), 4.00 (1H, d, J=10.07 Hz), 3.69 (1H, d, J=10.07 Hz), 3.23
(1H, d, J=15.36 Hz), 3.07-3.13 (1H, m), 2.90-2.96 (2H, m),
2.76-2.85 (2 H, m), 2.15 (2H, br. s.), 1.69-1.81 (2H, m), 1.58-1.67
(1H, m). .sup.19F NMR (376 MHz, MeOD) .delta. ppm -126.72 (1F,
s).
Example 347
(R)--N-(6-(1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine
##STR01072##
[1023] Step A: 4-Chloro-6-(1H-imidazol-1-yl)pyrimidine
##STR01073##
[1025] A mixture of 4,6-dichloropyrimidine (5.96 g, 40 mmol),
imidazole (2.72 g, 40.0 mmol), and potassium carbonate (5.53 g,
40.0 mmol) was stirred in DMF (50 mL) at room temperature for 18 h.
The reaction was diluted into 500 mL water and extracted five times
with 150 mL EtOAc. The combined organic layers were concentrated
and purified by flash chromatography on a 240 g silica gel
cartridge with 0 to 10% methanol in ethyl acetate to yield
4-chloro-6-(1H-imidazol-1-yl)pyrimidine (4.56 g, 63% yield).
.sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 8.85 (1H, d, J=0.76
Hz), 8.45 (1H, s), 7.62 (1H, t, J=1.51 Hz), 7.34 (1H, d, J=0.76
Hz), 7.23 (1H, dd, J=1.38, 0.88 Hz). LCMS: RT=0.47 min,
MH+=181.1.
Step B: 6-(1H-Imidazol-1-yl)pyrimidin-4-amine
##STR01074##
[1027] 4-Chloro-6-(1H-imidazol-1-yl)pyrimidine (4.0 g, 22.1 mmol)
was split into two equal portions and separately placed into two
sealed tubes with a solution of 7 N ammonia in methanol (40 mL).
The tubes were sealed and heated at 65.degree. C. for 20 h, then
allowed to stand at room temperature for 3 days. The reactions were
combined, concentrated and purified by flash chromatography on a
240 g silica gel cartridge with 40 to 100% EtOAc in hexane, then 0
to 50% MeOH in EtOAc to yield 6-(1H-imidazol-1-yl)pyrimidin-4-amine
(1.43 g, 40%). LCMS: RT=0.39 min, MH+=162.1.
Step C: Dimethyl
6-(1H-imidazol-1-yl)pyrimidin-4-ylcarbonimidodithioate
##STR01075##
[1029] To a solution of 6-(1H-imidazol-1-yl)pyrimidin-4-amine (1.0
g, 6.2 mmol) in DMF (18 mL) was added 10 M NaOH (1.24 mL, 12.4
mmol) dropwise, carbon disulfide (0.93 mL, 16 mmol), NaOH (1.24 mL,
12.4 mmol), and iodomethane (0.97 mL, 16 mmol) at 15 min intervals.
Stirring was continued for 2 h, then the mixture was poured into
water. The cloudy solution was partitioned into EtOAc, washed with
water, concentrated, and purified by flash chromatography on a 110
g silica gel cartridge with 10 to 50% EtOAc in hexane to yield
dimethyl 6-(1H-imidazol-1-yl)pyrimidin-4-ylcarbonimidodithioate
(411 mg, 25% yield). .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.79
(1H, d, J=1.01 Hz), 8.65 (1H, d, J=1.01 Hz), 7.96 (1H, t, J=1.51
Hz), 7.33 (1H, d, J=1.01 Hz), 7.12-7.19 (1H, m), 2.58 (6H, s).
LCMS: RT=0.63 min, MH+=266.1.
Step D:
(R)--N-(6-(1H-Imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-
-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01076##
[1031] A suspension of (S)-3-(aminomethyl)quinuclidin-3-ol
dihydrochloride (419 mg, 1.8 mmol), dimethyl
6-(1H-imidazol-1-yl)pyrimidin-4-ylcarbonimidodithioate (404 mg, 1.5
mmol), and cesium carbonate (1.24 g, 3.8 mmol) was stirred in DMF
(3.8 mL) at 75.degree. C. for 2 h. The reaction was concentrated
and purified by flash chromatography on a 40 g silica gel cartridge
with 0 to 3% [9:1 MeOH/NH4OH] in CHCl.sub.3 to yield
(R)--N-(6-(1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-b-
icyclo[2.2.2]octan]-2-amine (417 mg, 83% yield). .sup.1H NMR (400
MHz, CHLOROFORM-d) .delta. ppm 9.40 (1H, br. s.), 8.60 (1H, d,
J=1.01 Hz), 8.37 (1H, t, J=1.01 Hz), 7.55 (1H, t, J=1.39 Hz), 7.16
(1H, dd, J=1.39, 0.88 Hz), 6.84 (1H, br. s.), 3.98 (1H, d, J=9.57
Hz), 3.64 (1H, d, J=9.57 Hz), 3.34 (1H, dd, J=14.98, 1.64 Hz),
2.69-3.04 (5H, m), 2.08-2.21 (2H, m), 1.67-1.78 (1H, m), 1.43-1.62
(2H, m). LCMS: RT=0.26 min, MH+=326.2.
Example 348
(R)--N-(6-(4-Methyl-1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazol-
e-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01077##
[1032] Step A: 4-Chloro-6-(4-methyl-1H-imidazol-1-yl)pyrimidine
##STR01078##
[1034] A mixture of 4,6-dichloropyrimidine (11.92 g, 80 mmol),
4-methyl-1H-imidazole (6.57 g, 80 mmol), and cesium carbonate (26.1
g, 80 mmol) was stirred in DMF (50 mL) at room temperature for 18
h. The reaction was diluted into 200 mL water and extracted three
times with 200 mL EtOAc. The combined organic layers were
concentrated and purified by flash chromatography on a 300 g silica
gel cartridge with 25 to 75% ethyl acetate in hexane to yield a
mixture of the two regioisomeric products. The pooled fractions
were concentrated to about 200 mL volume and cooled to yield
4-chloro-6-(4-methyl-1H-imidazol-1-yl)pyrimidine (5.70 g, 37%
yield). The positive identification of the 4-methylimidazole
regioisomer was accomplished by its NMR NOE properties. .sup.1H NMR
(400 MHz, CHLOROFORM-d) .delta. ppm 8.80 (1H, d, J=0.50 Hz), 8.36
(1H, d, J=1.26 Hz), 7.29-7.31 (1H, m), 7.25 (1H, d, J=1.01 Hz),
2.27 (3H, d, J=1.01 Hz). LCMS: RT=0.50 min, MH+=195.1.
Step B: 6-(4-Methyl-1H-imidazol-1-yl)pyrimidin-4-amine
##STR01079##
[1036] A solution of
4-chloro-6-(4-methyl-1H-imidazol-1-yl)pyrimidine (1.18 g, 6.1 mmol)
in 2 M ammonia (20 mL, 40.0 mmol)/isopropanol was heated at
80.degree. C. in a sealed vial for 18 h. After cooling the solid
precipitate was filtered and dried to give
6-(4-methyl-1H-imidazol-1-yl)pyrimidin-4-amine (733 mg, 69.0%
yield). 1H NMR (400 MHz, DMSO-d6) .delta. ppm 8.31 (1H, d, J=1.26
Hz), 8.28 (1H, d, J=0.76 Hz), 7.49 (1 H, t, J=1.13 Hz), 7.16 (2H,
s), 6.50 (1H, d, J=0.76 Hz), 2.15 (3H, d, J=0.76 Hz). LCMS: RT=0.39
min, MH+=176.1.
Step C: Dimethyl
6-(4-methyl-1H-imidazol-1-yl)pyrimidin-4-ylcarbonimidodithioate
##STR01080##
[1038] To a solution of
6-(4-methyl-1H-imidazol-1-yl)pyrimidin-4-amine (700 mg, 4.00 mmol)
in DMF (12 mL) was added dropwise 10 M NaOH (0.8 mL, 8 mmol),
carbon disulfide (0.60 mL, 10 mmol), NaOH (0.8 mL, 8 mmol), and
iodomethane (0.62 mL, 10 mmol) at 15 min intervals. Stirring was
continued for 2 h, then the mixture was poured into water. The
cloudy solution was partitioned with EtOAc, washed with water,
concentrated, and purified by flash chromatography on a 110 g
silica gel cartridge with 50 to 100% EtOAc in hexane to yield
dimethyl
6-(4-methyl-1H-imidazol-1-yl)pyrimidin-4-ylcarbonimidodithioate
(438 mg, 39% yield). 1H NMR (400 MHz, MeOD) .delta. ppm 8.76 (1H,
d, J=1.01 Hz), 8.55 (1H, d, J=1.26 Hz), 7.62-7.66 (1H, m), 7.24
(1H, d, J=1.01 Hz), 2.57 (6H, s), 2.24 (3H, d, J=1.01 Hz). LCMS:
RT=0.66 min, MH+=280.1.
Step D:
(R)--N-(6-(4-Methyl-1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspir-
o[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01081##
[1040] A suspension of (S)-3-(aminomethyl)quinuclidin-3-ol
dihydrochloride (412 mg, 1.80 mmol), dimethyl
6-(4-methyl-1H-imidazol-1-yl)pyrimidin-4-ylcarbonimidodithioate
(419 mg, 1.5 mmol), and cesium carbonate (1.22 g, 3.75 mmol) was
stirred in DMF (3.8 mL) at 75.degree. C. for 2 h. The reaction was
concentrated and purified by flash chromatography on a 40 g silica
gel cartridge with a pre-run of 1% [95:5 MeOH/NH4OH] in EtOAc, then
1 to 2% [95:5 MeOH/NH4OH] in CHCl3 to yield
(R)--N-(6-(4-methyl-1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazo-
le-5,3'-bicyclo[2.2.2]octan]-2-amine (472 mg, 92% yield). 1H NMR
(400 MHz, CHLOROFORM-d) .delta. ppm 9.40 (1H, br. s.), 8.56 (1H, d,
J=1.01 Hz), 8.28 (1H, d, J=1.26 Hz), 7.21 (1H, s), 6.76 (1H, br.
s.), 3.97 (1H, d, J=9.57 Hz), 3.63 (1H, d, J=9.57 Hz), 3.33 (1H,
dd, J=14.86, 1.76 Hz), 2.62-3.05 (5H, m), 2.24 (3H, d, J=1.01 Hz),
2.03-2.19 (2H, m), 1.65-1.81 (1H, m), 1.39-1.62 (2H, m). LCMS:
RT=0.25, 0.46 min, MH+=340.3.
Example 349
(R)--N-(6-(4-Chloro-1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazol-
e-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01082##
[1041] Step A: 4-Chloro-6-(4-chloro-1H-imidazol-1-yl)pyrimidine
##STR01083##
[1043] A mixture of 4,6-dichloropyrimidine (1.8 g, 12 mmol),
4-chloro-1H-imidazole (1.23 g, 12.00 mmol), and cesium carbonate
(3.91 g, 12.00 mmol) was stirred in DMF (8 mL) at room temperature
for 18 h. The reaction was diluted into water and extracted with
EtOAc. The combined organic layers were concentrated and purified
by flash chromatography on a 120 g silica gel cartridge with 0 to
40% ethyl acetate in hexane to yield
4-chloro-6-(4-chloro-1H-imidazol-1-yl)pyrimidine (1.41 g, 54.6%
yield). The positive identification of the 4-chloroimidazole
regioisomer was accomplished by its NMR NOE properties. 1H NMR (400
MHz, CHLOROFORM-d) .delta. ppm 8.86 (1H, d, J=0.76 Hz), 8.33 (1H,
d, J=1.51 Hz), 7.54 (1H, d, J=1.76 Hz), 7.30 (1H, d, J=0.76 Hz).
LCMS: RT=0.81 min, MH+=215.1
Step B: 6-(4-Chloro-1H-imidazol-1-yl)pyrimidin-4-amine
##STR01084##
[1045] A solution of
4-chloro-6-(4-chloro-1H-imidazol-1-yl)pyrimidine (0.99 g, 4.60
mmol) in 2 M ammonia (20 mL, 40.0 mmol)/isopropanol was heated at
80.degree. C. in a sealed vial for 18 h. After cooling the solid
precipitate was filtered and dried to give
6-(4-chloro-1H-imidazol-1-yl)pyrimidin-4-amine (884 mg, 98% yield).
1H NMR (400 MHz, DMSO-d6) .delta. ppm 8.44 (1H, d, J=1.51 Hz), 8.32
(1H, d, J=0.76 Hz), 7.96 (1 H, d, J=1.51 Hz), 7.30 (2H, br. s.),
6.58 (1H, d, J=1.01 Hz). LCMS: RT=0.57 min, MH+=196.1.
Step C: Dimethyl
6-(4-chloro-1H-imidazol-1-yl)pyrimidin-4-ylcarbonimidodithioate
##STR01085##
[1047] To a solution of
6-(4-chloro-1H-imidazol-1-yl)pyrimidin-4-amine (850 mg, 4.35 mmol)
in DMF (12 mL) was added dropwise 10 M NaOH (0.87 mL, 8.7 mmol),
carbon disulfide (0.65 mL, 10.9 mmol), NaOH (0.87 mL, 8.7 mmol),
and iodomethane (0.68 mL, 10.9 mmol) at 15 min intervals. Stirring
was continued for 2 h and the mixture was poured into water. The
cloudy solution was partitioned into EtOAc, washed with water,
concentrated, and purified by flash chromatography on a 110 g
silica gel cartridge with 10 to 40% EtOAc in hexane to yield
dimethyl
6-(4-chloro-1H-imidazol-1-yl)pyrimidin-4-ylcarbonimidodithioate
(558 mg, 43% yield). 1H NMR approx. 9:1 mixture of rotamers (400
MHz, MeOD) .delta. ppm 8.80 (0.8H, d, J=0.76 Hz), 8.79 (0.2H, d,
J=0.50 Hz), 8.59 (1H, d, J=1.51 Hz), 8.50 (0.1H, d, J=1.26 Hz),
7.97 (0.9H, d, J=1.51 Hz), 7.84 (0.1H, d, J=1.51 Hz), 7.32 (0.9H,
d, J=0.76 Hz), 2.63 (0.6H, s), 2.58 (5.4H, s). LCMS: RT=0.94 min,
MH+=300.0
Step D:
(R)--N-(6-(4-Chloro-1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspir-
o[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01086##
[1049] A suspension of (S)-3-(aminomethyl)quinuclidin-3-ol
dihydrochloride (412 mg, 1.800 mmol), dimethyl
6-(4-chloro-1H-imidazol-1-yl)pyrimidin-4-ylcarbonimidodithioate
(450 mg, 1.5 mmol), and cesium carbonate (1.22 g, 3.75 mmol) was
stirred in DMF (3.8 mL) at 75.degree. C. for 2 h. The reaction was
concentrated and purified by flash chromatography on a 40 g silica
gel cartridge with a pre-run of 1% [95:5 MeOH/NH4OH] in EtOAc, then
1 to 2% [95:5 MeOH/NH4OH] in CHCl3 to yield
(R)--N-(6-(4-chloro-1H-imidazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazo-
le-5,3'-bicyclo[2.2.2]octan]-2-amine (265 mg, 48% yield). 1H NMR
(400 MHz, CHLOROFORM-d) .delta. ppm 9.41 (1H, br. s.), 8.60 (1H, d,
J=1.01 Hz), 8.25 (1H, d, J=1.51 Hz), 7.44 (1H, d, J=1.51 Hz), 6.78
(1H, s), 4.00 (1H, d, J=9.82 Hz), 3.67 (1H, d, J=9.57 Hz), 3.36
(1H, dd, J=14.86, 1.51 Hz), 2.73-3.06 (5H, m), 2.09-2.22 (2H, m),
1.69-1.79 (1H, m), 1.46-1.64 (2H, m). LCMS: RT=0.54 min,
MH+=360.2.
Example 350
(R)--N-(6-(1H-Pyrazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bic-
yclo[2.2.2]octan]-2-amine
##STR01087##
[1050] Step A: 4-Chloro-6-(1H-pyrazol-1-yl)pyrimidine
##STR01088##
[1052] A mixture of 4,6-dichloropyrimidine (5.96 g, 40 mmol),
1H-pyrazole (2.72 g, 40.0 mmol), and cesium carbonate (13.03 g,
40.0 mmol) was stirred in DMF (25 mL) at room temperature for 18 h.
The reaction was diluted into 100 mL water and extracted with
EtOAc. The combined organic layers were concentrated and purified
by flash chromatography on a 240 g silica gel cartridge with 0 to
20% ethyl acetate in hexane to yield
4-chloro-6-(1H-pyrazol-1-yl)pyrimidine (4.80 g, 66% yield). 1H NMR
(400 MHz, CHLOROFORM-d) .delta. ppm 8.77 (1H, d, J=0.76 Hz), 8.55
(1H, d, J=2.77 Hz), 7.96 (1H, d, J=1.01 Hz), 7.80 (1H, d, J=1.01
Hz), 6.51 (1H, dd, J=2.64, 1.64 Hz).
[1053] LCMS: RT=0.86 min, MH+=181.1
Step B: 6-(1H-Pyrazol-1-yl)pyrimidin-4-amine
##STR01089##
[1055] A solution of 4-chloro-6-(1H-pyrazol-1-yl)pyrimidine (1.31
g, 7.25 mmol) in 2 M ammonia (20 mL, 40.0 mmol)/isopropanol was
heated at 80.degree. C. in a sealed vial for 24 h. The reaction was
stored at room temperature for 4 days. The solid precipitate was
filtered and dried to give 6-(1H-pyrazol-1-yl)pyrimidin-4-amine
(1.15 g, 98% yield). 1H NMR (400 MHz, DMSO-d6) .delta. ppm 8.53
(1H, dd, J=2.52, 0.50 Hz), 8.30 (1H, d, J=0.76 Hz), 7.82 (1H, d,
J=1.01 Hz), 7.19 (2H, br. s.), 6.88 (1H, d, J=1.01 Hz), 6.55 (1H,
dd, J=2.64, 1.64 Hz). LCMS: RT=0.52 min, MH+=162.1.
Step C: Dimethyl
6-(1H-pyrazol-1-yl)pyrimidin-4-ylcarbonimidodithioate
##STR01090##
[1057] To a solution of 6-(1H-pyrazol-1-yl)pyrimidin-4-amine (1.0
g, 6.20 mmol) in DMF (18 mL) was added dropwise 10 M NaOH (1.24 mL,
12.4 mmol), carbon disulfide (0.933 mL, 15.5 mmol), NaOH (1.24 mL,
12.4 mmol), and iodomethane (0.966 mL, 15.5 mmol) at 15 min
intervals. Stirring was continued overnight and the mixture was
poured into water. The tan precipitate was filtered, washed with
water, and dried to yield dimethyl
6-(1H-pyrazol-1-yl)pyrimidin-4-ylcarbonimidodithioate (136 mg, 8%
yield).
[1058] 1H NMR (400 MHz, MeOD) .delta. ppm 8.74 (1H, d, J=1.01 Hz),
8.64 (1H, d, J=2.52 Hz), 7.82 (1H, d, J=1.26 Hz), 7.40 (1H, d,
J=1.01 Hz), 6.57 (1H, dd, J=2.64, 1.64 Hz), 2.58 (6H, s). LCMS:
RT=0.96 min, MH+=266.1.
Step D:
(R)--N-(6-(1H-Pyrazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole--
5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01091##
[1060] A suspension of (S)-3-(aminomethyl)quinuclidin-3-ol
dihydrochloride (135 mg, 0.59 mmol), dimethyl
6-(1H-pyrazol-1-yl)pyrimidin-4-ylcarbonimidodithioate (130 mg, 0.49
mmol), and cesium carbonate (399 mg, 1.23 mmol) was stirred in DMF
(1.2 mL) at 75.degree. C. for 2 h. The reaction was concentrated
and purified by flash chromatography on a 40 g silica gel cartridge
with a pre-run of 1% [95:5 MeOH/NH4OH] in EtOAc, then isocratic 1%
[95:5 MeOH/NH4OH] in CHCl3 to yield
(R)--N-(6-(1H-pyrazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bi-
cyclo[2.2.2]octan]-2-amine (105 mg, 66% yield). 1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 9.46 (1H, br. s.), 8.58 (1H, d, J=1.01
Hz), 8.50 (1H, dd, J=2.64, 0.63 Hz), 7.74 (1H, d, J=1.01 Hz), 7.47
(1H, br. s.), 6.44 (1H, dd, J=2.64, 1.64 Hz), 3.98 (1H, d, J=9.57
Hz), 3.65 (1H, d, J=9.57 Hz), 3.39 (1H, dd, J=14.86, 1.51 Hz),
2.71-3.09 (5H, m), 2.10-2.26 (2H, m), 1.69-1.80 (1H, m), 1.48-1.64
(2H, m). LCMS: RT=0.51 min, MH+=326.2.
Example 351
(R)--N-(6-(1H-1,2,4-triazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,-
3'-bicyclo[2.2.2]octan]-2-amine
##STR01092##
[1061] Step A: 4-Chloro-6-(1H-1,2,4-triazol-1-yl)pyrimidine
##STR01093##
[1063] A mixture of 4,6-dichloropyrimidine (5.96 g, 40 mmol),
1H-1,2,4-triazole (1.381 g, 20.00 mmol), and cesium carbonate
(13.03 g, 40.0 mmol) was stirred in DMF (25 mL) at room temperature
for 16 h. The reaction was diluted with 100 mL water and extracted
with EtOAc. The combined organic extracts were concentrated and
purified by flash chromatography on a 160 g silica gel cartridge
with 25 to 50% EtOAc in hexane to yield
4-chloro-6-(1H-1,2,4-triazol-1-yl)pyrimidine (2.78 g, 38% yield).
1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 9.21 (1H, s), 8.87 (1H,
s), 8.15 (1H, s), 7.91 (1H, s). LCMS: RT=0.68 min, MH+=182.1.
Step B: 6-(1H-1,2,4-Triazol-1-yl)pyrimidin-4-amine
##STR01094##
[1065] A solution of 4-chloro-6-(1H-1,2,4-triazol-1-yl)pyrimidine
(1.24 g, 6.83 mmol) in 2 M ammonia (20 mL, 40.0 mmol)/isopropanol
was heated at 80.degree. C. in a sealed vial for 18 h. After
cooling the solid precipitate was filtered and dried to give
6-(1H-1,2,4-triazol-1-yl)pyrimidin-4-amine (1.06 g, 96% yield). 1H
NMR (400 MHz, DMSO-d6) .delta. ppm 9.30 (1H, s), 8.35 (1H, s), 8.30
(1H, s), 7.39 (2H, br. s.), 6.83 (1 H, d, J=0.76 Hz). LCMS: RT=0.41
min, MH+=163.1
Step C: Dimethyl
6-(1H-1,2,4-triazol-1-yl)pyrimidin-4-ylcarbonimidodithioate
##STR01095##
[1067] To a solution of 6-(1H-1,2,4-triazol-1-yl)pyrimidin-4-amine
(1.00 g, 6.17 mmol) in DMF (18 mL) was added dropwise 10 M NaOH
(1.23 mL, 12.3 mmol), carbon disulfide (0.927 mL, 15.4 mmol), NaOH
(1.23 mL, 12.3 mmol), and iodomethane (0.96 mL, 15 mmol) at 15 min
intervals. Stirring was continued for 2 h and the mixture was
poured into water. The cloudy solution was partitioned with EtOAc,
washed with water, concentrated, and purified by flash
chromatography on a 110 g silica gel cartridge with 10 to 50% EtOAc
in hexane to yield dimethyl
6-(1H-1,2,4-triazol-1-yl)pyrimidin-4-ylcarbonimidodithioate (541
mg, 33% yield). 1H NMR 4:1 mixture of rotamers (400 MHz, MeOD)
.delta. ppm 9.42 (0.74H, s), 9.40 (0.35H, s), 9.04 (0.32H, s), 8.84
(0.76H, d, J=1.01 Hz), 8.81 (0.33H, d, J=1.01 Hz), 8.23 (1H, s),
7.42 (0.75H, d, J=1.01 Hz), 2.64 (1.2H, s), 2.59 (4.8H, s). LCMS:
RT=0.86 min, MH+=267.1.
Step D:
(R)--N-(6-(1H-1,2,4-Triazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[ox-
azole-5,3'-bicyclo[2.2.2]octan]-2-amine
##STR01096##
[1069] A suspension of (S)-3-(aminomethyl)quinuclidin-3-ol
dihydrochloride (412 mg, 1.800 mmol), dimethyl
6-(1H-1,2,4-triazol-1-yl)pyrimidin-4-ylcarbonimidodithioate (400
mg, 1.5 mmol), and cesium carbonate (1222 mg, 3.75 mmol) was
stirred in DMF (3.8 mL) at 75.degree. C. for 2 h. The reaction was
concentrated and purified by flash chromatography on a 40 g silica
gel cartridge with a pre-run of 1% [95:5 MeOH/NH4OH] in EtOAc, then
1 to 3% [95:5 MeOH/NH4OH] in CHCl3 to yield
(R)--N-(6-(1H-1,2,4-triazol-1-yl)pyrimidin-4-yl)-4H-1'-azaspiro[oxa-
zole-5,3'-bicyclo[2.2.2]octan]-2-amine (210 mg, 43% yield). 1H NMR
(400 MHz, CHLOROFORM-d) .delta. ppm 9.45 (1H, br. s.), 9.14 (1H,
s), 8.61 (1H, d, J=1.01 Hz), 8.08 (1H, s), 7.39 (1H, s), 4.00 (1H,
d, J=9.57 Hz), 3.67 (1H, d, J=9.57 Hz), 3.38 (1H, dd, J=14.86, 1.76
Hz), 2.71-3.08 (5H, m), 2.08-2.23 (2H, m), 1.68-1.80 (1 H, m),
1.45-1.64 (2H, m). LCMS: RT=0.46 min, MH+=327.2
Example 352
(S)--N-(Isoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-
-2-amine
##STR01097##
[1071]
(S)--N-(Isoquinolin-3-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2-
]octan]-2-amine was prepared by the method of Example 239 (steps
A-B), starting from (R)-3-(aminomethyl)quinuclidin-3-ol
dihydrochloride. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.00 (1H,
s), 7.91 (1H, d, J=8.06 Hz), 7.70 (1H, d, J=8.31 Hz), 7.58 (1H, t,
J=7.18 Hz), 7.40 (1H, t, J=7.05 Hz), 7.30 (1H, br. s.), 3.95 (1H,
d, J=9.82 Hz), 3.64 (1H, d, J=10.07 Hz), 3.16-3.25 (1H, m),
3.02-3.13 (1H, m), 2.92 (2H, t, J=7.30 Hz), 2.80 (2H, t, J=7.05
Hz), 2.05-2.23 (2H, m), 1.52-1.84 (3H, m). MS (LC/MS) R.T.=1.39;
[M+H].sup.+=309.21.
[1072] It will be evident to one skilled in the art that the
present disclosure is not limited to the foregoing illustrative
examples, and that it can be embodied in other specific forms
without departing from the essential attributes thereof. It is
therefore desired that the examples be considered in all respects
as illustrative and not restrictive, reference being made to the
appended claims, rather than to the foregoing examples, and all
changes which come within the meaning and range of equivalency of
the claims are therefore intended to be embraced therein.
* * * * *